Objective To construct an eukaryotic expression vector with human adipose most abundant gene transcript 1 (APM1) gene,and to investigate the transfection and expression of pCDEF-APM1 eukaryotic expression plasmid in HEK293 cells.Methods pCDEF-APM1 eukaryotic expression plasmid was constructed by DNA recombinant method.Expression vector pCDEF-APM1 was transfected into HEK293 cells with Effectene reagent.The level of human adiponectin protein in the supernatant of cell culture media was detected with double antibody sandwich ELISA.Results The sequence of DNA fragment from constructed pCDEF-APM1 plasmid was identical to that published in GenBank.There was raised human adiponectin protein level in culture supernatant of HEK293 cells tnmsfected with pCDEF-APM1.Conclusion The pCDEF-APM1,an eukaryotic expression plasmid for APM1 gene is successfully constructed.High protein expression of adiponectin can be obtained in HEK293 cells transfected with pCDEF-APM1 eukaryotic expression plasmid.

OBJECTIVETo study the renal protective effect of sodium ferulate (SF) and its mechanism in rats with diabetic mellitus (DM).

METHODSDM rats induced by streptozotocin were treated with SF 110 mg/kg per day for 8 weeks. The ratio of kidney weight/body weight (KW/BW), serum triglyceride (TG) and total cholesterol (TC), creatinine clearance rate (Ccr), urinary protein/24 hrs, levels of endothelin-1 (ET-1) and nitric oxide (NO) in renal cortex in rats were measured, the pathological change of kidney were observed and the expression of transforming growth factor-beta 1 (TGF-beta 1) and collagen IV (C-IV) in kidney were examined using immunohistochemical assay. The data obtained were compared with those obtained from untreated DM rats and normal rats respectively.

RESULTSCompared with the normal rats, in DM rats, Ccr, urinary protein/24 hrs, ET-1, expressions of TGF-beta 1 and C-IV were significantly increased in DM model rats (all P < 0.01), and significantly abnormal pathological change in kidney was found. While in the SF treated DM rats, the above-mentioned abnormal changes were all significantly improved.

CONCLUSIONSF has effect in protecting kidney of DM rats, the mechanism might be related with its actions of reducing ET-1 production in kidney and inhibiting the expressions of TGF-beta 1 and C-IV.

Animals ; Collagen Type IV ; metabolism ; Coumaric Acids ; pharmacology ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; Diabetic Nephropathies ; prevention & control ; Endothelin-1 ; metabolism ; Kidney ; metabolism ; pathology ; Male ; Protective Agents ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar ; Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1

Objective To observe the effect of carvedilol on the expression of Cx43 in rabbits with myocardial infarction and its association with ventricular arrhythmia.Method Thirty-six rabbits were randomly divided into three groups in equal number(n=12),namely,myocardial infarction(MI)group,carvedilol group and sham MI group.Rabbits of carvedilol group were administered with carvedilol 5 mg kg~(-1)?d~(-1)after MI induced,while no carvedilol was administered to the MI group.The following observations were made:(1)Cx43 density of the epicardial border zone measured by quantitative immnuoconfocal laser scanning,and(2)cx43 protein expression analyzed by western blot.Results(1)Under immunoconfocal laser microscope,the relative density of Cx43 was(0.16?0.06)% in the infarction group and was(0.32?0.11)% in the sham MI group [(0.68?0.15)%,both P

OBJECTIVETo evaluate the effects of recombinant adenovirus encoding human apM1 gene on proliferation and nitric oxide synthase (NOS) activity in human umbilical vein endothelial cells (HUVECs).

METHODSProtein expression of apM1 in cell culture supernatant of HUVECs transfected with human Ad-apM1 was detected by double antibody sandwich ELISA. The effect of human adiponectin on cell proliferation was assessed by MTT assay. The total NOS and iNOS expressions were measured by chromatometre.

RESULTSHuman adiponectin protein level and total NOS and eNOS expressions were significant increased and iNOS expression significantly reduced in culture supernatant of HUVECs infected with Ad-apM1 compared to that in control HUVECs. The recombinant adenovirus had no influence on HUVECs growth as determined by MTT assay.

CONCLUSIONSHuman Ad-apM1 can be effectively expressed in HUVECs and do not influence HUVECs growth. Increased total NOS and eNOS expressions and decreased iNOS expression in HUVECs transfected with Ad-apM1 gene suggest a potential role of Ad-apM1 gene transfer for the prevention and treatment of arteriosclerosis.

Adenoviridae ; genetics ; Adiponectin ; genetics ; Cell Proliferation ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Endothelium, Vascular ; cytology ; Gene Expression ; Gene Transfer Techniques ; Humans ; Nitric Oxide Synthase ; metabolism ; Umbilical Veins ; cytology

OBJECTIVETo summarize our experience in adult-to-infant living donor liver transplantation (A-ILDLT) and to analyze the efficacy and complications of A-ILDLT.

