Cerebrospinal fluid (CSF) and serum from patients with tuberculosis meningitis (TBM) and nontuberculosis meningitis were assayed for IgG antibody activity to purified protein derivative (PPD) by ELISA.The results showed that the patients with TBM clearly had higher levels of antibody to PPD antigen than did approrjate control groups (P0.05).Because of its sensitity, specificity,and rapidity value,this method can be used for early diagnosis of tuberculosis meningitis.

Objective To study the influencing factors of pediatric in-hospital cardiopulmonary resuscitation(CPR).Methods It was a retrospective observational study.We studied a total of 281 children who suffered in-hospital cardiopulmonary arrest(CPA).The outcome was defined as sustained return of spon-taneous circulation ﹥20 min.Results A total of 281 patients met study entry criteria.In 129 children (45.9%),return of spontaneous circulation sustained ﹥20 min and 20 cases(8.8%)survived to hospital discharge.In the univariate analysis,gender,age,weight,time of CPA happened,first cardiac rhythm,pH, blood lactate had no obvious influence on the outcome.Underlying disease,the place of CPA,personnel fac-tors,airway support,the duration of CPR,doses of adrenaline,use of bicarbonate and blood glucose level were associated with outcome.Conclusion At present,the rate of successful CPR and discharge of hospital is still low.Respiratory disease has a higher survival rate.CPR ﹥20 min,use of bicarbonate and using adrenaline≥3 doses are associated with poor outcome and an increase in mortality.

Objective To develop a new Multiplex Allele-Specific polymerase chain reaction(MAS-PCR) assay to detect the main point mutations in the katG and rpoB genes, which has been reported to account for the majority of clinical Mycobaterium tuberculosis resistant to isoniazid and rifampin. Methods Based on the sequences of katG and rpoB genes, specific primers were designed to carry out the MAS-PCR to detect the most common mutations in codon315 of katG and codons 531,526,516 of rpoB gene. Results The purified DNA preparation of 96 clinical Mycobacterium tuberculosis strains were used to optimize PCR. No mutation was detected in 19 isoniazid-sensitive strains. The 315Ser point mutation was detected in 79.2%(61/77)of isoniazid-resistant strains, the type of mutation includes the most common S315T and the less common S315N, which could’t be detected by PCR-RFLP(polymerase chain reaction-restriction fragment length polymorphism). However, S315G couldn’t be detected by MAS-PCR and that will make a false negative. The mutations in codons 531,526,516 were detected by the MAS-PCR. Compared with the results of direct sequencing of rpoB gene, no mutation was detected in sensitive strains. For rifampin-resistant strains, the total sensitivity was 81.5%(66/81). Conclusions MAS-PCR is a new molecular method with high sensitivity and specificity, which can be used to detect the point mutation in katG and rpoB gene rapidly and economically. It can be used in clinical laboratories to detect drug-resistant tuberculosis strains. Simultaneous detection for katG and rpoB gene mutations in one MAS-PCR system will help to improve the efficiency of this method.

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 investigate the correlation of FAT10 expression with the malignant characteristics of hepatocellular car-cinoma (HCC), and to explore the effect of FAT10 on RhoA and cytoskeleton of HCC. Methods: Immunohistochemistry (IHC) was used to detect the FAT10 expression level of 108 HCC patients, and the correlation between the expression of FAT10 and the malignant characteristics of HCC patients was analyzed. We transiently transfected plasmids with overexpressed FAT10 using 7721 and HepG2 cells or interfered with FAT10 expression using siRNA in Huh7 and LM3 cells. Active-RhoA, total-RhoA, and ROCK protein expres-sion levels were detected by Western blot analysis after overexpression or interference. We also used immunofluorescence to detect changes in the cytoskeleton protein F-actin after FAT10 overexpression in 7721 cells. Results:Correlation analysis showed that both ac-tive-RhoA and FAT10 expression levels were significantly correlated with clinical malignant characteristics by using IHC (RhoA:me-tastasis, P=0.036 and recurrence, P=0.026;FAT10:metastasis, P=0.031 and recurrence P=0.004). In addition, active-RhoA expression level was correlated with FAT10 (P=0.000). Survival analysis showed that the prognoses of low-expression active RhoA (P=0.019) or FAT10 (P=0.026) groups were significantly better than those of the high-expression groups. Western blot analysis showed that FAT10 increased the expression of active-RhoA and ROCK. However, the expression of active-RhoA and ROCK decreased after FAT10 inter-ference. F-actin expression increased in the 7721 cells with overexpressed FAT10 (all P<0.01). Moreover, FAT10 facilitated F-actin ag-gregation on cell membrane and changes in F-actin. Conclusion:FAT10 is correlated with the malignant characteristics of HCC and may promote changes in HCC cytoskeleton induced by active-RhoA.

