Objective:To explore the influence of RNAi mediated Slug silencing on the growth and metastasis of colon cancer in nude mice.Methods: HCT116 colon cancer cells use for 24 five-week-old nude mice implanted subcutaneously , established colon cancer xenograft model in nude mice ,all divided into blank control group ,negative control group and the experimental group ,each group had eight nude mices.All group were injected with saline , negative plasmid and lentivirus vectors respectively.Tumor growth was observed and draw tumor curved growth ,changes in tumor growth and lymph node metastasis between the groups were observed ,Slug gene and protein expression were detected by immunohistochemistry ,qRT-PCR and Western blot analysis.Results: Slug gene shRNA intervention group compared with the control group and negative control group ,tumor grew slower ,tumor mass was significantly reduced (3.08±0.31 vs 7.37±1.18,7.46±1.16,P＜0.01),experimental group of lymph node-positive rate was 36.3%( 4 /11 ) ,compared to the negative control group 77.8% ( 14/18 ) and the control group was 68.4% ( 13/19 ) ( P＜0.01 ).Conclusion: Targeted Slug RNA interference can significantly inhibit the growth of colon cancer in nude mice ,lymph node metastasis and the expression of the gene protein in cancer tissue ,Slug may be a potential molecular target for colon cancer gene therapy.

Objective To discuss the significance of abnormal expression of Rb,p53 and PCNA in patients with bladder carcinoma and the correlation with the tumor grade,clinical stage and relapse.Meth- ods Immunohistochemical method was used to detect the Rb,p53 and PCNA gene expression in 60 cases with bladder carcinoma.Results The positive rates of p53,PCNA in 60 cases of bladder cancer is both 40.0 %.There were 27(45 %)cases lack of Rb expression.Multiple altered expression was found in 30 cases (50.0 %).The abnormal expression of Rb,p53 and PCNA was closely related to the tumor grade,clinical stage and relapse.Conclusion The detection of multiple gene is more valuable than that of single gene. Multiple altered expression of Rb,p53 and PCNA and their synergistic action might play an important role in the genesis and development of bladder cancer.

Objective The use of extracorporeal membrane oxygenation (ECMO) as a treatment for the failure of cardiopulmonary function after cardiac surgery is increasing and has been reported to be 3% to 5% in the cases with congenital heart disease. We reviewed our experience with ECMO in children who received heart surgery for congenital heart disease and complicated with severe heart failure postoperatively. Methods Eight patients received ECMO, seven was due to the failure to wean from bypass and one had fulminant myocarditis. Import membrane oxygenator,veno-arterial mode ECMO and right atriumascending aortic cannulation were used in 7 cases and peripheral cannulation via femoral veno-artery route was used in 1 case.Supportive intervention persisted from 65 to 498 hours, with flow rate maintained at 80 to 120 ml per minute per kilogram body weight. Results Five patients died, with a mortality of 62.5%, and 3 cases discharged, with a survival rate of 38%. Bleeding occurred in 5 cases, thrombosis occurred in 2 cases, hemolysis was identified in 1 case and DIC was observed in 1 case.One case had liver failure and 2 cases had malnutrition. Oxygenator plasma leakage occurred in 2 cases. Mean arterial blood pressure increased significantly after the establishment of ECMO as compared with that before the procedure [( 60.2 ± 7.8 )mmHg vs. (48. 1 ± 5.2 ) mmHg, P≤0.05]. The arterial concentration of lactate decreased significantly, from (5. 1 ± 0. 8 )mmol per liter before ECMO to ( 3.6 ±0. 5 )mmol per liter after ECMO, P ＜0.05. Conclusion For patients who survived the congenital heart surgery and no residual anatomic deformity, ECMO can be used as early as possible as a treatment for severe heart failure which resulted from coexistent of left and right ventricular and pulmonary insufficiency. An overall mortality may be decreased by ECMO technique as it plays a substitution role for gas exchange in the lung. As a result, the concentration of oxygen and the airway pressure used during ventilation, and the resultant lung injury can be reduced. Appropriate strategies involve transfusion of fresh platelet and packed red blood cells, replacement of frozen plasma and blood products, as well as rational use of vasoactive drugs and heparin, and maintaining a stable internal environment. Following strategies are also recommended: using continuous arterio-venous hemofiltration and durable heparin-coated membrne oxygenator, reducing hemorrhagic complications, monitoring pressure on both side of the film, identifying plasma leakage carefully and reducing the mechanical complications.

