Objective To construct and identify the specific interference vector for second mitochondria -derived activator of caspase (SMAC)gene in mice and perform lentiviral packaging.Methods According to the SMAC gene sequences in mice,three small interfering RNAs (siRNAs)(siRNA1,siRNA2,and siRNA3)and one negative control sequence (siRNAn)were designed and synthesized,and mouse hepatocytes were transfected with the above siRNAs using Lipofectamine 2000.The inhibitory effects of these siRNAs on SMAC mR-NA expression were evaluated by real-time PCR,and the optimal siRNA was screened out accordingly.Oligo DNA was synthesized based on the optimal siRNA and then connected to GV1 15 vector to obtain recombinant interference plasmid.The candidate clones were identified by PCR and sequenced.The recombinant interference plasmid,as well as pHelper 1.0 and pHelper 2.0,was used to transfect 293T cells for lentivirus packaging.Then,cell supernatants were collected and concentrated,and the lentiviral titer was determined by gradient dilution. Comparison between groups was made by analysis of variance,and multiple comparisons were made by SNK-q test.Results The three siRNAs had different inhibitory effects on SMAC mRNA expression in hepatocytes;siRNA1 showed the strongest inhibitory effect,with an inhibition efficiency of 70.3%.Based on siRNA1 ,the oligo DNA was successfully synthesized and correctly connected to the lentiviral vec-tor,as proved by sequencing.The lentiviral titer was 6 ×108 TU/ml,as determined by gradient dilution.Conclusion The recombinant lentivirus that inhibits SMAC expression in mice has been constructed successfully,laying the foundation for further studies on the role of SMAC gene in liver failure.

Objective To determine sexual function in 612 male addicts with methadone maintenance therapy (MMT),analyze its cause and explore the treatment strategies.Methods Self-made questionnaire and International Index of Erectile Function-5 ( IIEF-5 ) were used in 612male addicts receiving MMT.ResultsThe number of addicts who felt worse sexual function was significantly more than those who felt better sexual function (P ＜0.01 ).About 88.6％ addicts were dissatisfied with their current sexual function,90.7％ of whom were willing to continuously receive MMT.The number of addicts suffering from hyposexuality and erectile dysfunction (ED) was significantly increased at post-MMT than at pre-MMT ( P ＜ 0.01 ),while the number of addicts with normal sexuality and inhibitive ejaculation decreased significantly (P ＜0.01 ).Both the dose of methadone and the age of subjects were negatively correlated with IIEF-5 scores.Correlation between the duration of MMT and IIEF-5 scores was not found.ConclusionMMT might deteriorate sexual function in male addicts.To improve the compliance of MMT and sexual function in male addicts,the dose of methadone should be adjusted to minimally effective one.

ObjectiveTo explore the optimal acupuncture-moxibustion treatment protocol in treating diabetic gastroparesis. MethodA hundred patients were randomized into a body-acupuncture group, an electroacupuncture group, a hydroacupuncture group, and an acupuncture-moxibustion group, allto receive 3 treatment courses. The clinical symptoms and sign scores were observed before and after the intervention in the four groups.ResultThe markedly-effective rate of the acupuncture-moxibustion group was significantly higher than that of the other three groups (P<0.05). Regarding the symptom score for bloating, poor appetite, belching, upper abdominal pain, and nausea, the improvements in the acupuncture-moxibustion group were more significant than that in the other three groups (P<0.05,P<0.01).ConclusionCombination of acupuncture and moxibustion is the optimal treatment protocol for diabetic gastroparesis.