Objective To explore the effect of oxidative stress on human Wiskott-Aldrich syndrome related protein 2 (WAVE2) expression in placental trophoblasts in women with preeclampsia.Methods (1) Twenty women with preeclampsia and twenty-three normal term pregnant women,delivered from August 15,2011 to February 23,2012 in the First Affiliated Hospital of Chongqing Medical University,were recruited and divided into preeclampsia group and control group.Placenta samples were collected after cesarean section.The localization and distribution of WAVE2 in placenta was studied by immunohistochemistry.Quantitative real-time polymerase chain reaction and Western blot were employed to assay the WAVE2 mRNA and protein levels.Tissue homogenates was applied to determine the levels of reactive oxygen species (ROS).The correlation between ROS levels and WAVE2 was also analyzed.(2) An in vitro hypoxia/reoxygenation (H/R) model was utilized to simulate ischemia/reperfusion injury to placental trophoblasts.The HTR-8/ SVneo cells (immortalized human first trimester extravillous trophoblast cells) were pre-incubated overnight,after exposure to H/R or normoxic conditions for 48 hours.Flow cytometry was employed to analyze intracellular ROS level.Meanwhile,Transwell assay was utilized to analyze the invasion and migration of HTR-8/SVneo cells.The location and expression of WAVE2 in trophoblasts was evaluated by cell immunofluorescence and Western blot.Statistical differences between the two groups were evaluated by independent t-tests.Pearson's correlation coefficient test was used for correlation analysis.Results (1)Compared with the control group,preeclampsia group had significantly higher 24-hour proteinuria [(1.96±0.24) g vs (0.08±0.05) g,t=19.436,P＜0.05],systolic blood pressure [(154 ± 13) mm Hg vs (98 ±11) mm Hg,t=11.324,P＜0.05] and diastolic blood pressure [(105±14) mm Hgvs (69±8) mm Hg,t=9.612,P＜0.05].In addition,the placental weight and birth weight of infants in preeclampsia group were significantly reduced as compared to the control group [(432±53) g vs (536±67) g,(2446± 187) g vs (3207± 233) g,t=14.562 and 16.307,allP＜0.05)].The WAVE2 mRNA level (0.28±0.07 vs 1.01±0.02,t=12.747,P＜0.05) and the WAVE2 protein levels (0.63±0.08 vs 1.34±0.19,t=11.648,P＜0.05) were also significantly decreased in preeclampsia groups.The level of ROS in placenta in the preeclampsia group was significantly higher than in control group [(144.22 ± 12.32) nmol/(mg · prot) vs (75.17 ± 8.71) nmol/(mg · prot),t=20.467,P＜0.05].There was significant negative correlation between ROS level and WAVE2 protein expression in preeclamptic placenta (r =-0.726,P =0.000).(2) In vitro study showed that,the levels of ROS in normoxia group and H/R group was (82.9±5.8)％ and (155.6±8.1)％,(t=12.747,P＜0.05).Compared with normoxia condition,decreased cell invasion and migration were found in HTR-8/SVneo cells in H/R group [(51.9 ± 3.3)％ and (58.4 ±4.2)％ respectively,t=11.034 and 13.839,P＜0.05].Results from the cell immunofluorescence showed that WAVE2 protein located in the cytoplasm of HTR-8/SVneo cells,and the expression of WAVE2 protein was significant decreased in HTR-8/SVneo cells after exposure to H/R for 48 h (0.37±0.05 vs 0.76±0.06,t=8.631,P＜0.05).Conclusions Excessive oxidative stress in preeclamptic placentas was correlated with the decreased expression of WAVE2.H/R-induced oxidative stress could decrease WAVE2 expression,which may contribute to impaired trophoblast invasion and migration in preeclampsia.

The effects of FeSO_4 upon the physiologicproperties were studied with isolated guinea pig a-trium. The FeSO_4 0. 2mmol/L, makes the contrac-tility of the atrium muscle decrease from 100% ofcontrol to 67 ? 9%. The concentration ofepinephrine inducing automaticity increased andthe duration-intensity curve was shifted to theright. The functional refractory period (FRP) wasprolonged from 298 ? 26 ms to 402 ? 43 ms. Thenegative inotropism and the inhibition of the auto-maticity suggest that FeSO_4 may influence Ca~(2+)movement, while the excitability of suppressionand prolongation of FRP indicate that it may bedue to its inhibition of Na~+ influx.

