1.Effects of myogenic induction, differentiation and transplantation of canine umbilical cord blood stem cells on cell-cell junction
Jun WAN ; Ju MEI ; Jinben MA ; Nan MA ; Genfa SHAN
Chinese Journal of Tissue Engineering Research 2009;13(36):7108-7112
BACKGROUND: Umbilical cord blood-mesenchymal stem cells (UCB-MSCs) following differentiation into cardiomyocytes were transplanted into ischemic myocardium. The transplanted cells can build connection with host cells and repair the infarct myocardium. OBJECTIVE: To detect the cell-cell junction after transplantation of the cardiac-like cell derived from the canine umbilical cord blood stem cells. DESIGN, TIME AND SETTING: A randomized controlled animal study was performed from July 2006 to October 2007 at the Animal Experimental Center of Xinhua Hospital Affiliated to School of Medicine, Shanghai Jiao Tong University. MATERIALS: A total of 2 full-term pregnant canines were used for isolation of UCB-MSCs. A total of 36 adult mongrel canines were divided into cell transplantation group and model control group (n=18) according to the rule of random digits table. METHODS: The MSCs at passage 4 were transfected by Laz-Z. After 3-day culture, MSCs were induced by 10 μmol/L 5-azacytidine (5-aza). The canine models of myocardium infarction were established following 3 weeks of culture. 2 mL (1 ×107)MSCs were transplanted into dogs with acute myocardium infarction by coronary artery infusion and local injection in cell transplantation group. An equal volume of saline was used in the model control group. The specimens were harvested and detected at 2, 4 and 8 weeks, respectively. Cell junction was determined using immunohistochemistry. MAIN OUTCOME MEASURES: The following parameters were measured: gene trensfection, myogenic induction and differentiation results of UCB-MSCs; junction of transplanted cells and host cardiomyocytes. RESULTS: Following 72 hours of transfaction, mass of cells expressed LacZ gene, synthetized galactosidase, and stained blue using X-gal staining. Following 3 weeks of 5-aza induction, the antigen a-Actin, Desmin and Connexin43 were all been positively expressed, but before induction they were all negative. From the myocardial section of 8 weeks after transplantation, the junction was formed between the transplanted cells and the host myocardium as formed between the transplanted cells. In the junction, green-fluorescence positive expression of cadherin and connexin43 could be seen. However, in the model control group, only cadherin and connexin43 expressed positively, but the transplanted UCB-MSCs with red fluorescence could not been observed. CONCLUSION: The UCB-MSCs is able to differentiate into cardiac-like cell in vitro and form cell-cell junction in vivo to communicate with surrounding cells.
2.Effect of umbilical cord blood mesenchymal stem cell transplantation on remaining myocardial tissues of dogs with acute myocardial infarction
Nan MA ; Hong ZHONG ; Dehai CHEN ; Yu JIN ; Genfa SHAN
Chinese Journal of Tissue Engineering Research 2007;11(46):9404-9407
BACKGROUND: Cell apoptosis and ventricle reconstitution following myocardial infarction are of mutual cause-effect, and they cause vicious cycle. How to reduce the apoptosis events following myocardial infarction is one of keys to saving heart function.OBJECTIVE: To observe the effect of umbilical cord blood mesenchymal stem cell (UCBSMC) transplantation on remaining myocardial tissue of dogs with acute myocardial infarction.DESIGN: A randomized controlled observation.SETTING: Department of Cardiothoracic Surgery, Xinhua Hospital.MATERIALS: This study was carried out in the Central Laboratory of Xinhua Hospital from October 2005 to May 2007.Thirty-six adult hybrid dogs, male and female in half, were