Objective This study was conducted to investigate the clinical outcomes and safety of percutaneous coronary intervention (PCI) to the single-opened vessel lesion among patients with severe left ventricular systolic dysfunction. Methods Twenty-seven patients with severe left ventricular systolic dysfunction (ejection fraction≤35%) undergoing PCI were included. All the patients received PCI only to the single-opened vessel lesion under the conditions of: (1) There were limitations to open chronic total occlusion (CTO);(2) Single-opened vessel lesion was not calcified and tortuous. Clinical outcomes, including success rate of PCI, changes of symptoms in-hospital, brain natriuretic peptide (BNP) and left ventricular ejection fraction (LVEF) pre-and one week post-PCI, the major adverse cardiac events (MACE, including death, myocardial infarction and target vessel revascularization) at 30-days after discharged were observed. Results The success rate of PCI was obtained in all 27 patients(100%), and all the patients received drug eluting stent implantation. The symptoms improvement occurred in all patients and the NYHA class improved from grade Ⅳto grade Ⅲin 22 patients(81.5%) in-hospital. Significant differences were noted in the mean BNP and LVEF between pre-PCI and one week post-PCI, BNP[(2699.6±1104.7) pg/ml vs. (737.0 ± 261.7) pg/ml, P<0.05],LVEF[(26.9±5.7)%vs. (36.0±3.41)%, P<0.05)]. No MACE happened in-hospital and at 30-days follow up. Conclusions PCI only to the single-opened vessel lesion among patients with severe left ventricular systolic dysfunction under the condition of limitations to open CTO is safe and can significantly improve clinical outcomes in-hospital and at 30-days follow up, but it must be emphasized that single-opened vessel lesion not with obvious calcification and tortuosity.

BACKGROUND: Many data demonstrate that the components of soybean can lower blood lipid,suppress the growth of cancer cells and exert weak estrogenic activities. However,little is known about the effect of isolated soybean protein on the lifespan of the organism.OBJECTIVE: To observe the effect of isolated soybean protein on the lifespan of drosophila melanogasters and investigate the mechanism of effective anti-aging and anti-oxidation action.DESIGN: A controlled trial based on drosophila melanogasters.SETTING: Department of nutrition and food hygiene in a university.MATERIALS: The experiment was conducted in the Department of Nutrition and Food Hygiene,Public Health College of Xinjiang Medical University from March to June 2002. A total of 400 drosophila melanogasters of American wild type with half for each gender were provided by the Department of Nutrition and Food Hygiene, Public Health College of Xinjiang Medical University.METHODS: The 400 drosophila melanogasters were divided into control group(normal culture) and three-dosage experiment groups(normal culture contained isolated soybean protein 0.2%,1.0% and 5.0%,respectively).From the second day on, the number of living and dead drosophila melanogasters was observed and counted until all died. Meanwhile, mean lifespan,half death time and maximal lifespan were calculated.MAIN OUTCOME MEASURFS: Mean lifespan, half death time and maximal lifespan of the drosophila melanogasters.RESULTS: Compared with that of control group, the lifespan of male and female drosophila melanogasters in experiment groups was prolonged by isolated soybean protein and responded in a dose-dependent manner,especially in high-dosage group. The mean lifespan, half death time and maximal lifespan of both female and male drosophila melanogasters were prolonged by 24.5% and20.7%,27.1% and22.0%,and 13.9% and 10.6%,respectively.CONCLUSION: Isolated soybean protein may have anti-aging and lifespan-prolonging effects on drosophila melanogasters.

