Biomarkers ; blood ; metabolism ; Diabetes Mellitus, Type 2 ; blood ; diagnosis ; genetics ; metabolism ; Humans ; MicroRNAs ; genetics ; Polymorphism, Single Nucleotide ; genetics ; Precision Medicine ; methods

Objective The feasibility of constructing endothelialized vascular graft by using cryopreserved HUVCs was studies. Methods Forty-two human umbilical cords were used in this study. HUVECs were isolated by means of filling umbilical veins with digestive enzyme solution. HUVECs were then cultured and observed. Endothelial cells were identified by von Willebrand factor immunofluorescence staining and scanning electron microscope examination. Endothelial cells were suspended in cryopreserving solution which contains 10% DMSO and 10% fetal bovine serum in M199 and cryopreserved in liquid nitrogen. Post-thawed cells and non-frozen cells proliferation was evaluated by measuring the metabolic activity of tetrazolium compound. The endothelial cell growth characteristics were determined by daily observation using phase contrast microscope. Post-thawed endothelial cells viability was determined by trypan blue staining test. Flow cytometry were applied to determine the apoptosis rate of post-thawed cells. Cryopreserved endothelial cells morphological examinations such as hematoxylin and eosins staining and scanning electron microscope examination were carried out in this study. After cell culture and amplification, cryopreserved HUVCs were seeded on the inner surface of Gore-Tex graft to construct tissue engineered vascular graft. Results Extreme high-purified endothelial cells could be isolated by infusing digestive solutions to the lumen of human umbilical veins. Compared with non-frozen endothelial cells, Post-thawed endothelial cells showed 95% of vitality. Post-thawed HUVEC growth curve was similar to non-frozen ones'. Post-thawed HUVEC apoptosis rate (5.85? 0.56) % was higher than non-frozen ones (5.34?0.49)%; however, the difference was not statistically different. Endothelialization of vascular graft was carried out successfully. Cryopreserved cells on Gore-Tex surface showed a good growth trend. Conclusion Cryopreserved HUVCs may be taken as a cell choice for tissue engineering. Enough high pure endothelial cells could be isolated by digestive solutions infusion of human umbilical veins. Post-thawed endothelial cells are proved to have high vitality and growth potential on Gore-Tex surface in vitro.

Objective To evaluate the outcomes of cardiac surgery in neonates weighing less than 2500g and premature infants. Methods From January 2000 through March 2005, 36 consecutive critically symptomatic neonates (26 males, 10 females) weighing less than 2500g underwent cardiac surgery at our center. Median gestational age was 34 weeks with 24 (70.6%) premature (≤37 weeks). Median age at operation was 15.5 days (from 4 days to 68 days). Median weight was 2120g (from 700g to 2500g). Indications for surgery were: persistent ductus arteriosus (PDA, n=15), coarctation of the aorta (CoA, n=3), transposition of great arteries with severely hypoplastic aortic arch (TGA/HAA, n=1), ventricular septal defect (VSD, n=10), pulmonary atresia with intact ventricular septum (PA/IVS, n=5), total abnormal pulmonary venous return (TAPVR, n=2). The heart functions of all patients were assessed in NYHA class IV and 7 patients (19.4%) were intubated pre-operatively. 18 cases (50%) needed extracorporeal circulation and deep hypothermia with circulatory arrest was used in 6 patients. Results The overall mortality was 11.1%. The causes of death were left heart failure in 1 patient and multiorgan failure in 2 and hypoxmia in 1. Mean extracorporeal circulation time was 92 min (from 72 min to 198 min). Mean aortic cross-clamping time was 76 min (from 46 min to 158 min). Mean deep hypothermia with circulatory arrest Peritoneum dialysis was carried out in 5 cases. Mean follow-up period was 22. 3 months (from 2 months to 46 months). 2 patients underwent reoperation and catheter intervention respectively. The heart functions of 26 patients (81.4%) were in NYHA class I at the latest clinical examination. Conclusion This study suggests that complete repair of simple and complex congenital heart diseases can be performed effectively in low birth weight neonates with acceptable mortality and morbidity. Low weight alone should not be considered as a contraindication to surgery in these patients. It is recommended that such neonates undergo early surgical correction rather than prolonged conservative palliation.

