Objective To observe the cell proliferation inhibition of DNA methyltransferase (DNMT) inhibitors decitabine (DAC) combined with daunorubicin (DNR) in human leukemia cell line HL-60.Methods The effects of DNR and DAC were examined in HL-60 cells by cell viability using MTT method,and cell death using flow cytometric (FCM).Results DAC,DNR single drug application showed their effects on cell proliferation was dependent of dose and time,the inhibition effect of combined treatment group was much clearer [inhibition ratio of 72 hours was (80.23±1.71) ％,P ＜ 0.001].The highest apoptosis rate was at 5.0 μmol/L DAC combined with 1.0 μmol/L DNR for 72 hours,which was statistic significant (F =30.199,P ＜ 0.001).Combinations of different concentrations of DAC and DNR increased expression of PTEN mRNA in concentration-dependent manner,which was significantly higher than the control group and DNR single drug group (F =578.218,P ＜ 0.001).Conclusions DAC can significantly inhibit the proliferation of HL-60 cells and induce apoptosis,synergistic effect can be observed when DAC combined with DNR.The underlying mechanism can be due to DAC demethylation effect to increase PTEN mRNA expression.

Objective Based on dual channel melting curve analysis-based assay,we developed a method to rapidly detect the drug-resistant mutations in Mycobacterium tuberculosis through real-time PCR.Methods According to the common first-line drug-resistant mutations of Mycobacterium tuberculosis,we designed six dual-labeled fluorescence probes to rapidly detect the drug-resistant mutations through realtime PCR melting curve after amplifications of drug-resistant related gene region of DNA.The targets include rpoB 81 bp core region,katG315,inhA promoter,ahpC promoter and embB306.To validate the sensitivity and specificity of our method,we performed real-time PCR assays to detect drug-resistant mutations in 76 clinical MDR-TB samples,which were collected by Shanghai CDC in 2008.Results In the validation,this method successfully detected drug-resistant mutations in all 76 clinical MDR-TB samples.The △Tm of mutations were from 1.8 to 14.4 ℃.Comparing with the sequencing data,all mutations covered by the six probes were detected with 100％ sensitivity and 100％ specificity (rpoB,80/80; inhA,7/7 ; katG315,59/ 59;ahpC,8/8;embB306,27/27).This method can successfully detect drug-resistant mutations from 100 copies/μl DNA samples.Conclusions A widely applicable real-time PCR assay to detect first line drug-resistant mutations of Mycobacterium tuberculosis has benn developed.This method has proven to have the advantages of high sensitivity,specificity and low risk of contamination.It can be used in rapid diagnosis of clinical drug-resistant tuberculosis and the evaluation of laboratory drug sensitivity test.

DEC1 is a basic helix-loop-helix(bHLH)transcription factor involved in the regulation of cell prolif-eration;cell differentiation;lymphocytes maturation;circadian rhythms;immune response and response to hypoxia.Recent studies have revealed that the expression of DEC1 is abnormally high in various tumor tissues and cells.In addition;the expression of DEC1 in the tumor tissue is related to the malignancy of various cancer types.This paper;therefore;focuses on the relationship between DEC1 and proliferation;apoptosis;cellular senes-cence;invasion and metastasis of tumor cells;in order to elaborate the role of DEC1 in the pathogenesis and pro-gression of tumor.

Electroencephalography (EEG) is an important instrument for the evaluation of brain function, and an irreplaceable diagnostic technique for nervous system diseases. At present, China still lacks professional child-EEG talents. Therefore, it is a task of great priority to establish an effective and practical training method and foster more child-EEG physicians. As most trainees have not learned EEG before and only have limited time for learning, we divide the child-EEG training into three phases, includ-ing theory learning, practice training, and EEG reading and interpretation on the basis of the general rules in learning EEG. In the theory learning phase, basic EEG knowledge is taught comprehensively to form a solid foundation for future study. In practice training phase, the trainees acquire important skills of EEG by carrying out complete EEG monitoring, eliminating EEG artifacts, observing seizures, and read real-time EEG. In the phase of EEG reading and interpretation, the trainees learn to analyze EEG gradually by read-ing and report EEG under the guidance of the senior physician. Strict examination is arranged for each phase to evaluate study results objectively. The phased model is designed to implement a step-by-step training of child-EEG and foster the trainee's independent ability to carry out EEG inspection.

Objective To investigate the expression of Caspase-9 and heat-shock protein-90 (HSP 90) in rats after focal cerebral ischemia-reperfusion injury.Methods The male SD rats (200-250 g) were divided into three groups by the random number table: normal group, sham group and cerebral ischemia-reperfusion (CIR) group.Each group was sorted into four subgroups including group 6 h, group 24 h, group 48 h and group 72 h according to the reperfusion time.Suture-occluded method was adopted to prepare focal cerebral ischemia-reperfusion(CIR) injury in rat model.Enzymelinked immunosorbent assay (ELISA) method was used to detect the variations of Caspase-9 and HSP-90 expression in rats.Results The changes in Caspase-9 and HSP 90 expression in the brain cells were observed by ELISA method.The expression of Caspase-9 and HSP-90 was weakly expressed in sham group, and was at peak in CIR group within 24 h-48 h, then began to decline at 72 h after the reperfusion time.The differences in the expression of caspase-9 and HSP-70 between sham group and normal group were not statistically significant.Conclusions Apoptotic cells gradually increase along with reperfusion time and reach the peak at 48 h after cerebral ischemia-reperfusion.In ischemia half dark stripe, the expression of Caspase-9 and HSP 90 is increased in neuronal cells after cerebral ischemia-reperfusion, and the positive cells number is at peak at 48 h after cerebral ischemiareperfusion.Apoptosis of neuronal cells after cerebral ischemia and reperfusion is a dynamic evolutionary process.The expression of Caspase-9 and HSP 90 in nerve cells plays an important role in regulating cell apoptosis.