Objective To study the role of extra-anatomic arterial bypass grafting in the management of aorta-iliac occlusive disease. Methods Forty-one cases of aorta-iliac occlusive disease managed by extra-anatomic arterial bypass grafting were retrospectively analyzed. Clinical features included claudicafion 11, resting pain 12 and ulcer/gangrene 18 patients. Axillofemoral bypass was performed in 15 cases with aortic and bilateral iliac artery occlusion, femorofemoral bypass in 18 cases with unilateral iliac occlusion and femoro-cnntralateral popliteal bypass in 8 cases with iliac and ipsilateral femoral artery occlusion. Results Postoperatively, clinical features were reheved in 40 cases (97.6%). Ankle brachial index increased significandy from 0.41 ± 0.15 to 0.89 ± 0.21 perioperatively (P < 0.05 ).Thirty-five cases were followed up for 1 year to 5 years. The overall graft patency rates in this series were 93%, 86% and 80% in 1, 3 and 5 years, respectively. Conclusion Extra-anatomic arterial bypass has merits as less invasive and rapid postoperative recovery, and this is one of the effective methods for the treatment of aorta-iliac occlusive disease.

Objective To evaluate the accuracy of pyrosequencing for the mutation detection of katG gene in isoniazid resistance in Mycobacterium tuberculosis using Meta-analysis.Methods Searching PubMed,Web of Science,Elsevier,and China National Knowledge Infrastructure (CNKI),Weipu and WANFANG DATA to obtain the relevant English and Chinese-language articles,respectively.A written protocol and explicit study selection criteria was followed.Quality of included trials was assessed by QUADAS (quality assessment of diagnostic accuracy studies).Subsequently,the characteristics of the included articles were appraised and extracted.Heterogeneity of the included articles was tested by using STATA 10.0,which was used to select proper effect model.The fixed effects model was adopted using Meta-Disc software.Finally,sensitivity analysis was performed.Results Totally 114 research papers were collected and 9 articles were selected.The accordance between pyrosequencing and conventional sequencing result was 100％.Eight studies were involved including 945 specimens when katG gene was detected.The overall sensitivity and specificity were 0.77 (0.73,0.80) and 1.00 (0.99,1.00),respectively.The area under the SROC was 0.9882.As inhA gene was detected,the overall sensitivity and specificity were 0.19 (0.15,0.24) and 1.00 (0.98,1.00).The test was stable.Conclusions Our meta-analysis reveals that pyrosequencing is a highly specific tool for detection mutation of katG gene of isoniazid resistance.This result suggests that it is useful for screening of isoniazid resistance in diagnostic test.(Chin J Lab Med,2013,36:329-332)

Objective To develop new method for rapid detection of mutations in rpoB gene related with resistance to rifampin of M. tuberculosis . Methods According to the sequence of wild type M. tuberculosis, five oligonucleotide probes covering the 69 bp hyper variable region of rpoB gene were designed and immobilized on nylon membrane strips. Thereafter, the target rpoB gene fragment was obtained by PCR using biotin labeled primers and thereafter the PCR product was denatured and hybridized with probes on membrane. The results of reverse dot blot hybridization were compared with the results of drug sensitivity test and sequencing. Results PCR products from 36 RFP resistant and 22 RFP susceptible isolates were detected by the assay of reverse dot blot hybridization, showing that the susceptibility and the specificity rate were 88.9% and 86.4% respectively,and the coincidence with biochemical method and sequencing is 87.9% and 89.7% respectively. Conclusions Reverse dot blot hybridization is a rapid and sensitive method to detect the rpoB gene mutations,which may be used in early detection of the resistance of M. tuberculosis to rifampin.

Objective To elucidate the relationship between polymorphisms of NRAMP1 gene and susceptibility to tuberculosis in Chinese Han nation by case control study. Methods We selected 127 patients with active pulmonary tuberculosis who were all Han people with mean age of 52.5 years and 58 ethnically matched healthy controls. We typed the polymorphisms of NRAMP1, INT4 and D543N, through polymerase chain reaction restriction fragment length polymorphism (PCR RFLP) techniques. Results Two NRAMP1 polymorphisms were both significantly associated with smear positive pulmonary tuberculosis. Subjects who were heterozygous for polymorphisms in INT4 and D543N were related with tuberculosis. It was the cooperation of different polymorphisms of NRAMP1 that led to the susceptibility to tuberculosis. Conclusions The polymorphisms in NRAMP1 gene affect susceptibility to tuberculosis in Han people in China.