proceedings of the 24th autumn meeting from october 14–16, 1982-yamagata city, japan

GastroenterologiaJaponica Vol. 18, No. 2 Copyright �9 1983 by The Japanese Society of Gastroenterology Printed in Japan

P R O C E E D I N G S O F T H E 2 4 T H A U T U M N M E E T I N G

From October 14-16 , 1 9 8 2 m Y a m a g a t a Ci ty , Japan

C h a i r m a n : Dr. M a k o t o I S H I K A W A , M . D .

C o - M o d e r a t o r s :

S y m p o s i u m (1):

Gastric Cancer

Dr. T a k u K A W A C H I and Dr. T a k e o N A G A Y O

1. Experimental gastric carcinoma in rat as an animal model

Michihito TAKAHASHI

Dept Pathol, Natl lnst Hygfen Scf, Tokyo

Experimental animal model of gastric carcinoma

is available using rats administered MNNG. A num-

ber of investigations have now done as the most re-

liable tool for studying pathogenesis and histogene-

sis of gastric cancer.

1. Similarity of experimental gastric carcinoma

to human gastric carcinoma.

The model closely resembles the human gastric

carcinoma both by macroscopical and microscopi-

cal appearances.

2. Biological behavior of experimental gastric

carcinoma.

Certain types of well-differentiated adenocarcino-

mas show low malignancy but poorly differentiated

adenocarcinomas clearly show malignant nature.

3. Effect of ulcer on experimental gastric car-

cinogenesis.

We used this model to study the pathogenesis of

ulcer-cancer. Oral administration of iodoacetamide

in drinking water produced ulcer in the fundic re-

gion of the stomach. MNNG administration after

this treatment produced adenocarcinoma in the

region where the ulcer formed. This phenomenon

has been confirmed by freezing ulcer method.

4. Usefulness of experimental gastric carcino-

mas in rats.

This model is very useful. Because rats are more

economic and can be kept enough numbers for

statistical evaluation. We are able to obtain reliable

results in repeated experiments and easily to keep

constant condition.

2. Analytical study of rat stomach carcinogenesis induced by M N N G

Oichiro KOBORI

1st Dept Surg, Fac Mecl, Unfv of Tokyo, Tokyo

Thirty-seven young male Wistar rats were used for

this experiment. Twenty-seven rats were given

MNNG solution at a concentration of 80/~g/ml for

13 weeks. Group 1 (12 rats) were given normal tap

water, observed for 55 weeks and sacrificed. Group 2

(15 rats) had choledochogastrostomy on the 20th

week after the stop of the administration of MNNG.

At the operation, choledochus was severed at the

midpoint and the proximal cut-end was anasto-

mosed to the forstomach using a fine polyethylene

tube. The distal cut-end was ligated. In Group 2, 10

rats could live longer than 10 weeks as the effective

number of rats and all of them were sacrificed be-

fore the 33rd week. Average survival t ime of these

rats was 17 weeks after the operation. Group 3 (10

rats) had the same operation without previous ad-

ministration of MNNG, 6 rats of which could live

longer than 10 weeks and were sacrificed as the con-

trol group from the 1 l th to the 33rd week.

The stomach mucosa of Group 1 rats showed

slight focal hyperplastic gastritis. In Group 2, re-

markable hyperplastic gastritis was observed.

Another remarkable histological change was cystic

glandular dilatation extensively observed in the

whole stomach mucosa of Group 2 and 3 rats. Two

stomach carcinomas were found only in Group 2

rats.

Hyperplastic gastritis is usually found in the

April 1983 Proceedings of 24th Autumn Meeting 149

human stomach mucosa and the cystic glandular

dilatation was compatible with the characteristic

histological features documented in human residual

stomach mucosa exposed to the bile regurgitation.

We wanted to stress that the background lesions of

human stomach carcinogenesis were experimentally

induced by the lower doses of the carcinogen, longer

period of observation, and the addition of the possi-

ble cocarcinogenic substance by a rout mimicking

physiological conditions.

3, Comparative studies on the carcinoma of the gastric remnant in rats

and humans

Nobuaki KAIBARA and Hideaki NISHIDOI

1st Dept Surg, Tottori Univ Sch Med, Yonago

Gastric carcinomas were induced with N-methyl-

N'-nitro-N-nitrosoguanidine (MNNG) in rats that

had undergone partial gastrectomy removing half of

the glandular stomach. Carcinomas developed in

the remnant stomach with the incidence being

38.9% in rats that had received Billroth II recon-

struction, 25.0% in those with short Roux-Y recon-

struction (10cm distance between gastrojejunosto-

mia and jejunojejunostomia), 7 .1% in those with

long Roux-Y reconstruction (30cm distance be-

tween the 2 anastomoses) and 9.5 % in the controls,

indicating higher incidence of carcinoma of the gas-

tric remnant in rats with greater amount of

duodeno-gastric reflux. These results suggest that

the reflux of duodenal juice (including bile) is sub-

stantially related to the development of MNNG-in-

duced remnant gastric carcinoma.

In our series of patients with carcinoma of the gas-

tric remnant, site of the tumors was on the stoma in

9 of 11 cases, which was in accordance with the ani-

mal model. Polypoid lesion, termed histologically as

gastritis cystica polyposa, has been interpreted as

being related to the development of cancer. In the

present study, a case of gastritis cystica polyposa

with intramucosal carcinoma was presented.

4. Experimental gastric cancer as animal model of human gastric cancer

by N-ethyl-N'-nitro-N- nitrosoguanidine

Norio MATSUKURA and Akiro SHIROTA

1st Dept Surg, Nippon Med Sch, Tokyo

1) To establish the method of inducing gastric

cancer in dogs as an animal model of human

gastric cancer, N-ethyt-N'-nitro-N-nitrosoguanidine

(ENNG) was given to 15 male beagle dogs at a con-

centration of 100 or 150 ug /ml as drinking water. In

six dogs given 100 ug E N N G / m l for 6-9 months, de-

pressed type of early gastric cancer and Borrman 2

or 3 types of advanced gastric cancer were induced.

On the contrary, to 9 dogs given 150 ug E N N G / m l

elevated and depressed type of early gastric cancer,

Borrman 3 type and unclassified type advanced gas-

tric cancer were induced. Six to 9 months are suit-

able period for administration of 100 ug E N N G / m l

to induced gastric cancer in dogs as animal model of

human gastric cancer.

2) Labelling index (LI) of canine gastric cancer

induced by ENNG was examined in vivo by pulse in-

jection of [methyl -s] thymidine intravenously. LIs of

cancerous lesions were lower than those of non-can-

cerous gastric mucosa. The mean value of LIs of

well-differentiated type of adenocarcinoma in the

mucosa was higher than that of poorly differentiated

type. In the well-differentiated type of adenocar-

cinoma, the value of LI (y) decreased sharply with

the increase in the diameter (x) of the carcinoma

(y=6.645/x ~ p<0.05). On the contrary, the

value of LI (y) increased proportionally with the in-

crease in the diameter (x) of carcinoma in the poorly

differentiated type (y=0.213x + 0.95; y=0.535;

p<0.05).

5. Experimental gastric cancer of dogs

from a view poin t of follow-up studies of gastric carcinogenesis

Takao SAITO

1st Dept Surg, Med Coil Oita, Oita

Experimental gastric cancer of dogs was periodi-

cally observed during its entire course by endoscopy

150 Proceedings of 24th Autumn Meeting Vol. 18, No. 2

and biopsy and following results were obtained.

1. As preceding lesions of experimental gastric

cancer, superficial erosion, deep erosion, ulcer,

healed erosion, healed ulcer and hyperplastic polyp

were observed.

2. Atypical glands, a border line lesion which

was observed during the course of gastric carcino-

genesis, were classified into two types according to

histogram-patterns of nuclear DNA contents, i.e.,

regenerative type and cancer type. Atypical glands

which showed the latter type were considered as a

precancerous lesion.

3. From studies of micro-cancer in experimental

gastric cancer, it is presumed that a well differen-

tiated carcinoma developed in severely eroded

mucosa while a poorly differentiated carcinoma de-

veloped in slightly or not eroded mucosa.

4. Experimental gastric cancer showed several

patterns in the growth of early cancer to advanced

cancer. Malignant cycle was observed during the

stage of early cancer.

6. Histogenetic and clinical significances of the experimental canine

gastric cancer

Masakatsu SUNAGAWA and Kazuo HOSHI

1st Dept Surg, Tokyo Med and Dent Univ, Sch Med., Tokyo

Sixty-six adult beagles were given N-ethyl-N'-

nitro-N-nitrosoguanidine (ENNG) by our method of

administration called "milk method". The develop-

ment of carcinoma and the effect of anticancer

agent on the experimental cancer were studied and

the following results were obtained.

1. Signet-ring cell and poorly differentiated

adenocarcinomas were induced by three months ad-

ministration of ENNG. These cancer cells appeared

to develop from the peculiar "double layer struc-

ture" of the foveolar epithelium. Well differentiated

adenocarcirmma was induced by more than six

months administration. Minute loci of the carcino-

ma were found in normal epithelium, at the bot tom

of hyperplastic epithelium and at top of hyperplastic

polyp.

2. Fourteen beagles were operated by the same

procedure for human gastric cancer and followed-

up by means of endoscopy and biopsy. The polipoid

and ulcerative lesions were found in the gastric rem-

nant six months after the operation. The lesions

were showed to be carcinoma after eight months to

four years period, The carcinomas were considered

to be heterochronous multiple cancers.

3. There was no significant difference in 5-Fu

concentrations in normal tissures, cancerous tissures

and lymph nodes compared with those of human

gastric cancer. Fu-O-G, a derivative of 5-Fu and M-

83, a derivative of MMC, could be given to the

beagles of gastric cancer for two or three months

without intolerable toxic reactions. Certain changes

of gross findings of the carcinoma were observed by

endoscopy in two of ten dogs treated by Fu-O-G or

M-83.

