Objective To investigate the inhibitory effect of methionine-γ-lyase(METase) gene MEGL on glioblastoma(GBM) cells and its potential molecular mechanism,and to provide new ideas for the development of MEGL drugs.Methods MEGL expression system mediated by lentivirus was constructed and used to infect human GBM cell lines U87 and snb19,and the cells infected with empty virus were as the control.RT-qPCR and Western blot were used to detect the expression of MEGL gene and protein.The levels of methionine(Met) and its metabolites were detected by liquid chromatography tandem mass spectrometry(LC-MS/MS).The cell proliferation was measured by cell counting method.The changes of cell cycle were detected by flow cytometry,and the expression of cell cycle-related proteins(CDK2,CDK4 and p21) and mitogen-activated protein kinase(MAPK) signaling pathway proteins was determined by Western blot.Results Met levels in U87 and snb19cells stably expressing MEGL gene decreased significantly(t=10.996 and 15.733,respectively,each P <0.001).Furthermore,MEGL significantly suppressed the proliferation of U87(on the 3rd,4th and 5th day,t=14.698,9.746,and 12.263,respectively,each P<0.001) and snb19 cells(on the 3rd,4th and 5th day,t=8.480,6.939,and 13.258,respectively,each P <0.001),inducing a notable increase in the proportion of G0/G1 phase(t=7.997,P P<0.001).MGEL gene decreased the expression of cell cycle related proteins CDK2 and CDK4(t=8.471 and 3.369,P=0.0011 and 0.0281,respectively),while promoting the expression of p21(t=10.761,P <0.001).Moreover,it significantly inhibited the expression of proteins related to the MAPK signaling pathway,including p-p38,p-ERK1/2,and p-JNK(t=3.401,6.493,and9.413,P=0.027 2,0.002 9,and <0.001,respectively).Conclusion The MEGL gene can significantly reduce the intracellular Met levels in U87 and snb19 cells,while also inhibiting cell proliferation and cell cycle progression.This inhibitory effect may be achieved through MAPK signaling pathway.

Objective To investigate the effect of maternal serum vitamin D (VD)levels on the developing nervous system in in-fants.Methods A total of 1 012 pregnant women's serum VD levels of early pregnancy were given the right amount of VD supple-mentation according to the results.They were divided into 3 groups (normal,deficiency and insufficiency group)according to the third trimester VD levels;and their infants 6-8 months were tested on the developmental quotient (DQ),mental index (MI)and other neurological development index by the Developmental Screening Test for child under six(DST).Results There were up to 94.17% of pregnant women in early pregnancy had severe VD deficiency,the average levels was (16.21 ± 7.54)ng/mL;Although VD levels was increased after supplements,there were 46.25% of pregnant women still deficient in VD.The average DQ score of 1 020 infants was 92.67±16.45 and M1 score was 97.53±17.04,which accorded with normal distribution.The rate of the infants born by mothers who had the normal VD levels were superior to those with VD deficiency and insufficiency on the quotients of out-standing and smart,and there was a significant difference between them(P <0.01).But there was no statistical difference between the deficiency and insufficiency group (P >0.05).The level of VD was positively correlated with DQ and MI,and the correlation coefficients are 0.63 and 0.2(slightly correlated),respectively.Conclusion It should be pay more attention to monitoring the VD levels of the pregnant women during the pregnancy,and for lack should be promptly added.

