CTHRC1 expression in primary biliary cholangitis.

OBJECTIVES: Collagen triple helix repeat containing-1 (CTHRC1) is a highly conserved extracellular matrix glycoprotein that is overexpressed in two murine models of cholestatic liver fibrosis. Elevated CTHRC1 has been found to attenuate liver fibrosis in these murine models, thus we aimed to study the expression of CTHRC1 in patients with cholestatic liver diseases and its correlation with hepatic conditions. METHODS: Ninety patients with chronic liver disease, including 48 had primary biliary cholangitis (PBC), 18 had primary sclerosing cholangitis (PSC) and 24 had chronic hepatitis B (CHB), together with five healthy controls (HC), were recruited to this study. Participants' liver sections were analyzed using immunohistochemistry. Serum CTHRC1 levels in another cohort of 59 patients with PBC and 10 age-matched HC were detected by enzyme-linked immunosorbent assay. RESULTS: CTHRC1 protein was primarily expressed in activated hepatic stellate cells (HSC). CTHRC1 expression was significantly increased in the PBC and PSC groups, compared with the HC and CHB groups. Importantly, the hepatic fibrosis stage of the PBC group was positively correlated with hepatic CTHRC1 expression (r = 0.425, P = 0.003). Meanwhile, there were significant correlations between serum CTHRC1 levels and both the degrees of hepatic inflammation and fibrosis stage in the PBC group (r = 0.300, P = 0.022; r = 0.321, P = 0.012). CONCLUSION: CTHRC1 may play a role in hepatic fibrogenesis in PBC and that serum CTHRC1 may be a potential novel noninvasive biomarker in the assessment of liver fibrosis and inflammation.

Microscopic features of small bowel mucosa of patients with Crohn's disease.

BACKGROUND: Double-balloon enteroscopy enables performing numerous small bowel biopsies for pathologic analysis. However, most histopathological characteristics of Crohn's disease are non-specific characteristics. We aimed to explore the small bowel mucosal histopathologic characters of Crohn's disease and identify some disease-specific changes. METHODS: We included 253 patients without tumors and grouped them into Crohn's disease, suspected Crohn's disease, and non-Crohn's disease groups. These patients underwent double-balloon endoscopy examination and small bowel biopsy at Renji Hospital, Shanghai. All histopathological sections were reviewed, and > 20 histopathological parameters were assessed. Immunohistochemistry was conducted when necessary. RESULTS: There were different forms of granulomatous lymphangitis on the small bowel mucosa in Crohn's disease. They showed as various macrophages or epithelioid cells in the lumina of lymphatics or in the center of the villi with or without evident obstruction. These features were only observed in Crohn's disease patients. Furthermore, they were correlated with granuloma and lymphangiectasia. Additionally, 15 other features showed significant differences among the three groups, and Crohn's disease patients showed an average of almost seven histopathological characteristics. CONCLUSIONS: We described the detailed morphologies of granulomatous lymphangitis on the small bowel mucosa and recommend it as a useful histopathological feature for the diagnosis of Crohn's disease. In terms of specificity and sensitivity, it was superior to non-caseating epithelioid granuloma.

Comparison of intestinal metaplasia in gastric cardia and Barrett's esophagus.

OBJECTIVE: To evaluate and compare the pathological features and immunostaining pattern (cytokeratin 7 (CK-7), mucin core peptide 1 (Muc-1)) in Barrett's esophagus (BE) and cardiac intestinal metaplasia (CIM). METHODS: According to endoscopic diagnosis, patients with gastric cardiac inflammation and BE were selected from March 2008 to February 2009 in Renji Hospital, Shanghai Jiaotong University School of Medicine. Those patients who had histological findings of intestinal metaplasia (82 cases of CIM and 64 special type BE) were enrolled in our study. Hematoxylin-eosin, periodic acid-Schiff and Alcian blue staining and an immunohistochemical examination (CK-7, Muc-1) were undertaken in all of them. RESULTS: Squamous mucosa overlying the columnar crypts with intestinal metaplasia, also called buried metaplasia, was often found in the BE group (56.2%), mainly as an incomplete type (85.9%). Inflammation in the gastric antrum was more severe in the CIM group (45.1% vs 26.6%), in contrast, esophagitis was more severe in the BE group (53.1% vs 35.4%). CK-7 was highly expressed in the BE group (84.4%) in contrast to the CIM group (37.8%). There was no difference in the expression of Muc-1 in these two kinds of intestinal metaplasia (14.1% vs 19.5%). CONCLUSIONS: Buried intestinal metaplasia, mainly as an incomplete type, is the major predominant type of BE. The degree of inflammation in the gastric antrum and esophagus can differentiate BE from CIM to some extent. CK-7 immunohistochemical staining can help identify BE and CIM but Muc-1 cannot.

