Objective To observe the intervention effect of Ginkgo biloba extract EGb761 on tert-butyl hydroperoxide ( t-BHP)-induced injury in human umbilical cord-derived mesenchymal stem cells ( hUC-MSCs) . Methods Proliferation of hUC-MSCs after primary culture was detected by methyl thiazolyl tetrazolium (MTT) assay, and then the optimal concentrations of t-BHP and EGb761 for oxidative stress injured MSC model were screened. Cell apoptosis was assessed by flow cytometry after Annexin V-fluorescein isothiocyanatel propidium iodide ( Annexin V-FITC/PI) staining. Content of malondialdehyde ( MDA) and superoxide dismutase (SOD) activity in hUC-MSCs were evaluated, and the expression levels of p53 and p21Cip1/Waf1 were analyzed by real-time fluorescence PCR. Results Pretreatment with 10~200 mg/L of EGb761 for 3 hours reduced the sensitivity of hUC-MSCs to t-BHP ( 100 μmol/L) induced proliferation inhibition, while EGb761 over 100 mg/L had no significant effect on enhancing the protection of hUC-MSCs . EGb761 at 100 mg/L prohibited hUC-MSCs apoptosis and MDA accumulation in hUC-MSCs induced by 100μmol/L of t-BHP acting for 6 hours, maintained the enzymatic activity of SOD, and decreased the expression of p53 and p21Cip1/Waf1 in hUC-MSCs with t-BHP-induced injury. Conclusion EGb761 is capable of protecting hUC-MSCs against oxidative stress injury, and its mechanism is probably related with the modulation of p53/p21 signal pathway.

OBJECTIVE To analyze the isolation rate and four consecutive years′(2004-2007) antibiotic-resistance trend of extended-spectrum ?-lactamases(ESBLs) producing Escherichia coli in urinary tract infection and to provide evidence for anti-infection therapy.METHODS The anti-microbial susceptibility test results of E.coli cultured from the midstream urine of outpatients and inpatients of our hospital from 2004 to 2007 were collected.Whonet 5.4 statistical analysis software was used for analysis.RESULTS During four years,no apparent change has been shown about antibiotic resistant rate of ESBLs producing E.coli.The isolation rate of this kind of E.coli increased year by year from 31% to 44% and showed high resistance to third generation cephalosporins and quinolones antibiotics,but was 100% sensitive to imipenem.CONCLUSIONS With the ESBLs-producing E.coli increasing year by year,the antibiotic-resistance rate of E.coli isolated from urinary tract infection is apparently increased and thus screening of antibiotics sensitive to enzyme-producing bacterial is the key point when the empirical therapy especially when the third and fourth generation cephalosporins are ineffective.

Background and purpose: E-cadherin (EC) is a type of glycoprotein, which could maintain epithelial polarity and cell-cell junction. It is thought to be associated with the development and progression of hepatocellular carcinoma (HCC). The study aimed to detect expression of uniting detection sEC and AFP before and after operation in patients, and to monitor recurrence of postoperative. Methods:The levels of sEC and AFP in 105 HCC patients were tested by enzyme linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) before and after operation as well as in 70 healthy controls. Theirs correlation with clinical and pathological factors was analyzed. We analyzed the level of plasma sEC and AFP of 47 patients who had been suffered from the hepatocellular carcinoma operation and relapse. Results: The levels of sEC and AFP in HCC patients before operation were significantly higher than in controls, which declined significantly one week after operation. The sEC level before operation showed significant correlation with TNM staging (P<0.01), edmondson grade, the carcerous thrombi within the portal vein and the capsule of carcinoma (P<0.05), but not with age of patients or the tumor size (P>0.05). While AFP only correlation with TNM staging (P<0.05), not with others factors (P>0.05). When the recurrence was confirmed, the level of sEC was much higher significantly than the level postoperation (P<0.01). Conclusion:The levels of sEC in HCC patients before operation are closely related with prognosis and recurrence of HCC. The unites detection of sEC+AFP may be a predictive factor for recurrence of postoperation.

Objective To explore the effect of salsalate on the glucose metabolism of obese mice induced with high fat diet (HFD). Methods Eight week-old male C57BL/6J mice were fed with HFD in combination with 0.5% of salsalate (SAL group, n＝5) or normal saline (control group, n＝5) for 40 days. The effect of salsalate on serum glucose level was examined by glucose tolerance test (GTT) and insulin tolerance test (ITT). The expressions of endoplasmic reticulumn related proteins, including CCAAT/enhancer-binding protein homologous protein (CHOP), endoplasmic reticulum-localized DnaJ 4 (ERDJ4), glucose regulated protein (GRP)78 and GRP94, were measured by qPCR and Western blotting. Results The random blood glucose level of obese mice were significantly lower in the SAL group than that in the control group (P0.05), and GTT showed that the mice in the SAL group had better glucose tolerance. However, there was no significant difference in fasting insulin level between the two groups. ITT showed there was no difference in the change of blood glucose after insulin stimulation between the two groups. The mRNA expressions of GRP78 and GRP94 and protein expressions of CHOP, ERDJ4, GRP78 and GRP94 were significantly lower in the SAL group than those in the control group (P0.05 or P0.01). Conclusion Salsalate can alleviate the hyperglycemia of obese mice induced with HFD by inhibiting endoplasmic reticulum stress, and the effect is independent of the insulin secretion.

