The rice gall dwarf disease, caused by the Rice gall dwarf virus (RGDV) is a serious disease occurring in rice in many regions of Guangdong province. As a basis to control the disease we have studied the genomic diversity of a variety of isolates from different locations. Genome segment 8(S8), encoding a main outer capsid protein (Pns8) of RGDV five isolates (BL, CH, DQ, GZ, XY) from Guangdong province was cloned and sequenced. The results revealed that all the S8 segments of the five isolates consisted of 1 578 nucleotides and had a single open reading frame (ORF) extending for 1 301 nucleotides from nucleotide 21 which encoded a polypeptide of 426 amino acids with an estimated molecular weight of 47.4 kDa. The S8 full-length sequence and the ORF sequence shared 97.3%-98.8% and 97.3%-99.1% nucleotide sequence identities within the five Chinese isolates, and shared 94.8%-95.6% and 95.0%-96.0% identities with those of the Thailand isolate respectively. The deduced amino acid sequence of Pns8 in GZ isolate was identical to that in the Thailand isolate, while the amino acid sequence variability of Pns8 within five Chinese isolates ranged from 0.5% to 2.1%. These results indicate that the S8 segment of RGDV is highly conserved in different isolates from different locations. The S8 cDNA from the XY isolate was cloned into the plasmid vector pET-28b(+) and a fused expression protein with an apparent molecular mass of 51kDa was specifically detected in an analysis of Escherichia coli Rossetta(DE3)Ⅱcells. To our knowledge, this is the first report on analysis of the RGDV segment 8 sequence and genetic comparison of different RGDV isolates and their protein expression.

Objective To investigate the effects of Tanshinone Ⅱ A (extracted from Chinese herb medicine Salvia miltiorrhiza Bge.) on ventricular arrhythmias of rabbit hearts induced by ischemia in order to illuminate its mechanism of anti-arrhythmia.Methods Thirty rabbits were randomly (random number)divided into normal group,ischemic group and Tanshinone Ⅱ A group.Model of wedge shaped mass of rabbit left ventricular myocardium with coronary perfusion was prepared,and then by using floating glassy microelectrode,the trans-mural ECG,QT interval,the trans-mural dispersion of re-polarization (TDR) and trans-membrane action potentials from both endocardium and epicardium were simultaneously and wholly recorded.The incidence of ventricular arrhythmia in myocardium was observed after ischemia for thirty min.Results Under the condition of acute ischemia,compared with normal group,the incidence of ventricular arrhythmia and TDR were significantly increased in ischemia group (P ＜ 0.01),while incidence of ventricular arrhythmia and TDR were significantly reduced in tanshinone ⅡA group compared with ischemia group (P ＜ 0.05).The incidences of ventricular arrhythmia in normal,ischemia and Tanshinone Ⅱ A groups were 0/10,9/10 and 2/10 respectively.Conclusions Tanshinone Ⅱ A prevents ventricular arrhythmia and reduces TDR significantly in ischemic rabbit hearts.

Objective To investigate the relationship between CYP2J2*7 mutation(G-76T) and coronary heart disease (CHD) in Chinese Hanpopulation and to study the effects of CYP2J2 geneover-expressionon the proliferation and migrationof aortic smooth muscle cells of ApoE-/- mice. Methods CYP2J2*7 genotype was detectedin 500 patients with CHD and 478 controlsubjects by the Polymerase Chain Reaction-Restriction Frag-ment Length Polymorphism (PCR-RFLP). Culturedaortic smooth muscle cells of ApoE-/- mice were divided into control group, sham transfectiongroup and CYP2J2 over-expression group. Cell proliferation and migration were investigated after CYP2J2 over-expressionby MTS and Transwell assay. Results The frequency of CYP2J2*7 in CHD group was significantly higher than that incontrol group (10.00% vs. 6.49%, P = 0.046). Same is the case in female cases(P = 0.026). Compared with these of aortic smooth muscle cells incontrol group and sham trans-fectiongroup, the cell proliferation in 24, 48, 72 h, and the cell migration in 48 h after CYP2J2 over-expression in CYP2J2 group were significantly suppressed. Conclusions CYP2J2*7 mutation might increase the risk of CHD in Chinese Han population. CYP2J2 over-expression can suppress the proliferation and migration of aortic smooth muscle cells and CYP2J2 might have the effect of anti-atherosclerosis.

