Background Choroidal neovascularization (CNV) is one of the causes of blindness in multiple eye diseases.Researches showed that complement system participates in the pathogenesis of CNV.Objective This study was to construct the recombinant of complement factor B-small interference RNA (CFB-siRNA) expression vector and to observe its inhibitory effect on human umbilical vein endothelial cells (ECV-304).Methods CFB gene primers were designed based on human CFB gene,and an expression vector of CFB-siRNA was constructed by inserting CFB-siRNA into pRNAT-U6.1/Neo plasmid.Recombinant plasmids were confirmed by the digestion analysis of restriction endonuclease,and all inserted sequences were verified by DNA sequencing.The recombinant pRNAT-U6.1/CFB-siRNA plasmid and the blank plasmid were transfected into ECV-304 cells in the CFB-siRNA group and blank plasmid group by electroblot,respectively,and non-transfected cells served as the normal control group.The cells were observed under the fluorescence microscope 48 hours after transfection,and the transfective efficiency was calculated.The relative expression of CFB mRNA in the cells of different groups was detected by semi-quantitative reverse transcription PCR (RT-PCR).MTT was employed to calculated the growth inhibitory rates of the cells 24,48 and 72 hours after transfection.The percentages of the cells in different cell cycles were detected by flow cytometry.Results The sequence of the target vector was identical to the designed sequence.The green fluorescence protein (GFP) was seen in both the CFB-siRNA group and the blank plasmid group.The relative expression levels of CFB mRNA were 0.07 ±0.04,0.14 ±0.02 and 0.14 ±0.03 in the CFB-siRNA group,the blank plasmid group and the normal control group,respectively,a significant difference was obtained among the three groups (F=233.05,P =0.00);the expression level of CFB mRNA in the CFB-siRNA group was significantly declined in comparison with the blank plasmid group and the normal control group (both at P＜0.05).The growth inhibitory rates of the cells were (23.45 ±0.01) ％,(33.48 ±0.02) ％ and (45.49±0.01) ％ at 24,48 and 72 hours after transfection,respectively,a significant difference was obtained among the three groups (Fgroup =212.99,P =0.00);the growth inhibitory rates in CFB-siRNA group were significantly higher than that in the blank plasmid group and normal control group (all at P＜ 0.05).The percentages of G1 phase cells were (44.4 ±0.5) ％,(25.8 ±0.4) ％ and (27.9 ± 0.6) ％ in the CFB-siRNA group,the blank plasmid group and the normal control group respectively,a significant difference was obtained among the three groups (F=58.98,P=0.00).The percentages of G1 phase and G2 phase cells in the CFB-siRNA group were significantly higher than those in the blank plasmid group and the normal control group (all at P＜0.05).Conclusions Recombinant pRNAT-U6.1/CFB siRNA inhibits the proliferation of ECV-304 cells effectively by arresting the cells in G1 intermediate phase of the growth cycle.

OBJECTIVETo investigate the effect of chronic endurance exercise on microcirculatory reserve capacity of biceps brachii in Chinese rowers and provide a certain basis for the date standard foundation of monitoring of functional status and the foundation of database of reserve capacity of blood of Chinese rowers.

METHODSEmpty stomach in the morning, 77 rowers from different groups and 24 common health people were noninvasive tested by using PeriFlux System 5000, the test indexes include the microcirculatory reserve capacity and other related indexes of biceps brachii. The test sites of all athletes were the same space in biceps brachii of the right side of body, there was no space differences of all athletes . All athletes were tested in the relatively stable functional status, common people were healthy. The test value included basic values and heating values, put the before and after heating of microcirculatory blood perfusion (MBP) as the microcirculatory reserve capacity.

RESULTSHeavyweight female (198. 97 ± 98. 81) > heavyweight male (183. 45 ± 64. 31) > lightweight male (151. 01 ± 65. 96) > lightweight female(140.53 ± 43.22) > common male people(127.21 ± 56.38) > common female people(103.54 ± 33.41), the microcirculatory reserve capacity of each group athletes were higher than common people, except the comparison between lightweight female and common male people, and there was no significant difference among the different group athletes.

