Objective:To remove murine embryonic stem cells(mESC)from the differentiating cell culture and purify the differentiated cells by Magnetic Activated Cell Sorting(MACS).Methods:Neural differentiation of mESC was induced by a 5-stage method.The specific cell surface marker,SSEA-1,was used to identify ES cells in the differentiating cells.The optimal dilutions of mouse anti mouse SSEA-1 IgM primary antibody and FITC conjugated goat anti mouse secondary antibody were determined before the flow cytometry test.The incubation time and incubation temperature of primary antibody were all optimized to make the cytometry test accurate.After the optimization,stage 4 cells were dissociated into single cell suspension,incubated with antibody of SSEA-1 and microbeads conjugated goat anti mouse IgM,and then sorted through the magnetic field.The rate of SSEA-1 positive cells in pre-and post-separation groups was assessed by flow cytometry,and the viability of cells was evaluated by trypan blue staining counting under light microscopy.Results:The proportion of SSEA-1 positive cells in the separated cells can be reduced from(7.19?1.36)% to(1.34?0.80)%.The survival rate of sorted cells was more than 92%,similar to that of pre-separation cells.Conclusions:The MACS system we used can effectively remove mESC from the differentiated cells.The sorted cells will be well provided for the subsequent studies about transplantation therapy.

Objective To systemically review the effect of ulinastatin on lung function in pediatric patients undergoing cardiopulmonary bypass.Methods PubMed,Embase,Cochrane Controlled Trials Reg-istry,China National Knowledge infrastructure,China Biology Medicine disc,VIP and Wanfang databases were searched from their inception to October 2015.Articles regarding the use of ulinastatin on lung function in patients undergoing cardiopulmonary bypass were searched.Studies were screened by two independent re-viewers and then the data were extracted.The methodological quality was evaluated according to the inclusion and exclusion criteria.Meta-analysis was then performed using RevMan 5.1 software. Results Nineteen eligible studies (n = 657 patients)were identified.The results of meta analysis showed that ulinastatin could improve the oxygen partial pressure(SMD=0.90,95%CI 0.52-1.28,P <0.01)and oxygenation index (SMD=1.01,95%CI 0.45-1.56,P <0.01),decrease the PA-a O2 (SMD= -0.87, 95%CI -1.70--0.03,P =0.04),reduce the respiratory index (SMD=-0.81,95%CI -1.51--0.11, P =0.02),Lower the airway peak pressure (SMD=-0.83,95%CI -1.18--0.48,P <0.01),improve the dynamic compliance (Cd)(SMD=1.10,95%CI 0.57-1.62,P <0.01),and shorten the breathing ma-chine ventilation time (SMD=-0.98,95%CI -1.59--0.36,P <0.01).Conclusion This meta-analysis showed that ulinastatin treatment had a certain degree of protective effects on lung function in pediatric pa-tients undergoing cardiac surgery with CPB,but further research was needed for all these studies which were not multicenter,strictly controlled.

Objective To study the relationship between the intervals of Mitomycin C treatment and cytotoxicity, apoptosis and drug resistance for bladder cancer cells.Methods The bladder transitional cell cancer line BIU-87 was treated for two hours every time for five times with intervals of 24, 48, 72 and 96 hours respectively.Cytotoxicity was measured by MTT.p53,bcl-2,Bax and p170 expression were analyzed by Western blot.Results The IC_(50)(?g/ml)were 4.41,0.71,2.83,4.51and 6.16 with treatment intervals of 24, 48, 72 and 96 hours respectively, p53 and bcl-2 were significantly down-regulated and bcl-2/Bax was re- duced at 24 hour treatment interval but not changed at 48,72 and 96 hour intervals,p170 was not detected at 24 hour treatment interval but increasingly expressed at 48,72 and 96 hours intervals.Conclusion The in- terval of Mitomycin C treatment is closely related with cytotoxieity and apoptosis and drug resistance of blad- der cancer cells.The intervals of intravesical instillations may play an important role in the effect of chemotherapy.

