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Objective@#To elucidate the association between large conductance calcium-activated potassium channels (BKCa) in the paraventricular hypothalamic nucleus (PVN) and sympathetic outflow in rats with chronic heart failure (CHF) .@*Methods@#Male Wistar rats (6-7 weeks old) were randomized to sham operated group and CHF group (coronary artery ligation) . Two weeks after operation, BKCa inhibitor Iberiotoxin (IBTX) was infused into PVN by osmotic minipumps, rats were divided into following groups: sham+aCSF, CHF+aCSF, sham+low dose IBTX (0.125 nmol/nl) , CHF+low dose IBTX, sham+moderate dose IBTX (1.25 nmol/nl) , CHF+moderate dose IBTX, sham+ high dose IBTX (12.5 nmol/nl) , and CHF+high dose IBTX (n=6 each) . Additional rats were grouped as follows: sham+vehicle, sham+KCNMB4 knockdown (by rAAV2-KCNMB4 shRNA virus injection in PVN) , CHF+vehicle, CHF+ KCNMB4 knockdown group (n=6 each) . The cardiac function was determined by echocardiography, left ventricular hemodynamics were measured invasively, renal sympathetic nerve activity (RSNA) was recorded at 6 weeks after coronary artery ligation or sham operation. The contents of norepinephrine (NE) and N-terminal pro-B-type natriuretic peptide (NT-proBNP) in plasma were determined by enzyme-linked immunosorbent assay. The protein and mRNA expression of KCNMB4 in PVN were measured by immunofluorescence staining, Western blot, and real-time PCR, mRNA expression of BKCa in PVN was detected by real-time PCR.@*Results@#Compared with the sham operation group, the cardiac function of the heart failure group was significantly reduced (P<0.05) , and the plasma NE and the serum NT-proBNP were significantly elevated (P<0.05) . The protein and mRNA expression of KCNMB4 in PVN were obviously down-regulated in CHF rats (P<0.05) . After perfusion of IBTX or KCNMB4 knockdown by microinjection of rAAV2-KCNMB4 shRNA virus,right ventricular weight/body weight and lung weight/body weight ratio as well as left ventricular end-diastolic diameter were increased and left ventricular ejection fraction was decreased (all P<0.05) , the sympathetic driving indexes was increased in sham rats, changes of these parameters further aggravated in CHF rats (P<0.05) . KCNMB4 knockdown further downregulated protein expression in PVN of CHF rats.@*Conclusion@#Downregulation and blunted function of BKCa in PVN may contribute to sympathoexcitation and deterioration of cardiac function in rats with chronic heart failure.
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CD36, the major scavenger receptor, is intimately involved in the uptake of oxLDL in macrophages. To further study the function of CD36 in macrophages, we constructed CD36 gene silence cell lines (J774A.1) by lentivirus-mediated RNA interference technique, and analyzed the effect of CD36 in caveolin-1 protein expression. At first, 5 shRNA fragments were designed and synthesized according to the coding sequence (CDS) region of CD36 gene. Next, the CD36-shRNA was inserted into lentiviral vector to yield pLKO.1-CD36-shRNA plasmid. After DNA sequencing, the pLKO.1-CD36-shRNA plasmid and psiCHECK-II-CD36 were co-transfected into the 293T cells to screen the efficient CD36-shRNA. The efficient CD36-shRNA plasmid and the helper plasmid were co-transfected into the 293T cells to package the lentivirus, and then infected the J774A.1 cells. After screening by puromycin, CD36 gene silence cell lines (J774A.1) was established. Western blotting and confocal fluorescence microscopy results showed that the CD36 silencing efficiency in the gene silence cell line was 90%. Accompanied by a decrease in CD36 protein on cell surface, oxLDL binding to CD36 was significantly inhibited, indicating that the CD36 gene silence cell line is successfully established. Finally, the oxLDL stimulation and inhibitor experiments results showed that the CD36 knockdown significantly suppresses the phosphorylation of JNK and ERK, thereby inhibiting the oxLDL-induced caveolin-1 protein expression, demonstrating that CD36 modulates the caveolin-1 protein expression through the JNK/ERK-mediated signaling transduction.