CONCLUSIONChronic endurance exercise can improve the microcirculatory reserve capacity of rowers, especially the heavyweight rowers; the normal value of microcirculatory reserve capacity of heavy weight rowers should be more than 160, and lightweight rowers should be more than 120. There was no significant difference among different sex athletes, if the value of microcirculatory reserve capacity is significant lower than normal, it shows that athletes are in the state of fatigue.

Arm ; Athletes ; Female ; Humans ; Male ; Microcirculation ; Muscle, Skeletal ; blood supply ; Sports

As a member of G protein coupled-receptors superfamily, free fatty acid receptor 1 (FFAR1), is also known as GPR40, has been shown to regulate numerous pathophysiological processes in a variety of tissues and organs. The activated FFAR1 has a variety of biological functions. For instance, it can not only regulate metabolism of fatty acids and glucose, but also play an important role in immune inflammatory response, it may be a potential drug target for the treatment of various chronic inflammatory diseases. In this review, we focus on the recent researches of FFAR1's action in the regulation of pathophysiological processes, its molecular mechanism and new agonists development. At the same time, this review will take the discovery of series FFAR1 agonists as examples, and display the applied prospects of FFAR1.

Objective To study the effect of clinical nursing pathways on the postoperative efficacy and quality of life of laparoscopic Miles surgery for rectal cancer.Methods 60 cases of lower rectal cancer patients received laparoscopic Miles surgery from Jan 2005 to Jan 2008 were given clinical nursing pathway and common nursing treatment.Clinical curative effect and quality of life were evaluated and compared.Results The satisfactory ratio,average hospitalization time and average expense of hospitalization were(95.0±0.4)%,(14.5 ±1.2)days and(28 156.0±572.5)yuan respectively,while they were(75.0±2.6)%,(16.5±1.3)days and(34 245.0±785.6)yuan in the control group.The differences between the two groups were statistically significant(P<0.05).The observation group were lower than control group in aspects of postoperative complications and time of colostomy accepting and nursing(P<0.05).The preoperative qualities of life had no difference between the two groups(P>0.05),but observation group had higher postoperative quality of life (P<0.05).Conclusions Clinical nursing pathway could significantly reduce hospitalization day and fees,postoperative complications improve the quality of life and satisfactory ratio for rectal cancer patients accepted laparoscopic Miles surgery.It may be the best nursing choice for laparoscopic Miles surgery.

OBJECTIVETo investigate the optimal glycated haemoglobin (HbA1c) cut off points and evaluate the impact of HbA1c on diabetes and pre-diabetes in middle-aged and elderly population.

METHODSSubjects were recruited from Shanghai Changfeng Study. A total of 1973 community-based participants (age ⋝45) without known diabetes underwent oral glucose tolerance test (OGTT) by using a 75-g oral glucose load and HbA1c was measured by using high performance liquid chromatography (HPLC). Subjects were classified as normal glucose tolerance (NGT), pre-diabetes(impaired glucose regulation, IGR) and new diagnosed diabetes (NDD) per 1999 WHO criteria. Two tests are compared with receiver operating characteristic curve (ROC).

RESULTSAmong 1973 subjects, 271 (13.7%) were diagnosed as NDD and 474 (24.0%) as IGR by using OGTT. HbA1c was 5.7%±0.7% in this population. Use of 6.5% as the HbA1C cutoff point has sensitivity of 38.7% and specificity of 98.5%. We recommend 6.0% as a better cutoff value for diagnosis of diabetes in this population (AUC 0.829, 95% CI 0.798-0.860, P<0.001) with its sensitivity and specificity as 66.1% and 86.8%. For IGR, the results showed low sensitivity (44.9%) and specificity (66.7%) with an AUC of 0.571 for HbA1c when 5.8% was used as the cutoff point. Participants detected with HbA1c⋝6.0% were associated with nearly the same metabolic characteristics, including body mass index (BMI), blood pressure, lipid profile and urine albumin-creatinine ratio (uACR) compared with diabetic subjects detected by OGTT.

