Objective To investigate the interference effects of orexin A on cell proliferation of the insulin?secreting beta?cell line(INS?1 cells) through the orexin receptor 1(OX1R)and the AKT/PKB signaling pathway. Methods INS?1 cells were exposed to different concentrations of orexin A in vitro,and treated with OX1R antagonist(SB334867),PI3K antagonist(wortmannin),or AKT antagonist(PF?04691502). The INS?1 cell proliferation and apoptosis,insulin secretion,OX1R protein activity and AKT phosphorylation level were determined. Results Orexin A(10-10 to 10-6 mol/L)stimulated the proliferation and activation of INS?1 cells,prevented apoptpsis,and increased insulin secretion. Additionally,AKT phosphorylation was stimulated by orexin A(10-10 to 10-6 mol/L). The OX1R antagonist SB334867(10-6 mol/L),the PI3K antagonist wortmannin (10-8 mol/L)and the AKT antagonist PF?04691502(10-6 mol/L)weakened the effects of orexin A. Conclusion Orexin A activated the AKT sig?naling pathway through the mediation of orexin A?OX1R,and promoted cell proliferation in INS?1 cells.

AIMTo investigate the changes of large-conductance calcium-activated potassium channels (BKCa, MaxiK) during aging and relations between the changes and blood pressure.

METHODSMale spontaneously hypertensive rats (SHR) aged 9, 15, 21, 27, 33 weeks (the number of each weeks SHR was 4) were selected as hypertension group rats, corresponding gender, weeks and number Wistar-Kyoto rats (WKY) as control group rats. Blood pressure of abdominalis aorta of each weeks SHR and WKY were measured by BL-420F experimental system of biological function. The arteria mesenteric minor (AMM) were isolated in blunt dissection method. The vascular smooth muscle cells (VSMCs) of AMM were isolated with prolease. The potassium current, the current after BKCa were blockaded by Tetraethylammonium (TEA) and the capacitance of membrane (Cm) of VSMCs of AMM were recorded with using whole cell patch clamp, and calculated the BKCa current and the BKCa current density. Probe the correlation of the changes of BKCa current density with MABP during aging.

RESULTSThe potassium current density and BKCa current density of VSMCs of AMM of SHR were decreasing during aging, however, the changes of WKY had no statistically significance (P > 0.05). The BKCa current density was extremely correlative with MABP in SH R (the values of r were -0.7174), in WKY, the BKCa current density was correlative with MAB P r = -0.4832.

CONCLUSIONBKCa current and current density attenuate with aging, the level of blood pressure is response of the attenuated degree. The BKCa current density is extremely correlative with the blood pressure.

Aging ; physiology ; Animals ; Blood Pressure ; physiology ; Cell Membrane ; physiology ; Hypertension ; metabolism ; physiopathology ; Large-Conductance Calcium-Activated Potassium Channel alpha Subunits ; Large-Conductance Calcium-Activated Potassium Channels ; metabolism ; physiology ; Male ; Membrane Potentials ; physiology ; Mesenteric Arteries ; cytology ; Muscle, Smooth, Vascular ; cytology ; metabolism ; physiology ; Patch-Clamp Techniques ; Potassium Channels ; metabolism ; physiology ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY

OBJECTIVE: To investigate characteristics of molecular etiology of children with profound sensorineural hearing loss in Hubei province, and to provide reference for deafness treatment and genetic counseling. METHOD: Three hundred and six children with profound sensorineural hearing loss in Hubei province were enrolled, their genomic DNA were extracted from peripheral blood and a deafness gene test chip was used to screen nine hot spot mutation in the GJB2, GJB3, SLC26A4, and mitochondria 12SrRNA gene. All patients with SLC26A4 gene mutation were given temporal bone CT scan. RESULT: One hundred and thirty-two (43.14%) out of 306 children were found carrying at least one pathogenic gene mutation. The mutation rates of GJB2, SLC26A4 and mitochondria DNA 12SrRNA gene were 29.41% (90/306), 13.72% (42/306) and 0.65% (2/306), respectively. None out of 306 children was detected GJB3 gene mutation. Thirty-six patients carrying SLC26A4 gene mutation were detected enlarged vestibular aqueduct by CT scan. CONCLUSION Mutations of GJB2 and SLC26A4 gene are two major pathogenic gene for genetic hearing loss in children. 235delC mutation is the main mutation type, followed by IVS7-2A> G mutation type. The screening of SLC26A4 gene common mutations contribute to the diagnosis of enlarged vestibular aqueduct syndrome.
Adolescent ; Child ; Child, Preschool ; China ; Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; Deafness ; genetics ; Female ; Genetic Testing ; Humans ; Infant ; Male ; Membrane Transport Proteins ; genetics ; Mutation ; Oligonucleotide Array Sequence Analysis ; Sulfate Transporters

