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Objective:To compare the effects on DNA strand break induced by ultra-high dose rate (FLASH) electron beam and conventional irradiation, and investigate whether FLASH effect was correlated with a reduction of radiation response.Methods:Aqueous pBR322 plasmid was treated with FLASH (125 Gy/s) and conventional irradiation (0.05 Gy/s) under physioxia (4% O 2) and normoxia (21% O 2). Open circle DNA and linear DNA were detected by agarose gel electrophoresis, and the plasmid DNA damage was quantified with an established mathematical model to calculate the relative biological effect (RBE) of DNA damage. In some experiments, Samwirin A (SW) was applied to scavenge free radicals generated by ionizing radiation. Results:Under physioxia, the yields of DNA strand breakage induced by both FLASH and conventional irradiation had a dose-dependent manner. FLASH irradiation could significantly decrease radiation-induced linear DNA compared with conventional irradiation ( t=5.28, 5.79, 7.01, 7.66, P<0.05). However, when the aqueous plasmid was pretreated with SW, there was no difference of DNA strand breakage between FLASH and conventional irradiation ( P>0.05). Both of the yields of open circle DNA and linear DNA had no difference caused by FLASH and conventional radiotherapy at normoxia, but were significantly higher than those under physioxia. In addition, the yields of linear DNA and open circle DNA induced by FLASH irradiation per Gy were (2.78±0.03) and (1.85±0.17) times higher than those of conventional irradiation, respectively. Conclusions:FLASH irradiation attenuated radiation-induced DNA damage since a low production yield of free radical in comparison with conventional irradiation, and hence the FLASH effect was correlated with oxygen content.
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Objective:To analyze the data of ultra-high dose rate (FLASH) radiotherapy in GEO (Gene Expression Omnibus) database by bioinformatics method, in order to find the hub genes involved in flash radiotherapy induced acute T-lymphoblastic leukemia.Methods:The gene expression profiles of malignant tumors receiving FLASH radiotherapy were downloaded from GEO database. The R software was used to screen the differential expressed genes (DEGs) and analyze their biological functions and signal pathways. The protein-protein interaction (PPI) network of DEGs was analyzed by online tool of STRING, and Hub genes were screened by Cytoscape plug-in. The expressions of screened Hub genes in acute T lymphoblastic leukemia were identified with TCGA (The Cancer Genome Atlas) and GTEx (Genotype-Tissue Expression) database.Results:Based on the analysis of GSE100718 microarray dataset of GEO database, a total of 12 800 genes were found to be associated with radiosensitivity of acute T lymphoblastic leukemia, of which 61 significantly altered DEGs were selected for further analysis. It was found that these genes were involved in the biological processes of metabolism, stress response, and immune response through the pathways of oxidative phosphorylation, unfolded protein response, fatty acid metabolism, and so on. PPI analysis indicated that HSPA5 and SCD belonged to the Hub genes involved in the regulation of FLASH radiosensitivity, and they were significantly highly expressed in acute T lymphoblastic leukemia combined with TRD/LMO2-fusion gene.Conclusions:Through bioinformatics analysis, the Hub genes involved in regulating the sensitivity of FLASH radiotherapy and conventional radiotherapy can be effectively screened, and thus the gene expression profiles can be used to guide the stratification of cancer patients to achieve a precise radiotherapy.
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Objective:To investigate the clinical characteristics and therapeutic strategies of huge esophageal stones.Methods:One patient with esophageal esophageal calculus admitted to Beijing Friendship Hospital, Capital Medical University, who failed in undergoing endoscope treatment and shifted to an operation were retrospectively analyzed, and analyzed and summarized in combination with relevant literature.Results:The patient has significant dysphagia and severe malnutrition requiring further medical intervention. CT examination found that the lower esophagus was incarcerated with huge stones, and gastroscopy showed stones were obstructing the lumen, esophageal polyps and muscle bridges under the stones.Because the stones were large and hard, multiple endoscopic stone removal failed, and then the stones were successfully removed by thoracic surgery. The patient recovered well after surgery and had no complications such as anastomotic leakage or stenosis.Conclusions:Esophageal stones are easy to cause obstruction, and even lead to perforation, so it should be actively managed clinically. For patients with esophageal stones who have failed multiple endoscopic stone removals, thoracotomy can be used as a further treatment.
