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To observe whether the mechanism of small dose capsaicin (Cap) against pulmonary fibrosis in mouse is mediated by agitating transient receptor potential vanilloid 1 (TRPV1). Methods: A total of 60 BALB/c mice were randomly divided into control (CON) group, bleomycin (BLM)group, Cap (0.5, 1,2 mg/kg) groups and Cap (2 mg/kg) plus SB-452533 (2.5 mg/kg) group. C57BL/6 mice were intratracheally injected with 3.5 mg/kg BLM to induce pulmonary fibrosis model. Animals for drugs treatment received daily drug via subcutaneous injection for 21 days. The morphological changes and collagen deposition in lung tissues were analysed by HE staining, Masson staining and immunohistochemistry. The concentration of calcitonin gene-related peptide (CGRP) in plasma was determined by ELISA. The mRNA and (or) proteins levels of α-CGRP, β-CGRP, collagen I, collagen III, E-Cadherin, zonula occludens-1 (ZO-1), vimentin, alpha smooth muscle actin (α-SMA), TRPV1, p-ERK1/2 and eukaryotic initiation factor 3a (eIF3a) were detected by qPCR and (or) Western blot. Compared with the BLM group, small dose Cap significantly reduced bleomycin-induced pulmonary fibrosis in mice and obviously reversed alveolar epithelial cells epithelial-mesenchymal transition (EMT) (the expression of E-cadherin and ZO-1 were increased(P＜0.05 or P＜0.01)and the expression of α-SMA and Vimentin were decreased (P＜0.05 or P＜0.01) after drugs treatment for 21 day, concomitantly with the increase the expressions of TRPV1 and CGRP (P＜0.05 or P＜0.01), and inhibiting ERK1/2 phosphorylation and eIF3a expression (P＜0.05 or P＜0.01). These effects of small dose Cap were abolished in the presence of TRPV1 receptor antagonist SB-452533. The results suggest that small dose Cap can reverse alveolar epithelial cells EMT and alleviate bleomycin-induced pulmonary fibrosis in mice by inhibiting ERK1/2/eIF3asignaling pathway, which is related to agitating TRPV1 receptor and releasing of CGRP.
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Objective@#To develop RT-nPCR assays for amplifying partial and complete VP1 genes of human enteroviruses (HEVs) from clinical samples and to contribute to etiological surveillance of HEV-related diseases.@*Methods@#A panel of RT-nPCR assays, consisting of published combined primer pairs for VP1 genes of HEV A-C and in-house designed primers for HEV-D, was established in this study. The sensitivity of each RT-nPCR assay was evaluated with serially diluted virus stocks of five serotypes expressed as CCID @*Results@#The sensitivity of RT-nPCR assays for amplifying partial VP1 gene of HEVs was 0.1 CCID @*Conclusion@#This RT-nPCR system is capable of amplifying the partial and complete VP1 gene of HEV A-D, providing rapid, sensitive, and reliable options for molecular typing and molecular epidemiology of HEVs in clinical specimens.
