0.05). ConclusionThe expression of VOCC mRNA and BKCa mRNA in kidney tissues of IgA nephropathy patients are abnormal.There is positive correlation between the abnormal expression of VOCC mRNA and BKCa mRNA and total glomerular pathological lesions integrals.The expression of VOCC mRNA and BKCa mRNA in kidney tissues of IgA nephropathy may serve as the indictor for the disease progression.

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.
DNA/isolation & purification* ; DNA Fingerprinting ; Female ; Genotype ; Humans ; Male ; Polymerase Chain Reaction ; Silicon ; Spermatozoa

Hepatocyte growth factor (HGF) pretreatment could protect multiple cell types from apoptosis induced by various damages including oxidative stress. The present study was designed to investigate the protective effect of HGF on rat cortical neurons against apoptosis induced by hydrogen peroxide (H2O2) in culture, and then to explore whether HGF could influence the mitochondrial pathway of apoptosis. Primary rat cortical neurons were isolated from Sprague-Dawley rats and cultured in serum free medium containing 2% B27 and Neurobasal-A. To mimic the oxidative stress damage, cortical neurons were exposed to 100 mumol/L H2O2 for 4 h. To explore the effects of HGF on the neurons subjected to H2O2 injury, cells were pretreated with HGF 15, 30, 60 ng/mL for 24 h, respectively, and then exposed to 100 mumol/L H2O2 for 4 h. The cell viability was measured by MTT colorimetric assay and cell injury was evaluated by lactate dehydrogenase (LDH) leakage rate. Apoptotic cells were detected by Hoechst 33258 staining and Annexin V-FITC/PI double labeled flow cytometry. The caspase-3 activity was assessed by colorimetry. The alteration of transmembrane potential of mitochondria was determined by confocal laser scanning microscopy. The expression of cytochrome C protein was measured by Western blot analysis. The results showed that H2O2 treatment significantly decreased the cell viability, increased LDH leakage rate and the percentage of apoptotic cells. Pretreatment of HGF at different concentrations (15-60 ng/mL) could remarkably increase the cell viability of neurons. Compared with that of H2O2 group (53.4%+/-7.4%), the cell viabilities of neurons treated with 15, 30, and 60 ng/mL HGF significantly increased to (69.3+/-6.4)%, (77.5+/-6.1)% and (82.9+/-9.3)% (P<0.05), respectively. HGF preincubation also evidently decreased the LDH leakage rate in cortical neurons damaged by H2O2. The results of Hoechst staining revealed that HGF pretreatment could significantly reduce the apoptotic rate of neurons. The apoptotic rate of H2O2 group was (62.8+/-7.1)%, while that of HGF groups decreased significantly to (34.8+/-8.4)%, (23.5+/-3.2)% and (18.6+/-4.5)% (P<0.05), respectively. The data from caspase-3 activity assay indicated that HGF preconditioning could also remarkably decrease the caspase-3 activity of neurons. In addition, in the presence of various concentrations of HGF, the decrease of transmembrane potential of mitochondria in neurons caused by H2O2 injury could be reversed. Moreover, as detected by Western blot analysis, HGF downregulated the expression of cytochrome C protein in neurons. These results suggest that HGF has a protective effect on rat cortical neurons against apoptosis induced by H2O2, which might be related to the inhibition of the mitochondrial apoptotic pathway and the suppression of the caspase-3 activity.
Animals ; Apoptosis ; Brain ; cytology ; Caspase 3 ; metabolism ; Cell Survival ; Cells, Cultured ; Cytochromes c ; metabolism ; Hepatocyte Growth Factor ; pharmacology ; Hydrogen Peroxide ; pharmacology ; Mitochondria ; physiology ; Neurons ; cytology ; drug effects ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley

BACKGROUNDThe CRF07_BC recombinant strain has been one of the most predominantly circulated HIV-1 strains in China, it is therefore necessary and urgent to develop a relevant animal model to evaluate candidate vaccines targeting HIV-1 CRF07_BC. A highly replication-competent simian/human immunodeficiency viruses (SHIV) construct containing the Chinese CRF07_BC HIV-1 env gene with the ability to infect Chinese rhesus monkeys would serve as an important tool in the development of HIV vaccines. The aim of this study was to examine whether SHIV XJDC6431 with the env fragment from a Chinese HIV-1 isolate virus could infect the human and monkey peripheral blood mononuclear cell (PBMC), establish infection in Chinese rhesus macaque.