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Objective:To evaluate the efficacy and safety of stereotactic ablative radiotherapy (SABR) in patients with pulmonary oligometastases.Methods:Clinical data of patients with pulmonary metastases treated with SABR from 2011 to 2018 were retrospectively analyzed. The local control rate (LCR) and overall survival (OS) were calculated by Kaplan- Meier method. log-rank test was used for univariate analysis and Cox’s regression model for multivariate analysis. Results:A total of 214 lung metastases were detected in 159 patients, and the median follow-up time was 43 months. The 1-, 3-and 5-year LCR were 90.1%, 73.9% and 65.8%, respectively. The 1-, 3-and 5-year OS were 73.8%, 43.6% and 11.9%, respectively. Univariate analysis showed that biological effective dose (BED)≥100 Gy was significantly correlated with LCR ( P=0.033). Cox’s multivariate analysis showed that BED and primary tumor source were the independent prognostic factors of LCR ( P=0.023, P=0.043). No>grade 3 adverse events were observed in all patients during treatment. Conclusions:SABR is a safe and effective treatment of lung oligometastases. SABR should be actively aD ministered for pulmonary oligometastases, especially for those with lesions from lung cancer and the radiation dose should be selected as BED ≥100 Gy.
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Esophageal cancer is one of the common malignancies of the digestive tract, and its exact etiology and pathogenesis have not been completely clear. The establishment of animal model of esophageal cancer provides support for the basic and preclinical research of esophageal cancer, which is of great significance for further study to clarify the pathogenesis and explore the development of new therapeutic drugs. At present, the animal models commonly used in the study of esophageal squamous cell carcinoma mainly include chemical induction model, heterotopic transplantation model and orthotopic transplantation model. With the continuous progress of animal research on human origin tumor, the human transplanted tumor model has been more and more widely used. No matter which model has its own advantages and disadvantages and applicability, should be selected according to the purpose of the experiment. In this paper, the research progress on animal models of esophageal squamous cell carcinoma in recent years is reviewed, and the models commonly used in scientific research and preclinical treatment are discussed, which may provide a theoretical basis for the clinical treatment.
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Objective:To evaluate the long-term survival and identify prognostic factors of patients diagnosed with early-stage non-small cell lung cancer (ES-NSCLC) receiving stereotactic ablation radiotherapy (SABR).Methods:Clinical data of 109 ES-NSCLC patients treated with SABR in Henan Cancer Hospital from 2011 to 2018 were retrospectively analyzed. The overall survival (OS), cancer-specific survival (CSS) and progression-free survival (PFS) were calculated by Kaplan- Meier method and log-rank test. Multivariate prognostic analysis was performed by Cox regression model. Results:The median follow-up time was 44 months (2-93 months). The median OS, CSS and PFS were 78 months, 78 months and 44 months, respectively. The 1-year OS, CSS and PFS were 95.4%, 97.2% and 84.1%, and 75.6%, 79.1% and 56.6% for the 3-year OS, CSS and PFS, and 55.6%, 60.7% and 37.3% for the 5-year OS, CSS and PFS, respectively. Univariate analysis showed that ECOG score, age, smoking history and derived-neutrophil/lymphocyte ratio (dNLR) were the influencing factors of OS ( P=0.03, 0.02, 0.04, 0.001). Age, smoking history and dNLR were the influencing factors of CSS ( P=0.02, 0.03, 0.001). Multivariate analysis demonstrated that dNLR was an independent prognostic factor for OS and CSS ( P=0.001, 0.001). Conclusions:ES-NSCLC patients treated with SABR can achieve favorable survival. The dNLR is an independent prognostic factor of OS and CSS, which can be considered in clinical application.