provided by the Animal Experimental Center of Xinhua Hospital.METHODS: Thirty-six dogs were divided into cell transplantation group and control group, with 18 dogs in each according to table of random digit. Mesenchymal stem cells were isolated from the umbilical cord blood of full-term pregnant hybrid dogs, cultured and amplified. Then, they were labeled with Laz gene, in vitro induced with 5-azacytidine, and transplanted into the dogs with acute myocardial infarction in the cell transplantation group. Rats in the control group were injected with the same amount of normal saline. Each dog was euthanized by anesthesia for harvesting myocardial specimen 1,4 and 8 weeks after transplantation.MAIN OUTCOME MEASURES: ① Remaining and apoptosis index detected by TUNEL method. ② Myocardial cell volume and histomorphology detected by confocal microscopy. ③ Histological change of myocardial collagen network detected by haematoxylin-basic fuchsin-picric acid staining.RESULTS: Thirty-six involved experimental dogs all entered the stage of final analysis. ①The apoptosis index in the cell transplantation group was significantly lower than that in the control group 1, 4 and 8 weeks after cell transplantation (P <0.05). ② Myocardial cell volume in the cell transplantation group 1, 4 and 8 weeks after cell transplantation was significantly larger than that in the control group (P < 0.05). ③ Collagen fiber in the myocardial tissue of dogs in the cell transplantation group was arranged in order and regularly, and in contrast that in the control group was not, and fibers in the control group fused partially.CONCLUSION: UCBSMC transplantation reduces the apoptosis of myocardial cells, promotes the hypertrophy of remaining myocardial cells, regulates myocardial collagen network and improves heart function.
3.Imaging anatomical study of kidney and colon position changes between the prone and low-arch oblique supine positions
Liang GUO ; Nan ZHANG ; Xinheng ZHANG ; Yaohui MA ; Zhongjie SHAN
Chinese Journal of Urology 2015;(6):433-435
Objective To provide imaging anatomy basis for percutaneous nephrolithotomy ( PCNL) by measuring relative displacement and changes in anatomical position of kidney and colon under the prone and low-oblique supine positions.Methods Forty-six patients scheduled for PCNL underwent 64-slice spiral CT scan under the prone and low-arch oblique supine position before the PCNL.The horizontal distance of kidney and colon,the distance from colon and analog puncturing line,the distance between the kidney and colon were measured and compared between the 2 positions.Results The distance from colon and analog puncturing line under the low-oblique supine and prone positions were as follows,the left (26.56 ±15.36) mm versus (12.25 ±13.16) mm (t=3.527,P<0.05),the right (25.85 ±14.26) mm versus (13.57 ± 12.53) mm (t=3.234,P<0.05).The differences of the rest distances between the 2 positions were not significant ( P>0.05).Conclusions The distance between colon and analog puncturing line increases in the low-arch oblique supine position,because the colon shifts to the ventral.The PCNL in low-arch oblique supine position may reduce the incidence of colon injury,and improve surgical safety.
4.N-myc downstream regulated gene 2 promotes apoptosis of bladder cancer cells through signal transducer and activator of transcription 3 signaling pathway
Yang ZHAO ; Yaohui MA ; Xiaolei ZHAI ; Nan ZHANG ; Zhongjie SHAN
Chinese Journal of Urology 2017;38(7):537-541