Objective To evaluate the expression and the relativity of PMS2,Akt1 and P-AktS473 protein in A2780,Caov3,C13* and ES2 ovarian cancer cell lines .Methods The expression of PMS2 ,Akt1 and P-Akt S473 protein in A2780 ,Caov3 ,C13*and ES2 ovarian cancer cells was detected by Western Blot .After treated with IGF-1 (Akt1 activator) and API-2 (specific Akt1 in-hibitor) ,Caov3 ,ES2 and A2780 cells were collected and the level of PMS2 was detected by Western Blot .Results PMS2 ,Akt1 and P-Akt S473 proteins were detected in all of the four ovarian cancer cell lines with varied expression levels ,and the activity of Akt1 was inversely related to PMS2 expression in ovarian cancer cells .Exposed to Akt kinase stimulator IGF-1 ,ES2 and Caov3 cells were detected with a dramatically PMS2 decreasing .Meanwhile ,the decreasing of PMS2 protein was time-dependent on IGF-1 .Treated with API-2 ,Akt kinase specific inhibitor ,A2780 was detected with PMS2 dramatically increasing ,and the increasing of PMS2 pro-tein was time-dependent on API-2 .Conclusion In ovarian cancer cells ,PMS2 expression could be directly regulated by activated Akt1 .

Objective To evaluate the clinical value of real-time 3 dimensional echocardiography (RT-3DE) for measurement of right ventricular function in the repaired tetralogy of Fallot (TOF) compared to cine magnetic resonance imaging (c-MRI).Methods Thirty-seven randomly selected patients with repaired TOF from Jan.2000 to Jun.2007 in Children's Hospital,Fudan University underwent both RT-3DE and c-MRI for the data of right ventricular enddiastolic volume (RVEDV),right ventricular end-systolic volume (RVESV),right ventricular ejection fraction (RVEF),which were compared to evaluate the reproducibility and correlation of two ways on right ventricular function.Results All the patients completed both RT-3DE and c-MRI on the same day.The correlation for RVEDV,RVESV,RVEF measured by 2 methods was high (r =0.933,0.943,0.911,P =0.000,0.000,0.105),and it showed that consistency existed in RVEF,as difference was only (1.2 ± 4.5) ％.Comparison with c-MRI,RVEDV and RVESV measured by RT-3DE was underestimated,in which difference was (-17.4 ± 16.9) mL,(-9.3 ± 10.5) mL.Conclusions RVEF with RT-3DE can be assessed with acceptable accuracy,but it is not so good for RVEDV and RVESV.Further study needs to be performed to make sure the value of RT-3DE on the assessment of right ventricular function in repaired TOF.

Objective To evaluate early and mid-term results in infants,weighing less than 2 500 g,who underwent early cardiac surgery on cardiopulmonary bypass (CPB).Method Since November 2003 to December 2007,28 consecutive infants of less than 2 500 g underwent early cardiac surgery on CPB in Children's Hospital Affiliated to Fudan University.Sixteen infants were male,and 12 infants were female.The mean pregnancy period was 34 weeks (30 to 41 weeks) ,and 14 patients were premature,and the mean birth weight was 2 010 g(1 370-2 500 g).Mean age at operation was 27.46 d (range 1～61 d),and mean weight was 2 391 g (range 1 500～2 500 g).Deep hypothermia circulatory arrest was used in 9.Indications for surgery were:ventricular septal defect (10 infants),transposition of great arteries (6 iofants),totally anomalous pulmonary venom return (5 infants),pulmonary atresia with intact ventricular septum(2 infants),coarctation of aorta (1 infants) ,interrupted aortic arch (1 infants),cortriatriatum(1 infants),abnormal original of fight pulmonary artery (1 infants).Resuits Two (7.14%) infants died after operation at early phase.Mean duration of mechanical ventilation and intensive care unit stay were 113.9 h and 14.4 d,respectively.During follow-up (mean 16.87 months),one died.Two patients needed reintervention.About 84.4% infants didn' t need reintervention at the end of 12-month followup.Conclusions If medicine is aseless,infants woth low birth weight and congenital heart defect can obtain satisfactory outcome after CPB.Perioperative proration for virtual organs should be payed attention to.

