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Aim To investigate the impact and mechanism of Weichang'an Pill(WCA),its ethanol extract(EE),water extract(WE),and active ingredients on the contraction of isolated rat ileum smooth muscles induced by acetylcholine(ACh). Methods In vitro tissue bath experiment,WCA,EE,WE,or their active ingredients were added under the action of ACh,and then the contraction tension of isolated ileum smooth muscle from rats was recorded. The binding affinity ofthe active ingredients to the muscarinic acetylcholine M3 receptor was explored by molecular docking. Results WCA,EE,and WE were able to considerably inhibit the excitatory contraction of the ileal smooth muscles induced by ACh. Costunolide,dehydrocostus lactone,santalol,muscone,emodin,chrysophanol,physcion,crotonoside,magnolol,and honokiol were also significantly effective against ACh-induced ileal smooth muscle contraction. Conclusions WCA,EE,WE,and their active ingredients may help to promote intestinal smooth muscle relaxation by blocking the binding of the M3 receptor on the membrane of ileal smooth muscle with ACh.
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Aim To investigate the effect of XNST and its monomeric components on the barrier structure and tight junction protein expression of brain microvascular endothelial cells damaged by oxygen glucose deprivation/reoxygenation (OGD/R) and the possible mechanism. Methods The mouse brain microvascular endothelial cell line bEnd. 3 was inoculated in the upper layer of the Transwell chamber to establish an OGD/R damage model, and the effect of the drug on the integrity of the endothelial cell barrier was investigated by the transmembrane resistance value and fluorescein-so-dium transmittance. Claudin-5 immunofluorescence staining was used to observe the changes of tight junction structure between endothelial cells. RT-PCR and Western blot were employed to detect mRNA and protein expression levels of tightly linked proteins Claudin-5 , Occludin, ZO-1. Western blot was applied to detect the expression levels of MAPKs (JNK, p38, ERK) , I kappa B a, I kappa B kinase phosphorylated protein expression, and Western blot and immunofluorescence were utilized to detect NF-K.B/p65 nucleation expression. Results XNST and its three monomers could significantly increase endothelial cell resistance and de- crease fluorescein-sodium transmittance. Claudin-5 fluorescence staining showed that the tight junction between cells in the model group was significantly damaged , while XNST and its monomer components could significantly improve its tight structure. RT-PCR and Western blot results showed that it could significantly upregulate the expression of mRNA and protein of Claudin-5, Occludin and ZO-1, and further study on the mechanism showed that XNST and its monomer components could significantly inhibit the phosphoryla-tion of JNK, p38 and ERK, inhibit the phosphorylation of I kappa B a and I kappa B kinases, and significantly inhibit the nuclear translocation of NF-KB/p65. Conclusion Both XNST and its monomeric components can exert cerebroprotective effects by increasing the tight junction structure between cells to promote barrier integrity, and the mechanism may be related to inhibition of NF-kB and MAPKs signaling pathway activation.
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Objective:To investigate the effect of salvianolate lyophilized injection and Xueshuantong injection (lyophilized) on the permeability of blood-brain barrier (BBB) via inhibition of metallomatrix protease（MMPs） in cerebral ischemia/reperfusion injury rats. Method:The focal cerebral ischemia/reperfusion model in rats was built by middle cerebral artery occlusion/reperfusion (MCAO/R) technique. Male Wistar rats were randomly divided into sham operation group, ischemia/reperfusion (I/R) group, edaravone (Eda, 6 mg·kg-1) group, salvianolate lyophilized injection (SLI, 21 mg·kg-1) group, Xueshuantong (XST, 100 mg·kg-1) group and SLI combined with XST (SLI+XST, 21 mg·kg-1+100 mg·kg-1) group. Drugs were injected via tail vein for 2 d, while sham group and I/R group were injected with the same amount of normal saline. Neurological deficit score, hematoxylin-eosin (HE) staining and Nissl staining were assessed 2 d after MCAO/R. The permeability of BBB was observed by the leakage of IgG/CD31. The expressions of Claudin-5，Occludin，collagen-Ⅳ（Col- Ⅳ），Laminin，Fibronectin were observed by immunofluorescence staining，and MMP-2 and MMP-9 were observed by Western blot. Result:Compared with the I/R group, SLI group, XST group and SLI+XST group showed improvements in neurological deficit score, HE staining and Nissl staining. The leakage of IgG was alleviated; The positive expressions of Claudin-5，Occludin，Col-Ⅳ，Laminin，Fibronectin in ischemic penumbra were significantly up-regulated, while the expressions of MMP-2 and MMP-9 were down-regulated. The effect in improving SLI combined with XST was much better than a single factor. Conclusion:Salvianolate lyophilized injection and Xueshuantong injection (lyophilized) can alleviate cerebral ischemia/reperfusion injury and exert the synergistic effect when they are used in combination. The mechanisms might be associated with the improvement in the permeability of blood-brain barrier by inhibiting MMPs in cerebral ischemia/reperfusion injury rats.
