Construction of a hepatic stellate cells subtracted cDNA library of differentially expressed genes in normal mice and mice with Schistosomiasis japonica / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

To construct a hepatic stellate cells (HSCs) subtracted cDNA library to find differentially expressed genes in normal mice and mice infected with Schistosoma japonicum (S. japonicum). Suppression subtractive hybridization (SSH) was used. The cDNA fragments of normal mouse were compared to those of schistosoma-infected mice to find differentially expressed genes. Then differentially expressed cDNA fragments were directly inserted into T/A cloning vector to set up the subtractive library. Amplification of the library was carried out with transformation of DH5alpha. The amplified library contained more than 400 positive bacterial clone, which were then hybridized with forward and backward subtracted probes for differential screening. One hundred positive bacterial clones were randomly selected for sequencing and BLAST analysis. Finally, virtual Northern Blot confirmed such differential expression. The subtracted cDNA library of differentially expressed genes of HSCs was constructed successfully, the library is efficient and lays foundation for screening and cloning new and specific genes of schistosomiasis.

The diagnostic value of eight serum indices for liver fibrosis / 中华肝脏病杂志

<p><b>OBJECTIVES</b>Using receiver operating characteristic (ROC) curve to compare the value of platelet derived growth factor-BB (PDGF-BB), transforming growth factor-beta1 (TGF-beta1), matrix metalloproteinase-1 (MMP-1), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), hyaluronic acid (HA), type III procollagen (PCIII), type IVcollagen (C-IV) and laminin (LN) in serum and message RNA (mRNA) expression of TIMP-1 and MMP-1 in peripheral blood mononucleocytes (PBMC) to diagnose liver fibrosis.</p><p><b>METHODS</b>Serum samples from 60 chronic hepatitis B patients and 20 healthy blood donors were assayed for serum level of PDGF-BB, TGF-beta1, MMP-1 and TIMP-1 with ELISA, and for serum level of HA, PCIII, C-IV and LN, with RIA. The mRNA expression of TIMP-1 and MMP-1 in PBMC was detected by RT-PCR. Liver biopsy was performed in all those patients. The biopsy materials were examined histopathologically.</p><p><b>RESULTS</b>Through the analysis by ROC curve, serum PDGF-BB is the most valuable marker, and its sensitivity was the highest among eight indices. The marker with the highest specificity was TIMP-1 mRNA in PBMCs. The area under the curve (AUC) of PDGF-BB, TIMP-1, HA, PCIII, C-IV, LN, TIMP-1 mRNA was 0.985, 0.726, 0.318, 0.728, 0.727, 0.583, 0.463, 0.876, respectively. The sensitivity and specificity of combination of PDGF-BB and TIMP-1 mRNA were 97.4%, 95.0%, respectively.</p><p><b>CONCLUSION</b>Serum PDGF-BB is the most potential index among eight markers. The combination of serum PDGF-BB and TIMP-1 mRNA in PBMC might be more efficient to screen liver fibrosis.</p>

Expression of transforming growth factor-beta1 in renal fibrosis of human mesengial proliferative glomerulonephritis / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To explore the possible effect of transforming growth factor-beta(1) (TGF -beta(1)) on the development of renal fibrosis in human mesengial proliferative glomerulonephritis (MsPGN).</p><p><b>METHODS</b>Immunohistochemistry method, sirius red staining polarization microscopy and the computer imaging analysis system were used to detect the expression of TGF-beta(1), the distribution of collagen I, collagen III and collagen IV.</p><p><b>RESULT</b>In MsPGN with renal fibrosis, collagen IV was increased markedly,and collagen I and collagen III appeared in the expanded mesengial matrix abnormally. Collagen III and collagen IV were increased markedly in tubulointerstitium. TGF-beta(1) expression was positively correlated with the expression of collagen I, collagen III and collagen IV in tubulointerstitium (r=0.82 0.92,P<0.01), and negatively correlated with I/III, I/IV and III/IV (r=-0.83,-0.92, P<0.001).</p><p><b>CONCLUSION</b>Abnormal increase of TGF-beta(1) may be one of the important factors associated with glomerular sclerosis and tubulointerstitial fibrosis through the increment and abnormal distribution of collagen I, collagen III and collagen IV.</p>