METHODSA SHIV strain was constructed by replacing the rev/env genes of SHIV KB9 with the corresponding fragment derived from the HIV-1 CRF07_BC strain. The infectious activity of the SHIV clones was determined in vitro in PBMCs from both non-human primate animals and humans. Finally, one Chinese rhesus macaques (Macaca mulatta) was infected with one SHIV via intravenous infusion.

RESULTSOne SHIV clone designated as SHIV XJDC6431, was generated that could infect macaque and human PBMC. The virus produced from this clone also efficiently infected the CCR5-expressing GHOST cell lines, indicating that it uses CCR5 as its coreceptor. Finally, the virus was intravenously inoculated into one Chinese rhesus macaque. Eventually, the animal became infected as shown by the occurrence of viremia within 3 of infection. The viral load reached 105 copies of viral RNA per ml of plasma during the acute phase of infection and lasted for 10 weeks post infection.

CONCLUSIONSWe conclude that SHIV XJDC6431 is an R5-tropic chimeric virus, which can establish infection not only in vitro but also in vivo in the Chinese rhesus macaque. Although the animal inoculated with SHIV XJDC6431 became infected without developing a pathologic phenotype, the virus efficiently replicated with a persistent level of viral load in the plasma. This suggested that the SHIV could be used as a tool to test candidate AIDS vaccines targeting the Chinese HIV-1 CRF_07BC recombinant strain.

Animals ; Chimera ; Genes, env ; HIV-1 ; genetics ; physiology ; Humans ; Macaca mulatta ; Proviruses ; genetics ; Receptors, CCR5 ; physiology ; Simian Immunodeficiency Virus ; genetics ; physiology

Objective To evaluate the application of “OmniView”, a new three-dimensional ultrasound technology, in displaying the fetal palate. Methods The three-dimensional volume data was acquired from 100 normal fetuses, analysed by OmniView technology with the facial midsagittal plane as the starting plane. The imaging of fetal palate was obtained in axial plane (through maxilla, oral cleft), coronal plane, oblique coronal plane (through piriform aperture, oral cleft, submental triangle), and the palate′s curved plane tiled imaging by drawing the anatomical lines on referenced sagittal plane (facial midsagittal plane). The volumes of ifve fetuses with cleft lip and palate were obtained and analysed by the same technology. Results The volume dataset of 91 (91.0%, 91/100) normal fetuses were acquired successfully, and analyzed by OmniView technology, the results of 91 normal fetal palate in different plane were: (1) In axial plane through maxilla, the visualization of alveolar process bow was 91 (100%, 91/91). It was shown as“C”shaped arcuate structure, the anechoic structure of alveolar socket could be seen on the bow, and the ifrst 6 alveolar sockets were displayed clearly. The visualization number of hard palate was 91 (100%, 91/91), it was shown as hyperechoic lfake between two sides of alveolar bones. In axial plane through oral cleft, the visualization number of soft palate was 81 (89.0%, 81/91), it was shown as a strip of soft tissue echo band. (2) In coronal plane, the visualization number of hard palate was 91 (100%, 91/91), it was shown as a strip of hyperechoic band and separated the oral and nasal cavity. (3) In oblique coronal plane through piriform aperture, the visualization number of hard palate was 91 (100%, 91/91), it was shown as a short strip of hyperechoic band. In oblique coronal plane through oral cleft, the visualization number of hard palate was 91 (100%, 91/91). In oblique coronal plane through submental triangle, the visualization number of hard palate was 91 (100%, 91/91). In the above two planes, the hard palate was shown as a strip of hyperechoic band, due to acoustic shadow behind the hard palate, the nasal cavity and nasal septum above the hard palate couldn’t be displayed. (4) In oblique coronal plane through piriform aperture, the visualization number of soft palate was 81 (89.0%, 81/91). The visualization number of uvula was 25 (27.5%, 25/91). The soft palate was shown as a lfake of soft tissue echo behind the hard palate, and the uvula was shown as papillary protrusions on the edge of the soft palate in the midline. In oblique coronal plane through oral cleft, the visualization number of soft palate was 81 (89.0%, 81/91). In oblique coronal plane through submental triangle, the visualization number of soft palate was 81 (89.0%, 81/91). In the above two planes, the soft palate was shown as a strip of soft tissue echo band, the soft tissue echo of fetal tongue was in the lower front of soft palate, and the anechoic region of nasopharynx was superior behind the soft palate. (5) In the curved plane tiled imaging of palate, the visualization number of alveolar process bow (primary palate) was 91 (100%, 91/91). The visualization number of hard palate was 91 (100%, 91/91). The visualization number of soft palate was 81 (89.0%, 81/91). the visualization number of uvula was 25 (27.5%, 25/91), the planar panorama of alveolar process bow, hard palate and soft palate could be visualized intuitively, the alveolar arch and hard palate were shown as bone-like hyperecho, and the soft palate was shown as soft tissue hypoecho. In iffteen cases′volume involved cleft lip and palate, all five cases of malformations were detected through three-dimensional data analysis, the position and range of the cleft palate could also be conifrm. Abnormal fetuses were all veriifed after induction of labor. Conclusions By three-dimensional ultrasound technology-“OmniView”, the axial and coronal plane of fetal palate could be obtained easily which was dififcult by two-dimensional ultrasound, and the special oblique coronal plane of secondary palate could be displayed easily. The panorama of the palate could be visualized intuitively though curved plane tiled imaging by drawing a line tracking the structure of the palate. This technology could simplify the ultrasound examination procedure of the fetal palate, reduce the operators′skill-dependence, and quickly evaluated the integrity of the fetal primary palate and secondary palate. For the cleft lip fetus, this technology can determine whether the cleft palate exist or not, together with their position and range.

