Objecflve To evaluate the performance of two rapid and low-cost metheds(MTT test,and rosazurin mierotitre assay)for the detection of resistance to first-line drugs in Mycobacterium tuberculosis.Methods sixty-four Myeobaeterium tuberculosis clinical isohtes were tested by the MTT test and the rosazuxin microtitre assay(REMA)respectively,and the results were compared with those obtained with the absolute concentration method on L(o)wenstein Jensen medium.Results The MTT test and the resazurin microtitre assay showed a good agreement compared with the absolute concentration method for all first-line drugs tested.The sensitibity,specificity and accuracy of the MTT test were 94.8%,96.0%,95.3%,for RFP;93.8%,93.8%,93.8% for INH;92.9%,96.O%,95.3% for EMB,90.6%,87.5%,89.1% for SM,respectively.The sensitivity,specificity and accuracy of the resazurin microtitre assay were 92.3%,96.0%,93.8%,for RFP;90.6%,90.6%,90.6% for INH;92.9%,94.0%,93.8% for EMB,87.5%,87.5%,87.5% for SM,respectively.The Kappa value of the MTT test and the absolute concentration method for the detection of resistance to RFP,INH,EMB,SM were 0.857,0.831,0.714,0.792.respeedvely;The Kappa value of the regazurin mierotitre assay and the absolute concentration method for the detection of resistance to RFP,INH,EMB,SM were 0.871,0.826,0.826,0.750,respectively.The Kappa value of the MTT test and the resazurin microtitre assay for the detection of resistance to RFP,INH,EMB,SM wefe 0.889,0.875.0.787,0.844,respectively.Conclusions Both MTT test and the resazurin microtitre assays are simple,rapid,low-cost and sensitive for rapid detection of resistance to first-line drugs.They could be promising methods for susceptibility assay of the first-line antituberculosis drugs in low-resource countries.

Objective To explore the value of IFN-γ produced or secreted by CD+4 T Lymphocytes from pleural effusion mononuclear cells for the diagnosis of tuberculous pleurisy(plTB).Methods The PEMCs of 40 patients with tuberculous pleural effusion and 30 patients with malignancy pleural effusion were selected as the tuberculosis and disease control groups, then co-cultured with the early secretory antigenic target 6 (ESAT-6) and culture filtered protein 10 (CFP-10) fusion protein (E/C).The numbers of spot forming cells(SFC) secreting IFN-γ were enumerated by ELISpot and the ratios of cells producing IFN-γ were detected by flow cytometry and intracellular cytokine staining.Moreover, the two indicators were compared between tuberculosis and disease control groups to evaluate the 2 methods detecting IFN-γ in the diagnosis of plTB.Results After E/C stimulation, the numbers of SFC were 205(125-450)SFC/5×104 PEMC in tuberculosis group and 5(2-18)SFC/5×104 PEMC in disease control group by ELISpot.The difference between two groups was statistically significant (U= 20.00, P<0.01).The proportion of IFN-γ-secreting CD+4 T lymphocytes was 3.27% (1.81%-7.34%) in tuberculosis group and 0.12% (0.06%-0.46%) in control group detected by FCM. The difference between the two groups was statistically significant (U=45.00, P<0.01).The indicators of ELISpot in detection of IFN-γ which was secreted by PEMC after co-cultured with E/C were as follows: sensitivity 92.5% (37/40), specificity 80.0% (24/30), positive predictive value 0.86, negative predictive value 0.89, positive likelihood ratio 4.63, negative likelihood ratio 0.09 and accuracy 87.1%;and for FCM, they were 87.5% (35/40), 90.0% (27/30), 0.92, 0.84, 8.75 and 0.14, respectively and accuracy 88.6%.Conclusion After E/C stimulation, the assay for IFN-γ-secreting CD+4 T lymphocytes by FCM and ELISpot is highly sensitive and specific for diagnosis of plTB as an auxiliary method.

Objective To establish the fingerprint analysis method of Schisandrae Chinensis Fructus by HPLC; To analyze the similarity on the fingerprints of Schisandrae Chinensis Fructus from different producing areas. Methods The chromatographic separation was performed by HPLC on a Diamonsil C18 column (250 mm×4.6 mm, 5 μm). Acetonitrile-water was used as gradient mobile phase. The flow rate was 1.0 mL/min. The column temperature was maintained at 30℃. The detection wavelength was set at 254 nm.Results The HPLC fingerprint analysis method of Schisandrae Chinensis Fructus was established. Twenty-nine common fingerprint peaks were identified. The similarities of the fingerprints of ten samples from different producing areas were above 0.95.Conclusion The method is simple and reliable, which can provide a scientific basis for quality evaluation of Schisandrae Chinensis Fructus.

stinguish active tuberculosis and healthy cases with tuberculosis exposure history according SFC count.