CONCLUSIONThe optimum HbA1c cutoff point for diabetes in our study population was lower than ADA criteria, and HbA1c may not be used to identify IGR.

Aged ; China ; Diabetes Mellitus ; diagnosis ; metabolism ; Female ; Glycated Hemoglobin A ; metabolism ; Humans ; Male ; Middle Aged ; Prediabetic State ; diagnosis ; metabolism

OBJECTIVETo identify genes regulated by HBV preS1-transactivated protein 2 binding protein 1 (PS1TP2BP1).

METHODSPS1TP2BP1 gene was amplified by polymerase chain reaction (PCR) technique and cloned into the eukaryotic expression vector pcDNA 3.1/my-c-His A. The mRNAs isolated from HepG2 cells transfected recombinant eukaryotic expression vector pcDNA 3.1/myc-HisA-PS1TP2BP1 and pcDNA 3.1/myc-HisA empty vector were used to construct subtractive library. The differentially expressed genes were identified and analyzed.

RESULTS35 differentially expressed clones were obtained. Colony PCR identified 15 clones with 200-1000 bp inserts. Sequence analysis identified 15 differentially expressed genes.

CONCLUSIONThis study provides data for further characterize the function of PS1TP2BP1.

Amino Acid Sequence ; Base Sequence ; Carrier Proteins ; Cloning, Molecular ; Gene Library ; Genetic Vectors ; Hep G2 Cells ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B virus ; Humans ; Molecular Sequence Data ; Protein Precursors ; genetics ; Trans-Activators ; genetics

OBJECTIVETo study the protective effect of Sanhuangyinchi Fang drug serum (SF) against hydrogen peroxide-mediated DNA oxidative damage in LO2 cells.

METHODSThe LO2 cells were randomly divided into the control group, H(2)O(2) group, SF groups (5%, 10%, and 15%) and vitE group. The morphological features of the treated LO2 cells were observed under inverted microscope. The viability of the treated cells was assessed with CCK-8 method, and the activity of SOD, CAT and GSH-PX were detected biochemically. Reactive oxygen species (ROS) levels, the content of 8-OHdG, and DNA damage of the cells were evaluated by flow cytometry, ELISA, and Comet assay, respectively.

RESULTSCompared with H(2)O(2) group, the cells in SF groups (10% and 15%) and vitE group showed higher cell survival rate (P<0.05) and higher SOD, CAT, GSH-PX (P<0.05) and ROS scavenging activities (P<0.01) with markedly decreases the content of 8-OHdG (P<0.01) and reduced tailing ratio, tail length, tail moment and Olive tail moment (P<0.05).

CONCLUSIONSF drug serum, especially at the concentration of 15%, can protect LO2 cells from H(2)O(2)-mediated DNA oxidative damage.

Cell Line ; Comet Assay ; DNA Damage ; Deoxyguanosine ; analogs & derivatives ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Hydrogen Peroxide ; toxicity ; Oxidation-Reduction ; Oxidative Stress ; Protective Agents ; pharmacology ; Reactive Oxygen Species

OBJECTIVETo study the hepatoprotective effect of the extract of Terminala catappa leaves (TCE) and the possible mechanisms underlying its protection on acute liver injury induced by D-Galactosamine (D-GalN).

METHODIn vivo: D-GalN-induced liver injury model was used to evaluate the effect of TCE on the activities of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in mice. Structure of liver was observed and liver mitochondrial swelling was measured following D-GalN injection without or with TCE. In vitro: D-GalN-induced primary cultured hepatocytes injury model was used to value the effect of TCE on cultured hepatocytes. Cell viability was measured by means of MTT assay, and the AST and superoxide dismutase (SOD) activities in supernatant of cultured cells were investigated also.

