2.Differentiation of human umbilical cord mesenchymal stem cells into corneal epithelium like cells
Shu-yi, XU ; Guang-hui, HOU ; Jing, WU ; Jin-tang, XU
Chinese Journal of Experimental Ophthalmology 2012;(10):882-887
Background Corneal blindness caused by ocular surface disease is one of the main reasons for the global blinding corneal diseases.With the development and progress of tissue engineering technology,tissueengineered cornea offers a new approach to the treatment of ocular surface disease.Objective This study was to obscrve the growth and differentiation of human umbilical cord mesenchymal stem cclls (UC-MSCs) on thc corneal stroma of receipts and investigate the feasibility of human UC-MSCs differentiated into corneal epithelium-like cells and the reparation of injury cornea.Methods Human UC-MSCs were isolated from human umbilical cord using collagenase Ⅳ digestion and passaged in DMEM/F12 containing fetal bovine serum in vitro.The immunophenotype of cultured human UC-MSCs was evaluated by flow cytometry.The differentiated osteoblasts from the human UC-MSCs by directional induce was identified.Twenty-four New Zealand albino rabbits were randomly divided into 2 groups.The human UC-MSCs were cultured on porcine corneal matrix without corneal epithelium for 4 days and then transplanted onto the 12 left eyes of 12 New Zealand albino rabbits,and porcine corneal matrix without corneal epithelium was transplanted onto the left eyes of other 12 New Zealand albino rabbits as control group.The rabbits received keratoplasty were examined using in vivo confocal microscope through focusing(CMTF).The eyeballs were taken off after 2,4 and 8 weeks,the growth and differentiation,expression of cytokeratin 3 (CK3),CK12 and ATP-binding cassette superfamily G memben 2 (ABCG2)of human UC-MSCs were observed by histopathology and immunofluorescence staining.This use of the experimental animals complied with ARVO Statement.Results Digestive human UCMSCs formed round in shape and was large in size.The attached cells displayed long-fusiform shape like fibroblasts.The cultured human UC-MSCs phenotype was CD105+/CD29+/CD44+/CD34-/CD45-and could be induced toward osteoblast differentiation under the appropriate experimental conditions.Human UC-MSCs grew well on the porcine corneal matrix.The corneal grafts survived wcll without rejection till the experiment end in experimental eyes,but the rejection of corneal graft occurred in control eyes.Confocal microscope could observe corneal epithelium-like cells.The corneal epithelium cells showed the positive response for CK3 and CK12 and absent response for ABCG2.Conclusions Human UC-MSCs with porcine corneal matrix can survive,proliferate and differentiate into corneal epithelium-like cells after transplanting onto the corneal stroma of rabbits.This result suggests that human UC-MSCs is able to repair and reconstruct the injured corneal surfaces.
3.Observation of Dynamic Changes in Ultra-Micro-Structure of Pulmonary Arteries and Endogenous Hydrogen Sulfide in Rats with Left-Right Shunt
xiao-hui, LI ; jun-bao, DU ; xiu-ying, TANG ; hong-fang, JIN ; ya-guang, DING ; jian, LI ; chao-shu, TANG
Journal of Applied Clinical Pediatrics 2004;0(07):-
Objective To explore the relationship between dynamic changes in ultra-micro-structural of pulmonary arteries and endogenous hydrogen sulfide in rats with left-right shunt.Methods Rats in shunt group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of pulmonary artery structural remodeling. After 1 day, 3 days, 1 week, 4 weeks and 8 weeks of experiment, the ultra-micro-morphologic changes of pulmonary arteries of rats were observed under electronic microscope and H_2S concentration in serum was evaluated by modified sulfide electrode method.Results The changes of ultra-micro-structure of pulmonary arteries were progressively exacerbated, endothelial cells became swollen and large in size on 3 days, smooth muscular cells increased in size as well as the change of endothelial cells in 1 week, and they changed from contractile phenotype to synthetic phenotype in 4 weeks.Conclusions Shunt exhibited changes of ultra-micro-structure of pulmonary arteries are accompanied by the changes of endogenous H_2S. It is suggested that endogenous H_2S might play a protective role in changes of ultra-micro-structure of pulmonary artery.
