Study on the association of USP8 gene polymorphisms with male infertility in ethnic Han Chinese from Sichuan / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the association of single nucleotide polymorphisms (SNPs) of ubiquitin-specific protease 8 gene (USP8) with male infertility among ethnic Han Chinese from Sichuan.</p><p><b>METHODS</b>A total of 316 infertile males were recruited (case group), which included 72 severe oligozoospermic (SO) cases and 244 non-obstructive azoospermic (NOA) cases. The control group consisted of 149 fertile males. The genotypes of 4 SNPs (rs2241769, rs11857513, rs7174015 and rs3743044) were determined with a Sequenom MassArray technique. The frequencies of genotype, allele and haploptye were analyzed.</p><p><b>RESULTS</b>No significant difference was detected in the allelic or genotypic frequencies of the 4 SNPs between the two groups (P>0.05). Based on linkage disequilibrium analysis and haplotype construction, the frequency distribution of haplotype CAAG showed a significant difference between non-obstructive azoospermic patients and the controls (P=0.021).</p><p><b>CONCLUSION</b>The 4 SNPs (rs2241769, rs11857513, rs7174015 and rs3743044) of USP8 gene may not be associated with male infertility in ethnic Hans from Sichuan. While the haplotype CAAG may be a down-regulating factor for the risk of NOA.</p>

Study on in vitro activities of sulfamethoxazole combined with trimethoprim against Mycobacterium tuberculosis / 国际检验医学杂志

Objective To evaluate the bacteriostasis of sulfamethoxazole(SMZ)combined with trimethoprim(TMP)against My‐cobacterium tuberculosis(MTB) in vitro ,so as to provide basis for clinical application .Methods The minimal inhibitory concentra‐tion(MIC) value of TMP/SMZ against MTB ,including standard sensitive strain(H37Rv) ,clinical sensitive strains(20 strains) and clinical multiple‐drug‐resistance strains(MDR ,30 strains) ,was detected by using MPP observation method .Results The MIC val‐ue of standard strain H37Rv was TNP(0 .5 μg/mL)+SMZ(9 .5 μg/mL) .The growth of 40 strains(accounted for 80% ) of clinical isolates ,including 17 sensitive strains and 23 MDR strains ,could be inhibited by TMP(1 μg/mL)+ SMZ(19 μg/mL) compound . Conclusion TMP combined with SMZ may has good antibacterial activity for strains of MTB in vitro .

The diagnostic value of T-SPOT.TB and ADA for tuberculous pleurisy / 国际检验医学杂志

Objective To evaluate the diagnostic value of peripheral blood T-SPOT.TB and pleural effusion′s adenosine deami-nase(ADA)activity in patients with tuberculous pleurisy.Methods Sixty-two patients with suspected tuberculous pleurisy were enrolled in the study,whose peripheral blood T-SPOT.TB and pleural effusion ADA were measured and the best diagnostic thresh-old was determined by ROC curve.Results According to the diagnostic criteria of tuberculous pleurisy,24 patients were diagnosed with tuberculous pleurisy,33 were non-tuberculous pleurisy and 5 undiagnosed in the end.The sensitivity of T-SPOT.TB for the diagnosis of tuberculous pleurisy was 91.7 %,the specificity was 81.8%,Positive predictive value was 78.6%,negative predictive value was 93.1%.The ADA activity was (40.5±15.4)IU/L in tuberculous pleurisy group,which was higher than in non tubercu-lous pleurisy group[(2.4 ±9.5 )IU/L](P <0.01 ).The cut-off value of ADA was 22.5 IU/L in tuberculous pleurisy diagnosis, while its sensitivity and specificity were 83.3% and 84.8% respectively.The sensitivity of the combined detection of T-SPOT.TB and ADA was 95.8%.Conclusion The combined detection of peripheral blood T-SPOT.TB and pleural effusion ADA has a higher sensi-tivity in diagnosis of tuberculous pleurisy,and has important auxiliary diagnostic value for patients with suspected tuberculous pleurisy.

