Detection of double mutant X antigen/antibody in sera of patients with hepatitis B virus infection and its implication / 中华传染病杂志

Objective To explore the correlation between serum hepatitis B virus (HBV) X antigen/antibody (HBxAg-wild/HBxAb-wild,and HBxAg-mutant/HBxAb-mutant) and the disease progression in patients with chronic HBV infection.Methods A direct enzyme immunosorbent asssay (ELISA) was performed to detect HBxAb using recombinant antigen,and a double antibody sandwich ELISA assay to detect HBxAg using monoclonal antibody and specific rabbit polyclonal antibody.HBxAg-wild/HBxAb-wild and HBxAg-mutant/HBxAb-mutant were tested in sera from cases at different stages of chronic HBV infection.A chi-square test was employed to examine statistical significance.Results The positive rates of HBxAg-wild and HBxAg-mutant in the chronic asymptomatic HBV carriers,chronic hepatitis,hepatitis B-related cirrhosis and liver cancer were 6.2％ (2/32),10.7％ (3/28),28.6％ (6/21),43.6％ (17/39) and 3.1％ (1/32),10.7％ (3/28),33.3％ (7/21),48.7％ (19/39),respectively.The positive rates of HBxAb-wild and HBxAb-mutant in the above mentioned groups were 6.2％ (2/32),21.4％ (6/28),38.1％ (8/21),53.8％ (21/39)and 6.2％ (2/32),25.0％ (7/28),42.9％ (9/21),61.5％ (24/39) respectively.The positive rates of HBxAg-wild and HBxAg-mutant were not significantly different among the above groups (x2 =0.871,0.780,0.565 and 0.317,respectively; all P＞0.05) ; The positive rates of HBxAb-wild and HBxAb-mutant were also similar among all the groups (x2 =0.780,0.709,0.580 and 0.210,respectively; all P＞0.05).The positive rates of HBxAg-wild,HBxAb-wild,HBxAg-mutant,HBxAb-mutant in patients with low viral loads (HBV DNA＜1 × 104 copy/mL) were 36.5％ (23/63),44.4％ (28/63),42.9％ (27/63) and 54.0％ (34/63),respectively,those in patients with high viral loads (HBVDNA≥1×104 copy/mL) were 8.8％ (5/57),15.8％ (9/57),5.3％ (3/57) and 14.0％ (8/57),respectively.The positive rates of HBxAg and HBxAb were significantly higher in cases with low viral loads than those with high viral loads (x2 =12.869,11.522,22.556 and 20.976,respectively; all P＜0.05).The positive rates of HBxAg-wild,HBxAb-wild,HBxAg-mutant,HBxAb-mutant in the HBeAg positive group were 21.7％ (18/83),30.1％ (25/83),22.9％ (19/83) and 32.5％ (27/83),respectively,while those in the HBeAg negative group were 27.0％ (10/37),32.4％ (12/37),29.7％ (11/37) and 40.5％ (15/37),respectively.No significant difference of HBxAg/HBxAb positive rates between HBeAg positive group and HBeAg negative group was noticed (x2 =0.408,0.064,0.638 and 0.722,respectively; all P＞0.05).Conclusions The antigenicity and specificity of HBV X protein remains similar after the occurrence of A1762T/G1764A double mutant in X gene.It is also found that the positive rates of HBxAg and HBxAb increase with disease progression.HBxAg/HBxAb might be promoting factors for tumorigenesis in chronic HBV infection.HBxAg and HBxAb might have negative influence on HBV replication.

