BACKGROUND: Hyperpolarization-activated cyclic nucleotide-gated (HCN) current plays an important role in regulating heart spontaneous pulsation.OBJECTIVE: To observe target gene expression and electrophysiological characteristics of pig bone marrow mesenchymal stem cells (BMSCs) transfected with hyperpolarization-activated cyclic nucleotide-gated channel 2 (HCN2) gene recombinant adenovirus.DESIGN, TIME AND SETTING: Cell-gene in vitro study was performed at the Laboratory of Thoracic and Cardiovascular Surgery,Second Military Medical University of Chinese PLA from July 2007 to March 2008.MATERIALS: Yorkshire pig was supplied by Animal Institute, Second Military Medical University of Chinese PLA. HCN2 plasmid was presented by Professor Dario DiFrancesco from Italy. Recombinant adenovirus Ad.HCN2 was constructed and stored using Ad5 in this laboratory.METHODS: Pig BMSCs were isolated with combination of gradient centrifugation of Percoll and adherent treatment in vitro.Ad.HCN2 was transfected at multiplicity of infection=50. We also set non-transfection and transfected Ad.Null groups.MAIN OUTCOME MEASURES: Expression of HCN2 mRNA was detected with RT-PCR, and expression of HCN2 channel protein was examined with immunofluorescent staining. Electrophysiology of HCN2 channel protein was measured with whole-cell patch clamp.RESULTS: No amplified fragments were found in the non-transfection and transfected Ad.Null groups, but amplified fragments were determined at 250-500 bp following Ad.HCN2 amplification, which was the same as plasmid carrying HCN2 gene. Staining strength of cell nuclei following transfection was significantly weaken compared with cell membrane and plasma, which showed identical distribution as HCN2 protein. No HCN2 protein was detected in the non-transfection and transfected Ad.Null groups.Pacemaker current could be recorded with a whole-cell patch clamp. It was fully activated around -140 mV with an activation threshold of -60 mV, presenting voltage dependence. CsCI (4 mmol/L) reversibly blocked the inward currents. No pacemaker current was detected in the non-transfection and transfected Ad.Null groups.CONCLUSION: The HCN2 recombinant adenovirus carrier was transferred into serial subcultivation porcine bone marrow mesenchymal stem cells. HCN2 channel protein has been expressing. Pacemaker current could be recorded with a whole-cell patch clamp.

Objectives:To study the postoperative nutritional support in patients with pancreatic disease. Methods:Based on the kind of pancreatic disease,twenty two patients were randomly divided into PN+EN group and TPN group during 20 days after operation.Body weight,the restoring time of intestinal movement,the complications,the blood amylase,the serum glutamic pyruvic transaminase,the blood urea nitrogen,the blood sugar,the serum albumin(Alb),transferritin(TFN) and prealbumin(PAB) were compared on the first,fifth,tenth and twentieth day after operation. Results:Body weight of two groups reduced lightly.The blood amylase decreased remarkably.The functions of liver and kidney were recovered gradually.The blood sugar tended to restore normal.Alb,TFN,PAB of two groups increased remarkably( P

ing microecological preparation,or continuously pumping EN fluid into the stomach or jejunum may reduce the incidence of diarrhea.

Objective To develop a novel method for treating complete heart block by autotransplantation of simus node node cells to right ventricular anterior wall.Methods Twenty healthy mongrel dogs were involved in the present study.The dogs were randomly assigned to transplant group or control group(n=10).The sinus node (SN)was harvested and isolated in vitro after an electronic pacemaker was implanted and complete heart bolck was introduced.The SN cells from dogs of transplant group were injected to autogenic right ventricular wall.Commensurable culture medium was implanted to ghe same position of dogs in control group.Two weeks later,detailed electropohysiological study was performed.For investigating the variation of the rhythm,epinephrine was administrated through femoral vein to dogs of transplant group.Results Most of isolated SN cells from dogs were thin-spindle shape,and cell activity was fine.The SNs by VG stained displayed typical structural feature.2 weeks after cell autotransplantation,higher heart rates were achieved from transplant group than that in control group(P＜0.05).This rhythm was stable in 4 weeks and became faster remarkably after administration of eninephrine(P＜0.05).Conclusion SN cell of dogs tutorgrfted into right ventricular anterior wall can form new pacemaker site in ventrcle and improve ventricular rate of complete heart bolck.This pacemaker site can also be regulated by epinephrine.

Objective To evaluate the effect of proper nutritional therapy guided by indirect caloimetry . Methods According to the digitaltable ,70 critically ill patients whose APACHE II >5 were randomly divided into oberserving ( nutritional therapy guided by indirect caloimetry ) and control groups ( nutritional therapy guided by harris-benedict formula),each group 35 cases.The results of nutrition support were analysed .Results TP,MAMC,ALB were higher than that before treatment in oberserving group (all P<0.05),the MAMC was also higher than that in control groups(P<0.05).There was no significant difference in Hb and TSF after treatment IN oberserving group (all P>0.05).Conclusion The nutritional therapy guided by indirect caloimetry by harris-benedict formula have the some nutritional effect in critically ill patients .

