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Background and purpose:MicroRNA(miRNA) is a class of small non-coding RNA playing an important regulatory role in many tumors. This study investigated which miRNA might negatively regulate the expression of Aurora-B in osteosarcoma cells, and to lay the foundation for the further investigation of the effort and regulation of Aurora-B in osteosarcoma malignant phenotype.Methods:Bioinformatics prediction software (http://www.targetscan.org) and luciferase assays were used to investigate which miRNA might target to modulate the Aurora-B. Real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot assay were used to further verify which miRNA could negative regulate the expression ofAurora-B gene.Results:Bioinformatics prediction showed let-7 family have the possibility to modulate the expression of Aurora-B; Luciferase assays showed thatAurora-B might be the target gene of let-7a/b/c/d/e/f/g/i; RTFQ-PCR and Western blot analysis testiifed that both the expression levels of Aurora-B mRNA and Aurora-B protein were signiifcantly decreased in Let-7a/g/i up-regulated U2-OS and HOS cells, compared to the cells in the negative control group; but in Let-7b/c/d/e/f up-regulated U2-OS and HOS cells, the expression levels of Aurora-B mRNA and Aurora-B protein have no signiifcant difference, compared to the cells in the negative control group.Conclusion:Let-7a/g/i may downregulate the expression of Aurora-B in human osteosarcoma cells.
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ABSTRACT:As the only intermediate host of Schistosoma j aponicum ,Oncomelania hupensis is an important link of schis‐tosomiasis .It plays an important role in the transmission of schistosomiasis .This article mainly demonstrates the following as‐pects :the invasion of schistosome miracidium into O .hupensis ,the growth of sporocyst ,and the mature and escape of cercari‐ae ,which would provide laboratory data from literatures for revealing the symbiotic relationship between O .hupensis and S . japonicum .However ,the symbiotic relationship between O .hupensis and S .japonicum is too complex to description com‐pletely .Therefore ,the symbiotic relationship will be the focus of future research .
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Objective To set up the molecular cytobiological model of endogenous coagulation factor Ⅷ (FⅧ) re-expressing in human liver cells L02,and to study the regulation pathway and molecular basis of the re-expression of FⅧ in L02 cells activated by NO signal.Methods The L02 cells at logarithm growth phase were selected and randomly divided into blank control group and experimental group, inhibitor group and inhibitor control group;they were cultured for 0,12,24,36,48,and 60 h.Flow cytometry was used to detect the expression of human FⅧ protein in L02 cells after treated for 48 h.Griess experiment was performed to detect the levels of NO in L02 cells at different time points;the transcription levels of human FⅧ gene,iNOS gene,NF-κB1 gene and I-κB alpha gene were detected by RT-PCR method.Western blotting method was used to detect the expression levels of human phosphorylated I-kappaB (phosphorylated I-κB)in L02 cells.Results The results of flow cytometry showed that the expression of human L02 FⅧ protein was found after treated with L-arginine for 48 h. The Griess results showed that the levels of NO in L02 cells in experimental group were significantly increased at 3,6,12,and 24 h (P<0.05)and the levels of NO in blank control group,inhibitor group and inhibitor control group had no changes. The RT-PCR results showed that the transcription of human FⅧ mRNA in L02 cells was found in experimental group,but there was no transcription of human FⅧ mRNA in blank control group,inhibitor group and inhibitor control group;the transcription levels of iNOS,NF-κB1 and I-κB alphain experiment group were increased(P<0.05)and the transcription levels of these genes in blank control group,inhibitor group and inhibitor control group had no changes. The Western blotting results showed that after adding L-arginine the expression level of phosphorylated I-κB was significantly increased (P < 0.05 ), other groups had no such change. Conclusion L-arginine can activate the phosphorylation of I-κB by NO signal pathway to lead to the changes in the expression of human FⅧ gene promoter upstream regulatory-related transcription factors NF-κB to activate the expression of human FⅧ in human liver cells L02.
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Objective High expression of the valosin-containing protein ( VCP) gene can enhance the metastasis of osteosar-coma via the AKT/PI3K/NF-KappaB/MMP-9 signaling pathway, but the molecular mechanisms underlying the up-regulation of VCP in osteosarcoma cells remains unknown .This study aimed to determine whether miRNA-129-5p can regulate the VCP expression and its targets in human osteosarcoma cells . Methods The microRNA target-predicting software TargetScanhuman 6.2 ( http://www.tar-getscan.org/) was used to predict the possible targets of miRNA-129-5p on the VCP gene.Then, two recombinant gene report vectors containing the wild VCP gene 3′UTR ( psi-VCP vector ) and mutant VCP gene 3′UTR ( psi-VCPmut vector ) were constructed , se-quenced, and identified.The human osteosarcoma U2-OS cells were co-transfected with miRNA-129-5p mimic and psi-VCP vector or psi-VCPmut vector, respectively.A non-specificity mimic transfection served as negative control , and the luciferase activity was detec-ted in each group. Results The software prediction showed only one conserved function site of miRNA-129-5p on the VCP gene 3′UTR163-169 bp.Luciferase activity was significantly lower in the psi-VCP vector +miRNA-129-5p transfection group (15.529 ± 1.902) than in the VCP control group (21.781 ±0.854), VCP mutation experimental group (19.978 ±1.377), and VCP mutation control group (21.952 ±1.516) (P<0.05), with no remarkable difference between the VCP mutation control and VCP control groups (P=0.276). Conclusion miRNA-129-5p can probably regulate the targets of the VCP gene in human osteosarcoma U 2-OS cells.