Correlation of hepatitis B virus X antigen expression with liver cell apoptosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the roles of hepatitis B virus (HBV) X antigen (HBxAg) in development of HBV-related liver diseases and carcinogenesis.</p><p><b>METHODS</b>Liver tissues were collected from patients with HBV infection (HBV carriers, n = 14; chronic hepatitis B (CHB), n = 24), HBV-related liver cirrhosis (LC, n = 20), or hepatocellular carcinoma (HCC, n = 20). Immunohistochemistry was used to detect the expression of HBxAg and the host apoptosis-related genes Fas and Fas ligand (Fas-L). The correlations of HBxAg with HBV DNA level in serum, inflammation grade, and fibrosis stage were statistically analyzed. Liver inflammation grade and fibrosis stage were in accordance with Knodell standard. x2test and Fisher's exact test were adopted in count data, x2split method was adopted in pariwise comparisons between multiple samples, Rank-sum test was adopted in ranked data, Spearman rank correlation analysis was adopted in correlation analysis.</p><p><b>RESULTS</b>The rates of HBxAg-positivity were similar between the patients with HBV infection (71.1%), LC (60.0%), and HCC (65.0%) (x2= 0.754, P = 0.686). The rates of Fas- and Fas-L-positivity in liver cells were also similar between the three groups (Fas: 28.9% vs. 20.0% vs. 5.0%, x2= 4.667, P = 0.101; Fas-L: 36.8% vs. 50.0% vs. 60.0%, x2= 2.988, P = 0.225). However, the positive rate of Fas in lymphocytes of liver tissue was significantly higher in the HCC patients than in the HBV-infected patients (90.0% vs. 68.4%, Z = -4.360, P = 0.00001). The expressions of HBxAg and Fas-L corresponded to regions of severe inflammation in tissues from LC patients and some HCC patients. Furthermore, the expression of HBxAg was positively correlated with Fas (r = 0.304, P = 0.02) and Fas-L (r = 0.368, P = 0.004) in the HBV-infected patients and LC patients, and the expression of Fas was positively correlated with that of Fas-L (r = 0.448, P = 0.0004). Patients with high and medium loads of HBV DNA showed significantly higher rates of HBxAg-positivity than those with low loads (88.9% and 69.2% vs. 26.7%, P less than 0.05).</p><p><b>CONCLUSION</b>In the early stage of chronic HBV infection, HBxAg may induce liver cell apoptosis by up-regulating Fas expression, and in the later stage, HBxAg may induce immune escape by up-regulating Fas-L expression in liver cells. Together, HBxAg and high HBV DNA load may promote chronic HBV infection and progression to hepatocarcinogenesis.</p>

Synthesis and antitumor activity of novel 2-(1-substituted-piperidin-4-ylamino)quinazolines as antitumor agents / 药学学报

A variety of novel 2-(1-substituted-piperidine-4-ylamino)quinazoline derivatives were prepared and their antiproliferative activities on five cancer cell lines were evaluated by MTT assay. Quinazolines 4j-4l, 5a, 5b and 5d bearing a small hydrophobic alkyl group on piperidine ring exhibited potent antitumor activities with IC50 values at micromolar level. Compound 41 displayed significant in vivo antitumor activity with 72.9% inhibition on H22 tumor growth and 80% inhibition on Lewis lung cancer growth at a dose of 200 mg x kg(-1).

The cloning, expression, purification and immunological identification of wild-type and mutant hepatitis B virus X gene in pGEX-6P-2 system / 病毒学报

To settle the foundation for the future research on the influence of wild and mutant (A1762T/ G1764A) HBV X gene on the progress of chronic HBV infection and hepatic tumorigenicity, wild and mutant (A1762T/G1764A) HBxAgs expression system was constructed. The wild and mutant (A1762T/ G1764A) HBV X genes were amplified with polymerase chain reaction (PCR) from HBV genome were inserted into pGEX-6P-2 and confirmed by sequencing respectively. Prokaryotic expression vectors pGEX-6P-2-hbvx(w) and pGEX-6P-2-hbvx(m) (A1762T/G1764A) were constructed and transformed to Trans1-blue; wild and mutant HBxAgs were expressed through IPTG induction respectively; after refolding of inclusion body, the wild and mutant HBxAgs were purified with GSTrap FF; and analysised by SDS-PAGE, Western blot and ELISA. SDS-PAGE analysis showed that the expression system was able to express target protein efficiently; the concentrations of purified wild HBxAg and mutant HBxAg were 4.88 mg/mL and 5.07 mg/mL respectively; Western blot analysis certified both the wild HBxAg and the mutant HBxAg could be recognized by the same monoclonal antibody against HBxAg; the two expressed fusion antigens coated in microtiter plate were able to react with the sera of HBV infected patients but not with the sera from healthy donors in ELISA. Results demonstrated that we successfully established a system for expression of hepatitis B x antigen and lay the foundation for further research on the role and molecular mechanisms of the mutant HBxAg in the progress of chronic HBV infection and hepatic tumorigenicity.