XRCC1 downregulated through promoter hypermethylation is involved in human gastric carcinogenesis.

OBJECTIVE: To analyze the expression and aberrant methylation of X-ray repair cross-complementing gene 1 (XRCC1) in gastric carcinogenesis, and identify the molecular mechanism of gastric carcinogenesis. METHODS: The method based on methyl binding domain protein (MBD) immuno-precipitation and promoter microarray was employed to screen the gastric cancer-related methylation-sensitive gene. An immunohistochemistry assay was applied to detect the protein expression of XRCC1 in the multistep progression of gastric carcinogenesis. The mRNA expression of XRCC1 was determined by real-time PCR in tumor tissues and their corresponding non-tumorous tissues. The methylation status and Arg194Trp and Arg399Gln polymorphisms of XRCC1 in gastric cancer and gastritis tissues were analyzed by methylation-specific PCR, bisulfite genomic sequencing and direct DNA sequencing, respectively. RESULTS: Promoter microarray screening and identification suggested that XRCC1 was a methylation-sensitive gene. Immunochemistry results showed that XRCC1 protein expression gradually decreased with progression of gastric mucosal lesions (P < 0.05). The positive rate of XRCC1 in patients with well/moderately differentiated gastric cancer was significantly higher than patients with poorly differentiated gastric cancer (P < 0.05). The mRNA expression of XRCC1 in gastric cancer tissues was significantly lower than that in the non-tumorous tissues (P < 0.05). Meanwhile, XRCC1 methylation in gastric cancer tissues was more frequent than that in the gastritis tissues (P < 0.05), and the downregulation of XRCC1 expression was relevant to methylation (P < 0.05). CONCLUSION: The expression of XRCC1 is downregulated in gastric carcinogenesis, and promoter hypermethylation may be one of the mechanisms contributing to its downregulation.

Expression of polycomb protein EZH2 in multi-stage tissues of gastric carcinogenesis.

OBJECTIVE: To study the role and significance of the polycomb group (PcG) protein EZH2 (enhancer of zeste homolog 2) in the multi-step process of intestinal-type gastric carcinogenesis. METHODS: Gastric specimens were obtained from 142 patients with gastric disease, including 34 with chronic non-atrophic gastritis (NCAG), 33 chronic atrophic gastritis (CAG) with intestinal metaplasia (IM), 40 CAG with dysplasia (DYS) and 35 with intestinal-type gastric carcinomas (GC), and 32 Helicobacter pylori-negative controls. The EZH2 protein was stained by the immunohistochemical method and was expressed as the intensity and percentage of the total number of epithelial cells. The chronic gastritis and the grading of dysplasia were classified according to Chinese National Consensus on chronic gastritis and the Padova international classification. RESULTS: The EZH2 protein levels in the specimens of normal gastric tissue, NCAG, CAG with IM, DYS and intestinal-type GC were gradually increased (P < 0.05), but statistical significance was not found between the groups of DYS and GC. CONCLUSION: PcG protein EZH2 plays an important role in the multi-step process of intestinal-type gastric carcinogenesis.

Role of Helicobacter pylori infection on neuronal expression in the stomach and spinal cord of a murine model.

