2.Construction of Recombinant Adenovirus Expression Vector of Human Sema4C Gene and Its Expression in Mouse Myoblasts Cell Line C2C12
Hai-Tao WU ; Shu-Hong LIU ; Yan WU ; Jun-Die FAN ; Wen-Hong FAN ; Ming FAN ;
China Biotechnology 2006;0(04):-
To generate recombinant adenovirus expression vector of human Sema4C gene and observe its expression in mouse myoblasts cell line C2C12 for ensuring easy access to investigate the role of Sema4C gene during myogenesis. The recombinant plasmid was packaged and amplified after being transfected in HEK293 cells through Lipofectamine. After infecting C2C12 myoblasts with recombinant adenovirus vector, the adenoviral infection efficiency was determined by confocal microscope which showed that the expression of green fluorescence could be detected at 12h and then reached peak at 24h after recombinant adenovirus infection. The infection efficiency was almost 100% confirmed by FACS examination. Detection of WB indicated that the expression of Sema4C in C2C12 of recombinant adenoviral infection group was significantly higher than that of the control group (P
3.Oligonucleotide array of genomic expression in complete androgenic hydafidiform mole
Jianyun XU ; Feng YE ; Wei-Guo LV ; Die HONG ; Jian-Hua QIAN ; Xing XIE ;
Chinese Journal of Obstetrics and Gynecology 2001;0(08):-
Objective To compare genomic expression differences between androgenic complete hydatidiform mole (AnCHM) and normal first trimester villi with similar gestation weeks,and search for potential adjuvant diagnostic molecular markers.Methods Short tandem repeat (STR) detection was used to identify AnCHM,human oligonucleotide array U133 Plus 2.0 was used to measure genomic expression differences between AnCHM and normal villi,and quantitative fluorescent RT-PCR was used to verify array of several genes.Results Nine of 11 histologically diagnosed complete hydatidiform moles were found to be AnCHM by means of STR,and the other 2 were biparental complete hydatidifonn mole (BiCHM). Compared with villi,oligonueleotide array showed 279 genes (0.72%,279/38 500) were over expressed and 1710 genes (4.44%,1710/38 500) under expressed in AnCHM.Bioinformatics analysis found that differentially expressed genes were involved in multiple biological processes and pathways.Changes of imprinting genes,growth hormone genes and chorionie somatomammotropin hormone genes were especially remarkable.Conclusions Pathogenesis of AnCHM is a complex process involving multiple genes and pathways.Altered expression of imprint genes may play important roles in the process.
4.Combined effects of triclosan and PCB153 on the activity of superoxide dismutase and the content of malondialdehyde in zebrafish liver
Cheng ZHOU ; Nanxiang WU ; Hongliang FAN ; Ye YANG ; Ming GAO ; Die CHEN ; Hong SHEN
Journal of Preventive Medicine 2019;31(4):330-334
Objective :
To investigate the combined effects of triclosan(TCS)and PCB153 on the activity of superoxide dismutase(SOD)and the concentration of malondialdehyde(MDA)in zebrafish liver.
Methods :
Adult zebrafish were exposed to a series of concentrations of TCS,and the mortality in each group was observed and recorded during the acute toxicity test process. The concentrations in subsequent combined exposure experiments were arranged on the basis of the 96 h-LC50. The factorial design was used to determine the concentrations of combined exposure groups between TCS(0,0.125,0.5 μmol/L)and PCB153(0,0.05,0.2 μmol/L). After 5,10 and 14 days of exposure,the zebrafish livers were dissected and frozen in each group. The potential interactions of these two compounds were analyzed according to the results of the SOD and MDA.
Results :
The 96 h-LC50 of TCS exposed to adult zebrafish was 2.64 μmol/L(95%CI:2.37-2.89 μmol/L). After 5 days of exposure,combined exposure to 0.5 μmol/L TCS+0.2 μmol/L PCB153 caused lower liver SOD activities than single exposure groups and the control group(P<0.05). After 10 days of exposure,combined exposure to 0.125 μmol/L TCS+0.05 μmol/L PCB153,0.5 μmol/L TCS+0.05 μmol/L PCB153 caused lower liver SOD activities than single exposure groups and the control group(P<0.05). After 14 days of exposure,combined exposure to 0.5 μmol/L TCS+0.05 μmol/L PCB153,0.5 μmol/L TCS+0.2 μmol/L PCB153 caused higher liver SOD activities than single exposure groups and the control group(P<0.05). There was an interactive effect between TCS and PCB153 on the liver SOD activity in zebrafish(P<0.05). There was no significant effect of MDA content in each group.