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

The study aims to solve the instability problem of methylphenidate (MPH) in plasma, and establish a LC-MS/MS method for simultaneous determining of MPH in human plasma. The stabilities of MPH in different media were studied, and the degradation characteristics of MPH in these media were also investigated by HPLC and LC-MS/MS. To a 200 microL aliquot of freshly collected plasma sample, 10 microL 2% formic acid was added immediately to prevent the hydrolysis of MPH in human plasma samples. Chromatographic separation was performed on a Sapphire C18 column using the mobile phase of methanol - 5 mmol.L-1 ammonium acetate buffer solution containing 0.1% formic acid (46 : 54). MPH was quantified by tandem mass spectrometry operating in positive electrospray ionization mode with multiple reaction monitoring. The detection used the transitions of protonated molecules at m/z 234.2-->84.1 for MPH and m/z 260.3-->183.1 for propranolol (IS), separately. The intra- and inter-assay precisions were all below 5.0%. The accuracies were all in standard ranges. The linear calibration curve was obtained in the concentration range of 0.035-40 ng.mL-1. The methods fulfilled the demand. The method was used to determine the concentration of MPH in human plasma after a single dose of 36 mg MPH tablet to 6 healthy Chinese volunteers. The method is suitable for the precisely determination of MPH and for pharmacokinetic study of MPH in human plasma.

Objective To evaluate the value of fecal high mobility group box-1 (HMGB 1 ) in early diagnosis and prediction of the severity of neonatal necrotizing enterocolitis (NEC). Methods From July 2013 to June 2015 , the neonates who had distention, vomit, or gross blood in stool and were suspected of NEC were recruited as NEC group while hospitalized children without abdominal distension, vomiting, bloody diarrhea, or other gastrointestinal symptoms were recruited as the control group. Stool samples were collected on day 1 , 3 , 5 and 7 after admission. The level of HMGB 1 was measured by enzyme linked immunosorbent assay (ELISA). Results In the end, there were 46 cases in NEC group and 15 cases in control group. In NEC group, 29 cases were conifrmed of stageⅠof NEC by abdominal radiograph within 24 h after hospitalized, all of them were deteriorated to stageⅡphase in 4 days, and 10 cases were deteriorated to stageⅢ. Seventeen cases were conifrmed of stageⅡby abdominal radiograph within 24 h after hospitalized, 7 cases were deteriorated to stageⅢ. In 17 stageⅢcases, 11 cases received surgical treatment and 6 cases gave up. Eight cases survived and 3 died after surgery. The levels of HMGB 1 in NEC group on day 1 , 3 , 5 and 7 after hospitalized were higher than those in control group (P

Objective To establish and assess the high-birth-weight offspring model of the diabetic rat induced by stueptozotocin,and the long-term metabolic impact of maternal hyperglycemia of those offsprings.Methods Streptozotocin (STZ,25 mg/kg) was given to Wistar rats (G group,n =14) once intraperitoneally to induce maternal hyperglycemia model (blood glucose between 10-20 mmol/L),and there still had a number of rats defined as severe hyperglycemia model group (SG group,n =5).The Control group (C group,n =7) were given the same volume citrate buffer solution.The body weight and blood glucose were recorded,and the lavaging glucose tolerance test (LGTT) was performed by a glucose meter in the gestation.The offsprings were corresponding allocated into 2 groups,and the birth weight were recorded.All the offsprings were observated body weight,blood glucose blood pressure (male rats only),and so on.Results (1) The blood glucose of G group (16.8 ±5.4 mmol/L) and SG group (20.5 ±5.6 mmol/L) were increased significantly as compared with C group (7.0 ± 1.4 mmol/L) 5 days after the model was established (P ＜ 0.01) ; and the average blood glucose of G group (16.6 ± 3.4 mmol/L) and SG group (23.8 ± 1.5 mmol/L) increased too as comparede with C group (5.8 ± 1.1 mmol/L),the difference was significance according to statistics (P ＜ 0.01).(2) According to the LGTT result,which operationed on generation day 4 and day 10,the blood glucose of every time point of G group were increased significantly as compared with C group (P ＜ 0.01).(3) The male and female birth weight of G group was remarkably higher than the C group and the SG group (P ＜ 0.05),and the blood glucose of SG /G/C group was (6.5 ±1.2) mmol/L,(4.1 ± 0.8) mmol/L,(4.1 ± 0.8) mmol/L respectively,according to the statistics results,the difference between SG group and G/C group respectively both remarkable (P ＜ 0.05).(4) The body weight,Lee's index,fat weight,and the fat weight of mass ratio in C group mother rats after lactation presented dressed compared with the SG group (P ＜ 0.05),and so as to the G group compared with the SG group (P ＜ 0.05).(5) In the female offsprings of G group,the birth weight was remarkably increased compared with the C group (P ＜ 0.05) ; the body weight of the female offsprings presented an increased trend compared with the C group since the 12 weeks,but had no statistical significance; there were significant differences of body weight between G group and C group since 15 weeks (P ＜ 0.05),and the trend kept up until 26 weeks; in the male offsprings of G group,the body weight on birth day and 4 weeks had a marked rise compared with the C group (P ＜ 0.05) ; and from then on,the body weight of the male offsprings presented an increased trend compared with the C group,but had no statistical significance until 26 weeks (P＞0.05).(6) In G group,the blood glucose on 30 min and 60 min of LGTT in female offsprings were increased than the C group since 20 weeks (P ＜ 0.05) ; the blood glucose of LGTT (30 min) still had a marked rise until 24 weeks (P ＜ 0.05) ; in G group,the blood glucose on 30 min of LGTT in male offsprings was remarkably incrcascd than the C group since 16 weeks (P ＜0.05) ; the blood glucose of LGTT (30 min) still had a marked rise until 24 weeks (P ＜ 0.05).(7) The blood pressure of male offsprings in G group had a marked rise on 12 weeks compared with the C group (P ＜ 0.05) ; from then on the blood pressure of G group kept up a rise trend until 26 weeks,but had no statistical significance (P ＞0.05).Conclusion The diabetic high-birth-weight rat model could be duplicated with STZ (25 mg/kg) once intrapertoneally on the first day of gestation,which were observed some evidently metabolic changes in weight,glucose tolerance and blood pressure.These results could represent an forward step in the clinical study of human gestational diabetes mellitus and their macrosomia babies,which may suffer some metabolic disease in their later life.

