Extraction of sperm DNA from mixed stain by the modified differential lysis method combined with silicon bead method / 法医学杂志

OBJECTIVE@#To extract sperm DNA from mixed stain by the modified differential lysis method combined with silicon bead method and to evaluate its application value.@*METHODS@#Fifty-two mixed stains containing female STR genotypes detected by differential lysis method were collected. The sperm DNA was extracted by the modified method combined with silicon bead method, then genotyped with the Identifiler Kit, and compared with the results of genotyping by the conventional differential lysis method as control.@*RESULTS@#Of the 52 samples, 38 samples with sole male STR genotypes in all loci were detected. The detection rate of male STR genotypes was 98.08% through the modified method combined with silicon bead method.@*CONCLUSION@#The modified differential lysis method combined with silicon bead method can be used in extraction of sperm DNA from mixed stain.

Effect of fusion protein TAT and heme oxygenase-1 on liver sinusoidal endothelial cells apoptosis during preservation injury / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Proteins or peptides can be directly transferred into cells when covalently linked to protein transduction domains (PTDs). TAT is one of the most widely studied PTDs. The effect of fusion protein TAT and heme oxygenase-1 (HO-1) on liver sinusoidal endothelial cells (SECs) apoptosis during cold storage is unknown. The present study aimed to determine whether fusion protein TAT-HO-1 would transduce efficiently into liver during cold storage, and, if so, to determine whether TAT-HO-1 would attenuate SECs apoptosis during preservation injury in rat.</p><p><b>METHODS</b>Livers of Sprague-Dawley rats were harvested and randomly assigned to group 1 (HTK solution) and group 2 (HTK solution containing TAT-HO-1 fusion protein) according to the type of the preservation solution. The transduction efficiency of TAT-HO-1 was examined and the impairment of SECs was assessed during the period of cold storage followed by 1 hour of reperfusion.</p><p><b>RESULTS</b>TAT-HO-1 can transduce efficiently into liver during cold storage. A significantly lower apoptotic index of SECs was observed in group 2, at 6, 12 and 18 hours of cold storage after 1 hour reperfusion, when compared with group 1. TAT-HO-1 reduced HA and ET levels in liver at each time point. Both Bcl-2 and Bax protein were expressed in hepatocytes and SECs at the periphery of the sinusoidal space. Moreover, higher Bcl-2 expression and lower Bax expression were observed in group 2.</p><p><b>CONCLUSIONS</b>TAT-HO-1 can transduce efficiently into rat livers and shows a protective effect on SECs by attenuating apoptosis during cold ischemia/reperfusion injury. Protein transduction will be a novel therapeutic strategy to reduce the risk of preservation injury in liver transplantation.</p>

Protection of human islets from induction of apoptosis and improved islet function with HO-1 gene transduction / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Islet transplantation represents an ideal therapeutic approach for treatment of type 1 diabetes but islet function and regeneration may be influenced by necrosis or apoptosis induced by oxidative stress and other insults. Heme oxygenase-1 (HO-1) is the rate-limiting enzyme in the catabolism of heme into biliverdin, releasing free iron and carbon monoxide. It has also been reported to be an antioxidant enzyme which can improve the function of grafted islets by cytoprotection via free radical scavenging and apoptosis prevention. In the present study, we investigated whether transduction of HO-1 genes into human islets with an adenovirus vector has cytoprotective action on islets cultured in vitro and discuss this method of gene therapy for clinical islet transplantation.</p><p><b>METHODS</b>Cadaveric pancreatic islets were isolated and purified in vitro. Transduction efficiency of islets was determined by infecting islets with adenovirus vector containing the enhanced green fluorescent protein gene (Ad-EGFP) at multiplicities of infection (MOI) of 2, 5, 10, or 20. Newly isolated islets were divided into three groups: EGFP group, islets transduced with Ad-EGFP using MOI = 20; HO-1 group, transduced with adenovirus vectors containing the human HO-1 gene using MOI = 20; and control group, mock transduced islets. Insulin release after glucose stimulation of the cell lines was determined by a radioimmunoassay kit and the stimulation index was calculated. Flow cytometry was used to detect apoptotic cells in the HO-1 group and in the control group after induction by recombinant human tumor necrosis factor-alpha (rTNFalpha) and cycloheximide (CHX) for 48 hours.</p><p><b>RESULTS</b>Adenovirus vectors have a high efficiency of gene transduction into adult islet cells. Transduction of islets with the Ad-EGFP was most successful at MOI 20, at which MOI fluorescence was very intense on day 7 after transduction and EGFP was expressed in cultured islet cells for more than four weeks in vitro. The insulin release in the control group was (182.36 +/- 58.96) mIU/L after stimulation by high glucose media (16.7 mmol/L), while insulin release from the HO-1 group and the EGFP group were (270.09 +/- 89.37) mIU/L and (175.95 +/- 75.05) mIU/L respectively. Compared to the control group and the EGFP group, insulin release in the HO-1 group increased significantly (P < 0.05). After treatment with rTNFalpha and CHX the apoptotic ratio of islet cells was (63.09 +/- 10.86)% in the HO-1 group, significantly lower than (90.86 +/- 11.25)% in the control group (P < 0.05).</p><p><b>CONCLUSIONS</b>Transduction of human islets with Ad-HO-1 can protect against TNF-alpha and CHX mediated cytotoxicity. The HO-1 gene also appears to facilitate insulin release from human islets. Transduction of donor islets with the adenovirus vector containing an HO-1 gene might have potential value in clinical islet transplantation.</p>

Association between endothelial nitric oxide synthase gene (G894T) polymorphism and essential hypertension in Uygur populations / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the association between G894T polymorphism and essential hypertension (EH) in Uygur population in Xinjiang province.</p><p><b>METHODS</b>In this case-control study, G894T genotypes in 375 hypertension patients (EH group) and 414 normotensive control subjects (NT group) from the rural area of Tuluafan of Xinjiang was investigated by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.</p><p><b>RESULTS</b>(1) Significant differences were found in GG, GT and TT frequencies of G894T genotypes between the EH and NT groups (56.5%, 28.3%, 15.2% in EH group and 65.9%, 22.5%, 11.6% in NT group, OR = 2.97, 95% CI 1.393 - 6.358). T allele frequencies were significantly higher in EH group (29.33%) than that in NT group (22.83%, P < 0.05). (2) SBP, DBP in patients with T allele of eNOS gene [(171.36 +/- 22.30) mm Hg and (103.63 +/- 13.22) mm Hg] were significantly higher than that of GG genotype [(158.07 +/- 20.850) mm Hg and (89.90 +/- 10.39) mm Hg] (P < 0.01).</p><p><b>CONCLUSIONS</b>eNOS gene exon7 G894T polymorphism might relate to essential hypertension in Uygur population in Xinjiang province.</p>