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 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 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 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.

Objective To assess the causes of sildenafil failure and the feasibility of successfully rechallenging non-responding patients. Methods A total of 126 consecutive erectile dysfunction (ED) patients from Jan 2009 to Dec 2009 who claimed poor response to sildenafil (sildenafil 100 mg on demand, at lease 4 epiodes) were enrolled into the study. All patients received sexual reeducation and were treated with sildenafil, taken on a daily dose of 50 mg for 4 weeks. The International Index of Erectile Function-5 (IIEF-5), Rigiscan, serum testosterone or penile Doppler were used to evaluate ED and the cause of on-demand sildenafil failure. End point efficacy of rechallenging was evaluated using the IIEF-5 and the sexual encounter profile (SEP) 'Were you able to insert your penis into your partner's vagina?' and 'Did your erection last long enough to achieve successful intercourse?'. Results The recruited patients comprised of 41 cases with psychological ED, 39 cases with hypogonadism ED, 28 cases with diabetes mellitus ED and 18 cases with vascular ED. Compared with pretreatment and on-demand sildenafil baseline, daily administration of sildenafil significantly enhanced all efficacy outcome variables. The IIEF-5 was significantly improved after daily sildenafil (12.3 ± 2.9 vs18. 8±4.4, P＜0.01), 78 patients responded to daily sildenafil. The overall salvage rate was 61.9%(78/126). Conclusions Sexual reeducation and daily administration of sildenafil may be able to salvage many patients with ED who were sildenafil non-responders.

Objective To estabalish a new conception, Erectile Dysfunction-no sexual life (ED-NS), so as create an evaluating questionnaire, Self-estimation Index of erectile function-No sexual life (SIEF-NS) so as to investigate its clinical reliability. Methods The conception of ED-NS was identified and the SIEF-NS questionnaire was established. Patients who complained of ED-NS and normal controls were enrolled into the research and assessed the erectile function was assessed with SIEF-NS.The SIEF-NS includes 12 questions, such as sexual libido, general erectile function, nocturnal penile erection, erectile function during foreplay, erectile function during audio-video sexual stimulation,confidence, depression, etc and each question has 5 point scales. Results Sixty-one ED-NS patients and 57 controls were enrolled into the study and assessed erectile function with SIEF-NS. The mean score of each question and integral score of SIEF-NS in ED-NS patients were significantly different from normal controls (P＜0. 05). When the integral score was 35 points according to the ROC curve of integral score, the sensitivity of SIEF-NS was 88.5% and specificity was 96. 5%. Conclusions ED-NS is a new conception to define patients who have erectile dysfunction without sexual life. SIEFNS is suggested to be a useful method for the evaluation of ED-NS patients.

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.