Clinical study of serum vitamin D in chronic HBV infected patients / 西安交通大学学报(医学版)

Objective: To assess the level of serum 25(OH)D in patients infected with chronic hepatitis B virus (HBV) at different stages so as to explore their clinical significance. Methods: We quantified the levels of serum 25(OH)D from 246 chronic HBV-infected patients (chronic hepatitis B, n=90; compensated liver cirrhosis, n=70; and decompensated liver cirrhosis, n=86) in the study. Serum 25(OH)D level, liver function, coagulation index, and HBV markers were tested in all the patients. Child-Pugh grade and FIB-4 score of liver fibrosis were calculated in patients with liver cirrhosis. Results: ① The mean serum level of vitamin D in chronic HBV infection patients was significantly lower than that in healthy controls [(14.37±7.18)ng/mL vs. (23.60±6.20)ng/mL, P<0.05]. Serum 25(OH)D level in patients with decompensated cirrhosis was obviously lower than that in patients with chronic hepatitis B [(15.22±6.84)ng/mL, P<0.001] and compensated cirrhosis [(17.16±7.55)ng/mL, P<0.001]. ② Serum 25(OH)D had a positive correlation with ALB level, but a negative correlation with PT level. Serum 25(OH)D level in liver cirrhosis patients was significant negatively correlated with FIB-4 scores. ③ Of the liver cirrhosis patients, 91.9% patients in Child-Pugh C group, 81.6% patients in Child-Pugh B group, and 67.1% patients in Child-Pugh A group had serum 25(OH)D level deficiency. Univariate and multivariate logistic regression analysis showed that CHB patients with FIB-4 score≥1.45 were more likely to develop compensated liver cirrhosis. Compensated liver cirrhosis patients with serum 25(OH) D level <20 ng/mL had the high possibility of decompensation cirrhosis and more severe hepatic injury. Conclusion: Vitamin D deficiency is prevalent in chronic HBV-infected patients, especially in those with decompensated cirrhosis. Vitamin D level can be used as an indicator to assess the severity of chronic HBV infection.

Relationship between the expression of HBV mRNA in embryos and father-to-infant HBV transmission / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate father-to-infant transmission of hepatitis B virus (HBV) by detecting HBV mRNA in the IVF embryos with paternal HBV infection.</p><p><b>METHODS</b>We collected 18 discarded IVF embryos (9 cases) with paternal chronic HBV infection, and detected HBV mRNA in the embryos by single-cell RT-PCR.</p><p><b>RESULTS</b>HBV mRNA positive signals were found in 1 of the 18 embryos with paternal serum HBV positive markers (5.6%), but no specific HBV mRNA signals were observed in the 84 embryos of the negative control group. Follow-up visits revealed no significant difference between the experimental and negative control groups either in the rate of clinical pregnancy (P > 0.05) or in that of early abortion (P > 0.05). The IVF embryo with paternal HBV mRNA positive signals was successfully implanted, but early abortion occurred. HBV infection was not transmitted to progeny in either of the two groups.</p><p><b>CONCLUSION</b>The positive results of HBV mRNA indicate that HBV can get into early-cleavage embryos through sperm and replicate there, which may be the main channel of father-to-infant transmission. HBV may interfere with the development of embryos, and even result in abortion and other adverse outcomes.</p>

Construction of a bait plasmid containing HBV PreS1 gene in a yeast two-hybrid system and evaluation of its toxicity and self-activation / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a yeast expression vector of hepatitis B virus (HBV) PreS1 gene using the Sos-recruitment system (SRS), and evaluate the effect of the expression product on the growth of the yeast cells and activation of the reporter gene.</p><p><b>METHODS</b>The coding sequence of HBV preS1 was amplified by PCR and cloned into the yeast expression plasmid pSos. The recombinant bait plasmid pSos- PreS1 was verified by sequencing before transformation into competent yeast cells. The effects of the expression product on the yeast cell growth and activation of the reporter gene were evaluated.</p><p><b>RESULTS</b>The yeast expression vector of HBV PreS1 gene was constructed successfully. The recombinant bait plasmid showed no toxic effect on yeast cdc25H cells without a self-activation of the reporter gene.</p><p><b>CONCLUSION</b>The SRS can be used to study the proteins interacting with HBV PreS1 protein and provides a means for obtaining insight into the pathogenic mechanism of HBV.</p>

