Differentiation of hepatocytoid cell induced from whole-bone-marrow method isolated rat myeloid mesenchymal stem cells.

AIM: To explore the expansion and differentiation of hepatocytoid cell induced from myeloid mesenchymal stem cell (MSC) in vitro, in order to find suitable resource of hepatocytes for bioartificial liver or liver transplantation. METHODS: The rat myeloid MSC was isolated and divided into three groups which were cultured by Friedensteion method, and then were induced by culture fluid, culture fluid plus cholestatic serum and culture fluid plus hepatocyte growth factor (HGF), respectively. Hepatocytoid cell as well as expression of CK18 and AFP was observed by immunohistochemistry. RESULTS: After the induction for 21 d, hepatocytoid cell was observed, and its expression of CK18 and AFP was detected by immunohistochemistry in MSC cultured with cholestatic serum. Furthermore, on the 35th d, albumin mRNA was expressed in the cell, suggesting the inducing effect was similar to that by HGF. CONCLUSION: Rat myeloid MSC can differentiate into hepatocyte lineage under appropriate condition. This method is easy to operate.

[Study on the effect of heluoshugan capsule on liver fibrosis induced by Schistosoma japonicum infection in mice].

OBJECTIVE: To investigate the effect of Chinese traditional medicine heluoshugan capsule on liver fibrosis induced by Schistosoma japonicum infection in mice. METHODS: Liver fibrosis in mice was established by Schistosoma japonicum infection in 6 weeks. Suspension of heluoshugan prepared with normal saline was given orally to the mice, 2 capsules for 20 mice daily for 8 weeks. The level of vascular endothelial growth factor (VEGF), focal adhesion kinase (FAK) and type I, III collagen in liver tissue were detected by immuno-histochemistry. RESULTS: The results showed that heluoshugan improved the pathological change of the liver tissue, decreased the level of type I, III collagen, especially type III collagen (P < 0.01). The level of VEGF and FAK expression was inhibited after the administration of heluoshugan, though the level usually increased in liver fibrosis due to the infection. CONCLUSIONS: The result suggests that heluoshugan capsule might have therapeutic effect on liver fibrosis induced by the infection of Schistosoma japonicum in mice, by inhibiting the activation of hepatic stellate cells and the pathological change of liver blood vessel.

[In vitro study on Hep G2 cell infected by hepatitis C virus].

OBJECTIVE: To establish hepatitis C virus (HCV) infected cell model which is similar to the infection in vivo and can support HCV to replicate for a long time. METHODS: After infected with HCV-positive serum, Hep G2 cells were cultured for 60 days. Nested RT-PCR was used to detect plus and minus HCV RNA in cultured cells and supernatants. RESULTS: Plus HCV RNA was detected intermittently in Hep G2 cells during 2-30 days, minus HCV RNA was detected during 3-30 days after infection, the detection rate was similar to plus HCV RNA. Plus and minus HCV RNA can be still intermittently detected during 31-60 days after infection. However, the detection rate gradually declined. Plus HCV RNA was also found intermittently positive in the supernatant, and the detection rate was consistent to that in cells. Minus HCV RNA was not detected in the supernatant. CONCLUSION: Hep G2 cells were susceptible to HCV, and could support HCV to replicate for a relatively long time. Hep G2 is an ideal HCV infection cell model.

[Expression of hepatic Bcl-2 and Bax proteins in schistosome-infected mice and the role of pentoxifylline].

OBJECTIVE: To study the expression of hepatic Bcl-2 and Bax proteins in mice infected with Schistosoma japonicum and the role of pentoxifylline (PTX) in the expression. METHODS: Forty mice were randomly divided into 4 groups: one normal control group,mice in the other three groups were all infected each with 25 cercariae, the infected control group was fed for 10 weeks after infection, and 2 weeks after infection, the high dose PTX group was given PTX 360 mg/(kg x d) for 8 weeks and the low dose PTX group was given PTX 180 mg/(kg x d)also for 8 weeks. At the end of 10 weeks all the mice were killed. Bcl-2 and Bax proteins expression was detected by immunohistochemisty. RESULTS: Compared with the normal control group, the expression of Bcl-2 and Bax was significantly higher in the infected control group (P < 0.05). Bcl-2 was significantly higher in high (dose PTX group than in the infected control group and in low dose PTX group (P < 0.05). However there was no significant difference in the expression of Bax among the groups (P > 0.05). CONCLUSION: PTX treatment can significantly increase the expression of Bcl-2 in liver tissue of schistosome-infected mice in a dose-dependent manner, and may play a role against liver inflammation and schistosomiasis-related liver fibrosis.

