AIM To investigate the effects of the novel antihypertensive drug iptakalim hydrochloride on potassium currents in artery smooth muscle cells. METHODS The effects of iptakalim hydrochloride on potassium currents in smooth muscle cells derived from rat pulmonary arteries were observed by using patch clamp technique(whole cell recording) after application of the drug in the bath. RESULTS The potassium current-voltage curves ( I-U curves) of smooth muscle cells derived from normotensive rat pulmonary arteries were up-ward shifted by iptakalim hydrochloride(0.1,1,10 and 100 ?mol?L -1 ). Within 5 minutes after application of the drug, the current amplitude could increase to[(118.6?15.9)%, P

Objective: To investigate the effect of taurine on learning and memory impairment, cytokines secretion in rats intrahippocampally injected with ?-amyloid (A?) 1-40. Methods: SD rats were randomly divided into control group, A? injected group, taurine (0.3g/kg?d, 0.6g/kg?d) groups. The rats were fed with taurine for 7 days, and then subjected to bilateral intrahippocampus injection of A?1-40 or vehicle. Two weeks later, all rats performed Morris water maze test. The contents of IL-6, TNF-? were checked by way of radio-immunity assay for hippocampus samples. Results: Compared with A?model group, the escape latency and distance were significantly reduced in taurine (0.6g/kg?d) group; the ratio of swimming distance in the target quadrant to that in the whole pool of the probe trial; the content of cytokines of IL-6 and TNF-?in hippocampus were reduced significantly. Conclusion: Taurine can effectively attenuate the cognitive dysfunction caused by A?1-40 in rats. The reduced cytokines content in hippocampus might contribute to this effect.

AIM: To analyze the effect of pigment epithelium derived factor (PEDF) on nitrogen monoxide (NO) and expression of cysteine-containing, aspartate-specific proteases-3 (caspase-3) in retinal tissues of model rats with optic nerve crush injury.METHODS: A total of 60 SD rats were randomly divided into the blank control group, model group and PEDF group, with 20 rats in each group.Except the blank control group, the optic nerve crush injury rat models were established in the other groups, and left eyeballs were taken as samples.After successfully modeling, the model group were treated with intravitreal injection of 5μL of balanced salt solution while PEDF group were treated with intravitreal injection of 5μL of PEDF (0.2μg/μL).Two weeks later, the retinal tissues were collected, and changes of shape were observed under microscope after HE staining.The changes of NO level were measured by colorimetry assay, the expression of caspase-3 mRNA and caspase-3 protein was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blot.RESULTS: HE staining showed that retinal tissues of the blank control group arranged neatly and clearly.Retinal ganglion cells (RGCs) arranged in a monolayer, and cells were oval, uniform in size and distribution, the cell nuclei were clear, closely arranged, with clear boundaries.The retinal tissues of the model group were sparse in shape, RGCs showed vacuolar changes, the overall number of cells was reduced, and cell nuclei of residual RGCs showed pyknosis and uneven staining.RGCs in PEDF group were with slightly edema and arranged closely, and the degree of injury was significantly milder than that in the model group.Levels of Caspase-3 mRNA and protein and NO levels in the three groups showed the model group > PEDF group > blank control group (all P < 0.05).CONCLUSION: The application of PEDF can down regulate the expression of Caspase-3 and NO in rates with optic nerve injury and reduce RGCs injury.

OBJECTIVEHigh altitue pulmonary edema (HAPE) impacts seriously people's health at high altitude. Screening of susceptibility genes for HAPE will be used for the evaluation and protection of susceptible people.

METHODSWe performed a genome-wide association study (GWAS) using Affymetrix SNP array 6.0 in 23 HAPE patients and 17 healthy controls. GO and Pathway analysis softwares were used to analyze and draw gene network.

RESULTSThirty-nine SNPs were found to be significantly different between case and control groups (P < 10(-4)). GO and Pathway analysis of 27 genes around the 39 SNPs indicated that these genes mainly participate in the regulating of cell proliferation, regulation of nitrogen compound metabolic process and G-protein coupled receptor protein signaling pathway and so on.

CONCLUSIONIt suggests that these SNPs and genes found in this study may be associated with the susceptibility of HAPE.

Adult ; Altitude Sickness ; genetics ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Genetic Predisposition to Disease ; Genome-Wide Association Study ; Humans ; Hypertension, Pulmonary ; genetics ; Polymorphism, Single Nucleotide ; Young Adult

AIMTo establish the culturing of cortical, hippocampal and sympathetic ganglia neurons derived from neonatal rats and the whole-cell recording techniques of potassium and sodium currents in cultured neurons.