Objective To assess the role of endoscopic ultrasound guided fine needle aspiration (EUS-FNA)for diagnosis of gastrointestinal tract lesions.Methods Sixty-eight patients underwent endoscopic uhrasonagraphy and EUS-FNA between May 2007 and Dec.2008.The result of cytology and/or pathology was compared with that of surgical finding and follow-up study.Results EUS-FNA was successfully performed on 62 patients with lesiorm in oesophagus(4 eases),stomach(19 case),rectum(19 cases),liver(3 case),mediastinum(4 cases)and lymph node(13 cases),and had a successful rate of 91.18%(62/68).Among them,the lesions in 40 patients(64.52%)were cytologically confirmed.The lesions in 10 out of 22 patients(35.48%)were pathologically confirmed.Compared with the results of pathology and long-term followe-up study,the sensitivity,specificity and positive predictive value of EUSFNA in diagnosis of lesions in gastrointestinal tract was 85.48%,100.00%and 90.91%,respectively,The positive predictive value and negative predictive value was100.00%and 57.14%,respectively.Conclusion EUS-FNA is a safe,effective and accurate method for diagnosis of lesions in gastrointestinal tract,and has an important role in cytological diagnosis.

Objective:To differentiate human embryonic stem cells ( hESCs ) into keratinocytes ( K-hESCs) and analyse the expression characteristics of biomarkers of K-hESCs.Methods: The hESCs of line H9 were seeded on matrigel in mTeSR1 medium.The hESCs were directly differentiated into kerati-nocytes in epithelial differentiation medium with bone morphogenetic protein 4, retinoic acid and N2 sup-plement.The karyotype of K-hESCs was analyzed, comparing the gene expression differences of K-hESCs with human gingival epithelial cells (HGECs), human immortalized oral epithelial cells (HIOECs) and HaCaT by Real-time PCR.Molecular characteristics of the cell differentiation were observed throughout the process by immunocytochemical techniques.Results:H9-hESCs were successfully differentiated into the cells that exhibited characteristics of keratinocytes in epithelial differentiation medium.The karyotype of K-hESCs was 46, XX; and the keratinocyte gene p63 expression in K-hESCs was significantly lower than that in HaCaT ( P<0.05) , but there was no significant difference of p63 expression in K-hESCs, comparing with that in HGECs and HIOECs ( P >0.05 ) .Conclusion: H9-hESCs could be directly differentiated into K-hESCs.The gene expression of K-hESCs was similar to that of epithelial cells in the early stage of monolayer cells differentiation with high proliferative activity.

The kinetics of nicotine degradation by O.intermedium DN2 and its application in tobacco waste were investigated. The results showed that the optimum temperature of nicotine degradation by O.intermedium was 30 ℃, the pH value was 6.5 and a mount of inoculum was 5 %. Under above conditions, the kinetics of nicotine degradation of initial concentration 500 mg/L was studied. The results indicated that the degradation process of nicotine with no-induced strain DN2 followed inverse S-shaped curve, and degradation process of nicotine with induced cells of DN2 followed Eckenfelder mode. The half life of nicotine degradation was 17.43 h and 4.10 h, respectively. And the results also showed that tolerance of O.intermedium DN2 to nicotine was up to 5000 mg/L when 0.1 % of glucose was added. Nicotine (2 220 mg/L) in extract of tobacco wastes degraded about 95.22 % by strain DN2 in 60 h incubation, indicating that strain DN2 was of application value in treatment nicotine pollution.

