Objective To establish Phage Amplified Biologically Assay (PhaB) detecting isoniazid(INH) resistance rapidly and evaluate PhaB assay for drug susceptibility testing of isoniazid(INH) in clinical isolates of Mycobacterium tuberculosis(MTB). Methods Detecting the INH resistance of 167 clinical isolates of MTB by PhaB assay,comparing the results of PhaB with that of Bactec-960 system and analyzing the sensitivity, specificity and accuracy of PhaB assay. Results When the mixture of 0.2 ?g /ml INH and MTB was incubated in 37℃ for 48 h, the accurate results were obtained rapidly by calculate the reduce of plaque of PhaB assay. If the results of Bactec-960 system is the golden standard, the sensitivity, specificity, PPV, NPV and accuracy of PhaB assay was 96.4%, 96.4%, 93.1%, 98.2% and 96.4% respectively. Conclusions The PhaB assay with highly sensitivity specificity are highly consistent with Bactec-960 system. Not only it takes only three days to detect drug susceptibility of INH in clinical isolates of MTB but also it is easily to operate. We believe that this low-cost assay may be a good rapid screening of INH resistance in MTB isolates.

Objective Evaluating possibility of phage amplified biologically(PhaB) assay in detecting susceptibility of Mycobacterium tuberculosis(MTB) to four first-anti-tuberculosis-drugs on the same time and of 139 clinical isolates to streptomycin(S), isoniazid(H), rifampin(R) and ethambatal(E) at the same time, comparing with the results of Bactec-960 and determining the minimal inhibitory concentrations(MIC) of isolates which results were not consistent. Results Concordance rates of the susceptibility to S, H, R and E in 139 clinical isolates detected by PhaB and Bactec-960 are 97.1%, 99.3%, 95.7% and 95.0% respectively. If the results of Bactec-960 system is the golden standard, the sensitivity, specificity, positive and negative predictive value (PPV and NPV) as well as accuracy of susceptibility test to S detected by PhaB assay was 90.0%, 99.1%, 96.4%, 98.2% and 97.1% respectively, to H 97.9%, 98.9%, 97.9%, 98.9% and 95.0% , to R 86.2%, 97.3%, 89.3%, 96.4% and 95.0%, to E 81.0%, 97.5%, 85.0%, 96.6% and 95.0%. There are 19 inconsistent results of 13 isolates in comparing PhaB with Bactec-960. 18 results of 12 isolates by MIC are identical with the results of PhaB assay. 1 result of 1 isolate is identical with Bactec-960. Conclusions[KG1]The results of susceptibility to S, H, R and E detected by PhaB were highly concordance rate with the results of Bactec-960. PhaB assay can be used for rapid screening of susceptibility test for MTB.

Objective To set up and evaluate the method of Phage Amplified Biologically Assay (PhaB) in the rapid detection of detection of Ethambutol resistance. Methods To detect the EMB resistance of 102 clinical isolates of Mycobacterium tuberculosis by PhaB and compare it with the results of absolute concentration method. The minimum inhibitory concentration (MIC) was detected for all discrepancy isolates. Results Of all 102 strains in Mycobacterium tuberculosis clinical isolates, 82 strains were EMB susceptible and 20 strains were EMB resistant by PhaB method, while 77 strains were EMB susceptible and 25 strains were EMB resistant by absolute concentration method. 74 of 102 strains were EMB susceptible and 17 strains were EMB resistant by both methods. The concordant isolates of determination of EMB resistance were 91 strains in two methods and the concordance rate was 89.2%. There were 11 disconcordant isolates and the discrepancy rate was 10.8%. In the 11 strains of discrepant isolates between two methods, 7 strains (63.6%) were in accord with the results of MIC method (5 of 7 strains were EMB susceptible by PhaB but EMB resistant by the absolute concentration methods, 2 of 7 strains were EMB resistant by PhaB but EMB susceptible by the absolute concentration method). Conclusions The PhaB assay can be used for detection of EMB resistance in isolates of Mycobacterium tuberculosis easily and quickly within three days.This method do not need special instrument and may be used for rapid screening of M.tuberculosis with resistance to EMB.

