Objective To investigate the molecular epidemiology of 114C. albicans strains isolated from the vaginal discharge of female patients treated in three obstetrics and gynecology hospitals in Shanghai by analyzing the relationship between the main genotypes and resistance proifle, and the relationship between genetic diversity and cluster ofC. albicans.Methods A total of 114 strains ofC. albicans were collected from the Obstetrics & Gynecology Hospital of Fudan University, Shanghai First Maternity and Infant Hospital Corporation and the International Peace Maternity & Child Health Hospital of China welfare institute. Phylogenetic analysis of strains were carried out by eBURST.C. albicans strains were also analyzed by multilocus sequence typing (MLST). The susceptibility of theC. albicans strains was tested by ATB FUNGUS 3.Results A total of 47 diploid strain types (DSTs) were identiifed from the 114 strains, 30 of which were known types. DST 79 and DST 435 were the main types. Of the 114C. albicans strains, 96.5% were susceptible to lfucytosine, 100% to amphotericin B, 85.1% to lfuconazole, 55.2% to itraconazole and 84.3% to voriconazole.Conclusions The pathogenicC. albicans strains isolated from different obstetrics and gynecology hospitals in Shanghai were originated from multiple clones, the main type of which was DST 79 and DST 435 with certain degree of antifungal resistance. MLST typing suggests that genetic diversity is present in theC. albicans strains isolated in Shanghai area. The clustering analysis ofC. albicans strains is consistent with its genotypes.

Objective To establish a method for quality control of oleanolic acid and ursolic acid in Qingdu tablet. Methods The content of oleanolie acid and ursolic acid in Qingdu tablet were determined by high performance liquid chroma-tography(HPLC). The analytical column was kromasil-C18 (250 mm × 4.6 mm, 5 μm). The mobile phase consisted of methanol-0.5% phosphoric acid solution (88:12). The UV detection wavelenghth was 210 nm. The flow rate was 1.0 ml/min. Results Oleanolic acid and ursolic acid showed a good linearity in range of 0.06-0.54μg (R=0.999 3) and 0.30-2.70μg (R=0.999 7). The average recovery rates of Oleanolic acid and ursolic acid were 100.48% and 102.21%, respectively. Conclusions This method is simple, rapid and accurate. It could be used for the quality control and quality study of the Qingdu tablet.

Objective Study on methylenetetrahydrofolate reductase(MTHFR)gene polymorphism in coronary heart disease.Methods Detect the MTHFR C677T gene polymorphism by PCR-RFLP in 73 cases of healthy individuals and 87 cases of coronary heart diseas(CHD).Results At the site of 677,the T allele frequencies are 18.5%,36.1%,respectively,for healthy individuals and 87 cases of coronary heart disease.The frequecies of MTHFR CT genotype and T allel were significantly higher in disease groups.There are significant differences between disease group and control group(P

Objective To investigate the changes in cells on cell cycle,cell apoptosis and susceptibility to chemotherapeutic drugs of lung adenocarcinoma A549 cells under two dimensional (2D system) and three dimensional (3D system) culture conditions. Methods The three dimensional culture of A549 cells was carried out using rotatable culture bottle and desk-top water bath shake. Flow cytometer,in situ cell apoptosis detection kit,MTT and blood cell counter were employed to compare cell cycle,cell apoptosis and susceptibility to ADM of A549 cells under different culture conditions. Results The three dimensional culture system was confirmed to have been successful by the aid of observation under inverted and electron microscope. There were significant retardation of G_ 1 phase,lower cell apoptosis rate and decreased susceptibility to ADM in A549 cells in 3D system compared with those in 2D system. Conclusions There were remarkable differences in biological characteristics of A549 cells in two culture systems,indicating that A549 cells cultivated in 3D system simulated better solid tumor in vivo . The 3D system was very useful for further investigation of the behavior of solid tumor,so that anti-carcinoma chemotherapeutic drugs could be advantageously tested in vitro before clinical application.

Objective To investigate the protective effect of c-jun N-terminal kinase (JNK)inhibitor SP600125 against acute liver failure in mice.Methods Fifty-five male C57/BL6 mice were divided into control group (n =30) and SP600125 group (n =25).The animals were given an intraperitoneal injection of D-galactosamine (D-GalN,400 mg/kg body weight)/lipopolysaccharide (LPS,30 μg/kg body weight).The control group and SP600125 group were given 10％ dimethyl sulfoxide (15 mL/kg body weight) or SP600125 (75 mg/kg body weight) subcutaneously 12 h and 1 h before D-GalN/LPS administration,respectively.D GalN/LPS induced mouse JNK activation was detected by immunohistochemistry for phospho JNK (p-JNK).D-GalN/LPS induced mouse liver cell apoptosis was detected by immunohistochemistry for Caspase-3 and TdT-mediated-dUTP nick endlabeling (TUNEL).Serum alanine transaminase (ALT) level was tested to assess liver injury.Survival rate of mice within 24 h after D-GalN/LPS administration was observed.The comparison between groups was done by t test and survival rate was analyzed by Kaplan-Meier method.Results JNK activity in liver tissues,as indicated by observation of p-JNK positive cells by immunohistochemistry,was diminished 4 h after D-GalN/LPS administration in SP600125 group.Reduced Caspase-3 activity was observed 6 h after D-GalN/LPS administration in SP600125 group (as indicated in immunohistochemistry by Caspase-3 positive cells).Mice in SP600125 group showed significantly lower TUNEL-positive cell count than control group (43.0±24.5 vs 194.7±73.8; t=9.743,P=0.000).Serum ALT level 6 h after D-GalN/LPS administration was (24.0±54.7) U/L in SP600125 group,which was significantly lower than that in control group [(1234.4±478.4) U/L; t=4.734,P=0.0015].SP600125 also significantly improved the survival rate within 24 h after D-GalN/LPS administration (4/5 vs 1/10； x2=5.225,P=0.0223).Conclusions JNK inhibitor SP600125 exerts protective effects against D-GalN/LPS induced acute liver failure in mice by suppressing JNK activation and hepatocyte apoptosis.

