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Objective: Many studies have shown that respiratory syncytial virus persistent infection may be the main cause of chronic respiratory pathology. However, the mechanism is unclear. Cystic fibrosis transmembrane conduction regulator (CFTR) is an apical membrane chloride channel, which is very important for the regulation of epithelial fluid, chloride ion, and bicarbonate transport. CFTR dysfunction will lead to changes in bronchial secretions and impair mucus clearance, which is related to airway inflammation. In our previous study, we observed the down-regulation of CFTR in airway epithelial cells in respiratory syncytial virus (RSV) infected mouse model. In this study, we further investigated the expression and function of CFTR by constructing an airway epithelial cell model of RSV persistent infection. Methods: 16HBE14o- cells were infected with RSV at 0.01 multiplicity of infection (MOI). The expression of CFTR was detected by real-time RT-PCR, immunofluorescence, and Western blotting. The intracellular chloride concentration was measured by N-(ethoxycarbonylmethyl)-6-methoxyquinolium bromide (MQAE) and the chloride current was measured by whole-cell patch clamp recording. Results:16HBE14o-cells infected with RSV were survived to successive passages of the third generation (G3), while the expression and function of CFTR was progressively decreased upon RSV infection from the first generation (G1) to G3. Exposure of 16HBE14o-cells to RSV led to the gradual increase of TGF-β1 as well as phosphorylation of Smad2 following progressive RSV infection. Disruption of TGF-β1 signaling by SB431542 prevented Smad2 phosphorylation and rescued the expression of CFTR. Conclusion:RSV infection can lead to defective CFTR function in airway epithelial cells, which may be mediated via activation of TGF-β1 signaling pathway.
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Objective To explore the effect of expression of protein kinase C receptor 1(RACK1) induced by lipopolysaccharide (LPS) on Sonic hedgehog(SHH) signaling pathway in rat puhnonary microvascular endothelial cells (RPMVEC).Methods The healthy male SPF grade SD rat with 100-120 g body weight were gotten from the laboratory animal center of Anhui province.Using immunocytochemistry method,the expression of RACK1 protein in RPMVECs was detected,cultured RPMVECs were randomly divided into different groups as LPS dose-dependent group,SAG(smoothened Agonist,a SHH signaling pathway specific agonist) dose-dependent group,LPS time-dependent group,SAG time-dependent group and LPS+SAG group.In LPS dose-dependent groups,RPMVECs were cultured with 0.1,1,10 mg/L LPS for 8 h.In LPS time-dependent groups,RPMVECs were cultured with 10 mg/L LPS for 0,2,4,8,12,24 h.In SAG dose-dependent groups,RPMVECs were cultured with 0.1,1,10 μ mol/L for 8 h.In SAG time-dependent groups,RPMVECs were cultured with 1 μ mol/L SAG for 0,2,4,8,12,24 h.In LPS+SAG group,RPMVECs were cultured with 1 μ mol/L SAG 8 h after 10 mg/L LPS treatment for 1 h.In addition,blank group,LPS group and SAG group were set for control.Western blot were used to detect the level of RACK1 and RT-PCR were used to detect the expression of GLI-1 mRNA after intervention.Results Immunocytochemistry revealed that RACK1 were present in RPMVEC.1.In LPS dose-dependent groups (0,0.1,1,10 mg/L),the level of RACK1 elevated as LPS dose increased correspondingly with inter-group difference (P＜0.05);the relative expression levels of GLI-1 mRNA were (1.109 + 0.063),(1.039 + 0.135),(0.813 ± 0.066),(0.770 + 0.105),(1 mg/L vs.10 mg/L,P＞0.05;the rest P＜0.05).In LPS time-dependent groups,the relative expression level of RACK1 at 2 h (0.370 + 0.010) was higher than that at 0 h (0.329 ± 0.008),peaked at 12 h (1.296 ± 0.048),and compared with 0 h,there was significant differences (F=1 272.204,P＜0.05).The relative expression level of GLI-1 mRNA was decreased at 2 h (0.929 ±-0.007),and compared with 0 h(1.089 ± 0.042),there was significant differences (F=306.609,P＜0.05).2.In SAG dose-dependent groups,there was no significant difference in level of RACK1 between groups(all P＞0.05).The relative expression levels ofGLI-1 mRNA were (1.109 ± 0.063),(1.169 ± 0.052),(3.468 ± 0.128),(3.434 ± 0.054),(0 μ mol/L vs.0.1 μ mol/L and 1 μ mol/L vs.10 μ mol/L,P＞0.05,the rest P＜0.05).Among SAG time-dependent groups,there was no significant difference in levels of RACK1 protein(P＞0.05).The relative expression level of GLI-1 mRNA increased at 2 h (3.027 ± 0.065),and compared with 0 h (2.651 + 0.123),there was significant differences (F=132.841,P＜0.05).3.In LPS+SAG intervention groups,the expression of RACKI was lower than that in LPS group (0.831 ±0.040 vs.1.189 ± 0.149,P＜0.05),and the expression of GLI-1 mRNA was higher than that in LPS group (2.720 + 0.130 vs.0.796-4-0.082,P＜0.05).Conclusions The LPS up-regulates the expression of RACK1 in RPMVECs,and the activated SHH signaling pathway can down-regulate the expression of RACK1 induced by LPS in RPMVECs.