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Objective:To improve the sensitivity and the linear range of electrochemical immunosensor to detect Schistosoma japonicum (S.japonicum) antibody.Methods:Carbon inks and silver/silver chloride inks were printed on a polyethylene terephthalate (PET) board to make a two-electrode test strip,where carbon was the working electrode and S.japonicum soluble egg antigen (SEA) was fixed at one end of working electrode by different methods; silver/silver chloride electrode was used as control.We tested the valency of the antibody by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) in an electrochemistry workstation,and conducted comparison with the results of ELISA.Two new immunosensing electrodes have been developed,based on glutaraldehyde cross-linked (GA) or chitosan-glutaraldehyde cross-linked (Chit-GA) transducer fixing S.japonicum antigen.We tested the titer of the antibody by means of CV and DPV.Results:Our experimental S.japonicum antigen (50 μg/L) is the optimal test concentration for the GA sensor,and 10 μg/L for Chit-GA sensors.The immune reaction time of both electrodes is all essentially complete in 1 minute.The linear range for S.japonicura antibody in human positive serum sample detection by the glutaraldehyde cross-linked immunosensor is 1∶1000 to 1∶400,and by the chitosan-glutaraldehyde cross-linked immunosensor is 1∶1000 to 1∶500.As the concentration of dilution ratio of S.japonicum antibody in human positive serum sample increased,the test value of DPV increased proportionally.Conclusion:GA sensor and Chit-GA cross-linked S.japonicum sensors have high sensitivity and broad linear range response,and both exhibited a good linear relationship between the DPV signal and the test antibody titer.
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Objective To probe into the usage of statins in the patients with coronary artery disease in elderly patients. Methods From Jun 1,2009 to Mar 31,2009 in our hospital, 323 elderly outpatients with coronary artery disease were included in the investigation. The data were statistically analyzed. Results 62.2% of the patients used statins. The statins usage rates (44. 8% ,61.3% ,71.3% ) among three age groups(60 -69 years old,70- 79 years old, and ＞ 80 years old)were significantly different. The rates of statins usage were significantly different between male and female groups. Conclusion The usage of statins as secondary prevention of coronary artery disease varies and there is urgent need to standardize the usage of statins according to the treatment guideline.
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Objective To estimate heart function among patients with diabetic foot ulcer,and to investigate the characteristics of heart failure(HF) before and after treatment of ulcer. Methods Items associated with diabetes and some physiological and biochemical indicators were observed in patients with diabetic foot ulcer(162 cases) and with high risk factors of ulcer(75 cases). Heart function was evaluated at patients′ admission, during ulcer related treatment, and prior to discharge. Left ventricular ejection fraction and other cardiac assessment were measured with ultrasonic scan.Results During hospitalization, 23.6%(56/237) patients underwent HF with normal left ventricular ejection fraction, and it was 27.2%(44/162) in patients with foot ulcers. The prevalence of HF was 8.9%(21/237) in all patients studied on admission, and that was 10.5%(17/162) in patients with foot ulcer, more than that without foot ulcer(P＜0.01). More patients with HF were found, being 14.8%(35/237) during 2-7 days after ulcer related treatment initiated, and peak was on the 4th day. There was statistical difference among different Wagner grades(P＜0.05) about the morbidity rate of HF. All patients with HF were improved and tolerant to ulcer related treatment. Conclusion The prevalence of HF with normal left ventricular ejection fraction was relatively high in patients with diabetic foot ulcer, especially after ulcer treatment.