Objective To investigate the effect of N-myc downstream regulated gene 2 (NDRG2) on the apoptosis of bladder cancer cells by regulating the signal transducer and activator of transcription 3(STAT3) signaling pathway.Methods The expression level of NDRG2 in human bladder cancer cell BIU-87 and immortalized cell SV-HUC-1 was detected by Western blot.NDRG2 over expression vector and empty vector control (pcDNA3.1),siRNA-NDRG2,siRNA control were transfected into BIU-87 cells.After transfected 48 h,the expression level of NDRG2,Cleaved caspase 3,STAT3,p-STAT3,JAK2,p-JAK2 were detected by Western blot,the cell proliferation and apoptosis were measured by MTT and flow cytometry.After adding inhibitor AG490 of 45 μmol/L in cultured BIU-87 cells,MTT assay was used to detect cell proliferation and flow cytometry was used to detect the cell apoptosis,Western blot to detect the expression level of Cleaved caspase 3,STAT3,p-STAT3,JAK2,p-JAK2.Results The expression level of NDRG2 in bladder cancer cells was higher than that in bladder epithelial cells.The cell survival rate of pcDNA3.1/NDRG2 group was lower than that of pcDNA3.1 group,the difference was statistically significant (P < 0.01).The cell survival rate of siRNA-NDRG2 group was higher than that of siRNA control group (P< 0.01).The apoptosis rate of pcDNA3.1/NDRG2 group was higher than that of pcDNA3.1 group (P < 0.01).The apoptosis rate of NDRG2 siRNA group was lower than that of siRNA control group (P < 0.01).The level of p-STAT3 and p-JAK2 in pcDNA3.1/NDRG2 group was lower than that in pcDNA3.1 group (P< 0.01).The survival rate and apoptosis rate of BIU-87 cells cultured with AG490 were in agreement with the trend of pcDNA3.1/NDRG2 after transfection.Conclusions NDRG2 could promote the apoptosis of bladder cancer cells,and its mechanism may be related to STAT3 signaling pathway.
5.Contents Determination of Citric Acid in Fentanyl Citrate Raw Materials and Its Injection by Ion Chroma-tography
Xun MA ; Panpan LI ; Guixia LIU ; Yanping ZONG ; Wei SUN ; Guangzhi SHAN ; Hua CHEN ; Nan NAN
China Pharmacy 2017;28(18):2536-2538
OBJECTIVE:To determine the contents of citric acid in fentanyl citrate raw materials and its injection by ion chro-matography. METHODS:The determination was performed on Thermo Dionex IonPacTM AS11-HC column with mobile phase con-sisted of potassium hydroxide (gradient elution) at the flow rate of 1.0 mL/min. The column temperature was 35 ℃,and sample size was 20 μL. The detector was suppressed conductivity detector. RESULTS:The linear range of citric acid were 0.1157-74.05 μg/mL(r=0.9995). The limit of quantitation was 0.1150 μg/mL,and the limit of detection was 0.0400 μg/mL;RSDs of preci-sion,stability and reproducibility tests were all lower than 2.0%;the average recoveries were 99.6%-101.5%(RSD=0.68%,n=9). CONCLUSIONS:The method is environmentally-friendly and simple with good accuracy and precision,and suitable for the contents determination of citric acid in fentanyl citrate raw materials and injection.
6.Effects of umbilical cord blood endothelial progenitor cell transplantation on angiogenesis following myocardial infarction
Bo LI ; Hongguang ZHAO ; Hong ZHONG ; Ruijun LIU ; Nan MA ; Genfa SHAN ; Ju MEI ; Fuxian ZHANG ; Guoqing LI
Chinese Journal of Tissue Engineering Research 2009;13(27):5375-5379
BACKGROUND: Endothelial progenitor calls are the cells that can form new blood vessels in the way of angiogenesis in the body,which updates the conventional theory of angiogenesis, vascular damage and repair after birth and provides new ideas for research and treatment of ischemic diseases.OBJECTIVE: To investigate the effects of dog umbilical cord blood endothelial progenitor call (UCB-EPC) transplantation on angiogenesis after myocardial infarction.DESIGN, TIME AND SETTING: An in vivo cytological experiment was performed at the Laboratory Center of Xinhua Hospital between May 2006 and March 2007.MATERIALS: One full-term pregnant hybrid dog was included for preparation of UCB-EPCs. Thirty-six adult dogs were randomly divided into a cell