4.Alterations of proadrenomedullin N-terminal 20-peptide in rats with pulmonary hypertension induced by high pulmonary blood flow.
Jian-Guang QI ; Xiao-Hui LI ; Ya-Guang DING ; Chao-Shu TANG ; Jun-Bao DU
Chinese Journal of Contemporary Pediatrics 2007;9(6):574-576
OBJECTIVEThe mechanism of high pulmonary blood flow-induced pulmonary hypertension remains unclear. The aim of this study was to explore the effect of proadrenomedullin N-terminal 20-peptide (PAMP) on pulmonary hypertension, through examining the alterations of pulmonary PAMP expression and plasma PAMP concentration in rats with pulmonary hypertension induced by high pulmonary blood flow.
METHODSSixteen male Sprague-Dawley rats were randomly divided into control (n=8) and shunt groups (n=8). Aortocaval shunting was produced in the shunt group. After 11 weeks of shunting, systolic pulmonary artery pressure (sPAP), diastolic pulmonary artery pressure (dPAP) and mean pulmonary artery pressure (mPAP) were evaluated by using a right cardiac catheterization procedure. The ultrastructural changes in intra-acinar pulmonary arteries were observed. The concentration of plasma PAMP was measured by radioimmunoassay. The expression of PAMP in pulmonary arteries was detected by immunohistochemical assay.
RESULTSsPAP, dPAP and mPAP were significantly increased in shunt rats compared with controls (P < 0.01). Ultrastructural changes, such as hyperplasia and swelling of endothelial cells, irregularity of internal elastic laminar, and hypertrophy and increased number of synthetic phenotype of smooth muscle cells, were found in intra-acinar pulmonary muscularized arteries in the shunt group. Plasma PAMP concentration (616 +/- 195 pg /mL vs 427 +/- 90 pg /mL) and PAMP expression in endothelial cells (0.62 +/- 0.09 vs 0.38 +/- 0.12) and in smooth muscle cells (0.24 +/- 0.07 vs 0.14 +/- 0.05) of pulmonary arteries increased significantly in the shut group compared with controls.
CONCLUSIONSThe up-regulation of pulmonary and plasm PAMP expression might be involved in the development of high pulmonary blood flow-induced pulmonary hypertension.
Adrenomedullin ; blood ; genetics ; Animals ; Hypertension, Pulmonary ; blood ; etiology ; pathology ; Male ; Pulmonary Artery ; ultrastructure ; Pulmonary Circulation ; Rats ; Rats, Sprague-Dawley
5.Electrocardiographic characteristics and radiofrequency catheter ablation results of ventricular tachycardia or premature ventricular contractions originating in the vicinity of atrioventricular annulus.
Jin-lin ZHANG ; Xi SU ; Hong-wei HAN ; Zhen LI ; Cheng TANG ; Ping JIANG ; Guang-hui CHENG
Chinese Journal of Cardiology 2008;36(12):1083-1086
OBJECTIVETo evaluate the electrocardiographic (ECG) characteristics and results of radiofrequency catheter ablation (RFCA) of idiopathic ventricular tachycardia or premature ventricular contractions (VT/PVCs) originating in the vicinity of atrioventricular annulus.
METHODSNineteen patients with idiopathic VT/PVCs underwent conventional electrophysiological study and RFCA were included in this analysis. The 12 leads (ECG) characteristics were also analyzed.
RESULTSThe VT/PVCs were originated in the vicinity of mitral annulus in 10 cases, including anterolateral (n = 5), posterolateral (n = 3) and posteroseptal (n = 2). The VT/PVCs were originated in the vicinity of tricuspid annulus in the rest 9 cases, including the free wall (n = 5) and the septal portion (n = 4). The 12-lead ECG patterns of VT/PVCs originating in the vicinity of atrioventricular annulus were helpful for determine the site of RFCA. The VT/PVCs were successfully eliminated by RFCA in all cases.
CONCLUSIONThe exact origin of VT/PVCs originating in the vicinity of atrioventricular annulus could be determined by 12-leads ECG analysis and can be successfully and safely cured by RFCA.
