ABSTRACT
Objective@#To investigate the antiviral effect of hepatitis B virus (HBV) S gene-specific anti-gene locked nucleic acid (LNA) in transgenic mice.@*Methods@#A total of 30 HBV transgenic mice were randomly divided into blank control group (5% glucose + liposome), unrelated sequence control group, lamivudine control group, antisense LNA control group, and anti-gene LNA group, with 6 mice in each group. The mice in the lamivudine group were given lamivudine by gavage, and LNA was injected via the caudal vein. Quantitative real-time PCR was used to measure serum HBV DNA, ELISA was used to measure serum HBsAg, RT-PCR was used to measure HBV S mRNA level in the liver, and immunohistochemistry was used to measure the level of HBsAg in hepatocytes.@*Results@#At 3, 5, and 7 days after treatment, there were significant changes in the inhibition rates of HBV DNA (37.18%, 50.27%, and 61.46%, respectively) and HBsAg (30.17%, 44.00%, and 57.76%, respectively) achieved by anti-gene LNA (P < 0.01), and there were significant differences between the anti-gene LNA group and the other four control groups (P < 0.05). In the anti-gene LNA group, the relative mRNA expression of HBV S gene was 0.33 and the percentage of HBsAg-positive hepatocytes was 31%, which were significantly different from these two indices in the control groups (P < 0.05). There were no abnormal changes in liver/renal biochemical parameters and HE staining results.@*Conclusion@#Anti-gene LNA targeting at HBV S gene has a strong antiviral effect in transgenic mice, which provides theoretical and experimental bases for gene therapy for HBV.
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Objective To analyze the MP infection status in children with respiratory tract disease and the correlation with gen‐der ,age ,season and clinical conditions .Methods To investigate the clinical data retrospectively of 655 children with respiratory tract infection from January to December 2013 .Results The positive rate of MP antibody was 48 .09% with a higher incidence in girls than boys ,and those above 3 were more susceptible to it .Winter and spring were the peak seasons .The older group had a higher positive rate of MP antibody ,together with high morbidity of MPP and LP ,the differences were statistically significant (P<0 .05) ,while the younger group was inclined to get higher WBC count and percentage of increasing CRP and neutrophils ,the differ‐ences were statistically significant(P<0 .05) .Conclusion MP is a common pathogenic bacteria causes respiratory tract infection in children above 3 years ,especially in winter and spring .The antibody positive rate rise with age and the infected children are more likely to have pneumonia meanwhile the younger group has higher WBC count ,in which more cases get higher level of CRP and neutrophils .
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Objective To investigate the expression and biological function of miRNA-449 a in lung cancer . Methods A case-control study was conducted in 58 patients diagnosed with lung cancer ( carcinoma and adeno-carcinoma) and normal tissue closely adjacent to tumor.MiRNA-449a simulation was designed and synthesized, was dissolved into two different concentrations as 10 and 20 mg/mL.The expression of miRNA-449a in lung cancer tissues and matched normal tissues were detected by Real time PCR .The expression of luciferase gene was detected by chemiluminescence technique.MiRNA-449a mimics on cell apoptosis was evaluated by MTT assay . Results The mean tissues expression levels of miRNA-449 a in squamous carcinoma group and adenocarcinoma group were 1.48 ±1.63 and 1.52 ±1.54 respectively, and were significantly lower than in control group (2.74 ± 1.55 ) ( P<0.01 ) .The average intensity of fluorescent protein in 10 mg/mL group and 20 mg/mL group were 2 115 ±168 and 1 352 ±159 respectively , and were significantly lower than that in control group ( 4 975 ±115 ) ( P<0.01 ) .Conclusions MiRNA-449 a was down-regulated expression in lung cancer and induced apoptosis .
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This paper is aimed to investigate the inhibitory effects of hepatitis B virus (HBV) preC and C genes-specific antisense locked nucleic acid (LNA) on HBV replication and expression in transgenic mice. The antisense LNA, which was complementary to the preC and C gene region of HBV, was designed, synthesized, and injected into transgenic mice via the tail vein. Serum HBV DNA was tested with real-time PCR, and Serum HBsAg was tested with time-resolved fluorescence immune assay (TRFIA). Then the expression of HBcAg in the liver was detected with immuneohistochemistry. Serum ALB, ALT, BUN and CRea were measured with an antomatic biochemicall analyzer. It was found that 5 days after LNA injection, serum HBV DNA levels in the dual-target group were reduced by 53.72%, and serum HBsAg levels were decreased by 71.57%. These values were significantly higher than those in the control groups (P<0.05) and the expression levels of HBcAg in the liver were significantly lower than those in the control groups (P<0.05). The result also showed that there were no significant differences discovered in serum ALB, ALT, BUN and CR between the experiment groups and the control groups. The present study provides that antisense LNA targeting to both preC and C genes has shown strong inhibition on HBV replication and expression in transgenic mice, and stronger than target at single gene site.
Subject(s)
Animals , Female , Male , Mice , Antiviral Agents , Pharmacology , DNA, Viral , Blood , Gene Targeting , Hepatitis B Core Antigens , Metabolism , Hepatitis B Surface Antigens , Blood , Hepatitis B virus , Genetics , Physiology , Liposomes , Mice, Transgenic , Oligonucleotides , Pharmacology , Oligonucleotides, Antisense , Pharmacology , Virus ReplicationABSTRACT
Objective To investigate the inhibitory effects of hepatitis B virus (HBV) S gene-specific antisense locked nucleic acid (LNA) on HBV replication and expression in HepG_22.2.15 cells,and to screen the effective short sequence of LNA.Methods Four different lengths of short sequence of antisense locked nucleic acid which were complementary to the initiator of HBV S gene were designed,synthesized and transfected by cationic liposomes into HepG_22.2.15 cells.The HBsAg and HBV DNA of supematant was tested by enzyme linked immunoadsorbent assay(ELISA) and real-time fluorescent quantitative PCR(FQ-PCR) at 24,48 and 72 hours after treatment.LNA's cyto-toxicity on cell was evaluated by MTT method.Results Four different lengths of short sequence of LNA inhibi-ted the expression of HBsAg and the replication of HBV DNA with the inhibition rates of 46.58%,54.38%,72.43% ,69.92% and 27.09% ,28.77% ,34.71% ,32.68% respectively after 72 hours.There's no obvious tox-icity on cell.Conclusion Antisense LNA that targeting at HBV S gene has strong inhibition on HBV in vitro,and the optimal length of LNA sequence might be in the range of 15 base to 25 base.It has a therapeutic potential in the treatment of patients infected with HBV.