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 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 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 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.

Ablation Techniques ; adverse effects ; Animals ; Bone and Bones ; pathology ; Disease Models, Animal ; Extremities ; Swine

Objective To observe destruction of tibias created in ablating of the swine limbs in vivo with radiofrequency,the influence on the surrounding organization structure,and the repair process of lesions for treating bony tumor.Methods There were eight pigs included in the first part of the experiment.RFA was performed under DSA guidance at the same position of the diaphysis and the upper end of their right tibias in hind leg during anesthesia,then X-ray and CT examinations were performed at different time points after RFA,and subsequently the pigs were executed by depth anesthesia.Control specimens,specimens of the immediate time,the third day,the tenth day,the twenty-fourth day,the fifth week,the eighth week,the twelfth week after RFA were obtained at the diaphysis and the upper end of their tibias.X-ray and CT examinations of these specimens were performed.These specimens were observed both by naked eye and under microscope.The observation included the general state of the laboratory animals after RFA,the configuration of RFA lesions in the gross specimen and the corresponding histologic changes,the boundary of necrosis in RFA lesions and the corresponding boundary of the gross specimen.There were sixteen pigs in the second part of the experiment.RFA was performed at the same position of the diaphysis and the upper end of their right and left tibias in hind leg during anesthesia,and the pigs were immediately executed by depth anesthesia after RFA to get thirty-two capitulum specimens and thirty-two diaphysis specimens.The scope of necrotic tissue in RFA lesions was observed and measured.The border and scope of necrotic tissue in RFA lesions were observed.Major axis,minor axis,and necrosis volume were measured.The independent sample t test was employed to compare the volume of necrotic tissues between capitulum and diaphysis.The one sample t test was employed to analyze volume of necrotic tissue among capitulum,diaphysis and the standard data.Results No accidental death of the animals occured in the first experimental.The loss of neurological function of the hind leg did not happen.Coagulation necrosis in cancellous bone of capitulum and marrow of diaphysis lesions created in ablating was observed.On 3th to 10th day after RFA,inflammatory cells infiltrated in hemorrhage tape and then granulation tissues formed.Granulation tissues replaced necrotic bone gradually from outside to inside.Low density band was showed on images of X-ray and CT examination in the specimen of 10th day,and it extended inward.The immature bone was observed in fibrous tissue sited at the edge of lesions,which appeared as sclerosis on X-ray and CT examination.The scope of necrotic tissue in RFA lesions included coagulation necrosis and hemorrhagic zone of its periphery.The dead bone and secondary pathological fracture appeared in two of eight pigs.RFA hardly damaged compact bone in the diaphysis,but the damage of soft tissue was observed outside the compact bone.In the second experiment,the shape of lesions in cancellous bone of capitulum was ellipsoid,with long axis of(2.59±0.21) cm,minor axis of(2.15±0.10) cm,and volume of(6.34±0.27) cm3.The shape of lesions in the diaphysis of tibias was ellipsoid,with long axis of(4.53±0.20) cm,minor axis of(1.71 ±0.22) cm,and volume of(7.14±0.36) cm3.There was significantly difference of necrosis volume between the capitulum and diaphysis lesions,(t=2.011,P＜0.05).The virtual necrosis volume of the capitulum and diaphysis lesions was significantly different from the presumed volume with RFA instrument(t=-613.371,-295.878,P＜0.01).Conclusion Cancellous bone of the capitulum can be damaged by RFA,but the compact bone of the diaphysis can hardly be damaged.The intact compact bone can be helpful to protect the surrounding soft tissues.X-ray and CT examination can be used in the evaluation of curative effect of RFA for treating bone tumors.Pathological fracture of the diaphysis may appear after RFA.

Objective To evaluate clinical application of phage amplified biologically assay (PhaB) in susceptibility test of Mycobacterium tuberculosis (MTB) in sputum. Methods The drug susceptibility of MTB was detected by PhaB in 143 patients with sputum-positive pulmonary tuberculosis (PTB),and the chemotherapy regimens were adjusted according to the results of susceptibility test.Independent samples t-tests were used for comparison of means.Count numbers were compared with Chisquare test.If there were count number of 0,Fisher probabilities should be used.ResultsThe total positive rate of PhaB was 94.4％ (135/143) with no differences between three types of PTB (x2 =1.886,P ＞ 0.05 ).The duration of testing for PhaB group was (6.6 ± 1.8) days,while for control was (29.4 ±8.7) days (t =29.01,P ＜ 0.01 ).Compared with control group,the 2-month negative-conversion rate (63.2％ vs.35.1％,x2 =3.989,P ＜ 0.05 ) and cure rate ( 100％ vs.78.4％,P ＜ 0.05 ) of PhaB group in type Ⅱ patients were significantly higher.But there were no differences between PhaB and control groups in type Ⅰ and Ⅲ PTB patients.ConclusionThe results of PhaB drug susceptibility test can be helpful for choosing effective chemotherapy regimen for PTB patients rapidly.

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.