2.The stable Expression of VEGF_(165) in Mammalian Cells
Journal of Chinese Physician 2001;0(07):-
Objective To investigate the transfection and expression of pcDNA3 1-VEGF 165 plasmid in mammalian cells. Methods pcDNA3 1-VEGF 165 plasmid was transfected into CHO cells by electroporation and the positive clones were screened by G418. The expression of VEGF 165 gene in the transfected CHO cells were detected by RT-PCR, ELISA and western blotting analysis. The biologic activities of VEGF 165 secreted by the transfected CHO cells were measured by Mile's assay and MTT method. Results G418-resistant CHO cell clones were obtained. RT-PCR, ELISA and western blotting analysis showed that there were the transcription and translation of VEGF 165 gene in anti-G418 CHO cells. The mile's assay showed that the expressed VEGF 165 could increase the vessel permeability, and MTT assay proved that it could promote the proliferation of endothelial cells. Conclusion pcDNA3 1-VEGF 165 plasmid could express VEGF 165 with biological activities in mammalian cells.
3.Construction of academic native resource repository platform
Xia WANG ; Hongxin CAO ; Hao CHEN
Chinese Journal of Medical Library and Information Science 2015;(8):55-58
After the importance to construct academic native resource repository platform was elaborated,the native resource repository platform designed and constructed by Library of Second Military Medical University according to the demands of Second Military Medical University was described, which improved the management efficiency of scientific research and academic achievements.
4.Application of needle-knife precut sphincterotomy in treating acute suppurative cholangitis
Zhanxiong XUE ; Wenxing WANG ; Limiao LIN ; Xuanping XIA ; Hao CHEN
Chinese Journal of Postgraduates of Medicine 2006;0(18):-
Objective To evaluate the application of endoscopic needle- knife precut sphincterotomy (PST) in treating acute suppurative cholangitis. Methods After failure of routine papillary intubation during encoscopic retrograde cholangio-pancreatography (ERCP) in papillary incarcerated stone or inflammatory stenosis cases, needle-knife PST was applied to find the lower opening of the common bile duct. After that, routine papillary sphincterotomy or balloon dilation followed. Then net basket for stone extraction and nasobiliary catheter for drainage were made. Results Eleven of the 12 cases′ stones were extracted successfully, the success rate was 91.7%. In the 11 cases, 5 cases′ incarcerated stones dropped into duodenum automatically after sphincterotomy; 9 cases′ stones were extracted successfully in one treatment while 2 cases′ stones were extracted secondarily after stents were implanted; 1 case′s stone could not be extracted and need surgical treatment after nasobiliary catheter drainage because of stenosis of the lower part of the common bile duct. There was no dead case in all the cases. Conclusions Acute suppurative cholangitis patients, who have papillary incarcerated stones or inflammatory stenosis, can receive more efficacious diagnosis and treatment by applying PST when routine endoscopic papillary intubation fails. PST is an important endoscopic treatment for acute calculous suppurative cholangitis
5.Immortalization of human precartilaginous stem cells by transfecting SV40Tag.
Junfang, WANG ; Huang, FANG ; Renyun, XIA ; Anming, CHEN ; Hao, CHENG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(2):231-4
Immortalized human precartilaginous stem cells (IPSCs) were established to provide stable cell resource for the study of the molecular mechanism of gene targeting on the differentiation of PSCs. Plasmid pCMVSV40T/PUR containing simian virus 40 large T antigen gene (SV40Tag) was transfected into human PSCs by using lipofectin transfection. Colonies were isolated by puromycin selection and expanded by multiple passages. Immunohistochemistry, RT-PCR and Southern blotting were used to identify the transfected cells and to detect the expression and integration of SV40Tag in expanded cell lines. The positive colonies were isolated and subcultured, designated immortalized precartilaginous stem cells (IPSCs), which were confirmed as fibroblast growth factor receptor-3 (FGFR-3) positive cells by immunohistochemistry and RT-PCR. SV40Tag cDNA was found in cultured IPSCs of passage 8 by Southern blotting, and the expressions of SV40Tag mRNA and protein were confirmed by RT-PCR. These findings suggested that IPSCs strain with SV40Tag was constructed successfully.