[Experimental study on enzyme dot assay for detection of hemorrhagic fever with renal syndrome antigen].

OBJECTIVE: To establish a new and efficient method(IEDA) for detection of hemorrhagic fever with renal syndrome virus (HFRSV) antigen. METHODS: An immune enzyme dot assay (IEDA) with mixture of three sorts anti-HFRSV-IgG, which was obtained from rabbit vaccinated with EHFV R22, Chen and Hubei strain was employed to detect HFRSV antigen in serum and urine from epidemic hemorrhagic fever (EHF) patients, and compared with indirect immune fluorescence assay (I-IFA), 76 serum samples and 40 urine samples were detected in this study. RESULTS: The results showed that the total positive rate of HFRSV antigen detected by IEDA was 73.68% in serum and 65.00% in urine, while that detected by I-IFA was 75.00% and 70.00%, respectively. The positive rate in primary phase (within 5 days) of HFRSV antigen detected by IEDA was 94.34% in serum and 83.33% in urine, while that detected by I-IFA was 64.42% and 55.56%, respectively, there was significant difference in both serum and urine detections. Correlation study showed a high correlation in the result of IEDA and I-IFA. CONCLUSION: The results of this study suggested that the IEDA, as compared with I-IFA, was a more specific, sensitive, rapid and simple method with higher positive rate in primary phase. IEDA could be widely used for early diagnosis of HFRS in hospital at grassroots level.

Promoting hepatic growth factor in the treatment of heavy type hepatitis and severe chronic hepatitis: a multicenter clinical study.

BACKGROUND: The mortality rate of heavy type hepatitis is high. No special treatment is available except general treatment. This multicenter clinical study was designed to observe the safety and efficacy of promoting hepatic growth factor (PHGF) in the treatment of heavy type hepatitis and severe chronic hepatitis. METHODS: 347 patients with heavy type hepatitis and 324 with severe chronic hepatitis were subjected to administration of 120 microg of PHGF per day for 4 weeks on the basis of general treatment. Those who were being effectively treated would last additional 2 to 4 weeks. Blood routine, urine routine, blood urea nitrogen (BUN), blood creatinine (Cr), blood ammonia, alpha fetoprotein (AFP), electrolyte, alanine transaminase (ALT), aspartate transaminase (AST), serum total bilirubin (TBIL), serum direct bilirubin (DBIL), prothrombin time activity (PTA), total protein (TP) and albumin (ALB) were detected in the patients before treatment, 2 weeks after treatment, and at the end of the treatment. Any side-effect would be recorded. RESULTS: In the patients with severe chronic hepatitis, the total effective rate of the treatment was 88.9%. The levels of ALT, AST and TBIL decreased significantly (P<0.001), whereas those of PTA and ALB increased significantly (P<0.001), and the level of AFP increased slightly. In patients with heavy type hepatitis, the total effective rate of this treatment was 78.4%, and patients at different stage showed different results. The total effective rates of patients with early, medium and terminal stage heavy type hepatitis were 89.9%, 84.8% and 27.5%, respectively. No severe side-effect was shown. CONCLUSION: PHGF is effective and safe in the treatment of patients with heavy type hepatitis and severe chronic hepatitis. But it should be administered early in patients with heavy type hepatitis so as to get better curative effects.

[Expression of IL-2 and TNF-alpha in the liver and the effect of injection of these cytokines on liver fibrosis in mice infected with Schistosoma japonicum].

OBJECTIVE: To detect the expression of IL-2 and TNF-alpha in the liver at different period postinfection of Schistosoma japonicum and their effect on liver fibrosis after supplementary injection of these cytokines. METHODS: Mice were infected with schistosome cercariae and divided into 3 groups. Two groups were injected (i.p.) every other day with IL-2 and TNF-alpha respectively for consecutive 4 wk. The third group and an uninfected group of normal mice were regarded as control. The ABC immunohistochemistry and pathologic image multimedia quantification system were applied to detect activity of IL-2 and TNF-alpha. RESULTS: The level of IL-2 and TNF-alpha in the liver in infected but untreated group slowly decreased (from 8, 11, 14 to 18 wk). The supplementary injection of the cytokines at 6 wk postinfection in the two groups increased the cytokines significantly, the level of IL-2 or TNF-alpha was higher at 1-8 wk after the last injection than that of both infected and uninfected control groups (P < 0.01). The granulomatous inflammation and fibrosis in the livers of the two groups were slighter than that of the control. CONCLUSION: At the 6th wk postinfection with egg deposition, exogenous supplementation with TNF-alpha or IL-2 induces enhanced expression of the two kinds of cytokines, corresponding to a diminished degree of the liver granulomatous inflammation and fibrosis.