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Objective To prepare the infected Oncomelania hupensis by artificial method for the research on the activity, vaccine, and genetic variation of Schistosoma Japonicum (S. Japonicum).Methods The mature eggs of S. Japonicum were collected by Nylon silk method and the miracidia were incubated under appropriate conditions. Negative snails were infected with miracidia in different proportion by means of individual or collective infection to seek the best method and proportion of infection between miracidia and snails. Infected snails were divided into 12 groups in total. Ⅰ-Ⅵ groups were for individual infection and Ⅶ-Ⅻ groups were for collective infection. There were 200 snails in each group. The infection ratios between snails and miracidia in Group Ⅰ-Ⅵ or screened, numbered, and reared singly. The amount of cercariae was calculated once every 10 days until the infected snails died. Then cercariae shedding quantity, infection quantity, and mortality of infected snails in every group were compared to find the best infection method and the best infection proportion between miracidia and snails. The cercariae were collected from the first generation of infected snails and were used to infect experimental animals. The mature eggs of S. Japonicum were saved from the infected experimental animals and incubated to get miracidia. The snails were artificially infected by miracidium to get the second generation of infected snails. The developmental rates of adult worms, the egg density in fecal and liver were compared between artificially and naturally infected snails. Results In individual infection GroupⅠ-Ⅵ,the average infection value of snails were 0±0,22.7±4.2,31.7±4.5,53.0±5.3,39.3±5.9,32.7±4.7,the average fatality of snails were 21.7±3.1,25.0±3.6,31.3±4.9,44.7±6.5,78.3±9.5,89.7±13.6, and the average value of cercariae shedding from infected snails were 0.0±0.0,308.0±96.6,428.1±146.2,527.0±171.1,571.4±148.9,602.9±356.3, respectively. In collective infection Group Ⅶ-Ⅻ,the average infection value of snails were 0±0,12.3±2.5,18.7±4.7,28.3±4.2,33.3±4.7,29.3±5.5,and the average fatality of snails were 22.7±3.8,23.7±4.5,28.3±5.5,47.0±9.5,75.7±8.5,86.3±12.2, and the average value of cercariae shedding from infected snails were 0±0,244.5±57.3,292.3±74.8,347.1±100.8,477.2±142.1,447.3±161.4, respectively. The second generation of artificially infected snails was obtained successfully. The average infection rate and fatality rate for the second generation of artificially infected snails were 24.65% and 24.50%, both of which were not obviously different from that of the first generation of artificially infected snails (P>0.05). In the animal experiment, the worm growth rate for the naturally infected snails, the first or second generation of artificially infected snails were 68.50%,73.50% or 71.00%. There was no obvious difference among them (P>0.05). The fecal (or liver) eggs per gram for the naturally infected snails, the first or the second generation of artificially infected snails were 1 503±269,1 683±233, or 1 541±117 (or 6 641±1 819,6 272±1 419, or 7 263±1 643). There was no significant difference among the 3 groups (P>0.05). Conclusion Infected snails can be obtained through the artificial method by using S. Japonicum miracidia to infect snails. Individual infection has the advantage over collective infection. The optimal proportion of infection between first and the second generation of artificially infected snails in the average of cercariae shedding, infection, and fatality average of snails. There was no significant difference between artificially and naturally infected snails in the developmental rate of adult worms, fecal and liver eggs per gram.
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BACKGROUND: The Childhood Experience of Care and Abuse Questionnaire (CECA.Q) compiled by the Institute of Psychiatry, University of London, has been verified to have good reliability and validity in the assessment of foreign community adults and patients with affective disorder,but the application of CECA.Q in China still needs verification of reliability and validity in various population.OBJECTIVE: To analyze the reliability and validity of CECA.Q in community population.DESIGN: The subjects were randomized selected; the principle for the analysis of scale reliability and validity was followed.SETTING: Mental Health Institute of Central South University.PARTICIPANTS: From September 2003 to February 2004, 608 adults were selected from two communities of Hunan Changsha city and Henan Xinxiang city as the common community population group, totally 608scales of CECA.Q were sent out, 8 invalid ones were excluded, and finally 600 subjects finished the investigation. Sixty adult inpatients with depressive disease were randomly selected from Henan Provincial Psychiatric Hospital as the depression group.METHODS: The common community population and patients with depressive disease were investigated with CECA.Q, which was a self-rating scale mainly to retrospectively survey whether the adults had suffered from physical, sexual or mental abuse in their childhood, and the mental abuse had two factors of antipathy and neglect, and each included 8 items. The answers for the items were quantified into 5 grades. The subjects evaluated whether their parents had given any abuse to them; the higher the score was, the greater the mental abuse they suffered. Of the community population, 40 cases were the positive samples with the history of physical and sexual abuse in childhood evaluated by CECA.Q, and they were then assessed with Bifulco provided childhood experience of care and abuse (CECA) together with 40 cases randomly selected from 560 samples with negative evaluation. A total of 100 cases in the community population were also evaluated with Zung self-rating depression scale (SDS), besides, 30 cases were retested with CECA.Q at 2 weeks after the first evaluation.MAIN OUTCOME MEASURES: The internal consistency, test-retest reliability, contents and criterion validity of CECA.Q were analyzed. RESULTS: Of the community population, 8 invalid questionnaires were excluded, and the other 600 cases entered the statistical analysis. ① Testretest reliability of CECA.Q: The Cronbach α coefficient for the mental abuse section was 0.87 (0.85 for maternal one, 0.83 for paternal one); including that of antipathy factor was 0.84 (0.80 for maternal one, 0.81 for paternal one), and that of neglect factor was 0.85 (0.84 for maternal one,.83 for paternal one. The test-retest reliability coefficient for mental abuse as 0.83 (0.81 for maternal one, 082 for paternal one), including that of antipathy was 0.81 (0.82 for maternal one, 0.70 for paternal one), and that f neglect was 0.79 (0.78 for maternal one, 0.75 for paternal one). The twice evaluations of physical and sexual abuse had good consistency (Kappa=0.78). ② Criterion validity of CECA.Q: The correlation coefficients of mental abuse, antipathy and neglect factors in CECA.Q with Zung SDS were 0.58, 0.55 and 0.60 respectively (P＜0.01). CECA.Q and CECA had good consistency in evaluating history of physical and sexual abuse, and the score of mental abuse in the depressive patients was higher than that in community population (P＜0.05).CONCLUSION: The CECA.Q shows acceptable reliability and validity in community population, and can be used as a self-rating measure for childhood abuse experience in China.
