2.Adenovirus construction of expression and its function of connective tissue growth factor.
Ning GONG ; Hua-Bing ZHANG ; Fu-De FANG ; Yong-Sheng CHANG
Acta Academiae Medicinae Sinicae 2011;33(6):649-653
OBJECTIVETo construct and identify a adenovirus vector of the expression of connective tissue growth factor (CTGF) and to explore the role of CTGF in the metabolism of glucose and lipid.
METHODSThe over-expressed plasmid of CTGF was cloned, and then the CTGF sequences were cloned into pAdTrack-CMW vector. The reformed E. coli BJ5183-sensitive bacteria that contain pAdEasy-1 were transformed with lined vector cut by Pme I enzyme. The recombinant adenovirus vector was cut with Pac I enzyme and obtained, then transfected 293A cells to produce virus. Through three times of amplification, the adenovirus infected the primary hepatocytes to determine the infection efficiency and CTGF expression. The mice were starved for several time periods, and then the liver RNA was extracted for real-time PCR to detect the expressions of CTGF under different nutritional conditions.
RESULTSThe adenovirus of CTGF was successfully produced with an infection efficiency of 90%. The expressions of the CTGF were different under different nutritional conditions and showed a coincidence with the expression of peroxisome proliferators-activated receptor gamma coactivator 1 alpha. After the mice were starved for 24h, the expression of CTGF increased by (2.38 +/- 0.51) folds; after the mice were starved for 48 h, the expression of CTGF increased by (2.95 +/- 0.57) folds (P < 0.05).
CONCLUSIONCTGF is speculated to be involved in the metabolism of glucose and lipids.
Adenoviridae ; genetics ; Animals ; Cell Line ; Connective Tissue Growth Factor ; genetics ; Escherichia coli ; genetics ; Genetic Vectors ; Mice ; Mice, Inbred C57BL ; Plasmids ; Transfection
3.Regulatory factors in the articular cartilage repair of knee osteoarthritis
Yu-Qi ZHU ; Zhi-Yao WANG ; Shuai ZHANG ; De-Hua NING
Chinese Journal of Tissue Engineering Research 2017;21(36):5873-5878
BACKGROUND:Knee osteoarthritis is a joint and articular cartilage degenerative disease,and its biological changes mainly include proliferation and apoptosis of chondrocytes.Articular cartilage holds poor regeneration ability,and tissue-engineered cartilage is of great significance for the articular cartilage repair,while cytokines is a major concem for this repair process.OBJECTIVE:To overview the main regulatory factors involved in the articular cartilage repair and chondrocyte apoptosis.METHODS:PubMed and WanFang databases were retrieved for the literature addressing articular cartilage repair and main regulatory factors involved in articular cartilage repair and chondrocyte apoptosis published from 1999 to 2016.Finally 44 eligible articles were included for analysis.RESULTS AND CONCLUSION:Various cytokines in different human tissues are closely related to articular cartilage repair,chondrocyte apoptosis and pathological changes of osteoarthritis,which are involved in chondrocyte damage,degradation of cartilage matrix,synovial degeneration and periostosis.There is an increase in the levels of interleukin 1 β and 6,and tumor necrosis factor α following articular cartilage injury.Thereafter,blocking the expression of these cytokines can protect the articular cartilage from damage.Insulin-like growth factor and transforming growth factor play an important regulatory effect on the chondrocyte proliferation and matrix synthesis.Furthermore,various cytokines regulate the articular cartilage repair and reconstruction via complicated pathways.
4.Domestic and foreign hospital assessment systems and their problems
Chinese Journal of Medical Library and Information Science 2017;26(9):26-32
The aims, indexes, methods, results, data sources, advantages and disadvantages of 12 domestic and foreign hospital assessment systems were analyzed and the problems in current hospital assessment were described in order to provide reference and lessons for perfecting the domestic hospital assessment systems.