7. Application of dogs with ENNG- induced gastric carcinoma to

chemotherapeutic study

Minoru KURIHARA

Dept Intern Med, Toyosu Hosp, Showa Univ, Tokyo

Tsuguhiko IZUMI

Dept Intern Med (Gastroenterol), Sch Med, Juntendo Univ, Tokyo

Gastric adenoearcinoma is prepared in dogs ex-

perimentally by oral administration of N-ethyl-N'-

nitro-N-nitroso-guanidine (ENNG) in the form of

wet pellet. We reported first this procedure just ten

years ago in this Congress. Our method has been

come into wide use, because it has provided us with

excellent models for the following three reasons: 1)

Using our method the target organ is l imited only to

the stomach. 2) This method can induce advanced

gastric adenocarcinoma with metastasis to distant

lymph nodes, liver, lungs, bones and skin. 3)

ENNG-induced dog carcinoma cells can be trans-

planted to nude mice. Furthermore, combination of

ENNG and subcutaneous injection of gastrin could

develop scirrhous carcinoma which resulted in pylo-

ric stenosis in three out of the ten beagles. UFT was

orally administered to nine dogs with gastric car-

cinoma induced by ENNG in order to evaluate the


effectiveness of UFT. Three animals were allocated

to 3 groups each. Daily dose and duration of ad-

ministration were as follows: 5 m g / k g for 101 days

in Group I, 7.5 mg /kg for 67 days in Group II, and

12 m g / k g for 45 days in Group III respectively. X-

ray and endoscopic findings revealed very few

changes in the dogs. Grade IIB, i.e. clear cytostatic

effects with few survived tumor cells, was available

in Group III on the basis of histological findings

(Ohoshi's histological criteria of chemotherapeutic

effects). 5-FU levels in autopsied organs were mea-

sured four hours after UFT administration to these

animals. They were high in tumor tissues as com-

pared with adjacent normal tissues in the stomach.

8. A model for the evaluation of anti- cancer agents by an application of

canine gastric carcinoma

Gentsu UOZUMI and Yuzo AKASAKA

Dept Prevent Med, Kyoto Prefect Univ Med, Kyoto

A model for the evaluation of anti-cancer agents

was established by an application of ENNG induced

canine gastric carcinoma which has been reported as

a model of growth and development of human gas-

tric carcinoma. Thirty one beagles were given 75 mg

ENNG a day for eight months. OK-432 and Glucan,

Schizophyllan (SPG) were used as anti-cancer agents

in this experiment. The beagles were divided into

five groups. (control. different doses and different

terms of SPG. OK-432) These were injected intra-

muscularly twice a week. All beagles were followed

up by endoscopy and biopsy for at most 36 months

and autopsy was carried out in each dogs. By four-

teen months adenocarcinoma and signet-ring cell

carcinoma of the stomach was revealed in all

beagles. Survival rate of all dogs at the 36th month

was 22.6% (7/31). By Generalized Wilcoxon test,

there is a statistically significant difference in sur-

vival curves between control group and early ther-

apeutic group. On the other hand, multivariate

analysis revealed differences of survival term of each

family of tumor bearing beagle. This model is useful

of the evaluation of anti-cancer agents because of its

several advantages that the tumor is autochthonus

and able to be followed endoscopically as the same

way in human gastric carcinoma.


B l o o d C i r c u l a t i o n and Gastroenterologica l Diseases

C o - M o d e r a t o r s : Dr. Fusah i ro N A G A O a n d Dr. M a s a h a r u T S U C H I Y A

1. Relationship between mucosal blood flow

and the development of acute gastric mucosal lesion

Norio NAKAMURA and Fusahiro NAGAO

2nd Dept Surg, Jikei Univ Sch Med, Tokyo

Concerning the etiology of acute gastric mucosal

lesion, an abrupt circulatory impediment in the gas-

tric mucosal membrane is concidered to be the

direct causal factor for this disease. In this study, the

mechanism of circulatory impediment after stress

binding was discussed, dividing it into the gastric,

l~Oric and duodenal gland area.

Materials and Methods: 1) As an experimental

m~mal , male wistar-strain rats were used. The

rumination of acute gastric mucosal lesion was per-

formed according to the water-immersion restraint

method. 2) For the measurement of mucosal blood

flow, the tissue blood flow meter of hydrogen-gas

clearance type was used. 3) Silicon rubber was in-

fused and the blood vessel casts of the capillary con-

struction of rat stomach and duodenum were ob-

served under stereoscopic microscope. 4) Tissue

catecholamines were measured by high performance

liquid chromatography with electro-chemical de-

tecter method.

Results: 1) After three hours' stress a sharp de-

crease of blood flow in the fundic gland area was ob-

served and acute gastric mucosal lesion began to de-

velop on the same area. According to the degree of

drop of mucosal blood flow, the lesion became more

extended and enlarged. 2) A difference in the


mucosal capillary construction among the fundic,

pyloric and duodenal gland area was clearly de-

tected by means of silicon rubber infusion. In the

gastric fundic gland area, the mucosal capillary was

fine and easy to develop the circulatory disturbance

by stress loading. After stress loading, the changes of

the construction were most remarkable especially in

the fundic gland area. 3) The content of tissue cate-

cholarnines was different in each portion. In the fun-

dic gland area the large quantities of catechola-

mines were consumed after stress loading.

Conclusion: As a reasonable causes of the sharp

decrease of blood flow in the fundic gland area, the

special capillary construction and the release of tis-

sue catecholamines of this area were considered to

be most important factor.

2. Relationship between appearance of

peri-nuclear vacuoles of gastric arterial media and autonomic

nervous system

Kazuro I T O H and Yutaka MATSUO

3rd Dept Intern Mecl, Nihon Univ Sch Med, Tokyo

The peri-nuclear vacuole (PNV) of arterial

smooth muscle cell in the kidney was pointed out to

be a morphological significance of vasospasm by

Tadashi Takeuchi in 1974.

The number of PNV is very small under the phys-

iological conditions but increase sharply after re-

straint and immersion in water.

The rates of increase were particularly high in the

submucosal layer.

The PNV is essentially considered arising from in-

trusion regarded as a structural characteristic of the

smooth muscle cell.

The occurrence rate of PNV is directly propor-

tinal to calculated constriction rate but arterial

caliber measured by preparation of casting speci-

men and local blood flow are inversely proportional.

Appearance of PNV in restraint and immersion

in water is not inhibited by truncal vagotomy but it

is markedly inhibited by sympathectomy.

Appearance of PNV is the morphological index of

vasospasm substantiated by the calculated constric-

tion rate, measurement of the arterial caliber by

preparation of a casting specimen and local blood

flOW.

3. The gastric submucosal blood flow of unanesthetized and water immersed-

restrained rats--Particularly the effects of the autonomic nerve

and catecholamines

Yozo W A T A N A B E and Takeshi KUDO

1st Dept Surg, Juntendo Univ, Sch Med, Tokyo

Aim: Using the hydrogen clearance technique,

this study was designed to measure the gastric sub-

mucosal blood flow of unanesthetized and restrained

water immersed rats. Furthermore, we studied the

effects of the autonomic nerve, catecholamines and

adrenal glands on the gastric blood flow when sub-

jected to restraint-water immersion stress of rats.

Methods: The subjects were male Wistar strain

rats weighing 180 200g. The blood flow was mea-

sured by hydrogen clearance technique. The wire

type plat inum electrodes were implanted into the

submucosa of oxyntic and pyloric gland area. Stress

was applied to the rats by restraint-water immersion

according to the method of Okabe and Takagi. The

experimental sample was composed of the following

12 groups. 1. Control group, 2. Surgical vagotomy

(S.V.) group, 3. Medical Vagotomy (M.V.) group,

4. Surgical sympathectomy (S.Sp.) group, 5. Medi-

cal sympathectomy (M.Sp.) group, 6. Epinephrine

(Ep.) treated group, 7. Norepinephrine (NE.)

treated group, 8. Dopamine (DA.) treated group, 9.

Adrenallectomy (Ad.) group, 10. Ad. + Ep. group,

11. AD. + N E . group and 12. Ad. + D A . group.

Results and Conclusion: 1) In control group, the

gastric submucosal blood flow showed an approxi-

mately 50% decrease, 30 minutes after subjection to

stress, and this trend continued for 4 hours. 2) In the

S.V. and M.V. group, the blood flow increased after

vagotomy, but showed a slight decrease after stress

application. The ulcer index was decreased. 3) In

the S.Sp. group, the decrease in blood flow after

stress loading was marked, and this condition per-

sisted for a long time. The ulcer index was also sig-

nificantly increased. 4) In the Ad. group the reduc-


tion in blood flow was increased and no subsequent

increase was noted. The ulcer index increased sig-

nificantly, but when catecholamines, particularly

epinephrine, are administered the decrease in blood

flow is inhibited.

Addi t iona l comment I) Pa thophys io log i ca l

studies on the format ion o f acute and

chronic gastric u lcer w i th special

reference to microc ircu la t ion

M. KITAJIMA and S. SOHMA

1st Dept Surg, Sch Med, Kyorin Univ, Tokyo

We have investigated the pathophysiological fac-

tors relating to the formation of acute and chronic

gastric ulcers and pharmacological effects of anti-

ulcerogenic agents with special reference to micro-

circulation. Most of attentions have been recently

directed toward the defensive mechanism of gastric

mucosa in comparison with offensive mechanism 1,2)

as a cause of ulcer formation. Because destruction of

defensive factors likely renders it more susceptible to

damage by hydrogen ions and pepsin. The evidence

abovementioned was speculated in acute and

chronic gastric ulcers on the basis of our clinical ex-

periences. Experimental studies which were divided

into two categories, such as, 1) stress ulcer in rats

with 30% burn, 2) chronic gastric ulcer in dog in-

fused with 10% formalin into left gastric artery,

were performed to evaluate the evidence. The

mucosal blood flow, mucosal energy metabolism,

tissue 02 saturation and microvascular structure

were studied as defensive factors. On the other

har~d, acid and pepsin output were also measured as

offensive factors. Especially, microvascular structure

of the stomach between dog infused 10% formalin

and clinical patients with chronic gastric ulcer was

compared from the standpoint of incurable and re-

current problems. Experimental results showed that

defensive factors were significantly decreased and

weakened in both types of gastric ulcer, and acid

output was not also increased in stressful state as ex-

pected. In conclusion, it appeared that the impair-

ment of gastric microcirculation might have been

intimately related to the formation of acute and

chronic gastric ulcer.

