1.Effect of rapamycin on the proliferation and apoptosis of glomerular mesangial cells
Bo FU ; Sifang ZHANG ; Li ZHUO ; Guangyan CAI ; Yuansheng XIE ; Ribao WEI ; Qiang MA ; Xiangmei CHEN
Chinese Journal of Nephrology 2009;25(11):849-852
Objective To investigate the effect of different concentrations of rapamycin on the proliferation and apoptosis of glomerular mesangial cells(GMCs)and to investigate the mechanism. Methods GMCs were treated with different concentrations of rapamycin(1 μg/L,2 μg/L,4 μg/L,8 μg/L,16 μg/L).After treatment for 24 h,48 h and 72 h,cell proliferation was assessed bv MTT colorimetric assay and the growth curve was traced.After treatment for 72 h,the cell cycle distribution and the apoptotic rate of GMCs in different concentrations of rapamycin were analyzed bv flow cytometry.The effects of different concentrations of rapamycin on the mRNA and protein expression of p27 and p53 were detected by RT-PCR and Western blot respectivelyResult The low dose of rapamycin(1 μ/L)could signiticanfly inhibit the proliferation of GMCs and showed no effect on apoptosis.The high dose of rapamycin (8-16 μg/L)could significantly increase the apoptotic rate of GMCs.Rapamycin could increase the mRNA and protein expression of p27 and p53. Conclusion Rapamycin can inhibit GMCs proliferation and promote GMCs apoptosis by increasing the expression of p27 and p53.
2.Correlation study between changes in intestinal microflora structure and immune indexes in newly treated patients with pulmonary tuberculosis
Jinhui XIE ; Rong YU ; Guomin SHI ; Xiaohua MA ; Sifang XIAO ; Yihang YI ; Ting ZHOU ; Yangen XIANG
Chinese Journal of Preventive Medicine 2021;55(12):1486-1490
To explore the correlation between the changes of the intestinal flora of newly treated pulmonary tuberculosis patients and the immune indicators of the body, and to provide a reference for the prevention and treatment of pulmonary tuberculosis. A single-center and case-control study was adopted. From October 2020 to April 2021, 43 patients with newly diagnosed tuberculosis in the Department of Tuberculosis, Affiliated Changsha Central Hospital,University of South China were selected as the control group. 43 cases of newly treated pulmonary tuberculosis (PTB), 43 healthy control (HC) during the same period, collected fresh feces and whole blood of subjects, and used Illumina Hiseq high-throughput sequencing technology to analyze 16S of all microorganisms in feces The V4 region of rRNA was amplified and sequenced, and the structure of the intestinal flora was analyzed by QIIME software. Use flow cytometry to determine the subject′s immune indicators (CD3 +, CD4 +, CD8 +, CD4 +CD25 +CD127 -Treg, CD14 +CD16 +, CD14 +CD16 -), and analyze the changes in intestinal flora and immune function in newly treated pulmonary tuberculosis patients Inherent connection. The χ2 test, t test, and Wilcox rank sum test were used to analyze the differences in age, gender, α diversity, and relative abundance of the two groups of people. Compared with the HC group, the alpha diversity of the intestinal flora in the PTB group decreased (shannon index: t=3.906, P=0.000 2; simpson index: Z=553, P=0.004 7; chao1 index: t=5.395, P=0.000 0). β diversity analysis showed that there were significant differences in the structure of the intestinal flora between the two groups ( P=0.000). Species difference analysis showed that at the phylum level, the relative abundance of Firmicutes in the PTB group was significantly lower than that in the HC group ( Z=486.0, P=0.000 5). At the genus level, there are 15 different bacterial genera between the two groups. In the PTB group, bifidobacterium, enterococcus, lactobacillus, anaerostipes, the relative abundance of the above 5 genera of veillonella is higher than that of the HC group ( P<0.05); Butyricimonas, clostridium, and