To develop an ideal surgical procedure for neobladder reconstruction in experimental porcine models. Six experimental female pigs weighting 28-33 kg underwent transplantation of autologous peritoneum for bladder reconstruction under general anesthesia.The flaps were used to reconstruct the orthotopic neobladder by suturing with the edges of the triangle and neck of the remnant bladder.The ureteral catheters were removed on the 5 postoperative day and the balloon catheter was removed on the 7 postoperative day.Voiding behaviour was monitored.The animals were euthanized at week 12 for routine pathology,immunohistochemistry,and electron microscopy. All the pigs survived after the surgery,and no postoperative complication such as peritonitis,intestinal obstruction,or urinary fistula was observed.All the peritoneum-ileum composite free valves survived after transplantation.Voiding behaviour was normal after catheter removal,and the urine was clear.At autopsy,reconstructed bladders were healthy.Pathological examination showed the neobladder had been covered by continuous urothelium while the peritoneum disappeared and showed no ileal mucosa regrowth and residual.Scanning electron microscope showed the transitional cells of neobladder were complete and orderly,and the urothelium around suture border was continuous and showed no malposition. Reconstruction of bladder by autologous peritoneum and ileal seromuscular flaps is an ideal approach in the experimental pigs as it can prevent regrowth of ileal epithelial cells and avoid the complications of conventional enterocystoplasty.Its clinical application deserves further investigations.
Animals ; Cystectomy ; Female ; Ileum ; Peritoneum ; Postoperative Complications ; Surgical Flaps ; Swine ; Urinary Bladder Neoplasms

BACKGROUNDTo overcome the drawbacks of permanent stents, biodegradable stents have been studied in recent years. The bioabsorbable polymer vascular scaffold (BVS) was the first bioabsorbable stent to undergo clinical trials, demonstrating safety and feasibility in the ABSORB studies. Iron can potentially serve as the biomaterial for biodegradable stents. This study aimed to assess the short-term safety and efficacy of a biodegradable iron stent in mini-swine coronary arteries.

METHODSEight iron stents and eight cobalt chromium alloy (VISION) control stents were randomly implanted into the LAD and RCA of eight healthy mini-swine, respectively. Two stents of the same metal base were implanted into one animal. At 28 days the animals were sacrificed after coronary angiography, and histopathological examinations were performed.

RESULTSHistomorphometric measurements showed that mean neointimal thickness ((0.46 ± 0.17) mm vs. (0.45 ± 0.18) mm, P = 0.878), neointimal area ((2.55 ± 0.91) mm(2) vs. (3.04 ± 1.15) mm(2), P = 0.360) and percentage of area stenosis ((44.50 ± 11.40)% vs. (46.00 ± 17.95)%, P = 0.845) were not significantly different between the iron stents and VISION stents. There was no inflammation, thrombosis or necrosis in either group. The scanning electron microscopy (SEM) intimal injury scores (0.75 ± 1.04 vs. 0.88 ± 0.99, P = 0.809) and number of proliferating cell nuclear antigen (PCNA) positive staining cells were not significantly different between the two groups. The percentage of neointimal coverage by SEM examination was numerically higher in iron stents than in VISION stents ((84.38 ± 14.50)% vs. (65.00 ± 22.04)%, P = 0.057), but the difference was not statistically significant. Iron staining in the tissue surrounding the iron stents at 28 days was positive and the vascular wall adjacent to the iron stent had a brownish tinge, consistent with iron degradation. No abnormal histopathological changes were detected in coronary arteries or major organs.

CONCLUSIONSThe biodegradable iron stent has good biocompatibility and short-term safety and efficacy in the miniswine coronary artery. Corrosion of iron stents is observed at four weeks and no signs of organ toxicity related to iron degradation were noted.