Adult ; Aged ; Catheter Ablation ; methods ; Electrocardiography ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tachycardia, Ventricular ; etiology ; physiopathology ; therapy ; Treatment Outcome ; Tricuspid Valve ; physiopathology ; Ventricular Premature Complexes ; etiology ; physiopathology ; therapy
6.Hepatic angiomyolipoma:correlation of contrast-enhanced ultrasound, contrast-enhanced CT and pathologic ifndings
Yi, ZHANG ; Rui, LI ; Xiao-hang, ZHANG ; Wei, CHEN ; Chuan-ming, LI ; Guang-jie, DUAN ; Yan-li, GUO ; Chun-lin, TANG ; Zhao-hui, CHEN
Chinese Journal of Medical Ultrasound (Electronic Edition) 2013;(11):929-934
Objective To compare the features of contrast enhanced ultrasound (CEUS) and contrast-enhanced computed tomography (CECT) of hepatic angiomyolipoma (HAML), and to explore their relationship with pathologic findings. Methods Thirty patients with 31 resected or punctured and pathologically proved hepatic angiomyolipomas in Southwest Hospital of Third Military Medical University from January 2006 to December 2012 were selected in this retrospective study. CEUS and CECT features were evaluated and analyzed with pathology findings in 30 patients with HAML preoperatively. The proportion of typical performance by CEUS compared with CECT in this group was analyzed with Fisher exact propability. Results Seventeen lesions were inhomogeneous hyperechoic under conventional ultrasound observation. Twenty lesions demonstrated typical imaging characteristics by CEUS, eleven lesions showed atypia CEUS imaging characteristics. There were seventeen lesions on CT indicates the presence of fat. Seven lesions demonstrated typical imaging characteristics by CECT, twenty-four lesions presented atypical CECT imaging characteristics. The proportion of showing typical imaging characteristics by CEUS was higher than by CECT (64.5%vs 22.6%, P=0.002). Among the eleven mixed type HAML lesions, seven lesions showed typical CEUS imaging characteristics and two lesions demonstrated typical CECT imaging characteristics. In the ten myomatous type HAML lesions, six lesions displayed typical CEUS imaging characteristics and two lesions revealed typical CECT imaging characteristics. Among the eight lipomatous type HAML lesions, six lesions showed typical CEUS imaging characteristics and three lesions displayed typical CECT imaging characteristics. Conclusions Conventional ultrasound combining with CEUS can demonstrate the echoic and blood perfusion characteristics of HAML in most cases. The features of CEUS and CECT were varied in different histological types.
7.Logistic regression analysis of the risk factors in progressive hemorrhagic injury after severe head injury
Guang FENG ; Ji-Xin DUAN ; Zhi-Jun ZHONG ; Han-Chang YU ; Ling HAN ; Hui TANG
Chinese Journal of Neuromedicine 2010;09(11):1150-1153
Objective To study the risk factors related with progressive hemorrhagic injury (PHI)after severe head injury for a view to early diagnosis and treatment for this disease and providing a basis for effective prevention. Methods In a retrospective study of 262 patients with severe brain injury in considering the clinical data of the PHI, the occurrence is variable, and age, gender, bleeding site, type of bleeding, dilated pupils starus, level of systolic blood pressure on admission, time of CT for the first time,GOS scores, injured mechanism, interval between first and second time CT, application of high-dose mannitol, platelet (PLT) count, prothrombin time (PT), activated partial prothrombin time (APTT) were considered as independent variables. Results The incidence rate of having PHI was 47.7% (125/262);single-factor analysis revealed that, as compared with those in patients with non-PHI, 7 factors in patients with PHI were significantly different, namely, age, type of hemorrhage, interval between injury and first-time CT, GCS scores, PLT count, PT and APTT. Multivariate logistic regression analysis of the results showed that interval between injury and first-time CT, GCS scores, PLT count were the risk factors of having PHI, and their OR values were 3.5448, 3.2975 and 2.2361, respectively. Conclusion For patients with severe brain injury, the sooner the first time CT examination is performed, the lower the GCS scores are and the lower the PLT count is, the higher risk of having PHI is. Thus, dynamic CT formal review is suggested to improve the early diagnosis and treatment of PHI.
