1.Experimental study of cartilage defect repair using bone mesenchymal stem cells transfer of BMP-2 and TGF-β3 gene mediated by adenovirus vector combining with deminerized bone matrix in pig
Xin WANG ; Yanlin LI ; Yaofeng JIN ; Jianming CHEN ; Huijian WANG ; Chuan HE ; Shuhai CAO ; Fengkai ZHAO
The Journal of Practical Medicine 2014;(18):2880-2882
Objective To explore the repair result of full-thickness cartilage defects in diannan small-ear pig by bone mesenchymal stem cells (BMSCs) transferred with both transforming growth factor-β3(TGF-β3) and bone morphogenetic protein-2(BMP-2) gene mediated by adenovirus vector and combined with deminerized bone matrix (DBM). Methods 32 full-thickness defects from 16 knees of 8 pigs were randomly divided into 4 groups in the experiments. In group A, the animals′ lateral femoral condyle of right knee joint was repaired with DBM and BMSC infected with both Ad-TGF-β3 and Ad-BMP-2. In group B, the medial femoral condyle of right knee joint was repaired with DBM and BMSC without infection. In group C, the lateral femoral condyle of left knee joint was repaired with DBM. And the group D is control group. Morphology and histology were observed 2, 4, 8 and 12 weeks after operation. Results 12 weeks after operation, the whole defects were repaired in group A, HE staining showed typical cartilaginous structure in the repaired area. In group D, defects were not repaired but filled with fibrous tissue. The O′driscoll scores were 15.65 ± 0.11 (group A), 11.33 ± 0.22 (group B), 6.13 ± 0.15 (group C) and 5.08 ± 0.15 (group D). There was significant difference among the groups (P < 0.05). Conclusions The new type of tissue engineering scaffold that DBM combined with BMSCs transfected with both Ad-BMP-2 and Ad-TGF-β3 could induce cartilage regeneration and repair the defects.
2.Detection of apoptosis of Th1 and Th2 cells in C57BL/6 mice chronically infected with Schistosoma japonicum
Xinyu XU ; Jiaqing ZHAO ; Ying CHI ; Lei HE ; Xiaoyun WEN ; Chuan SU
Chinese Journal of Schistosomiasis Control 1989;0(01):-
Objective To observe the apoptosis of Th1 and Th2 cells in C57BL/6 mice chronically infected with Schistosoma japonicum.Methods The apoptotic Th1 and Th2 cells in spleen and lymph node from C57BL/6 mice infected with Schistosoma japonicum for 13 weeks were examined by three-color and indirect flow cytometery with staining surface molecule and intracellular cytokines.Results Compared with the normal mice,the proportion of apoptotic Th1 and Th2 cells of 13-week post-infection was significantly high,and the apoptotic Th1 cells increased more than apoptotic Th2 cells in the infected C57BL/6 mice,and the Th1 cells were more susceptible to apoptosis than Th2 cells.Conclusions Unequal susceptibility to apoptosis in Th1 and Th2 cells may be one of the reasons leading to Th2 polarization on mice chronically infected with Schistosoma japonicum,which provides the new proof of Th polarization.
