Intraflagellar transport is essential for spermiogenesis / 中华男科学杂志

Intra flagellar transport (IFT) is an evolutionarily conserved mechanism thought to be essential for the assembly and maintenance of most eukaryotic cilia and flagella. Development of the sperm tail axoneme resembles the cilia formation, which is organized by intraflagellar transport (IFT). Of all mammalian cells, sperm have the longest motile cilia, but few studies are reported on the role of IFT in the formation of sperm flagella and the mechanisms of IFT in spermiogenesis. This article focuses on the role of IFT in spermatogenesis and the importance of IFT in male fertility.

The role of SPAG6/SPINK2 protein complex in the formation of sperm acrosome in mice / 中华男科学杂志

Objective@#To explore the expression and regulatory function of sperm-associated antigen 6 (SPAG6) in the formation of the sperm acrosome in mice.@*METHODS@#The expression of SPAG6 during the first wave of spermatogenesis on postnatal days (PN) 8, 12, 16, 20, 24, 28, 30 and 35 was examined by Western blot and the localization of SPAG6 in the testicular germ cells was determined by immunofluorescence. The expression plasmids of SPAG6 and serine protease inhibitor Kazal-type 2 (SPINK2) were constructed, the interaction between SPAG6 and SPINK2 in the AH109 and CHO cells examined by yeast two-hybrid and co-localization assays, and the expression and localization of SPINK2 in the testicular germ cells of the SPAG6-knockout (SPAG6 KO) mice detected by immunofluorescence.@*RESULTS@#SPAG6 was highly expressed between PN 16 and 28 and localized in the acrosome of the round spermatids. Yeast two-hybrid assay showed the growth of SPAG6 and SPINK2 in the selective culture medium SD/-Leu/-Trp/-His, and the transfection of the CHO cells revealed the co-localization of SPAG6 and SPINK2 around the nuclei. The expression and acrosomal localization of SPINK2 were not found in the testicular germ cells of the SPAG6-KO mice.@*CONCLUSIONS@#SPAG6 interacts with SPINK2 and probably participates in the formation of the sperm acrosome by stabilizing the expression of SPINK2 during spermatogenesis.

Epidemiological survey and study of hepatitis E virus among HIV-infected population in Hubei Province / 中华疾病控制杂志

Objective To determinate the seroprevalence of hepatitis E virus (HEV) exposition among HIV-infected people in Hubei Province so as to provide basic data for effective prevention and control of HIV and hepatitis E virus． Methods A total of 335 serum samples of HIV-infected people were collected from January to June 2017．Serum samples were subjected to anti-HEV IgG andanti-HEV IgM screening. Data were statistically analyzed. Results In 335 HIV-infected people，0.89% (3/335) of serum samples were anti-HEV IgM positive and 41.49% (139/335) of them were anti-HEV IgG positive．The anti-HEV IgG positive rate was not linearly correlated with age ( 2linear=0.756，P=0.385), and there was no significant difference between the age groups (P>0.05). There was no significant difference in anti-HEV IgG positive rate between different gender( 2=0.085，P=0.771)．Anti-HEV IgG positive rate for the crowd with CD4 value ≤ 200 cell/μl was higher than that with CD4 ＞200 cell/μl，but showed no significant difference( 2=1.016，P=0.314)，and CD4 indexes of three positive anti-HEV IgM patients were 222, 446 and 198 in cell/μl．The anti-HEV IgG positive rate in HIV-infected population in eastern Hubei (30.77%, 32/1 041) was statistically different from that in central Hubei (47.92%, 46/96) ( 2=6.169，P=0.013) , and it was also different from that in southeastern Hubei (45.79%， 49/107) ( 2=5.034，P=0.025). Conclusions There were higher HEV infection rate among HIV-infected people in Hubei Province; thus, it is essential to positively carry out double-way screening for these two diseases at the same time, and effective prevention and control of these viruses are required.

