[Objective]To establish a method to determine the contents of glucase and fructose in extracts of leaves of Lonicera hypoglauca Miq. using HPLC-ELSD, and apply this method to study the content change of glucose and fructose in different solvents extracts in leaves of Lonicera hypoglauca Miq. [Methods] The chromatographic separation was achieved on Lichrospher NH2 column using a mobile phase composed of a mixture of acetonitrile and water(90:10) at a flow rate of 0.8mL·min-1 and the column temperature was 25℃.The ELSD drift tube temperature was set at 70℃ and the carrying gas was N2 and the flow rate of the gas was 1.6L·min-1. [Results] Glucose and fructose from extracts of the leaves of Lonicera hypoglauca Miq. could be wel separated, and the linearity ranges of glucose and fructose were shown at the concentration of 0.5840~1.168mg·mL-1 and 0.8256~1.8576mg·mL-1, the average recovery rate of glucose and fructose were 96.88%(RSD=1.11%) and 97.55%(RSD=1.15%), the stability of glucose and fructose was 2.12%and 0.38%, and the repeatability was 1.31% and 0.24%. Among the water extract,50% alcohol extract and 95% alcohol extract,the content of the glucose and fructose in the water extract was highest,which was 2.19mg·mL and 4.10mg·mL-1, the 50% alcohol extract was second, which was 1.13mg·mL-1 and 2.12mg·mL-1, and the 95% alcohol extract was lowest, which was 0.45mg·mL-1 and 0.65mg·mL-1.[Conclusion] The method is simple, accurate and has good repeatability, and can be used to determine the contents of glucose and fructose in the extract of leaves of Lonicera hypoglauca Miq. In this batch of leaves of Lonicera hypoglauca Miq., the contents of fructose and glucose were of water extract>50%ethanol extract>95%ethanol extract.

Objective To evaluate the significance of suPAR,SCC-Ag in plasma and HPV16,18 in cervical secretion for monitoring pathogenetic condition and prognosis in patients with cervical cancer.Methods 206 cervical cancer patients blood and cervical secretion were collected.Plasma level of suPAR and SCC-Ag were measured by enzyme linked immunosorbent assay (ELISA) in health women and patients with cervical cancer.The expression of HPV16,18 of cervical secretion in control group and patients with cervical cancer were detected by fluorescence quantitative PCR.The correlations of the three indexes were analyzed.Results The plasma level of suPAR and SCC-Ag,the expression of HPV16,18 of cervical secretion in cervical cancer patients were obviously higher than those in health controls with statistical significance ((1.072 5±0.305 2) ng/ml vs (0.501 7±0.179 3) ng/ml,(0.980 6±0.162 7) μg/ml vs (0.261 4± 0.006 3) μg/ml and 53.89 ％ (90/167),46.15 ％ (18/39) vs 6.67 ％ (4/60),P ＜ 0.05).There was a positive correlation between plasma suPAR level and SCC-Ag level in invasive carcinoma of cervix patients (r =0.564,P ＜ 0.05).The plasma level of suPAR between in HPV16,18 positive group and in HPV16,18 negative group did not show difference (P ＞ 0.05).Conclusions In invasive carcinoma of cervix patients,there is a positive correlation between plasma suPAR level and SCC-Ag level.But it's not yet to conclude that plasma suPAR level of cervix invasive carcinoma patients is related to infection of HPV16,18.

The incidence of dysphagia after stroke is high.It may affect the rehabilitation of patients and increase mortality.The assessment is of great significance for early objective examination of dysphagia,in which the bedside screening tests are the most common assessment methods in the routine clinical work.This article reviews the advances in research on the commonly used bedside screening tests of dysphagia.