The change of the effective components (liquiritin, glycyrrhizic acid, liquiritigenin, isoliquiritigenin) contents of Glycyrrhizae Radix et Rhizoma (GRR) before and after compatibilities in Sini decoction was studied in this paper. Taking single GRR decoction, GRR-Aconiti Lateralis Radix Praeparata (ALRP) decoction, GRR-Zingiberis Rhizoma (ZR) decoction and Sini decoction as test samples, the contents changing of the four effective components of GRR were measured by HPLC. The results showed that the contents of the four effective components of GRR in the single GRR decoction was higher than that in other samples, and the sequence was single GRR decoction > GRR-ZR decoction > GRR-ALRP decoction > Sini decoction. The contents of liquiritin were 11.18, 9.89, 9.67, 9.17 mg · g(-1); the contents of glycyrrhizic acid were 20.76, 15.58, 11.30, 8.52 mg · g(-1); the contents of liquiritigenin were 0.66, 0.57, 0.45, 0.24 mg · g(-1); the contents of isoliquiritigenin were 0.14, 0.07, 0.03, 0.01 mg · g(-1). Therefore, the effective components of GRR decreased obviously after GRR compatibility with ZR providing scientific basis for GRR relieving the strong nature of ZR. The effective components of GRR decreased sharply after GRR compatibility with ALRP providing scientific support for the material foundation research of GRR reducing the toxicity of ALRP. The effective components of GRR decreased further in Sini decoction indicating that the three medicines in Sini decoction were interactional, which reflecting the scientific connotation of the mutual-restraint/mutual-detoxication, mutual-promotion/mutual-assistance compatibilities in Sini decoction.
Chromatography, High Pressure Liquid ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; analysis ; Glycyrrhiza ; chemistry ; Rhizome ; chemistry

To clone cDNA of human leptin gene and obtain leptin protein for future study on leptin binding proteins. The cDNA of human leptin with 6 x his-tag was cloned by over-hang extension PCR protocol using human genomic DNA as template, and subcloned into in vitro expression vector pIVEX2.3MCS, and the fusion protein was expressed in vitro by Rapid Translation System (RTS) (RTS500 cycle primer Kit and RTS500 ProteoMaster of Roche company). The apparent molecular weight(19.46 kD) and the immuno-specificity of the fusion protein were confirmed by SDS-PAGE and Western blot, and the expressed fusion protein stayed mainly in the supernatant of the reaction mixture in soluble form. This work provides us solid basis for further study on new leptin-associated proteins.
Blotting, Western ; Cloning, Molecular ; DNA, Complementary ; genetics ; Electrophoresis, Polyacrylamide Gel ; Humans ; Leptin ; chemistry ; genetics ; metabolism ; Molecular Weight ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; chemistry ; genetics ; metabolism

OBJECTIVETo study on effects of acupuncture at a same point at physiological or pathological state on brain functional imaging.

METHODSElectroacupuncture was given at Neiguan (PC 6) of the healthy senile persons (normal group) and the patients of Alzheimer's disease (AD group), and they were scanned on a 1.5 T MRI scanner and the data of functional magnetic resonance imaging (fMRI) were analyzed by Statistical Parametric Mapping (SPM99). The functional imaging of different brain regions were obtained from both the healthy group and the AD group.

RESULTSBoth the frontal lobe and the temporal lobe were activated in varying degrees by electroacupuncture at Neiguan (PC 6) in the healthy group. However, the frontal lobe, the temporal lobe and the cingulate gyrus as well as the cerebellum were activated in varying degrees and other brain regions were not activated by electroacupuncture at Neiguan (PC 6) in the AD group.

CONCLUSIONAcupuncture at Neiguan (PC 6) can activate functions of different brain regions in the healthy group and the AD group, indicating that acupuncture at Neiguan (PC 6) has different effects on functional magnetic resonance imaging in different functional states.