Subject(s)
Humans , Capsid Proteins/genetics , Enterovirus A, Human/genetics , Enterovirus B, Human/genetics , Enterovirus C, Human/genetics , Enterovirus D, Human/genetics , Molecular Epidemiology/methods , Molecular Typing/methods , Reverse Transcriptase Polymerase Chain Reaction/methodsABSTRACT
OBJECTIVE To explore the inhibitory effects of epalrestat (EPS) on platelet-derived growth factor (PDGF)-induced rat pulmonary artery smooth muscle cells proliferation by inhibiting aldose reductase (AR) expression.METHODS Primary rat pulmonary arterial smooth muscle cells (PASMCs) were prepared from the pulmonary artery of male 10-week-old Sprague-Dawley rats using explant method.PDGF 30 mg·L-1was given to induce cell proliferation.After PASMCs grew to 70%-80% conflu?ence, AR small-interferring RNA(ARsiRNA) was transfected with Lipofectamine 3000 into PASMCs. After 24 h,the expression and activity of AR were detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR),Western blotting and spectrophotometric method,respectively to investigate EPS on PASMCs proliferation and proliferating cell nuclear antigen (PCNA) and collagenⅠexpression induced by PDGF from in vitro. PASMCs (normal control, PDGF 30 mg·L-1, PDGF+EPS 1, 10 and 100 μmol·L-1,EPS 100 μmol·L-1)were treated according to groups.Cell proliferation was measured by BrdU marking and flow cytometry. The expressions of AR, PCNA and collagenⅠwere analyzed with RT-qPCR and Western blotting.RESULTS In cultured PASMCs,compared with normal control group, the application of exogenous PDGF-induced cell proliferation concomitantly up-regulated AR expres?sion and activity (P<0.01), and such effect was abolished by ARsiRNA. Compared with PDGF group, EPS attenuated PDGF-induced proliferation of PASMCs,expression of PCNA,and collagenⅠ(P<0.05, P<0.01),and the inhibitory effect of EPS was accompanied by inhibition of AR expression(P<0.05,P<0.01).CONCLUSION EPS inhibits PDGF-induced proliferation of PASMCs via inhibiting AR expression.
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A series of ursolic acid derivatives were synthesized by the introduction of 4-chloroindole compounds at A ring and esterification and amidation at C-28 position, which structures were characterized by 1H NMR, MS and etc. The cytotoxic activity of derivatives was evaluated against HepG2 and SGC7901 cells in vitro by MTT assay, in which paclitaxel and adriamycin were used as a positive control. The results indicated that all of derivatives can inhibit cell proliferation in HepG2 and SGC7901 cells with a better activity than ursolic acid. Especially, the compounds 6 and 12 showed significant antitumor activity comparable to the paclitaxel. The compounds are worthy to be studied further.
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Objective To investigate the level of homocysteine among healthy examination people and the possible related factors.Methods Retrospective analysis was performed to collect 1 259 results of healthy examination people from July to September in 2015,and 564 patients were confirmed to be hyperhomocysteinemia.The serum level of lipoids,glucose,uric acid and blood routine results were also collected.Results The incidence of hyperhomocysteinemia was 45.52％,and man has a higher rate than woman.The results of logistic regression showed positive results of UA (OR =1.006,95％ CI =1.005-1.008),HBG(OR =1.035,95％CI=1.026-1.045),and PLT (OR =0.996,95％ CI =0.993-0.998) in high hyperhomocysteinemia patients.Condusion High UA、HBG and low PLT levels are risk factors in hyperhomocysteinemia,and could be the important way for the early diagnoses of hyperhomocysteinemia.
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Objective:To analyze the role of acetylsalicylic acid (ASA) in immunomodulation of mesenchymal stem cells derived from gingiva (GMSCs),and to explore the role of ASA in enhancing the immumomodulation of GMSCs and the capacity of GMSCs to treat immune disorders and the underlying mechanism.Methods:Flow cytometry analysis were used to analyze the role of ASA in the expression of stem cells surface markers CD146,CD105,CD90,CD34 and CD45 in GMSCs,and the GMSCs proliferation was analyzed by 5-bromo-2-deoxyuridine (BrdU) staining and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.The GMSCs and T cells co-culture system was established to analyze the role of ASA in immunomodulation of GMSCs by measuring T cell apoptosis using flow cytometry analysis and inflammatory cytokines using enzyme linked immunosorbent assay (ELISA).Further more,the dextran sulfate sodium (DSS) induced colitis mouse model was established and the mouse body weight,disease activity score,histological index and pathological change of colons were analyzed after GMSC infusion.Results:The proliferation of GMSCs and the expressions of CD105,CD146 in GMSCs were increased after ASA treatment.In the GMSCs and T cells co-culture system,GMSCs induced T cells apoptosis and inhibited interferon γ (IFN-γ) and tumor necrosis factor α (TNF-α) secretion by T cells,which were enhanced by ASA treatment.In vivo,GMSCs infusion could ameliorate DSS-induced colitis,including inhibited DSS-induced mouse body weight loss,decreased disease activity score and histological index,and decreased inflammation cells infiltration in colons,as shown by hematoxylin-eosin (HE) staining.Moreover,the therapeutic effects of GMSC infusion on DSS-induced colitis could be enhanced by ASA treatment.Mechanically,ASA treatment increased FasL expression of Fas/ FasL death pathway in GMSCs to induce T cells apoptosis.Conclusion:ASA enhanced immunomodulation of GMSCs and increased the capacity of GMSCs to ameliorate DSS-induced colitis in mice.