Expression and clinical significance of matrix metalloproteinase in gastric stromal tumor / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the expression and clinical significance of matrix metalloproteinase (MMP) in gastric stromal tumor (GST).</p><p><b>METHODS</b>MMP-2 and MMP-9 expression were determined by immunohistochemistry in tumor tissues in 44 patients with GST, and their relationship with clinicopathologic factors of the neoplasm was also investigated.</p><p><b>RESULTS</b>MMP-2 and MMP-9 were expressed in the cytoplasm in 84.1% (37/44) and 81.8% (36/44) of tumors, respectively. The positive rates of MMP-2 and MMP-9 increased significantly in parallel to the increase in tumor malignancy (P < 0.05) and associated with pattern of tumor growth, tumor size, and centre necrosis (P < 0.05). In addition, there was a statistically significant positive correlation between the expression of the two markers in GST (r = 0.6523, P < 0.05). Furthermore, the 5-year postoperative survival rates of patients with positive expressions of MMP-2 and MMP-9 were significantly lower than those of patients with negative expressions of the two markers (P < 0.05).</p><p><b>CONCLUSION</b>Over expression of MMP-2 and MMP-9 can be served as objective markers to judge the malignant degree and, to predict the prognosis of patients with GST.</p>

The effect of postoperative chemotherapy after esophagectomy on the survival rate to patients with original squamous cell carcinoma of esophagus / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the prognostic factors affecting the survival rate after extended radical esophagectomy with three-field lymph node dissection for squamous cell carcinoma and the effect of postoperative chemotherapy.</p><p><b>METHODS</b>Out of 126 patients with original squamous cell carcinoma of esophagus who accepted extended radical esophagectomy with three-field lymph node dissection from 1987 - 1992 in a hospital, 97 of them were included in this study. Data on the clinical/pathological characters and post surgery survival records of the subjects' were collected. Survival analysis methods included Kaplan-Meier, Log-rank test and Cox multivariable model and the effects of postoperative chemotherapy were analyzed for patients in early and late stages.</p><p><b>RESULTS</b>There was no significant difference in clinical and pathological character between those patients only undergone surgery and patients accepting postoperative chemotherapy. The size of tumor, grade of differentiation of the tumor cells, infiltration deepness, with or without lymph node metastasis, expression of nm23 and EGFR and treatment after surgery etc. were correlated with the survival rate. For patients in early tumor stage, postoperative chemotherapy with cisplatin and 5-FU after surgery seemed to be a risk factor. For patients in late stage, postoperative chemotherapy with cisplatin and 5-FU after surgery did not seem to improve survival rate.</p><p><b>CONCLUSION</b>It is imperative to study on the effect of adjuvant postoperative chemotherapy to patients, especially those at early stage with squamous cell carcinoma of esophagus. Doctors must be scrupulous when making decisions.</p>

Expression of vascular endothelial growth factor in synovium of rats with adjuvant arthritis / 实用儿科临床杂志

Objective To investigate the expression of vascular endothelial growth factor(VEGF)in synovium of rats with adjuvant arthritis and the relationship between the histopathologic score and the expression of VEGF.Methods Adjuvant arthritis was established in Wistar rats by inoculating complete Freund's adjuvant(CFA). We calculated the arthropathologic score and the expression of VEGF mRNA and protein at different stages after CFA inoculation.Results In model group the arthropathologic score and expression of VEGF protein in synovium increased significantly all the time (P