Objective To investigate the efficacy,safety and localization of multilayer amniotic membrane transplantation(AMT) for corneal perforation associated with ulceration.Design A retrospective clinical case series.Participants Nine patients(9 eyes)with corneal perforation secondary to ulceration were enrolled into this study.These were little response to medicine,including bacterial keratitis(4 eyes ),fungal keratitis(2 eyes),fungal and bacterial mixed keratitis( 1 eye),virus keratitis( 1 eye),and Mooren's ulcer ( 1 eye).Size of perforation was 0.5～3.0 mm in diameter.Methods The AMT was applied to tamp the perforation,fill the ulcer and cover the surface of ulcer.After surgery,the medicine was continued to be used to treat the original corneal ulcer.The follow-up ranged from 6～20 months.In the suffering eyes,reformation of anterior chamber,healing of ulcer,complications and recurrence of ulceration were observed.Main Outcome Measures Of the postoperative eyes,reformation of anterior chamber,healing of ulcer,complications and recurrence of ulceration.Results The anterior chamber reformed at the first postoperative day in 9 eyes,and kept in normal depth in the follow-up time.At postoperative 2 months,the ulcer healed with sear and a smooth surface.Corneal thickness of the ulcer area recovered almost to normal.During follow up,no recurrence of ulceration or severe complications was noted.Conclusions Multilayer AMT is a safe and effective method for the management of small corneal perforation associated with ulceration,but the ulcer healed with scar.(Ophthalmol CHN,2008,17:101-103)

Objective To investigate the effeets of ulimstatin on expression of intestinal defemin-5 mRNAin the rat model of sepsis.Method The experiment was performed in pharmaco-laboratory of medical college,Sun Yat-Sen University.sixty Sprague-Dawley rals were randomly divided into control,sepsis,pretreated andtreated groups(n=15).Semis was induced in the mts of latter three groups by cecal lifo.and puncture(CLP).The rats of pretreated group received 25 000 U/kg ulinastatin 2 hours before operation and the rats of uli-nastatin treated groups received 50 000 U/kg ulinastatin 2 hours after operation.Some pieces of ileum mucosa weretaken 12 h after CLP.Tge pathological changes were observed and the expression of RD-5 mRNA was detectedwith RT-PCR.All data were managed by SPSS 13.0 software and arIaIyzed by using One-way ANOVA and LSD-ttest.Results The expression of RD-5 mRNA in the rats of sepsis group significantly decreased compared to col-trol(P<0.05).The expression of RD-5 mRNA of pretreated and treated groups sigificantly inereased comparedto sepsis group(P<0.05);pretreated groups had more increased expression of,RD-5 mRNA compared to treatedgroups(P<0.05).Conclusions The expression of intestinal RD-5 mRNA significantly decreases in sepsis,which could be improved by the treatment of ulinadtatin leading to intestinal mucosal protection of the siqnifleant.The pretreatment may be more effective than the theTapeatic treatment in the rat model of sepsis.

Aim: To compare the stability of Endostar~(TM) and endostatin under different temperatures and pH using polyacrylamide gel electrophoresis( PAGE) and Western blot and to compare the activity of Endostar kept at 4 ℃ and 37 ℃ by inhibition of endothelial cell proliferation. Methods: Endostar and endostatin expressed by Phicha pastoris were kept at 4 ℃, and 37 ℃ for 96 hours, respectively. The electrophoresis of the samples was detected by reduced and non-reduced PAGE. The results were further confirmed by Western blot with rabbit anti-Endostar polyclonal antibody. The inhibitory effect of Endostar stored at different temperatures on HUVEC was also exam-ined by cell-based assay. Results: Single band at 20 kD was detected in all lanes of SDS-PAGE gel loaded with endostatin and Endostar samples under reducing condition. In acidic native PAGE with three different pH values, endostatin showed a smear characteristic, whereas Endostar showed a unique band in acidic non-continuous native PAGE. Although the smear phenomenon was also observed under two conditions of constant native elec-trophoresis, the major band of Endostar could be detected. Similar electrophoretic behavior was found for endosta-tin and Endostar stored at both 37 ℃ and 4 ℃ . Western blot showed similar results to those by PAGE. Furthermore, Endostar stored at these two temperatures also had identical inhibitory effect on proliferation of HUVEC. Conclusion: Endostar and endostatin exhibit similar thermostability at regular conditions, but Endostar was more stable than endostatin expressed in P. pastoris under acidic condition.