METHODSThe clinical data, surgical strategies and complications of 28 adult donors and infantile recipients who underwent A-ILDLT from April 2006 to December 2009 were retrospectively analyzed. These 28 patients (14 boys and 14 girls) aged from 80 days to 11.5 months with body weights of 3.08 to 10.3 kg at the time of operation . They suffered from biliary atresia with decompensated cirrhosis. The living donors were 15 mothers, 9 fathers, 3 grandma and 1 elder brother with ABO compatible with the infantile recipients. 27 Donor organs were the left lateral lobe grafts (segment II, III) and 1 graft was segment II. All patients were followed up for 5 to 24 months.

RESULTSThese grafts were orthotopically transplanted into the infantile recipients. The average length of stay was 9.3 days for the donor group without any complications. Postoperative immunosuppression included prednisone, Cyclosporin and mycophenolate mofetil (MMF). A total of 24 postoperative complications occurred in 20 recipients, including 5 vascular complications, 4 bleeding, 7 pneumonia, 2 bowel obstruction, 4 intestinal perforation and 3 rejection. Three recipients died of hepatic arterial thrombosis (HAT). The perioperative mortality rate of recipients was 10.7% (3/28) and the survival rate was 89.3% in peroperative period. One died of stricture of hepatic vein and 1 of accidental asphyxia during follow-up term. At present, 23 cases are still alive.

CONCLUSIONA-ILDLT has become an effective method to infants with end-stage liver disease. The postoperative vascular complication is the predominant cause of death.

Female ; Humans ; Infant ; Liver Diseases ; surgery ; Liver Transplantation ; methods ; Living Donors ; Male ; Retrospective Studies ; Treatment Outcome

BACKGROUNDAccurate views of the head and neck vessels, tumor angiogenesis and the relationship of tumor and the surrounding blood vessels are especially crucial to carotid body tumor (CBT) patients. The aim of this study was to explore the value of dual-source CT (DSCT) cerebral and carotid angiography in CBT diagnosis.

METHODSDSCT cerebral and carotid angiography was performed on nine patients with CBT. Two-dimensional and three-dimensional reconstruction images were obtained by means of multiple planar reconstructions (MPR), maximum intensity projection (MIP) and volume rendering (VR). All patients were subjected to color Doppler flow imaging (CDFI) examination. Two kinds of examinations were performed in 3 days, and all patients were confirmed by surgery.

RESULTSDSCT angiography was successful in all patients. CBTs were diagnosed in 9 patients with 10 lesions (1 case had multiple bilateral CBTs). The largest lesion was 12 cm in diameter, and the smallest one was 1.6 cm in diameter. All patients had clearly demonstrated head and neck vessels, tumor angiogenesis, and tumor relationship with the surrounding blood vessels. The internal and external carotid artery (ICA, ECA) were involved in 2 cases. There were 7 cases with basilar artery ring integrity, and 1 case had the posterior communicating artery absent; 1 case had middle cerebral artery stenosis; 4 cases (4 tumors) showed arterial phase homogeneous enhancement; 5 cases (6 tumors) had obvious heterogeneous enhancement where irregular low-density necrosis could be seen in the tumors. CDFI could demonstrate the nearby blood vessels and tumor structure, instead of tumor angiogenesis. However, DSCT can display both the tumor and the peripheral vascular tumor angiogenesis consistent with surgical findings.

CONCLUSIONSDSCT cerebral and carotid angiography can provide reliable information for the operation. It might be a valuable CBT diagnostic method by showing accurate views of the CBT along with the bilateral neck and brain blood vessels.

Adult ; Carotid Arteries ; diagnostic imaging ; Carotid Body Tumor ; diagnostic imaging ; surgery ; Cerebral Angiography ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods

OBJECTIVETo evaluate the expression of epidermal growth factor receptor (EGFR) gene copy number and the expression of ERCC1 and BRCA1 proteins in patients with non-small-cell lung cancer (NSCLC) and the correlation between them.

METHODSThe status of EGFR gene copy number was determined by in situ hybridization (FISH), and the expression of ERCC1 and BRCC1 proteins was examined by immunohistochemistry (IHC). The relationship of EGFR gene copy number with the expression of ERCC1 and BRCA1 and the clinical pathologic features were analyzed.

RESULTSFISH-positive EGFR expression was identified in 40 of 166 samples (24.1%). More FISH-positive EGFR in the female than male patients (31.9% vs. 18.6%, P = 0.048), and non-smoker than smoker (32.8% vs. 16.7%, P = 0.045). FISH-positive EGFR was not associated with age, pathological type, clinical stage and metestasis status (P > 0.05). The expression of ERCC1 protein was identified in 60 of 132 samples (45.5%). The expression of ERCC1 protein varied significantly in tumors of different pathological types (P = 0.046), but not associated with age, gender, clinical stage, metestatic status and smoking status (P > 0.05). The expression of BRCA1 protein was identified in 46 of 131 samples (35.1%). The expression of BRCA1 was not associated with age gender, pathological type, clinical stage, metestatic ststus and smoking status (P > 0.05). There was a moderate correlation between the expressions of ERCC1 and BRCA1 (r = 0.449, P < 0.001), but EGFR gene copy number was not correlated with the expression of ERCC1 or BRCA1 protein.

