Objective To explore the role of growth factor receptor-bound 2 (GRB2) in homo sapiens eukaryotic translation elongation factor 1 alpha 2 (EEF1A2) promoting the carcinogenesis and development of pancreatic cancer.Methods The human pancreatic cancer cell line SW1990 with low expression of EEF1A2,was transfected by Ad5/F35-EEF1A2 plasmid.The expression of EEF1A2 in human pancreatic cancer cell lines BxPC-3 cells with high expression of EEF1A2,was down-regulated by small interfering RNA (siRNA) interference.The expressions of GRB2 in SW1990 and BxPC-3 cells were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot before and after interference.The pancreatic cancer BxPC-3 cells were interfered by specific and efficient chemical synthesized siRNA fragment.The changes of BxPC-3 cell proliferation and migration were observed by methyl thiazoly tetrazolium (MTT) and Transwell assay before and after interference.The data were analyzed by one-way analysis of variance.Results After human pancreatic cancer cell line SW1990 was transfected with Ad5/F35-EEF1A2 plasmid,the expression of GRB2 at mRNA and protein level both increased significantly (F=5.67,6.39,both P＜0.05).After the expression of EEF1A2 was inhibited by EEF1A2-siRNA,the expression of GRB2 at mRNA and protein level both decreased significantly (F=6.59,4.69,both P＜0.05).After BxPC-3 cells was transfected with siRNA-GRB2 for 72 hours,the results of MTT assay indicated that the absorbance value was 1.22±0.14,compared with negative control group (1.42±0.15) and blank control group (1.39 ± 0.15) the difference was statistically significant (F =6.63,P＜ 0.05).The results of transwell assay showed that the migration ability of cells in siRNA-GRB2 group decreased.After 24 hours,the number of migrated cells was 24.10±4.25,compared with negative control group (54.72±5.24) and blank control group (51.26 ± 6.23) the difference was statistically significant (F=55.00,P＜ 0.05).Conclusion GRB2 is key molecule in EEF1A2 promoting the progenesis and development of pancreatic cancer.

The authors first describe the current situation of cooperative medical care in China, pointing out that due to low levels of guarantee, difficulties in fundraising and insufficient government reimbursement, cooperative medical care in rural areas have come across many hardships. It is, therefore, imperative to set up as soon as possible a health security system for people in rural areas. Firstly, it is necessary to break up the dual social security structure of urban and rural areas and set up a model with transitional goals based on the actual situation of the country. Secondly, it is suggested that security taxes begin to be collected, bringing the health security for people in rural areas into line with the social security framework so as to promote the health security for people in rural areas without lowering the level of social security in urban areas.

Objective To investigate the effects of over-expression of homo sapiens eukaryotic translation elongation factor 1 alpha 2 (EEF1A2) on in vitro invasion and lung metastasis of human pancreatic cancer SW1990 cells.Methods Letivirus-mediated delivery of EEF1A2 was used to enhance the expression of EEF1A2 gene in human pancreatic cancer SW1990 cells,the SW1990 cells stably over-expressing EEF1A2 protein (SW1990/EEF1A2 cells) were obtained,and the parent SW1990 cells and SW1990/GFP cells were used as the control,and the expressions of EEF1 A2 mRNA and protein were determined by Real-time PCR and Western blotting.The invasion ability of cells was determined by Transwell assay.The lung metastasis model was established by injection of SW1990 cells into the tail vein.Whole lung tissues were harvested,and visible nodules on tung surface were counted macroscopically 8 weeks later.Results The EEF1 A2 mRNA expression of SW1990/EEF1A2 was 3.252 ± 0.344,which was significantly higher than those in SW1990/GFP cells (1.000 ±0.060) and SW1990 cells (0.944 ±0.041,t =2.255,2.305,P＜0.01) ; the EEF1A2 protein expression was 0.833 ± 0.050,which was significantly higher than those in SW1990/GFP cells (0.247 ± 0.035) and SW1990 cells (0.273± 0.041,t=0.572,0.559,P＜0.01).The ability of invasion of SW1990/EEF1A2 cells was (60 ±4) cells,which was sigmificantly higher than (33 ±4) cells in SW1990/GFP group and (26 ± 3) cells in SW1990 group (t =31.33,34.78,P ＜ 0.01).Furthernore,SW1990/EEF1 A2 cells had a much higher incidence of lung metastasis in nude mice than SW1990/GFP cells and SW1990 cells in vivo (100％ vs.20％,20％,P ＜ 0.05).Conclusions EEF1 A2 over-expression can obviously increase the in vitro invasion and lung metastasis of pancreatic cancer SW1990 cells.