Objective To explore an optimal model of three dimensional in vitro cell culture for simulating solid tumors in vivo . Methods The model of three dimensional cell culture was constructed under the conditions of inhibiting the cell wall attachment and stirring the medium. Multicellular spheroids (MCS) were cultured using microcarriers (CutiSpher). Drug sensitivity of monolayer cells (MC) and MCS was tested by MTT staining and cytometry, respectively. Ultrastructures of the MC and MCS were observed by transmission electron microscopy. Results Cells in three dimensional cell culture model without microcarriers were compacted into mass at 4 d while cells in MCS were found to attach to the microcarriers at 0.5 h. MCS had more than two layers of cells growing within it at 5 d. Compared with MC, MCS was more resistant to the anticancer drug, and had more plenty of organell and microvilli with more extensive and compact cell adhesion. Conclusion MCS has strong developmental properties and can simulate the cell cell interactions in vivo , especially cell adhesion, which may contribute to the drug resistance of MCS.

Objective:To explore the clinical effect of mild hypothermia combined with puncture aspiration operation on the hypertensive intracerebral hemorrhage and its effect on the serum TNF-α,IL-6 and CRP levels.Methods:112 cases of patients with hypertensive intracerebral hemorrhage admitted in our hospital from February 2015 to August 2016 were selected as the research objectives and randomly divides into 2 groups with 56 cases in each group.Both groups were treated with minimally puncture aspiration,and mild hypothermia was performed in the observation group additionally,the clinical effect,complication,short-term prognosis,serum TNF-α,IL-6,CRP levels at different time points before and after operation were compared between two groups.Results:The overall effective rate of observation group was 92.86％,which was significantly higher than of the control group(76.79％.P＜0.05).The incidence of MODS and complications were 0％ and 26.79％ respectively,which were significantly lower than those of the control group(P＜0.05).The serum TNF-α,IL-6,CRP levels of observation group were significantly decreased on the 3rd and 7th day postoperation,which were significantly lower than those before operation (P＜0.01).The serum TNF-α,IL-6,CRP levels of control group on the 3rd day postoperation were similar to those before operation (P＞0.05),but the serum TNF-α,IL-6,CRP levels on the 7th day postoperation were significantly decreased (P＜0.01).The serum TNF-α,IL-6,CRP levels on the 3rd,7th day postoperation were significantly lower than those of the control group at same time points (P＜0.01).The favorable prognosis of observation group was 80.36％,which was significantly higher than that of the control group (62.50％,P＜0.05).Conclusion:Mild hypothermia combined with puncture aspiration operation could effectively reduce the neurological deficits,inhibit the inflammatory response and decrease the mortality.

Objective To find an effective method for the treatment of mitomycin C extravasation injuries. Methods The rabbit model of mitomycin C extravasation was made and managed by different methods. The therapeutic efficacy in each group was assessed by using regression index and regression time. Results The therapeutic efficacy in ice compress group and ice compress plus amifostine group was better than that in other groups and the regression time was the shortest. There was no inflammation and necrosis in skin in ice compress group. Conclusion Ice compress, which can prevent inflammation and necrosis in skin, is one of the most effective treatment methods for mitomycin C extravasation injuries.

Objective To study the mechanism of multicellular drug resistance of pulmonary adenocarcinoma cell line A549 mediated by cell-cell adhesion. Methods We compared the sensitivity of monolayer cells (MCs) to adriamycin (ADM) with that of multicellular spheroids (MCSs) which was employed as a three dimensional cell culture model. Transmission electron microscopy was applied to observe the cellular ultrastructure. Bcl-2, Bcl-xL, and Bax expression levels were detected by flow cytometry and indirect immunofluorescent staining. Results Compared with MCs, MCSs had more than two layers of cells, more extensive and compact cell adhesion, and inlay junctions were found within them. MCSs were more resistant to ADM. At the same time, Bcl-2 and Bcl-xL expressions in MCSs were much higher. After treatment with ADM, expressions Bcl-2 and Bcl-xL increased markedly in MCSs. Conclusion MCSs could simulate the solid tumors in vivo and has multicellular drug resistance mediated by cell-cell adhesion. The possible mechanisms may be associated with the upregulation of Bcl-2 and Bcl-xL.

Objecive To explore the medicine packaging improvement based on PAP shelter hospital requirements to fulfill emergency medicine support,Methods The medicine support during rescue was discussed from the aspects of considerations in medicine plan,requirement for emergency medicine,special medicine purchasing and medicine support,and some countermeasures were put forward for improving medicine packaging.Results Its suggested that the optimization of medicine packaging be performed with considerations on medicine property,package capacity,convenience,transport and environmental suitability.Conclusion Emergency medicine support is of importance for disaster medical rescue,and medicine packaging improvement based on PAP shelter hospital requirements contributes to enhancing the efficiency during disaster rescue.