Objective To analyze the outcome of patients receiving intrauterine insemination with husband sperm,in order to evaluate the effect of relative factors on pregnancy rate after intrauterine insemination. Methods Ninety-eight infertile couples who received intrauterine insemination in the Affiliated Hospital of Nantong University from March 2013 to May 2014 were selected as our subjects and 181 cycles were included. The information including clinical factors including maternal age,infertile time,infertile causes, ovulation induction protocol,time of insemination and postwash total motitle sperm(TMS)and pregnancy rate were recorded. Results (1)Totally 26 patients received clinical pregnancies,and clinical pregnancy rate(CPR) was 14. 36% per cycle. With age increase pregnancy rate decreased( χ2 = 1. 654 9,P = 0. 647).(2)The pregnancy rate of the patients was the same within the infertile time( χ2 = 1. 588 5,P = 0. 662).(3)The pregnancy rate of the patients with secondary infertility was lower than that of the patients with primary infertility,but there was no significant difference(χ2 = 0. 923 3,P = 0. 337).(4)The pregnancy rate of ovulation induction cycles was lower than that of nature cycles,but there was no significant difference(χ2 = 2. 222 0,P= 0. 136).(5)Postwash TMS was showed the same trend(χ2 = 0. 643 4,P = 0. 422). Conclusion In terms of intrauterine insemination with husband sperm,age,infertile time,infertile types,ovulation induction protocol and posrwash TMS can affect pregnancy rate,and the effects of various factors should be considered comprehensively in the process of therapy.

Objective To analyze the variation of intestinal microflora in patients with colorectal cancer by SYBR GreenⅠreal-time fluorescence quantitative PCR and reveal the role and significance of intestinal microflora in the colorectal cancer-associated molecular pathogenesis.Methods A set of 16S rRNA gene group of species-specific primers for Bifidobacterium spp.,Lactobacillus group,Escherichia coli,and ddl gene-targeted species-specific primers for Enterococcus faecalis and feces Enterococcus were designed.Patients with colorectal cancer(colorectal cancer group,n=30) and healthy volunteers(normal control group,n=30) were included and whose feces were collected to extract bacterial genome DNA.SYBR GreenⅠ real-time fluorescence quantitative PCR was used to analyze the five mentioned bacterial amounts.Results Level of Bifidobacterium spp.(4.52?0.49) and Lactobacillus group(5.46?0.12) in colorectal cancer group were significantly lower than those(9.25?0.83 and 7.45?0.37) of normal control group(P

Objective To discuss human papilloma virus(HPV)prevalence and HPV genotypes distribution and the infection factors,to provide scientific guidance for the prevention and treatment of cervical cancer.Methods From March to November in 2009,605 women received cervical HPV testing in the Tumor Hospital of Harbin Medical University,to obtain specimens of cervical cytology,rapid flow-through hybridization technique (namely Hybribio flow-through hybridization)was used to detect HPV genotypes simultaneously.Single-factor and multivariate factors non-conditional Logistic regression analytic method was used to discuss the relationship between HPV infection of females and age,marital condition,level of education,level of income,occupation,initial age for sex,contraception,number of pregnancies,delivery approach and smoking.Results HPV infection rate was 21.49%(130/605),the positive rate of HPV infection in high-risk subtypes was 15.70%(95/605),the most common type was 5.29%(32/605)in the samples.Single factor non-conditional logistic regression model analysis showed that initial age for sex was the risk factor(X2=4.4618,P<0.05),HPV prevalence increased with a lower initial age for.sex reduced.But there was no significant difference in age,marital condition,education,income,occupation,contraception,number of pregnancy,delivery approach and smoking teams(X2=0.0525,1.8510,1.0348,0.2592,1.1176,1.5664,2.8835,1.4597,2.6161,all P>0.05).The analysis of multivariate factors nonconditional Logistic regression showed that the age of initially having sex,marital status and number of pregnancies were the risk factors(X2=21.6637,8.0574,15.7573,all P<0.05).Conclusions The risk factors for HPV infection are mainly about having sex too young,marital status and number of pregnancies,attention should be paid to screening for HPV.

