ABSTRACT
In order to establish a real-time RT-PCR based on SYBR Green Ⅱ for detection of hepatitis E virus (HEV),a pair of special primers was designed according to the conserved sequences of ORF2 in GenBank.Result showed that the standard curve of established SYBR Green Ⅱ real-time RT-PCR had a wide dynamic range from 4.10 × 102-4.10 × 108 copies/μL with a linear correlation(r2) of 0.996.The sensitivity could reach 1.00 × 102 copies/μL.The melting curve analysis using SYBR Green Ⅱ dye showed one specific peak with a melting temperature(Tm) of 84.0 C ±0.1 C.No amplification was detected from the RNA samples of porcine reproductive and respiratory syndrome virus,classial swine fever virus,transmissible gastroenteritis virus,porcine bocavirus,porcine epidemic dearrhoea virus porcine kobuvirus and porcine rotavirus by this PCR,respectively.Excellent reproducibility was obtained for detecting constructed positive plasmid DNA with intra-assay of 0.83 ％-0.94 ％ and inter-assay of 0.83％-0.94％.Further detection of 61 specimens showed that 9 of them were HEV positive,and the results of the quantitative RT-PCR were the same as that of the conventional RT-PCR.In conclusion,the real-time quantitative RT-PCR for HEV is feasible,the real-time RT-PCR established in this study will be useful for earlier rapid laboratory diagnosis and pathogenesis of HEV.
ABSTRACT
<p><b>BACKGROUND</b>Generally, intracytoplasmic sperm injection (ICSI) may be the preferable method to treat partial globozoospermia, but whether there exist some correlations between ICSI fertilization rate and the proportion of round-headed sperm or morphologically normal sperm remains open. This study was to explore the correlation between ICSI fertilization rate and the sperm morphology in patients with partial globozoospermia.</p><p><b>METHODS</b>Thirty-four patients diagnosed with partial globozoospermia accepted the following assisted fertilization treatments - 2 cases accepted in-vitro fertilization (IVF) alone, 26 cases accepted ICSI alone, and 6 accepted split IVF/ICSI. Detailed morphological characteristics were described using Diff-Quik rapid staining. Sixty cases accepting IVF or ICSI treatment in our reproductive center were considered as the control group after being matched by relevant criteria. Fertilization rate, embryo quality, embryo implantation rate and clinical pregnancy rate were calculated.</p><p><b>RESULTS</b>Besides very high proportion of round-headed sperm, partial globozoospermia also showed very high proportion of small-acrosomal sperm and very low proportion of morphologically normal sperm. Fertilization rate of IVF (IVF alone plus split IVF) was very low in partial globozoospermia (25.4% ± 17.4%), but ICSI (ICSI alone plus split ICSI) achieved satisfying fertilization rate compared with the control group (66.2% ± 22.5% vs. 68.8% ± 29.4%, P > 0.05). In patients with partial globozoospermia, there were no correlations between ICSI fertilization rate and the proportion of round-headed sperm, small-acrosomal sperm, or morphologically normal sperm.</p><p><b>CONCLUSIONS</b>There was high proportion of small-acrosomal sperm in partial globozoospermia. For patients with partial globozoospermia, ICSI is more preferable than IVF. ICSI fertilization rate does not depend on the proportion of round-headed sperm, small-acrosomal sperm, or morphologically normal sperm.</p>
Subject(s)
Adult , Female , Humans , Male , Pregnancy , Fertilization in Vitro , Infertility, Male , Therapeutics , Sperm Injections, Intracytoplasmic , Spermatozoa , Congenital Abnormalities , Cell Biology , PhysiologyABSTRACT
Objective To study the peripheral neutrophils activation mesenteric lymph in a murine hemorrhagic shock model.Methods In this study,18 male Sprague-Dawley rats were evenly divided into 3 groups.Group A:rats subjected to hemorrhagic-shock and Ringer's lactate(RL)resuscitation,group B:rats suffered from no blood loss but received same amount of RL as in group A,and group C:rats experience no blood loss nor RL transfusion.The main mesenteric lymphatic duct was cannulated with 24G catheter in all rats.In group A,blood was withdrawn through femoral artery until mean arterial pressure reached 40?5 mm Hg,the pressure was maintained for 90 min.In group B,no blood was withdrawn,these two groups received RL 3 times as the blood withdrawn,in group C,no blood was withdrawn,nor fluid was given.Lymph samples during pre-shock,the first hour and second hour post-shock or sham shock were collected and was used to induce PMN activation.Mesenteric lymph-induced rat PMN(polymorphonuclear neutrophil)adhesion molecule expression(CD18 and CD11b)and neutrophil respiratory burst activity was examined using a FACS flow cytometer.Results In group A,1st hour and 2 nd hour post-shock mesenteric lymph induced rat PMN activation,the expression of CD11b was 63.28?1.13%,61.23? 1.16%,respectively,compared with control(P