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OBJECTIVE@#To investigate the ameliorate effect and underlying mechanism of Xueshuantong for Injection (Lyophilized, , XST) in streptozocin (STZ)-induced diabetic retinopathy (DR) rats.@*METHODS@#Diabetes mellitus (DM) model was induced by intraperitoneal (i.p.) injection of STZ (60 mg/kg) in Sprague-Dawley rats. Diabetic rats were randomized into 3 groups (n=10) according to a random number table, including DM, XST50 and XST100 groups. XST treatment groups were daily i.p. injected with 50 or 100 mg/kg XST for 60 days, respectively. The control and DM groups were given i.p. injection with saline. Blood glucose level and body weight were recorded every week. Histological changes in the retina tissues were observed with hematoxylin-eosin staining. Apoptosis and inflammation related factors, including cleaved caspase-3, glial fifibrillary acidic protein (GFAP), tumor necrosis factor-α (TNF-α) and intercellular cell adhesion molecule-1 (ICAM-1) were detected by Western blot or real-time polymerase chain reaction. Then, the levels of advanced glycation end product (AGE) and its receptor (RAGE) were investigated. Tight junctions proteins (Zonula occludens-1 (ZO-1), Occludin and Claudin-5) of blood-retinal barrier were detected by Western blot. The levels of retinal fifibrosis, transforming growth factor-β1 (TGF-β1)-Smad2/3 signaling pathway were evaluated at last.@*RESULTS@#There was no signifificant difference in the body weight and blood glucose level between XST and DM groups (P>0.05). Compared with the DM group, XST treatment signifificantly increased the retinal thickness of rats (P<0.05 or P<0.01), and suppressed cleaved caspase-3 expression (P<0.01). XST increased the protein expressions of ZO-1, Occludin and Claudin-5 and decreased the mRNA expressions of matrix metalloproteinase 2 (MMP-2) and MMP-9 (P<0.05 or P<0.01). Moreover, XST signifificantly reduced the productions of AGE and RAGE proteins in the retina of rats (P<0.05 or P<0.01), suppressed the over-expression of TNF-α, and decreased the elevated level of ICAM-1 in retina of rats (P<0.05 or P<0.01). XST signifificantly reduced the levels of α-smooth muscle actin (α-SMA), connective tissue growth factor (CTGF), TGF-β1 and phosphorylation of Smad2/3 protein in rats (P<0.05 or P<0.01).@*CONCLUSIONS@#XST had protective effects on DR with possible mechanisms of inhibiting the inflammation and apoptosis, up-regulating the expression of tight junction proteins, suppressing the productions of AGE and RAGE proteins, and blocking the TGF-β/Smad2/3 signaling pathway. XST treatment might play a role for the future therapeutic strategy against DR.
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OBJECTIVE@#To test the role of psoralidin in human liver cancer HepG2 cells in vitro.@*METHODS@#Cell viability was assessed by methylthiazolyldiphenyl-tetrazolum bromide assay and apoptotic cells were labeled by annexin V then sorted by flow cytometry. Protein expressions of caspase-3, caspase-8, caspase-9, Bax, Bid, Bcl-2, Bcl-xL and p53 were examined by western blot while activity of caspase-3, -8 and -9 were also determined.@*RESULTS@#Psoralidin reduces cell viability greatly in a time dependent manner (64%, 40%, 21%, 12% at 2, 6, 24 and 48 h treatment with 64 μmol/L psoralidin respectively) and up-regulates activities of caspase-3, -8 and -9 in a concentration dependent manner (between 4 to 64 μmol/L). Psoralidin also increases the expression of pro-apoptosis genes Bax, Bid and p53 while decreases the expression of pro-survival genes Bcl-2 and Bcl-xL, both in a concentration dependent manner between 4 and 64 μmol/L (P<0.05 at 16 and 64 μmol/L). Caspase-3 inhibitor (Ac-DEVD-CHO at concentrations between 10 to 20 μmol/L), p53 inhibitor (pifithrin-α at 5 μmol/L) and cyclosporin A can attenuate the apoptotic effect of psoralidin.@*CONCLUSION@#The cytotoxic role of psoralidin might work through both intrinsic and extrinsic apoptotic pathway.