5.Clinical Value of Perioperative Plasma D-Dimer in Pediatric Radiofrequency Ablation
xiao-dong, YANG ; min, HUANG ; de-ning, LIAO ; hong, ZHANG ; yuan, GAO ; yang-de, HUA ; jin-kang, LI ; jian-yi, WANG ; jie, SHEN ; yu-juan, HUANG ; xiu-yu, CHEN
Journal of Applied Clinical Pediatrics 1986;0(01):-
Objective To explore the effects of radiofrequency catheter ablation(RFCA)on the blood coaguable states and the clinical value of perioperative plasma D-dimer.Methods The plasma level of D-dimer was assayed by enzyme-linked immunosorbent assay(ELISA)in blood samples of 30 children who were undertaken RFCA.Blood samples were consecutively obtained before cannulating,after electrophysiologic(EP)study,immediately after RFCA,the second day and the seventh day after RFCA.The centrifuged spead was 3 000 r/min,keep it for 10 minutes to obtain the upper plasma,and the crvopreserve.Results The plasma levels of D-dimer was highest at the time point when RFCA was successfully accomplished and restored to preoperative level in the seventh day after RFCA.There were statistically significant difference in the paried values at different time points(Pa
6.Clinical application of low-molecular-weight heparin (Fraxiparine) in rescuing venous crisis of island skin flap.
Bing XIONG ; Hua-de CHEN ; Wen LAI ; Shao-yi ZHENG ; Hui GAO ; Hui-ning BIAN ; Zu-an LIU ; Zhi-feng HUANG ; Li-jun WEI
Journal of Southern Medical University 2011;31(9):1582-1584
OBJECTIVETo evaluate the clinical efficacy of low molecular weight heparin (Fraxiparine) in rescuing venous crisis of island skin flap.
METHODSOf the 73 patients with venous crisis of island skin flap, 47 received subcutaneous injection of low-molecular-weight heparin (group I) and 26 were treated with phlebotomy, local compression and topical application of unfractionated heparin solution gauze (group II).
RESULTSThe flap survival ratio was (88.46∓8.64)% in group I and (38.37∓6.53)% in group II (P<0.001). At 0, 2, and 4 h after injection of low-molecular-weight heparin, the activated partial thromboplastin time (APTT) was obviously delayed (24.28∓6.71, 41.35∓7.64 and 32.34∓6.35, respectively, P<0.01), FXa:C level was significantly decreased (152.4∓30.7, 65.8∓24.4 and 83.4∓18.4, respectively, P<0.01), while FIIa:C level underwent no obvious alterations (155.70∓31.61, 143.20∓24.75, and 143.4∓23.35, respectively, P=NS).
CONCLUSIONFraxiparine has good antithrombotic efficacy in rescuing venous crisis of island skin flap without adverse effect on systemic coagulation.
Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Nadroparin ; therapeutic use ; Surgical Flaps ; adverse effects ; Treatment Outcome ; Young Adult
7.Binding pancreatic duct to mucosa anastomosis.
Shu-you PENG ; Jiang-tao LI ; Li-ping CAO ; Ling-hua ZHU ; De-fei HONG ; Ning LI ; Ying-bin LIU ; Yi-fan WANG ; Yuan-quan YU
Chinese Journal of Surgery 2011;49(9):834-838
OBJECTIVETo study the feasibility of binding pancreatic duct to mucosa anastomosis (BDM)-a complementary procedure to both binding pancreaticojejunostomy and binding pancreaticogastrostomy.
METHODS(1) Animal experimental study:gastrostomy and jejunostomy were performed on six adult New Zealand rabbits. The gastrostomy and jejunostomy shared a same stent (rubber urethral catheter, silicone tube or plastic infusion tube). Both ends of the stent were placed in gastric and enteric cavity. Purse-string suture was performed around the stent before the jejunum and the stomach were brought together for fixation by few stitches. And to observe whether the purse-string suture around a plastic tube, rubber tube or silicon tube inserted into jejunum and/or stomach can prevent leaking out of the jejunal or gastric content to cause peritonitis. (2) Clinically 7 patients were performed with BDM anastomosis. The procedure was consisted of five steps: preparation of the pancreatic stump;preparation of the jejunum; preparation of the fixing sutures between the pancreatic stump and the jejunum; implementation of the anastomosis; lastly, fixation of the jejunum beside the pancreas stump. Post-operative periodic examination of the blood amylase and the amylase in the abdominal drainage. Pancreatic fistula was classified in to two categories: parenchymal fistula (pancreatic cut surface fistula) and anastomotic leakage.