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The experiment is designed to explore pathological festures and material basis of pseadoanaphylactoid reaction induced by notoginseng total saponin preparation. Mouse pseadoanaphylactoid reaction was used, 50 ICR mice were randomly assigned to control group, positive medicine group, notoginseng total saponin preparation low-dose group, notoginseng total saponin preparation middle-dose group, notoginseng total saponin preparation high-dose group on average. They are treated by intravenous injection of test substance solutions containing 0.4% Evans blue (EB). 30 min later, scores of ear blue staining and quantitation of ear EB exudation were recorded. Another two experiment were repeated in the same way excluding EB, just to. detect the related cytokines in serum using ELISA. We found that the scores of pseudoanaphylactoid reaction in notoginseng total saponin preparation injection middle-dose group and high-dose group was evidently higher than that in control group, suggesting that notoginseng total saponin preparation injection may be can lead to pseadoanaphylactoid reaction. HE staining showed that pseadoanaphylactoid reaction induced by notoginseng total saponin preparation injection is related to inflammation. Histamine, VEGF and TNF-α levels in notoginseng total saponin preparation middle-dose group and high-dose group significantly increased (P < 0.05, P < 0.01) than control group and showed a dose-dependent manner as well as consistent with the degree of ear blue dye. While IL-6 and IL-10 content did not increase significantly in notoginseng total saponin preparation low-dose group and middle-dose group, but they significantly higher than control group (P < 0.05, P < 0.01) when it increased to quadrupe clinical concentrations, eight times of the clinical dose. So pseadoanaphylactoid reaction caused by notoginseng total saponin preparation may be related to histamine, VEGF, TNF-α, and it is possible that IL-6 and IL-10 can play a role when pseadoanaphylactoid reaction achieve a certain high degree.
Subject(s)
Animals , Mice , Anaphylaxis , Capillary Permeability , Cytokines , Blood , Dose-Response Relationship, Drug , Drug Hypersensitivity , Mice, Inbred ICR , Panax notoginseng , Chemistry , SaponinsABSTRACT
<p><b>OBJECTIVE</b>To explore the relationship between aggressive behaviors, parent-child separation and experience of childhood abuse among junior high school students.</p><p><b>METHODS</b>A total of 1417 students in ordinary junior high schools from 3 townships in Huoshan, Anhui were involved in this study. Self-made questionnaire was used to estimate aggressive behaviors, parent-child separation in childhood, child abuse and social demographic information of the students under this study.</p><p><b>RESULTS</b>Related scores (2.52 ± 0.78) on physical aggression in boys was higher than in girls (2.29 ± 0.79) while the scores related to anger (2.60 ± 0.82) and hostility (2.58 ± 0.80) in girls, were higher than those in boys (2.41 ± 0.75, 2.47 ± 0.78), all with statistically significant differences (P < 0.05). Scores related to different types of aggressive behaviors and the scores in total, were higher in students from the senior class (P < 0.001). Scores on items as verbal aggression, hostility and in total, were higher in those adolescents which had undergone maternal-child separation during their childhood (P < 0.05). Scores on hostility and in total, were higher in those adolescents which had suffered from father-child separation during their childhood (P < 0.05). Scores related to anger, hostility and in total, were higher in those adolescents which had undergone both parent-child separation when they were much younger (P < 0.05). Students who had suffered from various types of repeated abuse showed higher scores in various types of aggressive behaviors and in total, than those who did not have the same experience. Most of the differences among groups were statistically significant (P < 0.05).</p><p><b>CONCLUSION</b>Students that suffered parent-child separation in their earlier childhood and with repeated experiences of abuse in childhood appeared to be risk factors causing aggressive behaviors to develop during the age of adolescence.</p>