The study was aimed to reveal the effects of matrix metalloproteinase-2 (MMP-2) on cell proliferation, migration, invasion, angiogenesis and cell cycle. The small interfering RNA (siRNA) of MMP-2 transfected into endothelial cells EAhy926, the transformation efficiency at protein and gene levels was evaluated by using flow cytometry and RT-PCR respectively. MTT method was used to detect the proliferation ability of EAhy926. The migration and invasion abilities of EAhy926 induced by two kinds of factors, type IV collagen (COL IV) and fibronectin (Fn) were assayed with Rose Bengal dying method. The changes of angiogenesis were determined by three-dimension culture. The changes of cell cycle and related gene expression were assayed by using flow cytometry and RT-PCR respectively. The results indicated that the proliferation ability of EAhy926 had no obvious difference between interference or not. The migration ability of EAhy926 induced by two kinds of factors, type IV collagen (COL IV) and fibronectin (Fn) was inhibited after interfered by siRNA for 48 hours, and was stronger inhibition to COL IV than Fn (Fn control: 0.581+/-0.012 vs 0.261+/-0.002; COL IV control: 0.467+/-0.009 vs 0.110+/-0.010, p<0.01). The invasion test had the similar result as migration test. (Fn vs control: 0.365+/-0.012 vs 0.101+/-0.002; COL IV vs control: 0.317+/-0.009 vs 0.102+/-0.010, p<0.01). The angiogenesis ability of endothelial cells dropped to 58.9% of control after interference with siRNA for 48 hours. In the cell cycle experiments, after RNAi for 48 hours and 72 hours, the cell ratio of G1 rose from [(65.9+/-2.53)%; (63.2+/-1.89)%] to [(83.9+/-2.53)%, (89.2+/-1.24)%]% (p<0.01); the cell ratio of S and G2 dropped from [(32.7+/-1.91)%, (37.1+/-2.65)%] to [(18.1+/-1.49)%, (10.2+/-0.85)%] (p<0.01). The results analyzed by semi-quantitative RT-PCR showed that the expression levels of Rb, cyclin D1 PCNA genes dropped to 35%, 51% and 22% of normal control respectively. It is concluded that the expression of MMP-2 does not distinctly correlate with the proliferation of endothelial cells, but plays a very important role in affecting endothelial cell migration, invasion and angiogenesis, and participates in regulating cell cycle through Rb, cyclinD1 and PCNA.
Angiogenesis Inhibitors ; Cell Line ; Cell Movement ; Cell Proliferation ; Endothelial Cells ; cytology ; Humans ; Matrix Metalloproteinase 2 ; genetics ; physiology ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo report the newly developed reconstruction technique after laparoscopic total gastrectomy: intracorporeal circular stapling esophagojejunostomy using the transorally inserted anvil (OrVil; Covidien), and evaluate its feasibility, safety, and clinical outcomes.

METHODSAfter LTG (3 patients with gastric carcinoma in the body) or LPG (2 patients with gastric carcinoma in the cardiac and fundus, respectively, and 1 with cardiac stromal tumor), the anvil was then inserted transorally into the esophagus by using the OrVil system. Double-stapling esophagojejunostomy or esophagogastrostomy with a circular stapler was performed intracorporeally.

RESULTSThe operations were uneventful. The operative time was (183.3+/-25.8) min, and blood loss was (128.3+/-90.2) ml. Postoperative fluorography revealed no anastomotic leakage or stenosis. Patients resumed an oral liquid diet on postoperative day (4.0+/-1.1), and were discharged on day (9.0+/-2.6). Patients were followed at 28 days and no complications were reported.

CONCLUSIONSLTG with Roux-en-Y reconstruction or LPG with esophagogastrostomy using the OrVil system appear to be safe and reliable with satisfactory short-term outcomes.

