OBJECTIVE: To establish a 29 Y-STR loci multiplex PCR system for investigating the genetic polymorphisms and to assess its application value in forensic science. METHODS: A multiplex PCR system was established using a five color fluorescence labeling 29 Y-STR loci (DYS456, DYS389 I , DYS437, DYS447, DYS389 11, DYS438, DYS522, DYS460, DYS458, DYS622, DYS390, DYS392, DYS448, DYS449, DYS391, Y-GA TA-H4, DYS388, DYS19, DYS385a/b, DYS527a/b, DYS393, DYS459a/b, DYS635, DYS439, DYS570 and DYS627) for multiple amplification and capillary electrophoresis. And its applicability was validated with genetic polymorphism data of 29 Y-STR of unrelated 2,000 male samples in Shandong Han population. RESULTS: A total of 1,981 different haplotypes of 2,000 individuals showed genotype diver- sity between 0.370 0 and 0.965 4. The system provided stable and accurate typing with high sensitivity of 0.05 ng. It satisfied the needs of variety of routine biological samples. CONCLUSION The 29 Y-STR loci multiplex PCR system could be applied for actual cases and establishment of Y-STR database. In addition, it has great significance in forensic science practices and related research.
Asian People/genetics* ; China ; Chromosomes, Human, Y ; DNA/isolation & purification* ; Ethnicity/genetics* ; Forensic Genetics/methods* ; Forensic Sciences ; Genetics, Population/methods* ; Haplotypes ; Humans ; Male ; Microsatellite Repeats ; Multiplex Polymerase Chain Reaction/methods* ; Polymorphism, Genetic ; Reproducibility of Results

Objective To explore the repair possibilities of endothelial progenitor cells (EPCs)in peripheral blood in patients with different extents of obstructive sleep apnea (OSA) through measuring the levels of pro-angiogenic factors and different subgroups EPCs in peripheral blood in patients with OSA. Methods Ninety adult patients with OSA, 30 healthy controls with matched age and gender were enrolled for this study. The subjects performed Polysomnography, were divided in-to four group based on Apnea Hypopnea Index (AHI). The serum levels of HIF-1α, SDF-1αand VEGF were assessed by ELISA. Mononuclear cells were isolated from peripheral blood with density gradient centrifugation, and flow cytometry was used to detect levels of CD133+KDR+EPC, CD133+CD34+EPC, CD34+KDR+EPC and ALDHloCD34+KDR+EPC based on AL-DH activity, and CD133, CD34, PE-KDR related cell surface markers. Results The levels of CD133+KDR+EPC, CD133+CD34+EPC, CD34+KDR+EPC were higher in OSA groups than those of control group, both of which were higher in severe OSA group than those of in mild and moderate OSA groups. The levels of ALDHloCD34+KDR+EPC were higher in mild and moderate OSA groups than that of the control groups, and the levels of ALDHloCD34+KDR+EPC were significantly lower in se-vere OSA group than those of control, mild and moderate OSA groups. Serum levels of HIF-1α. VEGF were significantly high-er in OSA groups compared to those in control groups, both of which were higher in severe OSA group than those of mild and moderate OSA groups. Serum levels of SDF-1αwere significantly lower in severe OSA groups than those of mild, moderate OSA and control groups (P<0.05). Conclusion The mobilization and recruitment of different subtypes of EPCs are obvious-ly increased in patients with OSA, but ALDHloCD34+KDR+EPC with vascular repair capacity keeps to invariability, even de-creases in patients with severe OSA, which results in endothelial damage, and increases the risk of cardiovascular disease.

Objective To study the influence of HBV replication regulator,enhancer I and Pre- S2,on the immune response of HBV DNA vaccine.Methods DNA fragments of HBsAg,PreS2 HBsAg,HBsAg-enhancer I and PreS2-HBsAg-enhancer I region of HBV were amplified by PCR using the complete genome DNA of HBV adr subtype,and inserted into VR1012 vectors,respective- ly.The recombinant plasmids were transfected into HepG2 cells,and injected into Balb/C mice.The expression of HepG2 cells and the cellular and humoral immune response of mice were tested by cell immuno-chemistry,ELISA and ELISPOT.Results The target protein were expressed by transfected HepG2 cells,enhancer I and Pre-S2 can promote the expression of HBsAg in transfected cells.The HBsAb and the HBsAg specific CTL in inoculated mice were found in the second week after injection, PreS2 but not enhancer I can promote the immune response in inoculated mice.Conclusions When inserted into HBV DNA vaccine,enhancer I and PreS2 can promote the expression of HBsAg in transfected HepG2 cells,PreS2 can promote the immune response in inoculated Balb/C mice.