Objective To study the beneficial effect and mechanism of Alcea rosea roots in nephrolithiasis model in-duced by 1%ethylene glycol in rats. Methods We randomly divided 60 male Wistar rats into six groups,including control group, model group, Alcea rosea roots lower-dose preventive group, Alcea rosea roots high-dose preventive group, Alcea ro-sea roots lower-dose curative group, Alcea rosea roots high-dose curative group. Control group was free to access food and water;model group was given 1%ethylene glycol drinking water and was fed with normal diet, preventive group was given 1%ethylene glycol drink and Alcea rosea roots in low (250 mg/kg) or high dose (500 mg/kg) each day, curative group re-ceived 1%ethylene glycol drink each day and Alcea rosea roots in low or high dose from day 15 to day 28. At the end of the experiment, various renal functional and injury markers such as urine volume, calcium, phosphate, magnesium, urea, creati-nine, and oxalate were evaluated using urine, serum, and kidney homogenates. The kidneys were removed and prepared for histological evaluation of calcium oxalate deposits. Results In model groups, urine output, urea, creatinine, 24 h urine Ca2+, and oxalate and MDA were increased compared to those in control group(P＜0.05). GSH and SOD were increased in preventative and curative groups compared to those in the model group(P ＜ 0.05). The urea, creatinine, 24 h urine Ca2+, urine oxalate, MDA were reduced in preventive and curative groups compared to those in the model group(P＜0.05). The number and size of calcium oxalate crystal deposits were also less and smaller, and the kidney damage was less severe in pre-ventive and curative groups compared to in the model group. Conclusion The extract of Alcea rosea roots can prevent and treat calcium oxalate urinary stone formation in rats and protect renal function.

Objective To explore the distribution of lactate and base excess in children with Tetralogy of Fallot in perio-perative period, its dynamic variation and its relationship with prognosis.Methods Perioperative arterial lactate level was dy-namically monitored in 1 552 children with tetralogy of Fallot.According to the prognosis, 1 505 subjects were assigned to sur-vive group, and 47 subjects to nonsurvive group.The lactate and base excess levels of all subjects were collected after induction (T1), after protamine administration(T2), and 24 h(T3) after surgery in the intensive care unit(ICU).Results The pa-tients who could not survive showed significant elevated lactate levels as compared to patients who survived after surgery [(7.91 ±5.76)mmol/L vs.(1.20 ±0.51)mmol/L, P＜0.05].However, in nonsurvivors, the BE value decreased signifi-cantly in the ICU period in comparison to survivors[(－1.91 ±5.09)mmol/L vs.(2.52 ±2.85)mmol/L, P＜0.05].Logis-tic regression analysis was performed, indicating the elevated lactate and depressed base excess levels measured 24 h after sur-gery were independent risk factors.Conclusion The dynamic monitoring of lactate and base excess levels in the perioperative period in patients with tetralogy of Fallot were sensitive indicators to assess severity , to guide treatments, and for the prognosis. In the perioperative period, when the lactate increased and(or) base excess levels decreased, the mortality rate increased as well.lactate and base excess levels measured 24 h after surgery in the ICU were independent risk factors that significantly in-creased the mortality rate.

Objective The study was designed to analyze the risk factors for perioperative adverse events in Ebstein's a-nomaly(EA) after cone construction.Methods A retrospective analysis was performed on EA cases undergoing cone con-struction in pediatric heart center of Beijing Anzhen Hospita from January 2010 to January 2017.Patients were divided into group A( no adverse events occurred) and group B( adverse events occurred) according to the presence or not of perioperative adverse events.Univariate and multivariate logistic regression analysis was performed with 9 preoperative and intraoperative var-iables as the predicted risk factors.Results The incidence of adverse events was 13.1％.Univariate logistic regression analy-sis showed that preoperative percutaneous oxygen saturation(SpO2)≤0.90(P=0.001, OR=10.578) and extracorporeal cir-culation time(P=0.034, OR=1.021) were risk factors for perioperative adverse events.Results of multivariate logistic re-gression analysis: preoperative SpO2≤0.90 was an independent risk factor for perioperative adverse events(P=0.002, OR=9.108).In 64 cases(64.6％) with atrial septum defect, there was no significant correlation between preoperative SpO2 and preoperative tricuspid regurgitation(P=0.838, r =0.026).Conclusion Preoperative SpO2≤0.90 is an independent risk factor for adverse events in the perioperative period of cone reconstruction .It is a simple and efficient method to evaluate the operation difficulty and determine the prognosis by SpO 2 .

Asian Continental Ancestry Group ; Hearing Loss ; genetics ; Humans ; Mutation ; genetics ; Pedigree ; RNA, Ribosomal ; genetics