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Objective to compare and contrast the diagnosis results on 2D and 3D Ultrasonogrpahy against MRI. Method A 2D Ultrasonography was applied during a conventional prenatal sonography checking with a 3D Sonography assessment subsequently conducted to follow up on 49 fetus suspected of brain malformation.Furthermore, a MRI scan was taken within 24 hours after the 3D Sonography checking in our hospital as a final test. Data collections from all three assessments were completed, and an analysis of the comparisons of these three methods were done. Results Among these 49 fetus with confirmed or suspected brain malformation, there were two cases of misdiagnoses of Dandy-Walker Malformation assessed by 2D sonography, with a misdiagnosis rate of 40％, (P ＜ 0.05) indicating a statistically significant result; misdiagnosis rate of Fetal Ventriculomegaly and Isolated Broadening Posterior Fossa were calculated as 26.7％ and 33.3％ respectively (P ＜0.05), a statistically significant result. Overall, there were two cases with Cerebellum Malformation, from in which one case was identified by MRI, and the other one was misdiagnosed, with a misdiagnosis rate of 50.0％.In total, there were 2 cases of Holoprosencephaly, in which one was identified by Prenatal MRI, and the other was misdiagnosed (P ＜ 0.05), a statistically significant result. Conclusions All three assessments of 2D ultrasonography, 3D ultrasonography and MRI have their own advantages and disadvantages. In short, 2D Sonography is suggested to be applied for screening out cases with brain malformation, together with 3D Sonography as a complementary assessment. MRI can also be an effective and significant complement for sonography in completing and readdressing the final ultrasonic results.
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Objective To explore the clinical application value of rapid detection method for respiratory tract influenza virus and mycoplasma pneumonia(MP).Methods 386 cases were selected from patients with respiratory infections in our hospital from Janu-ary 2011 to December 2015 in the study,nasopharyngeal secretions A,B influenza virus were detected by gold immunchromato-graphic assay,the Mycoplasma pneumoniae(MP)was detected by rapid culture,and pathogens-IgM were detected by immunofluo-rescence assay.In the end,the two rapid detection detection method were compared with immunofluorescence assay.Results 386 cases of respiratory tract infections patients were detected by immunofluorescence assay,54 cases were found influenza A virus in-fection,the positive rate was 13.59%,37 cases were found influenza B virus infection,the positive rate was 9.59%,61 cases were found MP infection,the positive rate was 15.80%.In detections of influenza A virus infection and influenza B virus infection,im-munchromatographic assay sensitivity were 20.37%,18.92% and specificity were 93.41%,92.77%.In detection of MP infection, rapid culture sensitivity was 37.70% and specificity was 89.85%.Conclusion Rapid detection of respiratory influenza virus,MP infection sensitivity is low,only as a supplementary means of indirect immunofluorescence assay clinically.
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Objective To explore the effects of angled abutments on the anterior maxilla implant restoration. Methods We analyzed the biomechanical properties of implants of different sizes (Φ3.5 mm,4.0 mm and 4.5 mm in diameter;L11.5 mm and L13 mm in length)after connecting different angled abutments (0°,10°,20°,and 30°) using finite element method.Results The stresses and strains of loading parts of restorations increased and their distribution became more concentrated as the angle of abutment increased.Cortical bone of Φ3 .5 implants with smaller angle (10°or less)andΦ4.0 implants with abutments had the risk of overpassing the bone elastic threshold when the angle approached 30°.However,the cortical bone elastic deformation was within a safe range at all angles inΦ4.5 group.Conclusion We should consider the diameter of the implant when selecting angled abutments.The angled abutments are not suitable for small diameter implants.The bite force should be under control when needed. The larger angled abutments can be applied in the standard and major diameter implants and it is necessary to avoid occlusal overloading.