OBJECTIVE: To investigate the effect of Helicobacter pylori (H. pylori) infection on neuronal expressions in the stomach and spinal cord of mice so as to explain dyspepsia symptoms in H. pylori infected patients. METHODS: C57BL/6 female mice were studied at 2 weeks (acute infection group) and 12 weeks (chronic infection group) after H. pylori inoculation. Histological analyses for gastric inflammation and bacterial colonization were assessed by HE staining and Warthin-Starry staining. Fos, vasoactive intestinal polypeptide (VIP) and calcitonin gene-related peptide expressions (CGRP) were studied by immunohistochemistry. RESULTS: H. pylori colonization was present mainly in pyloric region, but bacterial density was similar in both infected groups. The intensity of mucosal inflammation and activity was significantly higher in two infected groups than in those in the control group. The degree of mononuclear and polymorphonuclear cell infiltration in proventricular-glandular region and gastric corpus at 12 weeks after H. pylori inoculation was higher than that at 2 weeks after inoculation. The neuronal expressions of fos, VIP, and CGRP in the stomach and spinal cord were significantly more marked in the infected groups than in the control group, but there was no significant difference between two infected groups. CONCLUSION: H. pylori infection induced different degrees of gastric mucosal inflammation in the murine model. Both early and chronic infection groups of mice showed enhanced neuronal expressions of fos, VIP and CGRP of stomach and spinal cord and these could form a basis for appearance of functional dyspeptic symptoms in patients with H. pylori infection.

[Clinical features of 107 autoimmune hepatitis patients and 30 of them with AIH-primary biliary cirrhosis overlap syndrome].

OBJECTIVE: In order to provide a reliable basis for the diagnosis and treatment of autoimmune hepatitis (AIH) and its overlap syndrome, we investigated the clinical, immunological characteristics of and the therapeutic methods for AIH and AIH-primary biliary cirrhosis (PBC) overlap syndrome. METHODS: One hundred seven patients (77 with AIH and 30 with AIH-PBC overlap syndrome) were enrolled in the study. Their clinical manifestations, serum liver function tests (LFTs) findings, serum immunoglobulins, liver histopathological changes and their responsiveness to the therapies were investigated. RESULTS: The age distribution of AIH patients showed a single peak during their fifties and their main clinical manifestations were malaise, abdominal distension, anorexia and jaundice. Serum gamma globulin and IgG were significantly higher than their normal levels. 74% of the patients were positive for anti-nuclear antibody (ANA), 32% of the patients were positive for anti-smooth muscle antibody (AMA), and over 50% of the patients suffered from concurrent extrahepatic autoimmune diseases. The main histological changes in the liver biopsies were interface hepatitis (65%), lobular hepatitis and rosette formation of liver cells. Bridging necrosis was observed in severe AIH cases. In the AIH-PBC overlap syndrome patients, the levels of serum ALT, AST, GGT, ALP and incidences of ANA and AMA/AMA-M2 were all significantly higher than those of the AIH group. After treating AIH patients with prednisolone and azathioprine (Aza), complete response was seen in 42 cases (70%), sustained response was seen in 26 cases (43%). Sixteen cases had relapses after the withdrawal of the treatment or prednisolone dosage was reduced lower than 10 mg/d. The cases having normal serum ALT, AST, gamma-globulin and IgG levels after treatment were still responding to the reduced prednisolone dosage of 5-10 mg/d without azathioprine added. After combination with ursodeoxycholic acid (UDCA) treatment, the liver function tests (AST, ALT, TBil) of AIH-PBC overlap syndrome patients also significantly improved compared to those before the treatment (P<0.01). CONCLUSION: AIH and AIH-PBC overlap syndrome are not rare in our clinics. Their diagnoses should be based on the clinical presentations, biochemical and immunological indices and liver histological changes. In AIH cases, once their AST, ALT, gamma-globulin and IgG levels return to normal, the prednisolone dosage can be maintained at 5-10 mg/d and Aza can even be withdrawn. Good improvement for patients with AIH-PBC overlap syndrome can be obtained with UDCA and immunosuppression treatment.

The expression and clinical significance of DNA methyltransferase proteins in human gastric cancer.