Objective To investigate the reliability and effect of using the chimeric flap retrieved by laparoscopic surgery to cover the large defect of the extremities.Methods The debridement and vacuum sealing drainage (VSD) were performed on 18 patients, who were admitted due to the defects of the extremities.The free chimeric peritoneal-deep inferior epigastric artery perforator (DIEP) flap assisted by laparoscopic surgery was transplanted to cover the defect with exposed tendons and/or skeleton.Results The remaining defects of the extremities were 12 cm × 8 cm-30 cm × 17 cm.The peritoneal component of the chimeric flaps measuring 8 cm × 6 cm -14 cm × 10 cm retrieved by laparoscopic surgery was used to cover the tendons, bones and joints.The deep inferior epigastric artery perforator (DIEP) flaps measuring 13 cm × 10 cm-32 cm × 18 cm allowed the cutaneous coverage of wounds.The chimeric flaps survived completely excepting two patients.The two patients experienced partial necrosis of the chimeric flaps and received skin grafting to achieve the wound closure.The function of the injured extremities recovered partially after 6-18 months of follow-up.No abdominal pain, distension, herniation,bulging and intestinal obstruction were recorded.Conclusion The chimeric flap assisted by laparoscopy is a helpful, safe and effective method for reconstruction of large wounds in extremities with exposed tendons and bones.

Objective To study the association between three single nucleotide polymorphisms (SNP; rs2241766,rs1501299 and rs12495941) variants of the adiponectin gene (ADIPOQ) and polycystic ovary syndrome (PCOS) in PCOS family trios.Methods A total of 224 unrelated PCOS probands,their biological parents were recruited.Anthropometric variables such as waist circumference (WC),hip circumference (HC),height and weight were measured in all subjects during the first visit to the outpatient department.Body mass index (BMI) and waist-hip ratio (WHR) were calculated.Serum fasting glucose (FBG),fasting insulin (FINS),total cholesterol (TC),triglycerides (TG),low-density lipoprotein and highdensity lipoprotein cholesterol (HDL) levels were measured.PCOS patients were divided into two groups based on BMh group A (BMI＜25 kg/m2) and group B (BMI≥25 kg/m2).Parents of PCOS patients were accordingly categorized into group C,D (fathers) and group E,F (mothers).The transmission disequilibrium test (TDT) was used to analyze the association between three SNP of ADIPOQ and PCOS.Results (1) A significant positive association was detected between SNP rs1501299 and PCOS (x2=7.093,P=0.008).However we failed to find significant overtransmission of the other two SNP rs2241766 and rs12495941 from parents to PCOS offsprings (x2=1.620,P=0.203; x2=0.713,P=0.398).(2) Linkage disequilibrium (LD) was analyzed in the subjects,rs1501299 and rs2241766 were in weak LD (r2=0.063,D'=0.621).(3) The levels of WC,HC,WHR,testosterone,TG,HDL and FINS were significantly differences between obese and lean PCOS patients (P＜0.05).While in fathers we only found WC,HC,TC levels being statistically different (P＜ 0.05).Mothers of obese PCOS patients had increased levels of FINS compared with mothers of lean PCOS patients (P＜0.05).The genotype frequencies of the three SNP were not different in obese and lean PCOS patients and their parents (P＞0.05).Conclusions TDT confirms that SNP rs1501299 in the ADIPOQ is significantly associated with the risk of PCOS in the Chinese Han population.The three SNP of the ADIPOQ were not associated with the obesity of PCOS.