Absorbable Implants ; adverse effects ; Animals ; Coronary Vessels ; surgery ; Iron ; chemistry ; Microscopy, Electron, Scanning ; Stents ; adverse effects ; Swine

Objective To investigate the polymorphism of human cytomegalovirus（HCMV）UL146 gene in clinical strains,and to evaluate its clinical diagnostic and therapeutic value of gene.Methods The UL146 gene of clinical strains was examined by quantitative polymerase chain reaction（Q-PCR）or general polymerase chain reaction（PCR）.Positive samples of PCR amplification were sequenced and analyzed.Results High variability of UL146 gene was found among 28 HCMV strains.According to phylogenetic analysis,all sequences of UL146 in clinical strains could be divided into three types and four subtypes.Chemokine ELRCXC region was highly conserved in all sequences.Conclusion HCMV-UL146 genes showed a high degree of polymorphism,and its encoded chemokine ELRCXC region was highly con-served.The relationship between HCMV-UL146 gene′s polymorphism and different clinical symptoms of HCMV infection was unclear.

OBJECTIVETo simulate and assess the clinical effect of intracoronary infusion of bone marrow mononuclear cells or peripheral endothelial progenitor cells on myocardial reperfusion injury in mini-swine model.

METHODSTwenty-three mini-swine with myocardial reperfusion injury were used as designed in the study protocol. About (3.54 +/- 0.90) x 10(8) bone marrow mononuclear cells (MNC group, n = 9) or (1.16 +/- 1.07) x 10(7) endothelial progenitor cells (EPC group, n = 7) was infused into the affected coronary segment of the swine. The other mini-swine were infused with phosphate buffered saline as control (n = 7). Echocardiography and hemodynamic studies were performed before and 4 weeks after cell infusion. Myocardium infarction size was calculated. Stem cell differentiation was analyzed under a transmission electromicroscope.

RESULTSLeft ventricular ejection fraction dropped by 0% in EPC group, 2% in MNC group, and 10% in the control group 4 weeks after cell infusion, respectively (P < 0.05). The systolic parameters increased in MNC and EPC groups but decreased in the control group. However, the diastolic parameters demonstrated no significant change in the three groups (P > 0.05). EPC decreased total infarction size more than MNC did (1.60 +/- 0.26 cm2 vs. 3.71 +/- 1.38 cm2, P < 0.05). Undermature endothelial cells and myocytes were found under transmission electromicroscope.

CONCLUSIONSTransplantation of either MNC or EPC may be beneficial to cardiac systolic function, but might not has obvious effect on diastolic function. Intracoronary infusion of EPC might be better than MNC in controlling infarction size. Both MNC and EPC may stimulate angiogenesis, inhibit fibrogenesis, and differentiate into myocardial cells.

Animals ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; Cell Differentiation ; Endothelial Cells ; cytology ; Myocardial Reperfusion Injury ; pathology ; therapy ; Myocardium ; pathology ; Stem Cells ; cytology ; Swine ; Swine, Miniature

OBJECTIVETo study the relationship between inflammation and neointimal proliferation after coronary stent implantation in porcine model.

METHODSTwenty normal minipigs were randomly divided into group A (implanted with 316L bare metal stents), group B (implanted with 605L bare metal stents), group C (implanted with PLGA coating 605L stents), and group D (implanted with rapamycin-loaded PLGA coating 605L stents). Each minipig was implanted with two same stents in left anterior descending artery and right coronary artery. Four weeks later, the animals were sacrificed and histomorphometric measurements on the stent-segment coronary arteries were made to calculate the correlation between inflammation area and neointimal area.

RESULTSGroup D had the smallest neointimal area [(0.64 +/- 0.38) mm2, P < 0. 001] and inflammation area (median 0.00 mm2, P = 0.009) among all the groups, while there were no statistical differences among group A, B, and C in neointimal area [(2.09 +/- 0.90), (2.11 +/- 1.07), and (1.42 +/- 0.35) mm2 respectively] and in inflammation area (0.22 , 0.21, and 0.09 mm2, respectively). Bivariate correlation analysis showed that the inflammation area was positively correlated with the neointimal area (P < 0.001, correlation coefficient = 0.719). When stent type, mean injury score, and EEL area were adjusted, partial correlations analysis showed that the inflammation area was still positively correlated with the neointimal area (P = 0.01, correlation coefficient = 0.498).