8.Matrine inhibits the proliferation of neuroblastoma LA-N-5 cell and MYCN mRNA expression.
Chen FENG ; Suo-Qin TANG ; Jian-Wen WANG ; Hui LONG ; Guang YANG
Chinese Journal of Contemporary Pediatrics 2008;10(2):225-227
OBJECTIVENeuroblastoma is the most common malignant solid tumor in children under 4 years. Amplification of MYCN oncogene is associated with advanced-stage disease, rapid tumor progression, resistance to treatment, and poor outcome. Matirne has the anti-tumor activity. This study was designed to investigate the effects of matrine on LA-N-5 cell line proliferation and MYCN gene mRNA expression.
METHODSNeuroblastoma LA-N-5 cells were treated by 0.25, 0.50, 0.75 or 1.00 mg/mL matrine. MTT was used to measure the levels of the proliferation of LA-N-5 cells cultured with different concentrations of matrine. MYCN gene mRNA expression in LA-N-5 cells was measured using real time RT-PCR with SYBR GREEN I fluorescence.
RESULTSThe proliferation of LA-N-5 cells was obviously inhibited by matrine in a dose- and time- dependent manner. Matrine of 1.00 mg/mL treatment for 72 hrs produced a best effect, with an inhibitory rate of LA-N-5 cell proliferation of 36.3% and an inhibitory rate of MYCN gene mRNA expression of 44.6%.
CONCLUSIONSMatrine may inhibit the growth of neuroblastoma cells and down-regulate MYCN mRNA expression. It may be promising as a new drug for treatment of neuroblastoma.
Alkaloids ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; N-Myc Proto-Oncogene Protein ; Neuroblastoma ; drug therapy ; metabolism ; pathology ; Nuclear Proteins ; genetics ; Oncogene Proteins ; genetics ; Quinolizines ; pharmacology ; RNA, Messenger ; analysis
9.Clinical epidemiologic features of burns in 2894 hospitalized children from Jinzhou of Northeast China.
Jie XIAO ; Hai-Jun WANG ; Guang-Hui HAO ; Feng TANG
Chinese Journal of Contemporary Pediatrics 2010;12(4):298-300
Burns
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epidemiology
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Child
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Child, Preschool
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China
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epidemiology
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Female
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Hospitalization
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Humans
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Male
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Seasons
10.Long-term observation of large weight-bearing bone defect in goats repaired with tissue engineering technique.
Bin CHEN ; Guo-xian PEI ; Ke WANG ; Guang-hui TANG
Journal of Southern Medical University 2006;26(6):770-773
OBJECTIVETo observe the long-term effect of tissue engineering-based repair of large weight-bearing bone defect in goats, and the final outcome of the scaffold material coral hydroxyapatite (CHAP) in vivo.
METHODSFifteen Chinese goats were subjected to operations to induce a 2-cm left tibial diaphyseal defect, which was filled subsequently with CHAP and bone marrow stromal stem cells (BMSCs). The repaired defects were evaluated by ECT, X-ray and histology in the early stage and at 6, 12, 18, and 24 months postoperatively.
RESULTSECT showed good bone regeneration and revascularization within 2 months postoperatively. X-ray and histology displayed eccentric and gradual bone regeneration in the early stage, and the tissue-engineered bone graft was firmly healed with the goat tibia. X-ray and histological examination at 6, 12, 18, 24 months postoperatively revealed moulding of the new bones and medullary cavity recanalization, and the structure of CHAP disappeared and gradually integrated into the new bones.
CONCLUSIONTissue-engineered bone is capable of total repair of large bone defect in goats by forming normal functional new bones. CHAP can be eventually degraded completely and become the component of the newly generated bones.
Animals ; Bone Marrow Transplantation ; Bone Regeneration ; Bone Substitutes ; Cells, Cultured ; Goats ; Prostheses and Implants ; Tibial Fractures ; physiopathology ; surgery ; Tissue Engineering ; methods