References: 1) Kitajima M, et al: J Trauma 18:

644-650, 1978. 2) Kitajima M, et al: Gastroen-

terologiaJpn 13: 175-183, 1978

Addi t iona l c o m m e n t 2) T h e in f luence of

vagotomy a n d H2 receptor antagonist on the mucosa l b lood flow of

e x p e r i m e n t a l stress

u lcer in rats

Mitsugi SUGIYAMA and Shuji TSUCHIYA

2rid Dept Surg, Yokohama City Univ Sch Med, Yokohama

The etiology of acute gastric mucosal lesions such

as stress induced ulcer or AGML is unknown, conse-

quently the therapy is not established and the clini-

cal result is unsatisfactory. In order to study the in-

fluence of vagotomy and H2 receptor antagonist on

the mucosal blood flow of experimental stress ulcer

in rats, the following three points were studied.

1. The change of microcirculation in the

stomach after loading stress was investigated mor-

phologically.

2. The change of the blood flow in the walls of

stomach and duodenum was measured with the

lapse of time by ssSr. microsphere method and

hydrogen gas clearance method.

3. The effect of total truncal vagotomy and H2

receptor antagonist.

The mucosal blood flow under the ether anesthe-

sia was 52.4 _+ 3.2 m l / m i n / 1 0 0 g in the corpus of

stomach, 38.1 _+ 3.3 m l / m i n / 1 0 0 g in the antrum,

and 104.3 + 5.0 m l / m i n / 1 0 0 g in the duodenum.

These values were significantly low and obviously in-

dicated mucosal ischemia with the lapse of after

burn. In the total truncal vagotomy and H~ receptor

antagonist treated groups, the reduction of the

mucosal blood flow after the load of stress was less

than that in the non-treated burn group. These

treatments can be expected to prevent experimental

stress induced ulcer from the aspect of improvement

of the mucosal blood flow, as well as from the aspect

of the main purpose of acid reduction.

References: 1) Delancy Jp: Surgery 84:190-193,

1978. 2) Semb BKH: Scand J Gastroente 14: 641,

1979. 3) Sugiyama M, et al: The Jpn J Gastroen-

154 Proceedzhgs of 24th Autumn Meeting Vol. 18, No. 2

terology Surgery 12(7): 417-424, 1979. 4) William

A: Surgery 3: 248, 1978.5) O'NeilJA: Surg 67: 267, 1970

tric mucosal blood vessels and blood flow in the

stress ulcer. XXII Autumn Meeting of Jpn Soc of Gastroenterol, Oct 1980

Additional comment 3) Changes in blood

vessel and blood flow with regerd to

their invo lvement in the development of stress ulcer

Masatoshi WATANABE 3rd Dept Intern Med, Jikei Univ

Sch Med, Tokyo

The investigation was made on the involvement of autonomic nervous system in blood flow change in the fine gastric mucosal vasculation.

Method: Wistar male rats used. Experimental ulcer was produced by giving stress through forced restraint and immersion in water. As autonomic drugs were used representative adrenergic drugs-- epinephrine (Epi), norepinephrine (NE), isopro- terenol (Iso), adrenergic blocking agents--phenoxy- benzamine (POB), propranolol (Pro), prazosin (Praz).

Results: 1) When Epi, NE and Pro were given

after stress, no changs was observed in the incidence of ulcer. After the administration of Iso, POB and Praz, the incidence decreased. 2) As for change in blood vessel seen in the transparent preparation, Epi

and NE induced narrowing of the lower part of the central vein on the muscularis mucosa, and ischemia and congestion in deeper and superficial layers of mucosa respectively. After Iso, POB and Praz, however, none of these changes was observed. 3) Ad-

renergic drugs reduced gastric secretion under stress. 4) Epi and NE decreased blood flow in gastric mucosa.

Discussion: It was considered that anti-ulcer effect of autonomic drugs might attributed to the inhibition of blood flow disturbance.

References: 1) Sugiura M, et al: Studies of microcirculation by transparent preparate using cooled

methanol-fixation. JJpn Coil Angiol 9: 428, 1969.2) Mural S, et al: Arterio-venous anastomoses in gastric mucosa and their significance. Jpn J Gastroen- terol 76: 2056, 1976. 3) Watanabe M, et al: Rela- tion between autonomic nervous and changes of gas-

4. Study on in t ra lumina l blood flow as the etiological factor of

ischemic colitis

Koji NOMURA and Yoshihito UCHIDA 1st Dept Intern Med, Yamaguchi Univ

Sch Med, Ube

The purpose of this study was to investigate an as- sociation between intraluminal pressure and colonic

blood flow, which might be involved strongly as the genesis of ischemic colitis.

As the control study, the blood flow in normal mucosa was measured in contracted and dilated sig-

mold colon using healthy volunteer, while it was also estimated in the mucosa bearing several grades of

ischemic colitis (active, healing and scarred stages). Furthermore, using a rabbit, we studied on the blood flow in the segmented large intestine by ligation after insertion of intraluminal catheter into the colon. The mucosal blood flow in both sites of taenia and non-taenia coli was measured in the same time when segmented intestine was inflated following stepwise increments of intraluminal pressures of 0, 10, 30 mmHg and following decompression.

As the results, there was no statistical difference of the blood flow between contracted and dilated colons in the control study. The blood flow in the cases of ischemic colitis decreased in active stage and reversely increased in healing stage as compared

with it in the mucosa apart from involved area. However, in scarred stage there was no difference

among the measured area in the entire colon. In the rabbit, the changes of mucosal blood flow measured in several conditions inflated with intraluminal pres-

sures of 0, 10, 30 mmHg and following decompression were as follows; (1) at the site of taenia coli:

97.5, 105.9, 10.3, 99.5 ml/min/100 g respectively, and (2) at the site of non-taenia coli: 83.4, 94.8, 32.9, 89.1 ml/min/100 g respectively. With the pressure of 30 mmHg, the blood flow in both sites was reduced, but especially the reduction at the site of taenia coli was markedly. After decompression to


0 mmHg, the blood flow re tu rned to almost the

same level in bo th sites.

Colonic mucosal blood flow was clearly infleenced

by the change of in t ra lumina l pressure, showing

that part icular ly the blood flow of the site of taenia

coli decreased much more by the h igh pressure. This

may be one of the reasons why longi tudinal ulcer is

seen frequently at the site of taenia coll.

In conclusion, it is strongly considered that the

in t ra lumina l pressure might be involved into the

etiological factor of ischemic colitis.

5. Cl in ica l and pathophysiological study of ischemic colitis

H. SHIGEMATSU and K. SASAKI

1st Dept Surg, Univ of Tokyo, Tokyo

Included in this study are 21 men and 4 women

with an average of 68 years. T he vast majori ty of pa-

tients had systemic arteriosclerotic diseases. The dis-

t r ibut ion of colonic lesions differs from a previous

reports in the higher incidence of involvement distal

to the splenic flexure (21 cases). T he splenic flexure

was involved in only 4 cases. A series of experiments

was performed in dogs whose inferior mesenteric

and the lowermost l u m b a r arteries were divided and

the terminal abdomina l aor ta was ligated. These

arterial l igation caused the submucosal blood flow

in the distal colon to decrease by 59%, and resulted

in t ransient ischemic colitis. T he release of a c lamp

on the graft bypassed the site of l igation of the ab-

dominal aorta caused a decrease in submucosal

blood flow in the distal colon by 45 % in contrast to a

decrease inflow th rough the superior mesenteric

artery by only 9%. These hemodynamic alterations

in the left side colon may play an impor t an t role in

the ischemic colitis.

Al though the severity of the insult of the colonic

wall may be determined by in t r amura l blood flow to

the colon, it is very difficult to establish the presence

of poor blood flow to the involved colonic wall in a

h u m a n patients with ischemic colitis. To date the

term of "ischemic colitis" is eventually a clinical

diagnosis based on the clinical presenta t ion and sub-

sequent course.

Addit ional comment 4) Parameters for the

assessment of recovery pattern i n

ischemic bowel , wi th special reference to the mucosal blood

flow in the intestine

M. O G I W A R A and S. HAYASHI

Dept Surg L Shinshu Univ Sch Med, Matsumoto

The present study was designed to de termine

whether the recovery pa t te rn of the mucosal blood

flow, est imated by e ther the measurement of intra-

luminal tempera ture , heat clearance or hydrogen

gas clearance technique, might be valuable for pre-

dict ing heal ing process in the ischemic mucosa,

caused by the temporary occlusion of the proximal

por t ion of the superior mesenter ic artery.

In dogs with intestinal ischemia for 30 minutes in

dura t ion, indicat ing a rapid recovery in the heal ing

process on the intestinal mucosa, the mucosal blood

flow, est imated by any kind of methods for measure-

ment , was rapidly re tu rned toward the control

values within ten minutes af ter release of the occlu-

sion. On the contrary, in groups with ischemia for

one to four hours, associated with an evident

damage on the mucosa, mucosal blood flow was not

re turned toward the control level within one hour

after release of the occlusion. Al though the hydro-

gen gas clearance technique might be evaluated for

indicat ing an absolute value of the blood flow, the

measurement of in t r a lumina l t empera ture and the

heat clearance technique might also be useful to il-

lustrate the recovery pa t t e rn as to the mucosal blood

flow, predict ing the consequent heal ing process in

experiments with intest inal ischemia.

