1.Effect of dipsacus on the expression of β-amyloid in parietal and hippocampus of Alzheimer's disease rats
Wenbo TAN ; Depei YUAN ; Fuming YANG
Chinese Journal of Geriatrics 2013;(2):212-214
Objective To study the effect of dipsacus on the expression of β-amyloid in parietal cortex and hippocampus of Alzheimer's disease model rats.Methods A total of 40 rats were randomly divided into control group,model group,dipsacus group and vitamin E group (n=10,each)General situation of rats was observed after 42 days of dipsacus treatment.The ability of learning and memory in rats was tested by Morris water maze.The content of β-amyloid in parietal cortex and hippocampus of each group rats were measured using double antibody sandwich method.Results Model group compared with control group showed that general conditions deteriorated markedly;escaping delitescence was prolonged significantly(P<0.01) ; the activity time on the former platform quadrant reduced significantly (t=4.6261,P=0.0002) ; the number of crossing the former platform reduced significantly (t=6.5335,P=0.0000) ; the content of Aβ in parietal cortex and hippocampus increased significantly (t=4.2812,P=0.0004) (t=5.2499,P=0.0001).Dipsacus group and vitamin E group compared with model group showed that general condition improved significantly;escape latency was shortened significantly (P< 0.01); the activity time on the former platform quadrant increased significantly [(45.76±12.15) s,(48.70±10.25) s and (30.20±10.48) s (t=3.0666,3.9908,P=0.0066,0.0009)]; the number of crossing the former platform increased significantly [(3.02±1.19) t/2min,(3.56±0.85) t/2min and (1.43 ± 1.24) t/2min (t=2.9256,4.4804,P=0.0090,0.0003)]; the content of Aβ in parietal cortex and hippocampus reduced significantly in parietal cortex [(280.37-51.40) pg/g,(263.14 ± 45.52) pg/g versus (337.46 ±70.51) pg/g (t=2.1164,2.8003,P=0.0485,0.0118)],in hippocampus [(295.60±67.58)pg/g,(274.38±57.56)pg/g versus (388.26±83.72)pg/g(t=2.9256,4.4804,P=0.0090,0.0003)].Conclusions Dipsacus can reduce expression of Aβ in brain areas related to general intelligence that may have anti-Alzheimer's disease action.The active ingredients of dipsacus may be natural vitamin E.
2.Determination of vancomycin in human serum by in situ formed ionic liquids microextraction-ultra performance liquid chromatography (ISFILM-UPLC) methods
Dejin XU ; Wenbo YUAN ; Juan LYU
Journal of China Pharmaceutical University 2017;48(2):196-200
A method based on in situ formed ionic liquids microextraction-ultra performance liquid chromatography(ISFILM-UPLC) was established for the determination of vancomycin in human serum.The samples were pretreated by vortex and centrifugation.[C6MIM] [Br] and NaPF6 were applied as the extracting agent,and Metronidazole as the internal standard.Methanol was applied to redissolve the precipitate.The separation was carried out on a BEH C18 column,and the mobile phase consisted of methanol and 0.05 mol/L monopotassium phosphate solution.The flow rate was 0.2 mL/min,the injection volume was 5 μL,and the column temperature was 18 ℃.The key parameters of this method such as the specificity,accuracy and precision were validated.This method showed good correlation with the immunoassay.It shows great potential for the application in clinical practice.
3.The effect of SiRNA -Oct4 expression on HCC cell line HepG2
Wenbo ZHOU ; Fangjun YUAN ; Can ZOU ; Zhengpeng ZHU ; Jianbo MA ; Zongqing DAI ; Youshun ZHANG
Chinese Journal of General Surgery 2011;26(6):467-469
Objective To investigate the expression of Oct4 in liver cancer, and the interrelation of the Oct4 and Wnt/β-catenin genes in hepatocellular carcinoma( HCC) cell line HepG2. Methods RTPCR technique was used to detect the expression of Oct4 and β-Catenin in HCC specimens; RNAi was used to knock-down the expression of Oct4 in HepG2, and the change of Wnt/β-catenin related genes were detected by Real time-PCR. Results In HCC specimens, the expression of Oct4 and β-Catenin in tumor and cirrhotic liver tissues were stronger than normal liver tissues. In SiRNA Oct4 HepG2 cells, the expression of Oct4 was downregulated, and β-catenin as well as Wnt10b were in a positive correlation with Oct4, TCF3 was in negative correlation with Oct4. Clone formation and move ability of the HepG2 were downregulated. Conclusions The expression of Oct4 was higher in tumor tissues than in normal liver tissues. Silencing Oct4 by SiRNA-0ct4 in HepG2 resulted in decreased ability of clone formation and cell movement.