In this paper, Liuwei Dihuang pill was used to study the identification of Chinese patent medicine by fluorescence sequencing typing technology. The DNA of Paeonia suffruticosa was used as template to amplify by five pair of FAM fluorescence labeling primers. Then, the amplified products were sequenced. The sequencing results were analyzed by GeneMarker V1.80 to screen the best fluorescence labeling primers. As a result, psbA-trnH fluorescence labeling primer was used to identify the raw materials of Liuwei Dihuang pill. The results showed that three kinds of raw plant medicinal materials in Liuwei Dihuang pill were able to be correctly identified by psbA-trnH fluorescence labeling primer. The fluorescence sequencing typing technology can stably and accurately distinguish raw medicinal materials in Chinese patent medicine.
DNA Primers ; chemistry ; genetics ; DNA, Plant ; chemistry ; genetics ; Drugs, Chinese Herbal ; chemistry ; standards ; Fluorescent Dyes ; chemistry ; Plants, Medicinal ; chemistry ; genetics ; Polymerase Chain Reaction ; instrumentation ; methods ; Quality Control ; Staining and Labeling

Objective To investigate the factors associated with delay in decision to seek treatment in patients with acute myocardial infarction(AMI) in Beijing. Methods This prospective,cross-sectional,multicenter survey was conducted from November 1,2005 and December 31 ,2006. The participants consisted of 799 patients with STEMI admitted within 24 h of symptom onset to 19 hospitals in Beijing. Data were collected by semi-structured interviews and medical records review. The patients were categorized into an early decision group and the a late decision group based on the 30 min cut-off. Results The median(25%,75%) decision delay in STEMI patients was 60(20, 180)min. Factors associated with late decision in an univariate analysis were age ≥65 years, retirement or unemployment, history of myocardial infarction,symptom onset at home and intermittent symptoms, whereas presence of bystanders such as friends,coworkers or even strangers,unbearable symptoms,dyspnea,sweating,syncope and attribution of symptoms to cardiac origin were related to early decision. Multivariate logistic analysis showed that history of myocardial infarction,absence of syncope, intermittent symptoms,bearable symptoms and attribution of symptoms to noncardiac origin were independent predictors of decision delay＞30 min. Patients in the early decision group had more chances to receive acute reperfusion therapies(P=0.001) and shorter time intervals from symptom onset to reperfusion therapies(P＜0.001). Conclusions To a great extent patients with AMI in Beijing delayed in decision to seek treatment. History of myocardial infarction, symptom characteristics and symptom attribution were associated with decision delay.

OBJECTIVEScreening and characterizing high affinity completely humanized single-chain antibody (ScFv) against PreS1 of hepatitis B virus.

METHODSA combinatorial library of phage-displayed human ScFv, genes of which were derived from peripheral blood lymphocytes immunized by PreS1 of Hepatitis B Virus in vitro, was constructed. The library contained 7 10(8) clones.

RESULTSAfter 3 rounds panning, a high affinity (K=10(7) to 10(8) mol/L) ScFv specific to PreS1 was obtained. The V(H) belonged to human V(H4) family, and V(1) to V(4) by sequence analysis.

CONCLUSIONThis study suggests that the method of antigen stimulation in vitro is an expeditious way for the source of human immune antibody. And the ScFv may provide a more satisfactory therapy.