Reference: 1) Ogiwara M: Shinshu Med J 29:

687-706, 1981

Addit ional comment 5) Microangiographic studies in non-specific ulcerative

lesions of the intestine and

Neoplastic tumors of the colorectum

Takatoshi SHIMOYAMA and Masao T O M I T A

Dept Surg, Nagasaki Univ Sch Med, Nagasaki

156 Proceedings of24th Autumn Meeting Vol. 18, No. 2

Morphological analysis of non-specific ulcerative

lesions of the intestine and neoplastic tumors of the

colorectum was examined by microangiography re-

lating to their pathogenesis. The material consisted

of intestinal specimens taken at operation from 3

patients with ulcerative colitis, 4 patients with

Crohn's disease, 28 patients with neoplastic polyps

and 108 patients with 115 advanced cancers of the

colorectum. 1) In ulcerative colitis there were hyper-

vascularity in the mucosa and submucosa, but the

muscularis propria and serosa maintained normal

vascular patterns even though they were dilated and

slightly scrambled. However, these patterns varied

with degree of severity of the morphological

changes. Vascular patterns of pseudopolyposis were

classified into 2 types; a 'reef-like' appearance sup-

plied from large penetrat ing vessels and numerous

straight vessels of the submucosa, and mucosal vas-

cular patterns in the remaining mucosa. Vascular

congestion with edema and vasculitis of the capillary

vessels were seen in the remaining mucosa. 2) Vas-

cular patterns of Crohn's disease demonstrated more

diffuse hypervascularity in all layers in connection to

lymphoid follicles and deep transmural lymphatic

infiltrate. In areas with aphthoid ulcer, there was

marked focal hypervascularity in the mucosa with

numerous dilated small vessels. In a case with severe

vasculitis of the colonic wall, scrambled vessels of all

layers and less decreased penetrating vessels were ob-

served. These vascular changes might suggest the

possibility of pathogenesis of these lesions. 3) The

vascular patterns of adenomatous polyps were clas-

sified into 4 groups and had been discussed with de-

grees of dysplasia. The advanced cancers showed

vascular disarray with vessels of varied size which

histological foci of necrosis were often seen. A good

correlation between the degree of vascular involve-

ment and their plasma CEA level was observed.

References: 1) Morson BC, Dawson IMP: Gastro-

intestinal Pathology, Bluckwell Scientific Publica-

tion, Oxford, 1972. 2) Shimoyama T, et al: J Jpn

Surg Soc 82: 169, 1981

6. Variations of mucosal and submucosal blood flow and activity of autonomic

nerves in the formation and healing of gastric ulcer

Tetsuo M O R I S H I T A and Masahiko NAKAMURA

Dept Intern Mecl, Sch Med, Keio Univ, Tokyo

This study was performed to investigate the influ-

ence of the gastric blood flow and sympathetic or

parasympathetic nerves on the formation, aggrava-

tion and healing of gastric ulcer.

The gastric mucosal blood flow, estimated with

hydrogen clearance method, around the restraint-

induced ulcer (42 +_ 15 ml /min /100 g, n=9,

0.005<p(0.01) and acetic acid-induced ulcer (34 +_

15, n=9, p<0.005) were significantly decreased as

compared to controls (89 + 37, n=24). In 8 patients

with acute gastric ulcer, 6 patients (75%) showed

the maximal submucosal blood flow at the endo-

scopic active stage, A1 or A2, and 7 patients (88%)

showed a peak of the increase at the healing stage,

HI , with the gradual decrease toward the complete

healing. Among 12 patients with intractable ulcer, a

peak O f the increase was seen at the acute stage, A2,

in 6 patients (86%) and at the healing stage, H2, in

7 patients (100%) of 7 patients who were successfully

treated. They were 2.4 + 0.67 and 2.0 + 0.63 fold

increase respectively compared to the submucosal

blood flow at each initial examination. There was a

decrease of 54 ___ 22% of the submucosal blood flow

in 7 (78%) of 9 episodes of the enlargement of ulcers

in 5 patients who were unsuccessfully treated. AChE

activity (modified Karnovsky-Root's method) in-

creased with the decrease of sympathetic nerve ac-

tivity (Farck-Hillarp's method) at the margin of the

rat restraint-induced ulcers and human gastric

ulcers.

It is concluded that: 1. The formation and aggra-

vation of gastric ulcer are associated with 50% de-

crease of mucosal and submucosal blood flow and

stimulated parasympathetic nerve. 2. In the healing

course of human gastric ulcer, a peak of the increase

of the gastric submucosal blood flow was seen at the

endoscopic H1 stage in acute gastric ulcer, and A2

and H2 stages in intractable ulcer without the

gradual increase.


Additional comment 6) Measurement of

regional gastric and duodenal mucosal blood flow in chronic gastric, duodenal and polypectomy-

induced ulcer patients

Motonobu MURAKAMI* and Takeo MIYAKE**

*Dept Geriat Med and

* *lst Div Intern Med,

Fac Med, Kyoto Univ, Kyoto

Regional mucosal blood flow in chronic gastric

ulcer, duodenal ulcer and polypectomy-induced

ulcer patients was studied: Normal subjects who

were confirmed to be free of ulcer by endoscopic ex-

amination were used as a control. Mucosal blood

flow was measured using the contact electrode and

hydrogen clearance method. In the control group,

regional gastric mucosal blood flow in the proximal

side of the angle was lower than other sites of the

stomach. Mucosal blood flow of chronic gastric

ulcer of the angle and corpus showed a decrease in

the active stage of the disease and an increase with

progression of healing. In contrast, in polypectomy-

induced ulcers, there were no decrease in mucosal

blood flow during their active stage compared to

both the control group and surrounding mucosa.

Regional blood flow of the anterior wall of the duo-

denal bulb was lower than other sites in the control

group. In duodenal ulcer patients, regional blood

flow of the margin of ulcer and scarred area was

lower than that of the corresponding site of the con-

trol group. These findings suggest that regional

mucosal blood flow is involved in the mucosal de-

fence mechanism against peptic ulcer in the

stomach and anterior wall of the duodenal bulb.

Additional comment 7) Gastric mucosal hemodynamics and peptic ulcer

Nobuhiro SATO and Takenobu KAMADA

1st Dept Med, Osaka Univ Med Sch, Osaka

The mucosal blood flow in the stomach is an im-

portant factor in the etiology and healing of the gas-

tric ulcer. However, there have been no data avail-

able about the gastric mucosal hemodynamics in the

active ulcer and in its healing process in human.

Gastric mucosal hemodynamics and oxygen suffi-

ciency were studied in 42 patients with gastric ulcer

at angular portion of the stomach by reflectance

spectrophotometry performed during gastrofiber-

scopy. Spectra were taken at 20 different portions

throughout stomach, Fifty-one patients with no gas-

tric lesion were chosen as controls. The oxygen suffi-

ciency of gastric mucosa was assessed by determining

spectrophotometrically the oxygen saturation of

hemoglobin in the mucosal capillary blood. In pa-

tients with active gastric ulcer, the mucosal blood

volume decreased significantly at most regions in the

stomach. With healing of the ulcer, the mucosal

blood volume increased to the level of the control.

At the margin of the ulcer, the mucosal blood

volume increased by 40 50% as compared to that of

other portions during the healing stage and returned

to the control level at the scarring stage. The oxygen

saturation of Hb was not changed in the presence of

gastric ulcer. It is concluded that the gastric muco-

sal blood flow changes during the healing processes

of the ulcer, which would influence the oxidative

metabolism of the mucosa and healing of the ulcer.

7. Portal hemodynamics and gastric mucosal blood flow of hemorrhagic gastritis

in por ta l hypertension

Atsushi T O Y O N A G A

2nd Dept Intern Mecl, Kurume Univ

Sch Med, Kurume

One hundred eighty-six cases of portal hyperten-

sion, 155 liver cirrhosis, 19 liver cirrhosis + hepato-

cellular carcinoma, 10 idiopathic portal hyperten-

sion and 2 primary biliary cirrhosis, were selected

for the study. Splenoportographic, angiographic

and percutaneous transhepatic photographic ex-

aminations of portal hemodynamics were under-

gone in men, while gastric mucosal blood flow was

examined by postmortem microangiography, his-

tology or IlSLMAA and radioactive microsphere

(Sr ss labelled 15/t and Ce 141 labelled 50// tracer

microsphere) in portal hypertensive dogs produced

by entire liver compression technic. The back-

ground of hemorrhagic gastritis in portal hyperten-

sion was (1) hyperhemodynamic state of upper gas-


tric mucosa, (2) marked development of gastric

arteriovenous shunting and (3) markedly increased

and congested blood flow in the upper gastric

mucosa. Consequently hemorrhagic gastritis in por-

tal hypertension would be frequent in incidence and

severe in clinical course.

References: 1) Inokuchi K: Study on portal

hypertension. JJSS No 79, Vol 12, 1978.2) Rudolph

AM, et al: The circulation of fetus in utero. Circula-

tion Research. Vol 21, Aug 1967. 3) Yamana H, et

al: Hemodynamic studies on the development of

esophageal varices in the portal hypertensive dog.

JJSS No 81, Vol 12, 1980. 4) Fukuda K, et al: Cor-

relation between endotoxemia and endoscopic find-

ings of esophageal varices. Jpn J Gastroent Vol 76,

No 9, 1979

Additional comment 8) Experimental study on effect of estrogen to gastric

submucosal blood flow in

l iver cirrhosis

Akitake HASUMI and Haruo AOKI

Dept Surg, Sch Med, Fujitagakuen Nagoya Hoken Eisei Univ, Aichi

To investigate the effect of estrogen to micro-

hemodynamics of the stomach in liver cirrhosis, sub-

mucosal blood flow, submucosal oxygen tension and

number of A-V shunts of gastric wall have been

studied in three groups of male rats: rats in A group

are administered estradiol intramuscularly (total

doses 80 m g / 8 weeks), rats in B group are made liver

cirrhosis by Asahikawa Univ. method, rats in C

group are made liver cirrhosis and administered

estradiol.