CONCLUSIONSFISH-positive EGFR expression is associated with gender and smoking status, but not correlated with the expression of ERCC1 and BRCA1 proteins. There is a moderate correlation between the expressions of ERCC1 and BRCA1.

Adult ; Aged ; Aged, 80 and over ; BRCA1 Protein ; metabolism ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; pathology ; DNA-Binding Proteins ; metabolism ; Endonucleases ; metabolism ; Female ; Gene Dosage ; Gene Expression Regulation, Neoplastic ; Genes, erbB-1 ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Sex Factors ; Smoking ; Young Adult

AIMTo investigate the effect and mechanism of ascorbic acid on podocyte, last barrier of glomerular filtration, in diabetic rats.

METHODSDiabetic rats induced by streptozotocin injection intraperitoneally were treated by ascorbic acid for 5 weeks. The levels of blood glucose (BG), HbA1c, urinary albumin excretion rate (UAER) and superoxide diamutase (SOD), catalase (CAT) and malondialdehyde (MDA) in renal cortex were measured. The podocyte ultrastructure was observed while the expression of desmin protein, a marker of podocyte injury, was examined.

RESULTSCompared with control group, BG and HbA1c were increased markedly in diabetic group. The activities of SOD and CAT were decreased and the concentrations of MDA were increased significantly in diabetic renal cortex. There were the increased proteinic expression of desmin, foot process effacement in podocytes and UAER markedly in diabetic rats. Compared with diabetic rats, foot process effacement and the changes of UAER were ameliorated markedly while the activities of SOD were increased, the levels of MDA and proteinic expression of desmin were decreased markedly although BG, HbA1c and the activities of CAT were no significant difference in the diabetic rats by ascorbic acid treatment.

CONCLUSIONThe findings suggest that there are marked injury in podocyte, last barrier of glomerular filtration, in diabetic rats and administration of ascorbic acid can protect podocyte by increasing antioxidative capacity and ameliorating the renal oxidative stress.

Animals ; Ascorbic Acid ; pharmacology ; Catalase ; metabolism ; Desmin ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; physiopathology ; Diabetic Nephropathies ; metabolism ; pathology ; physiopathology ; Male ; Oxidative Stress ; drug effects ; Podocytes ; drug effects ; metabolism ; ultrastructure ; Random Allocation ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism

OBJECTIVETo observe the prevalence of J wave in apparently healthy subjects in Wuhan.

METHODSThe study subjects comprised of 1817 apparently healthy subjects (1131 males, mean age 46.38 +/- 15.81 years; 686 females, mean age was 42.77 +/- 14.15 years). ECG and routine medical examinations were performed. J wave was defined as a wave followed QRS complexes with amplitude of at least 0.05 mV and 0.03 s.

RESULTSThe overall incidence of J wave was 7.26%. The incidence of J wave in males was significantly higher than in females (10.53% vs. 1.87%, P < 0.01). The incidence of J wave in leads of inferior wall (II, III, avF), right wall (V(1 - 3)) and left wall (V(4 - 6)) was 4.57%, 0.50%, and 2.20%, respectively. J wave located in leads of inferior wall was more than in left and right walls (both P < 0.05). The incidence of J wave positively correlated with age (y = 0.1387x + 1.6318, r = 0.78, P < 0.01).

CONCLUSIONSJ wave is more likely seen in males and aged people and is more likely located in leads of inferior wall, than in leads of left and right walls.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Death, Sudden, Cardiac ; epidemiology ; prevention & control ; Electrocardiography ; Female ; Humans ; Male ; Middle Aged ; Physical Examination ; Prevalence ; Young Adult

Objective To explore the possible role of voltage-gated potassium channel-interacting protein 1 (KChIP1) in the pathogenesis of epilepsy. Methods Sprague Dawley female adult rats were treated with pentylenettrazole (PTZ) to develop acute and chronic epilepsy models. The approximate coronal sections of normal and epilepsy rat brain were processed for immunohistochemistry. Double-labeling confocal microscopy was used to determine the coexistence of KChIP1 and gamma-aminobutyric acid (GABA). Results KChIP1 was expressed abundantly throughout adult rat brain. KChIP1 is highly co-localize with GABA transmitter in hippocampus and cerebral cortex. In the acute PTZ-induced convulsive rats, the number of KChIP1-postive cells was significantly increased especially in the regions of CA1 and CA3 (P < 0.05); whereas the chronic PTZ-induced convulsive rats were found no changes. The number of GABA-labeled and co-labeled neurons in the hippocampus appeared to have no significant alteration responding to the epilepsy-genesis treatments. Conclusion KChIP1 might be involved in the PTZ-induced epileptogenesis process as a regulator to neuronal excitability through influencing the properties of potassium channels. KChIP1 is preferentially expressed in GABAergic neurons, but its changes did not couple with GABA in the epileptic models.