Objective To investigate the incidence and pregnancy outcomes of premature rupture of membranes (PROM) in pregnant women in Beijing.Methods A retrospective multicenter study of 18 534 cases delivered in Beijing Obstetrics and Gynecology Hospital,Beijing Friendship Hospital,Daxing MCH Hospital and Tongzhou MCH Hospital from January 2011 to December 2011,was conducted.Results Among 18 534 cases,PROM occurred in 4 504 cases (24.30％),including 3 910 cases of in term PROM (21.10％) and 594 cases of preterm PROM (3.20％).The incidence of premature delivery was 6.17％ (1 144/18 534),and among 1 144 cases of premature delivery 547 cases were PROM (47.81％);the incidence of PROM was 22.75％ (3 957/17 390) in term delivery.The overall cesarean section (CS) rate was 48.50％ (8 989/18 534) and that in pregnant women with PROM was 35.55％ (1 601/4 504),but the CS rate in pregnant women without PROM was 52.66％ (7 388/14 030).The rate of postpartum hemorrhage was 13.12％ (210/1 601)in CS cases and 4.17％ (121/2 903) in vaginal delivery cases (x2 =121.361,P=0.000).The mean hospital stay for PROM was (5.3±2.9) d in CS cases and (4.3±2.3) d in vaginal delivery cases (t =-12.136,P =0.000).Conclusions Without severe maternal or fetal complications,the incidence of PROM is relatively high in Beijing region and PROM may not increase the maternal or fetal complications.Vaginal delivery is the main mode of delivery for PROM.Cesarean section may not cause less neonatal complications,but may lead to more postpartum hemorrhage and longer hospital stay.

Objective To explore the prognostic risk factors of low level malignant obstructive jaundice treated with transhapetic biliary drainage.Methods The clinical data of 142 patients with low level malignant obstructive jaundice received percutaneous transhapetic biliary drainage management from January 2010 to June 2013 were retrospectively analyzed.The study parameters included gender,age,tumor type,preoperative obstructive time,preoperative infection,drainage method,Child-Pugh grade,serum total bilirubin (TBIL),albumin (ALB),serum creatinine (SCr),the postoperative declining degree of bilirubin and postoperative antineoplastic therapy.The prognostic risk factors were evaluated.Results Single variable analysis showed that preoperative infection (P =0.006),Child-Pugh grade (P =0.004),SCr (P =0.043),the postoperative declining degree of TBIL (P =0.001) and postoperative antineoplastic therapy (P =0.015) were the related factors for survival time.The further Logistic regression analysis showed that preoperative infection (OR =3.729,95％ CI 1.332-6.363,P =0.040),Child-Pugh grade ≥ 10 scores (OR =0.513,95％ CI 0.375-1.276,P =0.018) and postoperative antineoplastic therapy (OR =0.668,95％ CI 0.210-2.026,P =0.038) were the related factors for survival time.Conclusion In treating of low level malignant obstructive jaundice with transhapetic biliary drainage,the preoperative infection,Child-Pugh grade and postoperative antineoplastic therapy may be the important related factors that affect the patient's survival time,to evaluate the prognosis of these patients has important reference meaning.

OBJECTIVE To investigate the positive rate of ?-lactamase,oprD2,aminoglycosides modifying enzyme and qacE△1 gene among clinical isolated strains of multi-resistant Pseudomonas aeruginosa in our hospital.METHODS P.aeruginosa was determined by VITEK,and MIC was determined by agar dilution method.TEM,IMP,VIM,oprD2,aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6″)-Ⅱ,ant(3″)-Ⅰ,ant(2″)Ⅰ and qacE△1 in strains were detected by polymerase chain reaction(PCR).RESULTS The positive rate of TEM,aac(3)Ⅱ,aac(6′)-Ⅱ,ant(3″)-Ⅰ and ant(2″)-Ⅰ in multi-resistant P.aeruginosa were 51.4%,48.6%,40.0%,54.3%,45.7% and 60.0%,respectively.No IMP and VIM genes were detected.CONCLUSIONS Positive rate of ?-lactamase, aminoglycosides modifying enzyme is high in 35 tested strains,and that should be paid more attention.