CONCLUSIONInflammation promotes the neointimal proliferation after coronary stent implantation. Sirolimus-eluting stent may reduce the inflammatory response.

Animals ; Coronary Vessels ; pathology ; Drug-Eluting Stents ; adverse effects ; Inflammation ; pathology ; Neointima ; pathology ; Stents ; adverse effects ; Swine ; Swine, Miniature ; Tunica Intima ; pathology

OBJECTIVETo investigate the therapeutic effectiveness of intracoronary implantation of autologous bone marrow mononuclear cells (BM-MNC) in miniswine model of reperfused myocardial infarction.

METHODSSixteen miniswine myocardial ischemic reperfusion injury models made by ligation of the distal one third segment of left anterior descending artery for 90 minutes were randomized into 2 groups. In BM-MNC group (n = 9), (3.54 +/- 0.90) X 10(8) BM-MNC were intracoronary injected, and in the control group (n = 7), phosphate buffered saline was injected by the same way. Echocardiographic and hemodynamic results, vessel density, and myocardial infarction size were evaluated and compared before and 4 weeks after cell transplantation.

RESULTSIn BM-MNC group, there were no differences between before and 4 weeks after transplantation in aspects of left ventricular ejection fraction (LVEF), interventricular septal thickness, left ventricular lateral and anterior septal wall thickness, cardiac output, or +dp/dtmax. In control group, LVEF, interventricular septal thickness, left ventricular lateral and anterior septal wall thickness, cardiac output, and +dp/dtmax decreased significantly 4 weeks after transplantation (P < 0.05). Left ventricular end-diastolic pressure and -dp/dtmax, did not change significantly before and after cell transplantation in both groups. Capillary density in BM-MNC group was greater than that in control group [(13.39 +/- 6.96)/high power field vs. (3.50 +/- 1.90)/high power field, P < 0.05]. Infarction area assessed by tetrazolium red staining and the infarction percentage decreased in BM-MNC group compared with those in control group (P < 0.05).

CONCLUSIONSTransplantation of BM-MNC into myocardium with ischemic reperfusion injury increases capillary density and decreases infarction area. It has significantly beneficial effect on cardiac systolic function rather than on diastolic function.

Animals ; Bone Marrow Cells ; cytology ; physiology ; Bone Marrow Transplantation ; Capillaries ; physiology ; Echocardiography ; Heart ; anatomy & histology ; physiology ; physiopathology ; Hemodynamics ; Myocardial Ischemia ; Random Allocation ; Swine ; Systole ; physiology ; Transplantation, Autologous ; physiology

OBJECTIVETo investigate the therapeutic effectiveness of intracoronary transplantation of autologous bone marrow mononuclear cells (BM-MNC) on myocardial ischemia reperfusion injury in mini-swine model.

METHODSMyocardial ischemia reperfusion injury model was established by ligating in 16 mini-swines, which were further randomized into two groups: (3.54 +/- 0.90) x 10(8) BM-MNC was intracoronarily transplanted in BM-MNC group (n = 9), and phosphate buffer saline was intracoronarily applied in the control group (n = 7). Ultrasonic cardiograhpy, hemodynamics, neovascular density, and myocardium infarction size were evaluated before and 4 weeks after transplantation.

RESULTSIn BM-MNC group, left ventricular ejection fraction (LVEF), intra-ventricular septa, lateral wall and anterior wall, cardiac output (CO) and + dp/dt(max) had no significant differences before and 4 weeks after transplantation (P > 0.05). In the control group, LVEF, intraventricular septa, lateral wall and anterior wall, CO, and + dp/dt(max) significantly decreased 4 weeks after transplantation (P < 0.05). Left ventricular end-diastolic pressure and- dp/dt(max) had no significant differences before and after cell transplantation. Capillary density was significantly larger in the BM-MNC group than in the control group [(13.39 +/- 6.96) /HP vs. (3.50 +/- 1.90) /HP]. The percentage and size of myocardial infarction was significantly lower in the BM-MNC group than in the control group.