Objective To evaluate the method of establishment of a minipig model of ischemic heart failure(HF) with acute myocardial infarction(AMI) by coronary balloon occlusion and coadministration of injecting of microthrombi and plastic microspheres.Methods A total of eighteen minipigs were selected.After coronary angiography,angioplasty balloons were placed in the mid-distal of left anterior descending(LAD).The balloon was inflated intermittently to occlude the LAD 3 times and then to occlude it continuously for 120 minutes.After the balloon was taken out,4F Judkins-type angiogrphic catheter was superelectively engaged in LAD and 3 mL intermixture of mierothrombi and plastic microspheres were injected at 10 minites interval until TIMI myocardial perfusion was grade<2 and left ventfieular end-diastolic pressure was maintained from 15 to 18 mmHg.Electrocardiogram(ECG),hemodynamic perameters,ultrasonic cardiogram,cTnI and CK-MB were measured.Myocardial infarction area was evaluated by histopathology.Results Fourteen days later,fifteen minipigs survived and fourteen satisfied the criteria(pulmonary capillary wedge pressure.PCWP>18 mmHg and eardio output (CO) decreased beyond 30% ). The changes of ECG, hemodynamic perameters, CKMB, cTnI and cardiac pathologic examination were in accordance with AMI. Conclusion A stable experimental method of establishment of minipig model of ischemic heart failure (HF) with acute myocardial infarction (AMI) by coronary balloon occlusion and coadministration of injecting of microthrombi and plastic mierospheres is succeded. This method has advantages such as closed chest, higher success rate and stability compared with the drug induced, taehycardia-pacing induced, coronary artery ligation induced or microsphere injection alone methods.

Objective To investigate the effects of HMGB1 small interference RNA (siRNA) on retinal vascular endothelial cells.Methods siRNA was used to inhibit the expression of HMGB1,followed by the application of CCK8 assay,Hochest33342 staining and flow cytometry to observe the effects of HMGB1 siRNA on retinal vascular endothelial cells in high glucose environment.Meanwhile,the expression of proteins related to apoptosis was detected by Western blot.Results The transfection of HMGB1 siRNA down-regulated the protein expression level of HMGB1 by 73％ in siRNA group compared with normal control (NC) group (P ＜ 0.05),and the protein expression level of HMGB1 in siRNA group was decreased by 75％ compared with scr-siRNA group (P ＜ 0.05).There was no significant difference between NC group and scrsiRNA group (P ＞ 0.05).The total apoptotic rate of NC group,high-glucose group,scrsiRNA group and siRNA group was (0.40 ± 0.03)％,(49.80 ± 3.50)％,(47.60 ±1.98) ％ and (23.60 ± 2.40) ％ by flow cytometry.Compared with NC group,the apoptotic rates of high-glucose group,scr-siRNA group and siRNA group were increased (all P ＜ 0.05).Compared with scr-siRNA group,the apoptotic rate of HRECs in siRNA group was reduced,with significant statistical difference (P ＜ 0.05).There was no significant difference in the rate of cell apoptosis between scr-siRNA group and high-glucose group (P ＞ 0.05).Compared with the NC group,the protein expression level of cleavedcaspase3 protein in high-glucose group and scr-siRNA group were increased by (233 ±10) ％ and (266 ± 22) ％,respectively (both P ＜ 0.05);compared with scr-siRNA group,the protein expression level of cleaved-caspase 3 in siRNA group was reduced by (43 ±3) ％ (P ＜ 0.05);and there was no significant difference in the protein expression of cleaved-caspase 3 in high-glucose group and scr-siRNA group (P ＞ 0.05).Compared with the NC group,the protein expression of B-cell lymphoma-2 (Bcl-2) in high-glucose group and scr-siRNA group was decreased by (32 ± 2) ％ and (29 ± 3) ％,accordingly (both P ＜ 0.05);compared with scr-siRNA group,the protein expression level of Bcl-2 in siRNA group was increased by (42 ± 2) ％ (P ＜ 0.05);and there was no significant difference in the protein expression of Bcl-2 in high-glucose group and scr-siRNA group (P ＞ 0.05).Conclusion HMGB1 siRNA can reduce the apoptosis of retinal vascular endothelial cells in high glucose environment by inhibiting the activation of cleavedcaspase3 and increasing the expression of Bcl-2.