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BACKGROUND: At present, the internationally public recognition of childhood trauma questionnaire is the authorized version of Bernstein,American psychologist, in 1998.OBJECTIVE: To set up the scale of Chinese version of childhood trauma questionnaire with 28 items and analyze its reliability and validity.DESIGN: Community investigation was designed.SETTING: Mental Health Institute, Xiangya Second Hospital, Central South University.PARTICIPANTS: Totally 441 students from 8 classes were randomized from a countryside middle school of a city in Henan in October 2004.METHODS: A total of 441 students were measured with childhood trauma questionnaire and 93 of them were re-measured 2 months later. Childhood trauma questionnaire included 28 items and divided into 5 subscales,named emotional abuse, physical abuse, sexual abuse, emotional neglect and physical neglect. Five grades were adopted in each item, named 1 score: never; 2 scores: occasionally; 3 scores: sometimes; 4 scores: often;and 5 scores: always. Every subscale was varied from 5 to 25 scores, and the total results were in the range from 25 to 125 scores. It was to analyze the internal identical property, reliability of re-measuring, average correlation coefficient among items and correlation coefficient between total score and every subscale. And the analysis was carried on validation factors. MAIN OUTCOME MEASURES: Homogeneity reliability, re-measuring reliability and validity, absolute fit index, relative fit index and parsimony index of childhood trauma questionnaire were involved.RESULTS: Totally 441 pieces of questionnaire were distributed on the spot and 435 pieces of questionnaire with integral and regular answers were collected. Totally 93 pieces of questionnaire were delivered for the second evaluation 2 months later and 93 pieces with regular answers were collected. All of those were used for the evaluation of re-measuring reliability of such measuring table. ① Cronbach α coefficient was 0.64 in Chinese version of childhood trauma questionnaire of and re-measuring reliability was 0.75. Cronbach αcoefficient of every subscale was varied from 0. 16 to 0.65 and re-measuring reliability was in the range from 0.27 to 0.73. The correlation coefficient among items was varied from -0.20 to 0. 44; ②The correlation coefficient between total score and every subscale was varied from 0. 36 to 0.68 and the correlation coefficient among subscales was from-0.01 to 0.39; ③ Indexes of validation factor analysis:The load coefficient of physical neglect was varied from 0.09 to 0. 64 and that in 2 items was less than 0.20. Multiple correlation coefficient was varied from-0. 20 to 0. 82, X2/df was 2.48 and root mean square error of appr0ximation(RMSEA) was 0.06. Added indexes: IFI (0. 76), CFI (0.75) and TLI (0.72) .Parsimony index: PNFI (0.58) and PCFI (0. 67).CONCLUSION: Chinese version of childhood trauma questionnaire provides better reliability and validity. According to validation analysis, except physical neglect, every index tallies with psychometric standards. The results of analysis on normative pathways are satisfactory, which explains that the subscales of such measuring model provide good matching property and conception validity.
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Percutaneous myocardial revascularization (PMR)，building on the succeeds of transmyocardial laser revascularization (TMR)，has developed that allows TMR-like channels to be cradated via a percutaneous approach.This catheter based approach avoids the morbidity associated with general anesthesia and thoractomy or thoracoscopy.It also avoids transmural treatment of the myocardium by the laser.Although its mechanism remains unclear,many patients with intractable angina and non-bypassable coronary artery disease have benefited from the new system.It is hoped that PMR will mimic the clinical benefit of TMR，and will gradually become a main method in treating end-staged coronary artery disease.