BACKGROUND:Disequilibrium of proportion of adipogenesis and osteogenesis from bone marrow mesenchymal stem cells (BMSCs)is associated with many bone diseases.However,it has been demonstrated that Wnt signaling pathway could play an important role in regulation of BMSC differentiation.OBJECTIVE:To investigate the different gene expression profiles and to find the target gene on Wnt signaling pathway of the BMSCs,after being induced to osteoblasts and adipocytes respectively using Wnt signaling pathway PCR array.METHODS:The third-passage BMSCs,after being induced to osteoblasts and adipocytes respectively for 7 days.The total mRNA of MSCs was extracted by Trizol.BMSC morphology was observed following osteogenic and adipogenic induction under an inverted microscope.Gene array was detected by rat Wnt signaling pathway PCR array.Non-induction group served as controls.The ratio of increase/reduction gene of osteoblasts and adipocytes was calculated.RESULTS AND CONCLUSION:Under an inverted microscope,BMSCs with high homogenicity were obtained following passage 3.BMSCs differentiated into osteoblasts following osteogenic induction,and into adipocytes following adipogenic induction.Compared with non-induction group,fifteen genes(Dkk1,kremen,FZD1,FZD7,et al.)were expressed up-regulated(ratio ＞ 2)and 16(sFrp 5,β-catenin,Dvl3,Tcf7,et al.)genes down-regulated(ratio ＜ 0.5)when the third-passage BMSCs were induced to adipocytes.Six genes(Dkk1,kremen,β-catenin,Wnt11,et al.)were expressed up-regulated and 15 genes(sFrp5,sFRP4,Fzd1,et al.)down-regulated when BMSCs being induced to osteoblasts.Above-mentioned results suggested that Wnt signaling pathway plays an important role in the osteoblast and adipocyte differentiation from BMSCs.

As one subclass of forkhead proteins, the forkhead box O ( FoxO) transcription factors take part in a series of bio-logical processes including cellular apoptosis, damaged DNA re-pair and cleavage of reactive oxygen species(ROS). Increasing evidence highlights that oxidative stress elicited by FoxOs con-tributes to imbalance of redox status in cells related to bone me-tabolism, resulting in development of the pathogenesis of osteo-porosis. This article reviews the relationship of FoxOs and osteo-porosis, which may be beneficial for the research of pathological mechanism and therapeutic strategy of osteoporosis.

Objective To compare the treatment effect of different doses methylprednisolone(MP)combined with monosialotetrahexosyl ganglioside(GM-1)on experimental spinal cord injury.Methods Based on the duplication of compressed spinal cord animal model,36 rats were divided into three groups randomly:the first group with high dose of MP,the second with moderate dose of MP and GM-1,the third with low dose of MP and GM-1.The motor evoked potential(MEP)and the grades of BBB were detected on the 7th day and 14th day after injury.Results The peak latencies and the amplitudes of MEP and the BBB grades changed significantly after injury in the three groups.After treatment by different administration,the recovery of the two combined groups was more obvious than that of the high dose MP group,and the effect with the low dose of MP and GM-1 group was most significant.Conclusion The treatment effect of low dose MP combined with GM-1 is better than the other two therapies on experimental spinal cord injury.

Objective To investigate the effects of ATP1A1 knockdown by RNA interference(RNAi) on proliferation of human U251 glioma stem cells .Methods The human U251 glioma stem cells were infected with lentivirus expressing ATP1A1-shRNA . The mRNA and protein expressions of ATP1A1 in U251 glioma stem cells were detected by RT-qPCR and Western blotting ,re-spectively .The cell cycle and apoptosis were evaluated by flow cytometry .The proliferation of U251 glioma stem cells was deter-mined by MTT assay .Results The expressions of ATP1A1 in U251 glioma stem cells transfected with ATP1A1-shRNA were in-hibited significantly at both mRNA and protein levels ,with an inhibitory rate of 84 .15% for ATP1A1 mRNA and of 55 .33% for ATP1A1 protein respectively .The proliferation of cells was inhibited ,the cell apoptotic rate was significantly increased and the cell cycle was arrested in G1 phase and S phase decreased significantly in ATP1A1-shRNA cells(P<0 .05) .Conclusion RNAi targe-ting ATP1A1 gene could down-regulates the ATP1A1 expression ,induces cell apoptosis ,regulates cell phase redistribution and in-hibits cell proliferation in U 251 glioma stem cells .

Objective To discuss the feasibility,efficiency and safety of urokinase application in patients with hypertensive intraventricular hemorrhage.Methods Sixty-nine patients were included,30 treated with ventricular drainage alone and 39 receiving adjunctive intraventricular urokinase.CT images and ADL scores were compared between the two groups.Results The intraventricular thrombolysis with urokinase significantly hastened the resolution of intraventricular blood clots as compared with ventricular drainage alone(P=0.030),with better outcome(P=0.029).Conclusion Urokinase application is a simple,effective,and safe in managing hypertensive intraventricular hemorrhage.