RESULTIn acute hepatic injury test, with oral pretreatment of TCE, remarkable rises in serum AST and ALT activities (2.95 fold and 3.35 fold) induced by D-GalN were obviously reversed and significant morphological changes were remarkably lessened. In addition, the decrease in sensitivity of mitochondrial swelling to the exotic Ca2+ stimulation induced by D-GalN was also prevented by TCE. In primary cultured hepatocytes of mice, it was found that incubation with TCE could prevent the decrease in cell viability in a dose-dependent manner. It was also found that both the increase in AST level (1.9 fold) and the decrease in SOD activity (48.0%) in supernatant of primary cultured hepatocytes induced by D-GalN could be inhibited by pretreatment of TCE.

CONCLUSIONTCE has hepatoprotective activity and the mechanisms underlying its protective effect may be related to its antioxidant activity and protection on both hepatocytes and liver mitochondria.

Animals ; Cells, Cultured ; Chemical and Drug Induced Liver Injury ; blood ; etiology ; pathology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Galactosamine ; Liver ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred ICR ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Pregnancy ; Protective Agents ; pharmacology ; Terminalia ; chemistry

OBJECTIVETo investigate the volatile fatty acids in gingival crevicular fluid (GCF) and to analyze the relationship between the levels of the volatile fatty acids and chronic periodontitis.

METHODSGCF samples taken from 37 patients with chronic periodontitis and 16 volunteers with healthy periodontal status were analyzed by capillary electrophoresis.

RESULTSThe detection frequencies and concentrations of succinic acid, butyric acid and valeric acid were significantly higher in GCF of chronic periodontitis than in that of healthy group. The detection frequencies of propionic acid had no statistic difference between the two groups, but the concentrations of it was significantly higher in inflammation group. We also found that the concentrations of succinic acid, propionic acid and butyric acid were significantly lower in shallow pockets than that in deep pockets.

CONCLUSIONSThe volatile fatty acids, especially succinic acid, propionic acid, butyric acid and valeric acid were associated significantly with the severity and inflammation of periodontal disease. The levels of succinic acid, propionic acid and butyric acid in GCF were related to pocket depth.

Adult ; Aged ; Butyrates ; analysis ; Case-Control Studies ; Chronic Periodontitis ; metabolism ; Electrophoresis, Capillary ; Fatty Acids, Volatile ; analysis ; Female ; Gingival Crevicular Fluid ; metabolism ; Humans ; Male ; Middle Aged ; Periodontium ; metabolism ; Propionates ; analysis ; Succinic Acid ; analysis

OBJECTIVETo investigate the effect of parathyroid hormone related protein (PTHrP) on proliferation of human osteoblasts (MG-63) under the circumstance of tension force in vitro.

METHODSAn apparatus was designed and fabricated by which force was loaded onto the cultured cells in vitro. Reverse transcription-polymerase chain reaction (RT-PCR) was used for measuring the expression of PTHrP mRNA and c-fos mRNA. The effect of tension force and different PTHrP dose(0, 0.01, 0.1, 1 nmol/L) on the proliferation of human osteoblasts were examined using flow cytometry.

RESULTSVarious forces of the mechanical stretching exerted different influences on the intensities of the mRNA' expression. The strain of 12% induced the most remarkable mRNA' expression. The mitogenesis happened in the group with tension force (12%) combined with PTHrP was more active than that in the group with PTHrP or tension' force only. Tension force combined with PTHrP induced significantly more c-fos mRNA than that of tension force only.

CONCLUSIONThe mechanical stretching can inevitably influence the expression of PTHrP mRNA. The most active mitogenesis happened in the group with tension force combined with PTHrP. The effect may be related with the signaling pathways of c-fos.

Cell Line ; Cell Proliferation ; Cells, Cultured ; Humans ; Osteoblasts ; Parathyroid Hormone-Related Protein ; RNA, Messenger