transplantation group (n = 18) and a model control group (n = 18).METHODS: Acute myocardial infarction model was established in each group by ligation of antedor descending coronary artery.In the cell transplantation group, 2 mL physiological saline containing 5×10<'6> BrdU-labeled EPCs was injected into the coronary artery, while in the model control group, simple physiological saline of the same amount was given. At 1,4, and 8 weeks after transplantation, dogs were sacrificed for harvesting myocardial tissue.MAIN OUTCOME MEASURES: Myocardial infarction was confirmed by hematoxylin-eosin staining. Myocardial angiogenesis was observed by BrdU immunohistochemical staining. The number of infarcted myocardial vessels was calculated by yon Willebrand (vW) factor staining.RESULTS: There was plenty of scar tissue, flbroblasts, and small vessels in the myocardial infarction region. In the cell transplantation group, brown yellow particles (BrdU-positive expression) appeared in some nuclei in small vessels from infarcted myocardium. Newly formed vessels were not found in the model control group. In the cell transplantation group, brown yellow particles (vW factor-positive expression) appeared in the cytoplasm of the vascular endothelial cells in the myocardial ischemia and infarction regions, vW factors were not expressed in the model control group. At 1, 4, and 8 weeks after myocardial infarction,there was no significant difference in vessel counts no matter in myocardial ischemia region or in myocardial infarction region between the call transplantation and model control groups.CONCLUSION: EPCs derived from UCB of pregnant dog can participate in the formation of blood vessels but can not promote angiogenesis after acute myocardial infarction.
7.Inserting ureterovesical reimplantation by means of laparoscopy associated with six-stitch suture
Nan ZHANG ; Kun CHEN ; Liang GUO ; Yaohui MA ; Lei GE ; Chaohui HAO ; Qianhe HAN ; Jianting HU ; Zhongjie SHAN
China Journal of Endoscopy 2017;23(3):94-98
Objective To discuss the safety and efficacy of inserting ureterovesical reimplantation by means of laparoscopy associated with six-stitch suture.Methods There was an retrospective analysis on operation videos and clinical data for 16 participants of inserting ureterovesical reimplantation by means of laparoscopy associated with six-stitch suture with the period from March in 2012 to September in 2015. And these were statistically analyzed including the operation time, intraoperative bleeding volume, postoperative drainage volume, removal time of drainage tube, admission time after operation and the incidence of postoperative complications of vesicoureteric reflux and stenosis.Results The operations of 16 participants were completed successfully without converting to open surgery. The operation time was 60 ~ 125 min (Mean time: 85 min); intraoperative bleeding volume was 20 ~ 50 ml (Mean volume: 32 ml); postoperative drainage volume was 60 ~ 400 ml (Mean volume: 106 ml); removal time of drainage tube was 3 ~ 6 d (Mean time: 4.2 d) and admission time after operation was 7 ~ 10 d (Mean time: 8.5 d). There was the follow-up with 6 ~ 18 months (Mean time: 12 months) for participants. No anastomotic stenosis was present. In addition, one participant was suffered from mild vesicoureteric relfux. And there was no aggravation during 18 months.Conclusions The inserting ureterovesical reimplantation by means of laparoscopy associated with six-stitch suture was safe and effective. It was found that the operation time was significantly shortened and the incidence of postoperative complications of vesicoureteric relfux and anastomotic stenosis was not increased. By contrast, the six-stitch suture could reduce the incidence of anastomotic stenosis.
