ABSTRACT
Objective To explore the application value of T-spot test of Mycobacterium tuberculosis infection (T-SPOT.TB) on diagnosis and differential diagnosis of pulmonary tuberculosis.Methods From Apr.2014 to Dec.2016,700 patients with suspected pulmonary tuberculosis were collected,venous blood (5ml) was drawn off and sputum was collected from each patient separately for T-SPOT.TB and pathogens identification (including TB).Chest CT,bronchoscopy brush or biopsy histopathological examination were followed up,cultivation of My.tuberculosis and of common bacteria with sputum or lavage fluid when needed.T-SPOT.TB test was performed according to the kit instruction operation.2.5 × 105 peripheral blood mononuclear cells (PBMCs) were added into the pre-coated anti-human γ-interferon antibody,and co-incubated separately with two specific My.tuberculosis antigens,namely early secretory targeting 6 (ESAT-6) and culture filtration protein 10 (CFP-10),and then the spot forming cells (SFCs) were counted.The gold standard for present study were set as follows:1) My.tuberculosis smear positive or culture positive;2) Clinical diagnosis (meet any one is positive).The efficacy of T-SPOT.TB on diagnosing active TB was observed,and then the optimal critical value for diagnosing active TB was determined.Patients diagnosed as active TB were divided into 4 subgroups:initial treatment group,retreatment group,smear or culture positive group,and smear or culture negative group.T-SPOT.TB was carried out to detect A and B antigen,and the difference of formed SFCs was then compared.The present study was approved by the Ethics Committee of Xinjiang Uygur Autonomous Region Chest Hospital.Results Of 700 cases suspected of pulmonary tuberculosis enrolled in present study,528 out of 624 definite cases (84.6％) were finally diagnosed as active tuberculosis (active TB group) and 96 cases (15.4％) were as without TB infection (non-TB group).Positive results of T-SPOT.TB test were found in 414 cases in active TB group,and 47 cases in non-TB group were reported with T-SPOT.TB negative.The sensitivity and specificity of T-SPOT.TB test for diagnosing active TB were 78.4％ and 49％,respectively.The positive predictive value,negative predictive value,positive likelihood ratio and negative likelihood ratio were 89.4％,29.2％,1.537 and 0.441,respectively.ROC curve showed that the specificity increased significantly (from 49％ to 62.5％) while the sensitivity decreased (from 78.4％ to 72.7％) when antigen A (cut-off:16.0 SFCs/2.5 × 105 PBMC) was combined with antigen B (cut-off:7.0 SFCs/2.5 × 105 PBMC) for analysis.In addition,the number of A and B antigen spots in active TB group was significantly higher than that in non-TB group (P＜0.01).The number of B antigen spots in positive TB group was significantly higher than that in negative TB group (P＜0.05).There was no significant difference among the other groups.Conclusions Since the high sensitivity and low specificity of T-SPOT.TB in the diagnosis of active TB,the final diagnosis should be combined with clinical manifestations.When the A antigen is 16.0 SFCs/2.5 × 105 PBMC and the B antigen is 7.0 SFCs/2.5 × 105 PBMC,the specificity of T-SPOT.TB will be improved.Higher number of spots has a certain reference diagnostic value for active TB.
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Objective To investigate the effect of evodiamine on proliferation of HCT-116 in balb/c nude mice, and to explore possible mechanism. Methods HCT-116 cells were injected into the right armpit of 4 week old Balb/c nude mice, the feeding had been executed at the time of the diameter of the xenografted tumor reached 0.5 cm,at the dose of 3 mg/kg (Evo), body weight and tumor diameter had been tested every three days, and made the curve of body weight and tumor diameter of the mice. All the mice were sacrificed after 22 days of feed-ing,and harvested the xenografted tumors. The morphological difference of the two groups tumor were identified by HE staining,the expression of HDAC3, NF-κB and p53 protein were detected by IHC and Western blot. Results Compared with the control groups, the volume and weight of the tumors in Evo groups were significantly lighter, and the body weight of the nude mice were heavier,the tumor cells in Evo groups were shrink,deeply staining nu-celus and their abnormal mitoses were fewer,the expression of NF-κB and p53 were increased but HDAC3 was de-creased in xenografted tumors treated with Evo(P<0.05). Conclusions Evo can change the expression of NF-κB and p53 by down-regulating HDAC3,and inhibit the proliferation of HCT-116 cells line in vivo.
