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 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.
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 effect of ischemic preconditioning(IPC) on myocardial ischemic reperfusion injury(I/R) of elderly rats. Methods Fiftysix Wistar rats, of which there were 28 aged from 21-23months(elderly rat) and 28 aged from 4-5months(young rat), were used to build isolated heart perfusion Langendorff model. The rats were divided into 7 groups with random number table（8 in each group）: adult control group, adult I/R group, adult IPC group, elderly control group, elderly I/R group, elderly IPC group and elderly enhanced IPC group. The control group underwent a 90-min perfusion without any intervention; the I/R group underwent a 30-min equilibration period, then a 30-min ischemia and a 30-min reperfusion; the IPC group underwent a 10-min equilibration period, then a 5-min ischemia for twice and a 5-min reperfusion, after that a 30-min ischemia and [CM(158-3mm]a 30-min reperfusion; the enhanced IPC group underwent a 10-min equilibration period, then a 5-min ischemia for 4-times and a 5-min reperfusion, after that a 30-min ischemia and a 30-min reperfusion. The recovery rates of cardiac output(CO), left ventricular developed pressure (LVDP), the maximum rising and descending rate of left ventricular pressure (±dp/dtmax) after a 30-min reperfusion were compared among groups. The activity of creatine kinase (CK) in coronary outflow, the level of malonyldialdehyde (MDA) and superoxide dismutase (SOD) before ischemia and after a 30min reperfusion were detected. The myocardial infarction areas were compared among groups. Results After a 30min reperfusion, compared with adult I/R group, in adult IPC group CK reduced significantly(89.48±18.72 U/L vs. 115.76±16.72 U/L，q=6.061，Plt;0.01)，the level of MDA decreased significantly（9.53±3.44 nmol/ml vs. 16.84±2.29 nmol/ml，q=7.732，Plt;0.01)，the level of SOD increased significantly（584.7±122.62 U/ml vs. 429.46±85.24 U/ml，q=4.754，Plt;0.01)，the recovery rates of CO,LVDP,+dp/dtmax and -dp/dtmax increased ignificantly(78.69%±9.68% vs. 65.10%±8.63%，83.61%±8.46% vs. 67.23±8.68%，81.68±8.68% vs. 67.89%±6.89%，89.79%±7.78% vs. 66.79%±8.46%，Plt;0.01), the myocardial infarction areas reduced significantly (5.25%±4.33% vs. 14.75%±8.02%,q=7.458，Plt;0.01)。There was no statistical significance between elderly IPC group and elderly I/R group in the above indexes（Pgt;0.05）.However, There was statistical significances between elderly enhanced IPC group and I/R group. CK reduced significantly (88.60±28.32 U/L vs. 105.76±9.64 U/L，q=5.620，Plt;0.01)，the level of MDA decreased significantly（8.38±3.36 nmol/ml vs. 16.80±3.06 nmol/ml，q=7.500，Plt;0.01)，the level of SOD increased significantly（558.87±78.66 U/ml vs. 433.75±86.65 U/ml，q=7.335，Plt;0.01)，the recovery rates of CO,LVDP,+dp/dtmax and -dp/dtmax increased significantly (77.99%±10.02% vs. 66.26%±9.78%，85.59%±6.67% vs. 73.90%±6.66%，83.87%±9.98% vs. 68.90%±8.68%，86.01%±766% vs. 70.39%±7.98%，Plt;0.01)， the myocardial infarction areas reduced significantly (795%±6.32% vs. 1568%±10.36%,q=8.680， Plt;0.01). 〖WTHZ〗Conclusion The protective effect of IPC on I/R elderly rat hearts has weakened. The enhanced IPC is able to regain the protective effect of IPC on elderly rat hearts.