Association of single nucleotide polymorphisms in H2BFWT with male infertility in southwest China / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the association of single nucleotide polymorphisms (SNPs) in exons of H2BFWT gene with male infertility in southwest China.</p><p><b>METHODS</b>Three hundred and twelve infertile men and 211 fertile men were recruited. PCR was employed to amplify the target fragments of H2BFWT, and PCR products were sequenced. Prevalence of SNPs in the two groups was analyzed by statistical method.</p><p><b>RESULTS</b>The detected SNPs have mainly distributed in the first exon of the H2BFWT gene. The ratios of 368G/A (rs553509) and -9C/T (rs7885967) were significantly higher in infertile group than fertile group. Additionally, a context-dependent effect was observed between 368G/A and -9C/T which the allele 368G combined with allele -9T will considerably increase the risk of male infertility.</p><p><b>CONCLUSION</b>The present study has revealed that the SNPs in H2BFWT are associated with male infertility, and may increase the susceptibility of male infertility in southwest China.</p>

Analysis of human papillomavirus 16 subtypes E2 gene and long control region sequence in Chengdu / 中华传染病杂志

Objective To explore the relevance between sequence variation of human papillomavirus (HPV)16 subtypes E2 gene or long control region (LCR) and cervical lesions.Methods Fifty specimens from HPV16 infected people in Chengdu were collected,including cervical exfoliated cells from 38 HPV carriers,papilloma tissues from 8 cases of genital warts,2 with cervical intraepithelial neoplasia (CIN) Ⅱ and 2 with CIN Ⅲ in this study.Polymerase chain reaction was used to amplify E2 gene and LCR,then an evolutionary tree was constructed.Results In all the 50 specimens,there were 12 mutation sites in E2 gene,among which,C→A existed in one specimen of genital warts,and ≥2 mutation sites existed in all the other 48 specimens.There were 28 mutation sites of LCR sequence of all the specimens.Ten specimens were chosen to construct evolutionary tree and were sequenced.The data showed that 8 specimens were Asian variants,E2 gene mutation existed in all the specimens while the LCR 7867 G→A only existed in the four CIN.Conclusion LCR 7867G→A is a correlative mutation site of cervical lesions in Chengdu.

Rapid detection of SOX2 gene by primed in situ labeling / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To rapidly detect SOX2 gene using primed in situ labeling (PRINS).</p><p><b>METHODS</b>Human peripheral blood samples were cultured using an optimized method. Sequence of the SOX2 gene was amplified in situ with biotin-labeled specific primers and processed with a tyramide signal amplification (TSA) biotin system. Subsequently, fluorescence-stained signal was detected by streptavidin-Texas red. For the control group, MCF-10F cells were transfected with Lentivirus hSox2.</p><p><b>RESULTS</b>By VideoTesT-FISH software analysis, the long arm of chromosome 3 in the experimental group showed a specific red fluorescence signal, whilst the control samples showed no specific signals for SOX2. Transfected MCF-10F cells showed various efficiency of SOX2 gene integration.</p><p><b>CONCLUSION</b>PRINS utilizes a highly sensitive in situ PCR technique combined with fluorescence labeled oligodeoxynucleotides can synthesize probes in situ, thus greatly reducing the cost of probe and time for detection. It can facilitate identification and classification of induced pluripotent stem cells, and has many potential applications in this prospect.</p>

Size distribution of cell-free DNA in maternal plasma / 国际检验医学杂志

Objective To investigate the size distribution and change of plasma DNA in different gestational weeks between the normal and abnormal pregnancy.Methods Two methods were applied to measuring the lengths of different derived plasma DNA fragments.For the maternal derived DNA,GAPDH gene distribution of each DNA in size-fractionated fragment was assayed by real-time PCR after gel electrophoresis separation;for the fetal derived DNA,size distribution of SRY gene of fetal derived DNA was also determined by using a series of real-time PCR.Results Compared with that of matenal derived DNA,the fragment length of fetal derived DNA was mostly shorter than 300 bp.Compared with that in mid-pregnancy,plasma maternal derived DNA fragment distribution had no significant difference in late-pregnancy and in patients with high-risk of carrying a Down's child,whereas fetal derived DNA fragment was a bit longer.Conclusion The fragment length of fetal derived DNA is significantly longer than that of fetal one in maternal blood.Moreover,because of abnormal pregnancy and accompanied with the pregnancy progress,the number and physico-chemical property(fragment length,for example) of fetal derived DNA has a certain change.

Detection of the SRY gene by primed in situ labeling / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish a primed in situ labeling (PRINS) technique which can be more effective in detection of single copy gene.</p><p><b>METHODS</b>On the basis of traditional PRINS, new reagents and procedures, such as TaqStart antibody, four primers of the sex determining region Y (SRY) gene and TSA(TM) Biotin System were included in detection of the SRY gene. Meanwhile, fluorescence in situ hybridization(FISH) to detect the SRY gene was used as control.</p><p><b>RESULTS</b>Fifty metaphases were scored. PRINS labeling showed signals for the SRY on the Y chromosome at band Yp11.3 in all metaphases. These signals were as distinct as that from results of FISH.</p><p><b>CONCLUSION</b>This improved method is ideal for rapidly localizing single copy genes and small DNA segments. And PRINS is a cost- and time-effective alternative to FISH.</p>