1.Shear stress-regulated vascular endothelial permeability is mediated by Rho GTPases
Ting GAO ; Longjuan XUE ; Xiaoheng LIU
International Journal of Biomedical Engineering 2009;32(3):162-165,176
Dysfunctions of vascular endothelial permeability are related to a number of human diseases such as atherosclerosis, high blood pressure, stroke, inflammation, cancer, diabetes-induced retinopathy, macular edema and so on. Shear stress is an important mechanical force that affects vascular endothelial cells. It plays a particular role in permeability regulation. Rho GTPases is a family of small G proteins which act as cell signal molecules. They are assumed to mediate the regulation of the permeability of vascular endothelial cells. In this paper review is given on how shear stress regulates the permeability of vascular endothehal cells as well as the in-fluence of Rho GTPases on the role in molecular mechanism. It is suggested that shear stress-regulated vascular endothelial permeability is mediated by Rho GTPases.
2.Species identification of freshwater snail Planorbella trivolvis and analysis of its potential distribution
Xiaoheng LI ; Shitong GAO ; Wenbiao GU ; Yi ZHANG ; Yunhai GUO
Chinese Journal of Schistosomiasis Control 2015;(3):268-272
Objective To identify the species classification of an ornamental Planorbidae from a flower market in Shanghai and analyze its potential distribution in China. Methods In August 2013,six freshwater snail specimens were collected from the Wanshang flower market. The species was identified by morphology and molecular biology. An ecological niche model was constructed based on the native geographic presence occurrence data,and projected onto the whole of China to predict the poten?tial distribution. Results Their shell external morphology suggested that the specimens belonged to Planorbella trivolvis(Say 1817)of Planorbidae,which is native in North America. The sequence data of a fragment of the mitochondrial cytochrome c oxi?dase subunit I(COI)confirmed its identification. A total of 2 294 georeferenced occurrence points in North America were car?ried out from the Global Biodiversity Information Facility databases and 614 records with coordinates were used to produce a North American native niche model by a maximum entropy method(Maxent). The projection on China results suggested high probabilities of occurrence mostly in Henan Province and its borderland with nearby provinces. Conclusions P. trivolvis is sim?ilarly with Biomphalaria species from shell morphology. It is the first records of the species in China,and the field dispersal is not clear.
3.Study on distribution of cohere antigen of 12D5 and 21B7 monoclonal antibodies of Angiostrongylus cantonensis and its application of immnodiagnosis
Renli ZHANG ; Muxin CHEN ; Yijie GENG ; Dana HUANG ; Shuang WU ; Shitong GAO ; Xiaoheng LI ; Xingquan ZHU
Chinese Journal of Microbiology and Immunology 2011;31(2):173-177
Objective To detect infection of Angiostrongylus cantonensis and examine effection of treatment to prepare monoclonal antibodies(McAbs). Methods Six-week-old BALB/c mice were imrnunized by the intraperitoneal injection of e/s antigens of Angiostrongylus cantonensis. Fusion of splecn cells from immunized mice with prepared SP2/0-Ag14 myeloma cells was performed in RPMI 1640. Fused cells were suspended in RPMI 1640 containing 1% HAT and 20% fetal calf serum and dispensed into 96-well cell culture plates. The supernatants of clones were screened by ELISA with sera of patients of angiostrongyliasis.Distribution of cohere antigen of 12D5 and 21B7 monoclonal antibodies was analyzed with immunohistochemistry. Two McAbs ( 12D5 and 21B7) were applied to detect the circulating antigen (CAg) in the sera of rats infected with A. cantonensis and angiostrongyliasis patients respectively by double antibody sandwich ELISA.Results 12D5 McAb was identified as IgG1 and 21 B7 McAb was IgM. Western blot result showed two McAbs could used to identified 55 × 103 protein of adult worms of A. cantonensis. Cohere antigen of 12D5 and 21B7 monoclonal antibodies were distributed on intestine surface of A. cantonensis. The detection rates of CAg in the sera of infected rats 100% (48/48), the detection rates of CAg in the sera of angiostrongyliasis patients was 100% (32/32). No cross-reaction to sera of patients with other infection of parasites, such as clonochiasis, fasiolopsiasis, ancylostomiasis, trichinosis, anisakiasis as well as schsitosomiasis, and health srea did not reacted with 12D5 and 21B7 McAbs,and detaction rate of antibody of angiostrongyliasis patients only reached 75% (24/32) with antigen of A. cantonensis. Conclusion Cohere antigen of 12D5 and 21B7monoclonal antibodies were antigens of enteric epithelium. Sandwich ELISA with 12D5 and 21B7 McAbs showed high specificity act as detecting CAg of A. cantonensis in sera of infection animal and patients. It is apparent that Sandwich ELISA with 12D5 and 21 B7 is not only rapid and simple without requirement of special instrument, but also rather sensitive and specific for the detection of current infection with A. cantonensis.