Effects of pentoxifylline on the hepatic content of TGF-beta1 and collagen in Schistosomiasis japonica mice with liver fibrosis.

AIM: To study the effects of pentoxifylline (PTX) on the content of hepatic TGF-beta1, type I and type III collagen in schistosomiasis japonica mice with liver fibrosis and its mechanism of anti-fibrosis. METHODS: Forty mice with schistosomiasis were divided into four groups: one group as control without any treatment, other three were treated with Praziquantel 500 mg/(kg x d)for 2 d, high dose PTX 360 mg/(kg x d) for 8 wk, and low dose PTX 180 mg/(kg x d) for 8 wk respectively. Immunohistochemical technique and multimedia color pathographic analysis system were applied to observe the content change of hepatic TGF-beta1, type I and type III collagen in schistosomiasis japonica mice with liver fibrosis before and after PTX treatment. RESULTS: Effects of PTX on the content change of hepatic TGF-beta1, type I and type III collagen in schistosomiasis japonica mice with liver fibrosis were related to the dosage of PTX, high dose PTX treated group could significantly reduce the content of TGF-beta1 (0.709+/-0.111), type I (0.644+/-0.108) and type III (0.654+/-0.152) collagen compared with those of control group (0.883+/-0.140, 0.771+/-0.156, 0.822+/-0.129) with statistical significance (P<0.05). Low dose PTX could also reduce the hepatic content of TGF-beta1 (0.752+/-0.152), type I (0.733+/-0.117) and type III (0.788+/-0.147) collagen, but without statistical significance (P>0.05). Both high dose and low dose PTX groups have significant differences on the content of TGF-beta1, type I and type III collagen (P<0.05, P<0.05, P<0.01, respectively). CONCLUSION: High dose of PTX treatment could reduce the content of hepatic TGF-beta1, type I and type III collagen significantly in schistosomiasis japonica mice with liver fibrosis, and thus plays its role of antifibrosis.

[Effect of pentoxifylline on the expression of hepatic TGF-beta 1, type I and type III collagen in mice with liver fibrosis due to Schistosoma japonicum infection].

OBJECTIVE: To study the effect of pentoxifylline (PTX) on the content of hepatic TGF-beta 1, type I and type III collagen in schistosome-infected mice with liver fibrosis. METHODS: Forty mice with schistosomiasis were divided into four groups: one group as control without any treatment, other three were treated with praziquantel 500 mg/(kg.d) for 2 d, high dose PTX 360 mg/(kg.d) for 8 wk, and low dose PTX 180 mg/(kg.d) for 8 wk respectively. Immunohistochemical technique and multimedia color pathographic analysis system were applied to observe the content of hepatic TGF-beta 1, type I and type III collagen in mice infected with S. japonicum before and after treatment. RESULTS: The effect of PTX on the content of hepatic TGF-beta 1, type I and type III collagen in mice was related to the dosage of PTX. High dose PTX treatment significantly reduced the content of TGF-beta 1, type I and type III collagen compared to the control (P < 0.01), whereas no difference was found between the group of low dose PTX treatment and control (P > 0.05). CONCLUSION: High dose PTX treatment could reduce the content of hepatic TGF-beta 1, type I and type III collagen significantly in schistosome-infected mice with liver fibrosis.

Intestinal flora translocation and overgrowth in upper gastrointestinal tract induced by hepatic failure.

AIM:To explore the relationship between endoinfection caused by intestinal flora translocation and multiple organ dysfunction in hepatic failure.METHODS:By using the quantitative bacteria culture, bacteria colony was counted in GI tract, bile duct and mesenteric lymphonodus in rat hepatic failure model.RESULTS:Intestinal flora migrated up to the upper GI tract and overgrew in stomach and jejunum in rats with hepatic failure.The number of bacteria colonies in the specimens of stomach, jejunum and ileum were 4.7X10(4)/mL, 2.1X10(5)/mL, 5.5X10(6)/mL in experiment group and 4.6X10(2)/mL, 6.1X10(1)/mL, 2.4X10(3)/mL in control group respectively (P < 0.05 ). Bacteria in bile duct and mesenteric lymphonodus of hepatic failure rats were also cultured. Extensive damages of gastrointestinal mucosa caused by bacterial overgrowth were observed.CONCLUSION:Intestinal flora translocation and overgrowth in stomach and jejunum formed an endoinfectious source and caused obvious pathological injury of gastrointestinal mucosa, which play a very important role in developing abdominal distension, toxic intestinal expansion, alimentary tract haemorrhage and endotoxemia in patients with hepatic failure.