broutella (blautia), coprococcus, dorea, lachnospira, roseburia, faecalibacterium, ruminococcus, the relative abundance of 10 bacterial genera including dialister was lower than that of the HC group ( P<0.05). Comparison of immune indexes between groups showed that CD14 +CD16 +monocytes (%) in the PTB group were higher than those in the HC group ( t=2.456, P=0.001 6<0.05), while CD14 +CD16 -monocytes (%) were lower than HC ( t=-4.368, P=0.000<0.05), while the differences in CD3 +, CD4 +, CD8 +, CD4 +/CD8 +and Treg (CD4 +CD25 +CD127 -) were not statistically significant ( P>0.05). Spearman correlation analysis showed that Firmicutes in the PTB group was negatively correlated with CD4 +/CD8 +, CD14 +CD16 +( r=-0.218, P=0.048; r=-0.245, P=0.025), and positively correlated with CD14 +CD16 -Correlation ( r=0.250, P=0.022); At the genus level, Faecalis is positively correlated with CD4 +/CD8 +and CD4 +( r=0.250, P=0.023; r=0.258, P=0.019); Rosella and CD3 +, CD8 +and CD14 +CD16 -are positively correlated ( r=0.27, P=0.024; r=0.219, P=0.046; r=0.027, P=0.039), and negatively correlated with CD14 +CD16 +( r=-0.280, P= 0.01). Changes in the structure of the intestinal flora of newly treated pulmonary tuberculosis patients may be one of the influencing factors of the immune function of the body. Targeted optimization of the structure of the intestinal flora and improvement of the body′s immunity may be used as an effective auxiliary treatment for pulmonary tuberculosis.
3.Correlation study between changes in intestinal microflora structure and immune indexes in newly treated patients with pulmonary tuberculosis
Jinhui XIE ; Rong YU ; Guomin SHI ; Xiaohua MA ; Sifang XIAO ; Yihang YI ; Ting ZHOU ; Yangen XIANG
Chinese Journal of Preventive Medicine 2021;55(12):1486-1490
To explore the correlation between the changes of the intestinal flora of newly treated pulmonary tuberculosis patients and the immune indicators of the body, and to provide a reference for the prevention and treatment of pulmonary tuberculosis. A single-center and case-control study was adopted. From October 2020 to April 2021, 43 patients with newly diagnosed tuberculosis in the Department of Tuberculosis, Affiliated Changsha Central Hospital,University of South China were selected as the control group. 43 cases of newly treated pulmonary tuberculosis (PTB), 43 healthy control (HC) during the same period, collected fresh feces and whole blood of subjects, and used Illumina Hiseq high-throughput sequencing technology to analyze 16S of all microorganisms in feces The V4 region of rRNA was amplified and sequenced, and the structure of the intestinal flora was analyzed by QIIME software. Use flow cytometry to determine the subject′s immune indicators (CD3 +, CD4 +, CD8 +, CD4 +CD25 +CD127 -Treg, CD14 +CD16 +, CD14 +CD16 -), and analyze the changes in intestinal flora and immune function in newly treated pulmonary tuberculosis patients Inherent connection. The χ2 test, t test, and Wilcox rank sum test were used to analyze the differences in age, gender, α diversity, and relative abundance of the two groups of people. Compared with the HC group, the alpha diversity of the intestinal flora in the PTB group decreased (shannon index: t=3.906, P=0.000 2; simpson index: Z=553, P=0.004 7; chao1 index: t=5.395, P=0.000 0). β diversity analysis showed that there were significant differences in the structure of the intestinal flora between the two groups ( P=0.000). Species difference analysis showed that at the phylum level, the relative abundance of Firmicutes in the PTB group was significantly lower than that in the HC group ( Z=486.0, P=0.000 5). At the genus level, there are 15 different bacterial genera between the two groups. In the PTB group, bifidobacterium, enterococcus, lactobacillus, anaerostipes, the