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Objective:To investigate the effect of mannose on the radiosensitivity of six human non-small cell lung cancer cell lines and its possible mechanism.Methods:The expression of mannose phosphate isomerase in six lung cancer cell lines were detected by Western blot. The inhibitory effect of mannose on the proliferation of lung cancer cell lines were observed by MTT assay. When irradiated with 0, 2, 4, 6, 8 and 10 Gy, the effect of mannose on the radiosensitivity of six lung cancer cell lines was detected by plate clone formation assay, respectively; and the apoptosis rates of normal control, mannose, irradiation and combined groups were detected by flow cytometry.Results:The expression levels of mannose phosphate isomerase were different among six lung cancer cell lines. Among them, A549 cells had the highest expression level and H460 cells showed the lowest expression level. When aD ministrated with 11.1 mmol/L mannose, the same inhibitory effect was observed on both A549 and H460 cell lines. Moreover, the inhibitory effect on H460 cell line was significantly increased with the increase of mannose concentration. In addition, aD ministration of 11.1 mmol/L mannose could significantly increase the radiosensitivity and apoptosis rate of H460 cell line. However, it exerted limited effect upon the radiosensitivity and apoptosis rate of A549 cell line. Conclusion:In six lung cancer cell lines with high expression of mannose phosphate isomerase, the aD ministration of mannose can enhance the radiosensitivity of partial tumors cells.
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Objective:To evaluate the role of salvage radiotherapy in the re-treatment of patients with regional lymph node oligo-recurrence after radical surgery for esophageal squamous cell carcinoma.Methods:Clinical data of patients diagnosed with thoracic esophageal squamous cell carcinoma treated with radical surgery and developed regional lymph node oligo-recurrence ( n=1-3) from January 2013 to January 2016 were retrospectively analyzed. A total of 74 cases with intact clinical data were extracted for analysis. The survival analysis was performed by Kaplan-Meier method. Group comparison was conducted by Log-rank method. Results:The median overall survival (OS) after recurrence was 9(2.5-43) months, and the median progression-free survival time (PFS) was 4(1-33) months. There were 47 cases in the salvage radiotherapy group and 27 cases in the non-radiotherapy group, and the objective response rates were 77%(36/47) and 30%(8/27), respectively. Patients in the salvage radiotherapy group had better OS ( P=0.042) and PFS ( P=0.01) compared with their counterparts in the non-radiotherapy group. Among the patients who received salvage radiotherapy, involved field irradiation and elective nodal irradiation yielded similar OS ( P=0.963) and PFS ( P=0.599), and patients treated an irradiation dose ≥ 60Gy had better OS ( P=0.001) and PFS ( P=0.001) compared with those with dose< 60Gy. Conclusions:Local salvage radiotherapy is an effective treatment of esophageal squamous cell carcinoma with regional lymph node oligo-recurrence after radical surgery. Salvage radiotherapy has better OS and PFS compared with non-radiotherapy. Prospective clinical studies should be carried out to standardize the target and dose of radiotherapy, and to further clarify the effect of radiotherapy.
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Objective@#To compare the effect of neoadjuvant chemoradiotherapy (NCRT) and neoadjuvant chemotherapy (NCT) on the survival of patients with esophageal cancer.@*Methods@#Clinical data of 275 cases of thoracic esophageal squamous cell carcinoma treated with neoadjuvant therapy combined with surgery from December 2011 to December 2015 were analyzed retrospectively. The data of treatment and follow-up were complete and analyzable. There were 70 cases in the NCRT group and 205 cases in the NCT group. The survival rate was calculated by Kaplan-Meier method and statistically compared by log-rank test, and multivariate analysis was performed by Cox regression model.@*Results@#The median follow-up time was 32(3-84) months. The median survival time and recurrence-free survival time was 42(3-84) months and 30(3-84) months, respectively. The overall 3-and 5-year survival rates were 56.8% and 45.9%, respectively, and the 3-and 5-year recurrence-free survival rates were 45.1% and 38.9%, respectively. The median survival time in the NCRT and NCT groups was 46(7-84) and 40(4-74) months, and the median recurrence-free survival time was 31(3-84) and 28(3-69) months, respectively. The 3-and 5-year overall survival of the two groups were 59.1%, 47.1% and 56.3%, 47.5%(P=0.515), and the 3-and 5-year recurrence-free survival were 44.5%, 40.1% and 47%, 39%, respectively. There was no significant difference in the survival between two neoadjuvant therapy modes (P=0.554). Multivariate analysis showed that postoperative pathological TNM staging was an independent factor affecting the prognosis of patients with esophageal cancer (P=0.001).@*Conclusions@#The survival results of NCRT are similar to those of NCT. Postoperative pathological staging is an independent survival factor.