Cellular localization of HCBP1 and its interaction with HCV core protein in vivo / 南方医科大学学报

<p><b>OBJECTIVE</b>To analyze the interaction of hepatitis C virus (HCV) core protein with HCBP1 and observe the expression and cellular localization of HCBP1.</p><p><b>METHODS</b>The cDNA fragments encoding HCV core protein and HCBP1 were amplified by PCR and subsequently cloned into pGEM T vector, respectively. After sequence verification, the two recombined vectors were respectively subcloned into two hybrid plasmids, pM and pVP16. pM-core, pVP16- HCBP1 and the reporter vector pG5CAT were co-transfected into COS-7 cells, and the interaction between HCV core protein and HCBP1 was assayed by detecting CAT gene expression after 48 h. The expression and subcellular localization of the fusion protein in the transfected COS-7 cells were analyzed by Western blotting and fluorescence microscopy, respectively.</p><p><b>RESULTS</b>CAT-ELISA showed that the absorbance of the co-transfection group was significantly higher than that o f the negative control groups but lower than that of the positive control group. Western blotting confirmed the expression of fusion protein in the transfected COS-7 cells. Fluorescence microscopy showed that the fusion protein was distributed mainly in the cytoplasm, and in contrast, diffuse EGFP expression was detected in COS-7 cells transfected with the empty vector.</p><p><b>CONCLUSION</b>Mammalian two-hybrid assay confirms the capacity of HCBP1 to bind HCV core protein, and the expression vector for HCBP1-EGFP fusion gene has been constructed successfully and expressed in COS-7 cells.</p>

A Clinic Research About Bronchoscopic Lung Volume Reduction By ?-cyanoacrylate / 中国实用内科杂志

Objective To research the method and effect about the bronchoscopic lung volume reduction by ?-cyanoac- rylate in the therapy of chronic obstructive pulmonary disease (COPD).Methods The 14 patients had been examined bosoms by CT before the operation and determined the type of emphysema and the distributing of pneumatocele,had blood gas analyzed and pulmonary function checked.The operation was carried through trachea cannula and intravenous anesthe- sia.When the bronchoscope came to the goal bronchia,we infused the meglumine diatrizoate through the biopsy orifice and approved the location of pneumatocele forward。Then,we infused erythromycin and ?-cyanoacrylate in turn through the biopsy orifice by silica del tube.Results The 3 pneumothorax patients had been removed the drainage tube in 3 days af- ter the operation.8 cases had been counterchecked sternite in one week and the pneumatocele was just like before,among which,1 case developed exudation.1 case had shown pleural thickening in the right-up lung counterchecked sternite 9 months later.1 case been checked the pulmonary function,the FEV_1 enhanced from 24.7% pred before operation to 32. 9% pred after operation one week.3 cases felt polypnea improved greatly and 7 cases felt polypnea improved a little.Con- clusion The bronchoscopic lung volume reduction by ?-cyanoacrylate is a safe,effective and economical method in the therapy of COPD.

Screening of HBeAgTP interacting proteins in hepatocytes with yeast-two hybrid technique / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To screen proteins in hepatocytes interacting with HBeAg transactivated protein (HBeAgTP) with yeast-two hybrid technique for investigating the biological functions of HBeAgTP.</p><p><b>METHODS</b>Suppression subtractive hybridization (SSH) and bioinformatics techniques were used for screening and cloning of the target genes transactivated by HBeAg. The HBeAgTP gene was amplified by polymerase chain reaction (PCR) and HBeAgTP bait plasmid was constructed with yeast-two hybrid system 3, and then transformed into yeast AH109. The transformed yeast mated with yeast Y187 containing liver cDNA library plasmid in 2 x YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-gal for selecting two times and screening. After extracting and sequencing of plasmid from blue colonies, the results were analyzed by bioinformatics.</p><p><b>RESULTS</b>HBeAgTP gene was successfully cloned and expressed in yeast cells. Fifteen genes in twenty-four positive colonies were obtained using yeast-two hybrid technique.</p><p><b>CONCLUSION</b>HBeAgTP conjugated protein genes were successfully cloned, along with the genes involved in transcription and translation of proteins, immunoloregulation, materials and energy metabolism in vivo.</p>