3.The effects and molecular mechanisms of simvastatin on liver fibrosis in rats with non-alcoholic fatty liver disease
Wei WANG ; Caiyan ZHAO ; Yadong WANG ; Xian HE ; Chuan SHEN ; Wei CAO ; Dianxing SUN
Chinese Journal of Digestion 2011;31(11):739-743
ObjectiveTo investigate the effects and molecular mechanism of simvastatin in liver fibrosis model of non-alcoholic fatty liver disease (NAFLD) in vivo and hepatic stellate cell in vitro.MethodsFirstly,the rat liver fibrosis model of NAFLD was established by high-fat diet administration and intervened with simvastatin.The expression of endothelial nitric oxide synthase (eNOS),inducible nitric oxide synthase (iNOS) and Collagen Ⅰ at mRNA and protein level were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.Secondly,quiescent phenotype of LX-2 cell line was induced by promoting adipocyte differentiation medium in vitro,and then the quiescent phenotype of LX-2 cell were treated with transforming growth factor β1(TGF-β1),Nitroso-L-arginine methyl ester (L-NAME) which was NOS inhibitor,simvastatin,TGFβ1 with simvastatin,and L-NAME with simvastatin separately.The changes of eNOS,iNOS,αsmooth muscle actin (α-SMA) and Collagen Ⅰexpressions at mRNA and protein level were determined by RT-PCR and Western blot.ResultsAs modeling time extended,the expressions of eNOS in rat's liver tissue of model group at mRNA and protein level decreased gradually,however the expression of iNOS and Collagen Ⅰ at mRNA and protein level increased gradually,compared with normal control group and the differences were statistically significant (P <0.05 and 0.01).By 24weeks,the expressions of eNOS in rat's liver tissue of simvastatin group at mRNA and protein level were increased,the expression of iNOS at mRNA and protein level were decreased and the expression of Collagen Ⅰ at mRNA and protein level were decreased (all P <0.05).The expression of eNOS in rat's liver tissue of model group negatively correlated with the expression of Collagen Ⅰ at mRNA and protein level (all P <0.01).The expression of iNOS positively correlated with that of Collagen Ⅰ at mRNA and protein level (all P <0.01).In LX-2 cell culture,L-NAME inhibited the activation of LX-2,reduced eNOS and iNOS expression and increased α-SMA and CollagenⅠexpression,consistent with the role of TGF-β1.Simvastatin could directly increase the eNOS expression both in quiescent and activated LX-2 cells,decrease iNOS expression,maintain quiescent phenotype and inhibit its activation.ConclusionsSimvastatin ameliorated the genesis and progression of liver fibrosis by increasing eNOS expression in LX-2 cells and reducing iNOS,α-SMA and Collagen Ⅰ expression.
4.Diagnostic Value of Nebulized Bronchodilator Test with Tidal Breathing Analysis for Infants with Asthma
shuo, LI ; chuan-he, LIU ; xin, SONG ; jing, ZHAO ; yu-zhi, CHEN
Journal of Applied Clinical Pediatrics 2006;0(19):-
Objective To evaluate the effect of nebulized bronchodilator on the change of tidal breathing in infants with asthma.This may provide some objective evidence for clinical diagnosis.Methods One hundred and five infants with asthma and 26 cases with pneumonia were involved for the study.Tidal breathing flow-volume was obtained before and 10-15 minutes after nebulized bronchodilator was given.Accor-ding to the basic lung function results,59 cases as a mild group [the ratio of time taken to reach peak expiratory flow to total expiratory time(TPTEF/Te) ≥15%],and 46 cases of the infant asthma as a severe group(TPTEF/Te0.05).Moreover,in severe group,expect for RR and expiratory time(Te),the other indices significantly increased after administration of nebulized bronchodilator(P
5.Factors as predictors for thoracic and thoracolumbar/lumbar structural curves in adolescent idiopathic scoliosis.
Zi-qiang CHEN ; Yong-fei ZHAO ; Shi-sheng HE ; Chuan-feng WANG ; Jing-tao ZHANG ; Ying-chuan ZHAO ; Chang-wei YANG ; Ming LI
Chinese Medical Journal 2012;125(8):1439-1442
BACKGROUNDRecent studies have demonstrated that the Lenke system is relatively efficient and consistent in classifying scoliosis curves. Basically, fusion should include the main curve and the structural minor curve. The criteria for defining the structural minor curve were established to help guide these decision-making process. The present study was designed to investigate predictors of the structural curve, and see whether it was possible to prevent the formation of the structural curve by interfering with influencing factors to decrease the fusion level.
METHODSAge, gender, Cobb angle, Perdriolle rotation, Risser sign and the number of vertebrae included in the curve, brace treatment, and curve location were recorded in 145 idiopathic scoliosis patients from July 2001 to January 2007. The patients were divided into two groups: structural and non-structural groups. Demographics and baseline characteristics were compared between the two groups as an initial screen. Logistic regression was used to analyze factors affecting the minor curve to become the structural curve.
RESULTSCompared with the non-structural group, the structural group had a higher Cobb angle ((51.34 ± 13.61)° vs. (34.20 ± 7.21)°, P < 0.001), bending angle ((33.94 ± 9.92)° vs. (8.46 ± 5.56)°, P < 0.001) and curve rotation ((23.25 ± 12.86)° vs. (14.21 ± 8.55)°, P < 0.001), and lower flexibility ((33.48 ± 12.53)% vs. (75.50 ± 15.52)%, P < 0.001). There was no significant difference in other parameters between the two groups. The results of the Logistic regression analysis showed that the Cobb angle (OR: 9.921, P < 0.001) and curve location (OR: 4.119, P = 0.016) were significant predictors of structural curve in adolescent idiopathic scoliosis. Every 10° change of Cobb angle increased the possibility of turning the minor curve into the structural curve by 10-fold. And thoracic curve showed, on the average, the possibility of becoming the structural curve about 4-fold more often than did the thoracolumbar/lumbar curve.
