Objective To investigate pathological changes of lens epithelium cells (LECs) and expressions of matrix metalloproteinase-3 (MMP-3),alpha-smooth muscle actin (α-SMA),and fibronectin (Fn) in lens epithelial cells of experimental diabetic cataract rats,and to evaluate the roles of MMP-3,α-SMA,and Fn in the pathogenesis of diabetic cataract.Methods A total of 105 healthy Sprague-Dawley (SD) rats without lens diseases was randomly divided into normal control (n =45) and diabetic model (n =60) groups.Diabetic model rats were subjected to intraperitoneal injection of streptozotocin (STZ) (60 mg/kg),and those in the normal control group received injection with 0.1 mmol/L citric acid buffer solution of the same volume.The diabetic models were affirmed upon a fasting blood ≥ 16.65 mmol/L at the 3rd days after the injection.Once a week,the changes of blood glucose and body weight were monitored and the progression of cataract formation in both lenses of all rats was recorded with slit lamp observation.At end of 4 weeks,8 weeks,and 12 weeks after STZ injection,lenses were isolated and embedded in paraffin.The LECs histopathology was examined with HE staining.Immunohistochemical method was used to detect expressions of MMP-3,α-SMA,and Fn in normal and diabetic LECs.The related comparisons and statistic analysis were carried out.Results The lenses of control group were always completely transparent throughout the period of experiment with 31.48％,77.78％,and 100％ of lenses in diabetic model group swelling at 4th,8th,and 12th week,respectively.Under the light microscopic level,it has been showed that lens epithelium cells,which was occurred aggregate plaque and arranged in many layers,presented some morphologic characteristics of fibroblasts by HE staining.In control group LECs regularly,MMP-3,α-SMA,and Fn did not express equally;MMP-3,α-SMA,and Fn expressions increased obviously highly,difference had statistical significance compared to diabetic cataract group LECs (P ＜ 0.01).With the development of course of disease,the differences in expression of MMP-3,α-SMA,and Fn were significant between the diabetic cataract LECs and the control group (P ＜ 0.01).During the progression of diabetic cataract,the expression of Fn was positively correlated with that of α-SMA and MMP-3 (r =0.994,P ＜ 0.01;r =0.993,P ＜ 0.01).Conclusions The diabetic cataract liyingbody animal model was established successfully,which has laid down necessary basis to expound the morbidity mechanism of diabetic cataract.In the lens of diabetic cataract group,the expressions of α-SMA,MMP-3,and Fn were significantly increased.They participated in the occurrence and development of diabetic cataract.The expression of Fn was positively correlated with α-SMA and MMP-3.The lens epithelium cells which took on elongated aspect like fibroblast cells appeared epithelial-mesenchymal transformation (EMT),which mediated the occurrence and development of LECs fibrosis.

Objective To develop a satisfactory and reliable method for simultaneous determination of seven cephalosporin antibiotics (cefadroxil,cephalexin,cefoperazone,cefuroxime,cefradine,ceftazidime and cefotaxime) in water samples using solidphase extraction and high performance liquid chromatography.Methods The detection and reference wavelengths were 254 and 270 nm,respectively.The analytes were separated within 20 min by a gradient elution program.The optimized pretreatment of water samples was as following:using hydrophilic-lipophilic balance HLB as SPE cartridges,adjusting pH to 3 and adding 3.0 g NaCl per 500 ml water sample.Results The linear range of the method was 0.05-5.00 mg/L for ceftazidime,cefotaxime and cefuroxime,0.10-10.00 mg/L for cefadroxil,cephalexin and cefradine,and 0.20-20.00mg/L for cefoperazone.The coefficients for the analytes were above 0.99 and the limits of determination were 0.05-0.39 ?g/L.The recoveries of seven cephalosporin antibiotics in purified water were 86.64%-105.28%,and its relative standard deviation were 2.61%-11.64%.Conclusion The method was sensitive,accurate and repeatable,it is applicable to the determination of the cephalosporin antibiotics in waters.