Objective To preparegraphene quantum dots (GQDs) and construct a novel biosensor for determination of dopamine (DA) and uric acid (UA).Methods The GQDs was prepared by carbonization method from citric acid as carbon sources.Differential pulse voltammetry was used by the modified electrodes to detect uric acid and dopamine,and the detection performance was evaluate.Collected three experimenters morning urine on July 29,2016.The proposed sensor was used for biological samples detection.Results The size of GQDs were between 3 to 5 nm,which was observed by transmission electron microscopy (TEM).The proposed sensor had good linear correlation of 50～600 μmol/L UA (r2 =0.996 6) and 2～ 240 μmol/L DA (r2 =0.992 5).In uric acid in urine samples' detection (n=3),RSD value was less than 3％.The standard addition recovery rates of UA and DA were in 95.7％～101.4％ and 96％～102％ respectively.Conclusion The biosensor based on GQDs for determination DA and UA had been constructed successfully.

BACKGROUND: Hundreds of mouse embryonic stem cell lines are established presently. Most of these cell lines are collected from 129, C57BL/6J and BALB/C mice. Success rate of creating embryonic stem cell lines with Kunming mice is very low. OBJECTIVE: To explore the method of isolation and culture of murine embryonic stem cells of kunming species to increase the efficiency of establishment of embryonic stem cell lines from Kunming species mouse. DESIGN, TIME AND SETTING: The cell experiment was performed at the Chongqing Key Laboratory of Neurology from April 2006 to June 2007. MATERIALS: Blastocysts of 3.5 days and 4 days from Kunming species mouse were collected. METHODS: The blastocysts of 3.5 days and 4 days from Kunming species mouse were cultured on the fibroblast cell feeder layers for 4-5 days, and the cells from inner cell mass were isolated and subsequently cultured in vitro in 2.5 g/L trypase-0.4 g/L ethylenediamine tetraacetic acid (EDTA) and 1 g/L trypase-0.2 g/L EDTA at room temperature for 2-4 minutes. Embryonic stem cell colony was mechanically straggled. MAIN OUTCOME MEASURES: Colony growth was observed and determined by alkaline phosphatase staining, OCT-4 staining and karyotype analysis. RESULTS: Adherence rate, inner cell mass frequency and cloning efficiency of 4 day post coitum (dpc) embryo were higher than those of 3.5 dpc embryo (P

Objective To explore the effect of embryonic stem cells(ESCs) differentiating into cardiomyocytes(CM) by cardiomyocytes.Methods The blastocytes of 3.5 d from Kunming mouse,cultured on mouse embryonic fibroblaste cell feeder layers for 4～5 d,the mouse ESCs were isolated and subsequently cultured.Day 2～3 embryoid bodies(EBs) were derived from ESCs of passage 3 to 5 and then incubated with neonatal cardiomyocyte(CM) to induce into cardiomyocytes and the expression of cardiac specific cardiac troponin-T(TnT) and ?-actin were detected by immunofluoresence.Results The rhythmic beating embryoid bodies were observed on day 3 and reached 93% of rhythmic beating embryoid bodies on 12 day in group 2.All the beating cardiomyocytes derived from ESCs expressed cardiac-specific proteins for TnT and ?-actin,and the group induced by direct cell contact acquired the highest differentiating ratio as 56.5%,and was the highest differentiating ratio compared with other groups(P

AIM: To investigate the effect of tetrandrine (Tet) on proliferation of human pterygium fibroblasts (HPF) in vitro and to search for a new method to prevent the recurrence after pterygium surgery.METHODS: With different concentrations (0 to 160μmol/L) of Tet acting on HPF cultured in vitro, the impact was observed at 24, 48, 72 hours respectively after Tet intervention. The MTT method was used to assay the biologic activities of Tet and inhibitive rate of cell growth. The expression of proliferating cell nuclear antigen (PCNA) in each group was detected by immunohistoche-mistry before and after Tet intervention.RESULTS:With different concentrations of 20, 40, 80 and 160μmol/L and acting for 24 to 72 hours, Tet could inhibit the proliferation of HPF in a dose- and time-dependent manner (P<0.05). After the intervention of Tet, the expression of PCNA protein declined. When the concentration of Tet was in the range of 20 to 160μmol/L, it was able to inhibit the expression of PCNA in a concentration-dependent manner (P<0.05).CONCLUSION:Tet could significantly inhibit the proliferation of pterygium fibroblasts, and the inhibitive action was in a dose- and time-dependent manner within a certain range of concentration. But in high concen-tration (>160μmol/L), Tet would have cytotoxity.