Purpose To investigate the effects ofβ-1apachone on inhibition of pro1iferation and migration and induction of apoptosis in gastric cancer ce11s in vitro. Methods The ce11 viabi1ity was detected using MTT and co1ony formation assay,the migration abi1ity was determined using scratch assay method,and the apoptosis was examined using f1ow cytometry. Meanwhi1e,the expression of biomarkers of pro1iferation,EMT markers andapoptosiswere detected using Western b1ot ana1ysis. Results β-1apachone cou1d significant1y inhibit the pro1iferation of SGC-7901 and AGS gastric cancer ce11s( P<0. 05),and down-regu1ate the expression 1eve1s of Skp2 and DEK pro-teins. β-1apachonecou1d a1so inhibited the invasion and moti1ity of gastric cancer ce11s via down-regu1ating the expression 1eve1s of MMP-2/9 and Ezrin proteins and up-regu1ating the epithe1ia1 markers. In addition,β-1apachone enhanced the apoptosis of gastric canc-er ce11s,down-regu1ation of BCL-2/Bax ratio and up-regu1ation of activated Caspase-3/8/9. Conclusions β-1apachone can effective1y inhibit the pro1iferation and induce the apoptosis of gastric cancer ce11s,and inhibit the migration of gastric cancer ce11s via MMPs and EMT pathways.

Objective:To observe influence of up‐regulated expression of myocardial heat shock protein (HSP) 70 in‐duced by heat stress on myocardial calcium‐activated potassium channel (KCa ) 3.1 expression in rabbits with atrial fibrillation (AF) caused by rapid atrial pacing (RAP) .Methods :A total of 24 New Zealand white rabbits were ran‐domly divided into sham operation group (n=8 ,only implant electrode without pacing ) ,pacing group (n=8 ,right atrium (RA) received RAP at 600 times/min for 6h) and heat stress pacing group (heat stress group ,n=8 ,received heat stress preconditioning ,then the same RAP as pacing group ) .Results:Compared with sham operation group and pacing group ,there were significant up‐regulation of HSP70 mRNA and protein expression in different sites of heart [HSP70 protein ,left atrium (LA):(39.00 ± 3.21) vs .(39.75 ± 2.82) vs .(69.75 ± 3.45) ,RA: (38.38 ± 2.92) vs .(39.50 ± 3.89) vs .(69.00 ± 2.93) ,left atrial appendage (LAA):(37.75 ± 3.28) vs .(39.00 ± 3.89) vs . (68.63 ± 3.23) ,right atrial appendage (RAA): (37.00 ± 3.85) vs .(38.38 ± 3.74) vs .(68.75 ± 2.82)] in heat stress group , P<0. 01 all ,but there were no significant difference between pacing group and sham operation group , P>0.05 ;compared with pacing group with down‐regulation of KCa3.1 mRNA and protein expressions ,there were significant up‐regulation of KCa3.1 mRNA and protein expressions in different sites of heart [KCa3.1 protein ,LA:(21.25 ± 1.67) vs .(24.00 ± 2.62) ,RA :(21.13 ± 1.96) vs .(23.75 ± 1.83) ,LAA :(21.00 ± 2.07) vs .(23.75 ± 1.67) ,RAA:(20.88 ± 2.03) vs .(23.50 ± 2.45)] in heat stress group ,P<0.05 all ,and there were no significant difference between heat stress group and sham operation group , P>0. 05. Conclusion:Heat stress may induce up‐regulated expression of myocardial HSP 70 of myocardium ,and HSP 70 may inhibit down‐regulation of KCa 3. 1 mR‐NA and protein expressions in rabbits with atrial fibrillation.

OBJECTIVE To set up and evaluate the microscopic observation drug susceptibility assay(MODS)technology and use it to detect rapidly Mycobacterium tuberculosis(MTB).METHODS The 24-hole cell culture plate method of liquid culture were used to set up MODS technology.The MODS technology was used to detect MTB and non-M.tuberculosis(NMT)isolates comparing with Lowenstein-Jensen(L-J) method.RESULTS When bacterial concentration was 3?103 CFU/ml,the time of reading test-result was the seventh day by MODS;four kinds of NMT(M.phlei,M.kansasii,M.chelonaeand M.marinum) in the liquid medium in the observation was similar to the growth of cording,it was difficult to distinct from the cording of MTB in the liquid medium;When using the 4-Nitro-benzoic acid 800 ?g/ml,thiophene-2-carboxylic acid hydrazine 2.5 ?g/ml for testing conditions,the correct diagnosis can be improved;the test results of clinical isolates by MODS were highly concordance rate with the results of L-J.If the results of L-J was the golden standard,the sensitivity,specificity,positive and negative predictive value as well as accuracy by MODS was 95.7%,100%,100%,99.9% and 97% respectively.CONCLUSIONS The results of MODS in detection of MTB are highly concordance wih the results of L-J method;MODS assay can be used for rapid detection of tuberculosis,with a rapid,simple,inexpensive,and other advantages.

