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RATIONALE: To investigate T lymphoma invasion and metastasis inducing factor 2 (Tiam2) protein for expression in esophageal carcinoma and relationship with clinical features among cases with tumors. PATIENT CONCERNS: In primary esophageal cancer patients, surgical resection of tumor tissue was performed in 65 cases and adjacent normal esophageal tissue in 20 cases. DIAGNOSES: Primary esophageal carcinoma (57 cases squamous cell carcinoma, 8 cases adenosquamous carcinoma). INTERVENTIONS: The expression level of Tiam2 protein in esophageal carcinoma tissues and normal esophageal tissues by SP immunohistochemical method. The expression intensity was quantitatively analyzed by using Image-pro plus software for image analysis, while SPSS26.0 software was used for a statistical analysis on the data. OUTCOMES: Tiam2 was highly expressed in esophageal squamous cell carcinoma and adenosquamous cell carcinoma, but low expressed in normal esophageal tissue. The expression level of Tiam2 protein was not correlated with gender and age of patients (Pâ >â .05), but was correlated with lymph node metastasis of esophageal carcinoma, TNM stage and differentiation degree of esophageal squamous cell carcinoma (Pâ <â .05). Tiam2 was positively correlated with Tiam1 for protein expression in esophageal carcinoma (râ =â .704, Pâ <â .001). LESSONS: The increased expression of Tiam2 protein in esophageal cancer may be an early molecular event of esophageal cancer. Tiam2 protein has a high expression level in esophageal carcinoma related to lymph node metastasis, TNM stage and differentiation degree, which suggests that Tiam2 protein plays an important role in the invasion and metastasis of esophageal carcinoma. There is a positive correlation between Tiam2 and Tiam1 protein expressions in esophageal carcinoma, suggesting that the 2 proteins may have a definite internal relationship.
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Neoplasias Esofágicas , Humanos , Neoplasias Esofágicas/patología , Neoplasias Esofágicas/metabolismo , Masculino , Femenino , Persona de Mediana Edad , Anciano , Factores de Intercambio de Guanina Nucleótido/metabolismo , Metástasis Linfática , Carcinoma Adenoescamoso/patología , Carcinoma Adenoescamoso/metabolismo , Proteína 1 de Invasión e Inducción de Metástasis del Linfoma-T/metabolismo , Carcinoma de Células Escamosas de Esófago/metabolismo , Carcinoma de Células Escamosas de Esófago/patología , Biomarcadores de Tumor/metabolismo , Adulto , Estadificación de Neoplasias , Carcinoma de Células Escamosas/patología , Carcinoma de Células Escamosas/metabolismo , InmunohistoquímicaRESUMEN
Objective To explore the association between serum amyloid A (SAA) and carotid intima-media thickness (CIMT) in patients with type 2 diabetes mellitus(T2DM).Methods A total of 148 diabetic subjects were divided into three groups according to C1MT:normal IMT group,IMT incrassation group and arteriosclerosis group.Levels of SAA,25-hydroxyvitamin D [25 (OH) D],brachial-ankle artery pulse wave velocity (baPWV),high density lipoprotein cholesterol (HDL-C),low-density lipoprotein cholesterol (LDL-C),fasting plasma glucose (FPG),glycosylated hemoglobin (HbA1 c),triglyceride (TG),total cholesterol (TC),body mass index (BMI) and blood presure were measured in all groups.The relationship between SAA,CIMT,25 (OH)D,baPWV and other factors was also analyzed.Results Compared to the normal IMT group,levels of FPG,HbA1 c,TC,BMI,SAA and ba-PWV were significantly higher in IMT incrassation group and arteriosclerosis group,while HDL-C and 25 (OH)D were lower.In arteriosclerosis group,levels of BMI,FPG,HbA1 c,SAA and ba-PWV were higher than those in IMT incrassation group,while 25 (OH) D was lower.Pearson correlation analysis showed that the level of SAA was positively correlated with baPWV,BMI,TC,course of disease and IMT,while it was negatively correlated with HDL-C and 25 (OH)D.Logistic regression analysis of IMT showed that smoking,obesity,high levels of HbAlc,FPG,TC,SAA,low levels of HDL-C and 25 (OH)D may contribute to higher levels of IMT.Conclusions SAA was closely related to carotid atherosclerosis.Further prospective studies will be helpful to explore the influence of SAA on diabetic macroangiopathy.