RESULTSAnimal experiment did not show any leakage around the plastic tube or silicon tube inserted into jejunum and(or) stomach. There was no anastomotic leak in all the patients. There was transient increase of amylase in two cases, but the volume of drainage did not exceed 50 ml/d and the recovery of the patients was not affected.
CONCLUSIONSBDM is a simple, safe and easy procedure to perform. It provides to the surgeons with a new option in different situations to achieve the most ideal surgical result.
Anastomosis, Surgical ; methods ; Animals ; Gastric Mucosa ; surgery ; Intestinal Mucosa ; surgery ; Pancreatic Ducts ; surgery ; Pancreaticoduodenectomy ; methods ; Pancreaticojejunostomy ; methods ; Rabbits
8.Killing effect of double suicide genes mediated by retroviral vector on k562 cells.
Yi-Rong JIANG ; Ying-Chang LAI ; Xiao-Lin CHEN ; De-Sheng WAN ; Wan-Ning CHEN ; Miao-Hua QI ; Chun-Sheng LIU ; Xue-Liang CHEN ; Dao-Xin MA
Journal of Experimental Hematology 2007;15(1):47-51
The aim of study was to investigate the killing effect of double suicide gene system mediated by retroviral vector on K562 cells in vivo and ex vivo. CDglyTK gene was transfected into PA317 cells by using lipofectamine. K562 cells were infected with viral supernatant. K562/CDglyTK cells were treated with 5-fluorocytosine (5-FC) and/or ganciclovir (GCV). Mice were randomly divided into three groups: tumor formation, tumor inhibition and tumor therapy. Each mouse was implanted with K562/CDglyTK cells or K562 cells. The results indicated that the killing effect of 5-FC in combination with GCV on K562/CDglyTK was more significant than using 5-FC or GCV alone. In vivo study showed that after being injected subcutaneously with K562 cells and K562/CDglyTK cells, there was not obvious difference in tumor formation rate of mice, 5-FC + GCV could suppress tumor formation of the K562/CDglyTK cells. After being treated with 5-FC and GCV, the median tumor volume of mice implanted with K562/CDglyTK cells decreased obviously, compared with the control group. Their median survival was significantly prolonged. It is concluded that double suicide genes are more effective for killing effect on K562 cells in vivo and in ex vivo. It may be applicable to clinical gene therapy.
Cytosine Deaminase
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genetics
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Flucytosine
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pharmacology
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Ganciclovir
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pharmacology
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Genes, Transgenic, Suicide
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genetics
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Genetic Therapy
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Genetic Vectors
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genetics
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Humans
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K562 Cells
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Protein-Tyrosine Kinases
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genetics
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Receptor Protein-Tyrosine Kinases
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biosynthesis
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genetics
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Recombinant Fusion Proteins
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genetics
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Recombination, Genetic
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Retroviridae
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genetics
9.Impact of different termination modes on atrial fibrillation termination in catheter ablation of persistent atrial fibrillation.
Ping WANG ; Jian-Zeng DONG ; De-Yong LONG ; Man NING ; Ri-Bo TANG ; Rong-Hui YU ; Zeng-Ming XUE ; Cai-Hua SANG ; Chen-Xi JIANG ; Chang-Sheng MA
Chinese Medical Journal 2012;125(11):1877-1883
BACKGROUNDThe optimal endpoint for catheter ablation of persistent atrial fibrillation (AF) remains ambiguous. This study investigated the impact of AF termination as a procedural endpoint and the termination mode on long-term clinical outcome.
METHODSTwo hundred and ninety-three patients who underwent stepwise ablation for persistent AF were categorized into the AF termination by ablation group and into the electrical cardioversion (CV) group. Subgroups were also analyzed based on different termination modes. Follow-up assessment included early recurrence and sinus rhythm (SR) maintenance.