Abstract: Objective To evaluate the outcome of reconstruction of right ventricular outlet tract (RVOT) with 0.1 mm Gore-Tex monocusp valve for short and middle term. Methods Between June 2002 to July 2006, 48 patients underwent reconstruction of RVOT with Gore-Tex monocusp valve to correct cardiac anomalies, including 33 patients with tetralogy of Fallot (TOF) and pulmonary stenosis, 8 patients with TOF and pulmonary atresia, 3 patients with TOF and absent pulmonary valve, 2 patients with double outlet of right ventricle and pulmonary stenosis, 1 patient with truncus arterious and 1 patient with complete transposition of great artery, ventricular septal defect and pulmonary stenosis. Results There was no operative death. The postoperative blood oxygen saturation was up to 1.00. The ratioes of right ventricular systolic pressure and left ventricular systolic pressure were between 0.22 to 0.65.The gradient between right ventricle and left or right pulmonary artery was less than 10 mmHg. All patients were followed up including echocardiography ranged from 3 to 48 months. There were no late death and complication. Trivial and mild pulmonary insufficiency was detected in 18 patients and valvular motion remained competent in 40 patients. Conclusion The results suggest that the reconstruction of RVOT with Gore-Tex monocusp valve can achieve excellent outcome for short and middle term.
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.
Abstract： Objective?To analyze the risk factors for early shunt dysfunction after systemic-pulmonary shunt in order to improve early postoperative outcomes. Methods We retrospectively analyzed the clinical records of 189 patients who underwent systemic-pulmonary shunt in General Hospital of Shenyang Military District between February 2002 and December 2010. There were 87 males and 102 females with their age ranging from 3 months to 50（5.3±6.2）years，and body weight ranging from 3 to 56（17.7±11.0）kg. There were 94 patients with tetralogy of Fallot （TOF） and pulmonary artery stenosis，51 patients with pulmonary atresia （PA） and ventricular septal defect，4 patients with PA and intact ventricular septum，10 patients with functional single ventricle （SV） and pulmonary stenosis（PS），6 patients with SV and PA，6 patients with double outlet right ventricle and PS，8 patients with transposition of the great arteries（TGA） and PS，and 10 patients with TGA and PA. The surgical procedures included central aorto-pulmonary shunt （Waterston） in 105 patients, modified Blalock-Taussig shunt in 61 patients and Melbourne shunt in 23 patients. Results Early postoperative death occurred in 13 patients （6.9%）. There were 12 patients （6.3%） with intra-operative severe hypotension or arrhythmia, 10 patients （5.3%） with postoperative severe low cardiac output, and 10 patients （5.3%） with early shunt dysfunction within the first 24 h postoperatively. Univariate analysis identified low body weight （P=0.027）, shunt size with diameter＜4 mm （P=0.025） and severe intra-operative adverse event （hypotension or arrhythmia with P=0.002） were risk factors for early shunt dysfunction. In multivariate analysis, intra-operative adverse event was an independent risk factor for early shunt dysfunction（P=0.017）. Conclusion Early outcomes after systemic-pulmonary shunt can be significantly improved by preventing intra-operative severe hypotension or arrhythmia and choosing shunt size larger than 4 mm，especially for patients with young age, low body weight, and poorly-developed pulmonary artery.
Abstract： Objective To investigate the cause, treatment， and prevention strategy of early left ventricular rupture after mitral valve replacement （MVR）. Methods We retrospectively analyzed clinical data of 7 patients who had early left ventricular rupture after MVR， among a total of 2 638 MVR patients， between May 1981 and November 2010 in General Hospital of Shenyang Military District. There were 2 male patients and 5 female patients with their age ranging from 28 to 71 years old. One patient was New York Heart Association （NYHA） functional classⅡand 6 patients were NYHA functional classⅢ. Preoperative echocardiography showed that their left ventricular ejection fraction （LVEF） was 49%-60% and their left ventricular end diastolic volume （LVEDV） was 29-42 ml. All the patients underwent prothetic valve replacement under moderate hypothermia， general anesthesia and extracorporeal circulation. Results All the 7patients underwent surgical repair for their early left ventricular rupture. There were 2 patients with TypeⅠ， 4 patients with TypeⅡ， and 1 patient with TypeⅢleft ventricular rupture. Four patients died of hemorrhagic shock or low cardiac output syndrome. Three patients were successfully healed and discharged from hospital 7-15 days after the operation. The 3 surviving patients were followed up for 4.5-18.0 years, and they all had good general condition and satisfactory quality of life. No late pseudo-aneurysm was found during follow-up. Conclusions Early left ventricular rupture is difficult to repair with a high mortality. Effective prevention strategy should be applied to avoid it as much as possible. Once left ventricular rupture occurs during the surgery， extracorporeal circulation should be quickly established， and it’s possible to save patients’ life by reliable intracardiac and epicardial repair according to patients’ individual condition.