Both submucosal blood flow and oxygen tension

at the corpus of the stomach increased remarkably

in C group comparing with those of A and B group.

Number of A-V shunts also revealed significant in-

crease in C group,

It is suggested that the increase of blood flow, oxy-

gen tension and number of A-V shunts in sub-

mucosa at the upper part of stomach in rat with liver

cirrhosis are caused by hyperestrogenemia and that

the regional hypertension is observed. This is our

another concept of pathogenesis for the esophageal

varices in portal hypertension in addition to the

traditional concept of hepatofugal collateral.

Additional comment 9) Role of blood flow in the development of gastric mucosal injury associated with hepatohiliary

and pancreatic diseases

Tadao MANABE and Takayoshi T O B E

1st Dept Surg, Fac Med, Kyoto Univ, Kyoto

The mechanism of gastric mucosal injury asso-

ciated with necrosis of the liver, cirrhosis of the liver,

obstructive jaundice and acute and chronic pan-

creatic diseases was studied from a hemodynamic

point of view using a radioactive microsphere tech-

nique in rabbits.

In rabbits with the necrosis of the liver, obstruc-

tive jaundice and acute pancreatitis, a significant

decrease of blood flow in every part of the stomach

was observed in the mucosal layer and numerous

erosions appeared in the corpus. The higher sus-

ceptibility of the gastric mucosa in these diseases

may be due to lowered mucosal blood flow in the

stomach.

In rabbits with cirrhosis of the liver and chronic

pancreatitis, a marked increase of the mucosal

blood flow was observed in the mucosal layer of the

corpus and erosions or shallow ulcers appeared in

the antrum. Since the mucosal blood flow increased

in the corpus in tetragastrin-induced hypergastrine-

mia, the hemodynamic pattern of the stomach with

cirrhosis of the liver and chronic pancreatitis was

suggested to be caused by the greater degree of

secretagogue.

Thus, the changes of the mucosal blood flow was

considered to play an important role in the develop-

ment of mucosal injury in the hepatobiliary and

pancreatic diseases.

8. Measurement of hepatic tissue blood flow (HTBF) by the hydrogen gas

clearance method under laparoscopy

Takamitsu MAKINO and Hiroshi ISHIDA

3rd Dept Intern Med, Nippon Med Sch, Tokyo


A method to measure regional hepatic tissue

blood flow (HTBF) by hydrogen gas clearance method was described. HTBF was determined at a phase when hydrogen gas contribution from portal

vein became negligible and the clearance curve became monoexponential. In rabbits whose hepatic artery was ligated, there existed a good correlation

between HTBF obtained by hydrogen gas clearance method and portal blood flow measured by electro- magnetic flowmeter.

HTBF in 4 controls was 82.3 + 25.78 ml/min/100 g (M _+ SD). Data obtained in various liver diseases were followings a) acute hepatitis-acute stage (n=4) 40.15 -+ 13.22, b) -recovery stage (n=3) 87.23 _+ 24.79, c) chronic inactive hepatitis (n=22) 51.44 _+ 15.06, d) chronic active hepatitis (n=16) 41.64 +

12.93, e) In liver cirrhosis (n=33), HTBF varied de- pend on the site of electrode insertion, it was 45.31 + 13.12, f) HTBF in fatty liver (n=5) was 27.54 +

2.87. The apparent low value in this disorder was

considered to be resulted from low setting of blood/tissue partition coefficient, g) In scar liver (n=4), there existed significant difference in HTBF

between residual parenchyma and scar (60.33 _+ 19.65 vs 27.20 _+ 3.18), h) In hepatoma (n=5), HTBF in tumor was 77.25 _+ 2.80 and that in intact area was 50.13 _ 4.57, i) In metastatic liver carcinoma (n=7), HTBF in tumor was 31.47 + 3.48 and that in intact area was 57.77 + 5.55.

The method described is unique to provide data

on regional tissue blood flow and thus could offer useful clinical informations in assessing various liver disease.

9. Gastro-intestinal blood flow and free- radical in hyper-coagulable state

Toshikazu YOSHIKAWA and Keiji FUKUMOTO

1st Dept Med, Kyoto Prefect Univ Med, Kyoto

Hemorrhagic lesions in the small intestinal mucosa have been demonstrated in humans and experimental animals following hemorrhagic shock, intestinal ischemia and disseminated intravascular coagulation (DIC). In order to define the role of free-radicals in the pathogenesis of the mucosal

lesions, we examined thiobarbituric acid (TBA) reactive substances resulted from lipid peroxidation, lysosomal enzyme beta glucuronidase, and many coagulation tests in experimental model of DIC, hemorrhagic shock and intestinal ischemia.

TBA reacting substances in the small intestinal mucosa were increased significantly in rats or dogs experimental DIC, hemorrhagic shock, and intes-

tinal ischemia induced by the ligation of mesentric

artery. And also, beta glucuronidase activities were

increased after the onset of these ischemic conditions. From these data, we can suggest that free- radical lipid peroxidation are responsible for the intestinal mucosal damage produced by ischemia.

Additional comment 10) Gut microcirculation and coagulation-

fibrinolysis system

Akimasa NAKAO and Tatsuhei KONDO 2nd Dept Surg, Nagoya Univ Sch Med, Nagoya

Pathologic derangement of microcirculation in the gut and its relation with coagulation and fibrinolysis system in the tissue of the gut and blood were investigated experimentally in mongrel dogs.

Acute portal vein occlusion (PVO) induced intrin- sic and extrinsic hypercoagulation in portal blood

because of congestion in the portal bed. Within 10 minutes after PVO, fibrin deposited in the mucosal small vessels of the gut, which process suggested that

disseminated intravascular coagulation (DIC) occurred. Tissue plasminogen activator activity, which plays important role in lysis of fibrin clot, located mostly in the endothelial cells of small vessels in submucosal layer, and disappeared within 20 minutes after PVO, resulted in irreversible DIC. ELT in the portal blood was shortened remarkably after PVO, probably due to release of tissue plasminogen activator into portal blood, which was shown by depres- sion of tissue plasminogen activator activity.

When PVO and ligation of the superior mesenteric artery were performed simultaneously, these changes were postponed.

To the contrary, when portal bed was decom- pressed by bypass between portal vein and femoral vein with the heparinized hydrophilic catheter, these

160 Proceedings of 24th Autumn Meeting Vol, 18, No. 2

derangement of coagulation and fibrinolysis system

was not observed.

Thus, tight relationship between microcirculation

of the gut and coagulation and fibrinolysis system

were clearly demonstrated.


Gastric Cancer and I m m u n o l o g y wi th Special R e f e r e n c e to I ts

Modal i ty and T r e a t m e n t

C o - M o d e r a t o r s : Dr . T a k a o H A T T O R I and Dr . Ak i r a Y A C H I

1. Cl inical significances of tumor antigen- specific suppressor T cells from gastric cancer patients, and phenotypic cell

surface analysis on a fluorescence- activated cell sorter (FACS-1V)

using monoclonal antibodies

Shohei KOYAMA* and Katashi FUKAO* *

�9 Depts Intern Med and **Surg,

Inst Clin Med, Univ of Tsukuba, Ibaragi

We demonstrated the presence of suppressor cells

from peripheral blood lymphocytes and spleen cells

in gastric cancer patients by the functional assay

established in our laboratory. The immunosuppres-

sor cells in our system are (1) "T" suppressor cells,

(2) tumor antigen-specific, and (3) associated with

the presence of tumor. The suppressor T cells acti-

vated by tumor cells in cancer patients were success-

fully grown by the use of T cell growth factor

(TCGF) prepared from human tonsils or spleens.

Immunological phenotypic cell surface analysis was

performed by direct or indirect immunofluorescence

staining using a monoclonal (anti Leu-1, anti Leu-

2a, anti Leu-3a, anti Leu-4, anti Leu-5, anti Leu-7,

anti Human H L A - D R and human anti-Ig) anti-

bodies. Quantitative fluorescence measurements

were made on FACS-1V (Becton Dickinson, FACS

systems, Mountain View, Calif.). Functional sup-

pressor T cells expanded with TCGF showed a

phenotype Leu-1 +, 2a +, 3a - , 4+, 5 +, 7 - , HLA-

DR + and I g - .

These findings suggest that suppressor T cells ac-

tivated by tumor antigen observed in our study may

exert negative control in the immune response, thus

inducing the status of the lower cell-mediated anti-

tumor immunity, and may promote cancer progres-

sion in gastric cancer patients,

2. I m m u n e response to the growth of human cancer in the gastro-

intestinal tract

Akira AOIKE and Yoshinori T A N A K A

Dept Prevent Med, Kyoto Prefect Univ Mecl, Kyoto

Natural cytotoxicity against Namalva target cells

was determined in the peripheral blood of healthy

donors and gastric cancer patients by specific 51Cr

release assay. The NK cell activities of blood lym-

phocytes from normal healthy donors (1 1.5 + 7.6%)

were higher than those from gastric cancer patients

with stage 1 (5.1 _+ 3.6%) and stage 4 (3.4 ___ 1.6%)

but were similar to those from patients with stage 2

(9.8 _+ 6.9%) and stage 3 (13.5 ___ 9.0%). We then

measured interferon titers in sera of healthy donors

and patients with stage 1 and stage 4 was almost

negative, but in patients with stage 2 and 3 it was

346 _+ 281 and 253 ___ 202 i .u . /ml respectively. We

also proved that co-culture of tumor cells with peri-

pheral blood lymphocytes could produce IF in the

medium in vitro and so it is easy to suppose that the

same phenomena may be taken place in vivo. The

findings of higher NK activities in gastric cancer pa-

tients with stage 2 and stage 3 than those with stage

1 and stage 4 may be attributable to high IF titers in

patients with stage 2 or stage 3. Then the percentage

of peripheral blood lymphocytes reactive with

monoclonal antibodies identifying T cells (T3+),

helper / inducer cells (T4 + ) and suppressor/cytotoxic

ceils (T8 +) was determined in young, aged humans

and gastric cancer patients, The T4 + and T8 + cells

in the aged persons were significantly lower than the


young men. The T3 +, T4 +, and T8 + cells showed

no differences among early and advanced gastric

cancer patients.