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OBJECTIVE: To provide reference for the establishment of quality standard of Shenhuang liniment. METHODS: Qualitative identification of Sophora flavescens, Phellodendron chinense, Rhei Radix Et Rhizoma and Scutellaria baicalensis in Shenhuang liniment were carried out by TLC according to the method of 2015 edition of Chinese Pharmacopeia (part Ⅳ). HPLC method was used to determine the contents of matrine and oxymatrine in Shenhuang liniment [Phenomenex Luna NH2 column, mobile phase consisting of acetonitrile-absolute ethanol-3% phosphoric acid aqueous solution (80 ∶ 10 ∶ 10,V/V/V), column temperature of 45 ℃, flow rate of 1.0 mL/min, detection wavelength of 220 nm, sample size of 5 μL]. RESULTS: Results of TLC showed that the corresponding spots of the same color were found in the corresponding positions of the chromatograms for test samples and reference substance/substance control; and the spots were clear, retardation factor was moderate, the separation degree was high, and the negative control had no interference. Results of HPLC showed that the linear range of matrine and oxymatrine were 203.60-1 221.60 ng(r=0.999 4)and 210.08-840.30 ng(r=0.999 7), respectively. RSDs of precision, stability and reproducibility tests were all lower than 3.0% (n=6). Average recovery rates were 96.03% and 100.93%, and RSDs were 2.55% and 2.69%(n=9). CONCLUSIONS: Established method is specific, accurate and reproducible, and can be used for quality control of Shenhuang liniment.
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AIM To establish the HPLC fingerprints of Cajanus cajan (L.) Millsp.leaves and to determine the contents of orientoside and luteolin.METHODS The analysis of 65% methanol extract from C.cajan leaves was performed on a 25 ℃ thermostatic Agilent Zorbax SB-C18 column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of methanol-1% acetic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 260 nm.RESULTS There were twenty-one common peaks in ten batches of samples (S1-S10),whose similarities were more than 0.950,except for that of S3 (0.516).Orientoside and luteolin showed good linear relationships within the ranges of 0.089 5-3.960 μg and 0.015 5-0.408 μg,whose average recoveries were 99.43% (RSD =1.32%) and 98.50% (RSD =0.82%),respectively.The contents of two constituents in the samples from three growing areas (Guangdong,Yunnan and Hainan) showed obvious differences.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of C.cajan leaves.
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OBJECTIVE:To establish UPLC fingerprint of Gehua formula granules. METHODS:UPLC method were adopted. The determination was performed on Zorbax Eclipse XDB-C18 column with mobile phase consisted of acetonitrile-water at the flow rate of 0.5 mL/min. The detection wavelength was set at 264 nm,column temperature was 25 ℃,and the sample size was 1 μL. Using tectorigenin as reference substance,UPLC chromatograms of 10 batches of Gehua formula granules were determined. The common peak identification and similarity evaluation were conducted by TCM Chromatogram Fingerprint Similarity Evaluation Sys-tem(2004 A edition). RESULTS:14 common peaks were identified in UPLC chromatograms of 10 batches of Gehua formula gran-ules and similarities were all higher than 0.90. UPLC chromatograms of 10 batches of samples were in good agreement with control fingerprint. CONCLUSIONS:Established UPLC fingerprint can provide reference for identification and quality evaluation of Gehua formula granules.
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Mycoplasma pneumoniae is thought to be the simplest and smallest cell wall-deficient bacterium which can cause chronic respiratory infections. Recently vaccination has been a possible and reliable way to reduce the spreading and infection effectively. In this study, the transmembrane proteins P116N (the N-terminal of P116), P1C (the C-terminal of P1), P30, and P116N-P1C-P30 (MP559 for short), a chimeric protein were expressed using prokaryotic expression system. The four purified recombinant proteins were synergized with freund's adjuvant and immunized New Zealand White rabbits respectively for three times. The IgG antibodies collected from immunized rabbits and mouse were analyzed by ELISA to analyze the immunogenicity and antigenicity. The results showed the four different recombinant proteins could induce strong humoral immune response. Protein MP559 could react with antisera from rabbit immunized with P1C, P30, and P116N, indicating MP559 was well designed and presented antigen epitopes of all the three antigens. Antibodies against P116N, P1C, and P30 could be stimulated by MP559 immunization, indicating MP559 has a potential to replace the three antigens as a vaccine candidate. This study laid a substantial foundation for the vaccine development of M. pneumoniae, and at the same time provided a essential strategy for the vaccine research of other pathogen.