Cartilage/*cytology
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Cell Proliferation
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Cell Transformation, Viral
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Cells, Cultured
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Fetus
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Simian virus 40/*genetics
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Stem Cells/*cytology
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Transfection
6.Principle and advantages of using viscoelastic agent undergo goniosynechialysis during glaucoma and cataract combined surgery
Jun-Xia, ZHANG ; Ying, WANG ; Lin, CHEN ; Li-Juan, HAO
International Eye Science 2014;(9):1677-1679
To investigate the advantage of using the medical sodium hyaluronate gel undergo goniosynechialysis, and to compare the difference of the viscoelastic agent and perfusion fluid by clinical control study.
●METHODS:Totally 103 cases (103 eyes) patients who had angle- closure glaucoma (ACG) with cataract all in Beijing Tongren Hospital affiliated to Capital Medical University, from September 2012 to February 2013. All the patients had primary glaucoma or recurrence ACG with cataract. Their intraocular pressure (lOP) could be controlled in normal value using one or two anti -glaucoma medications. We divided all the patients into two groups by chance. Experimental group had 53 cases (50 eyes), and control group had 50 cases (53 eyes). Two groups all had been performed phacoemulsification combined with goniosynechialysis by the same experiential operator. But experimental group did goniosynechialysis using the medical sodium hyaluronate gel, control group did goniosynechialysis using perfusion fluid. We observed the change of lOP, central anterior chamber depth and anterior chamber structure of two groups in postoperative 1mo.
●RESULTS: SPSS19. 0 statistics software was utilized to analyze the data, This study selected one- side test α =0. 05, adopted paired t - test or two- sample t - test. We obtained that experimental group lOP decreased 12. 42 ± 0. 04mmHg, anterior chamber depth increased 1. 276 ± 0. 201mm; that control group lOP decreased 6. 56 ± 0. 08mmHg, anterior chamber depth increased 0. 852 ± 0. 132mm. And experimental group anterior chamber angle opened more widely than the other group, the comparison had statistical significance (P<0. 05).
●CONCLUSlON: ln phacoemulsification combine with goniosynechialysis,use medical sodium hyaluronate gel do goniosynechialysis can more effectively decrease lOP. This because the medical sodium hyaluronate gel has high glutinousness, it can separate angle of anterior chamber and controlling lOP effectively.
7.Diagnosis of congenital duodenal obstruction in children with high frequency ultrasound combined with gastrointestinal filling
Xingxing DUAN ; Hao LI ; Qingrong XIA ; Jingbo HE ; Wenjuan CHEN
Chinese Journal of Ultrasonography 2016;25(2):146-149
Objective To investigate the diagnostic value of high frequency color Doppler ultrasound combined with gastrointestinal filling in infants with duodenal obstruction . Methods One hundred infants who were suspected with duodenal obstruction underwent both gastrointestinal ultrasound and upper gastrointestinal tract ,put the surgical results as the ?gold standard ,the values of ultrasonic and upper gastrointestinal tract in the diagnosis of pediatric congenital duodenal obstruction were compared . Results The diagnostic accuracy of ultrasound and upper gastrointestinal tract in duodenal obstruction was 100%and 94% respectively ,the difference was statistically significant ( P = 0 .041) . The accuracy rate of obstruction location was 98 .04% and 84 .31% respectively , also was considered to denote statistical significance ( P = 0 .023 ) . The total diagnostic accuracy of congenital disorders caused by duodenal obstruction was 89% and 49% respectively ,and P = 0 .001 implies statistically significant difference . Conclusions There is a high diagnostic accuracy in high frequency ultrasound combined with gastrointestinal filling in children with congenital duodenal obstruction ,and the diagnosis of some diseases is better than that of the upper gastroenterography .
8.Effect of electroacupuncture on abducens nerve injury in Beagle dog
Yi ZHANG ; Xia CHEN ; Lingyun ZHOU ; Xudong WANG ; Hao YU
Tianjin Medical Journal 2015;(2):147-149
Objective To establish Beagle dogs’model of abducens nerves injury and to observe the clinical therapeu?tic effect of electroacupuncture treatment. Methods Twenty-four Beagle dogs were randomly divided into simple crush group (control group) and crush with electrical stimulation group (experimental group). Cisternal segment of the abducens nerve was given a crush injury, then electrodes were implanted to stimulate the abducens nerve and lateral rectus muscle. Distance between the center of the pupil to medial margin of extraocular adjoin was measured from 1 to 12 weeks after opera?tions. Results All procedures used in the study were well tolerated by Beagle dogs. Electrode implantation to stimulate the lateral rectus muscle and the abducens nerve behind of cavemous sinus was successful. There was no statistical significance of the distance between the two groups from 1 to 2 weeks after operations, and the distance was shorter in experimental group than that in control group from 4 to 12 weeks after operations (P<0.01). Conclusion The animal models established to study electroacupuncture treatment of the injured abducens nerves was successful. Electroacupuncture can promote the re?covery of the injured abducens nerves obviously.