Purpose To investigate the expression of Netrin-1 in ovarian serous carcinoma and its c1inicopatho1ogica1 significance. Methods Netrin-1 protein expression was assessed by immunohistochemica1 method in tissue specimens from 20 cases of benign ovari-an serous cystadenomas,13 cases of border1ine ovarian serous neop1asms and 32 cases of ovarian serous carcinomas( OSC). Results The positive proportion of Netrin-1 protein in OSC tissues was significant1y higher than those in border1ine and benign ovarian serous ne-op1asms(P<0. 01). The expression of Netrin-1 in OSC tissues was associated with tumor grade and c1inica1 stage(P<0. 05),but not associated with age,site,tumor size or 1ymph node metastasis( P>0. 05). Kap1an-Meier ana1ysis showed that the 5-year surviva1 rate of patients with Netrin-1 over-expression was significant1y 1ower than that of patients with 1ower expression( P<0. 05 ). Conclu-sions The high expression of Netrin-1 in OSC tissues indicates that Netrin-1 p1ays an important ro1e in cancer pathogenesis and deve1-opment and it may be a new assistant marker for prognosis of OSC.

Abstract Objective:To explore antitumor effect of NDV HN. Methods: pIRHN nucleic acid vaccin was constructed and was transfectedto Hela cells. Westem blot was used to analyssize the expression of pIRHN nucleic acid vaccine in eukarytic cell. The mode of cell death was de-tected by fluorescence microscope, gel electrophoresis and TUNEL assay. The effect of pIRHN nucleic acid vaccine on the contents of siaha acidin the Hela cell was examined. Results: pIRHN nucleic acid vaccine could be expressed in the eukarytic cell. pIRHN could induced apoptosisafter HeLa cells were transfected. The effect of antitumor responses of pIRHN was correlated with the contents of sialic acid in tumor cells. Con-clusion: The reults of our experiment will provide the theory evidence for the anti tumor mechamism of NDV.

Objective To investigate the efficacy of breviscapine in the treatment of vascular cognitive impairment.Methods A self-controlled trial was carried out in 36 patients with vascular cognitive impairment.36 cases were treated with breviscapine injection 50mg intravenous infusion daily for 3 weeks.At the same time,all cases were also given citicoline 0.75g/d and enteric-coated aspirin 100mg/d as routine treatment.The means of the evaluation on therapeutic effect included MMSE and ADL,and the adverse reactions were also observed.Results After the treatment,the MMSE scales increased from(18.75 ± 3.25)to(21.62 ± 3.58)(t=2.52,P＜0.05),Meanwhile,ADL scales declined from(45.65 ±3.36)to(42.33 ±4.18)(t=3.71,P＜0.05).There was significant difference on the MMSE and ADL between the pre and post therapy.No obvious side effects were found.Conclusion The results indicated that the breviscapine injection is an effective medicine in the treatment of vascular cognitive impairment,while the adverse reactions were few.

Objective To evaluate the accuracy of point-of-care testing (POCT) for blood glucose monitoring in critically ill patients.Methods Two hundred and forty critically ill patients,of both sexes,aged 20-88 yr,with Acute Physiology and Chronic Health Evaluation Ⅱ score of 1-45,were enrolled.The venous,arterial and capillary blood samples were collected to determine the real-time blood glucose level using glucose oxidase (GOD) and glucose dehydrogenase (GDH) methods.The blood glucose level measured by central laboratory hexokinase method simultaneously was served as standard level.Error Grid analysis (EGA) and Bland-Altman analysis were used to determine accuracy and consistency,respectively.The accuracy of real-time blood glucose levels within the consistent limits was evaluated.Results 1.The results of EGA showed that 98.7 ％,98.3 ％,98.3 ％(GDH method) and 96.2％,96.6％,96.7％ (GOD method) of the difference between venous,arterial and capillary blood glucose levels measured and the standard level were located in the A and B zones,respectively,and 1.2％,1.7％,1.7％ (GDH method) and 2.9％,3.3％,3.3％ (GOD method) in the D zone.0.8％ (GOD method) of the difference between venous blood glucose levels and the standard level were located in the C zone.2.Bland-Altman analysis showed that the difference between the standard level and glucose level measured in blood samples from the vein,artery and capillary.was-0.1,-0.3,-0.2 mmol/L (GDH method) and-0.9,-1.0,-0.9 mmol/L (GOD method),respectively,and the incidence beyond the upper and lower limits of consistency zone was 4.5 ％,6.7 ％,6.6 ％ (GDH method) and 4.6 ％,5.0 ％,7.1％ (GOD method),respectively.The accuracy of venous,arterial and capillary blood glucose levels within the consistent limits was 94.3 ％,92.1％,93.7％ (GOD method) and 96.6％,95.1％,95.5％ (GDH method),respetively.Conclusion The accuracy of POCT for blood glucose monitored by GOD and GDH methods is good in critically ill patients,but it is possible to overestimate the patient's real glucose level.