Anastomosis, Surgical ; Esophagus ; surgery ; Gastrectomy ; methods ; Gastric Stump ; surgery ; Humans ; Jejunum ; surgery ; Laparoscopy

Objective To investigate the effect of atorvastatin on oxidative stress and intracellular lipid accumulation in HepG2 cells under inflammatory state and explore the underlying mechanism.Methods HepG2 cells were treated with 100 ng · mL-1 TNF-α,100 ng · mL-1 TNF-α ± 10 μmol · L-1 atorvastatin in the presence of LDL for 24 h.Oil red O staining was used to examine the intracellular lipid contents.The mRNA and protein expressions of lipogenic genes (FAS,ACC and SREBP1) were detected by real-time polymerase chain reaction and Western blot.ROS levels were measured with the fluorescent probe of DCFH-DA.Contents of H2O2 and MDA were determined using the colorimetric method.Results Compared with normal group(the gray value of SREBP1 was 1.01 ± 0.001),the gray value of SREBP1 in model group was 1.61 ± 0.34.The mRNA levels in normal group of SREBP1,FAS,ACC respectively were 1.01 ± 0.16,1.03 ± 0.32,0.95 ± 0.29,the values in model group respectively were 3.61 ± 0.39,1.99 ± 0.36,2.37 ± 0.52,the differences were statistically significantly (P ＜ 0.05).Compared with model group,the mRNA levels of SREBP1,FAS,ACC and the gray value of SREBP1 in experimental group respectively were 2.95 ± 0.92,3.99 ± 1.16,2.85 ± 0.91,2.94 ± 0.65,the differences were statistically significantly(P ＜0.05).At the same time,compared with normal group,the levels of ROS(fluorescenceintensity),H2O2,MDA respectively were 1.00 ±0.20,and (2.30 ±0.31) (0.78 ±0.22) nmol · mg-1,the levels in model group respectively were 1.77 ± 0.25 and (4.32 ± 0.77),(1.86 ± 0.23) nmol · mg-1,the differences were statistically significantly (P ＜ 0.05).Compared with model group,the levels of ROS,H2 O2,MDA in HepG2 cells in experimental group respectively were 3.2 ±0.53 and (5.31 ±0.75),(3.43 ± 1.15) nmol · mg-1,the differences were statistically significantly(P ＜ 0.05).Conclusion Atorvastatin induced intracellular lipid accumulation in HepG2 cells under inflammatory stress,which may be associated with the increased oxidative stress.

Objective:To evaluate the gene mutation profile and prognostic significance of adult cytogenetically normal acute myeloid leukemia (CN-AML) with CEBPA-bZIP mutation. Methods:Targeted sequencing was implemented on the diagnostic bone marrow DNA samples of 141 adult CN-AML subjects with CEBPA-bZIP mutation. The nomogram model for leukemia-free survival (LFS) rate was generated by combining genetic abnormalities and clinical data. Risk stratification was conducted based on prognostic variables and the effect of risk-adjusted consolidation therapy was investigated by Kaplan-Meier method. Results:Four variables were finally included in our nomogram model after multivariate Cox analysis,and an equation for risk score calculation was obtained,risk score=1.3002×white blood cell (WBC) (≥18.77×109/L)+1.4065×CSF3R mutation positive+2.6489×KMT2A mutation positive+1.0128×DNA methylation-related genes mutation positive. According to the nomogram model,patients were further divided into low-risk group (score=0,n=46) and high-risk group (score>0,n=95). Prognostic analysis showed that the 5-year LFS rate,5-year overall survival (OS) rate,and 5-year cumulative incidence of relapse (CIR) of patients who received allogeneic hematopoietic stem cell transplantation (allo-HSCT) in the high-risk group were 93.5％,97.1％,and 3.5％,while those in patients who received maintenance chemotherapy were 32.9％,70.5％,and 63.4％,respectively. The differences were statistically significant (all P<0.05). Allo-HSCT could significantly improve the prognosis of patients in high-risk group. However,no corresponding benefit was observed in the low-risk group. Conclusion:Adult CN-AML with CEBPA-bZIP mutation has a complex co-mutation pattern. The nomogram model based on mutations of CFS3R,KMT2A and DNA methylation-related genes together with WBC count can further divide this subset of patients into a relatively low-risk group and a relatively high-risk group. For individuals in the high-risk group,allo-HSCT is proposed as post-remission therapy. The above data will benefit the prognosis estimation and treatment decision for adult CN-AML with CEBPA-bZIP mutation.