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Objective To observe the clinical efficacy of puncturing Tianshu (ST 25) alone in treating intractable constipation in the elderly.Method Totally 193 senile subjects with intractable constipation were randomized into group A of 57 cases, group B of 73 cases, and group C of 63 cases. Group A was intervened by acupuncture at Tianshu (ST 25), group B was by Lactulose oral solution, and group C was by retention enema. The clinical efficacies and effect-lasting time of the three groups were compared. Result The total effective rate was 89.3% in group A, versus 88.6% in group B and 92.1% in group C, and the between-group differences were statistically insignificant (P>0.05). Of the markedly effective and improved cases, the effect-lasting time in group A was significantly different from that in group B and C (P<0.01).Conclusion Acupuncture at Tianshu (ST 25) alone is an effective method in treating senile intractable constipation.
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Objective To establish an animal model of preeclampsia by injecting ultra-low-dose lipopolysaccharide (LPS) to rats in early pregnancy,and to lay the foundation for further study on mechanisms of preeclampsia.Methods Twenty-four pregnant rats were divided into six groups according to the random number table and were injected with LPS 0.3,0.5,0.7,1.0,2.0 μg/kg or saline 2 ml respectively through tail veins on day 5 of pregnancy.The differences in blood pressure,urinary protein and pathological changes in placenta among groups were compared to confirm the suitable dose of LPS for establishing preeclamptic model.Then another 19 pregnant rats were injected with the chosen dose of LPS slowly through tail veins on day 5 of pregnancy; 15 of which were chosen as model group; the other four were chosen as postpartum group.Three non-pregnant rats were as non-pregnant group.Besides,another 15 pregnant rats were injected with saline as pregnant control group.Systolic blood pressure,urinary protein excretion,placental weight,fetal weight,serum white blood cell counts,blood platelet counts,plasma anti-thrombin-Ⅲ content,D-dimer content were examined and compared among groups with one way analysis of variance; histopathologic studies were also done on the placentas,kidneys and aortas of the rats.Results (1) Placental weight of LPS 0.3 μg/kg group increased compared with control group.One pregnant rats(1/4) in LPS 1.0 μg/kg group and LPS 2.0 μg/kg group died on day 16 of pregnancy as a result of vaginal bleeding.Systolic blood pressure of LPS 0.5 μg/kg group rose steadily,while no significant changes were found in other groups.Urinary protein increased in all LPS groups,while urinary protein of LPS 0.7 μg/kg group and LPS 1.0 μg/kg group peaked on day 12 of pregnancy and then decreased; urinary protein of LPS 0.5 μg/kg group increased most significantly,and fetus in LPS 0.5,0.7 and 2.0 μg/kg groups had lighter body weight.So LPS 0.5 μg/kg was chosen as the suitable dose to establish preeclamptic model.(2)Compared with pregnant control group,model group had higher systolic blood pressure [(124.89±1.79) mm Hg vs (119.02±1.80) mm Hg,LSD test,P=0.03] from day 6 of pregnancy,more urinary protein [(2.02±0.29) mg vs (1.11±0.18) mg,LSD test,P=0.00] from day 9 of pregnancy,more absorbed embryos [3.6％ (7/194) vs 0.0％ (0/200),Fisher exact test,P=0.01] at day 20 of pregnancy,higher incidence of placenta bleeding [4.1％ (8/194) vs 0.0％ (0/200),Fisher exact test,P=0.00] and fetal growth restriction [13.9％ (27/194) vs 6.0％ (12/200),X2=6.92,Fisher exacttest,P=0.01].Model group showed more inflammatory cells infiltration in the placenta,more glomerular mesangial cells,swelling and desquamated of renal tubular epithelial cells compared to control group.Blood pressure and urinary protein of the model group recovered to the baseline at the sixth day of postpartum,and no changes in blood pressure and urinary protein were found in non-pregnant rats.Conclusions Injection of LPS 0.5 μg/kg on day 5 of pregnancy through tail veins could induce the clinical symptoms of preeclampsia in rats,which might be an ideal model for further preeclampsia research.