Objective To explore the effect of the autoregressive integrated moving average model?nonlinear auto?regressive neural network(ARIMA?NARNN)model on predicting schistosomiasis infection rates of population. Methods The ARIMA model,NARNN model and ARIMA?NARNN model were established based on monthly schistosomiasis infection rates from Janu?ary 2005 to February 2015 in Jiangsu Province,China. The fitting and prediction performances of the three models were com?pared. Results Compared to the ARIMA model and NARNN model,the mean square error(MSE),mean absolute error (MAE)and mean absolute percentage error(MAPE)of the ARIMA?NARNN model were the least with the values of 0.011 1, 0.090 0 and 0.282 4,respectively. Conclusion The ARIMA?NARNN model could effectively fit and predict schistosomiasis in?fection rates of population,which might have a great application value for the prevention and control of schistosomiasis.

Objective To evaluate the effects of metformin on thyrotropin(TSH)levels. Methods From the patients with type 2 diabetes mellitus or metabolic syndrome, 48 patients with primary hypothyroidism were enrolled and grouped. 17 patients were treated only with metformin(group A), 19 patients with metformin and stable L-T4substitution(group B), and the remaining 12 patients with antidiabetic drugs(other than metformin)and L-T4(group C). Meanwhile, 20 euthyroid patients with other thyroid abnormalities(group D)and 30 patients without thyroid diseases(group E)served as control. TSH, FT3, FT4, TT3, TT4, and blood glucose were determined regularly in all these subjects. Results After administration of metformin for 12 months, serum TSH were decreased in group A [(5.05±1.07 vs 2.61±0.91)mU/L, P＜0.01] and group B [(2.67±1.03 vs 1.35±0.74)mU/L, P＜0.01]. No difference was found in FT3and FT4in both groups. TSH levels were raised from(1.30±0.71)to(2.58±1.02)mU/L(P＜0.01)within 8～12 weeks in 13 out of 15 patients after metformin withdrawal. Serum TSH and thyroid hormones in the other 3 groups were not significantly changed. Conclusion Administration of metformin may lead to reduction of serum TSH level.

Objective To investigate the clinical features and molecular characteristics of bacteremia caused by K. pneumoniae and study on the differences between classical and hypervirulent K. pneumonia( hvKP) . Methods The clinical features of 70 cases of K. pneumoniae bacteremia collected from Beijing Chaoyang Hospital from 2008 to 2012 were retrospectively analyzed. The patients were identified according to the records from the Clinical Microbiology Department. Patients′ data were obtained from medical records. The molecular characteristics of the K. pneumoniae strains were also studied using PCR, multilocus sequence typing and pulsed field gel electrophoresis( PFGE) methods. Data were analyzed using the statistical package SPSS for windows version 17. 0. For categorical data, different groups were compared using the Chi-square test to analyze the quantitative variables. P≤0. 05 was considered to be statistically significant. All susceptibility data were analyzed using Whonet, version 5. 6. Results The hvKP was identified from 31. 4% of the patients with K. pneumoniae bacteremia, which displayed 4 serotypes ( K1, K2, K20, and K57). About 40. 9% (9/22) hypervirulent strains were isolated from patients without underlying diseases, while 95. 8% (46/48) of the classic K. pneumoniae infections were found in patients with the presence of one or more underlying diseases. More hvKP positive patients (95. 5%, 21/22) had community-acquired infections compared with classical K. pneumonia (cKP) infected patients (35. 4%, 17/48). There was a significant difference between these two groups (χ2 =21. 912,P <0. 001). Two ESBLs-producing hvKP strains were found. The 22 hvKP isolates had 14 different PFGE patterns, among which 6 isolates ( patients with primary liver abscess) shared similar PFGE patterns. Conclusions The
emerging hvKP was prevalent in patients with severe community-acquired infections in healthy individuals. Two ESBLs-producing hvKP strains were identified. Drug resistance had a rising trend, which called for more attention in clinical settings.