Objective To explore the mechanism of vessel endothelial dysfunction in rats under intermittent hypoxia (IH). Methods The respiratory simulation system was used to simulate IH. Sixty C57BL/6J rats (male) were randomized into control group and IH group. The rats of IH group were exposed to IH 8 hours per day for 6 weeks. The serum levels of hypoxia inducible factor (HIF)-1a and stromal cell derived factor (SDF)-1a were assessed by ELISA. The serum levels of reactive oxygen species (ROS) were detected in two groups. The serum expression of miR-199a-5p was detected by real-time fluorescent quantitative PCR in two groups. The dual luciferase report system and point mutation test were used to verify target gene for HIF-1a. Results The serum levels of HIF-1a and SDF-1a were significantly higher in IH group than those of control group (μg/L:1.60±0.02 vs. 1.19±0.02, 1 823.00±8.97 vs. 1 444.00±17.90, P<0.01). The serum level of ROS was significantly higher in IH group than that of control group (U/mL:487.66±35.73 vs. 211.57±23.82, P<0.01). The serum level of miR-199a-5p expression was significantly lower in IH group compared to that of control group (1.31±0.07 vs. 3.47± 0.17, P<0.01). The result of dual luciferase reporter gene detection confirmed that target gene of miR-199a-5p was HIF-1a. Conclusion The serum level of miR-199a-5p is decreased first due to IH, and then its target gene (HIF-1a) is increased. HIF-1a can induce the increased level of SDF-1a, and its receptor (CXCR-4 ) is also increased. Finally, HIF-1a can increase the serum level of ROS, resulting in the endothelial dysfunction.

Objective To compare safety and feasibility using radial versus femoral access during cardiac catheterization of patients who had previously undergone coronary artery bypass graft ( CABG) surgery. Methods We retrospectively evaluated 116 consecutive patients who underwent graft intervention via the transradial (TRA group, n = 46) or transfemoral approach (TFA group, n = 70), and observed the baseline clinical characteristics, angiography characteristics and complications between the 2 groups. Results The baseline clinical characteristics between the 2 groups were similar ( all P > 0. 05) . No significant difference was observed in angiography characteristics and procedural parameters including operation time, radiation exposure and puncture time between the 2 groups (all P > 0. 05). There was no significant difference in major adverse cardiac events during hospitalization. PCI to graft vessels were all successful and procedural success rates were similar between the 2 groups (P = 0. 669). Vascular access site complications were significantly lower ( P = 0. 03) in the TRA group. No access site complication was recorded in the TRA group. 7 cases (10. 0% ) with complications were recorded in the TFA group including 1 case of major bleeding (1. 4% ), 3 cases of minor bleeding (4. 3% ), 2 cases of local hematorna (2. 9% ) and 1 case of A-V fistula formation. Conclusions In contrast to the transfemoral route, the rate of major vascular complications was negligible using the transradial approach.

Objective To explore effect of physic temperature dropping by ice blanket on incidence of pressure ulcer of stroke patients in ICU, and normalize the basic nursing procedure during temperature reduc-tion.Methods 230 fever patients were randomly divided into the group A(control temperature with ice blan-ket,108 cases)and the group B (control temperature with routine measure,122 cases).Thc incidence of pressure ulcer and surface temperature of pressed position in the two groups were compared.Results The incidence of pressure ulcer and surface temperature of pressed position in group A was lower than those of group B.Conclusions The incidence of pressure ulcer can be reduced by using ice blanket for physic temperature dropping.The pressure of pressed position were descended by alternative prostration and lateral position.

OBJECTIVETo observe anti-atherosclerotic effect of Xuefu Zhuyu Granule (XZU) and Danlou Tablet (DT) on blood lipids, platelet derived growth factor (PDGF), vascular smooth muscle cells (VSMCs) proliferation, extracellular signal-regulated kinase (ERK) signal pathway in atherosclerosis (AS) model rats, and to explore their potential mechanisms.

METHODSForty male Wistar rats were randomly divided into five groups, i.e., the normal control group, the model group, the Atorvastatin group, the DT group, the XZG group, 8 in each group. Rats in the normal control group were fed with basic forage for 12 weeks, while rats in the other four groups were fed with high fat forage plus intraperitoneal injection of vitamin D3 to build AS model. Then rats in the model control group, the Atorvastatin group, the DT group, the XZG group were administered with normal saline, Atorvastatin suspension (0.18 mg/mL), DT suspension (45 mg/mL), and XZG (1 g/mL) by gastrogavage for 8 successive weeks, respectively. After intervention serum levels of TC, TG, LDL-C, HDL-C, and PDGF were detected by ELISA. Pathological changes in thoracic aorta were observed by HE staining. Protein expression levels of ERK1/2 and pERK1/2 in thoracic aorta were measured by Western blot.

RESULTSCompared with the normal group, serum TC, TG, LDL-C, PDGF levels, and expression levels of ERK1/2 and pERK1/2 significantly increased (P <0. 05) in the model control group. HE staining showed irregular intimal thickness, accumulated endothelial foam cells, lipids deposited, disarranged media VSMCs, forming typical AS plaque. Compared with the model group, TC and PDGF levels decreased in all medicated groups (P < 0.05, P < 0.01). Serum levels of TG and LDL-C significantly decreased in the Atorvastatin group and the DT group (P < 0.01, P < 0.05). Expressions of ERK1/2 and pERK1/2 significantly decreased in the Atorvastatin group, the DT group, and the XZG group (P < 0.01). HE staining also showed typical AS plaque in three medicated groups, but with reduced pathological degree of endometrial hyperplasia and plaque area.

