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Objective:To explore the effect of recombinant human syntaxin-4(STX4) on lipopolysaccharide(LPS)-induced injury in islet β-cells(INS-1).Methods:Pancreatic islet β-cells(INS-1) were divided into Control (blank control), LPS (LPS treatment), LPS+ NC (transfection of negative control vector, LPS treatment), and LPS+ STX4 (transfection of pcDNA-STX4, LPS treatment) groups. RT-qPCR and Western blot were used to detect STX4 mRNA and protein expression, flow cytometry to detect apoptosis, DCFHDA method to detect reactive oxygen species(ROS) level, xanthine oxidation method to detect superoxide orgotein dismutase(SOD) level, colorimetric method to detect glutathione peroxidase(GSH-Px) level, ammonium molybdate colorimetric method to detect catalase(CAT) level, thiobarbituric acid method to detect malonaldehyde(MDA) level, ELISA method to detect the level of interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and insulin secretion levels under glucose conditions secreted by cells, Western blot method to detect Cleared Caspase-3, Bcl-2 Associated X Protein(Bax), p65 protein expression. After treatment with NF-κB signaling pathway activator, STX4 up-regulated islet β-cell INS-1 was given LPS stimulation, and the same method was used to measure apoptosis, ROS, SOD, GSH-Px, CAT, MDA levels and secreted IL-1β, TNF-α, insulin levels and Cleaved Caspase-3, Bax, p65 protein levels.Results:Compared with the Control group, the expression of STX4 mRNA and protein in islet β cells of the LPS group decreased, the apoptosis rate, ROS level, and MDA levels increased, and the levels of SOD, GSH-Px, and CAT decreased, the levels of IL-1β, TNF-α increased, the level of insulin secreted by the cells decreased, and the expression levels of Cleaved Caspase-3, Bax, and p65 also increased. NF-κB pathway activator treatment reversed the effect of up-regulated STX4 on islet β-cell apoptosis, ROS, SOD, GSH-Px, CAT, MDA levels and secreted IL-1β, TNF-α levels, and Cleaved Caspase-3 , Bax and p65 protein levels.Conclusion:Up-regulation of STX4 alleviated LPS-induced islet β cell oxidative damage, apoptosis and inflammatory factor release. The underlying mechanism might be related to the inhibition of activated NF-κB signaling pathway.
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OBJECTIVE：To establish a method for the simultan eous determination of the contents of 12 flavonoids in Quzhiqiao. METHODS ：HPLC method was adopted. The determination was performed on Agilent Extend C 18 column with mobile phase consisted of 0.1％ formic acid-acetonitrile （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 35 ℃. The detection wavelength was set at 330 nm，and sample size was 10 μL. The contents of 12 components（such as eriocitrin，narirutin，naringin，naringenin，hesperidin，neohesperidin，hesperide hydrate ，luteolin，hesperide，nobiletin，hesperetin and hesperidolactone ）in 10 batches of Quzhiqiao from different collection places were determined. RESULTS ：The linear range of eriocitrin，narirutin，naringin，naringenin，hesperidin，neohesperidin，hesperide hydrate ，luteolin，hesperide，nobiletin，hesperetin and hesperidolactone were 1.65-16.51，4.50-45.02，35.41-354.12，4.11-41.12，2.29-22.86，34.96-349.56，1.42-14.15，1.50-15.04， 1.83-18.28，1.51-15.08，1.61-16.12，1.28-12.84 μg/mL，respectively（all r＞0.999 7）. The detection limits were 0.165 1，0.450 2， 3.541 2，0.411 2，0.228 6，3.495 6，0.141 5，0.150 4，0.182 8，0.150 8，0.161 2，0.128 4 μg/mL，respectively. The limits of quantitation were 0.547 8，1.487 4，11.663 3，1.360 3，0.758 3，11.594 9，0.466 3，0.497 1，0.601 2，0.499 9，0.532 3，0.424 6 μg/mL，respectively. RSDs of precision （n＝6），reproducibility（n＝6）and stability （24 h，n＝7）tests were all lower than 3％. The average recoveries were 99.50％，99.61％，98.18％，98.85％，98.48％，98.50％，98.25％，99.91％，103.13％，98.82％， 98.44％ ， 100.29％ （RSD＝1.49％ -2.38％ ， n＝6）. The contents of the above 12 components in 10 batches of samples from different collection places were 1.995 5-2.648 8，4.317 7- 5.005 1，33.215 5-34.054 6，3.140 4-3.471 5，3.221 2-3.748 8， 42.746 6-44.026 6，0.202 7-0.239 4，0.191 2-0.208 8，0.080 3- 0.097 9，0.291 9-0.307 1，0.119 9-0.149 1，0.082 7-0.089 8 mg/g. CONC LUSIONS：The method is accurate ，reliable，simple and efficient，which can be used to simultaneous determination of the contents of 12 flavonoids in Quzhiqiao ，and to provide reference for the establishment of quality control standards of Quzhiqiao.