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Adhesinas Bacterianas/inmunología , Vacunas Bacterianas/inmunología , Mycoplasma pneumoniae/inmunología , Neumonía por Mycoplasma/prevención & control , Adhesinas Bacterianas/genética , Animales , Anticuerpos Antibacterianos/inmunología , Vacunas Bacterianas/genética , Clonación Molecular , Inmunidad Humoral , Inmunización , Inmunoglobulina G/inmunología , Mycoplasma pneumoniae/genética , Neumonía por Mycoplasma/inmunología , Conejos , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunologíaRESUMEN
Objective: To study the antibacterial activity in vitro of Sanhuang Shaoshangling on several common bacteria after changing the preparation process. Methods:Using the new technology of ethanol reflux extraction and water extraction, Sanhuang Sha-oshangling was prepared. A micro-broth dilution method was used to determine the minimum inhibitory concentration ( MIC) of San-huang Shaoshangling for escherichia coli, pseudomonas aeruginosa, staphylococcus aureus and type B paratyphoid salmonella after chan-ging the preparation process. The antibacterial effect of Sanhuang Shaoshangling prepared by the new and the old technology was com-pared, and the inhibition zone was detected by a paper strip method. Results:The antibacterial effect of Sanhuang Shaoshangling pre-pared by the new technology on escherichia coli, pseudomonas aeruginosa, staphylococcus aureus and type B paratyphoid salmonella was better than that of the products prepared by the old technology with the minimum inhibitory concentration ( MIC) of 16 μg·ml-1 , 16μg·ml-1 , 8 μg·ml-1 and 16 μg·ml-1 , respectively, while that of the product prepared by the old technology was 32 μg·ml-1 , 64 μg·ml-1 , 16 μg·ml-1 and 64 μg·ml-1 , respectively. Conclusion: The antibacterial effect of Sanhuang Shaoshangling after changing the preparation process is more significant, which shows certain clinical significance.
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Objective:To establish a method for the separation and determination of ketoprofen enantiomer .Methods:A pre-col-umn derivation RP-HPLC method was used with L-alanine-β-naphthylamine ( L-Ala-β-NA) as the derivation reagent .The RP-HPLC conditions were as follows: a Hypersil ODS-2 column (250 mm ×4.6 mm,5 μm) was applied, the mobile phase was acetonitrile-0.025 mol· L-1 phosphate buffer solution (40∶60, v/v) and the flow rate was 1.0 ml· min-1 , the detection wavelength was set at 245 nm and the column temperature was 25℃.The injection volume was 10μl.Results:Base line separation was achieved for the sep-aration of enantiomer from ketoprofen , and the retention time for S-(+) -ketoprofen and the R-(-) -ketoprofen was 24.2 min and 26.0 min, respectively.Dexketoprofen within the range of 0.025-0.125 mg had a good linear relationship (r=0.998 1) and the aver-age recovery was 90.93%(RSD =4.10%, n=9 ).Conclusion:The method is simple, accurate and reliable, which can be applied in the separation and determination of ketoprofen .
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Objective To investigate Schistosoma japonicum infection on mice high-fat diet of insulin resistance. Methods 36 male C57 BL/6 J mice were randomly assigned into three equal groups: normal control group ( NC group) , high-fat diet group ( HF group) and high-fat diet with Schistosoma japonicum infected group ( HSJ group) . Specimen was collected 6 and 12 weeks after high-fat diet, separately. The levels of fasting blood glucose ( FBG) , fasting plasma insulin resistance index ( FINS) and insulin ( HOMA-IR) were detected. Interferon-γ( IFN-γ) ,in-terleukin-4 ( IL-4 ) and singal transductor and activator of transcription-4 ( STAT4 ) singal transductor and activator of transcription-6 (STAT6)were detected by ELISA and immunohistochemical method. Results The mice from HF group showed higher levels of HOMA-IR than those from NC groups by the end of 6 and 12 weeks after infection( P<0. 01 );the levels of HOMA-IR in mice from HSJ group were lower than NC group and HF group by the end of 12 weeks(P<0. 05);the levels of IL-4 in mice from HSJ group were higher than NC group and HSJ group by the end of 6 and 12 weeks after infection ( P<0. 05 ); the levels of STAT6 in mice from HSJ group were higher than HF group by the end of 12 weeks after infection ( P<0. 05 );the levels of STAT4 in mice from HF group were higher than NC group by the end of 6 and 12 weeks after infection. Conclusion Schistosome japonicum chronic infection may improve insulin resistance in obese mice with induced STAT6 protein expressed in liver tissue and secrete IL-4,providing new ideas for the prevention and treatment of diabetes.