Objective To design,synthesize and screen high efficient small interfering RNA(siRNA) targeting to cyclooxygenase-2(COX-2)on rheumatoid arthritis synovial fibroblasts(RASF).To further study the effect of specific COX-2 siRNA interfering on mediators of inflammatory cytokines.Methods Four pairs of siRNA for human COX-2 mRNA were synthesized by utilizing RNA design software,while another random sequence was designed as control.They were divided into group A to H.Among them,group A was used as the negative control(CTL),and group B to F were transfected as random siRNA(NC),1#～4#siRNA in order. These siRNAs were transferred into RASF by LipofectAMINE2000 package and PMA(phorbol-12-myristate- 13-acetate)was added into each culture and with a final concentration of 100 nmol/l.RASF was collected 48 hours after transfection.The expression of hCOX-2 at mRNA level was determined by reverse transcription- polymerase chain reaction(RT-PCR)and hCOX-2 protein level by Western Blot.The supernatant levels of PGE_2,IL-1?,IL-6,TNF-?and vascular endothelial growth factor(VEGF)of the above groups were detected by ELISA.Results The levels of hCOX mRNA and protein in RASF treated with 4-#siRNA were significantly lower than those of the negative control and other groups.The level of PGE_2 and cytokines like IL-1?,IL-6, TNF-?and VEGF in the supernatant were lower in the 4#siRNA group than in other groups.Conclusion 4#siRNA can effectively inhibit the expression of COX-2 mRNA and the synthesis of the COX-2 protein in human synovial fibroblasts.The level of PGE_2,IL-1?,IL-6,TNF-?and VEGF is the lowest in the super- natant.Thus 4#siRNA has been confirmed to specifically block the COX-2 in human synovial fibroblasts.

Objective To investigate the killer cell lg-like receptors (KIR) gene frequency of HIV-1 infected slow progressors(SP) and typical progressors(TP), and to analyze the interaction between KIR alleles and the progression of HIV-1 infection in Chinese population. Methods Eighty-one HIV-1 posi-tive individuals including 43 SPs and 38 TPs were recruited. Carriage of KIR genes was assessed using poly-merase chain reaction sequence-specific primers (PCR-SSP) assays. Results KIR2DS3 gene frequency was significantly lower in SP group (3.6%) than that in TP group (14.2%), P =0. 018 ,OR =0. 210,95% CI =0.053-0.833. The number of activating KIR genes was less in SP group than that in TP group, but was not significant (P = 0. 208). Conclusion Lower KIR2DS3 gene frequency may potentially be associated with slower progression to AIDS in Chinese population.

OBJECTIVE To analyze trimethylation of genome-wide histone H3 lysine 4(H3K4met3) induced by silicon dioxide(SiO2)through chromatin immunoprecipitation linked to microarrays(ChIP-chip)in lung fibroblast(LF)of rats. METHODS A primary co-culture model of rat alveolar macrophages (AM)and LF in vitro. AM were exposed to 100 mg · L-1 free SiO2 for 24 h,before LF were collected and the phenotype of LF was determined after transdifferentiation by immunohistochemistry. ChIP-chip was used to profile the variations of trimethylation in H3K4 of lung fibroblasts in CpG island regions. ChIP-qPCR was used to validate the microarray results. The mRNA expression of nfib and kpna3 was analyzed by qRT-PCR. RESULTS Totally 1815 (518 increased and 1297 decreased) genes of H3K4met3 displayed significant differences in SiO2 100 mg·L-1 group compared with control group(Cy3/Cy5 value>2.0 or <0.5,NimbleScan V2.5 software). The results of ChIP-qPCR were quite consistent with those of microarray. CONCLUSION There are significant differences in methylation of genome-wide H3K4 between SiO2 100 mg·L-1 group and control group. These novel candidate genes may become potential biomarkers or new interfered targets.

This study investigated the effect of puerarin on human umbilical vein endothelial cells (HUVEC) injured with hydrogen peroxide (H₂O₂). HUVEC were divided into three groups: a control group, a model group (H₂O₂ 400 μmol·L^-1) and a puerarin-treated group (3, 10, 30 and 100 μmol·L^-1). HUVEC were cultured with varied concentration of puerarin for 2 h and treated with H₂O₂ for another 24 h. Cell proliferation was detected by a CCK-8 assay. The mitochondrial membrane potential was measured by a JC-1 fluorescent probe. A transwell chamber assay was adopted to observe cell migration ability. Mitochondrial respiratory function was measured in a two-chamber titration injection respirometer (Oxygraph-2k). The expression of interleukin-1β (IL-1β), interleukin-18 (IL-18) and tumor necrosis factor-α (TNF-α) was detected by quantitative real-time PCR. The expression of pyroptosis-mediated proteins, including cleaved-cysteinyl aspartate-specific proteinase-1 (caspase-1), N-gasdermin D (N-GSDMD), NOD-like receptor protein 3 (NLRP3) and purinergic ligand-gated ion channel 7 receptor (P2X7R) was detected by Western blot. The results show that 400 μmol·L^-1 H₂O₂ treatment for 24 h causes obvious damage to HUVEC. Compared with the model group, puerarin protected against cellular injury in a dose-dependent manner, with the greatest effect at a dose of 30 and 100 μmol·L^-1. Puerarin significantly decreased the mitochondrial membrane potential and improved mitochondrial function. Puerarin inhibited cell migration induced by H₂O₂, suppressed the expression of IL-1β, IL-18 and TNF-α, and down-regulated the pyroptosis-mediated protein. These changes are statistically significant (P < 0.05). These findings demonstrate that puerarin has a protective effect against H₂O₂-induced oxidative damage of HUVEC by inhibiting the migration of HUVEC cells. The mechanism may be related to improved mitochondrial respiratory function and inhibition of pyroptosis.

