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Objective:To investigate the changes of T-lymphocyte subsets, T-cell immunoglobulin and mucin domain molecule-1 (TIM-1) and TIM-3 levels, and cytokines in the peripheral blood of patients with active tuberculosis.Methods:From December 2017 to December 2018, 50 tuberculosis patients and 50 cured tuberculosis patients in Zhejiang Hospital of Integrated Chinese and Western Medicine were selected as the tuberculosis group and cured tuberculosis group, respectively. Fifty healthy individuals in the same period were selected as the control group. Flow cytometry was used to detect the T-lymphocyte subsets in the peripheral blood. The mRNA levels of TIM-1, TIM-3, interferon(IFN)-γ and interleukin(IL)-4 in peripheral blood mononuclear cells (PBMC) were detected by quantitative real-time polymerase chain reacticn (PCR). T test was used for statistical analysis. Results:The ratio of CD4 + /CD8 + T lymphocytes in the tuberculosis group (1.21±0.50) decreased significantly, comparing with those in the cured tuberculosis group (1.88±0.62) and the control group (1.92±0.82). The differences were statistically significant ( t=2.148 and 2.207, respectively, both P<0.05). The mRNA levels of TIM-1, TIM-3 and IL-4 in PBMC in the tuberculosis group were 2.16±0.37, 1.59±0.36 and 1.52±0.69, respectively, which were all higher than those in the cured tuberculosis group (1.60±1.23, 1.01±0.52 and 0.91±0.36, respectively) and the healthy control group (1.40±0.27, 0.92±0.34 and 0.79±0.42, respectively). All of these differences were statistically significant ( t=14.120, 11.440, 17.130, 12.090, 12.050 and 17.030, respectively, all P<0.05). However, the IFN-γ mRNA level (0.43±0.11) was lower than that in the cured tuberculosis group (1.74±0.72) and the control group (1.82±1.17), and the differences were both statistically significant ( t=13.880 and 11.430, respectively, both P<0.05). Conclusion:The immune dysfunction in patients with active tuberculosis may be related to the low ratio of CD4 + /CD8 + T lymphocytes, the increased expressions of TIM-1 and TIM-3, and the imbalance of helper T lymphocyte (Th)1/Th2 cytokines.
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Objective To analyze the expression of miRNA-29a in U937 macrophages infected with Mycobacterium tuberculosis and its regulation of target genes.Methods The target genes of miRNA-29a were predicted with softwares miRnada,RNAhybrid and targetscan.The differentiation of U937 macrophages was induced by phorbol ester(PMA), the induced U937 cells were infected with bacilli calmette guerin (BCG).The expression levels of miRNA-29a and its target genes in U937 cells were detected with real-time fluorescence quantitative PCR(RT-fqPCR).The miRNA-29a over-and low-expression U937 macrophage cell lines were constructed and the levels of miRNA-29a and its garget genes were detected.The SPSS 18.0 software was used to analyze the data.Results As predicted by relevant softwares,the miRNA-29a regulate the expression of VEGFA,NFATC3 and TSC22D3 genes.After BCG infection,the expression of miRNA-29a increased to 1.33 fold(P <0.05), and the expression levels of VEGFA, NFATC3 and TSC22D3 were increased to 5.34,99.25 and 2.12 fold,respectively(P<0.01).In the miRNA-29a over-expressing U937 macrophages,the expression levels of VEGFA,NFATC3 and TSC22D3 were up-regulated to 1.35,1.29 and 3.38 fold,respectively(P<0.05 or <0.01).While in the U937 macrophages with miRNA-29a knock-down,the expression levels of VEGFA, NFATC3 and TSC22D3 were down-regulated to 0.07, 0.08 and 0.55 fold, respectively(P <0.01).Conclusion The results suggest that Mycobacterium tuberculosis infection can increase the expression of miRNA-29a in U937 macrophages,further targeting the regulation of VEGFA, NFATC3 and TSC22D3 gene expression, which may participate in the pathogenesis and development of tuberculosis.