3. N K activity in patients wi th cancer of digestive organs and

effects of IL-2

Yoshihiro ABE* and Fujiro SENDO**

*2nd Dept Intern Med and

**lst Dept Pathol,

Yamagata Univ Sch Med, Yamagata

At first, we tested NK activity of cancer patients

with digestive organs by using fresh peripheral blood

lymphocytes as effector cells. When we compared to

NK activity of all tested patients with that of control

healthy subjects, we could find statistically only a

slight tendency of decrease of the NK activity in the

patients with cancer. However, if we divide the can-

cer patients into two groups, either operable or in-

operable, the NK activity of inoperable patients is

significantly lower than that of healthy controls. The

NK activity in the majority of the patients with can-

cer of digestive organs was augmented by addition of

a kind of lymphokine, IL-2 into the culture media.

However, the rate of augmentat ion was lower than

that in the control after the same treatment. Al-

though the NK activity of fresh peripheral blood in

the patients with operable cancer was within the

range of healthy subjects, the NK activity after the

treatment by IL-2 in the above patients was also

lower than that of healthy control.

Addit ional comment 1) Cytotoxic activity of patients with advanced

gastric cancer

Morihiro OHARA and Reiji KASUKAWA

2nd Dept Intern Med, Fukushima Med Coll, Fukushima

Natural cytotoxicity (NC) of peripheral lympho-

cytes, monocytes and polymorphonuclear leukocytes

(PMN) of patients with advanced gastric cancer

measured by means of 51Cr releasing assay against

K562, KATOII I and PC1.

The NC activity of PMN in the patients against

K ATOII I and PC1 was higher than that of normal

PMN.

The NC activity of monocytes and PMN in the pa-

tients against KATOII I and PC1 was stronger than

NK activity of the lymphocytes.

NK and ADCC activity of lymphocytes infiltrated

into gastric cancer tissue measured by means of simi-

lar assay against K562 (NK) and P815 (ADCC).

The NK and ADCC activity of lymphocytes infil-

trated into surrounding area of cancer tissue was

stronger than that of lymphocytes infiltrated into the

cancer tissue and the normal area of the tissue.

4. The nonspecific immunosuppression in cancer patients of GI tract

Yutaka KOHGO and Ichiro URUSHIZAKI

4th Dept Intern Med, Sapporo Med Coil, Sapporo

The nonspecific immunosuppression in patients

with gastrointestinal malignancies were examined

from the points of humoral and cellular inhibition.

(1) Humoral inhibition: Sera from cancer patients

inhibited the response of blastogenesis or lympho-

cytes by PHA, 2-way mixed lymphocyte reaction

and natural killer cell activity. The characteristic

human serum component which inhibited the im-

mune response in vitro was acidic alpha-2 macro-

globulin fraction by successive chromatographic

procedures. The enzyme linked immunoassay for

this acidic alpha-2 macroglobulin fraction showed

higher values in malignancies and its values were

pararelled with diseased stage. (2) Nonspecific cellu-

lar suppression: Adherent cells from cancer patients

inhibited 2-way mixed lymphocyte reaction and the

suppression was mediated by secretion of prosta-

glandin from suppressor macrophage. The suppres-

sor macrophage also impaired the production of

Interleukin-2 in cancer patients. (3) Elimination of

serum suppressive factors by plasmapheresis: Plas-

mapheresis of patients with advanced gastric car-

cinoma was effective for improvement of host im-

mune response such as PPD skin test, and decrease

of serum inhibitory factors.


5. Analysis of an t i - tumor i m m u n e reac-

t ions in patients with hepatoma

Yasutaka INAGAKI and Masaharu TSUCHIYA Dept Intern IVied, Sch Med, Keio Univ, Tokyo

We investigated humoral and cellular immune

reactions to hepatoma cells in 32 patients with hepatoma. NK activity of peripheral blood lymphocytes (PBL) against K562 was significantly decreased in the patients compared with healthy controls.

Specific cytotoxicity of PBL against HCC-M (human hepatoma cell line) was not observed in a autologous patient as well as allogeneic patients who shared at

least one of HLA A B C antigens of HCC-M. How- ever, specific immune reaction to hepatoma was de-

tected by repeated micro-LAI assay. Anti- tumor

antibody was detected by 12sI labeled protein A assay and ADCC assay.

Then we tried to generate the cytotoxic lymphocytes against HCC-M using PHA and Interleukin 2 (IL-2). Cytotoxic activity of PBL stimulated with PHA was enhanced against HCC-M and CCRF- CEM but not against normal PBL. In one out of 7 patients, PBL cultured with IL-2 showed markedly enhanced cytotoxicity against HCC-M but not against HeLa and S-170 (human skin fibroblast). Ninety eight percent of the lymphocytes cultured with IL-2 showed conventional T cell surface marker (E rosette formation). These cultured lymphocytes may be effective in the adoptive irnmuno-

therapy of human hepatoma.

7. Immunolog ica l studies on the gastric

cancer - - w i t h special reference to

stage, course, sp lenectomy and

i m m u n o m o d u l a t o r - -

K. YOSHINO, K. KUMAI and

F. ASANUMA Dept Surg, Keio Univ Sch Med, Tokyo

This communicat ion describes briefly the results obtained from the immunological studies on gastric cancer in order to evaluate the role of immune in the patients with gastric cancer. For this purpose following evaluation was performed. Various nonspecific immunological parameters, i.e. PPD, PHA skin

test, number of lymphocyte and T / B cell subpopu~ lation in peripheral blood, IgG Fc receptor positive T cell, fraction of serum protein, immunoglobul in

and complements in serum, were compared according to stage of the cancer and pre and post-operative

course. The rationale of splenectomy, which has

been performed chiefly to remove the metastasized lymph nodes in splenic hilus, was re-evaluated by

clarifying the incidence of the metastases in the nodes, by postoperative survival rate and by the immunological parameters. The effectiveness of the immunomodulators, i.e. OK-432, Levamisole, Len- t inan and Bestatin was evaluated by means of PPD skin test and durat ion of postoperative disease free

interval. The following conclusions appear justified: (1)

with aggravation of the stage of the cancer, immunological potency seemed to fall down, (2) after curative resection, immunological potency seemed to fall down at the 1st week postoperatively and to require 3 weeks for recovery, (3) the incidence of

metastases in the lymph nodes in splenic hilus, which is able to be the incidence of justifiable

splenectomy, was revealed to be only 17.4 per cent, the survival of splenectomized group was shown to

be possible to be lower than that of nonsplenecto- mized group, and this tendency was recognized more markedly in the group of earlier stage, (4) the

number of platelets was shown to be higher in the splenectomized group postoperatively, and (5) with administration of the immunomodulator , PPD skin test was revealed to be raised up and the disease free interval elongated.

8. I n t r ade rm a l injections of the streptococcal preparat ion OK-432 for inoper-

able gastric cancer patients as effective i m m u n o t h e r a p y

Hitoshi HANAUE* and Satoshi YOSHIZAKI** *lst Dept Surg, Teikyo Univ Sch Med, Tokyo

* *Dept Surg, Fujita-Gakuen Univ Sch Med, ~qichi

Nonspecific immunotherapy with OK-432, peni-

cillin and heat treated lyophilized power of Su-strain of streptococcus pyogens A3, was evaluated in pa-

April 1983 Proceedings of 2 4th Autumn Meeting 163

tients with recurrent or unresectable stomach cancer

to assess the relative benefit of the preparation ad-

ministered by different routes. Comparative studies

were made of the variation in immunological para-

meters, the survival rate and the incidence of ad-

verse reactions in two groups of patients with uni-

form background factors: 24 receiving the prepara-

tion intradermally and 17 receiving intramuscular

doses of the preparation. In the former group, no

serious adverse reaction was found and more

marked improvement was achieved in different im-

munological parameters examined, particularly the

peripheral blood lymphocyte count, T-cell rate in

peripheral blood lymphocytes, a delayed hypersen-

sitivity skin reaction to polysaccharide extracted

from the cell wall fraction of the Su strain bacteria

of the streptococcus pyogens, and a agglutinin titer

against streptococcal cells. The survival rate was sig-

nificantly higher for patients receiving intradermal

than those receiving intramuscular doses of the

preparation.

Appointed comment 1) Immuno log ica l

competence in patients wi th gastro-

intest inal cancer and effect of

treatment on the immunolog ica l

parameters

Hisashi FURUE

Dept Intern, Teikyo Univ, Tokyo

Patients: Benign diseases 323 cases, malignant

diseases 266 cases including gastric cancer 78 cases,

colon cancer 15 cases, liver cancer 15 cases, bile duct

cancer 11 cases, and pancreas cancer 8 cases, and

healthy control 118 cases.

Immunological observations: Number and per-

cent of lymphocyte, active T cell, T cell, B cell, and

IgG Fc receptor positive T cell, lymphocyte blasto-

genesis stimulated by PHA or concanavalin A, IAP

(immunosuppressive acidic protein), IgG, A, M,

PPD skin reactions, CEA, alpha-fetoprotein, fer-

ritin, ADCC, virus interference factor(s) induced by

poly I:C, or PHA of leukocyte in vitro.

Results: From our clinical observation, immune

status is significantly impaired in patients with ad-

vanced cancer. Such impairment is also obvious in

patients with gastrointestinal cancer. The extent of

immunosuppression correlates with the stage of can-

cer and level of tumor burden. Our analysis also

shows close correlation between immune com-

petence and life span of the patients with advanced

gastric cancer. These immunosuppression can only

be reversed by decrease of the tumor burden, i.e.

surgical removal of the tumor, regression of the

tumor after cancer chemotherapy. Cancer chemo-

therapy impairs immunological status both in re-

sponder cases, and in non-responder cases similarly,

but these side reactions are only transient. After ces-

sation of chemotherapy, in responder cases, definite

immunological recovery is observed. On the con-

trary, immunotherapy can show only limited favor-

able influence on these immunological parameters.