Antibodies, Monoclonal ; biosynthesis ; Antibody Affinity ; Hepatitis B Antibodies ; biosynthesis ; Hepatitis B Surface Antigens ; immunology ; Humans ; Peptide Library ; Protein Precursors ; immunology

Objective: To evaluate the effect of catheter ablation (CA) by endpoint of left atrium (AF)- pulmonary vein (PV) with bi-directional electrical isolation in treating the patients with paroxysmal atrial fibrillation (PAF) with remote electro-cardio graphic monitoring.
Methods: A total of 82 AF patients received radio frequency catheter ablation (RFCA) in our hospital and 76 PAF patients were observed. Based on circumferential pulmonary vein isolation (CPVI) judgment, the PAF patients were divided into 2 groups: Bi-directional block (BDB) group,n=20 and Entrance block (EB) group,n=56. The post-operative rhythm was followed-up by remote electro-cardio graphic monitoring and the clinical efifcacy of CA was prospectively observed.
Results:①All 76 patients finished CA, 1 patient died in EB group at 4 days after operation for pulmonary embolism, the rest 75 patients were followed-up for (31±19) months. The overall success rates of single ablation procedure at 3 months and 6 months after operation were 85.33% and 77.33%; in BDB group were 95.00% and 85.00%, in EB group were 81.82% and 74.55% respectively, the result was similar between 2 groups,P>0.05.②The overall occurrence rates of arrhythmia at 1 week and 3 months after operation were 35.53% and 17.33%; in BDB group were 15.00% and 5.00%, in EB group were 42.86% and 21.82% respectively, the differences between 2 groups were at P=0.049 andP>0.05.③Remote electro-cardio graphic monitoring recorded the patients with asymptomatic atrial arrhythmia at 1 week and 3 months after operation were at 22.22% and 23.08% respectively.
Conclusion:①Compared with EB group, BDB group had obviously lower occurrence rate of arrhythmia at short term after CA.②The overall success rate of single ablation procedure was similar between 2 groups which might be because less patients were studied.③Remote electro-cardio graphic monitoring has certain advantage for evaluating the occurrence of atrial arrhythmia after operation.

Objective To study the bystander effect of tumor necrosis factor related apoptosis inducing ligand(TRAIL) gene and it's mechanism. Methods Full length cDNA of human TRAIL was transfected into SMMC7721 with binary adenoviral vectors system. RT PCR was used to determine the expression of TRAIL gene. By MTT assay the effects of the TRAIL gene on the proliferation of SMMC7721 was studied; The apoptosis inducing ability of TRAIL gene on SMMC7721 was tested by fluorescence activated cell sorting (FACS). Bystander effect by testing the proliferation of the mixed cells of SMMC7721/TRAIL and SMMC7721 with different ratios, and the mechanism of bystander effect by testing the proliferation of SMMC7721 cultured with media of SMMC7721/TRAIL without cell components were also studied. Results TRAIL gene expressed by binary adenoviral vector system was able to inhibit proliferation(91.2%) and induce apoptosis(29.07%) of SMMC7721, significant difference between TRAIL gene and the other three controls were observed(PBS, LacZ, Bax)( P

AIM: To study the current density and function of Na + channel in cells from the epicardial border zone of the 1-week infarcted rabbit heart. METHODS: Rabbits were infarcted by ligation of the left anterior descending coronary artery. 1 week later, I Na was recorded using whole cell patch-clamp techniques in ventricular myocytes from infarcted heart(IZs) and compared with the I Na from noninfarcted heart(NZs). RESULTS: Peak I Na current density(at -30 mV) was significantly reduced in IZs(22 48?4 62 PA/PF, n= 14) compared with NZs(45 50?5 33 PA/PF, n= 12), P

AIM: To study the current density of transient outward potassium current (I_(to)) in cells from the epicardial zone of the 1-week and 2-month infarcted rabbit heart. METHODS: Rabbits were infarcted by ligation of the left anterior descending coronary artery, 1 week as well as 2 months later, the single ventricular myocytes were isolated enzymatically from the infracted area of 1-week infracted rabbit heart (PMI-1 week) and 2-month infracted heart (PMI-2 months), region remote from the infracted zone of 2-month infracted heart (REM-2 months) and free wall of left ventricule from noninfarcted heart (CON). I_(to) was recorded using whole cell patch-clamp techniques. (RESULTS:) Membrane capacitance of myocytes in REM-2 months group was signifitantly larger than that in CON. I_(to)current density (at +60 mV) was significantly reduced in PMI-1 week [(7.5?2.4) pA/pF, n=12] and PMI-2 months [(10.6?4.1) pA/pF, n=18] compared with CON [(17.4?5.2) pA/pF, n=16], P