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Objective To investigate the effects of human dendritic cells ( DCs) infected by bovine Mycobacterium tuberculosis attenuated live bacteria ( BCG) on differentiation of CD4 +naive T cells from neonate cord blood .Methods After infected with BCG , human DCs were cultured with CD 4 +naive T cells from neonate cord blood, the expression of miRNA-99b in DCs and the expression of Foxp3, ROR-γt, IFN-γand IL-10 mRNAs in CD4+ T cells were detected by qRT-PCR.DCs were transfected with miRNA-99b antisense oligonucleotides and co-cultured with neonatal cord blood CD 4 +naive T cells , and the transcription level of CD4 +T cell-related genes was detected .SPSS 15.0 was used to analyze the data .Results The transcriptional activity of miRNA-99b gene in BCG-infected DCs was significantly higher than that in uninfected DCs (t=7.06,P<0.01).Compared with CD4 +T cells co-cultured with uninfected DCs, the mRNA expression of IFN-γ(45.61 ±4.46 vs.3.54 ±1.73, t=32.32, P<0.01), IL-10 (4.17 ±1.06 vs.1.26 ±0.67, t=2.24, P<0.05) in CD4 +T cells co-cultured with BCG-infected DCs was significantly increased, while the mRNA expression of ROR-γt was significantly decreased ( 0.08 ±0.02 vs.0.63 ± 0.10, t=0.42, P<0.01).Compared with CD4 +T cells co-cultured with DCs transfected with NC-siRNA, the miRNA-99b expression was blocked , the mRNA expression of Foxp3 (0.12 ±0.01 vs.1.57 ±0.90, t=1.06, P<0.05), IFN-γ(0.03 ±0.01 vs.0.64 ±0.35, t =0.44, P<0.05), IL-10(0.03 ±0.01 vs. 0.76 ±0.09, t=0.54,P<0.01) in CD4 +T cells was significantly decreased , while the expression ROR-γt mRNA was significantly increased (17.03 ±5.51 vs.1.32 ±0.14, t=11.54,P<0.01).Conclusion BCG induces the imbalance of initial CD 4 +T lymphocytes into Th17/Treg by regulating the expression of miRNA-99b in DCs, leading to the occurrence and development of infection .
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Objective To discuss the impact of repeated contrast media exposure on renal function in patients who received coronary angiography ( CAG) or percutaneous coronary intervention ( PCI) within 1 week after CTA of coronary ateries. Methods A total of 258 patients who received CAG or PCI after coronary CTA were divided into the study group ( n=132, patients had CAG/PCI within 1 week after CTA) and the control group ( n=126, patients had CAG/PCI 1-2 weeks after CTA). Serum creatinine, cystatin C and estimated GFR were tested before and on day 1, 2 and 3 after procedures. The occurance of contrast-induced nephropathy ( CIN ) was recorded. Resu1ts The baseline clinical characteristics of the patients between the two groups had no significant difference. Preoperative and postoperative serum creatinine, cystatin C and eGFR values on day 1, 2 and 3 had no significant difference between the two groups (all P﹥0. 05). There was no significant difference in the incidence of CIN between two groups (5. 3% in the study group vs. 4. 8% in the control group, P﹥0. 05 ) . Conc1usions It is safe and feasible for patients with eGFR≥60 ml/( min?1. 73 m2 ) to undergo CAG or PCI within 1 week after coronary CTA.
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Objective To study the effects of Qingjinkangkuoyin (QJKKY) on TNF-α and NE in rats with bronchiectasis.Methods Models were established by intrabronchially injecting with pseudomonas aeruginosa,and divided into 5 groups by random:the QJKKY high dose treatment group (given high dose of QJKKY into stomach),the QJKKY low dose treatment group (given low dose of QJKKY),the levofloxacin group (given levofloxacin),the model group (given normal saline),and the normal contrast group (given normal saline).After 2 weeks of treatment,the histopathology of lung tissue,the levels of TNF-α and inflammatory cells in peripheral blood and NE in rats' lung tissue were detected.Results Compared with the model group (160.425±9.9293)ng/L,QJKKY could decrease the level of TNF-α in blood significantly [high dose of QJKKY treatment group was (137.133±6.1646)ng/L,P＜0.05]; the expression of inflammatory cells in serum were decreased significantly by QJKKY [high dose of QJKKY treatment group was (1.106± 0.3580) 109/L,P＜0.05].Low dose of QJKKY treatment group was (1.086 ±0.2433) 109/L,(P＜0.05) ; the expression of NE in lung tissue were decreased remarkably by QJKKY [high dose of QJKKY treatment group(80.697 ±4.5877)ng/L,P＜0.05]; low dose of QJKKY treatment group is (80.747±3.6925)ng/L,(P＜0.05); and the histopathologic change of lung tissue in QJKKY treatment groups were ameliorated under light microscope by HE staining.Conclusion Qingjinkangkuoyin could cure bronchiectasis by decreasing the expression of TNF-αin peripheral blood and NE in rats' lung tissue.
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Objective To evaluate the efficacy and safety of combination therapy with domestic bezafibrate and fluvastatin in patients with combined hyperlipidemia. Methods 180 patients with combined hyperlipidemia were randomly divided into two groups. They were assigned to receive 40 mg fluvastatin (n = 90) or a combination of 400 mg bezafibrate and 40 mg fluvastatin (n = 90) for 24 weeks. Results After 24 weeks treatments, the serum TC, LDL-C levels were reduced (P <0.01) and HDL-C level was increased more significantly (P <0.05) in the combi-nation therapy group. Conclusion Combination therapy with bezafibrate (400 mg) and fluvastatin (40 rag) is more effective than fluvastatin(40 mg) monotherapy.