1.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.
2.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.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.
6.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 .
7.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
8.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.
9.The effect of pulsed electromagnetic fields on the proliferation of immunomagnetically separated human pre-cartilaginous stem cells
Junfang WANG ; Renyun XIA ; Huang FANG ; Anmin CHEN ; Delong YIN ; Hao CHENG
Chinese Journal of Physical Medicine and Rehabilitation 2009;31(5):296-300
Objective To study the biological effects of pulsed electromagnetic fields (PEMFs) on the pro-liferation of immunomagnetically separated human precartilaginous stem cells (PSCs) in vitro. Methods The cells from an aborted fetus's metaphysis were digested using collagenase. The PSCs were isolated by magnetic cell sorting (MACS), then subcultured and amplified. Flow cytometry, immunohistochemistry, immunofluorescence and RT-PCR analysis were performed to identify the purified PSCs. The PSCs were stimulated by PEMFs at 50 Hz frequency and 1 mT intensity. Cell proliferation was measured at different time points using methyl thiazolyl tetrazolium ( MTT), and the cell growth curve was plotted. Flow cytometry was applied to measure the cell cycle and apoptosis. Results The PSCs were successfully cultured. There was fibroblast growth factor receptor-3 (FGFR-3) on their sur-face. Cell proliferation was promoted after 4 and 6 days of PEMF stimulation. The percentage of cells at the S phase was higher than in a control group. Early, late and total rates of apoptosis in the experimental group decreased signifi-cantly. Conclusion PEMFs can enhance the proliferation and inhibit the apoptosis of human PSCs, and it is possi-ble to cultivate the high density human PSCs in vitro.
10.Effects of low dose X-ray irradiation on proliferation and differentiation of mesenchymal stem cells derived from human umbilical cord into adipocytes and osteoblasts
Hao AI ; Hai BAI ; Cunbang WANG ; Jianfeng OU ; Qiaug ZHAO ; Xia HAN ; Xiaoyan HU ; Zhe CHEN
Chinese Journal of Radiological Medicine and Protection 2011;31(3):290-293
Objective To observe the effects of low dose irradiation (LDR) on proliferation,adipogenesis and osteogenic potential of human umbilical cord mesenchymal stem cells (hucMSCs).Methods hucMSCs were isolated from Wharton's jelly tissue of human umbilical cord by modified tissuepiece inoculation,and flow cytometry was used to detect the expression of specific marker in the hucMSCs.The hucMSCs were randomly divided into two groups:irradiation group undergoing irradiation with the doses 50,100,or 200 mGy respectively,and control group without irradiation.MTT method was applied to evaluate the proliferation of the hucMSCs at different time points with various doses irradiation.The third passage hucMSCs were randomly divided into two groups:irradiation group undergoing low dose irradiation of 200 mGy,and control group without irradiation,and then underwent induction by adipocytic and oesteocytic differentiation induction fluids respectively so as to differentiate into adipocytes and osteoblasts.Oil red O staining was used to detect the activity of alkaline phophatase (ALP),and RT-PCR was used to detect the mRNA expression of core binding factor alpha 1 in human osteoblast.Results After 9-12days,fibroblasts began to swim out of the tissue piece with a confluence rate of 80% 2 weeks later.Within 7 days the absorption values of the hucMSCs undergoing different irradiation doses 2,3,4,5,and 6 days later were all significantly higher than those of the control group(F = 159.17,448.81,265.15,183.93,and 181.83 ,all P <0.01),with the proliferation rates of the 100 mGy subgroup being the highest.After being induced liquid,vacuoles were observed in the irradiated group 2 days later.21 days later,the adipogenic rates of irradiated group was significantly higher than that of the control group (t = 28.25,P <0.01).The ALP activity increased in the irradiated group compared with control group (t=16.87,P <0.01) .The expression level of Cbf-α1 mRNA was up-regulated obviously (t = 14.16,P<0.01).Conclusions LDR promotes the proliferation of hucMSCs,and accelerates the hucMSCs' differentiation into adipocytes and osteoblasts.