4.High metastasis is inhibited in transgenic melanoma cells by endostain gene
Jianda ZHOU ; Yuan HU ; Huiqin XIE ; Jianxiang TAN ; Wenbo LI ; Shaohua WANG ; Jinyan LIU
Journal of Chinese Physician 2009;11(3):308-312
Objective To validate genetic suppression of metastastic capability of highly metastastic melanoma cells by endostatin transfection.Method pcDNA3.1-Endo eukaryotic expression vector contained insulin signal peptide sequence was transfected into highly metastatic mice melanoma cell strain B 16.The expression of endostain was detected by RT-PCR and Western blot experiment,melanoma cells were determined with adhere experiment,in vitro invasion and migration experiment and pulmonary metastasis experiment on C57BL/6 mice.Result Endostatin can obviously inhibit the capability of adherence,in vitro invasion and migration and pulmonary metastasis of melanoma cells.Among them,adhere inhibition ratio was 67.3%,in vitro invasion inhibition ratio was 48.4%,cell migration inhibition ratiowas 52.1%and pulmonary metastasis inhibition ratio was 67.3%.Conclusion Endostatin transfection can obviously inhibit the highly metastatie capability of melanoma cells.
5.Change of Gq-phosphoinositide signaling pathway in left ventricular tissue in rats with chronic heart failure and reverse effect of benazepril on it
Xiaojian SUN ; Shaorong LIU ; Wenbo LIU ; Chuanhuan ZHANG ; Guohui YUAN ; Shehua ZHANG
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To investigate the changes of Gq-phosphoinositide pathway in left ventricular tissue of rats with chronic heart failure in order to assess the role of this signal pathway in the formation of heart failure. METHODS: Male Sprague-Dawle rats were divided into three groups: control, chronic heart failure and benazepril therapy group. Chronic heart failure was induced with adriamycin. Rats in benazepril group received benazepril 10 mg?kg-1?d-1 and adriamycin at the same time. Hemodynamic measurement was carried out after 4 weeks. The expression of G? q/11 protein in left ventricle was detected by Western blotting analysis and activity of phospholipase C was measured by the method of hydrolysis of nuclear substrate. RESULTS: Compared with control group, the ?dp/dtmax in chronic heart failure group significantly decreased, and protein G? q/11 expression, basic and stimulated phospholipase C activity significantly increased (P
6.Formation mechanism of hypercoagulable state after radiofrequency ablation for atrial fibrillation
Huan MENG ; Yuan TAN ; Wenbo DING ; Xianchun HE ; Xiaocui GUO ; Chunzhi TONG ; Junjie KOU
Chinese Journal of cardiovascular Rehabilitation Medicine 2015;24(2):207-210
Pulmonary vein isolation (PVI) is a new ,effective and radical method to cure atrial fibrillation .Within a period after radiofrequency ablation (RFA) ,coagulation system is activated in patients ,then thrombus incident such as cerebral embolism may happen .The present article made a review on its mechanism .
7.Determination of 5-fluorouracil in serum by direct protein precipitation-ultra performance liquid chromatography
Wenbo YUAN ; Yongjuan DING ; Jingjing XU
Journal of China Pharmaceutical University 2015;46(1):81-84
A method based on direct protein precipitation-ultra performance liquid chromatography was established for the determination of trace 5-fluorouracil in serum. The samples were pretreated with 6% perchloric acid as the precipitant and 5-chlorouracil as the internal standard, and the supernatant separated after vortex mixing and high speed centrifugation was used as the test sample. The separation was carried out with an Acquity HSS T3 column and the mobile phase consisting of water and acetonitrile, and the flow rate was 0. 2 mL/min; the injection volume was 5 μL, and the column temperature was 20 °C. The key parameters of this method such as specificity, accuracy, precision and linearity, were validated. Compared with other methods, this method can complete the analysis with significantly reduced time and cost. The established method is useful for therapeutic drug monitoring of 5-fluorouracil plasma concentration and it has been successfully applied in clinical cases.
8.Genetic Characteristics of Coxsackievirus Group A Type 4 Isolated from Patients with Acute Flaccid Paralysis in Shaanxi, China.