Expression of Bcl-2 and Bax proteins in rat liver fibrosis model and the role of IFN-gamma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the expression of Bcl-2 and Bax in rat liver fibrosis model induced by CCl4 and the role of IFN-gamma.</p><p><b>METHODS</b>Liver fibrosis was induced in rats by subcutaneous injection of CCl4. The rats were divided into fibrosis model group, Bie-Jia-Ruan-Gan-Tablet treatment group and IFN-gamma treatment group (0.2 MU.kg.d, i. m. for 12 weeks), and another 10 rats without any treatment were used as normal control. Bcl-2 and Bax proteins expression were detected by immunohistochemistry.</p><p><b>RESULTS</b>Bcl-2 was expressed weakly in the homogenate of hepatocytes and hepatic sinusoid in normal control rats, and it was expressed stronger in fibrous septae, portal area, hepatic sinusoid, the homogenate and membrane of hepatocytes and central vein in fibrosis model rats (3.87%+/-2.37% vs 9.46%+/-4.29%, t=2.83, P<0.05). Bax was expressed slightly in central vein and the hepatic sinusoid around, and it was expressed stronger in the homogenate of hepatocytes, hepatic sinusoid, fibrous septae, membrane of hepatocytes and epithelial cells of bile duct (3.50%+/-1.88% vs 9.80%+/-3.75%, t=3.72, P<0.01). Compared with that in fibrosis model rats, the expression of Bax was significantly lower in rats treated with IFN-gamma (9.80%+/-3.75% vs 5.85%+/-2.35%, t=2.98, P<0.01), but the expression of Bcl-2 was not significantly different (t=1.49, P>0.05), however it was significantly lower in fibrous septae in IFN-gamma-treated group than in model group (6.58%+/-4.13% vs 9.46%+/-4.29%, t=2.80, P<0.05).</p><p><b>CONCLUSION</b>The expression of Bcl-2 and Bax increases in liver fibrosis model rats, and IFN-gamma can promote myofibroblasts apoptosis</p>

Study on the influence factors of the serum fibrosis markers / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To analyse the factors which influence the four serum fibrosis markers hyaluronic acid (HA), type III procollagen (PCIII), laminin (LN) and type IV collagen (CIV).</p><p><b>METHODS</b>The levels of serum HA, PCIII, LN and CIV were measured by RIA in 141 patients with chronic hepatitis B (CHB), then the patients were divided into two groups according to the serum fibrosis markers, namely consistent group and inconsistent group. the liver biopsy materials were examined pathomorphologically and liver function was detected by automatic biochemistry analyzer, The interior diameters of the portal vein, the spleen vein and the thickness of the spleen were also measured with ultrasonography.</p><p><b>RESULTS</b>16 patients (14.16%) whose serum fibrosis markers were inconsistent with histological stage of liver fibrosis were found. Their serum fibrosis markers were not correlated with staging of liver fibrosis (P>0.05), but were positively correlated with inflammation grade (x(2)=12.07, P<0.05), at same time, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase(AST), gamma-glutamyltransferase (GGT) and globulin (GLB) decreased obviously, from 89.28 U/L +/- 64.25 U/L to 49.31 U/L +/- 26.75 U/L (t=2.45, P<0.05), 66.10 U/L +/- 42.30 U/L to 40.83 U/L +/- 22.40 U/L (t=2.33, P<0.05), 86.26 U/L +/- 70.36 U/L to 48.99 U/L +/- 29.96 U/L (t=2.08, P<0.05) and 32.13 g/L +/- 5.18 g/L to 28.05 g/L +/- 3.47 g/L (t=3.03, P<0.01) respectively. And the level of albumin (ALB) and the ratio of albumin and globulin (A/G) increased evidently, from 42.34 g/L +/- 4.81 g/L to 46.19 g/L +/- 3.61 g/L (t=3.06, P<0.01) and 1.35 +/- 0.28 to 1.63 +/- 0.26 (t=3.70, P<0.01). But the serum level of alkaline phosphatase (ALP), total bilirubin (TBil), total protein (TP), the width of main portal vein, the width of splenic vein and the thickness of the spleen did not change clearly (P>0.05).</p><p><b>CONCLUSION</b>As diagnostic markers, serum fibrosis markers as well as inflammation grade and liver function should be taken into account.</p>