4.Molecular identification of the species of Biomphalaria snails in Shenzhen reservoir based on 16S rDNA sequences
Shitong GAO ; Xiaoheng LI ; Yijie GENG ; Xu XIE ; Shujiang MEI ; Da'na HUANG ;
Chinese Journal of Endemiology 2016;35(9):636-639
Objective To identify the species of Biomphalaria snails collected in Shenzhen reservoir,based on the mitochondrial 16S rDNA sequences.Methods The 16S rDNA fragments were amplified by PCR from the genome DNA of Biomphalaria snails,and inserted in plasmid pMD-18T for sequencing.The sequence of 16S rDNA fragment and its phylogenetic relationships with those of other species of Biomphalaria snails were analyzed with BLAST and MEGA4 software.Results The amplified 16S rDNA fragment of the Biomphalaria snails was about 466 bp in length.As aligned with the corresponding sequences of the related Biomphalaria species,the identity of nucleotides was 99% with 1 isolate of Biomphaltria straminea (B.straminea),98% with 3 isolates of B.kuhniana,95% with 1 isolate of B.intermedia,and 94% with 1 isolate of B.edisoni.Based on the 16S rDNA sequence,the results of phylogenetic analysis with neighbor-joining (NJ) and unweighted pair-group method with arithmetic means (UPGMA) indicated that the snails had close genetic relationships with the B.straminea isolate (Genbank accession NO.AY030213.1) Conclusion The Biomphalaria snails collected in Shenzhen reservoir could be classified as B.straminea based on the characteristics of 16S rDNA sequence.
5.Effects of IL-8 on the tight junction of vascular endothelial cells
Yi YU ; Xiaoheng LIU ; Yang SHEN ; Ting GAO ; Min GAO ; Yunlong MA ; Qiang FU ; Min KANG ; Jianmin ZHAO
International Journal of Biomedical Engineering 2011;34(6):336-339
Objective To investigate the influence of IL-8 on the tight junction of vascular endothelial cells.Methods Immunofluorescence was used to observe the modality and the distribution of three tight junction proteins (occludin,claudin-5 and ZO-1) of the EA.hy926 cells treated with IL-8 under different concentrations and different times.RT-PCR was used to measure the mRNA expression of these three proteins.Results The results demonstrated that IL-8 could change the distribution of occludin,claudin-5 and ZO-1 in EA.hy926 cells,and the mRNA expression of occludin,claudin-5 and ZO-1 decreased with the increase of IL-8 concentration and treated time.Conclusion The effects of IL-8 on the distribution and the expression of occludin,claudin-5 and ZO-1 are dose and time-dependent.
6.Numerical simulation of the distribution of shear stress on the bottom of parallel plate flow chamber under different inlet velocity conditions.
Ye ZENG ; Xiaoheng LIU ; Yi LAI ; Xianliang HUANG ; Bin MAO ; Ting GAO ; Yang SHEN
Journal of Biomedical Engineering 2010;27(4):785-789
The distribution of shear stress on the bottom of the parallel plate flow chamber under different inlet velocities was analyzed by numerical simulation. In the present experimental study, the projection planes of the relative errors at 0.7% level were obtained, and then the efficient region and the actual entrance length were further corrected by introducing the concept of relative error. The results showed that the efficient region of the chamber increased with the direction of length while the inlet velocity was increased, and the actual entrance length was much greater than that of the theoretical entrance length. Therefore, in accordance to the needed range of shear stress in experiment and to the needed efficient region area, the optimum design of the flow chamber is necessary.