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OBJECTIVE： To study the relationship of CYP3A4， CYP2C8 and CYP3A5 gene polymorphism with ADR/blood concentration of hydroxychloroquine in patients with autoimmune disease （AID）， and to provide reference for individual medication of hydroxychloroquine. METHODS： Totally 77 AID patients，who were treated with hydroxychloroquine （daily dose of 200 mg to 400 mg） for a long-term （＞6 months）， were selected from the department of rheumatology and immunology， the First Affiliated Hospital of Anhui Medical University during Jul. 2017 to Aug. 2018. The information， blood sample and ADR of them were collected. Those patients were divided into normal liver function group， abnormal liver function group， normal renal function group， abnormal renal function group， normal eye group and abnormal eye group according to the site of ADR. The concentration of hydroxychloroquine was determined by HPLC. Genotype of CYP3A4， CYP2C8 and CYP3A5 were detected by MassARRAY microarray system. The differences of hydroxychloroquine-induced ADR in different genotypes were analyzed by χ2 test. The blood concentration difference of hydroxychloroquine in different genotypes were analyzed by independent sample t-test and one-way ANOVA. RESULTS： There was statistical significance in the distribution of CYP3A5 rs4646453 locus between normal renal function group and abnormal renal function group（P＜0.05）. The incidence of CC genotype was higher than that of AA+AC genotype in abnormal renal function group. There was statistical significance in the distribution of CYP2C8 rs10882526 locus between normal liver function group and abnormal liver function group（P＜0.05）. The incidence of allele G was higher than that of allele A in abnormal liver function group， and the incidence of AG genotype was higher than that of AA genotype. There was no significant correlation of the gene polymorphisms of CYP3A4， CYP2C8 and CYP3A5 with blood concentration among 77 AID patients. In subgroup analysis， blood concentration of GT， GG and TT genotypes of CYP2C8 rs10882521 in 58 patients with systemic lupus erythematosus （SLE） were 514.1，735.3 and 785.9 ng/mL， respectively； GG and TT genotypes were significantly higher than GT genotype （P＜0.05）. CONCLUSIONS： AID patients with CYP3A5 rs4646453 CC genotype have a higher incidence of renal dysfunction due to taking hydroxychloroquine； patients with CYP2C8 rs10882526 locus allele G and AG have a relatively high incidence of renal dysfunction due to taking hydroxychloroquine. When SLE patients taking the same dose of hydroxychloroquine， the blood concentration of hydroxychloroquine in patients carrying CYP2C8 rs10882521 GT genotype is lower than other genotypes.