BACKGROUNDvon Willebrand factor (vWF) mediates the initial capture of platelets to vascular subendothelium and is essential for platelet aggregation under high fluid shear stress as in arterial stenosis. On release from endothelial cells, vWF is rapidly cleaved by ADAMTS13/vWF-cleaving protease (vWF-CP). We investigated the clinical significance of changes in plasma vWF and vWF-CP activities in chronic renal disease.

METHODSPlasma vWF and vWF-CP activities were measured using enzyme-linked immunosorbent assay (ELISA) and residual collagen binding assay respectively in patients with lupus nephritis (n = 31), primary nephritic syndrome (n = 25), diabetic nephropathy (n = 45), chronic glomerulonephritis (n = 38) and 40 normal controls. The relation of their levels with pathological and renal status was analyzed.

RESULTSIn all diseased patients the levels of vWF were significantly higher and vWF-CP activity significantly lower than the controls (both P < 0.01). vWF in the four subgroups did not correlate with the stage of disease but correlated negatively with vWF-CP activity. vWF-CP activity was not changed two weeks after renal transplantation. Renal biopsy demonstrated that the vWF level in stage IV was higher than in stages II and III while vWF-CP activity was lower in patients with lupus nephritis. After eight-week treatment, the vWF level significantly decreased and the vWF-CP activity significantly increased in systemic lupus erythema, disease activity index < 9, but not with index = 9. Even though the vWF-CP activity was significantly lower in membranous nephropathy than in minimal change disease, mesangial proliferative glomerulonephritis or IgA glomerulonephritis, the vWF level was not significantly different.

CONCLUSIONSThe alterations of plasma vWF and vWF-CP activities were associated with different renal pathologies. Injury to endothelial cells and autoantibodies against vWF-CP activity may result in higher vWF level and lower vWF-CP activity in chronic renal disease and thus a mechanism for worsening of chronic renal disease and thrombosis.

ADAM Proteins ; blood ; ADAMTS13 Protein ; Adolescent ; Adult ; Aged ; Chronic Disease ; Female ; Humans ; Kidney Diseases ; blood ; Kidney Transplantation ; Lupus Nephritis ; blood ; Male ; Middle Aged ; von Willebrand Factor ; analysis

OBJECTIVEThe development of pulmonary vascular bed is strongly flow-dependent. Abnormal pulmonary blood flow leads to pulmonary pathological changes. This study aimed to observe the pathological changes of small pulmonary arteries and alveoli in complex congenital heart defect with diminished pulmonary blood flow but without aortopulmonary collateral artery (APCA) and patent ductus arteriosus (PDA) in infants and young children.

METHODSAutopsy pulmonary specimens obtained from 5 children who died of non-cardiovascular diseases were used as the control group (age: 4-18 months). Fifty-six children (age: 4-36 months) with complex congenital heart defect with diminished pulmonary blood flow but without APCA and PDA served as the study group, including 34 cases of tetralogy of Fallot, 7 cases of double outlet right ventricle with pulmonary stenosis, 9 cases of single ventricle with pulmonary stenosis, 4 cases of tricuspid atresia with pulmonary stenosis and 2 cases of complete atrioventricular septal defect with pulmonary stenosis. Pulmonary specimen sections were stained by hematoxylin-eosin and Weigert-Van Gieson. Percentage of media thickness (MT%), percentage of media section area (MS%), number of small arterial per square centimeter (APSC), mean alveolar number (MAN), mean linear intercept (MLI), proportion of parenchyma area in total area (PPA%) and alveolar to small arterial ratio per unit area (AAR) were measured by morphologic quantitative analysis.