RESULTSDuring initial ablation, 33 patients (11.3%) were directly converted to SR, 166 patients (56.7%) were converted to atrial tachycardia (AT) that subsequently restored SR with further ablation in 98 patients (33.4%), and a total of 162 patients (55.3%) underwent cardioversion due to persistent atrial arrhythmias. Comparison between termination by ablation and termination by cardioversion in patients exhibiting AF or AT revealed that no significant difference was observed in early recurrence (38.2% vs. 43.8%, P = 0.328) and SR maintenance (67.2% vs. 59.8%, P = 0.198) during the (23 ± 7) months follow-up. Even after repeat ablation, the SR maintenance continued to exhibit no statistical difference in above two groups (72.5% vs. 70.4%, P = 0.686). Further analysis of subgroups, however, demonstrated that patients with AF terminated directly to SR experienced better clinical outcomes than other subgroups (P < 0.05). Furthermore, atrial arrhythmias present during ablation have been implicated in prediction of recurrence mode: AF or AT (P < 0.05).
CONCLUSIONSTermination as a procedural endpoint is not associated with favorable long-term SR maintenance in persistent AF. AF methods that convert arrhythmia directly to SR have, however, been linked with improved clinical outcomes, although conversions to AT may not be correlated. Atrial arrhythmias observed during the ablation may be used to predict the recurrence mode.
Adult ; Aged ; Atrial Fibrillation ; physiopathology ; therapy ; Catheter Ablation ; methods ; Electrophysiologic Techniques, Cardiac ; Female ; Humans ; Male ; Middle Aged
10.Establishment of murine allogeneic umbilical cord blood cell transplantation model and study on the mechanism of keratinocyte growth factor enhancing immune reconstitution after transplantation..
Yi WANG ; Guang-Hua CHEN ; De-Pei WU ; Hai-Wen HUANG ; Ying WANG ; Yang XU ; Xiao MA ; Ai-Ning SUN ; Hui-Rong CHANG
Chinese Journal of Hematology 2010;31(2):82-87
OBJECTIVETo explore the impact and mechanism of keratinocyte growth factor (KGF) on immune reconstitution post murine allogeneic umbilical cord blood transplantation (UCBT).
METHODSPerpheral blood (PB) from 19.5-day embryos post-conception (E 19.5 d) mice was used as umbilical cord blood (UCB) graft. Thirty-two BALB/c mice were randomly assigned to 4 groups with 8 mice each in the first cohort UCBT. Mice were infused with PBS (control group) or 1 x 10(6) (group 1A), 2 x 10(6) (group 1B), 3 x 10(6) UCB mononuclear cells (MNCs) (group 1C), respectively. Twenty-four BALB/c mice were randomly assigned to 3 groups with 8 mice each in the second cohort UCBT. Mice were injected with 1 x 10(6) (group 2A), 2 x 10(6) (group 1B) or 3 x 10(6) (UCB) MNCs (group 2C). All mice received platelet transfusion on +8d. Sixteen BALB/c mice were randomly assigned to 2 groups with 8 mice each in the third cohort UCBT. Mice were injected s.c. with KGF (group 3) or PBS (control group) before TBI. All mice were injected with 2 x 10(6) UCB MNCs and were supported with platelet transfusion on +8 d. The survival time, splenic lymphoid cell subsets, sjTREC assay were observed after UCBT.
RESULTSThe 100-day survival of mice were 2, 3 and 3 in group 1A, 1B, 1C and 7, 8, 8 in group 2A, 2B, 2C, respectively. The splenic T, NKT, NK and B cell counts on +35 d were (9.57 +/- 0.74) x 10(6), (0.64 +/- 0.06) x 10(6), (1.43 +/- 0.10) x 10(6) and (19.13 +/- 1.50) x 10(6) in control group, respectively; while were (13.47 +/- 0.74) x 10(6), (0.89 +/- 0.03) x 10(6), (1.79 +/- 0.04) x 10(6) and (20.50 +/- 0.91) x 10(6) in group 3, respectively, being significantly higher than in control group. The sjTREC level was 182.2 +/- 10.7copies per 10(5) cells in control group; while was 224.2 +/- 9.6 copied per 10(5) cells in group 3, being significantly higher than in control group (P = 0.019).
CONCLUSIONSPeripheral blood from E19.5d is rich in hematopoietic stem cells. A murine allogeneic UCBT model with platelet support on +8 d is established. KGF treatment can enhance thymic output and improve T cell immune reconstitution after UCBT.
Animals ; Cord Blood Stem Cell Transplantation ; Fetal Blood ; transplantation ; Fibroblast Growth Factor 7 ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Mice ; Mice, Inbred BALB C