Therefore, potent cancer chemotherapy is the most

significant maneuvers in the treatment of patients

with advanced cancer.


R e c e n t Views o n P r i m a r y B i l i a r y C i r r h o s i s

C o - M o d e r a t o r s : Dr . N o b u h i r o S H I M A D A a n d Dr. Rei j i K A S U K A W A

1. P r imary bi l iary cirrhosis in J a p a n u R e s u l t

of na t ional survey

Kyoichi INOUE and Hiroshi SASAKI

3rd Div Intern Mecl, Toyama Mecl and Pharmac Univ, Toyama

The first national survey of primary biliary cirrho-

sis (PBC) in Japan has been carried out by the sub-

committee on autoimmune hepatitis. The present

study is the result of analysis of 177 cases with PBC

which were collected from 75 institutes in Japan dur-

ing the period from 1975 to 1979. The patients were

selected for inclusion according to the following

diagnostic criteria. 1. Patients with histological lea-


tures of chronic non-suppurative destructive chol-

angitis, 2. Patients with serum positive for antimito-

chondrial antibody (AMA) and compatible with

PBC histologically, 3. Patients not investigated his-

tologically, but highly suspicious of PBC on the

grounds of AMA, clinical symptoms and course.

The 177 patients were divided into two groups,

according to absence or presence of symptoms.

Sixty-five patients were asymptomatic (a-PBC) and

112 were symptomatic (s-PBC). Patients in the a-

PBC group were diagnosed at a mean age of 50.4 _+

9.4 (M _+ SD) while in the s-PBC group symptoms

began at a mean age of 47.7 _+ 11.6. Ten of 65 pa-

tients with a-PBC and 8 of 112 patients with s-PBC

were male. In the sixty-five patients with s-PBC the

disease began with pruritus. Concerning the compli-

cating autoimmune diseases Sj6gren's syndrome was

most frequently seen in both groups. AMA was de-

tected in 57 of 63 patients (90.5%) with a-PBC and

96 of 104 patients with s-PBC (92.3%). Twelve pa-

tients with a-PBC became symptomatic within an

average of 26.2 _+ 15.7 month (6 to 57 months).

These 12 patients had presented jaundice, pruritus,

xanthoma or gastrointestinal bleeding. Thirty-two

patients with s-PBC died in an average of 51.6 +

29.2 months. Most of these patients died of hepatic

encephalopathy and gastrointestinal bleeding.

2. The role of i m m u n e responses to bile

antigen in the pathogenesis of PBC and appearance of heterogeneic

mitochondrial antibodies in PBC and its related

diseases

Yasuro YAMAMOTO and Saburo ONISHI

1st Dept Intern Med, Kochi Med Sch, Kochi

Specific humoral and cellular immune reactions

against bile ducts may have a key role in the patho-

genesis of PBC where the destruction of septal and

interlobular bile ducts is considered to be the essen-

tial pathological event. As previously reported, we

prepared a protein fraction from human bile which

originated from the bile duct epithelium and bile

canaliculi by using gel-chromatography, and also

specific antibodies to them.

In patients with PBC, we investigated humoral

and cellular immune responses to this bile protein,

and localization of it in the liver specimen by using

indirect immuno-fiuorescent and immune-peroxi-

dase staining.

With the following results: 1) Cellular immune re-

sponses to bile duct antigen were observed in 18 of

19 patients with PBC by leukocyte migration inhibi-

tion test. 2) Humoral antibody to bile duct antigen

was detected in the form of immune complexes in

the sera of 9 of 13 patients with PBC by specific im-

munofluorescent staining of bile antigens in the

complexes bound to Raji cells. 3) On the liver sec-

tions from the patients with PBC, bile duct antigen,

immunoglobulin M and G and complement C3 and

C4 were detected respectively in the cytoplasm of

macrophages around the bile ducts and granuloma.

These findings were not observed on the liver section

from controls. This may indicate that bile duct anti-

gen and antibody complexes associated with com-

plements are phagocytized by macrophages.

Next, heterogeneity of mitochondrial antibodies

were examined in the sera of typical PBC and CAH-

PBC mixed type. M2 antibody (PBC specific anti-

body) was detected in all sera of 12 typical PBC and

of 2 CAH-PBC mixed type. M4 antibody (antibody

to trypsin-insensitive M4 antigen) was detected in

the sera of 2 of 11 typical PBC and all of 2 CAH-

PBC mixed type. Therefore analyzing of hetero-

geneity of mitochondrial antibodies may be a useful

tool to classify and understand chronic cholestatic

liver diseases.

3. Alteration of T cell subsets and clinical features in primary

bi l ia ry cirrhosis

Kiyoshi ISHIHARA and Masayoshi YAMASHIKI

3rd Dept Intern Med, Fac Med, Niigata Univ,

Niigata

For the purpose of analyzing immunological ab-

normality associated with primary biliary cirrhosis

(PBC), T cell subsets and suppressor cell activities in

peripheral blood lymphocytes were investigated in

20 patients with PBC and compared them with 14

normal controls. For this analysis of T cell subsets


we used monoclonal antibodies to a common T cell

antigen (T3) and to the surface antigens on helper

(T4) and cytotoxic/suppressor (T8) T cell subsets.

In addition, to clarify the immunological mecha-

nism of bile duct destruction, the lymphocytes infil-

trating into portal tract were characterized, by im-

munofluorescent technique with monoclonal anti-

bodies, described above.

Patients with PBC had significantly higher T 4 / T 8

ratio than healthy subjects. However certain pa-

tients showed to be low or normal ratio, who are

asymptomatic, mostly. When those patients were di-

vided into 2 groups by T 4 / T 8 ratio that consisted of

group 1 (ratio <3) and group 2 (-->3), the former

seemed to have higher level of the abnormality of

liver function and humoral immunity than the

latter. But mitogenic response was better reserved in

group 2 rather than in group 1. In patients with PBC, ConA-induced suppressor

activity tended to be more decreased than healthy

subjects. However no correlation of suppressor ac-

tivity to T 4 / T 8 ratio was observed.

A certain number of lyrnphocytes infiltrating in

periductal region were stained with OKT8-antibody

by IF techniques. The location of this subset indi-

cated that they played an important role in the im-

munopathogenesis of PBC.

4. Experimental primary bi l ia ry cirrhosis

induced in rabbits fo l lowing immuniza -

t ion with mucous membrane antigen of the gallbladder

Takeyuki MONNA and Takayasu MATSUI

3rd Dept Intern Med, Osaka City Univ

Med Sch, Osaka

Biopsy performed repeatedly in 28 individual rab-

bits during long-term sensitization with bovine gall-

bladder mucous membrane antigen revealed that

pathological conditions progressed in 14 rabbits, a

part of them showing a scarring of degree III ac-

cording to Sheuer's classification.

Of 8 cases which were examined for the copper

content of liver tissue, 5 cases of 63% gave a greater

value than 6.0 pg/dl .

The bile acid level in the gallbladder was lowered

in all 7 cases examined and, in contrast, that in

serum was elevated in 27 % of 22 cases.

At 10 weeks, the infiltrating lymphocytes pene-

trated through the basal membrane of bile ducts,

and got into the epithelial layer to cause degenera-

tion and necrosis of the ductal epithelium.

The degenerative changes of mitochondria and

emperipolesis by lymphocytes into epithelial cells

were recognized electron-microscopically.

Additional comment 1) Immunogenetic study of PBC

Hirotoshi MIYAMORI and Yasuhiro KATO

1st Dept Intern Med, Sch Med, Kanazawa Univ, Kanazawa

HLA typing was performed in 22 patients with

primary biliary cirrhosis (PBC). In PBC patients,

HLA-DR2 showed a statistically higher frequency

compared to the controls (68% v.s. 30%, corrected

X2=7,660, corrected P<0.042, RR=5.00), and also

HLA-Dwl2 (DHO) showed a statistically higher fre-

quency compared to the controls (69% v.s. 21%,

corrected X2=l 3,366, corrected P<0.03, R R = I 1.4).

Forty relatives of 8 patients with PBC were ex-

amined for liver function test, immunoglobulines,

autoantibodies and HLA antigens. Two of the eight

families had PBC patients in addition to propositus.

The first family contained two sisters with PBC, and

in the second family father and his daughter had

PBC. Mitochondrial antibodies were present in 10 %

of the forty relatives. There were no relationships

between the development of PBC and HLA haplo-

type.

These findings support the concept that the dis-

ease susceptibility may be genetically determined

and the environmental factors in addition to the

genetic factors may participate in the development

of PBC.

5. Studies on pathognomic progression of primary biliary cirrhosis (PBC)

by peritoneoscopy and ERC

Chihiro SEKIYA and Yasuyuki YAZAKI

3rd Dept Intern Med, Asahikawa Med Coll,

Asahikawa


Nineteen cases of PBC, which were diagnosed by

histology, AMA, ERCP and other method, were

studied by peritoneoscopy (esp. dye contrast

method) and ERC. The liver surface of PBC was

seen to be smooth or gently undulated in the perito-

neoscopic examination. But using the dye contrast

method, most of the PBC cases revealed a formation

of rough bigger blocks of l - 3 c m in diameter on

their surfaces. The convex areas appeared almost

normal. On the other hand, the concave areas con-

sisted of many small depressions of 0 .5 -2 .0mm in

diameter. According to the histologic findings, these

might be portal tracts affected by PBC. Some cases

of PBC had diffuse minute depressions which looked

as if they broke down rough bigger block into small

pieces. We called the former RBB group and the

latter DMD group. We investigated histologically,

symptomatically and biochemically, and concluded

that RBB group represented the early stage of PBC

and the DMD group the advanced stage of PBC.

