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.

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 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.

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.

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.

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

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.

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.

Background Posterior chamber phakic intraocular lens implantation is a safe and effective method for correction of high myopia.Petacam-assisted observation of the intraocular lens position in the eye after surgery,provides a good way for the evaluation of operation safety.Objective This study was to evaluate the security and effectiveness of posterior chamber (phakic) implantable contact lens (ICL) implantation for high myopia.Methods Twenty-five eyes of eighteen high myopia patients who received ICL implantation were retrospectively analyzed from March 2009 to September 2012.The mean preoperative spherical equivalent (SE) of the patients was (-17.52 ± 1.73) D and the cylinder was (-0.75 ± 0.28) D.The clinical effective indexes included preoperative and postoperative uncorrect visual acuity (UCVA) and best correct visual acuity (BCVA),SE,intraocular pressure (IOP),corneal endothelial cell number and percentage of corneal endothelial hexagon cells.In addition,Pentacam analysis system was used to compare the anterior chamber depth,anterior chamber angle and space between crystal lens and ICL between before and 6 months after operation.Written informed consent was obtained from each patient before any medical procedure.Results ICL was successfully implanted in all the 25 eyes.UCVA was 1.47 ±0.26 and 0.26±0.15 respectively before and after operation,showing a significant difference between them (t =21.71,P =0.00).SE,corneal endothelial cell number and percentage of corneal endothelial hexagon cells were significantly lower than those before operation (SE:t =-48.60,P =0.00 ; corneal endothelial cell number:t =13.07,P =0.00 ;percentage of corneal endothelial hexagon cells:t =10.79,P＜0.05).Anterior chamber angle was (25.02± 4.77)° 6 months after operation and was smaller than (38.43±4.04)° before operation (t=15.32,P=0.00).No significant differences were found in IOP and anterior chamber depth between before and after operation (IOP:t =-1.57,P =0.13 ; anterior chamber depth:t =1.46,P =0.16).The spaces between crystal lens and IOL was (363.33± 44.37) μm 6 months after operation,with a significant decline in comparison with (542.17±39.46) μm 1 week after operation (t =20.86,P =0.00).Glare sense appeared in 2 eyes after operation.Conclusions The results indicated that ICL implantation can retain the natural accommodation of lens.ICL implantation appears to be an effective and safe approach to high myopic eyes.However,the changes of corneal endothelial cells after operation should be payed more attention.

This article gave a detailed account of the practice and characteristics of medical support training in U.S. Navy hospital ship MERCY.We compared the details of medical support training in this ship with those in the Chinese Na-vy hospital ship Ark Peace.We offered suggestions as follows:①During the future medical support training on board, we should regard training procedures and medical techniques as equally important.②We should establish an examination and evaluation team for the medical staff during the training.③We should establish selection standards for significant missions and conduct routine evaluation of the medical staff aboard the hospital ship.