Objective To evaluate the prospects of recombinant 38000 protein of Mycobacterium tuberculosis in tuberculosis epidemic investigation and subunit vaccine preparation.Methods Physicochemical characteristics of recombinant 38000 protein was detected by P I, peptide-mapping analysis and circular dichroism,guinea pig skin test,MTT stain,and peripheral blood macrophage phagocytosis were used to investigate the roles of recombinant 38000 protein in the cell immunity.Results Recombinant 38000 protein was acidic protein,its P I, was 4 67.The number of alkaline amino acid correspond with theoretic number;The secondary structure of recombinant 38000 protein was composed of ?-helix(32 6%),?-turn(31 6%) and random coil(35 8%) Recombinant 38000 protein could induce DTH in guinea pig sensitized by Mycobacterium tuberculosis Recombinant 38000 protein enhanced phagocytosis of macrophage in mice . PBMC from 30 8% healthy donors and 25% tuberculosis patients were stimulated by the recombinant 38000 protein.Conclusion Recombinant 38000 protein may be used as diagnostic reagent and as an candidate in development of subunit vaccine.

Objective To observe the epidemic trends of Keshan disease (KD) from 1967 to 2012 in Chuxiong City,in order to provide a scientific basis for prevention and control of the disease.Methods The data below was collected and analyzed with epidemiological method.KD cases reported through registration and the death case reported were collected in Chuxiong City from 1960 to 2012,also the reported monitoring results of KD and the adult KD screening results of dilated cardiac patients and mountain patients in 2012 were collected,then epidemic trends of KD was analysed comprehensively.Results There were 1 569 cases of KD registered from 1967 to 2012.There were 566 death cases of KD.The total prevalence rate of KD was 7.7％ (226/2 953) by monitoring children KD at the diseased areas of KD and the potential KD patients accounted for 97.3％ (220/226) from 1990 to 1994.The total prevalence rate of KD was 0.6％ (44/7 174) by surveillance at KD diseased areas from 2005-2012 and the potential KD patients accounted for 70.5％ (31/44),there were 7 children cases of KD under 15 years who accounted for 15.9％ from 2005 to 2012.Totally 313 551 children were given sodium selenium for supplying selenium from 1976 to 2008.Totally 151 cases of KD were found by screening from the patients with dilated cardiac patients since 1988 and they were all chronic KD patients.There were 112 cases of KD in 2012 and there were 6 children cases of KD under 15 years,which accounted for 5.4％.Chuxiong City Keshan disease epidemic presented from subacute Keshan disease to Keshan disease,to latent Keshan disease a slow transition;age from children (＜6 years) to children (＜15 years) to adult-oriented features.Conclusions The incidence and prevalence rate of KD has reached the basic control standards,but children KD is coexisting with adults KD.The pathogenic factors have not been eliminated,disease monitoring and health promotion should be carried out.

Objective To investigate the potential role of tumor necrosis factor receptor-associated factor-6(TRAF6)in the rat cerebral ischema-reperfusion injury.Methods 40 healthy adult SD rats were divided into 5 groups(n=8)according to the ran-dom control principle:sham operation group,ischemia group,reperfusion 2 h group(R2 h),reperfusion 12 h group(R12 h)and reperfusion 24 h group(R24 h).The rat cerebral ischemia-reperfusion injury model was constructed.The change of TRAF6 expres-sion was examined by RT-PCR and Western-blot.Then,the immunohistochemistry was adopted to locate the TRAF6 protein.Re-sults Compared with the sham group,the expression of TRAF6 in the ischemia group and the R2 h,R12 h and R 24 h groups was obviously increased,but the difference had no statistical significance (P <0.05).TRAF6 was mainly located in the cytoplasm of neuronal cells.Conclusion Activated TRAF6 is involved in the brain cell death induced by cerebral ischemia-reperfusion.

Objective To compare expression of extracellular proteins of virulent H37Rv and attenuated H37Ra in order to search differential proteins,to provide a train of thought for studing M.TB toxicity further.Methods Extracellular proteins were extracted from H37Rv and H37Ra which were inoculated and cultured on Suton's medium for three weeks.The first dimensional electrophoresis was performed on immobilized pH gradient rod gels(pH 3～10).Then the proteins in the rod gels were separated using SDS-PAGE gels.The silver-stained gels were dried and scanned with image scanner.The 2D image analysis was performed with image Master 2D Elite 3 10.Results The most protein spots deriving from extracellular proteins of H37Rv and H37Ra strains were in acidic range.In the basic range(pI more than 9 0),the number of protein spots belong to extracellular proteins of H37Rv and H37Ra was few.Three protein spots belong to low molecular range in H37Rv strain.However,absent in H37Ra strain.Conclusions Two-dimensional gel electrophoresis is useful to separate protein in Mycobacterium tuberculosis.