relative abundance of the above 5 genera of veillonella is higher than that of the HC group ( P<0.05); Butyricimonas, clostridium, and broutella (blautia), coprococcus, dorea, lachnospira, roseburia, faecalibacterium, ruminococcus, the relative abundance of 10 bacterial genera including dialister was lower than that of the HC group ( P<0.05). Comparison of immune indexes between groups showed that CD14 +CD16 +monocytes (%) in the PTB group were higher than those in the HC group ( t=2.456, P=0.001 6<0.05), while CD14 +CD16 -monocytes (%) were lower than HC ( t=-4.368, P=0.000<0.05), while the differences in CD3 +, CD4 +, CD8 +, CD4 +/CD8 +and Treg (CD4 +CD25 +CD127 -) were not statistically significant ( P>0.05). Spearman correlation analysis showed that Firmicutes in the PTB group was negatively correlated with CD4 +/CD8 +, CD14 +CD16 +( r=-0.218, P=0.048; r=-0.245, P=0.025), and positively correlated with CD14 +CD16 -Correlation ( r=0.250, P=0.022); At the genus level, Faecalis is positively correlated with CD4 +/CD8 +and CD4 +( r=0.250, P=0.023; r=0.258, P=0.019); Rosella and CD3 +, CD8 +and CD14 +CD16 -are positively correlated ( r=0.27, P=0.024; r=0.219, P=0.046; r=0.027, P=0.039), and negatively correlated with CD14 +CD16 +( r=-0.280, P= 0.01). Changes in the structure of the intestinal flora of newly treated pulmonary tuberculosis patients may be one of the influencing factors of the immune function of the body. Targeted optimization of the structure of the intestinal flora and improvement of the body′s immunity may be used as an effective auxiliary treatment for pulmonary tuberculosis.
4.Study on the Difference of Volatile Components and Surface Color of Olibanum and Its Products
HUANG Ju ; XIE Sifang ; TANG Peili ; YANG Sicheng ; CHEN Shuhe ; YAN Jinsong ; SUN Wanjin ; DUAN Xueyun
Chinese Journal of Modern Applied Pharmacy 2023;40(18):2526-2534
OBJECTIVE To carry out qualitative and quantitative analysis of volatile components of Olibanum, Olibanum prepared with vinegar and Olibanum stir-fried with Rush by GC-MS, and the quantitative study of the surface color by RGB model, provide auxiliary reference for the subjective evaluation indexes of Olibanum and its products. METHODS The volatile oil of Olibanum, Olibanum prepared with vinegar and Olibanum stir-fried with Rush were extracted. Explored the difference of volatile components by GC-MS in combination with principal component analysis and orthogonal partial least-squares discriminant analysis make a comprehensive analysis. Collected the image information of Olibanum, Olibanum prepared with vinegar and Olibanum stir-fried with Rush, then measured the surface color of them by RGB color model, and counted three color differences. RESULTS The differences of odor among oils might be related to the contents and types of terpenoids and alcohols, especially linalool and 1-octanol. According to surface color determination results, Olibanum stir-fried with Rush was R*76.86%-85.49%, G*61.96%-70.59%, B*38.04%-45.88%; Olibanum prepared with vinegar was R*56.86%-61.57%, G*38.04%-41.96%, B*27.45%-30.59%; Olibanum was R*69.41%-74.51%, G*56.86%-62.35%, B*40.78%-47.06%. By t test, there were significant differences among the three color measurement results, which were consistent with the differences recorded in the corresponding standards for "yellow white" Olibanum, "yellow brown" Olibanum prepared with vinegar, and "golden yellow" Olibanum stir-fried with Rush, indicating that the model was accurate and reliable. CONCLUSION The differences of volatile components and surface color of Olibanum, Olibanum prepared with vinegar and Olibanum stir-fried with Rush are studied to provide reference for character description of Olibanum and its processed products, and to avoid errors caused by traditional subjective evaluation.