Objective To assess the feasibility of low-dose MDCT pulmonary angiography (MDCTPA) by using low kVp combined with individual-adapted low tube current protocol and noise reduction filter algorithms.Methods Fifty-six consecutively patients with MDCTPA were enrolled in the study.The patients were randomly divided into group A (120 kVp,100 mAs) and group B (100 kVp,individual-adapted tube current about 1.0 mAs/kg).Two series of images were reconstructed in group B:B1 with standard filter algorithms (FC13,B1) and B2 with noise reduction filter algorithms (FC11,B2).The image noise,quality and radiation dose in two groups were compared.Results The CTDIvol,DLP and E of group B were 5.1 mGy,83.3 mGy· cm and 1.4 mSv respectively,which were reduced by 64.3％,66.4％ and 66.7％,respectively (F =32.57,12.32,11.98,P ＜ 0.05) when compared with those in group A.Compared with group A,the image noise in B1 was increased by 118.2％ (t =10.05 P ＜0.05),the central and peripheral SNR and CNR and scores were increased by 51.5％,46.6％,50.8％,45.3％and 20.4％,respectively(t =7.20,6.30,6.58,5.54,8.35,P ＜ 0.05).Compared with B1,the image noise in B2 was reduced by 32.5％ (t =6.12,P ＜0.05),the central and peripheral SNR and CNR and scores were increased by 46.2％,46.2％,45.9％,46.9％ and 18.4％ (t =3.19,3.55,2.95,3.37,5.42,P ＜0.05).Compared with group A,the image noise in B2 group was increased by 47.3％ (t =4.03,P ＜0.05),the central and peripheral SNR and CNR and scores were reduced by 29.1％,21.8％,28.2％,19.6％ and 8.2％(t =4.06,2.82,3.68,2.22,3.02,P＜0.05).Conclusions Keeping effective diagnostic image quality,the protocol with low kVp and individual-adapted low tube current for low-dose MDCT pulmonary angiography would be effective and feasible,and the radiation dose would be significantly reduced.

ObjectiveTo investigate the role of overexpressed proinflammatory cytokines in the pathogenesis of cerebral palsy (CP).MethodsLevels of tumor necrotic factor-α (TNF-α) and interleukin-6 (IL-6) in serum of 31 CP children, 20 healthy children (as controls), 37 neonates with CP risk factors such as hypoxic-ischemic injury and/or perinatal infection, and 20 healthy neonates (as controls) were measured by enzyme-linked immunosorbent assays (ELISA) retrospectively.ResultsLevels of TNF-α and IL-6 of CP children and neonates with CP risk factors were significantly higher than that of healthy controls (P<0.05). TNF-α level of CP children was significantly higher than that of neonates with CP risk factors (P<0.05), but there was no significant difference in IL-6 level between two groups.ConclusionOverexpressed proinflammatory cytokines play an important role in the pathogenesis of CP and may be an independent risk factor of CP.

OBJECTIVETo investigate whether PTEN can increase sensitivity of Ishikawa cells, an endometrial carcinoma cell line, to doxorubicin.

METHODSIshikawa cells transfected by PTEN gene or not were separately treated with serial concentrations of doxorubicin. The sensitivity of cells to doxorubicin was determined by MTT assay. The cells were stained with Hoechst 33258 and examined under fluorescence microscope to determine cell apoptosis. Immunoprecipitation and Western blotting analysis were performed to evaluate the effects of doxorubicin on phosphorylation of Bad and Akt/PKB.

