1.Identification and Molecular Analysis of H5N2 Subtype Highly Pathogenic Avian Influenza Virus Isolated From Goose
Chunguo LIU ; Ming LIU ; Yun ZHANG ; Dafei LIU ; Weiqi PAN ; Encheng SUN ; Jinling DU ; Hongtao LI
Progress in Biochemistry and Biophysics 2006;0(01):-
A H5N2 subtype avian influenza virus isolated from goose belongs to highly pathogenic avian influenza virus, and the intravenous pathogenicity indexes (IVPI) =2.99. But ducks are not sensitive to this isolated influenza virus. The virus can infect mouse but only replicates in lung and has no pathogenicity. HA and NA gene of this isolated strain share 99.4% and 99.8% nucleotide sequence identity to the HA gene of A/chicken/Hubei/ 489/2004 (H5N1) and the NA gene of A/chicken/Jilin/53/01 (H9N2), and share 99.3% and 99.6% amino acid sequence identity to the HA protein of A/chicken/Hubei/489/2004 (H5N1), A/swan/Guangxi/307/2004 (H5N1), A/wild duck/ Guangdong/314/2004(H5N1), A/chicken/Henan/210/2004(H5N1) and the NA protein of A/chicken/ Jilin/53/01 (H9N2). There are several continuous basic amino acids (-RRRKKR-) at the cleavage site of HA protein. Phylogenetic trees analysis of HA and NA gene suggests that the isolated influenza virus probably originated from the reassortment of H5N1 and H9N2 subtype influenza virus.
2.Study of Alzheimer′s Disease Biomarkers Based onSerum Peptidomics
Xiangyi KONG ; Jianshi DU ; Ming MA ; Jinling XU ; Shuiming LI ; Yong WANG ; Qing ZHAO
Chinese Journal of Analytical Chemistry 2017;45(7):937-943
Early diagnosis and intervention is an important way to delay the progress of Alzheimer′s disease (AD).Compared with cerebrospinal fluid, blood sampling is not invasive and easy to be obtained in clinic practice.In this study, the serum samples of 9 controls, 10 AD and 12 mild cognitive dysfunction (MCI) patients were analyzed and compared through one by one analysis to screen potential markers for AD diagnosis.The experimental results showed that VGFYESDVMGR of α-2-macroglobulin peptide was closely related to the late stage of AD disease, and the large amount degradation of apolipoprotein C-Ⅲ, histone H1.2 and histone H1.4 was significantly related to early stages of AD progression.The characteristics of serum peptidome were different for the early and late AD, and these four proteins may be used as potential biomarkers of AD disease.In addition, the obvious ladder sequence characteristic was observed for apolipoprotein C-Ⅲ and histone H1, which could partly explain why the peptides distribution in different samples was somewhat contingent.On the contrary, the distribution at protein level was more stable.Finally, it was confirmed that the peptides of proteins such as fibrinogen α-chain, thymosin β-4 and patchy proteins were the dominant peptides in all serum samples.Overall, this study showed that the method of using serum peptidomics to diagnose AD was possible.The results may provide evidence and references for the large-scale clinical validation of AD.
3.Downregulation of lanosterol synthase gene expression by antisense RNA technology in Saccharomyces cerevisiae.
Qinghua WANG ; Lili GAO ; Huichao LIANG ; Guohua DU ; Ting GONG ; Jinling YANG ; Ping ZHU
Acta Pharmaceutica Sinica 2015;50(1):118-22
The cyclization of 2,3-oxidosqualene is the key branch point of ergosterol and triterpenoid biosynthesis. Downregulation of 2,3-oxidosqualene metabolic flux to ergosterol in Saccharomyces cerevisiae may redirect the metabolic flux toward the triterpenoid synthetic pathway. In our study, primers were designed according to erg7 gene sequence of S. cerevisiae. Three fragments including 5' long fragment, 5' short fragment and erg7 coding region fragment were amplified by PCR. 5' long fragment consists of the promoter and a part of erg7 coding region sequence. 5' short fragment consists of a part of promoter and a part of erg7 coding region sequence. These fragments were inserted reversely into pESC-URA to construct antisense expression plasmids. The recombinant plasmids were transformed into S. cerevisiae INVSc1 and recombinant strains were screened on the nutritional deficient medium SD-URA. The erg7 expression level of recombinant strains, which harbored antisense expression plasmid of erg7 coding region, was similar to that of INVScl by semi-quantitative PCR detection. But erg7 expression level of recombinant strains, which harbored 5' long antisense fragment and 5' short antisense fragment, was significantly lower than that of the control. The results of TLC and HPLC showed that the ergosterol content of recombinant strains, which harbored 5' long antisense fragment, decreased obviously. The ergosterol contents of the others were almost equal to that of INVSc1. Lanosterol synthase gene expression was downregulated by antisense RNA technology in S. cerevisiae, which lays a foundation for reconstructing triterpenoid metabolic pathway in S. cerevisiae by synthetic biology technology.
