Objective To observe the suppression of HBsAg and HBeAg expression by DNAzyme and LNAzyme located at HBV pre-area of HBV.Methods Eecoding sequence of(10-23 DNAzyme) thiolmodificated 10-23DNAzyme and LNAzyme that were directed against Pre C/C region of HBV were designed and synthesized.Experimental groups and control groups were set up.The experimental groups included 10-23 DNAzyme group,(S-10-23) DNAzyme group and LNAzyme group.The control groups include blank control group,simple lipofectamine group,simple 10-23DNAzyme group and random 10-23 DNAzyme group.In the dosege of 0.16,0.64,1.28,1.60,(1.92 ?mol?L~(-1)) and the time of 12,24,36,48,60,72,84 and 96 h,the suppression of HBsAg and HBeAg expression by 10-23 DNAzyme and LNAzyme in 2.2.15 cells were studied.Results The suppression of HBsAg and HBeAg expression by 10-23 DNAzyme and LNAzyme in 2.2.15 cells were significant.The inhibitory effects caused by LNAzyme was more significant than that by thiolmodified 10-23 DNAzyme whose inhibitory effects were more significant than that of 10-23 DNAzyme.The inhibitory rates of LNAzyme and 10-23 DNAzyme thiolmodification reached(91.6?8.4)%,(78.4?2.0)% on HBsAg,respectivelly and(90.1?5.2)%,(76.4?4.8)% on HBeAg.The inhibitory effects of LNAzyme and thiolmodification of 10-23 DNAzyme were found 12 h after they were added to 2.2.15 cells,and optimized at 48 h,effective inhibitory time for LNAzyme was 84 h,for thiolmodification 10-23 DNAzyme was 72 h.Addition of LNAzyme and 10-23 DNAzyme to 2.2.15 cells didn′t exert cytotoxicity.Conclusion 10-23 DNAzyme and LNAzyme have demonstrated significant inhibitory effects on the HBsAg and HBeAg expressions in 2.2.15 cells.Morever,the inhibitory effects of LNAzyme is more significant than that of DNAzyme.LNAzyme is a specific anti-HBV therapeutic agent.

Cell apoptosis is an important biological process. It plays an essential role on cell growth and response for outside stimulations. This review focused on the recent advancement of apoptotic signal transduction, enzymological mechanisms, the function of mitochondria in cell apoptosis as well as the regulation of genes.

Objective To observe the suppression of special shRNA producing plasmid to hepatitis B virus(HBV) S gene and C gene on HBV replication and expression in HepG2.2.15 cells.Methods pSilenceCircle-U6 including pol Ⅲ promoter was used to construct HBV special shRNA producing plasmid as SC-S and SC-C.The experimental groups included SC-S group,SC-C group,unrelated control SC-N group and blank control group.With different dosages and at different time,shRNA producing plasmid was transfected into HepG2.2.15 cells.HBeAg and HBsAg in the culture media was detected by ELISA assay and HBV DNA in the culture media was measured by dot blotting assay. Results The recombinant shRNA producing plasmid with target sequence was constructed successfully.The inhibitory rates of HBeAg and HBsAg expressions by SC-S were much higher than those by SC-C.The inhibitory effects of HBeAg and HBsAg expressions were increased when the dose of SC-S was greater.The inhibitory effects of HBeAg and HBsAg expressions by SC-S were significant on the 3rd day after transfection and the inhibitory effect was the strongest on the 6th day.The inhibitory rate was still higher on the 9th day after transfection.Dot blotting assay showed the inhibitory effect of HBV replication by SC-S was greater than that by SC-C.Conclusion The shRNA producing plasmid with HBV S gene and C gene can be highly effective to inhibit the replication and expression of HBV.

Objective To study the effect of specific hammerhead ribozyme (Rz) to hepatitis C virus (HCV) in vitro. Methods Rz 1 Rz 2 were designed to cleavage at 5′-NCR nucleotide positions under 136～160 and 313～337, Rz 3 was designed to cleavage at C region nucleotide position under 373～388. As a control, cleavage deficient Rzm that have A→G point mutations in the catalytic loop of the hammerhead domain. 32P-labeled transcript of target HCV RNA was incubated with gel-purified Rz ( Rz 1, Rz 2, Rz 3 and Rzm ) respectively at different concentration based on specified condition and autoradiographed after denaturing gel-electrophoresis. Results Except Rzm, Rz 1 Rz 2 Rz 3 were active at 37 ℃ and more so at higher concentration, and more so with cleavage site nearly to the HCV initial code. Conclusions The HCV specific hammerhead ribozyme can be designed in vitro, further study about cleavage in vitro and in vivo will continue.

Objective To assess the effects of medicated permeation with traditional Chinese medicine Shujin-Zhitong liquid combined with ion tophoresis on osteoarthritis(OA) of rabbit knee joint in vivo.Methods The modified Hulth OA model was prepared with rabbits by anterior cruciate ligament desmotomy plus medial meniscus resection.Twenty-four model rabbits were randomly divided into three groups: Chinese medicine group: treated with Chinese medicine Shujin-Zhitong liquid by ion tophoresis for 20 minutes per day;positive control group: glucosamine hydrochloride was administered orally per day;blank control group: no treatment was administered to the model rabbits.Serum nitrogen monoxidum(NO) concentration was determined at the time points of prior model making,prior treatment,and 4 and 8 weeks of treatment.At 4 and 8 weeks of treatment,the apoptotic rate of chondrocytes was determined by flow cytometry,and the gene expressions of collagen Ⅱand aggrecan were determined by reverse transcription polymerase chain reaction(RT-PCR).Results The contents of serum NO in Chinese medicine group and positive control group were lower than that in blank control group at 4 weeks after treatment,and the content of serum NO in Chinese medicine group was lower than those in positive control group and blank control group at 8 weeks after treatment.The apoptotic rates in Chinese medicine group and positive control group were lower than that in blank control group at 4 and 8 weeks after treatment.Gene expression levels of collagen Ⅱ and aggrecan in Chinese medicine group and positive control group were higher than that in blank control group at 4 weeks after treatment,while the gene expression level in Chinese medicine group was higher than those in positive and blank control groups at 8 weeks after treatment.Conclusions The treatment of medicated permeation with traditional Chinese medicine Shujin-Zhitong liquid combined with ion tophoresis has shown good therapeutic effect on osteoarthritis(OA) of rabbit knee joint,the mechanism might be the inhibition of chondrocytes apoptosis and reduced secretion of inflammatory factors,and the improvement of cartilage repair by up-regulating related gene expression of chondrocyte.

