[A comparison of efficacies between transjugular intrahepatic portosystemic shunt versus portoazygos devascularization in the treatment of portal hypertension with variceal bleeding].

OBJECTIVE: To compare the efficacies between transjugular intrahepatic portosystemic shunt (TIPS) and portoazygos devascularization (PAD) in the treatment of portal hypertension with variceal bleeding. METHODS: From December 1993 to December 2010, 309 patients with portal hypertension and variceal bleeding were admitted. According to their general conditions and Child-Pugh grades, they were assigned to undergo TIPS (group A, n = 235) or PAD (group B, n = 74). Before operation, compared with the PAD group, the TIPS group possessed worse liver functions, more severe ascites and a greater frequency of bleeding. After operation, the therapeutic efficacies and changes of portal hemodynamics, recurrent variceal bleeding, post-operative encephalopathy and long-term survival were evaluated between two groups. RESULTS: The postoperative portal pressure in the TIPS group ((42.6 ± 7.0) vs (26.3 ± 4.1) cm H2O) decreased much more than that in the PAD group ((38.7 ± 5.2) vs (33.5 ± 5.8) cm H2O, P < 0.01). The rebleeding rates during early postoperation were 0.85% (2/235) and 6.76% (5/74) in TIPS and PAD groups respectively, the occurring rates of hepatic encephalopathy 4.68% (11/235) and 4.05% (3/74) and the rates of operative mortality 1.70% (4/235) and 6.76% (5/74) respectively. Survival rates of 1, 3, 5 and 10 years were 98.30% (231/235) vs 92.24% (69/74), 92.41% (146/158) vs 88.06% (59/67), 80.77% (84/104) vs 79.25% (42/53), 51.43% (36/79) vs 51.85% (14/27) in TIPS and PAD groups respectively. CONCLUSIONS: As compared with PAD, TIPS offers the such advantages as less trauma, wider indication, faster hemostasis and satisfactory therapeutic efficacies. Especially for the emergency treatment of a patient with massive variceal bleeding and Child-Pugh C grade liver function, TIPS is a better option than PAD.

[Observation on experimental liver fibrosis and hepatic carcinogenesis of HBV gene knock-in transgenic mice induced by CCl4/ethanol].

OBJECTIVE: To study the effects of carbon tetrachloride (CCl4)/ethanol induction upon experimental liver fibrosis and hepatic carcinogenesis of HBV transgenic mice. METHODS: The wild-type mice, p21-HBx transgenic mice with integration of p21 locus by HBx gene and p21-HBsAg transgenic mice with integration of p21 locus by HBsAg gene were induced separately by CCl4/ethanol twice weekly for 20 weeks. The investigators observed the development of liver fibrosis and hepatic carcinogenesis in three groups and detected the gene expressions of HBx and HBsAg by RT-PCR. RESULTS: The expression of HBx or HBsAg mRNA existed in both control and induced transgenic mice, but in none of wild-type mice. Comparing with wild-type mice, p21 genes was not expressed in livers of transgenic mice. After induction by CCl4/ethanol, the fibrotic degrees of liver were not significantly different among wild-type mice, p21-HBx transgenic mice and p21-HBsAg transgenic mice, as well as between male and female mice. Reversely, the incidence rates of hepatic carcinogenesis of two HBV gene knock-in transgenic mouse lines (p21-HBx & p21-HBsAg) were higher than that of wild-type mice. And the incidence rate of hepatic carcinogenesis in males was also markedly higher than that in females. Induction by CCl4/ethanol markedly promoted and accelerated hepatic carcinogenesis in transgenic mice. CONCLUSIONS: Integration of HBsAg and HBx genes into the murine p21 locus can significantly promote the progression of hepatic carcinogenesis, but failed to promote the progression of liver fibrosis. The male mouse is more likely to develop experimental hepatocellular carcinoma than the female mouse. Experimental hepatocellular carcinoma induced by CCl4/ethanol in p21-HBx and p21-HBsAg transgenic mice is a feasible animal model.

[Effects of Smad4 on liver fibrosis and hepatocarcinogenesis in mice treated with CCl4/ethanol].

To study the effects of Smad4 on liver fibrosis and hepatocarcinogenesis in mice treated with CCl(4)/ethanol. The wild-type mice (Smad4 +/+) and the Smad4 knockout mice (Smad4 +/-) were injected subcutaneously with carbon tetrachloride(CCl(4))/ethanol twice a week for twenty weeks. The expression of Smad4, TGFbeta1, Smad2, Smad3, Smad6, TIMP1, MMP2 and MMP9 was detected by RT-PCR. In the cirrhotic liver, the expression of Smad4 mRNA was significantly higher than that in the normal liver. Comparing with wild-type mice (Smad4 +/+), the TGFbeta1-Smad4 signaling was markedly attenuated in the Smad4 knockout mice (Smad4 +/-). After induction by CCl(4)/ethanol, the hepatic fibrosis in the Smad4 knockout mice (Smad4 +/-) was obviously alleviated compared with the wild-type mice (Smad4 +/+), and the incidence rate of hepatocarcinogenesis of the former was also lower than that of the latter(32.0% vs 41.9%). These results indicate that knocking out Smad4 can delay the progression of liver fibrosis and liver cancer.

