OBJECTIVE:To observe the efficacy and safety of methylprednisolone combined with entecavir in the treatment of primary nephrotic syndrome with hepatitis B virus infection. METHODS:23 primary nephrotic syndrome patients with hepatitis B virus infection were given 0.5 mg/times Entecavir tablet,orally,once a day,for 2 weeks,and added 0.8mg/(kg.d)Methylpredniso-lone tablet,orally,once a day,for 8 weeks,then the dose was maintained for 2-3 months every other day,and then decreased 4 mg for 4 weeks every other day,and the dose was decreased 4 mg every 2-4 weeks,until drug withdrawal. Entecavir tablet was used in the whole process of Methylprednisolone tablet,the dose was adjusted and gradually stopped based on HBV-DNA within 3 months of Entecavir tablet withdrawal. Liver and renal efficacy,24 h urine protein excretion,serum albumin(ALB),alanine ami-notransferase(ALT),aspartate aminotransferase(AST)levels before and after treatment of 2,6,12 and 15 months,and the inci-dence of adverse reactions of all patients were observed. RESULTS:The liver and renal efficacy were 100%;after treatment of 2, 6,12 and 15 months,24 h urine protein,ALT and AST were significantly lower than before and gradually decreased by time, ALB was significantly higher than before and gradually increased by time,the differences were statistically significant(P<0.05);the incidence of adverse reactions was 43.48%,and there were no obvious adverse reactions and no renal dysfunction. CONCLU-SIONS:Methylprednisolone combined with entecavir has significant efficacy in the treatment of primary nephrotic syndrome with hepatitis B virus infection,with good safety.

Objective To explore clinical effect of the stellate ganglion block combined buflomedil in the treatment of vertebral artery type of cervical syndrome (CSA).Methods One hundred and twenty cases of CSA were included in the study,and randomly divided into two groups.Study group (60 cases)patients were treated by stellate ganglion block therapy combined with buflomedil intravenous;the controlled group (60 cases)was treated with buflomedil intravenous therapy only.In the treatment,vertebral-basi-lar artery mean flow velocity (Vm)was measured before and after treatment and comparison of Vm difference was the clinical ba-sis.According to the CSA standard of clinical cure,the clinical curative effect was observed.Results After treatment,the total ef-fective rate of study group was 95.00%,total effective rate of control group was 71.67%,the difference statistically significant (χ2 =24.474,P <0.05).vertebral artery blood flow velocity of the two groups after treatment increased more obvious than that of before treatment,the difference was statistically significant (P <0.05),vertebral artery blood flow velocity after treatment of study group (38.44±2.20)cm/s was significantly higher than that of the control group (34.36±3.50)cm/s,the difference was statisti-cally significant (t=7.645,P <0.05).basilar artery blood flow velocity of the two groups after treatment increased more obvious than that of before treatment,the difference was statistically significant (P <0.05),basilar artery blood flow velocity after treat-ment of study group(56.34±4.10)cm/s was significantly higher than that of the control group (47.69±3.90)cm/s,the differ-ence was statistically significant (t= 11.841,P <0.05).Conclusion The clinical efficacy of stellate ganglion block combined bu-flomedil in treatment of vertebral artery type of cervical syndrome is obvious.The cure rate with respect to the drug treatment has significant advantages and the therapy is worthy of further promotion.

OBJECTIVETo screen and identify dengue virus type 2 specific antigens and establish an enzyme-linked immunosorbent assay (ELISA) for detecting dengue virus type 2 antibody.

METHODSUsing the bioinformatic software DNAstar and ANTHEPROT, we analyzed the hydrophilicity, flexibility, surface probability and antigenicity of dengue virus type 1-4, Japanese encephalitis virus, and Yellow fever virus M and E protein amino acid sequences, and also evaluated the influence of secondary structure. The specific epitopes of dengue virus type 2 were predicted according to the epitope location and amino acid sequence similarity, and the epitope conservation was assessed using the sequence information of different dengue virus type 2 strains in GenBank. Based on the results of bioinformatic analysis, 5 specific epitopes were amplified and inserted into the prokaryotic expression vector pET32a, which were transferred into E. coli Rosetta (DE3) for expression of the proteins. SDS-PAGE and Western blotting were used to identify the expressed proteins and test their antigenicities. The antigen selected by Western blotting was used to establish the ELISA system for dengue virus type 2 antibody detection.

RESULTSBioinformatic analysis predicted 8 possible dengue virus type 2 specific epitopes, and 6 of them were efficiently expressed in E. coli. Western blotting confirmed 1 dengue virus type 2 specific antigen, the ELISA system for dengue virus antibody detection was successfully established using this specific antigen.

CONCLUSIONWe have obtained a dengue virus type 2 specific antigen and established an ELISA system for detection of dengue virus type 2 antibody.

Antibodies, Viral ; immunology ; Antigens, Viral ; immunology ; Computational Biology ; Dengue Virus ; classification ; immunology ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immunodominant Epitopes ; Software