Objective:To explore influence of thrombus aspiration combined bivalirudin during emergency PCI on my- ocardial tissue perfusion and clinical prognosis in patients with acute ST elevation myocardial infarction (STEMI). Methods:A total of 102 patients with acute STEMI,who were confirmed with thrombus burden by CAG in our hos- pital from Jan 2012 to Jun 2014,were selected.According to random number table,they were randomly divided into thrombus aspiration + bivalirudin group (n=52,thrombus aspiration group)and heparin group (n=50,routine PCI group).TIMI blood flow grade 3 rate,TIMI myocardial perfusion grade (TMPG)after PCI,ST segment re- gression rate 2h after PCI,peak value and peak time of cTnI after PCI,LVEF,LVEDd,incidence rates of bleeding and major adverse cardiovascular events (MACE)on one week and one month after PCI were compared between two groups.Results:Compared with routine PCI group,there were significant rise in postoperative TMPG grade 3 rate (56.00% vs.88.46%),TIMI grade 3 rate (58.00% vs.88.46%)and ST segment regression rate (52.00% vs. 76.92%)in thrombus aspiration group,P<0.05 or <0.01. Compared with routine PCI group one month after PCI,there was significant rise in LVEF [(53.76±5.24)% vs.(57.95±5.51)%],and significant reductions in LVEDd [(53.70±3.39)mm vs.(50.63±1.24)mm],peak value [(16.00±4.28)μg/L vs.(13.81±4.00)μg/L]and peak time [(14.00±2.80)h vs.(13.00±2.23)h]of cTnI in thrombus aspiration group,P<0.05 or <0.01. Incidence rate of mild bleeding in thrombus aspiration group was significantly lower than that of routine PCI group (1.9% vs.16.0%),P<0.05,but there was no significant difference in incidence rate of MACE between two groups,P>0.05. Conclusion:Thrombus aspiration combined bivalirudin during emergency PCI is safe and fea- sible for acute STEMI patients,it can effectively reduce incidence rate of bleeding,remove coronary thrombus,im- prove myocardial tissue perfusion and doesn't increase incidence rate of MACE.

Objective To construct the recombinant plasmid of protein CFP10-MPT48-TB8.4 of Mycobacterium tuberculosis and to investigate the diagnosis potential of this fusion protein in tuberculosis serodiagnosis.Methods The recombinant fusion protein CFP10-MPT48-TB8.4 was expressed, and identified by Western blot.The ELSIA based on the purified fusion protein was done,and used for screening in 230 cases of clinical serum samples including pulmonary tuberculosis patients ( n =150 ),pulmonary disease patients other than tuberculosis (n =70) and health controls (n =103 ).The test result was analyzed by Medcale11.5 software.Results The fusion protein CFP10-MPT48-TB8.4 was successfully expressed with a purity over 95％.Specific immunogenicity of the recombinant protein was confirmed by Western blot.The overall sensitivity and specificity obtained of ELISA were 56.7％ (85/150) and 90.8％ ( 157/173 ),respectively.The specificity was 85.7 ％ (60/70) in non-tuberculosis group and 94.2％ (97/103 ) in healthy group,respectively.Conclusion The recombinant protein of CFP10-MPT48-TB8.4 has a high sensitivity and specificity and may be a potential candidate antigen in tuberculosis serodiagnosis.

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 study the relationship of two variants( -871A/G and -336A/G) polymorphisms of the DC-SIGN gene with the susceptibility to pulmonary tuberculosis in Chinese population.Methods Two hundred and thirty-seven tuberculosis cases and 244 controls were genotyped by pyrosequencing in this case-control study. The analysis of the relationship of the -871A/G and -336A/G polymorphisms with their susceptibility of pulmonary tuberculosis(PTB) and the relationship of the two variants with their clinical correlation of tuberculosis was performed by chi-square test. Results The genotypic frequencies of A/G + G/G and A/A of - 871, 37.6%, 62.4% respectively in cases, and 43.4%, 56. 6%respectively in controls, had no significant difference in statistics. And the genotypic frequencies of A/G + G/G and A/A of -336, 12. 2% ,87.8% respectively in cases, and 14.3% ,85.7% respectively in controls, had also no statistical difference between two groups. Interestingly, a significant association is disclosed between the promoter variant - 336G allele and fever in patients ( P = 0. 037, OR = 0. 191, 95 % CI:0. 040-0. 907 ). Conclusion The single nucleotide polymorphism of -871A/G and -336A/G in DCSIGN gene promoter might not be associated with the susceptibility to tuberculosis in Chinese. Tuberculosis patients with -336G allele are significantly protected fever.