CONCLUSIONSCurve severity and curve location affect the minor curve's structural features in adolescent idiopathic scoliosis.
Adolescent ; Female ; Humans ; Logistic Models ; Lumbar Vertebrae ; pathology ; Male ; Scoliosis ; pathology ; Thoracic Vertebrae ; pathology
6.Endoscopic papillary balloon dilatation vs. endoscopic sphincteropapillotomy for common bile duct stones: a meta analysis.
Liang HE ; Xiao-ping GENG ; Hong-chuan ZHAO ; Da-chen ZHOU ; Fu-bao LIU ; Yi-jun ZHAO ; Guo-bin WANG ; Zhi-gong ZHANG ; Fan HUANG
Chinese Journal of Surgery 2013;51(6):556-561
OBJECTIVETo evaluate the safety and efficacy between endoscopic papillary balloon dilatation (EPBD) and endoscopic sphincteropapillotomy ( EST) for common bile duct stones using meta-analysis method.
METHODSRandomizd controlled trials comparing EPBD with EST for common bile duct stones and published from January 1990 to July 2012 were recruited. This meta-analysis was conducted to estimate short-term and long-term complications. Fixed random effect model or random effect model was established to analyze the data.
RESULTSTwelve randomizd controlled trials were included in this analysis. These studies included 1865 patients, 925 of them were treated with EPBD and 940 were treated with EST. The analysis of basic characteristics of these included studies showed that: compared to EST, patients in the EPBD group were younger (OR = -1.16, 95% CI: -1.49 to -0.84, P = 0.00), while in two groups, there were no significant difference (P > 0.05) in gender proportion, average size of stones, number of gallstones, previous cholecystectomy, the number of merged duodenal diverticulum, common bile duct diameter, the total follow-up time. Also, compared to EST, the overall stone clearance in the EPBD group was lower (OR = 0.64, 95% CI: 0.42 to 0.96, P = 0.03), pancreatitis incidence was higher (OR = 2.67, 95% CI: 1.61 to 4.43, P = 0.00), incidence of bleeding (OR = 0.12, 95% CI: 0.04 to 0.34, P = 0.00), acute cholecystitis (OR= 0.39, 95% CI: 0.18 to 0.84, P = 0.02), total long-term complication rate (OR = 0.53, 95% CI: 0.36 to 0.77, P = 0.01), stone recurrence rate more than a year were lower (OR= 0.48, 95% CI: 0.26 to 0.90, P = 0.02). While in two groups, there were no significant difference (P > 0.05) in the stone removal on 1 '' attempt, the total near-term complications and acute cholangitis.
CONCLUSIONSOn the basis of lower rates of bleeding, EPBD seems to be preferred strategy over EST for endoscopic remove of common bile duct stones in patients who have coagulopathy. Although stone recurrence rate more than a year of EPBD is lower, but the overall stone clearance rate is lower and the risk of pancreatitis is higher than that of EST.
Dilatation ; Gallstones ; surgery ; Humans ; Postoperative Complications ; epidemiology ; Randomized Controlled Trials as Topic ; Sphincterotomy, Endoscopic ; Treatment Outcome
7.Analysis of posterior pedicle screw-only constructs in surgical treatment of adolescent idiopathic scoliosis with a minimum three-year follow-up.
Ming LI ; Ying-chuan ZHAO ; Xiao-dong ZHU ; Shi-sheng HE ; Chuan-feng WANG ; Chang-wei YANG
Chinese Journal of Surgery 2010;48(6):410-414
OBJECTIVETo evaluate clinical outcomes of a group of adolescent idiopathic scoliosis (AIS) patients undergoing posterior pedicle screw-only instrumentations.
METHODSBetween April 2002 and July 2006, 121 AIS patients (93 female and 28 male, average age at operation was 15.5 years which ranged from 10 to 20 years) received posterior pedicle screw-only instrumentation and fusion. All the patients were evaluated by the various-parameters measured in X-ray films before and after surgery, including Cobb angle on coronal plane, Cobb angle on sagittal plane, clavicle angle and shoulder height difference, lowest instrumented vertebrae (LIV) angulation, proximal junction kyphotic angle, the distances of central sacral vertical line (CSVL) to the LIV, to the apical vertebra and to the C(7) plumb line respectively. Complications were followed.