8.Association between gene polymorphism of neuropeptide Y promoter and cerebral stroke subtypes according to TOAST criteria
Si-Shan GAO ; Lan TAN ; Nan-Nan YU ; Jin-Tai YU ; Jing-Hui SONG ; Teng MA ; Nai-Dong WANG
Chinese Journal of Neuromedicine 2010;09(10):1037-1041
Objective To explore the relationship between gene polymorphism of neuropeptide Y (NPY) promoter and cerebral stroke subtypes according to TOAST (Trail of ORG 10172 in Acute Stroke Treatment) criteria in Chinese Han population. Methods The gene polymorphisms at position of-399T/C, -883Tgins/del and -602G/T in NPY promoter were detected by PCR method and gene sequencing in 190 cases of large-artery atherosclerosis stroke (LAA), 260 cases of small-artery occlusion (SAO), 60 cases of cardioembolism stroke (CE), 29 cases of stroke of other demonstrated etiology (ODE),10 cases of stroke of other undemonstrated etiology (UE) and 423 healthy control subjects. The PCR products were directly sequenced. Multivariate logistic regression was performed to analyze the relationship between gene polymorphism of NPY promoter and cerebral stroke subtypes according to TOAST by removing the confounding variables. Results Significant differences in the frequency of genotype CC and allele C at position of-399T/C were noted between the patients with SAO and controls (P=0.046, P=0.010). Compared with the control group, patients with LAA and SAO were more likely having high level of uric acid, hypertension, diabetes mellitus and heard disease (P<0.05). No statistic differences in the frequency of genotype DD and allele D at position of-883Tgins/del were noted between patients with SAO and controls (P=0.0605, P=0.155). Gene polymorphisms of-399T/C,-883Tgins/del and -602G/T did not associate with an increased risk of having LAA, CE, ODE and UE.Conclusions The gene polymorphisms of promoter in position of-399T/C gene maybe associate with the happening of SAO; allele C at the position of-399T/C may raise the risk of the disease. There is no relationship between the gene polymorphisms of promoter at position of-399T/C, -883Tgins/del, -602G/T and the patients with LAA, CE, ODE and UE. High level of uric acid, hypertension, diabetes mellitus and heard disease history are the risk factors of LAA and SAO.
9.Improved weighted cross-correlation coefficient with a new kernel and its application in predicting T cell epitopes.
Jing HUANG ; Jian-hua MA ; Nan LIU ; Shan-shan QIAN
Journal of Southern Medical University 2010;30(10):2252-2255
We designed a weighted cross-correlation coefficient considering the "anchor" of the T cell epitopes, and used an evolutionary algorithm to search for an optimal weight vector. A SVM model with this new peptide similarity kernel was evaluated on a T-cell data set. The results demonstrated a good performance of this method.
Algorithms
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Artificial Intelligence
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Epitopes, T-Lymphocyte
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Models, Statistical
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Peptides
10.Protective effect of Chinese drug huoxuehuayu decoction on macula after phacoemulsification
Jing, LIN ; Gui-Qiu, ZHAO ; Yan, MA ; Qing, WANG ; Shan-Shan, YANG ; Li-Ting, HU ; Cheng-Ye, CHE ; Nan, JIANG ; Jing-Fen, ZHANG
International Eye Science 2011;11(3):381-384
AIM: To investigate the protective effect of huoxuehuayu decoction on macula after phacoemulsification. Into A, B groups. The 80 eyes of A group were treated by conventional phacoemulsification; the patients (60 eyes) of B group were given huoxuehuayu decoction orally for two courses after phacoemulsification. Best-corrected visual acuity(BCVA), corneal and aqueous conditions ,thickness of macular central fovea and changes of macular retinal tissue in A, B groups were observed before surgery, 1 day; 1 week,2,4,6,8 weeks and 3 months after surgery. Was significantly higher than that of group A. One week after surgery the ratio of mild aqueous flare in group B was significantly lower than that of group A. The thickness of central fixation was significantly increased in both groups 1 week, 2, 4, 6, 8 weeks and 3 months after phacoemulsification; the difference between 2 to 8 weeks after surgery and pre-operation showed statistical significance in both groups. 11 eyes in A group had macular edema during 2 to 6 weeks after surgery, including 9 eyes with fovea thickened and 2 eyes with cystoid macular edema, and seven eyes' edema disappeared in 3 months. 2 eyes in B group had macular edema, including 1 eye fovea thickened and 1 eye cystoid macular edema, during 4 to 6 weeks after surgery, and the two eyes' edema disappeared 3 months after surgery. The fovea thickness in group B during 2 to 8 weeks after surgery was statistically lower than group A. Phacoemulsification.