4.The basic strategies and research advances in the studies on glycosyltransferases involved in ginsenoside biosynthesis.
Huichao LIANG ; Qinghua WANG ; Ting GONG ; Guohua DU ; Jinling YANG ; Ping ZHU
Acta Pharmaceutica Sinica 2015;50(2):148-53
Traditional herbal medicines, Panax ginseng, Panax quinquefolium and Panax notoginseng, attract our attention for their extensive and powerful pharmacological activities. Ginsenosides are the main active constituents of these medicinal herbs. The related glycosyltransferases involved in ginsenoside biosynthesis are the key enzymes which catalyze the last important step. Modification of ginsenoside aglycones by glycosyltransferases produces the complexity and diversity of ginsenosides, which have more extensive pharmacological activity. At present, ginsenoside aglycones and compound K have been obtained by synthetic biology. As the last step of ginsenoside biosynthesis, glycosylation of ginsenoside aglycones has been studied intensively in recent years. This review summarizes the basic strategies and research advances in studies on glycosyltransferases involved in ginsenoside biosynthesis, which is expected to lay the theoretical foundation for the in-depth research of biosynthetic pathway of ginsenosides and their production by synthetic biology.
5.Effects of 2 Gy γ-irradiation on the number of regulatory T cells and Thl7 cells and immune balance in mice
Xiaoyun YANG ; Li DU ; Qiong MA ; Xiujie PAN ; Jinling CAI ; Fengjun XIAO ; Yufang CUI
Chinese Journal of Radiological Medicine and Protection 2013;(1):6-9
Objective To observe the effects of 2 Gy γ-ray irradiation on regulatory T cells (Tregs) and Th17 cells and immune balance of mice.Methods A total of fifty C57BL/6 male mice were randomly divided into two groups,the irradiated group exposed to 2 Gy of whole body γ-ray irradiation,and the control group sham-irradiated.At 1,3,7,14 and 28 d after radiation,changes of peripheral haemogram were detected respectively and Tregs in peripheral blood,thymus and spleen and Th17 cells in spleen were analyzed by flow cytometry.Results Compared with control group,the number of peripheral blood white cells (WBC) and lymphocyte in irradiated group reduced significantly post-irradiation (t =8.89-33.54,P < 0.05),while the cell number of peripheral CD4 + CD25 + Tregs post-irradiation rose but not significantly.Thymic Treg cells increased 1 and 3 d post-irradiation(t =-6.45,-10.59,P <0.05),but reduced 28 d post-irradiation (t =5.34,P < 0.05).Splenic Treg cells ascended obviously from 1 to 14 d post-irradiation (t =-6.82-3.89,P < 0.05).After irradiation splenic Th17 cells increased at 1 d,and reached the maximal level at 3 d (t =-2.42,P < 0.05),more obviously than splenic Treg cells.The reduction of Treg/Th17 ratio from 1 to 14 d post-irradiation disturbed Treg/Th17 balance and made it drift to the direction of Th17 (t =4.02-8.04,P < 0.05).Conclusions Treg/Th17 imbalance plays an important role in immune injury induced by irradiation.
6.Correlation analysis on total lymphocyte count and CD4 count in HIV-infected patients: A retrospective evaluation.