Conclusion
Combined exposure to TCS and PCB153 could enhance (inhibit first) the liver SOD activities in zebrafish,and the interaction was synergistic.
5.Comparative study on biochemical characteristic of Yersinia pestis from some natural focuses of plague in Yunnan Province
Ying, GUO ; Die-xin, WEI ; Yun, LIANG ; Peng, SU ; Zhi-ming, YANG ; Shan-shan, DONG ; You-hong, ZHONG ; Zhi-zhong, SONG
Chinese Journal of Endemiology 2009;28(4):373-375
Objective To compare the difference of biochemical characteristics and virulent Pst Ⅰ of Yersinia pestis strains in traditional focuses of plague in Yunna Province and in the new focuses of plague in Yulong County. Methods The identification data of biochemical characteristics(Rhamnose, Glycerol, Maltose, L-Arabina and Melibiose fermentation) and virulence factor(Pst Ⅰ) from different focuses of plague in Yunna Province were Retrospectively collected by tube test followed by the analysis using statistics software SAS 8.0 by Fisher exact probability of disordered two-way R × C table χ2 test. Results Among 48 strains of Yersinia pestis from hantaan type plague focus, 1 strain fermented L-maltose, 48 strains fermented Glycerol. Among 165 strains of Yersinia pestis from the Soul type plague focus, 1 strain did not ferment L-maltose, only one of them fermented Glycerol. 1 strain from the Soul type plague focus was confirmed to have mutation, for the test of nitrate reduction reaction was negative. All 5 strains of Yersinia pestis from the new focuses of plague in Yulong County fermented L-maltose and Glycerol. The statistical result showed that the differences in L-maltose and Glycerol fermentation of Yersinia pestis from different natural focuses of plague in Yunnan Province were statistically siguificant (P < 0.01). The differences of other biochemical characteristics and Pst Ⅰ were not statistically significant (P > 0.01). Conclusions Biochemical characteristics of Yersinia pestis from the hantaan type plague focus and the Soul type plague focus in Yunnan province are overlapping. Biochemical characteristics of Yersinia pestis from the new focuses of plague in Yulong County are different from those tradition focuses of plague in Yunna Province but share similarities to those from Unquiculatus focuses in North Tibet.
6. Research progress in boswellic acids from medicinal plants in Boswellia Roxb. ex Colebr. and their pharmacological action as well as mechanisms
Chinese Traditional and Herbal Drugs 2016;47(12):2175-2181
Frankincense, the gum resin from the plants in Boswellia Roxb. ex Colebr. or the other plants belonging to Burseraceae, has a long history as a medicine in China. The investigations on the pharmacological effects of the resin have demonstrated its anti-inflammatory, antisepsis, antitumor effects, etc. This article classifies and summarizes the preparation methods, pharmacological effects, action mechanisms, and metabolic process in vivo or in vitro of boswellic acids compounds, and providing the reference for the further development and utilization of boswellic acids and medicinal plants in Boswellia Roxb. ex Colebr.
7.Application of the theories of materiomic release kinetics to the evaluation of the sustained release kinetics and synchronicity of yinqiaojiedu honeyed pills.
Die LING ; Ji-Wen ZHANG ; Li-Bing CHEN ; Meng LIN ; Wei-Hong GE ; Jing-Kai GU
Acta Pharmaceutica Sinica 2008;43(11):1140-1146
Yinqiaojiedu honeyed pills were equally divided into 1/4, 1/8, 1/12, and 1/16 parts. The materiomics release rates within 12 h of the intact Yinqiaojiedu honeyed pills and the divided granules were determined by the paddle method with a rotate speed at 100 r x min(-1), and the materiome was quantified by UV-scan and Kalman filter methods. The intact Yinqiaojiedu honeyed pills behaved typical sustained release profiles, while the well-divided portions also maintained a sustained release profile over 2-4 h. The release rates were well correlated with the extents for the divisions of the pills. The Weibull distribution parameters, Td and T50, were reduced in line with the particle size, indicating that the ways of administration of the pills may play a role in the in vivo pharmacokinetics of the pills. The visualization results showed obvious difference of materiomic release synchronicities between the intact pills and the equally divided particles, and the divisions enhanced the asynchronization. Therefore the novel theory of materiomic release/dissolution kinetics of traditional Chinese medicines (TCMs) quantitatively proved the traditional dosage form, namely, honeyed pills, as a prototype of the sustained-release dosage form with a visualization of the scientific connotation to the old saying in the classics of TCM, Pills, the moderate ones in action. In terms of materiome increase for each period of the release profiles, the materiomic release synchronicity was visually demonstrated. The novel theories provided methodological basis for the evaluation of traditional dosage forms and the design of the modern drug delivery systems for TCMs.