This study investigates the varied expression of DNA methyltransferase (DNMT) proteins in gastric cancer (GC) and their relationship with the biological behavior of the tissues. Immunohistochemistry was used to detect the expression of the 3 DNMTs in gastric tissues. We discovered that the positive rates of DNMT1, DNMT3a, and DNMT3b expression in GC tissues were 81.6%, 81.6%, and 68.4%, respectively, and they were significantly higher than those of both para-cancerous (39.5%, 50%, and 44.7%) and normal tissues (10.5%, 10.5%, and 7.9%). DNMT1 was well distributed in the cytoplasm and nuclei of tumor cells or glands, while DNMT3a and 3b were well distributed only in the cytoplasm, as shown by staining a dark brown color. A significant correlation between the DNMT1 and DNMT3a proteins (P < 0.01), a low correlation between DNMT3a and DNMT3b (P < 0.05), and no correlation between DNMT1 and DNMT3b (P > 0.05) were observed. DNMT1 protein expression exhibited no correlation with age, lymphnode metastasis, and also tumor differentiation, but it may have had a correlation with gender. The DNMT3 family was not associated with these factors. Therefore, DNMT overexpression is involved in gastric tumorigenesis, but there is no correlation between the DNMTs and the biological behaviors of tissues.

Chemopreventive effects of rofecoxib and folic acid on gastric carcinogenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine in rats.

OBJECTIVES: Epidemiological and experimental studies indicate that non-steroidal anti-inflammatory drugs (NSAIDs) are chemopreventive agents of gastrointestinal cancers, but few studies on gastric cancer have been carried out. A decrease in folic acid supplement and subsequent DNA hypomethylation are related to gastrointestinal cancers, and it has been shown that high-dose folic acid may interfere with gastric carcinogenesis in dogs. The objective of this study was to investigate the effects of rofecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, and folic acid on the chemoprevention of gastric cancer induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in Wistar rats, and to evaluate the cell proliferation of gastric mucosa in different experimental groups. METHODS: Eighty male Wistar rats were randomly divided into five groups (16 rats in each group). In the control group, the rats were given pure water and basal diet. In the MNNG group, the rats received MNNG in drinking water (100 mg/L) and basal diet. In the MNNG + low-dose rofecoxib group, the rats were given MNNG and rofecoxib 5 mg/kg per day with basal diet. In the MNNG + high-dose rofecoxib group, the rats were given MNNG and rofecoxib 15 mg/kg per day with basal diet. In the MNNG + folic acid group, the rats were given MNNG and folic acid 5 mg/kg per day with basal diet. The experiment was terminated at 50 weeks, and all rats were killed. Blood samples of 3 mL were obtained for measurement of serum folic acid concentrations in the control group, the MNNG group and the MNNG + folic acid group by using chemiluminescent method. The stomach was removed from all rats for histopathological examination and immunohistochemical study. Proliferating cell nuclear antigen (PCNA) expression in gastric epithelial cells was also determined. RESULTS: In the MNNG group, five of 11 rats (45.5%) developed gastric cancer, while in all other four groups no gastric cancer was found (P < 0.05). The positivity rate of PCNA expression in the cancerous tissues was significantly higher than that in the non-cancerous tissues (80.0%vs 14.1%, P < 0.05). The positivity rate of PCNA expression in the gastric mucosal cells of the MNNG group was significantly higher than that in the other four groups. The mean serum folic acid concentration of rats was significantly higher in the MNNG + folic acid group (193.70 +/- 60.73 ng/mL) than those in the control group (84.21 +/- 25.26 ng/mL) and the MNNG group (72.27 +/- 16.70 ng/mL, P < 0.05). It was shown that both low- and high-dose rofecoxib as well as folic acid interfered with the development of gastric cancer induced by MNNG in Wistar rats. CONCLUSIONS: The results indicate that rofecoxib as well as folic acid interferes with gastric carcinogenesis induced by MNNG in Wistar rats, and the suppression of gastric cell proliferation may play a crucial role in the chemoprevention of gastric cancer by rofecoxib and folic acid. The higher serum folic acid concentration of rats may play an important role in the prevention of gastric cancer.

[Effects of Helicobacter pylori infection on neural expression of stomach and spinal cord: an experiment with mice].