Comparison of Efficacy and Safety of Clopidogrel and Ticagrelor in the Treatment of Acute non-ST Elevation Myocardial Infarction / 中国药房

OBJECTIVE:To compare clinical efficacy and safety of clopidogrel and ticaprelor in the treatment of acute non-ST elevation myocardial infarction (NSTEMI).METHODS:A total of 160 NSTEMI patients admited in cardiovascular disease department of our hospital during Oct.2013-Nov.2015 were divided into observation group and control group according to random number table,with 80 cases in each group.Both groups received routine treatment and continues intravenous pump of Tirofiban hydrochloride sodium chloride injection at 0.05 μg/(kg·h).Control group was additionally given oral loading-dose of Clopidogrel hydrogensultate tablet 300 mg,adjusted to 75 mg,qd,on the basis of routine treatment.Observation group was additionally given oral loading-dose of ticaprelor 180 mg,adjusted to 90 mg,bid,on the basis of routine treatment.Both groups received 1 month of treatment.The rate of platelet aggregation,LVEF,LVEDD,fibrinogen levels,the incidence of MACE and bleeding events were compared in 2 groups before and after treatment.RESULTS:Before treatment,there was no statistical significance in the rate of platelet aggregation between 2 groups (P＞0.05).After 1 week and 1 month of treatment,the rates of platelet aggregation in 2 groups were decreased significantly,and the observation group was significantly lower than the control group,with statistical significance (P＜0.05).After 1 week of treatment,there was no statistical significance in the levels of LVEF or LVEDD between 2 groups (P＞0.05).After 1 month of treatment,LVEF of observation group was significantly higher than that of control group,LVEDD was significantly lower than control group,with statistical significance (P＜0.05).There was no statistical significance in the level of fibrinogen before treatment and after 3 d of treatment (P＜0.05).After 1 week and 1 month of treatment,the levels of fibrinogen in 2 groups were decreased significantly,and the observation group was significantly lower than the control group,with statistical significance (P＜0.05).The incidence of MACE in observation group was 11.25％,which was significantly lower than 25.00％ of control group,with statistical significance (P＜0.05).There was no statistical significance in the incidence of bleeding (3.75％ vs.7.50％) between 2 groups (P＞0.05).CONCLUSIONS:Compared with clopidogrel,ticaprelor effectively inhibits platelet aggregation in NSETMI patients,reduces the level of fibrinogen,improves cardiac function and prognosis and doesn't increase the risk of bleeding with good safety.

Clinical value of N-terminal B-type natriuretic peptide precursor on auricular thrombosis in patients of coronary heart disease combined with atrial fibrillation / 中国医师进修杂志

Objective To analyze the predictive value of N-terminal B-type natriuretic peptide precursor(NT-proBNP) on auricular thrombosis in patients of coronary heart disease (CHD) combined with atrial fibrillation (AF).Methods The clinical data of 124 patients of CHD combined with AF were analyzed retrospectively.All patients were divided into research group (60 cases,compliance with auricular thrombosis) and control group (64 cases,non-compliance with auricular thrombosis),according to the occurrence of auricular thrombosis.The values of NT-proBNP,fasting plasma glucose (FPG),postprandial plasma glucose(2 h PG),total cholesterol (TC),high density lipoprotein cholesterol (HDL-C),oxidized low density lipoprotein (ox-LDL),high sensitivity C reactive protein (hs-CRP) and left ventricular ejection fraction (LVEF),left ventricular end diastolic diameter (LVEDd) and left atrial diameter (LAD) were detected and compared between two groups.The relative factors to the occurrence of auricular thrombosis were confirmed by multivariate Logistic analysis.The best cutoff point of NT-proBNP was confirmed by the areas under the receiver operating characteristic curve (ROC).Results The values of NT-proBNP,ox-LDL,hs-CRP,LVEDd and LAD in research group were higher than those in control group [(4 312.6 ± 209.1) pmol/L vs.(3 421.6 ± 156.8) pmol/L,(4.0 ± 0.9) mmol/L vs.(3.4 ± 0.8) mmol/L,(7.4 ± 1.3)mg/L vs.(5.8 ± 1.0) mg/L,(74.3 ± 6.8) mm vs.(58.1 ± 5.5) mm,(39.6 ± 4.3) mm vs.(32.5 ± 3.8) mm],LVEF and HDL-C were lower than those in control group [(48.2 ± 3.1)％ vs.(57.3 ± 3.8)％,(0.72 ± 0.16)mmol/L vs.(1.08 ±0.27) mmol/L],and there were significant differences (P ＜0.05).There were no significant differences in the values of FPG,2 h PG and TC between two groups (P ＞ 0.05).Multivariate Logistic analysis showed that NT-proBNP and LAD were independent factors to the occurrence of auricular thrombosis (P =0.009,0.028).There was significant difference in the occurrence of auricular thrombosis between patients with NT-proBNP ＞ 4 250 pmol/L and patients with NT-proBNP≤4 250 pmol/L (P =0.028).Conclusion NT-proBNP is an independent predictor for the occurrence of auricular thrombosis to patients of CHD combined with AF.