Objective To understand the rpoB gene mutation in M.tuberculosis isolates,and to evaluate their clinical value.Method 335 clinical isolates of mycobacterium tuberculosis(109 isolates drug susceptible,246 isolates rifampin-resistance or multidrug resistance including rifampin) were detected using polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP).Results SSCP pattern of reference mycobacterium tuberculosis H37Rv as control,no mutation was found in rifampin-suscepitible 109 strains.SSCP patterns of 225/246 rifampin resistant clinical isolates were different from the normal control.The sensitivity was 91 5%.31 resistant isolates,included 25 abnormal isolates and 6 normal isolates of SCCP were identified.Sequencing showed 29 isolates had rpoB gene mutations and 3 isolates were not found rpoB gene mutations.The 3 most frequent rpoB gene mutation situs were Leu-531(19 isolates.TCGTTG) and His-526(7 isolates,CACTAC) and Asp-516(3 isolates,GACGTC).Conclusions The results confirm that rpoB gene mutation is the most important mechanism in rifampin resistant tuberculosis mutation situs are 531 tryptophan and 526 histidine;respectively.It is feasible that using PCR-SSCP to detect drug resistance in mycobacterium tuberculosis.

Objective Investigate the expression difference of miRNA in cerebrospinal fluid (CSF) to explore new biomarkers for glioma diagnosis and evaluate the diagnostic value .Methods The candidate biomarkers in CSF were detected by using FQ-PCR for 20 cases of glioma patients and 20 cases of non-glioma patients(control group) .miRNAs with significant level changes in CSF sam-ples from patients with gliomas (n=20) compared with healthy volunteers (n=20) were screened out by using Mann-Whitney U test and Kruskal-Wallis test ,and the diagnostic values were evaluated by receiver-operating characteristic curves (ROC curves)and area under the curve(AUC) .Results MiR-128 and miR-21 were differentially expressed in CSF samples from patients with gliomas compared with control group .Expression of miR-21 in glioma is significantly higher than that in the control group(P<0 .05) ,while the expression of miR-128 in gliomas was significantly lower than that in control group(P<0 .05) .AUC was 0 .96 when using only miR-21 as the diagnostic biomarker ,and the sensitivity was 90% ,specificity was 95% .The diagnostic sensitivity and specificity were 100% when MiR-128 and miR-21 combined .Conclusion miR-128 and miR-21 are potential markers for gliomas diagnosis in the CSF .

Objective To retrospectively demonstrate risk factors for mortality in multiple trauma patients with acute respiratory distress syndrome(ARDS).Method This wes a retrospective cohort stuay regarding multi-ple trauma as a single cause for intensive care unit admission.Patients identified multiple trauma with ARDS en-rolled in prospectively maintained database between May 2003 and April 2008 were observed,and 23 items of po-tential risk factors of impacting mortality were calculated by univariate and multivariate logistic analyses in order to find distinctive items in these multiple trauma patients.Information on patients demographics characteristics,treat-ment procedures and injury severity were collected at the time of EICU admission.The criteria used for ARDS met definition of the guideline(2006)of Chinese medical association.The commonly accepted definition of multiple injuries was consistent with both several injury sites(generated from two or more than two anatomic sites)and in-jury in one anatomic site at least threatening life.Severity of injury was quantified by injury severity seore and the simplified acute physiology score and chronic health evaluation score (APACHE Ⅱ)in EICU admission.We in-cluded adult patients(age≥18 years),those with an EICU length of stay longer than 48 hours,and those accept-ing mechanical ventilation more than 24 hours.Patients who were readmitted to EICU by virtue of non-traffic injury or transferred to EICU from other hospitals after long-term treatment were excluded.Mortality was assessed at the 28th clay after trauma.Results There were 269 multiple trauma patients with posttranmatic ARDS admitted to ICU during the study period,the unadjusted odds ratio(OR)and 95% confidence intervals(CI)of mortality were associated with six risk factors(APACHE Ⅱ score,duration of tratuna factor,pulmonary contusion,aspiration of gastric contents,sepsis and duration of mechanical ventilation)out of 23 items.The adjusted Odds Ratios(ORs) with 95% CI were denoted with respect to surviving beyond 96 hours ICU admission(APACHE Ⅱ score,duration of trauma factor,aspiration of gastric contents),APACHE Ⅱ score beyond 20 ICU admission(duration of trauma factor,scpsis,duration of mechanical ventilation)and mechanical ventilation beyond 7 days ICU admission(dura-tion of trauma factor and sepsis).Conclusions Impact of pulmonary contusion and APACHE Ⅱ score contribut-ing to prediction of mortality may exist in prophase after multiple trauma.Sepsis is still a vital risk factor referring to systemic inflammatory response syndrome,infection,and secondary multiple organs dysthnetion.Aspiration of gastric contents could lead to incremental mortality due to scvere ventilation associated pneumonia.Duration of trauma factor determined degree of injury and outcomes,longer duration generally manifested higher mortality.Long-standing mechanical ventilation should be constrained on account of occurring severe refractory complications.