RESULTSDoxorubicin induced cell death of the PTEN-transfected and non-transfected Ishikawa cells in a dose-dependent manner, but the cell death was more significant in PTEN-expressing clones than in parental Ishikawa cells. A low concentration of doxorubicin (0.1 micromol/L) did not affect cell apoptosis in PTEN-null Ishikawa cells, but it induced cell apoptosis in PTEN-expressing clones. A high concentration of doxorubicin (1 micromol/L) induced cell apoptosis in both cell lines. However, the percentage of apoptotic cells was higher in PTEN-expressing clones than that in parental Ishikawa cells. In the PTEN-expressing clones, expression of phospho-Akt/PKB and phospho-Bad (Ser-136) was down regulated. Doxorubicin reduced the levels of phospho-Akt/PKB and phospho-Bad (Ser-136) in both cell lines, but the most significant reduction occurred in the PTEN-expressing clones.

CONCLUSIONPTEN significantly enhances chemosensitivity of Ishikawa cells to doxorubicin. With PTEN expression, doxorubicin may exert apoptosis-induction activity by downregulation of the PI3k/Akt/PKB signaling pathway in Ishikawa cells.

Adenocarcinoma ; genetics ; pathology ; Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Endometrial Neoplasms ; genetics ; pathology ; Female ; Humans ; PTEN Phosphohydrolase ; biosynthesis ; genetics ; Transfection

OBJECTIVE:To investigate the effects of saikosaponin-d(SSd) on tissue plasminogen activator(TPA),plasminogen activator inhibitor(PAI),malonaldehyde(MDA) and NO in rats with liver fibrosis induced by dimethylnitrosamine(DMN).METHODS:Eighteen SD healthy rats were randomized to control group(NS ip qd for 4 weeks),model group(10mg? kg-1 DMN ip 3 times per week for 4 weeks) and SSd-treated group(10mg? kg-1 DMN ip 3 times per week + SSd 1.8 mg? kg-1 ip for 28 consecutive days).All rats were killed 1h after the last time of administration,blood samples were taken from abdominal aorta and liver samples were taken for the observation of pathology and detection of indices of TPA,PAI,MDA and NO etc.RESULTS:SSd could lessen the degree of liver fibrosis and improve the fibrinolytic activities of TPA and PAI,meanwhile,it showed clearance effect on MDA and marked protective effect on hepatic cells.There were significant differences between SSd-treated group and the model group.CONCLUSION:SSd exhibited protective function on experimental hepatic fibrosis in rats,which may be attributed to the improving of fibrinolytic activity,eliminating of lipid peroxidation and enhancing of NO level.

Objective To investigate the distribution and antimicrobial susceptibility of the bacteria isolated from bloodstream infections during 2013-2014 in Changhai Hospital for rational use of antibacterial agents.Methods The bacterial strains from blood samples were collected during the period from January 2013 through December 2014,and subjected to antimicrobial susceptibility testing by using automated system or Kirby-Bauer method.The results were interpreted according to CLSI M100-S24 breakpoints or FDA breakpoints.The data were analyzed by WHONET 5.6 software.Results A total of 1 048 nonduplicate isolates were collected,of which Escherichia coli,coagulase-negative Staphylococcus (CNS) and Klebsiella pneumoniae accounted for 29.5％,15.8％ and 13.8％,respectively.Gastroenterology,Hematology,General surgery,Urology and Department of Infectious Diseases are the top 5 departments according to their total number of bacterial isolates.The results of antimicrobial susceptibility testing showed that ESBLs-producing E.coli and K.pneumoniae accounted for 63.8％ and 38.6％,respectively.The prevalence of methicillinresistant CNS (MRCNS) was 77.6％.The E.coli strains isolated from Urology showed higher resistance rates to cephalosporins than the total E.coli strains,while the E.coli strains isolated from Gastroenterology showed higher resistance rates to betalactarn/beta-lactamase inhibitor combinations and carbapenems than the total E.coli strains.Higher prevalence of MRCNS was found in departments of Hematology,Urology and Neurosurgery.All the CNS strains isolated from Neurosurgery were resistant to methicillin.The K.pneumoniae strains isolated from Bum ICU had higher resistance rates to all the antibacterial agents tested than the total K.pneumoniae strains,while the K.pneumoniae strains isolated from Gastroenterology showed higher resistance rates to carbapenems and tigecycline than the total K.pneumoniae strains.Conclusions The pathogenic bacteria isolated from bloodstream infections vary with departments in terms of species distribution and antimicrobial susceptibility profile.It is necessary to strengthen the surveillance of antimicrobial resistance in hospital for rational use of antibiotics.