Cloning NS1 gene of H5N1 avian influenza virus and apoptosis induced by it in human pulmonary carcinoma cell line A549 / 病毒学报

The NS1 gene of the H5N1 subtype avian influenza virus was amplified by RT-PCR, and the am-plified product was cloned into the eukaryotic expression vector pCMV-Myc, then it was transfected into A549 cells. After 48 h, the expression of NS1 was detected by Western blot. Fluorescence and electron microscopy and flow cytometry showed that the NS1 gene of H5N1 avian influenza virus could induce apop-tosis in human pulmonary carcinoma cell line A549.

Construction, expression and preliminary pharmacokinetic analysis of IL-1ra mutants / 生物工程学报

Interleukin-1 receptor antagonist (IL-1ra), a member of IL-1 family, is a naturally occurring IL-1 inhibitor as "receptor antagonist", which blocks biological responses mediated by IL-1. Recombinant human IL-1ra (rhIL-1ra, Kineret) was introduced in clinical trials involving patients with RA. Between 2001 to approximately 2002, rhIL-1 ra was approved by the US Food and Drug Administration and the European Agency for the Evaluation of Medicine Procedure. Unfortunately, 10,000 to 100,000-fold excess amounts of IL-1ra are needed to relieve disease because minimal IL-1 can induce complete biological responses, and the dosage of 100 to approximately 150mg/day in a RA patient is so big that it greatly influence patients' physical, psychological and economical situation. In this study, IL-1ra mutants were established by site-specific mutagenesis to improve its stability. The sites of mutagenesis included R6 K7-AA,R93 K94-AA and K97 R98-AA. IL-1ra and its mutants were expressed in E. coli BL21 (DE3) using pTIG-Trx expressing system with the induction of IPTG. The recombinant proteins were purified by Ni2+ chelate chromatography and Sephadex G75 gel filtration chromatography. The activity of mutants is as high as IL-1ra. We characterized the pharmacokinetic profile of IL-1ra and its mutants. The third mutant's half life is 2.26 times than wt IL-1ra. The study has provided some approaches and experience for further research to improve the metabolism stability of IL-1ra.

Soluble expression and target study to brain of anti-TfR ScFv / 生物工程学报

The single-chain antibody gene (ox26-scFv) to transferrin receptor (TfR) was synthesized and amplified by three-step PCR. After sequencing, the gene was cloned into prokaryotic expression vector pTIG-Trx which carried thioredoxin (Trx) gene and a C-terminal His.tag. The Ox26-scFv proteins achieved 31% yields of total bacteria proteins at 20 degrees C, after 0.02mM IPTG induction using the strain E. coli BL21 (DE3). The soluble scFv proteins in cytoplasm suspension were about 35% and the inclusion bodies were about 65%. The soluble products were purified by immobilized metal chelation affinity chromatography (Ni-NTA), a single band with molecular weight 29 kD appeared on SDS-PAGE gel. Rat GH3 cell immunocytochemistry staining showed that Ox26-scFv protein could recognize and bind to transferrin receptor. Injected SD rats with Ox26-scFv proteins by tail veins, the antibodies were detected from brain tissues specially on the brain capillaries 4 h later which indicate that Ox26-scFv proteins have a good target function to brain capillaries and can permeate the blood-brain barrier mediated by the transferrin receptors.