Objective To investigate the effects of signal pathway inhibitors PD98059 and LY294002 on cell proliferation, apoptosis, expressions of phosphorylated extracellular signal-regulared kinase (p-ERK) and phosphorylated protein kinase B ( p-Akt) in endometrial carcinoma xenografts. Methods Human endometrial carcinoma Ishikawa cells were cultured in vitro. The effects of PD98059 and LY294002 on proliferation, apoptosis, and cell cycle distribution of endometrial cancer cells were detected by monotetrazolium ( MTT) assay and fluorescence-activated cell sorting technique. The models of xenografted tumor were established by the subcutaneous inoculation in 24 nude mice, and then they were randomly divided into 4 groups ( n = 6) , normal saline group, PD98059 group (PD group) , LY294002 group ( LY group) or PD98059 + LY294002 group ( PD + LY group) by intraperitoneal injections, respectively. The anti-tumor efficacy was evaluated by measuring tumor volume and tumor growth status. The histopathological change of tumor specimens was observed using HE staining and terminal deoxynucleotidyl transferasemediated dUTP-digoxigen in nick and labeling method (TUNEL) testing and the expression levels of p-ERK and p-Akt were detected by immunohistochemistry method. Results ( 1) The proliferation of Ishikawa cells were suppressed after treated by PD98059 and ( or) Y294002, in which A570 values of cells decreased showing both time-dependent and concentration-dependent manner ( LY294002: Fgroup = 9. 801, P = 0. 002; Ftime = 10. 398, P = 0. 001. PD98059: Fgroup= 8. 213, P = 0. 015; Ftime = 6. 839, P = 0. 036). Cell cycle distribution analysis revealed that percentage of Ishikawa cells at G0/G1 phase(Ftime =35.049, P= 0.004; Fgroup = 32. 024, P ＜0. 01) increased and percentage of S phase cells (Ftime = 7. 789, P = 0. 049; Fgroup = 30. 132, P ＜0. 01) decreased significantly. The percentage of apoptotic cells increased significantly among PD group, LY group and PD + LY group, in which there were significant difference [(63. 3 ±0.5)% vs (30. 7 ± 20. 1) % vs(40. 8 ± 1. 3) % ; F = 621. 059, P ＜ 0. 01]. (2) Compared with the control group, the increasing of transplanting tumor volume in the treated groups were obviously ( F = 23. 545 , P ＜ 0. 01) , and the inhibited rate of the tumor was higher in PD + LY group than that in PD group or LY group [(68 ± 9 ) % vs ( 32 ± 16 ) % or ( 38 ± 17 ) % ; F = 10. 283 , P ＜ 0. 05]. ( 3 ) HE staining shown that there were different degrees of necrosis for endometrial carcinoma cell in different groups. The apoptosis of tumor cells were significantly increased in treated groups by TUNEL testing [(13. 7 ± 1. 5)% , ( 14. 1 ± 1. 2)% , (29. 0 ± 1. 8 ) % ; F = 320. 344, P ＜ 0. 01]. Immunohistochemistry results demonstrated that the expressions of p-ERK and p-Akt in treated groups were lower than that in control group, of which LY + PD group was the lowest one. Conclusion The signal pathway inhibitors PD98059 and LY294002 could inhibit the growth of human endometrial carcinoma in vivo and in vitro, in which may induce cell apoptosis.

Olive oil is one of the important byproducts in agriculture. It is rich in oleuropein, which can be hydrolyzed into several bioactive phenolic compounds, including hydroxytyrosol (HT).There are many literatures have been confirmed that HT has significant pharmacological effects in the anti-inflammatory, anti-virus, regulation of metabolic disorders and treatment of degenerative diseases. However, HT has severe instability in vivo and in vitro. It is a challenge how to improve its stability through structural transformation or utilization of pharmaceutical means to processing. This paper will focus on biological activity of HT and its stability improvement, and provid some new ideas for expanding the research and promoting the development of HT in the field of Medicine.

Objective To comprae the difference of Sex hormone,serum and follicular fluid anti Mullerian hormone (AMH) level tube,number of retrieved egg,the number of cleavage rate,fertilization oocytes,the number of available embryos and total embryo on in vitro fertilization and embryo transfer (IVF-ET) of different reaction groups,to explore the relationship between AMH and pregnancy outcome of IVF-EH.Methods Eighty-one cases of IVF-ET assisted pregnancy were divided into low ovarian response group,normal ovarian response group and high ovarian response group according to the response of the super ovulation.Serum sex hormone and AMH of all patients were detected on the third day of menstruation,on the day of HCG injection,oocyte pick-up (OPU) and embryo transfer (ET),AMH in FF were detected too.The difference of oocytes,cleavage rate,embryo quality and the indicators mentioned above among the three groups were compared.All of the above indicators were compared between pregnancy group and unpregnancy group.Results (1) The level of AMH on the third day of menstruation ((1.76 ± 0.47) μg/L,(3.45 ± 1.01) μg/L,(6.34 ± 1.29) μg/L,F =2.435,P＜0.001),HCG injection ((1.09±0.13)μg/L,(1.29±0.29)μg/L,(2.64±0.51)μg/L,F=1.542,P=0.001) and embryo planting((1.32±0.33) μg/L,(1.62±0.39) μg/L,(1o 91±0.41) μg/L,F =1.573,P ＜0.001) all were statistically significant in three groups.On day of OPU,the levels of serum AMH ((0.95±0.21)μg/L,(1.15±0.29) μg/L,(1.74±0.41) μg/L,F =12.573,P＜0.001) and AMH in FF((5.82±1.19) μg/L,(6.92±1.05) μg/L,(7.79±1.39) μg/L,F =9.83,P＜0.001) were statistically differences in the three groups.(2)The number of oocytes were positively correlated with AFC,based AMH,levels of E2,P and AMH on the day of HCG injection and OPU,E2 and AMH on the day of ET and AMH in FF(P＜0.05).(3)AMH in FF in pregnancy group was (6.22±0.82)μ g/L,significantly higher than those without pregnancy group ((5.31 ±0.71)μg/L,P =0.037).Conclusions Levels of AMH in serum and follicular fluid are good predictors of ovarian response in IVF-ET.The level of serum AMH can only predict ovarian response,but can not effectively predict pregnancy outcome.The level of AMH in FF is a good predictor of pregnancy outcome.