Subject(s)
Adolescent , Child , Female , Humans , Male , Aggression , Child Abuse , Risk Factors , Schools , Students , Surveys and QuestionnairesABSTRACT
<p><b>OBJECTIVE</b>Erythropoietin and erythropoietin receptor (EPO-R) are expressed in many kinds of tumors. The EPO/EPO-R signaling is involved in tumor cell proliferation, invasion and angiogenesis. The aim of this study was to detect the expression of EPO-R in non-small cell lung cancer (NSCLC), and explore its correlation with angiogenesis.</p><p><b>METHODS</b>The expression patterns of EPO and EPO-R in 31 cases of NSCLC tissues were detected by immunohistochemistry, and that in benign lung lesions of 21 patients as control. To analyze the correlation of EPO/EPO-R expression patterns and clinicopathological factors. CD34 was used to label the vascular endothelial cells and calculate the microvessel density (MVD).</p><p><b>RESULTS</b>The positive rates of EPO and EPO-R expression in NSCLC were 67.7% and 96.8%, respectively, significantly higher than those in the control ones. The positive rates of EPO and EPO-R expression in adjacent tissues were 19.4% and 35.5%, and in benign lesions were 9.5% and 19.0%, respectively (P < 0.001). The expression patterns of EPO/EPO-R were not related with pTNM stage, histological type, histological grade and lymph node metastasis (P > 0.05). Increased MVD was correlated with poor differentiation, lymph node metastasis, and advanced stage.</p><p><b>CONCLUSIONS</b>High expression of EPO/EPO-R in NSCLC patients suggest that they may be involved in tumorigenesis. EPO/EPO-R expression and MVD are closely related, and they might be an endogenous stimulant of angiogenesis during the progression of non-small cell lung cancer. It may provide evidence for clinical diagnosis.</p>
Subject(s)
Humans , Antigens, CD34 , Metabolism , Carcinoma, Non-Small-Cell Lung , Metabolism , Pathology , Erythropoietin , Metabolism , Lung Neoplasms , Metabolism , Pathology , Lymphatic Metastasis , Microvessels , Pathology , Neoplasm Staging , Neovascularization, Pathologic , Receptors, Erythropoietin , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of compound granule prescription of thunberg fritillary bulb in relieving the post-chemotherapy bone marrow depression in refractory acute leukemia (RAL) patients.</p><p><b>METHOD</b>Two hundred and thirty eight RAL patients collected from 7 third-grade class-A hospitals were randomly divided into the treatment group and the control group. Both groups were treated with conventional chemotherapy. They were administered with compound granule prescription of thunberg fritillary bulb or placebo three days before chemotherapy for consecutively 14 days. A standardized chemotherapy course was a treatment cycle. The changes in peripheral hemogram of two groups were detected before and after chemotherapy.</p><p><b>RESULT</b>After the treatment, the white blood cell counts of the two groups were significantly different (P < 0.05), but there was no statistical significance in qualitative comparison. Before and after the treatment, the difference of the white cell counts of the two groups detected had significant statistics (P < 0.05). The white cell counts of both groups declined after chemotherapy, but the treatment group decreased more significantly than the control group. Before and after the treatment, there were no statistical significances in quantitative and qualitative comparisons both in HGB and PLT. After the treatment, HGB and PLT contents of both groups declined, but there was no statistical difference between the two groups.</p><p><b>CONCLUSION</b>Compound granule prescription of thunberg fritillary bulb can relieve the bone marrow suppression in RAL patients caused by the chemotherapy, which is mainly reflected by the slowdown of reduction in white blood cells.</p>
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Female , Humans , Male , Middle Aged , Young Adult , Acute Disease , Therapeutics , Antineoplastic Agents , Blood Cell Count , Bone Marrow , Drug-Related Side Effects and Adverse Reactions , Drugs, Chinese Herbal , Fritillaria , Chemistry , Leukemia , Blood , Drug TherapyABSTRACT