Objective To detect the transfusion transmitted virus-like mini virus(TLMV) sequence gene in patients with chronic hepatitis and to study the TLMV infection in population.Methods Two sets of primers in the most conservative regions of Japan strains TLMV-CBD279 and CBD231 were designed to amplify TLMV templete extracted from sera of patients with chronic hepatitis B using PCR.PCR products were cloned into pGEMR-T vector and sequenced.Results The results showed that 72%of TLMV sequence isolated in China was identical to that in Japan,suggesting that there was TLMV infection in patients with chronic hepatitis B in China.The patients with hepatitis virus C and the healthy blood donor had the highest infectious rates of TLMV.Conclusion TLMV infection exists in patients with chronic hepatitis B in China.

Objective:To analyze the curative effects of laparoscopic radical prostatectomy for the prostate cancer and the its effect on the level serum hormones levels.Methods:86 patients with prostate cancer were selected and divided into the control group and the observation group with 43 cases in each group according to the draw method.The control group was treated by open radical prostatectomy,while the observation group was treated by laparoscopic radical prostatectomy.The operation indicators,serum follicle stimulating hormone (FSH),luteinizing hormone (LH),dihydrotestosterone (DHT),free testosterone (FT),total testosterone (T),prostate specific antigen (T-PSA,F-PSA) levels,CD3+,CD4+,CD8+,CD4+/CD8+ before and after the surgery as well as the occurrence of postoperative complications were compared between two groups.Results:The operation time of observation group was significantly longer than that of the control group,and the blood loss,bowel function recovery time,hospital stay,pain scores of observation group was significantly lower or shorter than those of the control group (P＜0.05).No significant difference was found in the serum FSH,LH,DHT,FT,T,T-PSA,F-PSA levels before and after the surgery between two group(P＞0.05).The CD3+,CD4+,CD4+/CD8+ of observation group was significantly higher than those of the control group (P＜0.05),the CD8+ of observation group was lower than that of the control group(P＜ 0.05).The incidence rate of postoperative complication in the observation group was significantly lower than that of the control group (P＜0.05).Conclusion:Laparoscopic radical prostatectomy could play a similar effect with the open surgery on the tumor control,it could improve the level of serum testosterone,immune function with high safety.

Acetaldehyde ; metabolism ; Alcoholics ; Alcoholism ; blood ; immunology ; Antibodies, Anti-Idiotypic ; Humans ; Immunoglobulin A ; immunology ; Immunoglobulin G ; immunology ; Immunoglobulin M ; immunology ; Protein Processing, Post-Translational

Hepatitis C ; genetics ; Humans ; RNA, Catalytic ; genetics ; RNA, Small Nuclear ; genetics

Objective To evaluate the effect of coagulation function changes on the incidence of acute kidney injury (AKI) after liver transplantation. Methods Clinical data of 245 liver transplant recipients who met the inclusion and exclusion criteria were retrospectively analyzed. According to the incidence of AKI after liver transplantation, all recipients were divided into the AKI group (n=99) and non-AKI group (n=146). The incidence of AKI after liver transplantation was summarized. Perioperative parameters of the recipients were collected. The risk factors of AKI after liver transplantation were assessed by univariate and multivariate analysis. Results Among 245 recipients undergoing liver transplantation, 99 cases developed AKI after operation with an incidence rate of 40.4%. Preoperative serum creatinine levels of the recipients and the in-hospital fatality were relatively high in the AKI group (all P < 0.05). Compared with the recipients in the non-AKI group, those in the AKI group presented with significantly higher liver function parameters within postoperative 24 h, significantly decreased levels of stage Ⅱ coagulation parameters including coagulation factorsⅤ, Ⅶ, Ⅸ, Ⅹ, Ⅻ and protein S, protein C and antithrombin Ⅲ, evidently elevated prothrombin time international normalized ratio (PT-INR), remarkably increased stage Ⅲ coagulation parameters including D-dimer and fibrin degradation product (FDP) levels and considerably decreased fibrinogen (FIB) level (all P < 0.05). Thrombelastogram showed that the R value was increased, the α angle was decreased and the coagulation time was prolonged in the AKI group (all P < 0.05). Logistic regression analysis demonstrated that the increased R value of postoperative thrombelastogram [odd ratio (OR) 1.116, 95% confidence interval (CI) 1.018-1.223, P=0.019], and decreased levels of antithrombin Ⅲ (OR 0.974, 95%CI 0.955-0.993, P=0.007) were the independent risk factors of incidence of AKI after liver transplantation. Conclusions The incidence of AKI after liver transplantation is high, which is associated with the coagulation function changes of the recipients. Decreased coagulation factor activity (increased R value) and declined antithrombin Ⅲ level are the independent risk factors of AKI in liver transplantat recipients.