Objective To observe the morphology and growing status of mesenchymal stem cells(MSCs) of human bone marrow in vitro, in order to confirm that MSCs of human bone marrow are ideal seed cells and provide basic theory for further MSCs research. Methods The methods of density gradient centrifugation with lymphocyte separation medium for human and adherent filtration were used to isolate and purify MSCs of human bone marrow. We observed the cellular growth status and morphology of the primary MSCs and the surface antigens of second passage MSCs were tested. Results The primary culture cells fused into monolayer after 14-16 d. The passage cells kept the same morphological characteristics of primary culture cells. Ultrastructure of the second passage MSCs showed that the shape of nuclei was irregular, there were multiple nucleoli in some of the nuclei, and morphological differentiation of intracytoplasm organelles was immature. The growth curve of the first, fifth and tenth passage cells showed a logarithmic growth at day 3, a peak growth at day 5, and no clones occurred after tenth passage. Cloning efficiency of first passage, fifth passage and tenth passage was respectively 25.83%±2.93%, 14.67%±1.63% and 4.67%±0.52%. Test of MSCs phenotypic characteristics showed a high homogeneity among the cells and surface antigen profiles were positive for CD29, CD44 and negative for CD34, CD45. Conclusion The methods of density gradient centrifugation with lymphocyte separation medium for human and adherent filtration are simple, economic and efficient to isolate and purify MSCs from human bone marrow. With a high proliferating ability in vitro, MSCs from human bone marrow are ideal seed cells for tissue engineers.
Objective To report the surgical treatment for double outlet of ventricle with atrioventricular discordance. Methods 11 patients aged from 3 to 25 years underwent surgical treatment for double outlet of ventricle with atrioventricular discordance. 10 of them were double outlet right ventricle and the other one was double outlet left ventricle. The surgical treatment included biventricular repair (n=9) and single ventricular repair (n=2). The biventricular repair was performed by intraventricular patch repair and extracardiac valved conduit or homograft valved conduit. The single ventricular repair included bidirected Glenn operation and total cavopulmonary connect. Results There were two early deaths in biventricular repair and no death in single ventricular repair. The cause of death was severe low cardiac output syndrome. Four survivors in biventricular repair were followed up from 1 to 11.5 years, three of them were in NYHA class Ⅰ or Ⅱ and one in NYHA class Ⅲ. Conclusions Double outlet of ventricle with atrioventricular discordance can be treated by biventricular repair or single ventricular repair according to the development of ventricle and pulmonary arteries. Closure of ventricular septal defect , no obstruction from ventricle to great artery and no harm of conduction system are the keys of conventional management to achieve good results.
ObjectiveTo study the changes of levels of α subunits of stimulatory (Gsα) and inhibitory guanine nucleotide binding protein (Giα) in newborn guinea pig (0 2 days old) myocardium undergoing global ischemic reperfusion, and influences on the changes by St.Thomas Ⅱ and cold blood cardioplegic solution.MethodsThirty newborn guinea pigs were randomly assigned to three groups. GroupⅠ ( n = 10): the newborn hearts suffered by hypothermic global ischemia; group Ⅱ( n =10): the newborn hearts arrested by St. Thomas Ⅱ , and group Ⅲ ( n = 10): the newborn hearts arrested by cold blood cardioplegic solution. Levels of Gsα and Giα were investigated with Western blot analysis.ResultsNo differences of levels of Gsα and Giα were found in three groups before ischemia ( P gt;0.05). The level of Gsα after ischemia was significantly decreased than before ischemia in groupⅠand group Ⅱ ( P lt; 0 01), whereas no pronounced changes in group Ⅲ ( P gt;0.05) were noted after ischemia. The level of Gsα in group Ⅲ was not significantly changed after reperfusion compared with before ischemia( P gt;0 05), and it was much higher than those in groupⅠand group Ⅱ ( P lt; 0 01). Level of Giα was found not markedly changed in group Ⅲ after reperfusion compared with that before ischemia, but was notable higher in groupⅠand group Ⅱ( P lt;0.01). ConclusionsSignificant decrease of level of Gsα, whereas marked increase of level of Giα are found in myocardium of newborn guinea pig undergoing hypothermic (20℃) ischemic reperfusion. No impact of St. Thomas Ⅱ on these changes is verified, but recovery to the level of Gsα and Giα before ischemia is achieved by cold blood cardioplegic solution after ischemia and reperfusion. Unbalance between Gsα and Giα is the one of the mechanisms of ischemic reperfusion injury for immature myocardium.