RESULTSAn average of (11.0 + or - 1.5) levels was fused. An average coronal correction of proximal thoracic curve was 41.8%, of thoracic curve was 70.8%, of thoracolumbar/lumbar curves was 74.0%. No significant change was found in sagittal alignment. Shoulder balance and apex vertebral to central sacral line were restored well. There were no pseudoarthroses and loss of correction during the follow-ups. One adding-on, 4 proximal thoracic decompensation and 15 proximal junction kyphosis were found during the follow-ups.
CONCLUSIONPosterior pedicle screw-only instrumentation and fusion has excellent radiographic and clinical results with minimal complications in the surgical treatment of AIS.
Adolescent ; Bone Screws ; Child ; Female ; Follow-Up Studies ; Humans ; Male ; Retrospective Studies ; Scoliosis ; surgery ; Spinal Fusion ; instrumentation ; methods ; Treatment Outcome ; Young Adult
8.Influence of Jiangu decoction on peroxisome proliferator-activated receptor (PPAR) in the femoral head of rabbits with steroid-induced femoral head necrosis.
Wen-Gang LIU ; Wei HE ; Xue-Meng XU ; Huai WU ; Ling-Feng WEI ; Chuan-Xi ZHAO
China Journal of Orthopaedics and Traumatology 2012;25(5):407-410
OBJECTIVEThe Jiangu decoction is used in the treatment of steroid-induced femoral head necrosis in clinical experiences, which has functions of tonifying kidney and activating blood, and invigorating spleen to remove phlegm. The decoction is mainly composed of Radix Polygoni Multiflori, Rhizoma alismatis Rhizoma Drynariae, haw, medlar, Radix Astragali, radix rehmanniae, angelica, Radix Codonopsis, radix salviae miltiorrhizae, Fructus Ligustri Lucidi, licorice, pharmaceutical composition. This study was designed to investigate the influence of Jiangu decoction on peroxisome proliferator-activated receptor gamma (PPARgamma) in the femoral head of rabbits with steroid-induced femoral head necrosis.
METHODSEighteen adult SPF healthy New Zealand rabbits were divided into 3 groups: control group, model group, Jiangu decoction group. The rabbits of Jiangu decoction group orally received Jiangu decoction suspension with a dose of 10 ml/kg each day and the drug content was 0.719 g/ml. The rabbits in control and model groups were given saline with a dose of 10 ml/kg. The methylprednisolone sodium succinate was injected intramuscularly into left leg with a dose of 40 mg/kg. Then the rabbits were fed continuously for 3 weeks. The glucocorticoid levels, PPARgamma and plasma glucocorticoid levels in the femoral head were measured before and after modeling.
RESULTSBefore model established, the plasma glucocorticoid levels had no significant difference among three groups (P=0.301). At 3 weeks after model established,the plasma glucocorticoid level of rabbits in model group increased compared to the control group (P=0.001); and the plasma glucocorticoid level of rabbits in Jiangu decoction group decreased compared with model group (P=0.001). The glucocorticoid level in the local femoral head of rabbits in model group increased compared to the control group (P=0.001); and the glucocorticoid level in the local femoral head of rabbits in Jiangu decoction group decreased compared with model group (P=0.001). The PPARgamma level in the local femoral head of rabbits in model group increased compared to the control group (P=0.018);and the PPARgamma level in the local femoral head of rabbits in Jiangu decoction group decreased compared with model group (P=0.033).
CONCLUSIONThe Jiangu decoction is effective to inhibit the femoral head adipogenic differentiation by decrease the PPAR content, so as to prevent and treat steroid-induced femoral head necrosis.
Animals ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Femur Head Necrosis ; chemically induced ; drug therapy ; pathology ; Glucocorticoids ; blood ; toxicity ; Male ; Medicine, Chinese Traditional ; PPAR gamma ; analysis ; Rabbits
9.Determination of positive bronchodilating response using impulse oscillation system in children.
Chuan-he LIU ; Shuo LI ; Xin SONG ; Chao CHEN ; Jing ZHAO ; Yu-zhi CHEN
Chinese Journal of Pediatrics 2005;43(11):838-842
OBJECTIVEImpulse oscillation system (IOS) as an approach to lung function determination is suitable for children, especially for preschool children in that it only requires the individual to be examined to breathing stably with tidal volume. However, until now there have been no uniformly agreed criteria for positive bronchial reversibility test in clinical practice. A screening method was applied to seek for answer when this question in the hope of providing objective evidence for clinical diagnosis of respiratory diseases, esp. asthma.