Algorithms
;
Blood Flow Velocity
;
Blood Pressure
;
physiology
;
Computer Simulation
;
Humans
;
Models, Cardiovascular
;
Numerical Analysis, Computer-Assisted
;
Pulsatile Flow
;
Rheology
;
Shear Strength
;
Stress, Mechanical
7.Effect on factors of energy metabolism in rats skeletal muscle by cold traditional Chinese medicine.
Shuhong PENG ; Liping HUANG ; Xiaoheng GAO ; Su ZHANG ; Riyue YU ; Hongning LIU ; Ningying XU
China Journal of Chinese Materia Medica 2009;34(23):3064-3067
OBJECTIVETo investigate the effect of six cold traditional Chinese medicine on the energy metabolism factors in rats skeletal muscle.
METHODThe activity of Na(+)-K(+) -ATPase, Ca(2+) -ATPase and succinate dehydrogenase (SDH), the content of muscle glycogen, and the mRNA expression of skeletal muscle uncoupling protein 3 (UCP3) were measured after rats having been administrated with water extracts of Radix Sophorae Flavescentis, Fructus Gardeniae, Cortex Phellodendri, Radix Scutellariae, Rhizoma Coptidis, and Radix Gentianae respectively at the dose of 6.0, 7.0, 8.4, 6.0, 7.0, 4.0 g x kg(-1) for 30 days.
RESULTThe activity of Na(+) -K(+) -ATPase has been depressed significantly and the content of skeletal muscle glycogen has been increased remarkably by six cold traditional Chinese medicine. The decreased tendency has been found on activity of Ca(2+) -ATPase and SDH, only the Radix scutellariaeg group decreased the Ca(2+) -ATPase activity significantly (P < 0.05), the SDH activity was decreased high significantly by Radix scutellariae, Cortex Phellodendri, Radix Gentianae and significantly by Rhizoma Coptidis. The mRNA expression of UCP3 has been decreased high significantly by all five cold traditional Chinese medicine except Cortex Phellodendri group with the decreased tendency of UCP3 mRNA expression.
CONCLUSIONThe cold traditional Chinese medicine has the significant effects on the skeletal muscle energy metabolism by decreasing the utlization of the glucose and the activity of mitochondria SDH to reduce the production of ATP, and depressing the activity of Na(+)-K(+) -ATPase and Ca(2+) -ATPase to cut down the consumption of ATP, by decreasing the mRNA expression of UCP3 to decrease the heat production.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Energy Metabolism ; drug effects ; Male ; Muscle, Skeletal ; drug effects ; metabolism ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley
8.Study on FAK regulation of migration of vascular endothelial cells depending upon focal adhesion proteins.
Min GAO ; Xiaoheng LIU ; Heng SUN ; Hongyi REN ; Lijuan WANG ; Yang SHEN
Journal of Biomedical Engineering 2013;30(3):567-571
Tumor angiogenesis induced by vascular endothelial cells (VECs) migration is a necessary condition for tumor growth and metastasis. The purpose of this study is to investigate the effect of focal adhesion kinase (FAK) inhibitor (50nmol/mL) on the adhesion and migration of endothelial cells(ECs) and the expression of focal adhesion proteins vinculin, talin and paxillin. Scratch wound migration assay was performed to examine the effect of FAK inhibitor with 50nmol/mL on ECs migration at 0, 5, 10, 30, 60 and 120min, respectively. And immunofluorescence analysis was performed to detect the expression of F-actin in ECs treated with FAK inhibitor within 2h. Western blot was carried out to determine the effect of FAK inhibitor on expression of vinculin, talin and paxillin proteins. The results showed that the migration distance and the expression of F-actin in ECs treated with FAK inhibitor decreased significantly compared with that of the controls, and the level of vinculin showed no significant difference with increasing of treated time of FAK inhibitor. However, the talin and paxillin showed an identical decreasing tendency in 5-10min, but slowly going up in 30min and then after subsequently decreasing. The results of this study proved that blocking phosphorylation of FAK could inhibit VECs adhesion and migration by downregulating focal adhesion proteins so that it may inhibit tumor angiogenesis. This may provide a new approach for tumor therapy.