Objective To explore informatized management of medical consumables in new situation.Methods lntormatized management of medical consumables was executed with the involvement of multi-dimension data analysis,medical consumables supervision and approval system under early warning mechanism,secondary development of sunshine purchase platform as well as big data rebuilding and etc.Results The whole-course supervision from purchase to utilization was realized for medical consumables,which provided data support to medical consumables utilization.Conclusion Wholecourse,professional and informatized management is implemented for medical consumables,which lays a foundation for clinical service and hospital transformation.

OBJECTIVEChimeric human/bovine immunodeficiency virus (HBIV) cDNA was constructed by replacing HIV tat and LTR with bovine immunodeficiency virus (BIV) tat and LTR to study the activity of BIV tat and LTR in the chimerae.

METHODSThe target fragments of BIV tat, LTR and HIV gag, pol, env were respectively amplified by using PCR and sequentially inserted into pBluescript SK(+) vector. The chimeric clone was transfected into human MT4 cells. The transcript and gene expression of the HBIV chimeric virus were detected by using RT-PCR and a reverse transcriptase assay, respectively.

RESULTSBIV tat mRNA and HIV gag mRNA were detected. The reverse transcriptase activity of the chimeric virus was analyzed in the fluctuation curve.

CONCLUSIONSIn chimeric HBIV cDNA transfected MT?4 cells, BIV tat and HIV gag were transcripted. The reverse transcriptase of the chimeric virus had biological activity. These data suggest that in MT4 cells, BIV LTR had promoter activity and BIV tat had the function of transactivation in the chimeric virus. The study of the chimeric virus with infectivity is in progress.

AIDS Vaccines ; Animals ; Cattle ; Cloning, Molecular ; DNA, Complementary ; genetics ; DNA, Viral ; genetics ; Genes, gag ; genetics ; Genes, pol ; genetics ; Genes, tat ; genetics ; HIV-1 ; genetics ; Humans ; Immunodeficiency Virus, Bovine ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transcription, Genetic ; Transcriptional Activation ; Transfection ; Virus Replication

BACKGROUNDSome larger scale, randomized studies have demonstrated the superiority of drug-eluting stents (DES) over bare metal stents (BMS) for the treatment of acute myocardial infarction (AMI). This study aimed to investigate the impact of DES, in comparison with BMS, on the 2-year clinical outcomes in patients with ST-elevation myocardial infarction (STEMI).

METHODSFrom January 2002 to December 2008, a total of 1301 consecutive STEMI patients treated with coronary stenting in Shenyang Northern Hospital were prospectively registered. Patients received BMS (n = 868) or DES (n = 435) implantation in the infarction related artery according to physician's discretion. A propensity score analysis was performed and two well matched subgroups were selected (BMS, n = 288; DES, n = 288) to evaluate the 2-year clinical outcomes. The primary outcome was the occurrence of major adverse cardiac events (MACE), which was defined as a composite of all-cause death, myocardial infarction (MI), or target vessel revascularization (TVR).