Clinical analysis of surgical treatment of portal hypertension.

AIM: To review the experience in surgery for 508 patients with portal hypertension and to explore the selection of reasonable operation under different conditions. METHODS: The data of 508 patients with portal hypertension treated surgically in 1991-2001 in our centers were analyzed. Of the 508 patients, 256 were treated with portaazygous devascularization (PAD), 167 with portasystemic shunt (PSS), 62 with selective shunt (SS), 11 with combined portasystemic shunt and portaazygous devascularization (PSS+PAD), 9 with liver transplantation (LT), 3 with union operation for hepatic carcinoma and portal hypertension (HCC+PH). RESULTS: In the 167 patients treated with PSS, free portal pressure (FPP) was significantly higher in the patients with a longer diameter of the anastomotic stoma than in those with a shorter diameter before the operation (P < 0.01). After the operation, FPP in the former patients markedly decreased compared to the latter ones (P < 0.01). The incidence rate of hemorrhage in patients treated with PAD, PSS, SS, PSS+PAD, and HCC+PH was 21.09% (54/256), 13.77 (23/167), 11.29 (7/62), 36.36% (4/11), and 100% (3/3), respectively. The incidence rate of hepatic encephalopathy was 3.91% (10/256), 9.58% (16/167), 4.84% (3/62), 9.09% (1/11), and 100% (3/3), respectively while the operative mortality was 5.49% (15/256), 4.22% (7/167), 4.84% (3/62), 9.09% (1/11), and 66.67% (2/3) respectively. The operative mortality of liver transplantation was 22.22% (2/9). CONCLUSION: Five kinds of operation in surgical treatment of portal hypertension have their advantages and disadvantages. Therefore, the selection of operation should be based on the actual needs of the patients.

[Effects of portaazygous disconnection, portocaval shunt and selective shunts on experimental rat liver cirrhosis].

OBJECTIVE: To evaluate the effects of portaazygous disconnection (PAD), portacaval shunt (PCS) and distal splenocaval shunt (DSCS) on the portosytemic shunting (PSS), hepatic function (HF), hepatic mitochondrial respiratory function (HMRF), oral glucose tolerance test (OGTT) and arterial ketone body ratio (KBR) in order to provide a sound basis for selecting suitable operations for patients. METHODS: Using a cirrhotic portal hypertensive model induced by CCl4/ethanol in Wistar rats, the PSS, HF, HMRF, OGTT and KBR were determined three weeks after PCS, DSCS and PAD. RESULTS: It was revealed that: (1) In the cirrhotic portal hypertension rats, the PSS increased significantly, HMRF and hepatic reserve function (HRF) decreased significantly when compared with the control rats. (2) At the time of first postoperative week, the mean blood glucose value in the 120-minute OGTT in each PAD, PCS and DSCS groups had significant differences compared with the cirrhotic control group. But during the second and third postoperative weeks, the mean blood glucose values in the 120-minute OGTT in both PAD and DSCS groups had no significant differences compared with the cirrhotic control group except for the PCS group. The values of KBR in the three operative groups decreased significantly compared with the cirrhotic control group during the two postoperative weeks. In the third postoperative week, only the values of KBR in the PCS group had a significant difference compared with the cirrhotic control group. (3) After PCS, the PSS was further increased; HF and HMRF were significantly decreased. Little improvement was found in the third postoperative week. (4) After DSCS and PAD, the above mentioned indices were less influenced, and they were restored more quickly than those in the PCS group. CONCLUSION: We found that PAD and DSCS are more desirable than PCS.

Inhibitory effect of retroviral vector containing anti-sense Smad4 gene on Ito cell line, LI90.

BACKGROUND: Transforming growth factor-beta1 (TGF-beta1) exerts strong fibrogenic potential in culture-activated HSCs. Smad4 is a key intracellular mediator for the transforming growth factor-beta (TGF-beta) superfamily of growth factors. The aim of this study was to assess the effects of the antisense Smad4 gene on Ito cell line, LI90. METHODS: The recombinant retroviral vector pLXSN-Smad4 was constructed by cloning the rat antisense Smad4 cDNA into the retroviral vector pLXSN. Retroviruses with or without the antisense gene were obtained by transfecting pLXSN-Smad4 and pLXSN vectors into PA317 cells. Human hepatic stellate cells (HSCs) LI90 were infected with these retroviruses followed by selection with G418. The expression of Smad4 was detected by Northern and Western blots. Cell biological characteristics, including cell growth curve, 3H-TdR and 3H-proline uptake by HSCs and the production of extracellular matrix were assessed. RESULTS: mRNA and protein expressions of Smad4 in LI90 cells transfected with retrovirus containing the antisense Smad4 gene were much lower than those in LI90 cells transfected with empty vector or parental LI90 cells. Cells hypoexpressing the Smad4 gene exhibited a slower rate of growth, a lower uptake of 3H-TdR and 3H-proline (P < 0.01), and smaller production of th extracellular matrix, compared with parental LI90 cells and cells transfected with empty retrovirus. CONCLUSIONS: The antisense Smad4 gene can suppress the expression of the Smad4 gene, reduce endogenous production of Smad4 mRNA and protein, block TGF-beta1 signaling pathway, inhibit activation of Ito cells, obstruct the growth of Ito cells, decrease the production of the extracellular matrix (ECM). Our results may provide a basis for the development of antifibrotic gene therapy.