OBJECTIVETo observe the effects of Qufengtongluo Recipe (QFTLR) on the expression of podocin mRNA and podocyte morphology in rats with adriamycin-induced nephropathy (AN), and explore the possible mechanism mediating the therapeutic effect of QFTLR on nephropathic proteinuria.

METHODSSD rats were randomized into normal control group, AN model group (established by a single injection of adriamycin via the tail vein), and 3 intervention groups with QFTLR, prednisone, or benazepril treatment. After the corresponding treatments, the expression of podocin mRNA in the renal tissues was detected by RT-PCR methods, and the morphological changes of the podocytes were examined by electron microscope.

RESULTSCompared with the normal control group, the AN model group showed significantly lowered expressions of podocin mRNA (P<0.01) with reduced podocytes and widening, fusion or even absence of the foot processes (FP). Intervention with QFTLR significantly increased the expression of podocin mRNA (P<0.01) and the number of podocytes, and obviously lessened the structural changes of the FP.

CONCLUSIONQFTLR can produce therapeutic effect against nephropathic proteinuria possibly by up-regulating the expression of podocin mRNA and improving the morphological changes of the podocytes.

Animals ; Doxorubicin ; Drugs, Chinese Herbal ; pharmacology ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Male ; Membrane Proteins ; genetics ; metabolism ; Nephrosis ; chemically induced ; metabolism ; pathology ; Podocytes ; pathology ; Proteinuria ; etiology ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the efficacy and safety of Danlong Oral Liquid (DOL) combined Western medicine (WM) in treating mild-to-moderate bronchial asthma patients (heat wheezing syndrome) at acute onset.

METHODSTotally 480 mild-to-moderate bronchial asthma patients (heat wheezing syndrome) at acute onset were randomly assigned to two groups in the ratio 3:1, the treatment group (360 cases) and the control group (120 cases). All patients received basic WM treatment. Patients in the treatment group took DOL, 10 mL each time, 3 times per day for 7 days in total, while those in the control group took Kechuanning Oral Liquid (KOL) , 10 mL each time, 3 times per day for 7 days in total. Efficacy for asthma symptoms, lung functions and scores of TCM syndrome and/or main symptoms were evaluated.

RESULTSThe percentage of clinical control and significant effectiveness of asthma symptoms in the treatment group was significantly higher than that of the control group (77.36% vs 56.07%, P < 0.01). The percentage of clinical control and significant effectiveness of lung functions in the treatment group was significantly higher than that of the control group (74.28% vs 50.00%, P < 0.01). The anterior-posterior difference in scores of TCM syndrome was significantly superior in the treatment group than in the control group (-11.26 ± 4.70 vs -9.21 ± 5.09, P < 0.01). The anterior-posterior difference in scores of main symptoms was significantly better in the treatment group than in the control group (-6.58 ± 3.08 vs -5.16 ± 3.45, P < 0.01). The incidence of adverse reactions was significantly lower in the treatment group than in the control group [1.73% (6/346 cases) vs 10.17% (12/118 cases) , P < 0.05].

CONCLUSIONDOL combined WM was superior to KOL in treating mild-to-moderate bronchial asthma patients (heat wheezing syndrome) at acute onset.

Anti-Asthmatic Agents ; administration & dosage ; therapeutic use ; Asthma ; drug therapy ; Biomedical Research ; Drug Therapy, Combination ; methods ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Hot Temperature ; Humans ; Lung ; Medicine, Chinese Traditional ; Phytotherapy ; Respiratory Sounds ; Syndrome