OBJECTIVE:To study the improvement effect of Celosia cristata n-butanol extracts on dysfunctional uterine bleed-ing of rats,and explore its mechanism. METHODS:60 pregnant SD rats were randomly divided into blank group,model group, Gongxuening capsule group (positive control,0.07 g/kg) and C. cristata n-butanol extracts high-dose,medium-dose,low-dose groups(4.32,2.16,1.08 g/kg),10 in each group. Except for the blank group,rats in other groups were intragastrically given mife-pristone and misoprostol on 7th of pregnancy for resulting incomplete abortion to induce models of dysfunctional uterine bleeding. Then rats in administration groups were intragastrically given relevant medicines,rats in blank group and model group were intra-gastrically given normal saline once every morning and evening,for 7 d. On 8th d of pregnancy,uterine bleeding amount,and thromboxane (TXA2),prostacyclin (PGI2) and tumor necrosis factor α(TNF-α) contents in serum were determined. RESULTS:Compared with blank group,uterine bleeding amount in model group was significantly increased(P<0.01),TXA2 content in se-rum was significantly reduced,PGI2 and TNF-α contents were significantly increased(P<0.01). Compared with model group,uter-ine bleeding amounts in administration groups were significantly reduced,TXA2 content in serum was significantly increased(P<0.01);PGI2 and TNF-α contents in serum in Gongxuening capsule group and C. cristata n-butanol extracts high-dose group and TNF-α content in serum in C. cristata n-butanol extracts medium-dose group were significantly reduced (P<0.01). CONCLU-SIONS:C. cristata n-butanol extracts show obvious improvement effect on incomplete drug abortion-induced dysfunctional uterine bleeding of rats,and the mechanism may be related to the regulation of TXA2/PGI2 dynamic balance and inhibition of TNF-α tran-sient secretion.

BACKGROUNDTo evaluate the values of a new tumor marker carbohydrate antigen (CA242) and combined determination of CA242, tissue polypeptide antigen (TPA), neuron-specific enolase (NSE) and carcinoembryonic antigen (CEA) in the diagnosis of malignant pleural effusion associated with lung cancer.

METHODSThe concentration of CA242, TPA, NSE and CEA in the serum and the pleural effusion was measured in 57 patients with malignant pleural effusion associated with primary lung cancer and 30 patients with tuberculous pleural effusion by enzyme-linked immunosorbent assay.

RESULTSThe levels of the four tumor markers in the serum and pleural effusion from patients with lung cancer were significantly higher than those with tuberculous pleural effusion (P < 0.01). The sensitivity of CA242 in the serum and the pleural effusion for lung cancer was 53.6% (31/57) and 61.4% (35/57) respectively; the sensitivity of CA242 for lung adenocarcinoma was 65.7% (23/36) and 66.7% (24/36) respectively. The specificity was 90.0%. Combined determination of the four tumor markers in serum and pleural effusion: If two or more of them were positive for evidence for diagnosis of lung cancer, the specificity for the serum and the pleural effusion was 96.7% (29/30) and 100.0% (30/30) respectively, with the sensitivity of 75.4% (43/57) and 77.2% (44/57) respectively.

CONCLUSIONSThe determination of the new tumor marker CA242 in serum and pleural effusion might be useful for the diagnosis of malignant pleural effusion associated with lung cancer, especially for adenocarcinoma. The combined determination of the four tumor markers can increase the specificity and the sensitivity in the diagnosis of malignant pleural effusion.

BACKGROUND AND OBJECTIVEPromoter hypermethylation of the RASSF1A gene is among the most abundant epigenetic deregulations in human cancer. The aim of this study is to investigate the relationship between the methylation status of RASSF1A promoter and the prognoses of non-small cell lung cancer (NSCLC).

METHODSThe methylation status of RASSF1A promoter in 150 NSCLC and 25 non-malignant tissues was determined using a methylation-specific polymerase chain reaction (MSP).

RESULTSRASSF1A promoter hypermethylation was detected in 38.7% (58/150) of NSCLC tissues, but in none of the non-malignant tissues. The patients with hypermethylation of RASSF1A had a poor survival rate, and the relationship between the survival rate and hypermethylation of RASSF1A was statistically significant (P = 0.004). Then by using stepwise Cox proportional hazard regression testing, methylation status of RASSF1A was an independent factor affecting the NSCLC patients' survival (RR = 1.584, 95% CI: 1.040-2.411, P = 0.032).

CONCLUSIONThe hypermethylation of the RASSF1A promoter may be an independent prognostic factor of NSCLC after operation.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; genetics ; pathology ; DNA Methylation ; genetics ; Female ; Humans ; Lung Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Prognosis ; Promoter Regions, Genetic ; genetics ; Tumor Suppressor Proteins ; genetics