[Obstruction of TGF-beta1 signal transduction can decrease the process of hepatocellular carcinoma in mice induced by CCl4/ethanol].

OBJECTIVE: To study obstruction of the TGF-beta(1) signal transduction by antisense RNA of Smad(4) and its effects on experimental hepatic carcinoma of mice. METHODS: We used the mouse model of primary hepatic carcinoma induced by CCl(4)/ethanol, and transferred antisense Smad(4)cDNA with retrovirus-mediated via portal vein infusion into liver. Southern Blot confirmed that the antisense Smad(4)cDNA had been integrated into the liver. The antisense Smad(4) gene could down-regulate the expression of Smad(4) in fibrotic liver observed by Northern and Western Blot. RESULTS: In the non-therapeutic cirrhotic liver, the expression of Smad(4) mRNA was significantly increased than normal liver. After antisense Smad(4) gene was transferred, the expression of Smad(4) mRNA in the therapeutic liver was significantly decreased compared with non-therapeutic cirrhotic liver. The fibrotic degree of therapeutic liver was alleviated compared with the non-therapeutic fibrotic liver. No significant difference was found between the rates of carcinogenesis of non-therapeutic cirrhotic liver and that of therapeutic cirrhotic liver. But the diameters and numbers of the liver cancers in the therapeutic cirrhotic group were less than that in the non-therapeutic cirrhotic group. CONCLUSION: These results indicate that the antisense Smad(4) gene not only can obstruct the progression of liver fibrosis, but also can inhibit the progression of liver cancer, by obstructing the signal transduction of TGF-beta1.

[Effects of anti-sense Smad4 gene on the biological characteristics of the fat-storing cell line CFSC].

OBJECTIVE: To investigate the effects of antisense Smad(4) on the biological characteristics of the fat-storing cell line CFSC. METHODS: Fat-storing cells of line CFSC from rat with liver fibrosis were cultured and transfected with 50 MOI of recombinant adenoviral vector carrying antisense Smad(4) (AdvATSmad(4)) or the control empty adenovirus (Adv0), both produced by 293 packaging cells, respectively. Two, four, and six days after the transfection the cultured cells were collected to undergo trypan blue staining and cell counting. The growth curves were drawn. The presence of antisense Smad(4) was detected by RT-PCR and Western blotting. (3)H-TdR was added into the culture media to be co-cultured for 6 hours. Then the cells were collected to examine the (3)H-TdR incorporation rate. RT-PCR and immunohistochemistry were used to examine the expression of COL1A1 and type I collagen, kinds of extracellular matrix (ECM). RESULTS: Compared with the control CFSC and the Adv0-transfected CFSC cells, the cell growth curve, (3)H-TdR incorporation rate, proline incorporation rate, expression of Smad(4), and expression of extracellular matrix were markedly decreased in the AdvATSmad(4)-transfected CFSCs. CONCLUSION: The antisense Smad(4) gene inhibits the expression of Smad(4) mRNA and protein, proline incorporation and cell growth, thus down-regulating the production of ECM. Antisense Smad(4) gene may be used as a choice of gene therapy for liver fibrosis.

[Obstruction of TGF-beta1 signal transduction by anti-Smad4 gene can therapy experimental liver fibrosis in the rat].

OBJECTIVE: To study the therapeutic effects to block the TGF-beta1 (transforming growth factor beta1) signal transduction by antisense Smad4 gene on experimental fibrotic liver. METHODS: Using the rat model of liver fibrosis induced by Carbon Tetrachloride (CCl4)/ethanol, we transfected antisense Smad4 gene mediated by adenovirus via portal vein infusion into the liver, and observed the expression of Smad4 by Retro-Polymerase Chain Reaction (RT-PCR) and Western Blot. We also investigated the pathologic features and collagen expression. RESULTS: In the non-therapeutic cirrhotic liver, the expression of Smad4 mRNA was significantly increased than normal liver, and so was the collagen I. After antisense Smad4 gene being transfected, the expression of Smad4 mRNA and that of collagen I in the therapeutic liver was significantly decreased, compared with the non-therapeutic cirrhotic liver. The fibrous degree of therapeutic liver was also reduced compared with the non-therapeutic fibrous liver. CONCLUSION: These results indicate that because antisense Smad4 gene could block TGF-beta1 signal transduction by reducing the expression of Smad4, so it could inhibit the production of extracellular matrix (ECM) and improve hepatic fibrosis.