9.Outcome and predictive factor analysis of functional mitral regurgitation after aortic valve replacement in patients with severe aortic insufficiency with left ventricular dilation and dysfunction
Yongxin SUN ; Wenjun DING ; Tao HONG ; Hao CHEN ; Limin XIA ; Dong ZHAO ; Chunsheng WANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2013;(6):368-370
Objective Functional mitral regurgitation (FMR) refers to the systolic regurgitation of mitral valve secondary to compromised cardiac function or geometry abnormity with non-organic change of leaflets and ancillary parts of the valve.Severe aortic insufficiency (AI) with left ventricular dilation and dysfunction is clinically a complex heart disease and its postoperative complications and mortality are higher than usual valvular surgery.And such patients are often accompanied by FMR.It is generally acknowledged that FMR may improve after aortic valve replacement(AVR).This study follow up AI patients with left ventricular dilation and dysfunction and preoperative 2 + < FMR≤3 + to evaluate the outcome of FMR after AVR.Preoperative clinical data is assessed by regression analysis.Methods From January 2000 to April 2011 our hospital treated 74 cases of patients with severe aortic regurgitation combined with left ventricular dilation (LVEDds ≥70 mm) and dysfunction (left ventricle ejection fraction,LVEF≤0.35) accompanied by 2 + < FMR≤3 +.Postoperative follow-up was performed.Results The perioperative mortality rate was 8.1%.Average follow-up time was (14.9 ± 7.7) months and follow-up rate of 83.6%.5 patients died during follow-up.6 months after surgery,the average of FMR was (2.64 ± 1.17) (+),compared with preoperative data.LVEDd,LAD,CPMA,all P > 0.05 compared with the preoperative data.LVEF,PAH,both P < 0.05 compared with preoperative data.3.Multiple regression analysis:FMR pre/FMR post ratio is not correlated with age,gender,weight,LVEDd ≥ 75 mm,LVEF≤0.30,hypertension,ventricular arrhythmia and FMR postoperative improvement.However,PAH ≥ 50 mm Hg(1 mm Hg =0.133 kPa),LAD ≥ 50 mm,PAH≥50 mm Hg,CPMA ≥ 15mm,preoperative atrial fibrillation was negatively correlated with FMR postoperative improvement.Conclusion Severe AI with left ventricular dilation and dysfunction is a critical clinical heart disease and its postoperative complications and mortality are high.PAH ≥ 50 mm Hg,LAD ≥ 50 mm,PAH ≥ 50 nun Hg,CPMA ≥ 15mm,preoperative atrial fibrillation was negatively correlated with FMR postoperative improvement.Since patients with 2 + < FMR ≤3 + usually do not improve or even worsen after AVR,those who have these above conditions preoperatively,should be treated on FMR during AVR.
10.Research on effect of neutrophil elastase inhibitor on blood-brain barrier permeability and cerebral edema in TBI rats
Hai SU ; Zulu SU ; Xuenong HE ; Changlong ZHOU ; Hao CHEN ; Xiaohui XIA ; Yi ZHANG
Chongqing Medicine 2014;(26):3458-3460
Objective To investigate the effect of neutrophil elastase (NE) inhibitor on the blood-brain barrier(BBB) permeabili-ty and hydrocephalus in rats with traumatic brain injury (TBI) .Methods 99 SD rats were randomly divided into the control group , the TBI group and the intervention group(dividing into 5 sub-groups:6 ,24 ,48 ,72 ,168 h) .The hydraulic impact model of rats was duplicated .Sivelestat sodium was given in the intervention group .The NE concentration in the brain tissue ,BBB permeability and brain water content were detected in each group and performed the comparative analysis .Results The NE concentration in the brain tissue ,BBB permeability and brain water content at each timepoint in the TBI group and the intervention groups were higher than those in the control group .The NE concentration at 24 ,48 ,72 ,168 h in the intervention group was lower than that in the TBI group .The BBB permeability and the brain water content at 48 ,72 ,168 h in the intervention group were lower than those in the TBI group(P<0 .05) .Conclusion Sivelestat sodium can inhibit the NE release in TBI rat brain tissue ,reduce the BBB permeability and the occurrence of hydrocephalus ,which indicating that sivelestat sodium has the protective effect on TBI secondary lesion in rat .