Objective To investigate the isoniazid-dependence of clinical isolated strains of M.tuberculosis.Methods The double-phase Kuang's agar medium was employed to isolate M.tuberculosis from the patient's sputum,and its drug-resistance and drug-dependence were determined.Results 184 clinical isolates of isoniazid-dependent M.tuberculosis were grouped as follows.Type A:the strains grew faster and more vigorous in drug tubes than in control tubes,and there were 12 strains(6.5%)which grew more colonies in high concentration tubes than in low concentration tubes;Type B:the strains grew faster and more vigorous in drug tubes than in control tubes,and there were 10 strains(5.4%)which grew more colonies in low concentration tubes than in high concentration tubes;Type C:the strains grew faster and more vigorous in low concentration tubes than in control tubes,and there were 15 strains(8.15%)which grew less colonies in high concentration tubes than in control tubes;Type D:the strains grew faster and more vigorous in high concentration tubes than in control tubes,and there were 4 strains(2.2%)which grew less colonies in low concentration tubes than in control tubes.The isoniazid-dependent strains covered 22.3%(41/184)of the total clinical isolates.Conclusion The isoniazid-dependent strains did exist in the clinical isolates of M.tuberculosis,and different features of dependence have been detected.

Objective To investigate the feasibility of multiplex PCR for rapid identification of Mycobacterium tuberculosis and non-tuberculosis Mycobacteria. Methods According to MTP40 gene sequence of Mycobacterium tuberculosis, 32kD gene sequence of Mycobacterium and IS6110 insertion sequence gene sequence of Mycobacterium tuberculosis complex, three specific pairs of primers (PT1-PT2, MT1-MT2 and IS5-IS6) for Mycobacterium were designed, and the target DNA for MTP40, 32kD and IS6110 was 396bp, 506bp and 984bp, respectively. The genome of 92 Mycobacterium tuberculosis clinically isolated strains and 5 non-tuberculosis Mycobacteria clinical strains were amplified in the same system, and the results were compared with reference strains. Results Among 92 clinical strains of Mycobacterium tuberculosis, the DNA fragments of 396bp, 506bp and 984bp were found in 90 Mycobacterium tuberculosis clinical strains, as well as in the reference strain H37Rv; the sensitivity of multiplex PCR for Mycobacterium tuberculosis was 97.8%, and the specificity was 100.0%. The DNA fragments of 506bp were all found in 5 non-tuberculosis Mycobacteria clinical strains, the sensitivity and specificity for non-tuberculosis Mycobacterium were both 100.0%. Conclusion The multiplex PCR is a rapid, sensitive and specific method for identification of Mycobacterium tuberculosis and non-tuberculosis Mycobacteria, and it may provide an effective way for clinical diagnosis of Mycobacterium tuberculosis and non-tuberculosis Mycobacteria, therefore useful in clinical application.

Objective To investigate whether emodin(1,3,8-trihydroxy-6-methylanthraquinone) can attenuate inflammatory response in rats' lungs with acute necrotic pancreatitis (ANP). Method Acute necrotic pancreatitis model was induced by injection of 3% sodium taurocholic acid into the subcapsular of pancreas and emodin was administered by intestine perfusion. Plasma amylase of 3, 6 and 12 h with acute necrotic pancreatitis was measured together with the detection of IL-1β, IL-6 and IL-10 mRNA expressions in rats' lungs by semi-quantitative RT-PCR. Immunohistochemistry was detected the expression of IL-1β converting enzyme (ICE) in the rats' lungs. Result Plasma amylase of 3, 6 and 12 h with acute necrotic pancreatitis groups are obviously high as compared with normal group(P＜0.05). Plasma amylase was (1 611.20±218.72)IU/L in normal group. Plasma amylase of 3, 6 and 12 h with acute necrotic pancreatitis groups were (1 981.40±56.81)IU/L, (3 287.40±612.37)IU/L and (4 914.60±746.82)IU/L. Plasma amylase of 3, 6 and 12 h with acute necrotic pancreatitis after treatment with emodin groups were obviously low as compared with acute necrotic pancreatitis groups. The plasma amylase was(1 617.20±136.80)IU/L,(2 323.40±318.19) IU/L and (2 670.20±390.03)IU/L respectively. The study showed that the mRNA expression of pro-inflammatory cytokin IL-1β and the expression of IL-6, as well as the expression of IL-1β converting enzyme(ICE) were decreased and IL-10 was increased. Conclusion The study demonstrates that emodin plays an important role in reducing plasma amylase level. Emodin exerts anti-inflammatory effects in acute necrotic pancreatitis rats' lungs by downregulating the mRNA expression of IL-1β and IL-6 and upregulating the mRNA expression of IL-10.

Objective To analyze the dielectrophoresis patterns of the proteome of the Rifampin-dependent and-resistant stains of Mycobacterium tuberculosis,to search and identify the differently expressed proteins,and to provide the proteomic basis for researching the mechanism of anti-tuberculosis drug dependence of M.tuberculosis.Methods The whole somatic proteins were extracted from two strains of M.tuberculosis.The first dimensional ampholine electrophoresis was performed on immobilized pH gradient(IPG) rod gels(pH 4-7).Then the proteins on IPG strips were separated using SDS-PAGE.The stained gels were scanned with image scanner and the images were analyzed by Imagemaster 2D software.The differentially expressed proteins were detected by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS).Results Seven hundred and fifty-three spots were detected in Rifampin-dependent strain of M.tuberculosis,while 584 spots were detected in Rifampin-resistant strain,including 404 match spots(the match rate: 61.5%).As to the expression in Rifampin-dependent strain,7 spots significantly up-regulated and 35 spots down-regulated,6 spots were absent in expression,and 5 spots expressed separately,most of the spots were small molecular proteins.Ten spots were selected to run MS analysis.Nine spots were identified as representing 7 proteins.Conclusion The Rifampin-dependent strain of M.tuberculosis is characterized by a rapid and vigorous growth mainly by means of the differential expression of enzymes related to energy metabolism and fatty acid biosynthesis.

Objective To observe the correlation between CEUS quantitative parameters of carotid plaques and leukocytes in patients with acute ischemic stroke.Methods Sixty-two patients with large artery atherosclerosis stroke (LAAS group) confirmed by CT or MRI were enrolled,while 54 patients in the same period of hospitalization,age and gender-matched,no history of cardiovascular events with atherosclerosis were taken as control group.The correlation between CEUS quantitative parameters of carotid plaques and leukocytes in two groups were compared.Multiple linear regression model was built and the risk factors of CEUS quantitative parameters were analyzed.Results The total leukocytes count,neutrophils count and neutrophil/lymphocyte ratio in LAAS group were higher,while the lymphocytes count was lower than those in control group (all P＜0.05).CEUS parameters,including timeqntensity curve (TIC) peak (TIC-P),mean (TIC-M),fitting curve (FC) peak (FC-P),sharpness (FC-S) and area under the curve (FC-AUC) of carotid plaques were higher than those in control group (all P＜0.05),while neutrophils count and neutrophil/lymphocyte ratio were positively correlated with FC-AUC (r=0.298 and 0.739,respectively;all P＜0.05).Total leukocytes count was independent risk factor of TIC-P,and neutrophil/lymphocyte ratio was independent risk factor of FC-AUC (all P＜0.05).Conclusion CEUS quantitative parameters of carotid plaques related to leukocytes count.Increased leukocytes or neutrophil/ lymphocyte ratio might rise vulnerability of plaques.