CONCLUSIONTransplantation of BM-MNC into the myocardial ischemic reperfusion-injury area can increase capillary density and decrease infarction area, and thus remarkably improve cardiac systolic function.

Animals ; Bone Marrow Transplantation ; Coronary Vessels ; Myocardial Reperfusion Injury ; pathology ; physiopathology ; therapy ; Myocardium ; pathology ; Random Allocation ; Swine ; Swine, Miniature

OBJECTIVEThe development of pulmonary vascular bed is strongly flow-dependent. Abnormal pulmonary blood flow leads to pulmonary pathological changes. This study aimed to observe the pathological changes of small pulmonary arteries and alveoli in complex congenital heart defect with diminished pulmonary blood flow but without aortopulmonary collateral artery (APCA) and patent ductus arteriosus (PDA) in infants and young children.

METHODSAutopsy pulmonary specimens obtained from 5 children who died of non-cardiovascular diseases were used as the control group (age: 4-18 months). Fifty-six children (age: 4-36 months) with complex congenital heart defect with diminished pulmonary blood flow but without APCA and PDA served as the study group, including 34 cases of tetralogy of Fallot, 7 cases of double outlet right ventricle with pulmonary stenosis, 9 cases of single ventricle with pulmonary stenosis, 4 cases of tricuspid atresia with pulmonary stenosis and 2 cases of complete atrioventricular septal defect with pulmonary stenosis. Pulmonary specimen sections were stained by hematoxylin-eosin and Weigert-Van Gieson. Percentage of media thickness (MT%), percentage of media section area (MS%), number of small arterial per square centimeter (APSC), mean alveolar number (MAN), mean linear intercept (MLI), proportion of parenchyma area in total area (PPA%) and alveolar to small arterial ratio per unit area (AAR) were measured by morphologic quantitative analysis.

RESULTSMT% (10.93+/-2.87% vs 15.08+/-2.51%), MS% (18.97+/-5.56% vs 25.04+/-3.87%) and APSC (202.43+/-67.45 vs 441.69+/-65.29) decreased significantly in the study group compared with the control group (P<0.01). The internal diameter of small pulmonary artery (80.26+/-21.57 microm vs 58.53+/-10.29 microm; P<0.05), AAR (46.59+/-14.43 vs 34.46+/-4.98; P<0.01) and MLI (144.98+/-44.87 microm vs 108.39+/-20.76 microm; P<0.05) increased significantly compared with the control group.

CONCLUSIONSThe media of small pulmonary arteries becomes thinner, the lumen of small pulmonary arteries becomes larger, and the number of small arterial per square centimeter and the mean alveolar number are reduced in infants and young children with complex congenital heart defect with diminished pulmonary blood flow but without APCA and PDA.

Aorta ; abnormalities ; Child, Preschool ; Collateral Circulation ; Ductus Arteriosus, Patent ; pathology ; Female ; Heart Defects, Congenital ; pathology ; Humans ; Infant ; Lung ; pathology ; Male ; Pulmonary Artery ; abnormalities ; Pulmonary Circulation

OBJECTIVETo investigate the relationship between pulmonary pathological features and clinical physiology of congenital heart defects (CHD) with decreased pulmonary artery blood flow.

METHODSBetween July 2001 and May 2006, 18 patients with CHD with decreased pulmonary artery blood flow undergoing palliative or definitive repair and having lung biopsy intraoperatively were enrolled in this study. The patients' age was 0.4 - 8.0 years, and body weight was 6.0 - 20.0 kg. The method of semi-quantitative morphometric technique and an image analyzer were applied to measure the following indices of pulmonary microvessels: the percentage of media thickness (MT%), the percentage of media section area (MS%) and numbers of microvessels per square centimeter (VPSC). The diameters of left pulmonary artery (LPA) and right pulmonary artery (RPA) were measured with two-dimensional echocardiography. The percutaneous oxygen saturation (SpO(2)), hemoglobin concentration (HB) and hematocrit value (HCT) were examined and recorded preoperatively.