Delayed-Action Preparations
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Drug Combinations
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Drug Delivery Systems
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Drugs, Chinese Herbal
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administration & dosage
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chemistry
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isolation & purification
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pharmacokinetics
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Kinetics
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Medicine, Chinese Traditional
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Particle Size
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Plants, Medicinal
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chemistry
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Solubility
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Technology, Pharmaceutical
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methods
8.Hydrogen sulfide induces apoptosis of pulmonary artery smooth muscle cell in rats with pulmonary hypertension induced by high pulmonary blood flow.
Wei LI ; Hong-Fang JIN ; Die LIU ; Jing-Hui SUN ; Pei-Jun JIAN ; Xiao-Hui LI ; Chao-Shu TANG ; Jun-Bao DU
Chinese Medical Journal 2009;122(24):3032-3038
BACKGROUNDAbnormal apoptosis of pulmonary artery smooth muscle cells (PASMCs) is an important pathophysiological process in the pulmonary artery structural remodeling and pulmonary hypertension. We investigated possible effect of endogenous hydrogen sulfide (H2S) on apoptosis of PASMCs during the development of pulmonary hypertension induced by high pulmonary blood flow.
METHODSThirty-nine male Sprague-Dawley rats were randomly assigned to 4-week control, 4-week shunt, 4-week shunt + propargylglycine (PPG), 11-week control, 11-week shunt and 11-week shunt + sodium hydrosulfide (NaHS) groups. Rats in 4-week shunt, 4-week shunt + PPG, 11-week shunt and 11-week shunt + NaHS groups underwent an abdominal aorta-inferior vena cava shunt. Rats in 4-week shunt + PPG group were intraperitoneally injected with PPG, an inhibitor of endogenous H2S production, for 4 weeks. Rats in 11-week shunt + NaHS group were intraperitoneally injected with NaHS, a H2S donor, for 11 weeks. Lung tissue H2S was evaluated by sulfide-sensitive electrode. Apoptosis of PASMCs were detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL). Expressions of Fas, bcl-2 and caspase-3 in the PASMCs were analyzed with immunochemical staining.
RESULTSFour weeks after the shunting operation, the apoptosis of PASMCs and expression of Fas and caspase-3 were significantly decreased (P < 0.01), but expression of bcl-2 increased significantly (P < 0.01). PPG administration further inhibited the apoptosis of PASMCs, downregulated the expression of Fas and caspase-3 (P < 0.01), but increased the expression of bcl-2 (P < 0.01). After 11 weeks of shunting operation, the apoptosis of PASMCs and expression of Fas and caspase-3 were significantly decreased (P < 0.01), but expression of bcl-2 increased obviously (P < 0.01). NaHS administration significantly increased the apoptosis of PASMCs, upregulated the expression of Fas and caspase-3, but inhibited the expression of bcl-2.
CONCLUSIONSH2S induces the apoptosis of PASMCs in the development of high pulmonary blood flow-induced pulmonary hypertension by activating the Fas pathway and inhibiting the bcl-2 pathway.