Objective To investigate the influence of pertussis toxin(PTX)on G protein-coupled estrogen receptor(GPER)-mediated activation of phosphatidylinositol 3-kinase(PI3K)/protein kinase B (Akt)signaling activated by 17 β-estradiol(17β-E2)in endometrial carcinoma cells.Methods Expressions of GPER protein were detected by immunohistochemical SP method in Ishikawa and HEC-1A cells.Changes of levels of GPER,ERα and ERβ protein and the activation of Akt protein were observed by western blot in the two cells after they were treated by PTX for 30 minutes at different concentrations(0,0.1,0.5,1.0 μg/ml),and then co-stimulated with with 1 × 10-6 mol/L 17β-E2 respectively at different time (Ishikawa 30 minutes,HEC-1A 15 minutes).Results(1)Immunohistochemical SP method showed that GPER was positive stained in cell cytoplasm of Ishikawa and HEC-1A cell.(2)After co-treated with PTX at different concentrations(0,O.1,0.5,1.0 μg/ml)and 10-6 mol/L 17β-E2,in Ishikawa cell,the ratio of pAkt/Akt was 0.74 ±0.54,0.34 ±0.06,0.18 ±0.03,0.07 ±0.15,the gray values of GPER was 0.872 ± 0.490,0.395 ± 0.054,0.145 ± 0.014,0.034 ± 0.008,and with increasing concentration of PTX,the ratio of p-Akt/Akt and the expression of GPER decreased gradually(P ＜ 0.05),which was most obviously when the concentration was 1.0 μg/ml(F =63.729,P =0.0001;F =160.284,P =0.0001);ERα and ERβ protein had no significant change among different groups(P ＞0.05).In HEC-1A cell,the ratio of pAkt/Akt was 0.73 ±0.09,0.26 ±0.14,0.11 ±0.03,0,the Gray values of GPER is 0.927 ±0.134,0.485 ± 0.022,0.194 ± 0.004,0,and with increasing concentration of PTX,the ratio of p-Akt/Akt and the expression of GPER decreased gradually(P ＜ 0.05),which were also completely inhibited when the concentration was 1 μg/ml(F =1039.321,P =0.0001;F =109.646,P =0.0001),ERα protein had no significant differences(P ＞ 0.05)among different groups.ERβ was negatively expressed.Conclusion The results proposed that the activation of PI3K/Akt signaling in Ishikawa and HEC-1A cells could be inhibited after blocking the role of GPER by PTX.

A liquid chromatography_tandem mass spectrometric method was established for the determination of 25 kinds of veterinary antibiotics in manure, soil and water by ultrasonic_solid phase extraction. Purification efficiency of SAX cartridges for soil and manure samples extract and the procedure that manure samples defatted with hexane was also verified. The results showed that the average recoveries of the 25 target antibiotics were 50. 0%-121. 9%, the relative standard deviations (RSD) were 1. 1%-14. 71% (n=9), and the limits of detection (LOD) ranged from 0. 0002-0. 0560 μg/kg for soil and manure and ranged from 0. 002-0. 28 ng/L for water; After adding SAX cartridges, the matrix effects of manure samples reduced to 55%-120%, and soil samples to 75%-160%; after defatting by hexane extraction, the matrix effects of manure sample reduced to 55%-120%. This method has been employed to detect the veterinary antibiotics in environmental samples of livestock farm.