Dongyan WANG ; Yi XU ; Yong ZHANG ; Shuangli ZHU ; Yuan SI ; Dongmei YAN ; Hui ZHU ; Qian YANG ; Tianjiao JI ; Wenbo XU
Chinese Journal of Virology 2016;32(2):145-149
We analyzed the genetic characteristics of coxsackievirus A4 (CV-A4) based on the entire VP1 coding region. Samples were isolated from patients with acute flaccid paralysis (AFP) in Shaanxi, China from 2006 to 2010. We wished to ascertain the predominant genotype and the relationship between CV-A4 infection and AFP. Sixty-eight non-polio enteroviruses were inoculated onto RD cells (to increase the virus titer) and molecular typing was undertaken. The entire VP1 coding region was amplified. Percentage of CV-A4 was 10.3% (7/68). Analyses of genetic identify and creation of phylogenetic trees revealed that CV-A4 could be classified into A, B and C genotypes. Seven CV-A4 strains from Shaanxi and other CV-A4 strains from China formed an independent evolution lineage located in group 4 and belonged to the C2 sub-genotype. These data suggested that CV-A4 strains of sub-genotype C2 were the predominant genotypes in China. These strains co-evolved and co-circulated with those from other provinces in China, so continued monitoring of CV-A4 (by clinical and genetic surveillance) should be enhanced.
China
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Enterovirus A, Human
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classification
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genetics
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isolation & purification
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Enterovirus Infections
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virology
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Genotype
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Humans
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Paralysis
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virology
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Phylogeny
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Viral Proteins
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genetics
9.Mechanism of HCV stimulation of human umbilical vein endothelial cells in the pathogenesis of atherosclerosis
Ziyi ZHU ; Yuan LIU ; Wenbo WANG
Journal of Clinical Hepatology 2019;35(6):1313-1317
ObjectiveTo investigate the mechanism of HCV infection in the pathogenesis of atherosclerosis with a model of human umbilical vein endothelial cells (HUVECs) stimulated by HCV in vitro. MethodsHUVECs were stimulated with 1.0 MOI HCVcc for 48 hours. CCK8 assay was used to measure cell proliferation; flow cytometry was used to measure cell apoptosis and cell cycle; wound healing assay and monocyte-endothelial adhesion assay were used to evaluate the influence of HCV on the migration and adhesion of HUVECs; quantitative real-time PCR and Western blot were used to measure the expression of inflammatory factors and endothelial injury factors in HUVECs stimulated by HCV. The two-independent-samples t test was used for comparison between two groups; an analysis of variance was used for comparison between multiple groups, and the LSD-t test was used for further comparison between two groups. ResultsCompared with the control group, the HCV group had no significant changes in the growth, apoptosis, and cell cycle of HUVECs (all P>0.05). HCV stimulation inhibited the migration of HUVECs and enhanced their adhesion ability. Compared with the control group, the HCV group had significant increases in the mRNA and protein expression of the inflammatory factors interleukin-6 and interleukin-1β and the chemokines CXCL10 and monocyte chemotactic protein 1 (mRNA expression: t=-10.155, -12.048, -5.025, and -20.116, all P<0.05; protein expression: F=2541.739, 4806.490, 477.608, and 501.38, all P<0.001). HCV stimulation significantly upregulated the expression of the endothelial injury factors endothelin-1 and vascular endothelial growth factor and the adhesion molecules intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in HUVECs (t=-4.530, -4.497, -7.692, and -7.449, all P<0.05). ConclusionHCV can cause inflammatory changes and dysfunction in endothelial cells and thus affect the development of atherosclerosis.
10.Pathogen isolation and whole-genome sequence analysis of human adenovirus type 55 outbreak on plateaus
Wenbo WANG ; Yuan LIU ; Yifan ZHOU ; Liangqi GU ; Xuelian ZHANG ; Lin ZHANG ; Maomao CHEN ; Xianjun YANG ; Ziying ZOU ; Ping GUO ; Wei QIU ; Xiaobing HU ; Quanshui FAN
Military Medical Sciences 2017;41(6):453-456
Objective To investigate the variation characteristics of adenovirus type 55 (HAdV-B55) gene on plateaus.Methods Throat swabs were collected from HAdV-B55 infected patients and used for virus isolation in HEp-2 cells.The whole-genome sequence was obtained by PCR and sequencing.HAdV-B55 gene sequence was blast with the previously reported virus.Results HAdV-B55 strains were isolated from throat swabs, which were named LS89/Tibet/2016.The whole-genome sequence was obtained and submitted to GenBank with the accession number of KY002683.No large fragment gene recombination was found between this HAdV-B55 strain and previous strains, and the sequence similarity with QS-DLL strain was 99.9%.Conclusion This study provides more information for the evolution patterns of adenovirus 55 and will contribute to the prevention and control of HAdV-B55 infection in the future.