Alkynes ; pharmacology ; Animals ; Apoptosis ; drug effects ; Blood Flow Velocity ; physiology ; Blotting, Western ; Glycine ; analogs & derivatives ; pharmacology ; Hemodynamics ; drug effects ; Hydrogen Sulfide ; pharmacology ; Hypertension, Pulmonary ; etiology ; physiopathology ; Immunohistochemistry ; In Situ Nick-End Labeling ; Male ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Pulmonary Artery ; cytology ; Random Allocation ; Rats ; Rats, Sprague-Dawley
9.Association of Visit-to-Visit Variability of Blood Pressure with Cardiovascular Disease among Type 2 Diabetes Mellitus Patients: A Cohort Study
Zhe Bin YU ; Die LI ; Xue Yu CHEN ; Pei Wen ZHENG ; Hong Bo LIN ; Meng Ling TANG ; Ming Juan JIN ; Jian Bing WANG ; Kun CHEN
Diabetes & Metabolism Journal 2019;43(3):350-367
BACKGROUND: Increasing evidence has shown that visit-to-visit variability (VVV) of blood pressure (BP) is associated with an increased risk of cardiovascular disease (CVD). The objective of this study was to evaluate the impact of VVV of systolic blood pressure (SBP) and diastolic blood pressure (DBP) on the risk of CVD among patients with type 2 diabetes mellitus (T2DM) in China. METHODS: We conducted a retrospective cohort study of 10,163 T2DM patients who were not previously diagnosed with CVD from January 2008 to December 2012 in Ningbo, China. The VVV of BP was calculated using five metrics, including standard deviation (SD), coefficient of variation (CV), variation independent of mean, average real variability, and successive variability (SV) of measurements, obtained over a 24-month measurement period. Hazard ratios and 95% confidence intervals (CIs) were estimated by Cox proportional hazards regression models for the associations of variability in BP with risk of CVD. RESULTS: A total of 894 CVD events were observed during a median follow-up of 49.5 months. The hazard ratio in the highest quintile of SD of SBP was 1.24 (95% CI, 1.01 to 1.52) compared with patients in the lowest quintile. The association between higher VVV of DBP and risk of CVD was not consistent across different metrics and sensitivity analyses. CONCLUSION: Higher VVV of SBP was associated with an increased risk of CVD, irrespective of the mean SBP level. Future studies are needed to confirm these findings.
Blood Pressure
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Cardiovascular Diseases
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China
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Cohort Studies
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Diabetes Mellitus, Type 2
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Follow-Up Studies
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Humans
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Retrospective Studies
10.Effect of Salviae Miltiorrhizae Radix et Rhizoma on diversity of intestinal flora in diabetic nephropathy rats.
Hong-Die CAI ; Shu-Lan SU ; Jian-Ming GUO ; Jin-Ao DUAN
China Journal of Chinese Materia Medica 2021;46(2):426-435
This study aimed to explore the effect of Salviae Miltiorrhizae Radix et Rhizoma, its stems and leaves on the diversity of intestinal microflora in rats with diabetic kidney injury. Diabetic rats model was established by feeding high glucose and high fat diet and 5% glucose solution with intraperitoneal injection of 30 mg·kg~(-1) streptozocin(STZ). The rats were randomly divided into normal group, model group, irbesartan control group, Huangkui Capsules control group, as well as low, middle and high dose groups of Sal-viae Miltiorrhizae Radix et Rhizoma, its stems and leaves. After administration for 2 weeks, 16 S rRNA technique was used to analyze the diversity of intestinal microflora in the feces of each group. The results showed rats in the model group developed renal tubular epithelial vacuole degeneration and a large amount of inflammatory cell infiltration in the renal interstitium. A small amount of inflammatory cell infiltration was seen in each administration group. The kidney structure of rats in irbesartan group, Huangkui Capsules group, high-dose group of Salviae Miltiorrhizae Radix et Rhizoma and its stem water extract, as well as high dose group of Salviae Miltiorrhizae Radix et Rhizoma and its stem ethnol extract group was close to the normal group. The diversity and structure of intestinal flora in the model group were significantly different from those in the normal group. Each administration group improved the fecal flora diversity in rats with diabetic kidney injury to a certain extent, especially the high dose of Salviae Miltiorrhizae Radix et Rhizoma and its stems water extract. Different flora were found in feces of diabetic nephropathy model rats on class, order, family and genus levels. On families and genera levels, the relative abundance of Bifidobacterium, Turicibacter, Peptostreptococcaceae, Desulfovibrio, and SMB53 showed an upward trend in model group, but that of Lactobacillus, Clostridium, Rikenella, Rumen fungi showed a downward trend. The administration groups can improve the relative abundance of the above intestinal flora in the model rats to a normal-like level. The results of this study provide a reference for resource utilization and further development of Salviae Miltiorrhizae Radix et Rhizoma.
Animals
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Diabetes Mellitus, Experimental/drug therapy*
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Diabetic Nephropathies/drug therapy*
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Drugs, Chinese Herbal
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Gastrointestinal Microbiome
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Rats
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Salvia miltiorrhiza