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Objective:To evaluate the effects of dexamethasone on systemic lupus erythematosus complicated with cognitive dysfunction.Methods:Ten wild type mice and 20 MRL/lpr mice were applied for the research.MRL/lpr mice were randomly assigned to a MRL/lpr group and a MRL/lpr + dexamethasone (1.5 mg/kg) group.Interleukin-6 (IL-6),IL-1β,and tumor necrosis factor alpha (TNF-α) in serum and hippocampus were detected.The protein phosphorylation levels of phosphoinositide 3-kinase (P-PI3K),protein kinase B (P-Akt),NF-kappa-B inhibitor alpha (P-IκBa) and nuclear transcription factor kappa-B p65 (P-NF-κB p65) were detected by Western blot,the level of P-NF-κB p65 also was detected by immunohistochemistry.Results:Treatment with dexamethasone (1.5 mg/kg) alleviated the cognitive dysfunction and decreased the levels of IL-6,IL-1 β and TNF-α in serum and hippocampus,and reduced the levels of P-PI3K,P-Akt,P-IκBa and P-NF-κB p65 in hippocampus in MRL/lpr mice.Conclusion:Dexamethasone may play a protective role in the cognitive function by decreasing the levels of TNF-α and IL-1 β in the hippocampus of MRL/lpr lupus mice.
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Teaching competition is an effective way for college and university teachers to improve their teaching skills. Based on the teaching practice and experience in medical parasitology,this paper discusses several key issues in teaching competition including topics,teaching designs and teaching methods. It provides references for the teachers in department of parasitology of universities and colleges to improve the quality of classroom teaching.
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Objective To explore the application and significance of multiple clinical pathway training oriented teaching model in clinical teaching of laboratory diagnostic. Methods Totally 50 medical students enrolled in the Second Military Medical University from September to December in 2015 were divided into experimental group and control group. The course consists of theoretical teaching and experi-mental operation. The pathway group (n=25) were introduced into multiple clinical pathway training oriented teaching method. The theoretical teaching was carried out bysimulation examination application, simulation interpretation and simulation diagnosis and treatment, while the experimental course was carried out by using video teaching combined with actual operation. The control group was taught by traditional teaching method using slide teaching and operation display. The theoretical test including case study and operational skill tests were performed among students in both groups after 10 class hours training . The satisfaction questionnaires were conducted to evaluate the effectiveness and satisfaction of teaching guided by clinical pathway. Differences were compared with independent sample t testing using GraphPad Prism 5.0 statistical software. Results The medical records about professional theoretical test including case study and opera-tional skill test in the pathway group were superior to those in the control group with significant statistical difference (both P<0.05). The records of medical students were (81.84±7.21), (42.00±2.79) in the pathway group and (76.24 ±6.98), (37.00 ±3.71) in the control group. The questionnaire result showed that the pathway group's satisfaction was high, especially with the theoretical knowledge andsceneteaching (higher than 80％). The pathway group believed that multiple clinical pathway training helped to improve learning interest and clinical thinking ability . Conclusions Multiple clinical pathway training oriented teaching model is helpful for the medical students to achieve the basic idea of clinical pathway, improve the profes-sional ability, enhance the interest of learning and the quality of teaching, standardize teaching and promote teaching and learning.