ABSTRACT
Objective To investigate the effect of evodiamine on proliferation of HCT-116 in balb/c nude mice, and to explore possible mechanism. Methods HCT-116 cells were injected into the right armpit of 4 week old Balb/c nude mice, the feeding had been executed at the time of the diameter of the xenografted tumor reached 0.5 cm,at the dose of 3 mg/kg (Evo), body weight and tumor diameter had been tested every three days, and made the curve of body weight and tumor diameter of the mice. All the mice were sacrificed after 22 days of feed-ing,and harvested the xenografted tumors. The morphological difference of the two groups tumor were identified by HE staining,the expression of HDAC3, NF-κB and p53 protein were detected by IHC and Western blot. Results Compared with the control groups, the volume and weight of the tumors in Evo groups were significantly lighter, and the body weight of the nude mice were heavier,the tumor cells in Evo groups were shrink,deeply staining nu-celus and their abnormal mitoses were fewer,the expression of NF-κB and p53 were increased but HDAC3 was de-creased in xenografted tumors treated with Evo(P<0.05). Conclusions Evo can change the expression of NF-κB and p53 by down-regulating HDAC3,and inhibit the proliferation of HCT-116 cells line in vivo.
ABSTRACT
Aim To investigate the effect of miR-320a up-regulation on the apoptosis and migration of Bel7402 cells induced by ribonucleic acid Ⅱ.Methods The different expression levels of miR-320a in normal liver cells and hepatocellular carcinoma (HCC) cells were detected by qRT-PCR.Bel-7402 cell was transfected with miR-320a mimic,and the miR-320a expression levels were measured by qRT-PCR.The effect of ribonucleic acid Ⅱ on proliferation of Bel-7402 and Bel-7402-miR-320a cells was measured by CCK-8 assay,and cell cycle and apoptosis were detected by flow cytometry.The migration and invasion ability of ribonucleic acid Ⅱ on Bel-7402 cells were tested by Transwell method.The expression of p53,Cyclin D1,Bax,Bcl-2,MMP-3 proteins were examined by Western blot.Results miR-320a expression levels in HCC cell line Bel-7402 were significantly lower than those in normal cell line HL-7702.Bel-7402 cells were successfully transfected with miR-320a mimic,named Bel-7402-miR-320a.CCK-8 showed that ribonucleic acid Ⅱ could effectively inhibit the proliferation of Bel7402 and Bel-7402-miR-320a cells in vitro in a dosedependent manner at the range of 100,200,300,400,500 mg · L-1.The IC50 of ribonucleic acid Ⅱexposure on Bel-7402 and Bel-7402-miR-320a cells for 12 h and 24 h was 250,200 mg · L-t and 150,120 mg · L-1,respectively;flow cytometric analysis indicated that over-expression of miR-320a could arrest Bel-7402 and Bel-7402-miR-320a cells induced by ribonucleic acid Ⅱ in G0/G1 phase,and promote the apoptosis of HCC cells.Transwell method showed that Bel-7402-miR-320a + Ribonucleic acid Ⅱ group could significantly inhibit the migration of HCC cells compared with control group and Bel-7402 + Ribonucleic acid Ⅱ group.Western blot results showed that the expression of p53,Bax proteins increased,while the Cyclin D1,Bcl-2,MMP-3 proteins were down-regulated in Bel-7402 and Bel-7402-miR-320a cells induced by ribonucleic acid Ⅱ.Conclusions The expression of miR-320a is lower in HCC cells than that in normal cell line.While ribonucleic acid Ⅱ could promote the apoptosis of liver cancer cells by arresting the cell cycle protein expression of Cyclin D1,activating p53 signaling pathway,down-regulating Bcl-2,up-regulating Bax and destroying Bcl-2/Bax proportions,and inhibiting the migration and invasion of HCC cells by downregulating MMP-3.Overexpression of miR-320a could increase the sensitivity and boost the pharmacological effects of ribonucleic acid Ⅱ on HCC cells.