Abstract： Objective To construct a nesprin-siRNA lentiviral vector（LV-siNesprin）, transfect it into bone marrow mesenchymal stem cells （MSCs）, and observe morphology changes of MSCs. Methods According to the target gene sequence of nesprin, we designed and synthesized four pairs of miRNA oligo, which were then annealed into double-strand DNA and identified by sequencing. MiRNA interference with the four kinds of plasmids （SR-1,SR-2,SR-3, andSR-4） were transfected into rat vascular smooth muscle cells, and reverse transcriptase chain reaction(RT-PCR) and Western blotting were performed to detect the interference effects and filter out the most effective interference sequence. We used the best interference sequence carriers and pDONR221 to react together to get the entry vectors with interference sequence. Then the objective carrier pLenti6/V5-DEST expressing both entry vectors and lentiviral vectors was restructured to get lentiviral expression vector containing interference sequence （LV-siNesprin＋green fluoresent protein （GFP））, which was packaged and the virus titer was determined. LV-siNesprin＋GFP was transfected to MSCs, and the expression of nesprin protein（LV-siNesprin＋GFP group，GFP control group and normal cell group）was detected by Western blotting. The morphology of MSCs nuclear was observed by 4’,6-diamidino-2-phenylindole （DAPI） stain. The proliferation of MSCs （LV-siNesprin＋GFP group，GFP control group and normal group） was detected by 3-(4,5-dimethylthia- zol-2-yl)-2,5-diphenyltetrazolium bromide（MTT） after lentivirus transfected to MSCs at 24, 48, 72, and 96 hours. Results The four pairs of miRNA oligo were confirmed by sequencing. Successful construction of LV-siNesprin was confirmed by sequencing. The best interference with miRNA plasmid selected by RT-PCR and Western blotting was SR-3. Lentiviral was packaged, and the activity of the virus titer of the concentrated suspension was 1×106 ifu/ml. After MSCs were transfected with LV-siNesprin, nesprin protein expression significantly decreased, and the nuclear morphology also changed including fusion and fragmentation. The proliferation rate of MSCs in the LV-siNesprin＋GFP group was significantly slower than that of the GFP control and normal cell groups by MTT. Conclusion Nesprin protein plays an important role in stabilizing MSCs nuclear membrane, maintaining spatial structure of MSCs nuclear membrane，and facilitating MSCs proliferation.
Objective To investigate clinical outcomes and perioperative management of off-pump coronary artery bypass grafting （OPCAB） for patients following acute myocardial infarction （AMI）.?Methods?From January 2006 to March 2010， 239 consecutive patients underwent OPCAB on the 14-27 （20.55±3.91） d following AMI（AMI group）in Renji Hospital，School of Medicine of Shanghai Jiaotong University. Preoperative MB isoenzyme of creatine kinase（CK-MB） level was （15.82±6.24） U/L and cardiac troponin I（cTnI） was （0.07±0.04） ng/ml. Clinical data of 406 patients without myocardial infarction history who underwent OPCAB during the same period were also collected as the control group for comparison.?Results?The 30-day mortality of AMI group was 2.51% （6/239）. The causes of death were circulatory failure in 4 patients， ischemic necrosis of lower extremity caused by intra-aortic balloon pump （IABP） in 1 patient and pneumonia with septic shock in 1 patient. Dopamine usage in AMI group was significantly higher than that of the control group （61.51% vs. 37.44%， P=0.001）. Intraoperative or postoperative IABP implantation was more common in AMI group， but there was no statistical difference between the two groups（P＞0.05） . Postoperative drainage and blood transfusion in AMI group were significantly larger than those of the control group （385.18±93.22 ml vs. 316.41±70.05 ml， P=0.022；373.68±69.54 ml vs. 289.78±43.33 ml， P=0.005， respectively）. But there was no statistical difference in re-exploration rate between the two groups （P＞0.05）. There was no statistical difference in the incidence of postoperative new onset atrial fibrillation between the two groups （P＞0.05）. Incidence of acute kidneyinjury of AMI group was significantly higher than that of the control group （13.81% vs. 8.62%， P=0.038）. Postoperative 30-day mortality of AMI group was higher than that of the control group， but there was no statistical difference between the two groups （2.51% vs. 1.48%，P＞0.05）. There was no statistical difference in ICU stay time and postoperative hospital stay between the two groups （2.01±0.95 d vs. 1.78±0.98 d；10.33±4.16 d vs. 9.89±4.52 d， respectively， P＞0.05）. A total of 211 patients （88.28%）in AMI group were followed up for 2.89±1.02 years， and 28 patients （11.72%） were lost during follow-up. Twenty-five patients died during follow-up including 14 cardiac deaths. One-year survival rate was 97.63%， and five-year survival rate was 88.15%.?Conclusion?It’s comparatively safe to perform OPCAB for patients at 2-4 weeks following AMI when their CK-MB and cTnI levels have returned to normal range.
Abstract： Objective To evaluate the incidence and prognosis of postoperative acute kidney injury （AKI） in patients after cardiovascular surgery， and analyse the value of AKI criteria and classification using the Acute Kidney Injury Network （AKIN） definition to predict their in-hospital mortality. Methods A total of 1 056 adult patients undergoing cardiovascular surgery in Renji Hospital of School of Medicine， Shanghai Jiaotong University from Jan. 2004 to Jun. 2007 were included in this study. AKI criteria and classification under AKIN definition were used to evaluate the incidence and in-hospital mortality of AKI patients. Univariate and multivariate analyses were used to evaluate preoperative， intraoperative， and postoperative risk factors related to AKI. Results Among the 1 056 patients， 328 patients（31.06%） had AKI. In-hospital mortality of AKI patients was significantly higher than that of non-AKI patients （11.59% vs. 0.69%， P＜0.05）. Multivariate logistic regression analysis suggested that advanced age （OR=1.40 per decade）， preoperative hyperuricemia（OR=1.97）， preoperative left ventricular failure （OR=2.53）， combined CABG and valvular surgery （OR=2.79）， prolonged operation time （OR=1.43 per hour）， postoperative hypovolemia （OR=11.08） were independent risk factors of AKI after cardiovascular surgery. The area under the ROC curve of AKIN classification to predict in-hospital mortality was 0.865 （95% CI 0.801-0.929）. Conclusion Higher AKIN classification is related to higher in-hospital mortality after cardiovascular surgery. Advanced age， preoperative hyperuricemia， preoperative left ventricular failure， combined CABG and valvular surgery， prolonged operation time， postoperative hypovolemia are independent risk factors of AKI after cardiovascular surgery. AKIN classification can effectively predict in-hospital mortality in patients after cardiovascular surgery， which provides evidence to take effective preventive and interventive measures for high-risk patients as early as possible.