Objective To evaluate calcium deficiency and demand in pre-puberty twins and to analyze the heritability of serum bone alkaline phosphatase(BALP).Methods A total of 73 pairs of twins aged 9-16 years were examined by BALP test.Microsatellite polymorphism Was used To diagnose the zygosity of twins,while both intraclass correlation coefficient method and Christian formula were perfortled to investigate heritability of serum BALP.Results The results of zygosity diagnosis displayed that 34 pairs of twins were monozygotic(MZ)twins and 39 pairs were dizygotic(DZ)twins.97.9％of the subjects appeared unusual in the activity of BALP,with activity of BALP>250 U/L in 43.1％of subjects and 200-250 U/L in 54.8％of the subjects.The intake of calcium Was unsatisfied in 48.4％of the boys and 39.0％in girls.Less than 10.0％ of the Sllbiects were satisfied with the intake of calcium in each age group while over 45.0％of the snbjects in 10-13 age group were deficient in calcium.Differences between the means of BALP in different sex groups(t=1.633,P=0.105)and different age groups(F=0.323,P=0.924)were not statistically significant.Heritability analysis displayed that intra-class variation.Inter-class variation.Intra-class correlation coefficient were 191.54,1462.22,0.77 in MZ twins,and those were 491.03,1475.57,0.50 in DZ twins respectively with the heritability of BALP activity as 0.54.Conclusion Calcium deficiency Is commonplace in pre-puberty twins.Our data showed that the BALP activity was influenced both by genetic(54％)and environmental(46％)factors.

Objective To investigate influencing factors of the first cases of varicella outbreaks in primary school in Pudong New Area of Shanghai and to provide scientific evidence for effective prevention and control measures against it. Methods The first 80 cases of varicella outbreaks in primary school in Pudong New Area were included for the case-control study.Immunization history, outing activities and information of out-of-school training were gathered.Univariate and multivariate conditional logistic regression analysis were adopted for data analysis. Results Multivariate analysis revealed that varicella vaccination period(OR=0.08, 95%CI:0.01-0.44)was the protective factor for the first case in a varicella outbreak, while weekly frequency of visits to large commercial/supermarkets (OR=4.79, 95%CI:1.86-12.34)and weekly attendance at out-of-school training(OR=1.84, 95%CI:1.01-3.35)were risk factors of the disease. Conclusion It is important to promote the current varicella vaccine immunization strategy, to strengthen the disinfection and health education of large commercial/supermarkets and to strengthen infectious disease management, disease prevention guidance and supervision in out-of-school training institutions.

OBJECTIVETo investigate whether adiponectin plays a role in the protection of myocardium in the rat myocardial ischemia preconditioning (IPC) model.

METHODInfarct size was measured by Masson's Trichrome staining, the expression of protein and mRNA of adiponectin at 0, 6, 12 and 24 h after IPC was examined by immunohistochemistry and quantitative real time RT-PCR, plasma levels of adiponectin at above mentioned four time points after IPC were detected by ELISA in IPC and MI rats.

RESULTInfarct size was smaller in IPC than in MI rats (20% ± 2% vs. 31% ± 3%, P < 0.05). The expression of adiponectin mRNA at 6 h and 12 h after IPC was 2.2 and 2.1 times higher than in Sham rats at respective time points (P < 0.05). Immunohistochemistry staining evidenced increased adiponectin expression in the ischemic area and weak expression of adiponectin in non-ischemic area (P < 0.05). Compared to the sham group, the plasma level of adiponectin increased significantly at 0, 6 and 12 h after IPC (0 h: 7.40 ± 0.47 vs. 10.90 ± 1.74; 6 h: 8.18 ± 1.41 vs. 10.98 ± 1.74; 12 h: 6.97 ± 1.02 vs. 9.31 ± 0.96, P < 0.05).

CONCLUSIONIPC reduced infarction size, upregulated the myocardial expression of adiponectin at mRNA and protein levels, and increased plasma adiponectin concentration, suggesting that the adiponectin may play a critical role in the protective effect of IPC.

Adiponectin ; metabolism ; Animals ; Ischemic Preconditioning, Myocardial ; Male ; Myocardial Infarction ; metabolism ; prevention & control ; Myocardial Ischemia ; metabolism ; prevention & control ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley

Adult ; Culture Media ; classification ; Female ; Humans ; Male ; Mycoplasma Infections ; microbiology ; Mycoplasma hominis ; growth & development ; Ureaplasma Infections ; microbiology ; Ureaplasma urealyticum ; growth & development ; Urethra ; microbiology ; Vaginal Smears

OBJECTIVEThe present study was to explore signaling mechanisms underlying nicotine-induced inhibition of large-conductance calcium-activated potassium channels (BK(Ca)).