<p><b>OBJECTIVE</b>To investigate the molecular mechanism of haemophilia B caused by the novel mutation of Arg327Ile (R327I) in FIX gene.</p><p><b>METHODS</b>The R327I, R327Ala(A), R327Lys(K), R327Asn(N) and a replacement mutant (FIXβFVII), in which FIX β strand 324-329 was replaced by that of FVII 298-303, expression plasmids were constructed with site-directed mutagenesis method based on the wild-type (WT) FIX expression plasmid. The HEK293 cell was transiently transfected, then the activity of FIX (FIX:C) was assayed by one stage method in the conditioned medium, while the FIX:Ag in both the conditioned media and the cell lysates was measured by ELISA. The molecular weight and the semi-quantity of expressed FIX were analyzed by Western blot. Fluorescent protein expression plasmid was constructed to investigate the synthesis and secretion of the FIX R327I mutation in the viable cells.</p><p><b>RESULTS</b>FIX:C of the R327I mutant protein was 4.49% of the level of the WT in the conditioned medium, and the FIX:Ag of the R327I mutant protein in the conditioned medium and the cell lysates was 31.02% and 129.29% compared to that of WT, respectively. The mutation was characterized as cross-reaction material reduced (CRMR). The viable cell fluorescent assays showed that the R327I protein was more in both the viable cells and in lysosome than that of WT. The FIX:C of the R327A, R327K, R327N and FIXβFVII mutants was reduced compared to that of WT, the reduction of FIX:C of FIXβFVII was the most significantly amount among all the mutants in medium. FIX:Ag of all the mutants in the medium, except that the R327K increased, was reduced. The result of Western blot showed that the molecular weight of R327I protein was the same as that of WT, but the amount of the protein was much less compared with WT in the conditioned medium.</p><p><b>CONCLUSION</b>The abnormal synthesis and secretion as well as the abnormal function of the R327I mutant protein causes haemophilia B. The residue of R327 as well as the β strand domain of R327 located play important roles of the specific function of FIX.</p>
Subject(s)
Humans , Factor IX , Genetics , HEK293 Cells , Hemophilia B , Genetics , Pathology , Mutagenesis, Site-Directed , Mutation , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To find the major risk factors associated with gastric cardia cancer.</p><p><b>METHODS</b>We selected five high incidence areas of esophageal cancer and gastric cancer which have cancer registration system, i.e. Cixian and Shexian of Hebei Province, Linxian of Henan Province, Feicheng of Shandong Province and Zhuanghe of Liaoning Province. Fifty newly diagnosed cases of cardiac cancer after January 1, 2008 were selected from each cancer registration database. A uniform questionnaire, which was fully consulted by experts, was used. Population-based 1:3 case-control study was conducted in those areas. The study recruited 250 cases of cardiac cancer and 750 matched controls, which were investigated with the uniform questionnaire. The data were statistically analyzed by fitting-conditional Logistic analysis.</p><p><b>RESULTS</b>Smoking, passive smoking, alcohol drinking, irregular meal, improper dining posture, heavy taste, dried food, pickled food, fried food, hot food, gastrointestinal history, gastroesophageal reflux disease (GERD) can increase the risk of cardiac cancer. To eat more bean and high BMI are protective factors of the single factor logistic analysis. Gastrointestinal history (OR = 42.899), dried food (OR = 5.932), irregular meal (OR = 4.911), hot food (OR = 4.144), pickled food (OR = 3.287), passive smoking (OR = 2.355), and GERD (OR = 1.930) can increase the risk of cardiac cancer, eat more bean (OR = 0.254) and BMI ≥ 25 (OR = 0.492) are protective factors of the mixture factors logistic analysis.</p><p><b>CONCLUSIONS</b>Gastric cardia cancer is caused by environmental risk factors and genetic factors. Health education in high cardiac cancer incidence areas and primary prevention popularized into people's daily life will be beneficial to decreasing the incidence of gastric cardia cancer.</p>