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<p><b>OBJECTIVE</b>To investigate the antipyretic mechanism of Herba Ephedrae (Eph)-Ramulus Cinnamomi (RC) herb pair on yeast-induced pyrexia in rats.</p><p><b>METHODS</b>Totally 30 qualified male SD rats were randomly assigned to the normal control (NC) group, the pyrexia model (model) group, the Eph, RC and Eph-RC treatment groups by a random digital table, 6 rats in each group. Each rat received a 20% aqueous suspension of yeast (10 mL/kg) except the NC group. The 3 treatment groups were administered 8.1, 5.4 and 13.5 g/kg Eph, RC and Eph-RC respectively at 5 and 12 h after yeast injection, the NC group and the model groups were administered equal volume of distilled water. Rectal temperatures were measured at 0, 6, 8, 10, 12, 15, 18, 24 and 30 h and urine was collected prior to yeast injection and at 6, 10, 18, 24, 30, and 36 h after yeast injection. Then urine metabolomic profiling by gas chromatography tandem mass spectrometry, coupled with multivariate statistical analysis and pattern recognition techniques were used to explore the antipyretic effects of Eph-RC. Partial least squares discriminate analysis was used to analyze the metabolomics dataset including classification and regression in metabolomics plot profiling.</p><p><b>RESULTS</b>Compared with the NC group, rectal temperatures were significantly higher in the model group (P<0.01), while 3 treatment groups decreased significantly compared with the model group (P<0.05 or P<0.01). Rectal temperatures of Eph-RC-treated rats started to go down at 6 h, and markedly decreased at 8, 12, 15, 18 and 24 h (P<0.05 or P<0.01), while those of the Eph and RC groups had decreased firstly at 8 h and were markedly lower at 12 h (P<0.05 or P<0.01). Seventeen potential biomarkers related to pyrexia were confirmed and identified, including pyruvic acid, L-phenylalanine, L-tyrosine, phenylacetic acid, hippuric acid, succinic acid, citrate and so on. Eight potential alterations of metabolic pathways including phenylalanine metabolism, citrate cycle, tryptophan metabolism, biosynthesis of valine, leucine and isoleucine, were identified in relation to the antipyretic effects of Eph-RC using MetPA software.</p><p><b>CONCLUSION</b>The antipyretic effect of Eph-RC herb pair on yeast-induced pyrexia in rats involved correction of perturbed amino acid, fatty acid, and carbohydrate metabolism according to the metabolic pathway analysis with MetPA.</p>
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<p><b>OBJECTIVE</b>To assess the antinociceptive and anti-inflammatory properties of the aqueous extract of Armadillidium vulgare (AV).</p><p><b>METHODS</b>The antinociceptive effect of AV (400, 600 and 800 mg/kg) was investigated in mice using the acetic acid-induced writhing, formalin-induced nociceptive, and hot plate tests. Phlogogen-induced paw edema using carrageenan, dextran, or compound 48/80 as phlogogen was used as inflammatory models to evaluate AV's anti-inflammatory effect. Additionally, the bioactive substances glucosamine (GLcN) and taurine in AV were determined using high-performance liquid chromatography.</p><p><b>RESULTS</b>Oral treatment of the mice with AV (600 and 800 mg/kg) significantly reduced the number of writhes in the acetic acid-induced writhing test (P<0.01) but not the hot plate test (P>0.05). All doses tested significantly inhibited paw-withdrawal during the second phase of the formalin-induced nociceptive model (P<0.01). AV demonstrated a strong anti-inflammatory effect in all those inflammatory models (P<0.05).</p><p><b>CONCLUSIONS</b>AV has antinociceptive and anti-inflammatory effects, providing scientific evidence of the efficacy of its traditional use in pain treatment. Furthermore, GLcN and taurine contribute, at least in part, to the anti-inflammatory activity of AV.</p>
Subject(s)
Animals , Female , Male , Mice , Analgesics , Pharmacology , Anti-Inflammatory Agents , Pharmacology , Edema , Drug Therapy , Inflammation , Drug Therapy , Isopoda , Chemistry , Pain , Drug Therapy , Pain Measurement , Phytotherapy , Plant Extracts , Chemistry , Pharmacology , Water , ChemistryABSTRACT
OBJECTIVE: To investigate the health status of workers exposed to 7-aminocephalosporanic acid( 7-ACA) and6-aminopenicillanic acid( 6-APA) in an antibiotics enterprise. METHODS: Using simple random sampling method,207 workers exposed to 7-ACA and 6-APA from an antibiotic production enterprise were selected as the exposed group,and 162 workers with no dust exposure history from the same antibiotic production enterprise were selected as control group. Health examinations were performed. The health status of the workers were analyzed. RESULTS: The detection of symptoms( chest distress,shortness of breath,cough,wheezing,itchy skin) and allergic diseases( bronchial asthma,allergic rhinitis,allergic dermatitis) in exposed group were higher than those in control group( P < 0. 05). The lung function indexes such as forced vital capacity( FVC),forced expiratory volume in first second( FEV1) and FEV1/ FVC in the exposed group were lower than those in control group( P < 0. 05). There was no significant difference in the lung function indexes,respiratory and skin allergy symptoms,and allergic diseases between 7-ACA subgroup and 6-APA subgroup( P < 0. 017).The incidence of bronchial asthma,allergic rhinitis had statistical differences in the length of dust exposure in service workers( P < 0. 05). CONCLUSION: Workers exposed to 7-ACA and 6-APA have a high occurrence rate of respiratory symptoms. These workers also suffered from occupational diseases such as bronchial asthma,allergic rhinitis and others.