Cell Adhesion
;
Cell Movement
;
physiology
;
Cells, Cultured
;
Endothelial Cells
;
cytology
;
metabolism
;
Focal Adhesion Protein-Tyrosine Kinases
;
antagonists & inhibitors
;
metabolism
;
Focal Adhesions
;
metabolism
;
physiology
;
Humans
;
Neoplasms
;
blood supply
;
Neovascularization, Pathologic
;
Paxillin
;
metabolism
;
Talin
;
metabolism
;
Vinculin
;
metabolism
9.Effect of conditioned medium of vascular endothelial cells on the epithelial-mesenchymal transition of hepatocellular carcinoma cells.
Bowen XU ; Jia HE ; Wenbo GAO ; Guanyue SU ; Xiaoheng LIU ; Yang SHEN
Journal of Biomedical Engineering 2020;37(3):442-449
This study aims to investigate the effect of substances secreted or metabolized by vascular endothelial cells on epithelial-mesenchymal transition (EMT) of hepatocellular carcinoma cells under indirect co-culture condition. Human hepatocellular carcinoma cell line QGY-7703 was cultured , and then was co-cultured with conditioned medium of human umbilical vein endothelial cells (HUVEC). The morphological changes of QGY-7703 cells were observed by inverted phase contrast microscopy. The migration ability of QGY-7703 cells was analyzed by scratch-wound assays. The effect of conditioned medium on the expression and distribution of EMT related proteins was detected by Western blot and immunofluorescence assays, respectively. The results showed that the QGY-7703 cells gradually changed from polygonal to spindle shape, the migration ability promoted significantly, and both the expression and distribution of EMT related marker changed in a time-dependent manner after co-culturing. The results confirm that vascular endothelial cells can induce EMT in hepatocellular carcinoma cells under indirect co-culture condition.
10.Effects of arsenic trioxide on migration, invasion and apoptosis of hepatocellular carcinoma HepG2 cells.
Jia HE ; Bowen XU ; Wenbo GAO ; Guanyue SU ; Hongchi YU ; Yang SHEN ; Xiaoheng LIU
Journal of Biomedical Engineering 2020;37(1):105-111
The article aims to explore the optimal concentration of arsenic trioxide (As O ) on HepG2 of liver cancer cells, and the effect of As O on the migration, invasion and apoptosis of HepG2 cells. In this study, the activity of HepG2 cells treated with 0, 1, 2, 4, 8, 16, 32 μmol/L As O was tested by CCK-8 method, the semi-inhibitory concentration (IC50) was calculated, and the morphological changes of HepG2 cells were observed after the action of As O at IC50 concentration for 12, 24, 48 h. The effect of As O on cell migration and invasion ability was verified by wound healing experiment and Transwell invasion experiment. Western blot and qRT-PCR were used to detect the effects of As O on the gene and protein expression levels related to cell migration, invasion and apoptosis. The results showed that, compared with the control group, the activity of HepG2 cells decreased with the increase of the concentration of As O treatment, showing a dose-dependent effect, and its IC50 was 7.3 μmol/L. After 24 hours' treatment with 8 μmol/L As O , HepG2 cells underwent significant apoptosis, and its migration and invasion abilities were significantly reduced. In addition, the protein expression levels of RhoA, Cdc42, Rac1 and matrix metalloproteinase-9 (MMP-9) were down-regulated, the protein and mRNA expression levels of anti-apoptotic gene Bcl-2 were significantly down-regulated, and the protein and mRNA expression levels of pro-apoptotic genes Bax and Caspase-3 were significantly up-regulated. The above results indicate that certain concentration of As O can inhibit the migration and invasion of hepatocellular carcinoma cells and promote the apoptosis of hepatocellular carcinoma cells.