Objective To investigate the correlation between spatial memory and sleep architecture and hippocampal volumes in patients with chronic insomnia disorder.Methods Twenty-two chronic insomnia patients and 17 normal sleepers (controls) were selected to evaluate the subjective insomnia using Pittsburgh Sleep Quality Index (PSQI), Epworth Sleepiness Scale (ESS), Insomnia Severity Index (ISI) and the objective insomnia by polysomnography (PSG).The cognitive function was measured by Montreal Cognitive Assessment (MoCA).Spatial memory and object-memory were measured by Nine-box Maze, and object-recognition memory was detected by picture recognition test.MRI was used to detect hippocampus volumes.Results Compared with controls, a significant reduction in total sleep time (328.3 (310.4, 387.9) min vs 418.0 (375.8, 45.5) min, Z=2.607, P=0.009), sleep efficiency (%) (77.7 (73.1, 84.0) vs 93.0 (87.2, 93.9), Z=3.739,P=0.000), proportion of N3 (%) (5.5 (0.4, 14.4) vs 13.7 (7.7, 18.3), Z=2.664, P=0.008) and proportion of rapid eye movement (REM) (%) (14.4 (10.7, 17.2) vs 17.3 (15.9, 23.3), Z=2.890, P=0.004) was seen in insomnia patients, whereas sleep latency was delayed.The error numbers of spatial working-memory (4.5 (2.0, 7.3) vs 1.0 (0.0, 3.0), Z=3.007, P=0.003) in chronic insomnia patients were more than those in controls.There was no statistically significant difference in object reference memory, spatial reference memory and object recognition in two groups.A significant reduction of the left (2 818.0 (2 534.9, 3 191.8) mm3 vs 3 453.3 (3 081.2, 3 764.4) mm3, Z=3.314, P=0.001), right (2 780.5 (2 451.2, 3 191.8) mm3 vs 3 479.8 (3 024.1, 3 786.7) mm3, Z=3.484,P=0.000) and whole hippocampal volumes (5 561.7 (4 956.6, 6 396.9) mm3 vs 6 898.9 (6 017.1, 7 540.1) mm3, Z=3.455, P=0.001) was seen in chronic insomnia patients compared with controls.The hippocampal volumes were negatively correlated with sleep latency (r=-0.432, P=0.006), but positively correlated with sleep efficiency, proportion of N3 (r=0.323, 0.376;P=0.045, 0.018).There was a negative correlation between the error numbers of spatial working-memory and hippocampal volumes (r=-0.351, P=0.029).The hippocampal volumes were negatively correlated with the duration of disease in chronic insomnia patients (r=-0.734, P<0.01).Conclusion The spatial memory may be associated with decreased proportion of REM and reduced hippocampal volumes in chronic insomnia patients.

Objective To explore whether antenatal corticosteroids used can improve the maturation of choroid plexus capillaries in preterm mice.Methods This study was carried out in 2 groups of preterm mice.The study group was made of pregnant women receiving dexamethasone at a dose of 4 mg/(kg?time),1 time/d intraperitoneally.Study group was further subdivided into four subgroups according to the timing of the steroid administration:day 13,14,15 and 16 of pregnancy,each subgroup included 12 premature mice.All animals received a second injection 24 hours after the first one.The control group was given normal saline.The animals were operated to obtain premature mice.The choroid plexus ca-pillaries were assessed for integrity of their basement membranes by electronmicroscopy.Results In the study group,the maturity of the basement membrane of the choroid plexus capillaries was significantly better than that in control animals at any study time.The tendency to become intact.Thickness of the basement membrane.The protein granules to turn to more tightness.Conclusion These findings might provide an experimental basis for the use of antenatal steroids to decrease the incidence of periventricular hemorrhage in premature infants.

Objective To study the relationship between anti-transcription intermediary factor 1 family proteins (TIF1) autoantibodies profiles and cancer-associated dermatomyositis (CAM) and to define the diagnostic value of different subtypes of anti-TIF1 aotuantibodies for CAM.Methods The sera from 156 patients with dermatomyositis (DM),55 with polymyositis (PM),70 with systemic lupus erythematosus (SLE),60 with rheumatoid arthritis (RA),46 with primary Sj(o)gren syndrome (pSS),14 with systemic sclerosis (SSc),49 with kinds of malignancies and 40 healthy subjects were examined by immunoprecipitation assays followed by western blotting.Statistical analysis were performed using ANOVA,t test Mann-Wittney U and x2 test or Fisher exact test,nonparametric method was used to evaluate the sensitivity and specificity through calculating the area under the receiver operating characteristic curve (ROC).Results In summary,32 of 156 sera from patients with DM (20.5％) were positive for at least one anti-TIF1 autoantibodies.There are four subtypes of anti-TIF1 autoantibodies profiles existed in patients with DM,including 4 patients with only positive anti-TIF1-α (12.5％),20 with only positive anti-TIF1-γ (62.5％),7 with both positive anti-TIF1-α and anti-TIF1-γ (21.9％) and 1 with both positive anti-TIF1-β and anti-TIF1-γ (3.1％).However,only positive anti-TIF1-α (7.3％) was observed in 4 patients with PM.No patients with other CTDs as well as malignancy and healthy subjects were positive for these autoantibodies.The sensitivity and specificity of presence of anti-TIF1-α antibodies for the diagnosis of CAM were 42.9％ and 96.5％,respectively and those of anti-TIF1-β antibodies were 0 and 99.3％,respectively and those of anti-TIF-1-γ antibodies were 64.3％ and 86.6％,respectively.Application of areas of ROC to identify the best performance of test of anti-TIF1 antibodies profiles were 0.70,0.50,0.76,0.74 and 0.71,respectively.Conclusion Joint detection of antiTIF1 autoantibodies profiles can improve the diagnostic capbility for CAM.