METHODSCortex, hippocampus and sympathetic ganglion were isolated from neonatal rats (1-3 days) and digested by trypsin (0.125%). The neurons were cultured in polylysine covered petri dish (O35 mm) with DEME medium(high glucose) for seven days for electrophysiological experiments.

RESULTSThe neurons cultured by this method were in good conditions and could be sealed easily in whole-cell recording. I(Na), I(A), and I(K) could be recorded respectively in cultured neurons by different recording paradigms in Ca2+ -free extracellular solution.

CONCLUSIONIt is a practicable and economical method for electrophysiological study of nervous system.

Animals ; Calcium Channels ; physiology ; Cells, Cultured ; Cerebral Cortex ; physiology ; Ganglia, Sympathetic ; cytology ; physiology ; Hippocampus ; physiology ; Membrane Potentials ; physiology ; Patch-Clamp Techniques ; Potassium Channels ; physiology ; Rats ; Rats, Wistar ; Sodium Channels ; physiology

OBJECTIVETo explore the possible pathophysiological process and mechanisms underlying the development and formation of high altitude pulmonary edema(HAPE) by observing the changes in contents of VEGF, TNF-alpha, IL-6 and NO in serum from the initiated and recovery of HAPE patients.

METHODSWe studied 10 HAPE patients in a Chinese population. The patients were divided into two groups including HAPE initiate group and the recovery group. Contents of VEGF, TNF-alpha, IL-6 and NO in serum of the two groups were determined to study the process of HAPE.

RESULTSVEGF levels in the HAPE initiate one and the recovery groups were (167.9 +/- 26.5 and 53.1 +/- 17.0 pg/ ml), respectively. There was a significant decrease of VEGF content in recovery one compared to the HAPE group. The same results for TNF-alpha were gained. The levels of TNF-alpha in recovery group was much lower than that in the HAPE initiate one. They were (29.2 +/- 6.8) pg/ml and (86.2 +/- 24.1) pg/ml, respectively. The contents of IL-6 in HAPE initiate group and the recovery group were (32.3 +/- 16.5) pg/ml and (12. 5 +/- 8.0) pg/ml, respectively. But no significance existed. The level of NO in HAPE initiate group was (33.8 +/- 3.3) micromol/L, and it remarkably increased to (74.1 +/- 6.2) micromol/L in recovery one.

CONCLUSIONVEGF, TNF-alpha, IL-6 and NO participated in the different aspects of the pathophysiological process and might have influence on HAPE.

Adult ; Altitude ; Altitude Sickness ; complications ; physiopathology ; Humans ; Interleukin-6 ; blood ; Male ; Nitric Oxide ; blood ; Pulmonary Edema ; blood ; etiology ; physiopathology ; Tumor Necrosis Factor-alpha ; blood ; Vascular Endothelial Growth Factor A ; blood

OBJECTIVEHigh altitude pulmonary edema (HAPE), a life-threatening disease, has no biological markers used for the routine prevention, diagnosis and treatment. The aim of this study was to identify serum proteins differentially expressed in patients with HAPE for discovering essential biomarkers.

METHODSA complete serum proteomic analysis was performed on 10 HAPE patients and on 10 high altitude and 11 sea level healthy people as control using two-dimensional gel electrophoresis, followed by matrix-assisted laser desorption/ionization mass spectrometry and peptide mass fingerprinting. Finally, two most significantly changed proteins were validated by enzyme-linked immunosorbent assay (ELISA).

RESULTSEight protein spots stained with differential intensity, respresenting 5 distinct proteins were identified in patients compared with healthy controls through analysis of these composite gels. Among them, four proteins, namely alpha 1-antitrypsin(alpha1-AT), Haptoglobin(Hp), apolipoprotein A-1 (apoA-1) and Complement C3 increased remarkably, while one protein, apolipoprotein A-IV (apoA-IV) decreased significantly. The variation of alpha1-AT and Haptoglobin, as detected by ELISA, was consistent with the results from proteomic analysis.

CONCLUSIONSIt is well known that Hp, alpha1-AT and complement C3 are associated with inflammation and apoA-1 and apoA-IV play important roles in lipid absorption, transport and metabolism. Therefore, the significant expression changes of Hp, alpha1-AT and complement C3 and apoA-1 and apoA-IV between HAPE patients and their corresponding healthy controls highlight the role of inflammatory response system and lipid metabolism system in the pathophysiology of HAPE.

Altitude ; Biomarkers ; blood ; Blood Proteins ; metabolism ; Case-Control Studies ; Electrophoresis, Gel, Two-Dimensional ; Enzyme-Linked Immunosorbent Assay ; Humans ; Peptide Mapping ; Proteome ; Pulmonary Edema ; blood ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

AIMTo investigate the possible involvement of glutamate(Glu) in the paraventricular nucleus (PVN) in the central regulation of baroreflex.