OBJECTIVE: To investigate the effects of Helicobacter pylori (Hp) infection on neural expression in stomach and spinal cord, and to investigate the mechanism of functional dyspepsia after Hp infection. METHODS: Thirty-five female C57BL/6 mice were randomly divided into three groups: Group A (acute infection group, undergoing intragastric gavage of Hp suspension every other day for 3 times and then observed for 2 weeks, 15 mice), Group B (chronic infection group, undergoing intragastric gavage of Hp suspension every other day for 3 times and then observed for 2 weeks, 15 mice) and control group (undergoing intragastric gavage of normal saline every other day for 3 times and then observed for 2 weeks, 5 mice). After the observation the mice were killed and their stomachs were taken out to undergo gastric histology and bacterial colonization by HE staining and Warthin-Starry staining respectively. Their spinal cords of thoracic and lumbar segments were taken out too. Immunohistochemistry was used to detect the expression of Fos, vasoactive intestinal polypeptide (VIP), and calcitonin gene-related peptide (CGRP) in the stomach and spinal cord. RESULTS: Three mice died 12 weeks after Hp infection. The rate of Hp colonization, mainly localized in pyloric gland region, was greater in Group B than in Group A, and was 0 in the control group. The severity of inflammation as shown by mononuclear cell infiltration, and activity of inflammation as shown by polymorphonuclear cell infiltration, in the pyloric gland region, proventriculus-glandular stomach region, and corpus gland region were more pronounced in Groups A and B, especially in Group B, than in the control group. The expression values of Fos, VIP, and CGRP in the stomach of Group A were 3.1 +/- 1.4, 4.5 +/- 1.8, and 2.4 +/- 0.8 respectively, all not significantly different from those of Group B (3.1 +/- 1.3, 3.5 +/- 1.6, and 2.2 +/- 0.8, all P > 0.05). The expression values of Fos, VIP, and CGRP in the spinal cord of Group A were 3.8 +/- 1.2, 3.2 +/- 1.5, and 2.2 +/- 0.6, all not significantly different from those of Group B (3.4 +/- 0.7, 2.6 +/- 1.2, and 2.5 +/- 1.1, all P > 0.05 for all). However, the neural expression values in both acute and chronic infection groups were significantly higher than those in the control group (2.4 +/- 0.9, 1.6 +/- 0.9, and 1.2 +/- 0.8 in stomach; and 2.0 +/- 1.6, 1.2 +/- 1.1, and 1.2 +/- 1.1 in spinal cord, P < 0.05 for all). CONCLUSION: Hp infection, both acute and chronic, induces gastric histological changes such as inflammation and activity, and enhances the Fos, VIP, and CGRP expression in stomach and spinal cord, which can be a basis for symptom generation in dyspeptic patients with Hp infection.

Gastric distention enhances FOS and calcitonin gene-related peptide expression in the spinal cord and brain of rats.

OBJECTIVE: The purpose of this study was to determine the pathway and mode of transmission of visceral stimuli by investigating the distribution of the FOS and calcitonin gene-related peptide (CGRP) proteins in the central nervous system. METHODS: Twenty-four Sprague-Dawley rats were divided into three groups: study group (n = 12), sham control group (n = 6), and normal control group (n = 6). A balloon was implanted into the stomach of the rats in the study and sham control groups. After 48 h, the rats in the study group had the stomach distended (80 mmHg) for 2 h, after which they were killed and the antrum, thoracic spinal cord and brain were isolated or dissected. The expression of Fos and CGRP in these tissues was detected immunohistochemically. RESULTS: FOS expression in the dorsal horn of the spinal cord, dorsal nucleus of the vagal nerve, nucleus of the solitary tract in the study rats was significantly higher than in the sham and normal controls. However, no difference was found between the three groups in FOS expression in the myenteric plexus. Similarly, gastric distention enhanced CGRP expression significantly in the spinal cord and medulla oblongata and correlated closely with FOS expression in these two areas. CONCLUSIONS: Gastric distention can activate the limbic system, and CGRP plays an important role in the input of visceral stimuli.

[Effect of T-cell vaccination in murine experimental autoimmune hepatitis].