Objective　To investigate the prognosis in risk of ventricular arrhythmia in coronary heart disease with heart rate variability (H RV), left ventricular ejection fraction (LVEF) and other clinical background dat a. Methods　A total of 81 patients were divided into ventricular premature beats (VPBs)≥30/h group and VPBs<30/h group. Their LVEF, HRV and cli nical data were studied and analyzed. Results　The age and blood pressure between 2 groups had no significant difference. LVEF, standard deviati on of all normal RR intervals (SDNN), SD of the average of NN interval (SDANN) a nd HRV triangular index (HRVI) were significant less in VPBs≥30/h group than in VPBs<30/h group (43.29±15.38 vs 67.33±11.47，P＜0.01；90.05±22.2 9 vs 117.90±30.32，P＜0.05；77.43±17.78 vs 105.69±28.79，P＜0.05 ；24.54±8.70 vs 32.70±10.87，P＜0.05, respectively). Incidence of myo cardial infarction (MI) was larger in VPBs≥30/h group than VPBs<30/h group. LVE F was the independent predictable factor in risk of ventricular arrhythmia with multinomial regression logistic analysis（B=0.119, P=0.032）. Co nclusion 　Our findings indicate that LVEF is an independent predictable factor i n risk of ventricular arrhythmia in coronary heart disease. Although HRV and MI history can not be used to predict VPB, significant difference is found between 2 groups. High-risk patients could be selected successfully when these data are considered in combination.

At present, microbe surface display system mainly involves phage surface display system, bacte- rial surface display system, yeast surface display system and virus surface display system. Baculovirus sur- face display system is a new type of eukaryote surface display system which developed based on deeply un- derstanding of construction and function of virus genome in recent years. Through fused expression with vi- ral capsid or membrane proteins exogenous peptides can be displayed on the surface of the virus and formed hedgehog-shape "fake virus". Baculovirus surface display system was characterized by safeness and high performance, furthermore, this system can complete post-translation processing and modification of protein to enhance the bioactivity of exogenous product. Combined with the author’s experimental work, this paper briefly introduces the mechanism and traits of this system and summarizes the newest research development on its application in the field of monoclonal antibody preparation, new-type vaccine development, genes transduction and genes therapy. It is believed that the system above may show extensive application through further improvement and optimization.

Objective To construct the eukaryotic expressing plasmid of tumor necrosis factor-related apoptosis-inducing ligand(TRAIL),and study its inhibitory effect on hepatic tumor which implanted subcutaneously in nude BALB/c mice.Methods Total RNA of U937 cell was extracted, and its extracellular domain (114-281aa) was amplified by RT-PCR, then signal peptide was ligated. The recombinant secreting plasmid for TRAIL was constructed successfully which was confirmed by enzyme cleavage identification and sequencing identification. Liver cancer cell (strain No.7402) was implanted subcutaneously in 32 nude BALB/c mice. These mice were randomly divided into two groups: study group and control group. The mice in study group received muscular injection of plasmids for transfection, and the mice in control group received the injection of normal saline at the same time. The size of implanted tumors were measured continuously till the day of sacrificing, tumor cell apoptosis effect was examined by TUNEL method. Results In study group,tumor volume was smaller than that in control group and the blue-purple apoptosis cells were observed under microscope. Conclusion TRAIL plasmid can induce apoptosis of liver cancer cell and can inhibit the growth of liver tumor.

Corneal endothelial cell(CEC)is the most critical part for the cornea, of which activity can influence the postoperative vision.It is very important for the clinical cornea preservation considering the function and its self-purification of donor cornea.There are a variety of classical methods, which can significantly prolong the saving time of donor cornea with its good quality of CEC.We reviewed the published papers about present preservation methods of cornea, which can give us many suggestions for the clinical cornea preservation.