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Objective To explore the anti-tumor effect of 17XL strains of Plasmodium yoelii(P.y)infection on melanoma in mice. Methods B16F10 tumor cells were axillarilly injected into the right flank of 20 C57BL/6 mice to establish tumor-bearing mouse models. The next day,the mice were randomly divided into a P.y infection group and control group,10 mice each group. Each mouse of the P.y infection group was intraperitoneally injected with 1×106 red blood cells including 20% P.y infection red blood cells,and each one of the control group were intraperitoneally injected with 1×106 normal red blood cells of C57BL/6 mice. The time of tumor formation of the mice in the two groups was observed and the tumor volumes were measured. Results The time of tumor formation in the P.y infection group[(11.30 ± 0.21)d]was significantly later than that in the control group [(10.40 ± 0.22)d](P < 0.05). From the tumors could be accurately measured to the study end point,both the tumors of mice in the two groups were growing,and the tumor volumes of mice in the P.y infection group were significantly less than those in the control group at each time point(all P < 0.05). The growth rate of tumors in the P.y infection group[(71.10 ± 6.29)mm3/d]was significantly slower than that in the control group[(302.80 ± 49.94)mm3/d](P < 0.05),and the growth rates of tumors every day in the P.y infection group were significantly slower than those in the control group(all P < 0.05). Conclusion The P.y in-fection can delay the occurrence of tumor and inhibit the growth of melanoma.
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Objective To understand the status of resistance to multiple insecticides and the frequencies of kdr mutations in Culex pipiens pallens from north?central Anhui Province. Methods From July to September,2014,the C. pipiens pallens mosquito larvae were collected in Huaibei,Bengbu and Chuzhou cities of the north?central Anhui Province and reared to adults. The female adult mosquitoes at 3-5 days post emergence were tested for susceptibility to the four insecticides,namely 0.05%deltamethrin,5%malathion,0.1%bendiocarb and 4%DDT,by using the standard WHO resistance tube bioassay. The detec?tion of the point mutations of the kdr gene at codon 1014 was conducted by PCR and DNA sequencing in the deltamethrin?resis?tant and?susceptible mosquitoes. Results High levels of resistance to all the four insecticides were found in all the three tested populations,although mosquito mortality varied among populations and test insecticides. Among the test insecticides,DDT showed lowest mortality with no significant difference(F=1.027,P>0.05)in all test populations,whereas significantly differ?ent mortalities were observed among populations for the remained three insecticides tested(deltamethrin,malathion,and ben?diocarb)(F = 23.823,33.955,128.841;all P < 0.01). Two types of non?synonymous kdr mutation at codon position 1014 (L1014F and L1014S)were observed. A positive correlation between L1014F mutation frequencies and deltamethrin resistance levels were detected in the three mosquito populations(r2=0.718,P<0.01). Conclusions The observed high levels of resis?tance to multiple?insecticides coupled with the occurrence of medium to high kdr frequencies in populations of C. pipiens pallens could profoundly affect the mosquito vector control programme in China. The local health departments need to strengthen vector dynamic monitoring and implement rational resistance management strategies.
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Objective To clone a gametocyte specific protein Pfgdv1 of Plasmodium falciparum,express and identify re?combinant Pfgdv1 protein in vitro. Methods PCR was performed to amplify Pfgdv1 from P. falciparum DNA which was got from the patient who was infected with P. falciparum,and the PCR product was inserted into pET28a(+)vector. pET28a?Pfg?dv1 recombinant plasmid was constructed and transformed into E. coli host BL21(DE3+). IPTG was used to induce the recombi?nant Pfgdv1 protein fused with His tag,and the protein was purified by His?NTA affinity chromatography. The recombinant pro?tein was identified by SDS?PAGE and Western blotting. Results The PCR product of Pfgdv1 gene was about 1.65 kb,meeting the expectation of predicted fragment size. The recombinant protein was about 67 kDa,which could be recognized by His?Tag monoclonal antibody. Conclusion The Pfgdv1 gene of P. falciparum is successfully cloned,and the recombinant Pfgdv1 pro?tein is expressed,thereby providing an opportunity for further study on transmission blocking vaccine.