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<p><b>OBJECTIVE</b>To construct the vectors of human glutathione S-transferase A1 (GSTA1), P1 (GSTP1), T1(GSTT1) genes and express in Escherichia coli (E. coli).</p><p><b>METHODS</b>Human GSTA1, GSTP1 and GSTT1 gene whole length cDNAs were amplified by RT-PCR and then subcloned into pET-28a(+) vectors. The proteins were expressed in E. coli BL21(DE3). After purified by Ni2+ affinity chromatography, the enzymatic activities of GSTs were measured with 1-chloro-2,4 -dinitrobenzene (CDNB) as substrate.</p><p><b>RESULTS</b>The correct GSTA1, GSTP1 and GSTT1 genes were cloned. And soluble GSTA1, GSTT1, GSTP1 proteins were expressed in E.coli. After purification, GSTA1, GSTT1 and GSTP1 showed good enzymatic activities, which were 17.55, 0.02, 18.75 μmol·min-1·mg-1, respectively.</p><p><b>CONCLUSION</b>The expression plasmids for GSTA1, GSTT1 and GSTP1 have been constructed and the recombinant proteins are expressed successfully.</p>
Subject(s)
Humans , DNA, Complementary , Genetics , Escherichia coli , Genetics , Genetic Vectors , Glutathione S-Transferase pi , Genetics , Glutathione Transferase , Genetics , Recombinant Proteins , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Sustained activation of β adrenergic receptor (βAR) leads to pathologic cardiac hypertrophy. However, the related mechanisms still remain unclear. In this study, we observe how N-acetylcysteine (NAC) affects the oxidative stress and calcium/calmodulin-dependent protein kinase II (CaMKII) expression in heart of isoproterenol (ISO)-stimulated rats, and investigate whether oxidative stress and CaMKII contribute to the development of sustained βAR-stimulated cardiac hypertrophy. Healthy male Wistar rats were randomly separated into 4 groups: control (CTRL), ISO-treated (ISO), control with NAC supplement (CTRL+NAC) and ISO-treated with NAC supplement (ISO+NAC) groups (6 rats in each group). Systolic blood pressure (SBP) was measured in awake rats with the tail-cuff method every week for two weeks. Heart weight/body weight ratio (HW/BW) and HE staining were used for the detection of myocardial hypertrophy. Myocardial mitochondrial reactive oxygen species (ROS) levels were measured by DCF fluorometry. The expressions of activated-CaMKII (p-CaMKII/CaMKII) and NADPH oxidase 4 (NOX(4)) were determined by Western blot analysis. The results showed that ISO-treated (i.p., daily 3 mg/kg, 2 weeks) rats developed an obvious cardiac hypertrophy as expressed by increases of HW/BW and myocyte cross-section area. Cardiac mitochondrial ROS level was significantly enhanced in ISO group as compared to CTRL group (P < 0.05). The expressions of NOX(4) and p-CaMKII in ISO group were also up-regulated as compared to CTRL group (1.4 and 1.6 times of CTRL, respectively, P < 0.05). NAC supplement significantly suppressed the hypertrophic development of heart in ISO-stimulated rats. The cardiac mitochondrial ROS level showed a significant decrease in rats of ISO+NAC group (P < 0.05 vs ISO). In accordance with this, ISO+NAC group rats also showed marked reductions in the expressions of NOX(4) and p-CaMKII/CaMKII compared to ISO group rats (P < 0.05). There were no significant differences of the detected indices between the rats from CTRL+NAC and CTRL groups. SBP showed no differences among four groups. These results suggest that both oxidative stress and CaMKII play important roles in sustained βAR-stimulated cardiac hypertrophy. NAC may suppress ISO-induced cardiac hypertrophy by down-regulating the expression of activated-CaMKII, and by reducing the level of oxidative stress originated from mitochondria and NADPH oxidase pathways.
Subject(s)
Animals , Male , Rats , Acetylcysteine , Pharmacology , Calcium-Calmodulin-Dependent Protein Kinase Type 2 , Metabolism , Cardiomegaly , Isoproterenol , Pharmacology , Mitochondria, Heart , Metabolism , Myocardium , Pathology , NADPH Oxidase 4 , NADPH Oxidases , Metabolism , Oxidative Stress , Rats, Wistar , Reactive Oxygen Species , Metabolism , Receptors, Adrenergic, beta , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the expressions of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the airway of bronchiectasis (BE) patients, and evaluate the effect of pseudomonas aeruginosa (PAE) on the expression of TIMP-1 and MMP-9.</p><p><b>METHODS</b>In this case-control study, subjects were divided into BE group and control group, and the BE group was further divided into PAE group and other bacteria group based on the culture results of bronchoalveolar lavage fluid (BALF). BALF was obtained by bronchoscopy, the expressions of MMP-9 and TIMP-1 were determined by ELISA, then the ratio of TIMP-1/MMP-9 was calculated. Furthermore, the tissue of bronchic endomembrane was obtained by transbronchial biopsy and the expressions of MMP-9 and TIMP-1 were determined using immunohistochemical method.