Objective:To explore the involvement of endoplasmic reticulum molecular chaperone GRP78 in the impairment of inner ear consistented with the mimetic aging model.Method:Twenty-four Wistar rats were randomly divided into two groups. model group was in duced by daily hypodermic injection of 10% D-galactose (800 mg·kg~(-1)·d~(-1)) for 8 weeks and the control group was given saline accordingly. Spatial learning and memory was measured by Morris-Water-Maze. Colorimetry was used to analyze superoxide dismutase (SOD) and malondialdehyde (MDA) extracted from inner ear tissue. Hearing threshold of rats were detected with Auditory brainstem response (ABR).In addition, expression of GRP78 in the inner ear was detected by immunohistochemistry,RT-PCR and Western blot. The control group was studied parallel.Result:The escape latency in the model group injected with D-galactose was markedly longer than that in the control group.accordingly ,the changes of SOD and MDA were more significant in the model group, the difference between two groups was significant(t-test,P<0.01). the variation of ABR in two groups was observed, There was no statistically difference of the hearing in the model group compared with the control group(P>0.05). The expression of GRP78 was significantly different between two groups ,which is increased in the inner ear tissue of model group(P<0.01).Conclusion:The impairment of inner ear tissue partly dued to the oxidative stress in the model, which was induced by D-galactose.and endoplasmic reticulum molecular chaperone was thought to contribute to the impairment mechanism of inner ear in mimetic aging model.

ObjectiveTo explore the effect of earlier rehabilitation on activities of daily living(ADL) of patients with spinal cord injury(SCI).Methods50 SCI patients received earlier rehabilitation and improvement of ADL of patients was evaluated.ResultsAfter two months treatment,the scores of Barthel index,functional independence measure(FIM) grade of patients increased significantly compared with that of before treatment(P<0.05) and ADL improved.ConclusionEarlier rehabilitation can improve ADL of SCI patients.

OBJECTIVETo investigate clinical outcomes of Kirschner wire as blocking screws combined with interlocking intramedullary nail internal fixation in treating tibial metaphyseal fractures (AO 43A).

METHODSFrom March 2011 to June 2012, 9 patients with tibial metaphyseal fractures were treated with blocking screws Kirschner wire combined with interlocking intramedullary nail, including 7 males and 2 females aged from 23 to 54 years old with an average of 37.4. Postoperative complications, X-ray were observed, AOFAS scoring were used to evaluate function after operation at 12 weeks.

RESULTSAll patients were followed up from 6 to 40 weeks (mean 20.1), and healed at stage I. No serious swelling, infection and skin necrosis occurred. No fracture instability and displacement appeaered at 4 and 8 week after operation. AOFAS score was (95.2±4.6) at 12 weeks after operation and 7 patients gained excellent result and 2 patients good.

CONCLUSIONKirschner wire as blocking screws with interlocking intramedullary nail for treatment of tibial metaphyseal fractures can fix well and perform simply.

Adult ; Bone Screws ; Bone Wires ; Female ; Fracture Fixation, Intramedullary ; methods ; Humans ; Male ; Middle Aged ; Tibial Fractures ; surgery

Bacterial Infections ; drug therapy ; Child ; Drug Resistance, Bacterial ; Humans ; Microbial Sensitivity Tests ; beta-Lactamases ; biosynthesis