Objective To investigate the effect of ultrasound combined with intravenous terbutaline sulfate for external cephalic version and the impact of this therapy on maternal and neonatal complications.Methods 126 singleton pregnant women who were diagnosed by B-ultrasound breech pregnancy at 34-36 weeks in regular antenatal examination from January 2014 to June 2016 were randomly divided into an observation group (n =70)and a control group (n =50).The observation group was treated with intravenous drip of terbutaline sulfate after epidural anesthesia,and then received external cephalic version under B-ultrasound monitoring.The control group received traditional knee-chest position for correcting breech presentation.The success rate of breech correction,breech version rate,cesarean section rate,vaginal delivery rate,adverse reactions,postpartum hemorrhage volume,neonatal asphyxia,and neonatal birth injury were noted and compared between the two groups.Results As compared with the control group,the observation group had a higher success rate of breech correction and a higher rate of vaginal delivery while a lower rate of cesarean section,with a significant difference (x2 =20.210,8.564;P ＜0.05).The rate of postpartum hemorrhage,neonatal asphyxia,or neonatal birth injury did not differ significantly between the two groups (P ＞ 0.05).Conclusions Application of ultrasound combined with intravenous terbutaline sulfate in external cephalic version is easy to operate and has a higher success rate and greater safety,significantly reducing the rate of cesarean section and improve the success rate of vaginal delivery.

This study was aimed to investigate effects of Er-Xian (EX) decoction in follicular granulosa cell apoptosis in chemotherapy-induced premature ovarian failure (POF) of rats. SD rats were randomly divided into the normal group, model group, positive group and EX decoction group. Intraperitoneal injection of cisplatin was used in the establishment of chemotherapy-induced POF rat model. Intragastric administration of corresponding medication was give to each group for four weeks. The general conditions of rats were observed. The serum contents of E2, FSH, LH and Pro were determined. The ovarian tissue morphology was observed. Granulosa cell apoptosis was analyzed by TUNEL. The expressions of BAX, Bcl-2 and VEGF protein were detected by immunohistochemistry. The results showed that in the model group, the body weight decreased with disordered estrous cycle. After treatment, the weight gained, and the estrous cycle recovered. Compared with the normal group, in the model group, E2 decreased, FSH and LH levels increased. There was no significant difference in the content of progesterone among different groups. Detection of TUNEL showed that the positive expression in the model group was increased compared with the normal group (P < 0.05). The positive expression of EX decoction group was reduced compared to the positive group (P < 0.05). Compared with the normal group, the expression of BAX was obviously increased and Bcl-2 was decreased in the model group. After EX decoction treatment, the expression of BAX was decreased and the Bcl-2 was increased compared to the model group. The expression of VEGF protein in the model group was obviously less than that in the normal group. It was concluded that cisplatin can induce follicular granulosa cell apoptosis, and cause premature ovarian function recession. EX decoction can adjust the content of different hormones in serum, alter expression of apoptosis related protein to reduce the damage of cisplatin on ovarian follicular development, in order to promote follicular development, improve and enhance the ovarian function.