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Objective To construct recombinant Mycobacterium smegmatis vaccine expressing Cysticercus cellulosae cC1 anti-gen. Methods The recombinant pET28a-cC1 plasmid was extracted and double digested by Xho I and BamH I restriction en-zymes,and shuttle plasmid pMV261 was extracted and double digested by Hind III and BamH I restriction enzymes. Both frag-ments were modified by Klenow fragment to form blunt end,then the large fragments of cC1 and pMV261 plasmid were purified and ligated by T4 ligase enzyme. The recombinant pMV261-cC1 plasmid was constructed and sequenced. Then the pMV261-cC1 plasmid was transformed into Mycobacterium smegmatis by the electrotransformation method. The recombinant cC1-Mycobacterium smegmatis was induced by heat and identified by the Western blotting method with the sera of cysticercosis patients. In addition, the growth states of the Mycobacterium smegmatis and the recombinant cC1-Mycobacterium smegmatis were compared and the growth curves were drawn. Results The restriction enzyme and sequencing results showed that the recombinant pMV261-cC1 plasmid was successfully constructed. After heat induction,a 40 kD band was showed by PAGE analysis of cC1-Mycobacterium smegmatis. The Western blotting results showed that the sera of cysticercosis patients could recognize the 40 kDa band,which sug-gested that cC1 protein was expressed in Mycobacterium smegmatis. Compared with the Mycobacterium smegmatis,the recombi-nant cC1-Mycobacterium smegmatis showed no significant difference in proliferation characteristics. Conclusion The recombi-nant cC1-Mycobacterium smegmatis vaccine has been successfully constructed.
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Objective To analyze the clinical application of HLA donor specific antibodies (DSAs) detected by Luminex single antigen beads,and to discuss the impact of early intervention on renal function.Method In 64 cases of living-relative renal transplantation,DSA was detected using a Luminex single antigen assay before and after transplantation.The positive recipients were given large doses of intravenous irnmunoglobulin (IVIG) and increased doses of mycophenolate mofetil (MMF).The relationship between DSA and renal function was analyzed.Result DSA was negative in all recipients before transplantation.Ten cases of DSA positive recipients were found in HLA mismatch after transplantation.After the intervention,two cases of DSA positive recipients became negative,immunofluorescence intensity was decreased by more than 50％ in 6 cases,and no significant reduction was found in the other two cases.Antibody-mediated rejection (AMR) occurred in two cases of intervention ineffective recipients after 3 to 6 months and the renal function was impaired.Conclusion Dynamic monitoring of DSA using Luminex single antigen beads may timely predict changes of renal function.Early application of large doses of IVIG and increasing doses of MMF can reduce the incidence of AMR.
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Malaria remains a serious public health problem,especially in developing countries. With the deepening of the un-derstanding of vivax malaria,Plasmodium vivax is also attracting more and more attention. An effective drug treatment is the foun-dation of controlling or even eliminating malaria. In recent years,more and more reports of chloroquine-resistance Plasmodium vivax have been reported. Plasmodium vivax chloroquine resistance has been a focus problem in vivax malaria prevention and treat-ment. In this paper,the research progress on distribution situation,detection methods and molecular markers of Plasmodium vivax chloroquine resistance is summarized.
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Objective :To properly understand the bacterial distribution and susceptibility to antibiotics in thesinus of nasal sinusitis. Method:The mucosal and secretione in sinus were taken from 135 patients who sufferedwith single nasal sinusitis, by functional endoscopic sinus surgery, were cultured for bacteria. Meanwhile the an-timicrobial susceptibility was determined for ordinary antibiotics. Result: The bacterial growth was present in88.15 % of cases ,mixed bacterial growth was present in 52.10 % of all bacterial ,anaerobe growth was present in39.06％00 of all bacteria． the susceptibility to penicillin was in 53.12%0 of all bacteria and cefaolin was in 87.50%.The susceptibility to metronidazole was in 94.67 % of all anaerobes. Conclusion:We think the method that peni-cillin in combination with streptomycin and metronidazole, sufameth oxazole united with metronidazole, cefaolinadded metronidazole were effective in nasal sinusitis treatment before the antimicrobial susceptive examinationhad been done.