Objective To find out the resistant situation and drug of Mycobacteria patients in Sichuan and offer foundation for clinical.Methods Two hundred randomized clinical isolates of Mycobacterium were determined by Roche drug sensitivity and minimum inhibitory concentration (MIC) method.Results Of the 200 clinical isolates,192 stains were Mycobacterium tuberculosis(MTB) (96.0％),8 strains (4.0％) were non-tuberculosis mycobacterium(NTM).Of the 192 MTB strains,108( 57.3％ ) sensitive strains and 84 (43.7％)stains were resistant to one or more than one drugs.Among these 84 resistant strains 23 were multi-drug resistant ( MDR,12.0％ ),4 were extensively drug resistant( XDR,2.1％ ).The anti-TB drug resistance rates were:SM(16.7％),INH(20.8％),RFP(17.2％),EMB(10.9％),PI(16.1％),LFX(8.8％),AMK ( 16.7％ ),CPM ( 6.2％ ),PTA ( 33.3％ ),respectively.Conclusion The resistance rate of tuberculosis keeps at a high level in Sichuan,especially the resistance rate of multiple (≥4) drug,we should oar attention.

Aim at the two problems in the field of traditional Chinese medicine (TCM) mechanism elucidation, one is the lack of detailed biological processes information, next is the low efficient in constructing network models, we constructed an auxiliary elucidation system for the TCM mechanism and realize the automatic establishment of biological network model. This study used the Entity Grammar Systems (EGS) as the theoretical framework, integrated the data of formulae, herbs, chemical components, targets of component, biological reactions, signaling pathways and disease related proteins, established the formal models, wrote the reasoning engine, constructed the auxiliary elucidation system for the TCM mechanism elucidation. The platform provides an automatic modeling method for biological network model of TCM mechanism. It would be benefit to perform the in-depth research on TCM theory of natures and combination and provides the scientific references for R&D of TCM.
Animals ; Automation ; instrumentation ; methods ; Databases, Factual ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Gene Regulatory Networks ; Humans ; Medicine, Chinese Traditional ; Plants, Medicinal ; chemistry

A novel antimicrobial peptide, named as perinerin (GenBank accession No. P84117), was isolated and characterized from Asian marine clamworms, Perinereis aibuhitensis Grube. Perinerin showes powerful and broad activity against both grampositive and gramnegtive bacteria in vitro, especially on Pseudemonas aeruginosa. To obtain large amounts of active perinerin and characterize its main physiochemical features, The perinerin weve expressed in Pichia pastoris. Intact perinerin gene amplified by the modified gene SOEing method(Gene splicing by overlap extension)was cloned into expression vector pPICZ?A and obtained recombinant vector pPICZ?APEN, then pPICZ?APEN was expressed in the Pichia pastoris GS115. The expressed sample was analyzed by TricineSDSPAGE. The results showed that Pichia pastoris was a suitable system producing the secreted form of perinerin. Bioactivity assay showed that the recombinant perinerin had marked antimicrobial effects.

Objective To investigate the effectiveness of disodium Aidi injection for cancer-related fatigue in nasopharyngeal carcinoma patients of Ⅲ-Ⅳ B stage undergoing radiotherapy and chemotherapy.Methods Eighty nasopharyngeal carcinoma patients of Ⅲ-Ⅳ B stage with fatigue symptoms from December 2011 to May 2012 in our hospital were divided into two groups.All patients received treatment of sequential 3 cycles with platinum-based chemotherapy after concurrent chemoradiation.One group of 40 patients also received intravenous infusion of disodium Aidi injection (experimental group),the other group of 40 patients only received conventional therapy (control group).Brief fatigue inventory (BFI) questionnaires data were collected at baseline,the eighth week and the twentieth week after treatment.The changes of fatigue severity and the occurrence of Ⅲ-Ⅳ degree adverse reactions in the two groups were compared.Results At the eighth week,the improvement in fatigue severity was not significantly different between two groups (x2 =1.758,P =0.32).However,significant improvement in cancer-related fatigue of experimental group was found than that of control group at the twentieth week(x2 =8.12,P =0.005).The Ⅲ-Ⅳ degree adverse reactions of experimental group were significantly lower than that of control group.Conclusion Disodium Aidi injection combined with radiotherapy and chemotherapy can improve the cancer-related fatigue of nasopharyngeal carcinoma patients of Ⅲ-ⅣB stage and it can also reduce the incidence rate of Ⅲ-Ⅳ degree adverse reactions.

Aim To screen a more suitable transfection recep-tor,and improve the efficiency of constructing cell lines highly expressing human peptide transporters 1 (hPepT1 ).Methods The recombinant plasmid pcDNA3.1 (+)-hPepT1 was transfect-ed into MDCK cells and HeLa cells by LipofectamineTM 2000 transfection reagent,respectively.The monoclonal cells were se-lected and cultured.Expression of hPepT1 mRNA and protein were determined by qRT-PCR and Western blot,respectively. The uptake capacity of Glysar in transfected cells was examined. Results Compared with wild type cells,the expression of hPepT1 and the uptake of Glysar in transfected MDCK cells and HeLa cells significantly increased (P <0.05).Although the up-take of Glysar in HeLa cells was higher than that of MDCK cells,on the contrary,the expression of hPepT1 and the uptake of Glysar in MDCK-hPepT1 cells was higher than that of HeLa-hPepT1 cells.Conclusion MDCK cells may serve as a more suitable transfected receptor for the construction of a cellular model with high expression of hPepT1 ,which would make the construction of a cell model highly expressing hPepT1 more effi-cient.