Yuming, WANG ; Shuying LIANG ; Erman, YU ; Jinling, GUO ; Zizhao, LI ; Zhe, WANG ; Yukai, DU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(5):712-6
CD4 count is the standard method for determining eligibility for highly active antiretroviral therapy (HAART) and monitoring HIV/AIDS disease progression, but it is not widely available in resource-limited settings. This study examined the correlation between total lymphocyte count (TLC) and CD4 count of HIV-infected patients before and after HAART, and assessed the thresholds of TLC for making decisions about the initiation and for monitoring HAART. A retrospective study was performed, and 665 HIV-infected patients with TLC and CD4 count from four counties (Shangcai, Queshan, Shenqiu and Weishi) were included in the study. Pearson correlation and receiver operating characteristic (ROC) were used. TLC and CD4 count after HAART was significantly increased as compared with pre-HAART (P<0.01). An overall positive correlation was noted between TLC and CD4 count (pre-HAART, r=0.73, P=0.0001; follow-up HAART, r=0.56, P=0.0001). The ROC curve between TLC and CD4 count showed that TLC ≤ 1200 cells/mm(3) could predict CD4 < 200 cells/mm(3) with a sensitivity of 71.12%, specificity of 66.35% at pre-HAART. After 12-month HAART, the optimum prediction for CD4 count < 200 cells/mm3 was a TLC ≤ 1300 cells/mm(3), with a sensitivity of 63.27%, and a specificity of 74.84%. Further finding indicated that TLC change was positively correlated to CD4 change (r=0.77, P=0.0001) at the time point of 12-month treatment, and the best prediction point of TLC change for CD4 increasing was 135 cells/mm(3). TLC and its change can be used as a surrogate marker for CD4 count and its change of HIV-infected individuals for making decisions about the initiation and for monitoring HAART in resource-limited settings.
7. The mediating role of mental capital in understanding social support and job satisfaction
Zhongchen LI ; Kang WANG ; Xinyi ZHU ; Jinling DU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2018;36(8):594-597
Objective:
To understand the relationship between perceived social support, psychological capital and job satisfaction, and analyze the mediating effect of psychological capital between perceived social support and job satisfaction.
Methods:
In 2017, June to July, we use cluster random sampling method to evaluate 888 medical staff from 5 hospitals in Jinan and Yantai in Shandong provience by Perceived Social Support Scale、PsyCap Questionnaire and MinnesotaSatisfaction Questionnaire.
Results:
Correlation analysis showed that perceived social support was significantly positively correlated with self-efficacy, hope, resilience, optimism and job satisfaction (
8.Effect of Schisandra chinensis on interleukins, glucose metabolism, and pituitary-adrenal and gonadal axis in rats under strenuous swimming exercise.
Jie LI ; Jian WANG ; Jia-Qing SHAO ; Hong DU ; Yang-Tian WANG ; Li PENG
Chinese journal of integrative medicine 2015;21(1):43-48
OBJECTIVETo investigate the effect of Chinese medicine (CM) Schisandra chinensis on interleukin (IL), glucose metabolism, and pituitary-adrenal and gonadal axis of rats after strenuous navigation and exercise.
METHODSA total of 45 Sprague-Dawley rats were randomized into the quiet control group, the stress group, and the CM group (15 in each group). The CM group received 2.5 g/kg of Schisandra chinensis twice per day for one week before modeling. Except the quiet controls, rats were trained using the Bedford mode for 10 days. On the 11th day, they performed 3 h of stressful experimental navigation and 3 h of strenuous treadmill exercise. The levels of serum testosterone (T), cortisol (CORT), luteinizing hormone (LH), IL-1, IL-2, and IL-6 were tested by radioimmunoassay and enzyme-linked immunosorbent assay, respectively. The adrenal cortex ultrastructure was observed using electron microscopy.
RESULTSCompared with the quiet control group, after navigation and strenuous exercise, blood glucose was increased, and T level was decreased in the stress group (both P<0.01). The blood glucose, CORT, IL-1 and IL-2 levels were significantly reduced in the CM group (P<0.05 or P<0.01) as compared with the stress group. Electron microscopy revealed that the rats in the CM group had a smaller decrease in adrenal intracellular lipid droplets and higher levels of apoptosis than those in the stress group.
CONCLUSIONSSchisandra chinensis can reduce serum CORT and blood glucose levels in stressed rats. It appears to protect the cell structure of the adrenal cortex, and offset the negative effects of psychological stress and strenuous exercise related to immune dysfunction. Schisandra chinensis plays a regulatory role in immune function, and can decrease the influence of stress in rats.