Objective:To investigate whether miR-9 plays a role in regulating glycogen synthase kinase-3β (GSK-3β) expression, affecting Wnt/β-catenin pathway activity and the patho-genesis of Osteo-arthritis (OA).Methods:The cartilage tissue of OA patients and normal cartilage tissue after traumatic amputation were collected, and the expressions of miR-9 and GSK-3β were compared. The double luciferase gene reporting test verified whether there was a targeted regulatory relationship between miR-9 and GSK-3β. OA rat model was established and compared with sham group, enzyme-linked immuno sorbent assay (ELISA) was used to detect hydroxyproline (Hyp) content in joint fluid.A kit was used to detect caspase-3 activity, and miR-9 and GSK-3β expression differences were detected in cartilage tissue. The OA model rats were divided into 3 groups: the sham group, the OA+ antagomiR-NC group, the OA + antagomiR-9 group. ELISA was used to detect Hyp content in joint fluid, kit was used to detect caspase-3 activity, and flow cytometry was used to detect cell cartilage tissue. Apoptosis, quantitative real-time polymerase chain reaction (qRT-PCR) and western blot were used to detect the expression of miR-9, GSK-3β, β-catenin and COL2A1. The comparison of mea-surement data between the two groups was conducted by t-test. The comparison of measurement data between multiple groups was conducted by one-way Analysis of Variance (ANOVA) analysis of variance, and then Bon-ferroni method was used for comparison between the two groups. P<0.05 was considered as statistically sign-ificant. Results:The miR-9 expression of cartilage tissue were (1.09±0.25) in the control group, and (2.86±0.25) in the OA group ( t=24.30, P<0.01). The GSK-3 β mRNA expression of cartilage tissue was (0.99±0.11) in the control group, and (0.53±0.10) in the OA group ( t=15.40, P<0.01). There was a targeted regulatory relationship between miR-9 and GSK-3β. The miR-9 expression of cartilage tissue was (1.00±0.21) in the sham group, and (2.61±0.36) in the OA group (t=9.462, P<0.01). The GSK-3 β mRNA expression of cartilage tissue was (1.00±0.18) in the sham group, and (0.52±0.09) in the OA group ( t=5.842, P <0.01). The Hyp content of joint fluid was (10±3) ng/ml in the sham group, and (50±8) ng/ml in the OA group ( t=11.015, P<0.01). The Caspase-3 activity of cartilage tissue was (1.00±0.19) in the sham group, and (2.43±0.36) in the OA group ( t=8.605, P<0.01). The miR-9 expression of cartilage tissue was (2.86±0.31) in the OA+antagomir NC group, and (1.67±0.19) in the OA + antagomir-9 group ( F=105.2, P<0.01). The GSK-3β mRNA expression of cartilage tissue was (0.41±0.09) in the OA antagomir NC group, and (0.81±0.09) in the OA + antagomir-9 group ( F=49.32, P<0.01). The Hyp content of joint fluid was (52.3±6.8) ng/ml in the OA + antagomir NC group, and (30.3±3.4) ng/ml in the OA + antagomir-9 group ( F=119.7, P<0.01). The caspase-3 activity of cartilage tissue was (2.22±0.23) in the OA + antagomir NC group, and (1.43±0.14) in the OA+ antagomir NC group ( F=72.55, P<0.01). Compared with OA + antagomir NC group, the expression of β-Catenin protein in the OA + miran-tagomir-9 group wasdecreased, the expression of GSK-3 β and COL2A1 protein wasincreased, and cell apo-ptosis wasdecreased. Conclusion:The increased expression of miR-9 plays a role in reducing the expression of GSK-3β, enhancing the activity of Wnt/β-catenin pathway, promoting the degradation, destruction of cartilage matrix and the pathogenesis of OA. Inhibition of miR-9 expression can reduce the protective effect of OA.

Objective To investigate the effect of penehyclidine hydrochloride on endotoxemiainduced intestinal injury in neonatal rats.Methods Ninety healthy neonatal Sprague-Dawley rats,aged 7days,weighing 16-20 g,were randomly divided into 3 groups (n =30 each) using a random number table:control group (group C),lipopolysaccharide (LPS) group,and penehyclidine hydrochloride group (group P).Endotoxemia was induced by intraperitoneal LPS 5 mg/kg in LPS and P groups.In group P,penehyclidine 2 mg/kg was injected intraperitoneally at 30 min before and after LPS injection.The equal volume of normal saline was given in group C.At 2,6 and 12 h after LPS or normal saline administration,10 rats in each group were randomly chosen and sacrificed.The ileum was removed to detect wet/dry lung weight ratio (W/D ratio) and contents of tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6),hypoxia-inducible factor 1 alpha (HIF-1α) and glutamine (Gln),and for examination of the pathologic changes of the ileum with light microscope.Results Compared with group C,the W/D ratio and contents of TNF-α,IL-6 and HIF-1α were significantly increased,and the content of Gln was decreased at each time point in LPS and P groups.Compared with group LPS,the W/D ratio and contents of TNF-α,IL-6 and HIF-1α were significantly decreased,the content of Gin was increased at each time point,and the pathologic changes of the ileum were mitigated in group P.Conclusion Penehyclidine hydrochloride can reduce endotoxemia-induced intestinal injury in the neonatal rats,and down-regulated expression of HIF-1α,up-regulated expression of Gln and attenuated inflammatory responses may be involved in the mechanism.