Osteopontin (OPN) is a secreted O-glycosylated phosphoprotein that exists in a variety of tissues and body fluids, with a variety of biological activity. Integrin α_vβ_3 is the main receptor. OPN mainly involves in cell proliferation, differentiation, migration and adhesion. The study of OPN at home and abroad mainly focuses on the bone resorption, angiogenesis, atherosclerosis, digestive system, urinary system, wound healing, skin fibrosis, liver fibrosis, kidney fibrosis, etc.But reports about OPN in pulmonary fibrosis are much less, now the relationships between OPN and pulmonary fibrosis are reviewed.

Objective To investigate the imaging and clinical features of Percheron artery infarction.Methods The clinical manifestations,imaging data,treatment,and prognosis of 10 patients with Percheron artery infarction were analyzed.Results All 10 patients had acute onset and had different degrees of consciousness disorders,including vertical gaze palsy (n =3),language disorder (n =3),limb weakness (n =6),and memory impairment (n =4).MRI showed bilateral thalamus infarction.Midbrain V-sign in one patient was observed.Cerebral angiography revealed that one patient had Percheron artery infarction.All 10 patients were improved and discharged,but left with varying degrees of sequelae.Conclusions Percheron artery infarction is rare in clinical practice.The diagnosis is mainly based on clinical manifestations and imaging examinations.Cerebral angiography can diagnose the disease.

Objective To investigate the expression changes and significance of Pdlim2 in the glomerular podocyte of hyperlipidemic rats.Methods Forty-five individuals of SD rats were divided randomly into 3 groups (n =15 in each group).The control group was fed with normal diet.The high fat group was fed with high fat diet.The simvastatin group was fed with high fat diet plus with simvastatin gavage (10 mg· kg-1 · d-1).Five rats were sampled randomly from each group at week 4,6,and 10 and the urinary protein excretion,the concentration of serum cholesterol,and the concentration of low density lipoprotein cholestorol were determined,the glomerular podocyte damage in rats was detected by electron microscope,the expression of Pdlim2 protein was determined by immunohistochemistry and by Western blotting.Results The levels of serum cholesterol and low density lipoprotein cholestorol increased significantly in high fat group and simvastatin group at week 4 compared to that in control group(P ＜ 0.05),and the level in simvastatin group was significantly decreased compared with that in high fat group(P ＜ 0.05).The urinary protein levels of high fat group and simvastatin group were significantly higher than that in control group at week 10,and the level in simvastatin group was significantly decreased in high fat group,there was significant difference in each group of comparison(P ＜ 0.05).Podocyte injury was detected by electronic microscopy in high fat group at week 4,and the injury became more serious as the treatment time increased.Podocyte injury in the simvastatin group was significantly less than that in the high fat group and the control group at week 10.The positive staining of Pdlim2 was mainly in the glomeruli and the expression of Pdlim2 of the high fat group was lower than the simvastatin group,and both were lower than the control group at week 10.The expression of Pdlim2 protein of high fat group was lower than that in the control group since week 4(P ＜0.05).The expression of Pdlim2 protein of high fat group was lower than that in the simvastatin group (P＜0.05),and both were lower than the control group at week 10(P＜0.05).Conclusions Hypedipidemia induces podocyte injury before urinary protein,which is suggested to be associated with the decrease of Pdlim2 protein.Simvastatin reduces podocyte toot processes of high fat induced fusion,which may be through protecting the expression of glomerular Pdlim2.