METHODSOne hundred and fifty-six children patients during asthma attack were recruited randomly for this study from January to December, 2004. These patients included 103 boys and 53 girls aged from 5 to 18 years with mean age of 8.84 +/- 2.58 years, who visited the doctors in the Clinical and Educational Centre for Asthma, Capital Institute of Pediatrics and were diagnosed as asthma. Maximal expiratory flow volume and IOS lung function were determined followed by bronchodilator reversibility test in all patients. The sensitivity and specificity of IOS parameters, total respiratory impedance (Zrs), respiratory resistance at 5 Hz (R5) and reactance at 5 Hz (X5) for diagnosing asthma at different improvement levels after inhalation of bronchodilator were calculated respectively using forced expiratory volume in first second (FEV(1)) and maximal mid-expiratory flow (MMEF) as gold standard. And the regression equation between parameters of lung function which were determined with the two different methods was analyzed.
RESULTSAfter inhalation of bronchodilator, the lung function of the children was improved significantly (P < 0.01), with the FEV(1), MMEF increase and Zrs, R5, and X5 decrease. There were significantly positive correlations (P < 0.01) between the improvement rate of parameters of lung functions determined with the two methods, and the highest correlation coefficients existed between X5 and FEV(1) and MMEF (respectively 0.676, 0.571), correlation coefficients between X5 and Zrs ranked second (0.519 and 0.505, respectively). When an increase of FEV(1) equal to or greater than 15%, or increase of MMEF equal to or greater than 30% was used as reference for positive bronchial reversibility, the sensitivity and specificity were relatively high for considering positive bronchial reversibility test with at least 20% decrease of Zrs, R5, and 30% decrease of X5 with IOS lung function. The sensitivity and specificity of Zrs, R5 and X5 calculated from FEV(1) were 0.62, 0.51; 0.49, 0.54; 0.70, and 0.54, respectively. The sensitivity and specificity of Zrs, R5 and X5 calculated from MMEF were 0.63, 0.54; 0.53, 0.60; 0.60, and 0.58, respectively. The regression equation showed that the decrease of Zrs, R5 and X5 corresponding to 15% increase of FEV(1) and 30% increase of MMEF were 21.7%, 21.3%; 19.9%, 19.5%; 30.1%, and 29.6%, respectively.
CONCLUSIONWhen lung function is determined with IOS in children, only when decrease of Zrs and R5 is equal to or more than 20% and decrease of X5 is 30% or more after inhalation of bronchodilator, can the bronchial reversibility test be considered as positive.
Adolescent ; Asthma ; diagnosis ; Bronchodilator Agents ; administration & dosage ; Child ; Child, Preschool ; Female ; Forced Expiratory Volume ; Humans ; Male ; Maximal Midexpiratory Flow Rate ; Regression Analysis ; Respiratory Function Tests ; methods ; Sensitivity and Specificity
10.Construction and characterization of invasion protein B gene deleted mutant asd host-vector balanced lethal system
Chunjie ZHANG ; Songbiao CHEN ; Xiangchao CHENG ; Chengshui LIAO ; Jing LI ; Lei HE ; Mingliang ZHANG ; Chuan YU ; Zuhua YU ; Yanyan JIA ; Zhanqin ZHAO
Chinese Journal of Immunology 2016;(2):210-213
Objective: To develop an oral live vaccine vector which stably carries exogenous genes.Methods:SL1344ΔsipBΔasd host-vector balanced lethal system was constructed by the method of recombinant suicide plasmid-mediated allelic exchange on the basis of attenuated Salmonella typhinurium SL1344ΔsipB.Then,the biological characteristics of SL1344ΔsipBΔasd was analyzed.Results:The results showed that the mutant was stabile with the Δasd gene in vitro;the serotype and growth rate of SL1344ΔsipBΔasd strain was almost same as the parent SL1344ΔsipB and SL1344 strain.And the mutant strains remain swim ming zones.Virulence test in mice showed that the virulence of SL1344ΔsipBΔasd which carried complementary plasmid pYA3493 by electro-transformation decreased by 1.4%compared with SL1344.Conclusion: These results showed that the SL1344ΔsipBΔasd mutant was successfully constructed.It is likely that this mutant should be used as a live vector to express foreign genes.