OBJECTIVE: To investigate the effect of T-cell vaccination in murine experimental autoimmune hepatitis (EAH). METHODS: To induce the EAH model, the syngeneic S-100 antigen emulsified in complete Freud's adjuvant was injected intraperitoneally to C57Bl/6 at day 1 and day 7. For T-cell vaccination, splenocytes were removed from animal 2 weeks after induction of EAH and from control animals, and activated in vitro by mitogen stimulation with Concanavalin A (Con A), then inactivated by mitomycin and injected at 5 10(7) cells per animal as T-cell vaccination at 14 and 7 days before first induction of EAH. RESULTS: The histological grade and serum ALT level of the mice who received T-cell vaccination were decrease significantly, compared with that of model group (1.44+/-0.88 vs. 2.33+/-0.87, t=2.24, P<0.05; 63.0U/L+/-23.4U/L vs. 115.0U/L1+/-39.6U/L, t=2.37, P<0.01, respectively); there was no significant change in mice who received irrelevant T-cell vaccination. CONCLUSION: T-cell vaccination with T cells from EAH animals, but not with irrelevant T cells, was able to protect animals from EAH.

[Effects of Chinese herbal compound on the expression of hepatocyte cytochrome P450IIE1 in rats with alcoholic fatty liver].

OBJECTIVE: To study the effect of Chinese herbal compound (CHC) on the expression of hepatocyte cytochrome P450IIE1 in rat model of alcoholic fatty liver (AFL). METHODS: The AFL rats models were established by administering the drinking water with 40%(v/v) ethanol, and the changes of pathology in liver and hepatocyte P450IIE1 expression, as well as the contents of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), vitamin E (VitE) in liver were detected and compared with those in the control group. RESULTS: Fatty degeneration in liver recovered normally in the CHC-treated group. Immunohistochemical and in situ hybridization examination showed that CHC could inhibit the hepatocyte cytochrome P450IIE1 expression markedly, and restore the contents of MDA, SOD, GSH, VitE to nearly normal range. CONCLUSION: CHC can prevent AFL through inhibiting the hepatocyte cytochrome P450IIE1 expression markedly

Significance of cyclooxygenase-2 expression in human primary hepatocellular carcinoma.

AIM: To clarify the significance of cyclooxygenase-2 (COX-2) expression in human primary hepatocellular carcinoma (HCC) and adjacent nontumorous tissues. METHODS: The COX-2 protein and mRNA were investigated in 27 HCC tissues with adjacent nontumorous tissues, and 5 histologically normal liver tissues,using immunohistochemistry and in situ hybridization. RESULTS: The well-differentiated HCC expressed COX-2 protein (5.68+/-1.19) more strongly than moderated HCC (3.43+/-1.98) and poor differentiated HCC (3.33+/-1.50) (P<0.05 respectively), adjacent nontumorous tissues (4.93+/-1.05) and normal liver tissues (3.20+/-1.92) (P<0.01 respectively); More intensive staining of COX-2 in adjacent nontumorous tissues was observed than that in normal liver tissues (P<0.05). There was no significant difference among adjacent nontumorous tissues, moderately differentiated HCC and poorly differentiated HCC (P>0.05). The expression of COX-2 mRNA was observed in the cytoplasm of the cells of HCC and of the hepatocytes in adjacent nontumorous tissues in which COX-2 protein was positive. CONCLUSION: The overexpression of COX-2 in well-differentiated HCC suggests that COX-2 may play a role in the early stages of hepatocarcinogensis.

[Prophylactic and therapeutic effect of oxymatrine on D-galactosamine-induced rat liver fibrosis].