METHODSThe baroreflex was induced by intravenous injection of phenylephrine in conscious rats, and the extracellular concentration of Glu in the PVN region was measured by microdialysis and high performance liquid chromatography (HPLC) techniques. To determine whether the observed Glu release was involved in the baroreflex, NMDA and non-NMDA receptor antagonists, MK-801 and CNQX, were perfused in the PVN region during baroreflex.

RESULTSDuring baroreflex, the Glu concentration in the PVN region immediately increased to 384.82% +/- 91.77% of basal level (P < 0.01). (2) During baroreflex, direct perfusion of MK-801 and CNQX in the PVN were attenuated the increase of blood pressure and enhanced the decrease of HR (P < 0.01),resulting a significant increase in baroreflex sensitivity (P < 0.01).

CONCLUSIONGlutamate in PVN is involved in central regulation of baroreflex, which may inhibit baroreflex via ionothopic glutamate receptors.

6-Cyano-7-nitroquinoxaline-2,3-dione ; pharmacology ; Animals ; Baroreflex ; drug effects ; physiology ; Dizocilpine Maleate ; pharmacology ; Excitatory Amino Acid Antagonists ; pharmacology ; Male ; Paraventricular Hypothalamic Nucleus ; physiology ; Rats ; Rats, Wistar

Finding the novel drug from the effective components of traditional Chinese herbal medicine is a hotspot of the modern pharmacological research.Hyperoside (HYP) belongs to flavonoid glycosides,and it has various properties,such as anti-inflammation,anti-spasm,anti-diuretic,antitussive,lowering blood pressure,and lowering cholesterol effects as well as protective effects for the cardiac and cerebral blood vessels.The purpose of this study was to investigate the effects of HYP on inflammatory and apoptotic responses in vascular endothelial cells stimulated by lipopolysaccharide (LPS) and further to identify the possible mechanisms underlying these effects.In our study,human umbilical vein endothelial cells (HUVECs) were stimulated with 1 μg/mL LPS in the presence or absence of HYP (10,20 and 50 μrnol/L).Our results indicated that HYP alone exerted no cytotoxicity on HUVECs,while it had an up-regulatory effect on the viability of HUVECs induced by LPS in a dose-dependent manner;increased mRNA expression of IL-1β,IL-6,TNFα and iNOS induced by LPS was attenuated after treatment with HYP both in a dose-and time-dependent manner;LPS-induced HUVECs apoptosis and cleaved-caspase 8,9,3 were all significantly reduced by HYP.Furthermore,the possible pathway involved in apoptosis and inflammation by HYP was detected,and the results showed that when treated with HYP,LPS-induced mitochondrial membrane instability was significantly inhibited through up-regulation of Bcl-2 and down-regulation of Bax.Furthermore,the expression of TLR4 and the phosphorylation of Iκ Bα and p65 in LPS-treated cells were blocked by HYP.Our results suggested that HYP treatment prevented HUVECs from LPS-induced inflammation and apoptosis responses,which might be mediated by inhibiting TLR4/NFκB pathway.

OBJECTIVETo understand the genetic structures and variations of the superintegron (SI) in Vibrio cholerae isolated in the seventh cholera pandemic.

METHODSPolymerase chain reaction scanning and fragment sequencing were used. Sixty toxigenic V. cholerae O1 El Tor strains isolated between 1961 and 2008 were analyzed.

RESULTSSome variations were found, including insertions, replacements, and deletions. Most of the deletions were probably the result of recombination between V. cholerae repeat sequences. The majority of the variations clustered together. The SIs of the strains isolated in the 1960s and 1970s showed more diversity, whereas SI cassette variations in strains isolated in the 1990s and after were lower, with ∼24 kb signature sequence deletion. This indicates the predominant SI in the host during the epidemic in the 1990s and after. The insertion cassettes suggested the mobilization from the SIs of other V. cholerae serogroups and Vibrio mimicus.

CONCLUSIONThe study revealed that structural variations of SIs were obvious in the strains isolated in epidemics in different decades, whereas the divergence was based on syntenic structure of SIs in these El Tor strains. Also, the continuing cassette flows in the SIs of the host strains during the seventh cholera pandemics were displayed.

Cholera ; epidemiology ; microbiology ; Chromosomes, Bacterial ; genetics ; ultrastructure ; Cluster Analysis ; DNA, Bacterial ; genetics ; Electrophoresis, Gel, Pulsed-Field ; Gene Deletion ; Gene Flow ; Genetic Variation ; Humans ; Integrons ; genetics ; Mutagenesis, Insertional ; Open Reading Frames ; genetics ; Polymerase Chain Reaction ; Tandem Repeat Sequences ; Vibrio cholerae O1 ; classification ; genetics ; isolation & purification ; ultrastructure