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 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 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 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 investigate the effects of exogenous BMSCs transplantation at different time points on the treatment of traumatic brain injury in rats. Methods: Male 3-week-old SD rats were taken,BMSCs was isolated and cultured in vitro to 3 genera-tions. Fifty healthy female SD rats were randomly divided into 5 groups,10 rats in each group:control group,brain trauma model group, early treatment group,middle treatment group,late treatment group. A traumatic brain injury model was made by using cortical trauma instrument in rats,after the establishment,the treatment group:early treatment group(1 day),middle treatment group(28 days),late treatment group model(56 days),were given BMSCs (1×106ml-1) intervention treatment,the control group and model group were given equal dose of normal saline. In addition,40 rats were tested by water maze,and each group was divided into four points:6 d,7 d, 8 d and 9 d after injury,the spatial memory ability of rats was detected. All the experimental groups were sacrificed at 14 days after treatment. Morphological changes of brain tissue were observed by HE staining;the expression of LC-3 and Beclin-1 protein was detected by Western blot. Results: The brain trauma model group was compared with the control group,the shape of neuronal cell bodies appears to change,a clear gap was seen between the cells,and necrotic neurons appeared. Compared with the brain trauma model group,the morphological changes of neurons were alleviated,and the expression of protein was obviously weakened in the early stage of traumatic brain injury (1 d) and the medium term group (28 d) (P＜0. 05). BMSCs was not effective in the treatment of advanced traumatic brain injury (56 d). Conclusion: In the early and middle stage of traumatic brain injury in rats,the transplantation of BMSCs may alleviate the extent of traumatic brain injury to some extent,and the effect of late treatment is not obvious.

Objective:To investigate the effect of bone marrow mesenchymal stem cells transplantation on the cognitive function in hippocampus after traumatic brain injury in rats.Methods: BMSCs were isolated and cultured from male 3-week-old SD rats in vitro.60 female SD rats were randomly divided into 3 groups(n=20),control group,model group,BMSCs treatment group.After the es-tablishment of brain trauma model of the model 1,7,14,28 d,BMSCs treatment group was given BMSCs(1×106ml-1)intervention.In addition,20 rats were tested by water maze,and each group was divided into four points:6,7,8 9 d after injury.The morphological changes of brain tissue was detected by HE staining;the water content of brain tissue was detected by dry wet method;Morris detection method,detection of spatial learning and memory ability in rats;nerve motor function score, neural function analysis detection;field test,detection of spatial learning and memory ability in rats.Results: The morphological changes of brain were observed by HE staining;the water content of brain tissue was detected by wet and dry gravity;the water maze test was conducted to examine the spatial learning ability,the motor function score,and the field experiment.The results of wet and dry gravity test showed that compared with the control group,the model group was significantly higher than the control group(P< 0.05),the water content of the treatment group at each time was significantly lower than the model group;the results of water maze test showed that the latency of safe island was significantly increased in the model group on the eighth and ninth day after injury,compared with the control group,and the difference was statistically significant(P<0.05).In the treatment group,the safe island latency was significantly shorter in the eighth day and ninth day after injury than in the control group;the neurological score results showed that compared with control group,model group,rats in the hind limb grasping reflex,vibrissae evoked forelimb positioning and lateral pad step tests were not normal performance,resulting in functional score decreased with statistical significance(P<0.05).The functional scores of the treatment group and the model group were significantly higher than those of the model group.Conclusion:Through the behavioral related tests,the data show that BMSCs can restore the neurological function to a certain extent after traumatic brain injury in rats.

AlM:To observe the changes of corneal endothelium after phacoemulsification cataract surgery in different types of cataract patients.
METHODS: Randomly selected age-related cataract, diabetic cataract and cataract of high myopia 30 eyes of 30 cases, respectively, in our hospital. All patients underwent phacoemulsification combined with intraocular lens implantation, corneal endothelial density and the percentage of hexagonal cells were measured by corneal endothelial cell instrument without touching before surgery and one week after surgery.
RESULTS: The difference of the preoperative corneal endothelial cell density and the percentage of hexagonal cells among three groups were not statistically significant (P>0. 05). One week after surgery, the cell density in three groups were respectively 2 496. 86 ± 298. 96/mm2 , 2 379. 51 ± 375. 13/mm2 , 2 425. 38 ± 312. 68/mm2 , the percentage of hexagonal cells were respectively ( 46. 20 ± 12. 03)%, (43. 44±13. 99)%, (44. 35±8. 13)%. Both the cell density and the percentage of hexagonal cells one week after surgery were lower than those before operation. There were significant difference in three groups ( P <0. 05). Both the measurements in diabetic cataract group and cataract of high myopia group after surgery were lower than those in age-related cataract group, the cell density and the percentage of hexagonal cells in diabetic cataract group were lower obviously compared with those in age- related cataract group and the difference was significant (P<0. 05).
CONCLUSlON:The tolerance of corneal endothelial cell to phacoemulsification cataract surgery is lower in cataract with diabetes and high myopia. Corneal endothelium should be assessed preoperatively and protected intraoperatively.

AIM To investigate the effect of Icarrin on penile erection and cyclic GMP levels in rabbit corpus cavernosum(RCC). METHODS Effects of various concentration of Icariin on cGMP level in RCC were measured by 125 I radioimmunoassay procedure with Sildenafil and papaverine as the positive controls RESULTS In the presence of SNP, Icariin increased cGMP concentration in rabbit corpus cavernosum in a dose dependent manner ( P