We then examined the intra-hepatic bile duct by

ERC. Periductal oozing or dotted line appearance

were observed in the 9 cases of PBC, in which we

managed to get good contrast of the 5th or further

peripheral branches of bile duct. These findings

were not found in the 32 controls. So we thought

that PBC affected not only the small, but also the

larger bile ducts. And the larger ones might be con-

cerned with a formation of rough bigger block and

progression of PBC.

Additional comment 2) The characteristic finding of the liver surface of patients with primary biliary cirrhosis under

peritoneoscope, with a special reference to its undulated

surface

Morikazu ONJI, Yoshimasa YAMASHITA

and Yasuyuki OHTA

3rd Dept Intern Med, Ehime Univ Sch Medl Ehime

The liver surfaces of 6 patients with primary bili-

ary cirrhosis (PBC) was observed under a peritoneo-

scope. A gentle undulat ion is the common charac-

teristic peritoneoscopic finding of PBC.

Materials studied were 3 patients with asymto-

matic PBC (a-PBC), 2 patients with symptomatic

PBC (s-PBC) and one patient of the transient case

between a-PBC and s-PBC. Chronic non-suppura-

tive destructive cholangitis (CNSDC) were seen in all

cases. The three-dimensional reconstruction was

made with the 100 serial sections of 10/2 in the thick-

ness obtained from the part of the undulated area.

The gentle undulat ion was observed in all surfaces

of livers. The undulat ion seen in a-PBC was fewer in

number and lower in height than those of s-PBC.

Chronological observation revealed that the lobular

segmentation in the undulated areas progressed.

The three-dimensional reconstruction study of the

convexed part of the undulated area revealed that

the findings of the liver cell regeneration were

scanty, the fibrosis and the infiltration of the inflam-

matory cells were mild, and CNSDC was not found.

On the other hand, the same study of the concaved

part of the undulated area revealed that the pro-

liferated collagen fibers and the infiltration of in-

flammatory cells were remarkable and CNSDC were

found, and the number of interlobular bile ducts in

one portal area was significantly (p<0.005) de-

creased. According to those findings, the gentle un-

dulation is considered to be passively produced by

the atrophic changes of the portal area, including

the injured interlobular bile ducts.

Additional comment 3) Laparoscopic evaluation and clinical course of

primary biliary cirrhosis

Akira YOSHIBA

Dept Diges Sys, Toranomon Hosp, Tokyo

Our subjects are 17 patients who had been ad-

mitted to our hospital and diagnosed as PBC his-

tologically, chemically and immunohematological-

ly. Laparoscopy was performed on a total of 22 oc-

casions in these patients, and findings thereof were

evaluated.

1) Laparoscopic findings of early PBC (Stage I

or II) were characterized by the dull edges of the

liver, with large nodes on the surface of the liver seen

as undulation. In addition, the surface was of multi-

color, and so-called "red patches (brown maculae)"


were particularly remarkable. The undulation and

brown patches were considered to be specific to early

PBC. 2) Out of 15 cases followed-up for 3 years or

longer after diagnosis, 4 ended in death: one died

after a year and 5 months, another died after 5 years

and 10 months and one more died after 6 years, all

from liver failure. The other patient died 6 years

after the diagnosis, from the complication of pan-

creatic cancer.

3) The case of asymptomatic PBC, which we

have followed for about 20 years and which pro-

gressed to symptomatic, may suggest the usefulness

of laparoscopy and may be very interesting in in-

vestigating this disease.

4) Our conclusion is that laparoscopy is useful in

the diagnosis of early PBC.

Additional comment 4) Significance of laparoscopic findings of the l iver

in the diagnosis of asymptomatic primary biliary cirrhosis

Yuzo MINAMI and Kouichi SEKI

2nd Dept Intern Med, Osaka Univ Med Sch, Osaka

Laparoscopic findings of livers of 14 patients with

primary biliary cirrhosis (PBC) including seven

asymptomatic and seven symptomatic cases were in-

vestigated. On the liver surface of asymptomatic

PBC, we found a new laparoscopic appearance,

consisting of reddish patches (1), which is irregular

in shape, ranging from 0.5 to 3 cm in diameter and

not evenly distributed. Six of seven cases of asympto-

matic PBC had reddish patches in our series.

From serial laparoscopic observations, the

patches were found to reduce the reddish tone and

to have protuberances with vaguely outlined white

markings in each area of them. The protuberances

and white markings are characteristic appearances

Df the liver surface in more advanced PBC cases.

The transition of reddish patches to protuberances

with white markings was in a coincidence with pro-

gression among histopathological stages in PBC

classified by Scheuer. In our histometrical examina-

tion, it can be considered that areas with reddish

patches had less advanced changes of intrahepatic

bile ducts and manifestation of reddish patches is at-

tributed to segmental progression and distribution

of histopathological changes in the liver with PBC.

As a result, we propose that "reddish patch" is a

new laparoscopic feature and has a significant value

in diagnosis for asymptomatic PBC.

Reference: 1) Minami Y, et al: Gastroenterologi-

cal Endoscopy 22: 503-514, 1980

6. Pathogenesis of primary biliary cirrhosis derived from ultra-

structural observations

Kazuo TOBE and Hideo NAGASHIMA

1st Dept Intern Med, Okayama Univ Med Sch, Okayama

The present study revealed bile duct lesions of

PBC ultrastructurally.

Materials and Methods: 52 interlobular bile ducts

from 10 patients with PBC were observed by TEM1).

39 bile ducts from 11 patients with other cholestatic

diseases were used as control.

Results: Many mononuclear cells infiltrated

among bile duct epithelial cells (BEC) of 9 ducts of

PBC. Some BEC were dropping out to the periduc-

tal area. Lymphocytes among BEC usually showed

activated features and extended their projections

onto BEC, some of which revealed degenerative

changes. At sites of emperipolesis, BEC showed

necrotic changes. These findings indicate lympho-

cyte cytotoxicity against BEC2). PBC epithelioid cells

were of the vesicle (sarcoid) type. BEC of 2 bile ducts

had numerous mitochondria (oncocytes). Periductal

capillaries had multilayered basement membranes

in all cholestatic diseases.

Conclusions: Cellular immunity was formed

against BEC in PBC. Mononuclear cell infiltration

and granuloma formation were probably caused by

dropped-out BEC antigens in the periductal area.

Damages of periductal capillaries might have con-

tributed to bile duct destruction in PBCS). Mito-

chondrial hyperplasia of BEC might have occurred

as compensation in the course of bile duct damage.

These mitochondria may trigger AMA.

References: 1) Tobe K: Acta Pathol Jpn 32: 57,


1982.2) Tobe K: Acta PatholJpn 32: 345, 1982. 3) Murakami T, et al: Arch HistolJpn 37: 245, 1974

7. Morphologlc studies on intrahepatic bile duct destruction in primary

biliary cirrhosis

Yasuni NAKANUMA and Goroku OHTA 2nd Dept Pathol, Sch Med, Kanazawa Univ,

Kanazawa

Bile duct lesions in PBC were examined by electron microscope and immunohistochemistry with emphasis on bile duct destruction. Electron micro- scopic studies in 16 patients with PBC disclosed four types of biliary epithelial injury undergoing destruc-

tion: 1) coagulative necrosis, 2) lyric necrosis without detachment from the biliary layer, 3) apoptosis

and 4) detachment of biliary epithelial cells from

the basement membrane and neighboring ceils. Lesions 1), 2) and 3) were also found in livers with

extrahepatic cholestasis. Lesion 4) was only observed in PBC, and might play some role in progressive bile

duct destruction in PBC. Migrating lymphocytes were often seen in the biliary layer, and some of them had pseudopod formation and were in close contact with injured biliary ceils. However, many severely damaged biliary cells were neither in contact with nor in the vicinity of migrating lymphocytes in PBC. Peroxidase method failed to demon- strate a deposition of both immunoglobulins and complements within periductal granulomas and at the basement membranes in 29 cases with PBC. Electron dense deposits suggestive of deposition of immune complex were absent within the basement membranes. Bile duct epithelial damages with periductal lymphoid cell infiltration and bile duct de-

struction respectively were found in 2 of the 5 cases with bone marrow transplantation and chronic GVHD, but their distribution was only focal.

From these findings, it seems likely that some of bile duct destruction are caused by cell mediated

cytotoxicity, but others seem unrelated to cellular immunity. Participation of deposition of immune complexes in the development of bile duct destruction seems unlikely.

8. The long term follow-up of primary biliary cirrhosis (P.B.C.): prog-

nostic factors, comparison of symptomatic and asympto-

matic cases

Yoshimi ITO and Masao OMATA 1st Dept Med, Chiba Univ Sch Med, Chiba

Clinical courses of P.B.C. were evaluated in 30 patients for mean follow-up periods of 45.5 months.

5 years survival of 20 patients with serum bilirubin

less than 2.0 mg/dl was 93%, whereas none of 10 patients with bilirubin greater than 2.0 mg/dl sur-

vived beyond 4 years. Similarly, 5 years survival of 19 patients with Scheuer 1-2 stage by the initial liver

biopsy was 92%, whereas none of patients with Scheuer 3-4 survived beyond 5 years. The presence

of increased orcein positive granules, marked peripheral hepatocyte ballooning and alcoholic hyaline

and absence of granuloma indicated poor prognosis. Thus, the serum bilirubin level greater than 2.0 mg/dl, liver biopsy of stage 1 and 2-Scheuer indi-

cated poor prognosis. After an average of 60 months of follow-up, all 12 asymptomatic patients were still alive and their serum bilirubin level stayed within 1.0 mg/dl. On the contrary, during an average of 40 months of follow-up, 11 of 18 symptomatic patients were dead and the mean serum bilirubin level of 18

symptomatic patients elevated to 6.0 mg/dl. 5 years survival of symptomatic P.B.C. was 32%. Impor- tantly, the survival rate of the asymptomatic patients did not differ from that of the general popula-

tion, matched by age and sex. The clinical course and prognosis of asymptomatic P.B.C. markedly differed from that of symptomatic P.B.C..

proceedings of the 24th autumn meeting from october 14–16, 1982-yamagata city, japan

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