OBJECTIVE: To investigate the prophylactic and therapeutic effect of oxymatrine on experimental liver fibrosis and to reveal its mechanism. METHODS: By establishing D-galactosamine-induced rat liver fibrosis model, we observed the effect of oxymatrine on serum and tissue biochemical indexes, content of liver hydroxyline, expression of TGF?1 mRNA and changes of tissue pathology. RESULTS: There was a decline of liver hydroxyline and serum AST and ALT in oxymatrine group compared to those of the D-GalN group. The hydroxyline content in oxymatrine pretreatment group was (0.50 0.11)mug/mg compared with (0.99 0.14)mug/mg in D-GalN group (t=8.366, P<0.01). The content in oxymatrine treatment group was (0.44 0.04)mug/mg compared with 0.70 0.06 in D-GalN group (t=9.839, P<0.01). The SOD activity was (149.81 15.28) NU/mg in oxymatrine pretreatment group and (95.22 16.33) NU/mg in the model group (t=7.309, P<0.01); (157.68 19.54) NU/mg in the treatment group compared with (119.88 14.94) NU/mg in the model group (t=4.348, P<0.01). MDA in the pretreatment group was (2.06 0.17) nmol/mg, lower than (4.57 0.37) nmol/mg in the model group (t=17.529, P<0.01). In the treatment group, it was (1.76 0.24)nmol/mg, lower than (3.10 0.17) nmol/mg in the model group (t=12.697, P<0.01). TGF?1 mRNA reduced in the pretreatment and treatment groups as compared with that in the model group (0.21 0.01 vs 0.50 0.01, t=48.665, P<0.01; 0.18 0.02 vs 0.38 0.01, t=22.464, P<0.01). Electron microscopy showed that oxymatrine group had milder hepatocyte degeneration and less fibrosis accumulation than did the model group. Microscopy revealed wide septa expansion from the portal area to the central venous, piecemeal and confluent necrosis and pseudo-nodular formation in part of the lobular in the model group. While in oxymatrine group these lesions were much improved. CONCLUSIONS: Oxymatrine shows prophylactic and therapeutic effect in D-galactosamine induced rat liver fibrosis. This is partly by protecting hepatocyte and suppressing fibrosis accumulation through anti-lipoperoxidation.

Multiple genetic alterations and behavior of cellular biology in gastric cancer and other gastric mucosal lesions:H.pylori infection, histological types and staging.

AIM:To investigate the expression of multiple genes and the behavior of cellular biology in gastric cancer (GC) and other gastric mucosal lesions and their relations to Helicobacter pylori (H. pylori) infection, tumor staging and histological subtypes.METHODS:Three hundred and twenty seven specimens of gastric mucosa obtained via endoscopy or surgical resection, and ABC immunohistochemical staining were used to detect the expression of p53, p16, Bcl-2 and COX-2 proteins.H. pylori was determined by rapid urea test combined with patholo-gical staining or 14 Curea breath test. Cellular image analysis was performed in 66 patients with intestinal metaplasia (IM) and/or dysplasia (Dys). In 30 of them, both cancer and the paracancerous tissues were obtained at the time of surgery. Histolo-gical pattern, tumor staging, lymph node metastasis, grading of differentiation and other clinical data were studied in the medical records.RESULTS:p16 expression of IM or Dys was significantly lower in positive H. pylori chronic atrophic gastritis (CAG) than those with negative H. pylori (CAG: 54.8% vs 88.0%, IM:34.4% vs 69.6%, Dys: 23.8% vs 53.6%, all P < 0.05), Bcl-2 or COX-2 expression of IM or Dys in positive H. pylori cases was signi-ficantly higher than that without H. pylori (Bcl-2: 68.8% vs 23.9%, 90.5% vs 60.7%; COX-2: 50.0% vs 10.8%, 61.8% vs 17.8%; all P <0.05). The mean number of most parame-ters of cellular image analysis in positive H. pylori group was significantly higher than that in negative H. pylori group (Ellipser: 53 plus minus 14, 40 plus minus 12&mgr;m, Area(1): 748 plus minus 572, 302 plus minus 202&mgr;m(2), Area(2): 3050 plus minus 1661, 1681 plus minus 1990&mgr;m(2), all P< 0.05; Ellipseb: 79 plus minus 23, 58 plus minus 15&mgr;m, Ratio-1: 22% plus minus5%,13% plus minus4%,Ratio-2:79% plus minus17%,53% plus minus20%,all P<0.01). There was significant correl-ation between Bcl-2 and histologic pattern of gastric carcinoma, and between COX-2 and tumor staging or lymph node metasta sis (Bcl-2: 75.0% vs16.7%; COX-2: 76.0% vs 20.0%, 79.2% vs 16.7%; all P< 0.05).CONCLUSION:p16, Bcl-2, and COX-2 but not p53 gene may play a role in the early genesis/progression of gastric carcinoma and are associated with H. pylori infection. p53 gene is relatively late event in gastric tumorigenesis and mainly relates to its progression. There is more cellular-biological behavior of malignant tumor in gastric mucosal lesions with H. pylori infec-tion. Aberrant Bcl-2 protein expression appears to be preferentially associated with the intestinal type cancer. COX-2 seems to be related to tumor staging and lymph node metastasis.