RESULTSMT% (10.93+/-2.87% vs 15.08+/-2.51%), MS% (18.97+/-5.56% vs 25.04+/-3.87%) and APSC (202.43+/-67.45 vs 441.69+/-65.29) decreased significantly in the study group compared with the control group (P<0.01). The internal diameter of small pulmonary artery (80.26+/-21.57 microm vs 58.53+/-10.29 microm; P<0.05), AAR (46.59+/-14.43 vs 34.46+/-4.98; P<0.01) and MLI (144.98+/-44.87 microm vs 108.39+/-20.76 microm; P<0.05) increased significantly compared with the control group.

CONCLUSIONSThe media of small pulmonary arteries becomes thinner, the lumen of small pulmonary arteries becomes larger, and the number of small arterial per square centimeter and the mean alveolar number are reduced in infants and young children with complex congenital heart defect with diminished pulmonary blood flow but without APCA and PDA.

Aorta ; abnormalities ; Child, Preschool ; Collateral Circulation ; Ductus Arteriosus, Patent ; pathology ; Female ; Heart Defects, Congenital ; pathology ; Humans ; Infant ; Lung ; pathology ; Male ; Pulmonary Artery ; abnormalities ; Pulmonary Circulation

OBJECTIVETo investigate the effects of single or combined administration of cedilanid and small-dose of enalaprilat on heart, liver, kidney and intestine damages at early stage of severe scald in rats.

METHODSForty healthy male Wistar rats were enrolled in the study and randomly divided into: sham, burn control, cedilanid, enalaprilat, cedilanid + enalaprilat groups, with 8 rats in each group. Rats, except that of sham group (simulated scald with 37 degrees C water) were inflicted with 30% TBSA full-thickness scald, and were injected with Ringer's lactate solution (4 mLxkg(-1)x1% TBSA(-1)) intraperitoneally 30 minutes after burn. Then rats in cedilanid group were given cedilanid injection (0.2 mg/kg) intravenously, and those in enalaprilat group were given enalaprilat (1 mg/kg), and cedilanid + enalaprilat group with cedilanid and enalapril in the same dosage. At 6 post burn hour (PBH) or sham injury, parameters of myocardiac mechanics were recorded with the Multiple Channel Physiological Signal Collecting and Processing System. The blood flow of the liver, kidney and intestine was respectively detected with the Laser Doppler Flowmetry at 6 PBH. Serum contents of cTnI, TBA, beta2-MG and DAO were determined at 6 PBH to reflect visceral damages.

RESULTSCompared with those in sham group, the parameters of myocardiac mechanics and blood flow of liver, kidney, intestine (158 +/- 32, 156 +/- 46, 119 +/- 30 PU, respectively) in burn control group were obviously decreased (P < 0.05), and the serum contents of cTnI, TBA, beta2-MG, DAO (5.0 +/- 0.3 microg/L, 82 +/- 23 micromol/L, 2.55 +/- 0.15 mg/L, 1.52 +/- 0.08 kU/L, respectively) in burn control group were obviously increased (P < 0.05). Compared with those in burn control group, the parameters of myocardiac mechanics and blood flow of liver, kidney, intestine in the cedilanid or enalaprilat groups increased markedly, and their serum contents of cTnI, TBA, beta2-MG, DAO decreased significantly (P < 0.05). Compared with those in burn control group, the parameters of myocardiac mechanics and blood flow of liver, kidney, intestine (240 +/- 49, 239 +/- 75, 194 +/- 55 PU, respectively) in cedilanid + enalaprilat group increased significantly (P < 0.05), and the serum contents of cTnI, TBA, beta2-MG, DAO (3.43 +/- 0.21 microg/L, 47 +/- 8 micromol/L, 2.01 +/- 0.16 mg/L, 1.17 +/- 0.15 kU/L, respectively) were decreased (P < 0.05).

CONCLUSIONSingle administration of cedilanid or small-dose enalaprilat can ameliorate impairment of cardiac functions, prevent damages to liver, kidney and intestine in early stage of severe scald in rats. Combined administration of cedilanid and small-dose enalaprilat seems to be more effective.

Animals ; Burns ; drug therapy ; pathology ; Deslanoside ; administration & dosage ; Disease Models, Animal ; Drug Therapy, Combination ; Enalaprilat ; administration & dosage ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Viscera ; pathology