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<p><b>OBJECTIVE</b>Antibacteria activity of compounds from Puraboeo ruescens and Lysionotus pauciflorus was assayed.</p><p><b>METHOD</b>Disc diffusion was used to isolate compounds in vitro and berberine was positive control. The value of IC50 was assayed by the method of liquid culture. All kinds of chromatography were used to isolate the chemical constituent and structure was identified by MS and NMR spectroscopy.</p><p><b>RESULT</b>Eight compounds were isolated and identified as beta-sitosterol (1), E-3,4-dihydroxy cinnamic acid (2), barbinervic acid (3), 3beta,19alpha-dihydroxy12-en-28-ursolic acid (4), 28-O-beta-D-glucopyranosyl pomolic acid (5), 5,7-dihydroxy-6,8,4'-trimethoxy flavone (6), 5, 6, 4'-trihydroxy-7,8-dihydroxy flavone (7), 5-hydroxy-6,8,4'-trimethoxy flavone-7-O-beta-D-glucopyranosyl (8). Compound 3, 4 and 6 had activity against SA, MRSA and ESBLs respectively. Compound 3 showed (IC50 = 0.098 g x LU(-1), IC50 = 0.27 g x L(-1)) against SA and ESBLs-SA respectively. Compound 4 (IC50 = 0.130 g x L(-1)) was best to against MR SA.</p><p><b>CONCLUSION</b>Compound 1 - 5 were isolated from this plant for the first time. Compound 7 and 8 was isolated from Gesneriaceae for the first time.</p>
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Anti-Bacterial Agents , Pharmacology , Bacteria , Drugs, Chinese Herbal , Pharmacology , Magnoliopsida , ChemistryABSTRACT
<p><b>OBJECTIVE</b>The alpha-glucosidase inhibitory compounds were searched from Forsythia suspense.</p><p><b>METHOD</b>The active compounds were isolated by the method of bioassay-guided in vitro and column chromatographic techniques. The structures of compounds were identified by MS and NMR spectroscopy. The inhibitory kinetics of the isolated compounds were investigated.</p><p><b>RESULT</b>The ethyl acetate extract showed the strongest inhibitory activity, and five active compounds were isolated from this extract. The IC50 value of compound 1-5 were all lower than that of acarbose as positive control. Compound 1, the mixture of 1, 2 and 4 all showed noncompetitive type model on alpha-glucosidase.</p><p><b>CONCLUSION</b>Compound 1-3 as the inhibitors of alpha-glucosidase were reported for the first time. Compound 3 was isolated from the genus for the first time.</p>
Subject(s)
China , Enzyme Inhibitors , Chemistry , Forsythia , Chemistry , Glycoside Hydrolase Inhibitors , Kinetics , Plant Extracts , Chemistry , Plant Leaves , Chemistry , alpha-Glucosidases , ChemistryABSTRACT
OBJECTIVE: To analyze the volatile components in the Eriobotrya japonica by headspace solid-phase microextraction-gas chromatography-mass spectrometry(SPME-GC-MS) and to optimize the operating conditions for SPME.METHODS: DB-5 MS elastic silica capillary column(30.0 m?250 ?m?0.25 ?m) was used as the chromatographic column with helium of high purity(99.999%) as carrier gas at a flow rate of 1.0 mL?min-1.The injector temperature was 250 ℃;MS condition: ionization mode was EI with electron energy of 70 eV and the mass scan range of 30～440 amu.RESULTS: After headspace absorption for 30 min at 80 ℃ using 65 ?m polydimethylsiloxane-divinyl-benzene extraction head followed by desorption for 1 min at 250 ℃,the E.japonica sample was separated and identified by GC-MS analysis.A total of 34 peaks were obtained and 29 components were identified which accounted for 97.42% of the total peak area,topping the list were benzaldehyde and 4-methoxybenzaldehyde.CONCLUSION: HS-SPME-GC-MS can be used for the rapid analysis of the volatile components in E.japonica.
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OBJECTIVE:To analyze the components of the volatile oil in different parts of Lonicera japonica Thunb.from Henan province.METHODS:The volatile oil was extracted from the buds,leaves,stems of L.japonica Thunb.by solid-phase micro-extraction,and the chemical compositions were identified by GC-MS combined with Kvotas retention index.The relative percentage of each constituent was determined by GC area normalization method.RESULTS:Thirty-nine compounds were identified from different parts of L.japonica Thunb.from Henan province,and seven of which were mutual in the buds,leaves,stems of L.japonica Thunb..CONCLUSION:This study serves as a scientific basis for the further development and utilization of L.japonica Thunb..
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OBJECTIVE:To analyze the fatty acids in rinds or seeds of Pittosporum tobira.METHODS:The oil from rinds or seeds of Pittosporum tobira was extracted with organic solvent,and the fatty acids were methyl-esterified with KOH-NaOH solution and identified by GC-MS.RESULTS:13 fatty acids were identified from rinds and seeds of Pittosporum tobira,mostly the palmitic acid and oleic acid.CONCLUSION:Both rinds and seeds of Pittosporum tobira can be exploited as an edible oil for health care.