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OBJECTIVETo screen the allergens in patients with allergic rhinitis (AR) in Yibin, Zhaotong and some areas of the Changjiang River, and to study the distribution characteristics of inhaled allergens. METHODSThe clinical data of 661 patients with allergic rhinitis who diagnosed by skin prick test from March 2013 to June 2015 in the First people's Hospital of Yibin City were studied.RESULTSIn 661 patients, the top 5 allergens and their positive rates of skin prick test were Dermatophagoides farinae (76.5%), house dust mites (72.01%), Roach (35.85%), cat hair (11.35%) and dog hair (8.62%) respectively. There were no statistical difference of the positive rates of allergens among different seasons (χ2=4.568, 0.206). With the increasing age of the patients, the positive rate of allergens was decreased. CONCLUSION The dominated allergens in patients with AR are dust mites, house dust mites and cockroaches in Yibin, Zhaotong and and some areas of the Changjiang River. The prevention and control of AR in children should be toke more attention.
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OBJECTIVE@#To investigate mRNA expression of GABAa receptor(GABAaR) subunits in the rat cochlear spiral ganglion neurons (SGN) and explore the effect of sodium salicylate (SS) on the expression of GABAaR subunits.@*METHOD@#The realtime fluorescent quantitative PCR (FQ-PCR) was used to detect mRNA expression of twelve GABAaR subunits in the newborn rat SGN and then investigate mRNA expression of GABAaR subunits after treatment with 5 mmol/L SS for 15 min, 30 min, 1 h, 3 h and 6 h in the primary culture SGN.@*RESULT@#(1) GABAaR subunits of α1-6, β1-3, and γ1-3 were detected in the SGN, and the expression of GABAaR subunits was lower than those in the cerebral cortex. In the subunit α family of GABAaR, the expression rank was α2>α3/α5>α4>a1>α6, and the expression of α3 and α5 had no difference (P>0. 05). In the subunit β family, the expression rank was β3>β2>β1. In the subunit γ family, the expression rank was γ1>γ2>γ3. (2) The expression of all subunits of GABAa receptor was obviously fluctuated excepting subunit α5 after treatment with SS. At 15 min post-SS, α1, α2 , β1 and γ1-3 were upregulated, and α3 was downregulated; At 30 min post-SS, α3, β1 and β3 were upregulated, and γ1 was downregulated; At 1 h post-SS, β2 was upregulated and γ3 was downregulated; At 3 h post-SS, β1 and β2 were upregulated, and α3 and γ2 were downregulated; At 6 h post-SS, αl, α3 ,β2, β3 and γ1 were upregulated, and α2, α4 and β1 were downregulated.@*CONCLUSION@#The mRNA of GABAaR was expressed in the rat SGN, and the expression of GABAaR subunits was lower in SGN than the cerebral cortex. SS could alter the GABAaR expression quantity in rat SGN; Most of the subunits expression were elevated obviously in the early post SS (15 min), followed by a slight fluctuation.
Subject(s)
Animals , Rats , Cells, Cultured , Cochlea , Cell Biology , In Situ Hybridization , Neurons , RNA, Messenger , Receptors, GABA-A , Metabolism , Sodium Salicylate , Pharmacology , Spiral GanglionABSTRACT
For dangerous limits of capacitor energy, according to national standards of medical device regulations' test requirement, we analysis it and list its test methods and requirement. According to capacitor energy's formation and characteristics, we put forward a simple method for its test and calculation.
Subject(s)
Electric Capacitance , Electrical Equipment and Supplies , Reference Standards , Equipment SafetyABSTRACT
AIM: Investigated the relationship between NF-κB activation and cell apoptosis in mouse macrophages treated with 7-ketochesterol (7-KC). METHODS: Cell apoptosis was detected by MTT assay, DNA fragmentation assay and flow cytometric analysis. NF-κB activation was detected by western blot and immunohistochemistry. Inhibitory assay was used to show the effect of the activation of NF-κB on the apoptosis induced by 7-KC. RESULTS: 7-KC inhibited macrophages proliferation, and then induced apoptosis, which is associated with NF-κB activation. Moreover, cell apoptosis with NF-κB activation was inhibited by pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-κB. CONCLUSION: 7-KC induced the activation of NF-κB and following cell apoptosis.