</p><p><b>RESULTS</b>The levels of MMP-9 in the BALF of PAE group and other bacteria group were significantly higher than that in control group (P=0.0000 both), and the expressions of MMP-9 in bronchic endomembrane of PAE group and other bacteria group were also significantly higher (P=0.0421 and 0.0003, respectively). The level of TIMP-1 in BALF of PAE group was significantly lower than that in other bacteria group (P=0.0324). The ratio of TIMP-1/MMP-9 in BALF of BE group was significantly lower than that in control group(P=0.0000), and this ratio of PAE group was significantly lower than those in both other bacteria group and control group (P=0.0026 and 0.0000, respectively).</p><p><b>CONCLUSION</b>PAE infection in BE patients can suppress the expression of TIMP-1 and stimulate the expression of MMP-9, and thus make the disease even worse.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Bronchi , Metabolism , Microbiology , Bronchiectasis , Metabolism , Bronchoalveolar Lavage Fluid , Chemistry , Case-Control Studies , Matrix Metalloproteinase 9 , Metabolism , Pseudomonas aeruginosa , Tissue Inhibitor of Metalloproteinase-1 , MetabolismABSTRACT
<p><b>AIM</b>To explore the effects of sleep deprivation (SD) on myocardium and its antioxygen index.</p><p><b>METHODS</b>35 Sprague Dawley rats were randomly divided into five groups: Cage control, Tank control, SD 2 d, SD 4 d and SD 6 d. The "flower pot" technique was used to establish rats sleep deprivation model followed by record of surface electrocardiogram, detection of myocardium morphology changes under microscope and transmission electron microscope and investigation of MDA content and SOD activity of myocardial mitochondria.</p><p><b>RESULTS</b>After sleep deprivation, heart rate increased and ECG showed ischemia of myocardium; subcellular organelles such as chromosome, endoplasmic reticulum, mitochondria, intercalated disk impaired, myofibril lysis or necrosis, and lipid peroxidation reaction effects spread widely; edema, bleeding of the microvessels and invasion of the monocytes could be seen in the lumen. The MDA level increased and SOD activity increased followed by a decreased trend.</p><p><b>CONCLUSION</b>Sleep deprivation can induce damage on myocardium, and the stress especially oxygen stress caused by SD may be the possible mechanism.</p>
Subject(s)
Animals , Female , Male , Rats , Electrocardiography , Myocardial Ischemia , Pathology , Oxidative Stress , Physiology , Rats, Sprague-Dawley , Sleep Deprivation , Superoxide Dismutase , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To observe the clinical effect of Tiaoshen Tonglin Decoction (TTD)) on chronic prostatitis syndrome (CPS) and its effects on urinary flow rate (UFR), uric acid (UA) content and pH value in expressed prostate secretions (EPS).</p><p><b>METHODS</b>One hundred and eight patients with CPS were randomly assigned to two groups, the treatment group (56 cases) treated with TTI) and the control group (52 cases) with terazosin tablet, both for 60 days. The changes of chronic prostatitis symptom index (NIH-CPSL), established by the National Institutes of Health, U.S.A., UA and pH in EPS, as well as UFR were observed before and after treatment.</p><p><b>RESULTS</b>The cure rate and the total effective rate in the treatment group were higher than those in the control group respectively (P < 0.05); after treatment, the UA level, pH value in EPS and NIH-CPSI decreased significantly in the treatment group (P < 0.01), lower than those in the control group, which had significant change (P < 0.05); the maximum UFR and average UFR of both groups were improved markedly after treatment (P < 0.05) with insignificant difference between the groups.</p><p><b>CONCLUSION</b>TTD can improve the UFR, decrease the NIH-CPSI score, pH value and UA level in the EPS, is an effective recipe for treatment of CPS.</p>
Subject(s)
Adult , Humans , Male , Chronic Disease , Drugs, Chinese Herbal , Therapeutic Uses , Hydrogen-Ion Concentration , Pelvic Pain , Phytotherapy , Prostatitis , Classification , Drug Therapy , Syndrome , Treatment Outcome , Uric Acid , Chemistry , Metabolism , UrinationABSTRACT
Objective: To compare JGW with autogenous bone when used as grafting material in maxillary sinus floor augmentation.Methods: A study was conducted by performing a bilateral maxillary sinus floor augmentation with autogenous bone and JGW(Jin Gu Wei,Golden Bone,bone substitute,Shanghai Xiaobo Tec.) on rabbits.The process of bone formation was evaluated during different periods by imageology and histomorphology methods.Results: At 2nd week the gray scale of JGW was very lower than that of autogenous bone,but no significant difference was found at 12th week.The number of osteoblasts decreased with the time,no significant difference was found at 12th week.The degradation of JGW was relatively faster.Conclusion: JGW is feasible in maxillary sinus floor augmentation.