RESULTSSurvival salvage analysis showed that 2-year cumulative hazards were not significantly different between the two groups with respect to TVR (2.8% vs. 3.1%, log-rank P = 0.780), stent thrombosis (1.7% vs. 4.2%, log-rank P = 0.079) and MACE (8% vs. 10.8%, log-rank P = 0.236). Multivariate analysis showed that DES was an independent protective factor of MI (HR: 0.211, 95%CI: 0.049 to 0.908) and stent thrombosis (HR: 0.327, 95%CI: 0.107 to 0.994).

CONCLUSIONDES was associated with similar 2-year clinical outcomes to those of BMS for the treatment of STEMI in daily practice.

Aged ; Angioplasty, Balloon, Coronary ; methods ; Drug-Eluting Stents ; adverse effects ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; mortality ; therapy ; Prospective Studies ; Stents ; adverse effects ; Thrombosis ; etiology ; Treatment Outcome

OBJECTIVETo evaluate the long-term outcomes of successful or failed revascularization in patients with chronic total occlusions (CTO).

METHODSThe clinical data of 1332 consecutive patients underwent percutaneous coronary intervention (PCI) for CTO between June 1993 and December 2006 in our hospital were analyzed. These patients were divided into two groups according to the procedural success (n = 1202) or failure (n = 130).

RESULTSOverall success rate of procedure was 90.2% (1202/1332). The patients in CTO success group experienced a superior 10-year survival rate (76.9% vs. 64.6%, log rank P = 0.012) and a significantly higher no major adverse cardiovascular event (MACE) survival rate (41.8% vs. 27.6%, log rank P < 0.001) compared to the patients in CTO failure group. During the long-term follow-up, the proportion of patients who accepted coronary artery bypass grafting (CABG) was significantly lower in CTO success group than that in the CTO failure group (4.3% vs. 14.6%, P < 0.001).

CONCLUSIONSuccessful PCI procedure leads to increased long-term survival and MACE-free survival and the reduced need for CABG for patients with CTO lesions.

Aged ; Angioplasty, Balloon, Coronary ; Arteriosclerosis Obliterans ; therapy ; Coronary Occlusion ; therapy ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; Treatment Outcome

BACKGROUNDThere are limited data on the efficacy of drug-eluting stents (DES) for treatment of chronic total occlusions (CTO). The aim of the study was to evaluate the long-term clinical outcomes of DES implantation for CTO compared with bare-metal stent (BMS) implantation.

METHODSBetween June 1995 and December 2006, a total of 1184 patients with successful recanalization of at least one de novo CTO lesion were consecutively registered, including 660 (55.7%) who underwent DES and 524 (44.3%) who underwent BMS implantation. All patients were followed up for up to 5 years for occurrence of major adverse cardiac events (MACE). Long-term survival rates were estimated with the Kaplan-Meier method.

RESULTSBaseline clinical and angiographic characteristics were comparable between the two groups except that patients in the DES group received longer dual antiplatelet therapy ((7.4 +/- 2.5) months vs (1.7 +/- 0.8) months, P < 0.001). Average follow-up periods were (4.7 +/- 0.89) and (3.2 +/- 1.3) years for the BMS and DES groups, respectively. There was no significant difference in 5-year survival rates between the two groups (90.3% for DES group vs 89.6% for BMS group, Log-rank P = 0.38), but the 5-year target vessel revascularization (TVR)-free survival rate in the DES group was significantly higher than that in the BMS group (81.6% vs 73.5%, Log-rank P < 0.001). The cumulative MACE-free survival in the DES group was also significantly higher than that in the BMS group (80.6% vs 71.5%, Log-rank P < 0.001). The rates of re-admission caused by cardiovascular disease (27.0% vs 37.8%, P < 0.001) and the need for bypass surgery were significantly lower in the DES group (1.5% vs 3.4%, P < 0.05). By multivariable analysis, DES implantation could significantly lower the long-term MACE risk of PCI for CTO patients (HR: 0.492; 95% CI 0.396 - 0.656, P < 0.001). Left ventricular ejection fraction < 50% and elderly (> or = 65 years) were identified as independent predictors of long-term MACE during follow-up.

CONCLUSIONThis study demonstrates the long-term (up to 5 years) efficacy of DES for treatment of CTO, which is superior to BMS implantation in reducing the rates of TVR and MACE, as well as the need of re-admission and bypass surgery.

Aged ; Angioplasty, Balloon, Coronary ; methods ; Coronary Angiography ; Coronary Occlusion ; therapy ; Drug-Eluting Stents ; adverse effects ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Proportional Hazards Models ; Stents ; adverse effects ; Treatment Outcome