CONCLUSIONSXZG and DT could reduce the plaque area and attenuate pathological degree of AS in model rats, thereby postponing the progress of AS. Its mechanism might be achieved through reducing serum lipids and release of PDGF, inhibiting ERK signal pathway activation and VSMC proliferation.

Animals ; Aorta, Thoracic ; Atherosclerosis ; drug therapy ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Extracellular Signal-Regulated MAP Kinases ; Lipids ; Male ; Plaque, Atherosclerotic ; Rats ; Rats, Wistar ; Tablets

ObjectiveTo assess the cause of tadalafil failure and the feasibility of successfully rechallenging nonresponding patients.Methods A total of 80 consecutive erectile dysfunction ( ED ) patients who claimed poor response to tadalafil were enrolled into the study.A self-administered tadalafil-use questionnaire composed of eight questions was applied to assess how they had used tadalafil.Subjects were given thorough instruction based on individual answers and four doses of tadalafil 20mg.After a 2-week follow-up,end point efficacy of rechallenge was evaluated using the sexual encounter profile (SEP),which was recommended by international advisory panel in 2004.ResultsA total of 45 subjects had one or more areas of major suboptimal use of tadalafil:21.2％ did not know that sexual stimulation was necessary for tadalafil to work,87.5％ attempted to use tadalafil less than four times,57.5％ took a maximal dose less than 20 mg,and 84％ felt nervous or anxious.Of the 65 patients undergoing tadalafil rechallenge,30 patients answered “yes” to SEP2 and SEP3.The response rate to rechallenge was 46.2％ ( 30/65 ).ConclusionsInappropriate use of tadalafil was major cause of tadalafil non-pesponse.The efficacy of tadalafil could be improved to a better extent by education of patients.

Objective To discuss the application value of transesophageal echocardiography (TEE) in occluding procedure with cardiopulmonary bypass (CPB) for children muscular ventricular septal defects (mVSD).Methods Forty-nine mVSD patients for occluding procedure with CPB were performed guided and monitored by TEE,9 cases were simple mVSD,40 were mVSD combined with other cardiac malformations.The location,size,number,scope and the surrounding anatomy of mVSD were carefully observed before going into cardiac arrest,preparing the operator to choose appropriate occluder devices.The procedure was evaluated after the recovery of heart beat using TEE.Results In all of the 49 patients underwent occluding procedure,44 cases were implanted into one occluder,5 implanted into two occluders.The results of TEE after operation showed that the positions of all occluders were not shifted,2 had trace residual shunt,there were no complications worsening,such as mitral and aortic regurgitation,etc.One patient abandoned treatment for serious infection,the other patients' occluders were all in normal position followed up at 6 months to 2 years after operation,there were no tiny residual shunt and valves regurgitation except 1 patient.Conclusions The technique that TEE guiding and monitoring in occluding procedure with cardiopulmonary bypass for children muscular ventricular septal defects can shorten the time of cardiopulmonary bypass,significantly reducing complications and risks of the surgery,improving the successful rate.

Objective To evaluate the efficacy of microvasoepididymostomy for the treatment of obstructive azoospermia. Methods Ninety-eight patients were confirmed as obstructive azoospermia by infertility investigations.The mean (range) age was 31 (20-43) years.The mean duration of obstruetion was 4 years.All patients were seen by the surgeon for a complete history and physical examination.Semen analyses proved azoopermia,serum levels of sexual hormone were normal and testicular function for production of sperms were normal certified by biopsy.All patients with suspected epididymal or vasal obstruction were offered scrotal exploration.The decision for microvasoepididymostomy was made during surgery,based on the pateney of the vas towards the abdomen documented by saline and sperm fragments containing in the epididymal fluid on intraoperative light-microscopic examination.Microvasoepididymostomy was performed in 58 patients documented epididymal obstruction The initial semen analysis was then done after usually 3 months postoperatively.Patency was arbitrarily defined as＞10 000 sperm/mL in ejaculate in at least one semen analysis after surgery.Postoperative patency rate and postoperative impregnation rate were followed. Results Fifty patients were followed up for 3-29 months,8 cases lost.Sperm was found by semen analysis in 36 patients.Sperm density was 4×104-2×108 sperms/ml and motility was 2%-70%.Semen analysis reveals azoospermia in 6 patients and the patients were counseled to undergo further testing to determine the ultimate outcome of the procedure.Natural conception occurred in 14 patients followed for more than 12 months.The overall pateney rate was 72%(36/50).Among patients with a follow-up of＞6 months,the natural paternity rate was 28%(14/50).The median time to achieve a natural pregnancy was 6.6 (4.0-10.0)months. Conclusion A more favourable patency can be achieved using microsurgical intussusception vasoepididymostomy and a part of patients with obstructive azoospermia can be cured by this means.