Adrenal Cortex ; pathology ; ultrastructure ; Animals ; Blood Glucose ; metabolism ; Glucose ; metabolism ; Gonads ; drug effects ; metabolism ; Hydrocortisone ; blood ; Interleukin-1 ; blood ; Interleukin-2 ; blood ; Interleukin-6 ; blood ; Interleukins ; blood ; Luteinizing Hormone ; blood ; Male ; Physical Conditioning, Animal ; Pituitary-Adrenal System ; drug effects ; metabolism ; Plant Extracts ; pharmacology ; Rats, Sprague-Dawley ; Schisandra ; chemistry ; Swimming ; physiology ; Testosterone ; blood
9.Correlation Analysis on Total Lymphocyte Count and CD4 Count in HIV-infected Patients: A Retrospective Evaluation
WANG YUMING ; LIANG SHUYING ; YU ERMAN ; GUO JINLING ; LI ZIZHAO ; WANG ZHE ; DU YUKAI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(5):712-716
CD4 count is the standard method for determining eligibility for highly active antiretroviral therapy (HAART) and monitoring HIV/AIDS disease progression,but it is not widely available in resource-limited settings.This study examined the correlation between total lymphocyte count (TLC) and CD4 count of HIV-infected patients before and after HAART,and assessed the thresholds of TLC for making decisions about the initiation and for monitoring HAART.A retrospective study was performed,and 665 HIV-infected patients with TLC and CD4 count from four counties (Shangcai,Queshan,Shenqiu and Weishi) were included in the study.Pearson correlation and receiver operating characteristic (ROC) were used.TLC and CD4 count after HAART was significantly increased as compared with pre-HAART (P<0.01).An overall positive correlation was noted between TLC and CD4 count (pre-HAART,r=0.73,P=0.0001; follow-up HAART,r=0.56,P=0.0001).The ROC curve between TLC and CD4 count showed that TLC ≤ 1200 cells/mm3 could predict CD4 < 200 cells/mm3 with a sensitivity of 71.12%,specificity of 66.35%at pre-HAART.After 12-month HAART,the optimum prediction for CD4 count < 200 cells/mm3 was a TLC ≤ 1300 cells/mm3,with a sensitivity of 63.27%,and a specificity of 74.84%.Further finding indicated that TLC change was positively correlated to CD4 change (r=0.77,P=0.0001) at the time point of 12- month treatment,and the best prediction point of TLC change for CD4 increasing was 135 cells/mm3.TLC and its change can be used as a surrogate marker for CD4 count and its change of HIV-infected individuals for making decisions about the initiation and for monitoring HAART in resource-limited settings.
10. Correlation of the thyroid hormone level with glycemic fluctuation in patients with euthyroid type 2 diabetes mellitus
Fei WU ; Jun-xiu CHEN ; Zhou-qin ZHENG ; Rui-feng WANG ; Hong DU
Journal of Medical Postgraduates 2018;31(12):1272-1275
Objective Diabetes mellitus (DM) is often complicated by thyroid hormone abnormality. The aim of this study was to investigate the relationship between the thyroid hormone level and glycemic fluctuation in patients with euthyroid type 2 DM (T2DM).Methods A total of 143 euthyroid T2DM patients were treated in the Department of Endocrinology of Nanjing General Hospital from January 2014 to December 2015. The continuous blood glucose monitoring system was used for 72-hour continuous monitoring of blood glucose fluctuation indexes, including the standard deviation (SD) of the glucose level, the mean amplitude of glycemic excursions (MAGE), and the absolute mean of daily differences (MODD). According to the percentile of the MAGE level, the patients were divided into groups Q1 (MAGE<4.1864, n=35), Q2 (4.1864≤MAGE<5.3764, n=37), Q3 (5.3764≤MAGE<6.8484, n=35), and Q4 (MAGE≥6.8484, n=36), compared the thyroid hormone level and such thyroid function parameters as serum free triiodothyronine (FT3), triiodothyronine (T3), free thyroxine (FT4) and FT3/FT4 among the four groups, and analyzed the correlation between the thyroid hormone level and glycemic indexes.Results The levels of serum FT3, T3 and FT3/FT4 were decreased with the increase of MAGE (P<0.05), and those of FT3 and FT3/FT4 were 4.11±0.77 and 0.38±0.37 in group Q1, 4.06±0.55 and 0.34±0.37 in Q2, 3.49±0.57 and 0.33±0.06 in Q3, and 3.68±0.65 and 0.31±0.09 in Q4, with statistically significant differences between any two groups (P<0.05). There were also statistically significant differences among the four groups (P<0.05) in the T3 and FT4 levels (P<0.05). The levels of FT3 and T3 were correlated negatively with SD, MODD and MAGE (P<0.05), and so was that of FT3/FT4 with MAGE (r=-0.243, P<0.05).Conclusion MAGE reduces the levels of FT3 and FT3/FT4 in patients with euthyroid T2DM probably by inhibiting the conversion of T4 to T3.