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<p><b>OBJECTIVE</b>Mahuang-Shigao herb-pair is a famous formula composed of Ephedra and Gypsum. The herb-pair is frequently used for treating cold symptoms and bronchial asthma in the clinical practice of Chinese medicine (CM). In the present study, we evaluated evidence for the benefit of combined use of Ephedra and Gypsum by analyzing the antipyretic and anti-asthmatic activities of Ephedra-Gypsum.</p><p><b>METHODS</b>The antipyretic effects of Ephedra-Gypsum were evaluated in yeast-induced hyperthermia test. Thirty male Wistar rats were randomly divided into 5 groups, including control group, standard aspirin group, and 3 Ephedra- Gypsum groups of different doses (6, 12, 24 g/kg). Ephedra-Gypsum extract and asprin were administered orally 6 h after the injection of yeast solution and body temperature was measured every 1 h for 8 h. The antiasthmatic effects of Ephedra-Gypsum were evaluated using an ovalbumin (OVA)-induced asthmatic rat model. Thirty-six male SD rats were randomly divided into 6 groups. Rats were alternately sensitized and OVA+Al(OH) challenged by exposure to mists of ovalbumin. Ephedra-Gypsum extracts (6, 12, 24 g/kg) or dexamethasone were administered 45 min prior to the allergen challenge for 8 days. Latent period and the weight of wet to dry ratio of lung were determined. In addition, the eosinophils in blood and white blood cell (WBC) were counted by an YZ-Hemavet Analyzer.</p><p><b>RESULTS</b>The Ephedra-Gypsum extracts at test dose (6, 12, 24 g/kg) significantly and dose-dependently attenuated yeast-induced fever in rats. The Ephedra-Gypsum extracts also prolonged the latent period, reduced OVA-induced increases in eosinophils and WBC, and decreased the wet and dry weight ratio of the lungs in the anti-asthmatic test.</p><p><b>CONCLUSIONS</b>These findings indicate that the Ephedra-Gypsum extract has antipyretic and anti-asthmatic properties. Hence, the results support additional scientific evidence in prescriptions.</p>
Subject(s)
Animals , Male , Alkaloids , Anti-Asthmatic Agents , Therapeutic Uses , Antipyretics , Therapeutic Uses , Asthma , Drug Therapy , Calcium Sulfate , Therapeutic Uses , Drugs, Chinese Herbal , Therapeutic Uses , Ephedra , Chemistry , Fever , Drug Therapy , Lung , Pathology , Organ Size , Ovalbumin , Plant Extracts , Therapeutic Uses , Rats, Sprague-Dawley , Rats, WistarABSTRACT
In this study, UPLC-MS/MS was adopted to determine the contents of five ephedrine alkaloids (Norephedrine, Norpseudoephedrine, Ephedrine, Pseudoephedrine, Methylephedrine) in plasma and urine in rats after the combined administration of Ephedrae Herba-Gypsum Fibrosum and calculate relevant pharmacokinetic parameters, in order to discuss the effect of the combined administration of Ephedrae Herba-Gypsum Fibrosum on plasma pharmacokinetics and urinary excretion characteristics. According to the results, after being combined with Gypsum, the five ephedrine alkaloids showed similar pharmacokinetic changes, such as shortened t(max), accelerated absorption rate, but reduced AUC(0-t) and V(z)/F, which may be related to the increase in urine excretion. Besides, Gypsum was added to enhance C(max) of Pseudoephedrine and prolong MRT(0-t) of Methylephedrine, so as to enhance the anti-asthmatic effect of Ephedrae Herba and resist the toxic effect of Norephedrine and Ephedrine. This study proved the scientific compatibility of Ephedrae Herba-Gypsum Fibrosum and provided a reference for studies on the prescription compatibility regularity and relevant practices.