RESULTSThere was a significant negative correlation between SpO(2) and HCT or Hb (R(2) = 0.4914, P = 0.001 and R(2) = 0.5505, P < 0.001), the variation trend of these three variables was linked. There was a negative correlation between SpO(2) and the body weight (R(2) = 0.2208, P = 0.049), which is in accordance with clinical features of aggravated process of cyanosis and hypoxia. The morphological observation of lung biopsy specimens indicated that most of peripheral pulmonary arteries were distended, irregular and their walls were uneven, and "lake" type of pulmonary AV malformations were observed. There was a positive correlation between VPSC and the body weight or BSA (R(2) = 0.5472, P < 0.001 and R(2) = 0.5233, P = 0.001). There was a significant correlation between VPSC and LPA or RPA (R(2) = 0.4312, P = 0.003 and R(2) = 0.2463, P = 0.036). It was shown that the diameter of central pulmonary arteries could be a reflection of peripheral pulmonary artery growth. The diameter of LPA also correlated with the diameter of RPA (R(2) = 0.286, P = 0.022).

CONCLUSIONSFor patients with congenital heart defects with decreased pulmonary blood flow, the pulmonary pathological changes are the bases of their clinical physiologic features. It is suggested that they should be treated in their earlier stage of life.

Child ; Child, Preschool ; Female ; Heart Defects, Congenital ; pathology ; physiopathology ; Humans ; Infant ; Lung ; blood supply ; Male ; Pulmonary Artery ; abnormalities ; pathology ; physiopathology

OBJECTIVETo compare the effect of different antegrade pulmonary blood flow on the further development of pulmonary artery after Glenn procedure in cyanotic congenital heart defects (CHD) patients.

METHODSBetween October 2000 and December 2006, 132 CHD patients with decreased pulmonary artery blood flow underwent bidirectional Glenn shunt, among them 18 patients received intraoperative lung biopsy. Patients were divided into two groups according to their different sources of antegrade pulmonary blood flow: antegrade arterial blood flow group (n = 33) and antegrade venous blood flow group (n = 99). The percutaneous oxygen saturation (SpO2), hemoglobin (Hb) concentration, and hemotocrit (Hct) value were examined and recorded before and after operation. The diameters of left pulmonary artery (LPA) and right pulmonary artery (RPA) were measured with two-dimensional echocardiography and the results were used to calculate the pulmonary artery index (PAI). The method of half-quantitative morphometric technique and an image analyzer were used to measure the following indicators of pulmonary microvessels: the percentage of media thickness (MT%), the percentage of media section area (MS%), vascular numbers of per square centimeter (VPSC), and mean alveolar number (MAN).

RESULTSBefore the operation, obvious cyanosis was found in both groups, while SpO2, Hct, and Hb were not significantly different (P > 0.05). LPA, RPA, and PAI were not significantly different between two groups (P > 0.05). The MT% and MS% in antegrade venous blood flow group were significantly less than those in antegrade arterial blood flow group (P < 0.05), but VPSC and MAN were not significantly different (P > 0.05). After Glenn procedure, hypoxia and cyanosis were remarkably improved in both two groups. There was a significantly negative correlation between SpO2 and Hct (r = -0.49, P < 0.01) or Hb (r = -0.196, P < 0.01 ). The PAI increased by 22% in antegrade arterial blood flow group and 44% in antegrade venous blood flow group (P < 0.05). The diameters of LPA and RPA in antegrade venous blood flow group were significantly larger than those in antegrade arterial blood flow group (P < 0.05) and the growth of RPA in antegrade arterial blood flow group was not significant.

CONCLUSIONA better pulmonary artery growth occurs in the patients of pulmonary stenosis after Glenn shunt than in those of pulmonary atresia, and it contributes to an earlier completion of Fontan procedure.

Blood Flow Velocity ; Cardiac Surgical Procedures ; Child ; Child, Preschool ; Female ; Heart Defects, Congenital ; physiopathology ; surgery ; Humans ; Infant ; Male ; Pulmonary Artery ; growth & development ; physiopathology ; surgery ; Pulmonary Veins ; physiopathology ; Treatment Outcome