Background and Objectives: The search for a suitable alternative for urethral defect is a challenge in the field of urethral tissue engineering. Induced pluripotent stem cells (iPSCs) possess multipotential for differentiation. The in vitro derivation of urothelial cells from mouse-iPSCs (miPSCs) has thus far not been reported. The purpose of this study was to establish an efficient and robust differentiation protocol for the differentiation of miPSCs into urothelial cells. Methods: and Results: Our protocol made the visualization of differentiation processes of a 2-step approach possible. We firstly induced miPSCs into posterior definitive endoderm (DE) with glycogen synthase kinase-3β (GSK3β) inhibitor and Activin A. We investigated the optimal conditions for DE differentiation with GSK3β inhibitor treatment by varying the treatment time and concentration. Differentiation into urothelial cells, was directed with all-trans retinoic acid (ATRA) and recombinant mouse fibroblast growth factor-10 (FGF-10). Specific markers expressed at each stage of differentiation were validated by flow cytometry, quantitative real-time polymerase chain reaction (qRT-PCR) assay, immunofluorescence staining, and western blotting Assay. The miPSC-derived urothelial cells were successfully in expressed urothelial cell marker genes, proteins, and normal microscopic architecture. Conclusions We built a model of directed differentiation of miPSCs into urothelial cells, which may provide the evi-dence for a regenerative potential of miPSCs in preclinical animal studies.

This study aimed to evaluate the vasorelaxant effect and mechanisms of compound reserpine and triamterene tablets (CRTTs) and its component triamterene on isolated rat thoracic aorta rings. Isolated rat thoracic aorta rings pre-contracted by high potassium or norepinephrine (NE) were used to evaluate the vasodilatory effect of CRTTs and its component triamterene. The mechanisms concerning endothelium, potassium channels and calcium channels were studied through the interventions of several tool drugs. Animal welfare and experimental procedures followed the requirements of the Laboratory Animal Management and Animal Welfare Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College. The results showed that both CRTTs and triamterene had potent relaxant effect on KCl and NE pre-contracted vessels. Triamterene showed partial endothelium dependency, and N-nitro-L-argininemethyl ester hydrochloride (L-NAME) could influence the relaxant effect, which may be related to the opening of calcium-activated potassium channels. Meanwhile, CRTTs could inhibit intracellular Ca²⁺ release and extracellular Ca²⁺ influx. Overall, CRTTs and its component triamterene have vasorelaxant effects on vessels in vitro, and the vasorelaxant effect of CRTTs may be related to intracellular Ca²⁺ release and extracellular Ca²⁺ influx, while this effect of triamterene may depend on vascular endothelial function and may also be related to the opening of calcium-activated potassium channels.

Objective To investigate the association between APOBEC3G mRNA levels in peripheral blood mononuclear cells(PBMCs)of HIV/AIDS patients and disease progression in Henan province.Methods Real-time PCR was used to detect the mRNA levels of APOBEC3G in PBMCs of HIV/AIDS patients at difierent disease progression stage.Flow cytometry and automated viral load analyzer were used to count CD4+ T cells and plasma HIV viral loads.Results The mRNA levels of APOBEC3G in HIV/AIDS patients were lower than in HIV-negative controls(t=4.887,P＜0.01),and APOBEC3G levels were significantly higher in slow development group than those in HIV and AIDS groups(P＜0.05).The levels of APOBEC3G mRNA correlated positively with CD4+ T cell counts(R2=0.190,P=0.002)and negatively with HIV-1 viral loads(R2=0.094.P=0.038).Conclusions The APOBEC3G mRNA levels in PBMC of HIV infected individuals are associated with HIV disease pmgression. Higher mRNA expression levels of APOBEC3G may be one of the protective factors which can play important role in delaying disease progression.