Subject(s)
Animals , Male , Rats , Alkaloids , Blood , Pharmacokinetics , Urine , Calcium Sulfate , Pharmacokinetics , Drugs, Chinese Herbal , Pharmacokinetics , Ephedra , Chemistry , Rats, Sprague-Dawley , Urine , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To study and compare the anti-inflammatory effect and molecular mechanism of artemisinin and dihydroartemisinin.</p><p><b>METHOD</b>Mouse mononuclear macrophage RAW264.7 cells were stimulated to release inflammatory mediators such as TNF-alpha, IL-6 and NO, in order to assess the drugs' inhibitory effect on macrophage's release of above inflammatory mediators. The levels of TNF-alpha and IL-6 were determined by ELISA and the cytotoxicity was determined by MTT method. The protein expression of iNOS, COX-2 and beta-actin were tested by Western blot. The enzymatic activity of COX-2 was determined by colorimetric method.</p><p><b>RESULT</b>Dihydroartemisinin significantly inhibited LPS-induced release of TNF-alpha, IL-6 and NO from RAW264.7 in mice with the concentration range of 12.5 - 100 micromol x L(-1), and showed good dose dependence. Artemisinin only inhibited the IL-6 release to a certain extent.</p><p><b>CONCLUSION</b>Dihydroartemisinin inhibits macrophages from releasing inflammatory factors TNF-alpha and IL-6 and inflammatory mediators NO by down-regulating iNOS protein. Artemisinin may help dihydroartemisinin to show its anti-inflammatory effect through metabolism.</p>
Subject(s)
Animals , Mice , Anti-Inflammatory Agents , Pharmacology , Artemisinins , Pharmacology , Cell Line , Gene Expression , Inflammation Mediators , Allergy and Immunology , Interleukin-6 , Genetics , Allergy and Immunology , Macrophages , Allergy and Immunology , Nitric Oxide , Allergy and Immunology , Tumor Necrosis Factor-alpha , Genetics , Allergy and ImmunologyABSTRACT
In this study,the coding region of type O FMDV capsid protein VP1 and a series of codon optimized DNA sequences coding for VP1 amino acid residues 141-160(epitopel),tandem repeat 200-213(epitope2(+2))and the combination of two epitopes(epitope1-2)was genetically cloned into the prokaryotic expression vector pPROExHTb and pGEX4T-1,respectively.VP1 and the fused epitopes GST-E1,GST-E2(+2)and GST-E1-2 were successfully solubly expressed in the cytoplasm of Escherichia coli and Western blot analysis demonstrated they retained antigenicity.Indirect VP1-ELISA and epitope ELISAs were subsequently developed to screen a panel of 80 field pig sera using LPB-ELISA as a standard test.For VP1-ELISA and all the epitope ELISAs,there were clear distinctions between the FMDV-positive and the FMDV-negative samples.Cross-reactions with pig sera positive to the viruses of swine vesicular disease virus that produce clinically indistinguishable syndromes in pigs or guinea pig antisera to FMDV strains of type A,C and Asial did not occur.The relative sensitivity and specificity for the GST-E1 ELISA,GST-E2(+2),GST-E1-2 ELISA and VP1-ELISA in comparison with LPB-ELISA were 93.3% and 85.0%,95.0% and 90%,100% and 81.8%,96.6% and 80.9% respectively.This study shows the potential use of the aforementioned epitopes as alternatives to the complex antigens used in current detection for antibody to FMDV structural proteins.
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In order to develop an anti-FMDV Asial type monoclonal antibody (mAb), BABL/c mice were immunized with recombinant FMDV VP1 protein. Three mAbs, 1B8, 5E1 and 5E2, were then further optimized. The result indicated that prepared anti-FMDV Asial mAbs had no cross-reactivity with Swine vesicular disease (SVD) and FMDV O, A and C type antigen. Their titers in abdomen liquor were l:5×106, l:2×106 and l:5×l06, respectively. 1B8 was found to be of IgGi subtype, 5E1 and 5E2 belonged to IgG2b subtype. In this study, the prepared mAbs are specific for detecting FMDV type Asial, and is potentially useful for pen-side diagnosis.