Objective To summarize the experience of surgical correction of complete atrioventricular septal defect associated with tetralogy of Fallot(CAVSD-TOF). Methods Twelve patients aged 6-16(11.1±2.8) years underwent correction of CAVSD-TOF. The atrioventricular septal defect was closed through a right atriotomy and longitudinal right ventriculotomy in each case. The three-patch technique was used for the first 7 cases and two-patch technique for the later 5 cases. The commissure between the superior and inferior bridging leaflets of the left portion of the common atrioventricular valve was closed in each patient. Right ventricular outflow tract obstruction was relieved by a transannular patch. Results There were 4 deaths in the early postoperative period, 3 deaths in the first 7 cases compared to 1 death in the later 5 cases. The causes of death included severe low cardiac output syndrome(3 cases) and perfusion pulmonary edema(1 case). Six survivors were followed up from 3 months to 13.5 years. Heart function (NYHA) was class I or Ⅱ in all cases. Conclusion CAVSD-TOF can be corrected by using the two-patch technique and closure of atrioventricular septal defect through a combined approach through right atriotomy and right ventriculotomy. Routine closure of the commissure of the left portion of the atrioventricular valve achieves a low incidence of regurgitation.
Objective To summarize the surgical experience of supracardiac total anomalous pulmonary venous connection(S-TAPVC) and study the surgical technique and outcomes for S -TAPVC. Methods Eightysix patients with S-TAPVC underwent the surgical repair from May 1985 to December 2007. There were 49 males and 37 females. The patients aged from 7 months to 35 years (mean 9.6 years) and weighed from 4.9 kg to 68.0 kg (mean 23.8 kg). The patients were divided into three groups by the approach for the anastomosis. There were 20 patients in groupⅠthrough the right atrium incision, 49 patients in group Ⅱ through the right and left atrium incisions and 17 patients in group Ⅲ through the top of the left atrium incision. The interrupt continuous anastomosis between the common pulmonary venous and the left atrium was used in all patients. The enlarged atrial septal defect(ASD) was repaired with autopericardium. The vertical vein was ligated if the postoperative left atrial pressure was less than 15 mm Hg. But the vertical vein was opened or just partialy ligated if the postoperative left atrial pressure was more than 15 mm Hg. Results There was no early operative death. The postoperative left atrial pressure in three groups were 9.3±3.2 mm Hg, 9.9±2.9 mm Hg and 11.6±3.8 mm Hg, respectively. The cases with open or just partly ligated vertical vein in three groups were 0 case （0%）, 7 cases (14.3%) and 2 cases (11.8%), respectively. The cases of arrhythmia in three groups were 5 cases （25.0％）, 15 cases （30.6%）and 1 case （5.9%）. The severely low cardiac output syndrome occurred in 2 patients and reoperation for bleeding in 2 patients. The morbidity of arrhythmia in group Ⅲ was less than in group Ⅱ（P=0.042）. Conclusion The outcome of surgical repair for S -TAPVC is satisfactory. The anastomosis through the top of the left atrium incision has low occurrence of arrhythmia. The anastomosis through the right and left atrium incision is easy to expose and to perform surgery, especial for old children and adult patients.
Objective To investigate the long effect of nonpulsatile flow on changes of structure and function in pulmonary microcirculation and to identify the pulmonary reconstruction under this blood perfusion. Methods Canine models with nonpulsatile flow in the right lung was established, and sacrificed 6 months later. Compare endothelial nitric oxide synthase (eNOS) in vascular endothelium, apoptosis in smooth muscle cell with immunohistochemistry by streptavidinbioepidermmultienzyme complex methodes, and observe structural changes in pulmonary arterioles with optical microscope. Results The expression of eNOS in the right nonpulsatile flow perfusing lung was weaker as compared to the left lung （10 846.7±177.8 vs. 13 136.1±189.6;t＝2.240, P=0.040）, the fas was ber as compared to the left lung（14 254.1±217.1 vs. 11 976.7±195.7; t＝2.160, P=0.040）. The ratio of wall thichness/vessel diameter in the right lung（13.64%±12.80% vs. 14.96%±13.10%） and wall area/vessel area(46.40%±11.70% vs. 47.80%±12.20%) was lower as compared to the left lung(Plt;0.05). Conclusion Longterm nonpulsatile flow can decrease the expression of eNOS, contract the muscles in capillary net, and increase pulmonary vascular resistance. Moreover it canincrease the arteriole apoptosis, leading to vascular structure remodeling.