METHODS8 week male Wistar rats were divided randomly into saline group and nicotine group and received respectively injection with saline or nicotine (Sigma, Shanghai, China) at 2 mg/(kg x d) for 21 days. Coronary vascular smooth muscle cells were dissociated enzymatically. Dissociated smooth muscle cells were interfered with CPT-cAMP (100 micromol/L) or forskolin (10 micromol/L). The signal channel open dwell-time (To), close dwell-time (Tc) and open probability (Po) were recorded.

RESULTSCPT-cAMP or forskolin significantly prolonged To, shorten Tc and increased Po in saline group (P < 0.01). But in nicotine group To, Tc and Po did not been changed.

CONCLUSIONThis phenomenon may serve as a physiological mechanism that nicotine inhibits BK(Ca) channel activity to increase via cAMP/PKA-dependent pathway.

Animals ; Arteries ; cytology ; drug effects ; metabolism ; Coronary Vessels ; cytology ; drug effects ; metabolism ; Large-Conductance Calcium-Activated Potassium Channels ; metabolism ; Male ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Nicotine ; pharmacology ; Patch-Clamp Techniques ; Rats ; Rats, Wistar ; Signal Transduction

OBJECTIVETo develop a fast, high-throughput screening method with suspension array technique for simultaneous detection of biothreat bacteria.

METHODS16 S rDNA universal primers for Bacillus anthracis, Francisella tularensis, Yersinia pestis, Brucella spp.and Burkholderia pseudomallei were selected to amplify corresponding regions and the genus-specific or species-specific probes were designed. After amplification of chromosomal DNA by 16 S rDNA primers 341A and 519B, the PCR products were detected by suspension array technique. The sensitivity, specificity, reproducibility and detection power were also analyzed.

RESULTSAfter PCR amplification by 16 S rDNA primers and specific probe hybridization, the target microorganisms could be identified at genus level, cross reaction was recognized in the same genus. The detection sensitivity of the assay was 1.5 pg/microl (Burkholderia pseudomallei), 20 pg/microl (Brucella spp.), 7 pg/microl (Bacillus anthracis), 0.1 pg/microl (Francisella tularensis), and 1.1 pg/microl (Yersinia pestis), respectively. The coefficient of variation for 15 test of different probes was ranged from 5.18% to 17.88%, it showed good reproducibility. The assay could correctly identify Bacillus anthracis and Yersinia pestis strains in simulated white powder samples.

CONCLUSIONThe suspension array technique could be served as an opening screening method for biothreat bacteria rapid detection.

Bacillus anthracis ; isolation & purification ; Bioterrorism ; prevention & control ; DNA Primers ; DNA, Bacterial ; analysis ; Francisella tularensis ; isolation & purification ; Oligonucleotide Array Sequence Analysis ; methods ; Polymerase Chain Reaction ; methods ; RNA, Ribosomal, 16S ; genetics ; Yersinia pestis ; isolation & purification

OBJECTIVEThe purpose of this study was to assess weather the immortalized mouse brain endothelial cell line Bend.3 displays the comparative barrier characteristics as the primary brain microvascular endothelial cells (BEMC).

METHODSImmortalized mouse brain endothelial cell line, Bend.3 cells were cultured in transwell inserts and their restrictive characteristics were assessed by transendothelial electrical resistance (TEER) and horseradish peroxidase (HRP) permeability assays. Western blot and direct fluorescent staining methods were used to detect the tight junction protein expression and F-actin distribution.

RESULTSThe TEER in Bend.3 cells increased with the prolonged culture time and increased to 82.3+/-6.0 Omega cm2 10 days after culture, which was significantly higher than that 3 days after culture (37.3+/-3.1 Omega cm2; P<0.05). There were significant differences in the permeability rates for HRP 3 and 10 days after culture (4.3+/-0.20)% vs (2.2+/-0.05)% (P<0.05). Western blot indicated high level expression of tight junction proteins occludin and ZO-1 in Bend.3 cells 10 days after culture. F-actin was visualized around the cell membrane and presented scrobiculate linear fluorescence 10 days after culture.

CONCLUSIONSBend.3 cells have similar barrier characteristics to BEMC, and their barrier function may reach to the best effect 10 days after culture.

Actins ; analysis ; Animals ; Blood-Brain Barrier ; Cell Line ; Electric Impedance ; Endothelial Cells ; metabolism ; Horseradish Peroxidase ; metabolism ; Membrane Proteins ; analysis ; Mice ; Phosphoproteins ; analysis ; Zonula Occludens-1 Protein