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Aim To study the inhibitory activities of potential new anti-influenza virus agents,3-O-β-chaco-triosyl pentacyclic triterpenoids against the entry of H5N1influenza viruses.Methods Three target com-pounds were designed and synthesized structurally re-lated to the lead compound 3-O-β-chacotriosyl dioscin derivative (1 )with inhibitory activities against H5N1 influenza viruses.The inhibitory activities of these tar-get compounds were tested at a cellular level pseudo vi-rus system targeting H5N1 influenza viruse entry.Re-sults All the compounds 1 a,1 b and 1 c showed po-tent inhibitory activities against the entry of A/Thai-land/Kan353/2004 pseudo virus into the target cells, of which compound 1 b showed the best inhibitory activ-ity with an IC50 value of (1.25 ±0.22)μmol·L-1. Conclusion The SARs analysis of these compounds indicated that replacement of the aglycone moiety of compound 1 with pentacyclic triterpenoids could in-crease antiviral activity.Different types of pentacyclic triterpen as aglycone residue had the significant influ-ence on the inhibitory activity (1 b >1 c >1 a),sug-gesting ursane type of triterpenes was superior to the two other kinds of triterpenes as aglycone residue.
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Objective To investigate the effects of Schistosoma japonicum infection on selective activation of macrophages and insulin resistance in liver tissues of mice with high-fat diet induced obesity and the possible mechanism. Methods Thirty-six male C57BL/ 6J mice were randomly divided into 3 groups in-cluding normal control group(NC group,n=12),high-fat diet feeding group(HF group,n=12)and high-fat diet feeding with Schistosoma japonicum infection group(HSj group,n=12). Specimens were collected 6 and 12 weeks after high-fat diet feeding. The levels of fasting blood glucose(FBG),fasting plasma insulin (FINS)and homeostasis model assessment for insulin resistance index(HOMA-IR)were detected. The ex-pression of interleukin-6(IL-6),arginase-1(Arg-1)and found in inflammatory zone-1(Fizz-1)at mRNA and protein levels were detected by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR)and immunohistochemistry,respectively. Results The mice from HF group showed higher levels of HOMA-IR than those form NC group and HSj group by the end of 6 and 12 weeks after infection(P﹤0. 05). The levels of HOMA-IR in mice from HF group were increased by the end of week 12 after infection as com-pared with those at week 6(P>0. 05). The levels of IL-6 in mice from both HF group and HSj group were higher than those from NC group by the end of week 6 after infection(P﹤0. 05). Higher levels of IL-6 were detected in mice from HF group as compared with those from HSj group and NC group by the end of week 12 after infection(P﹤0. 05). The expression of Arg-1 and Fizz-1 in mice form HF group were lower than those from NC group by the end of 6 and 12 weeks after infection(P﹤0. 05). Arg-1 was highly expressed in mice form HSj group,followed by those from NC and HF groups 12 weeks after infection. The expression of Fizz-1 in mice from HSj group was the highest among the three groups by the end of week 6 and 12 after infection (P﹤0. 05). Conclusion The proinflammatory effects on mice with diet induced obesity were induced dur-ing acute infection of Schistosoma japonicum cercariae(6 weeks). The chronic infection of Schistosoma ja-ponicum cercariae(12 weeks)might be helpful in reversing hepatic insulin resistance in mice with diet in-duced obesity by changing the polarity of macrophages in liver tissues.
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Marine Actinobacteria are emerging as new resources for bioactive natural products with promise in novel drug discovery. In recent years, the richness and diversity of marine Actinobacteria from the South China Sea and their ability in producing bioactive products have been investigated. The objective of this work is to isolate and identify bioactive secondary metabolites from a marine actinobacterium SCSIO 1934 derived from sediments of South China Sea. The strain was identified as a Streptomyces spieces by analyzing its 16S rDNA sequence. Streptomyces sp. SCSIO 1934 was fermented under optimized conditions and seven bioactive secondary metabolites were isolated and purified by chromatographic methods including colum chromatography over silica gel and Sephadex LH-20. Their structures were elucidated as 17-O-demethylgeldanamycin (1), lebstatin (2), 17-O-demethyllebstatin (3), nigericin (4), nigericin sodium salt (5), abierixin (6), respectively, by detailed NMR spectroscopic data (1H, 13C, COSY, HSQC and HMBC). This work provided a new marine actinobacterium Streptomyces sp. SCSIO 1934, capable of producing diverse bioactive natural products.
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Antibacterianos , Química , China , ADN Ribosómico , Química , Genética , Sedimentos Geológicos , Microbiología , Océanos y Mares , ARN Ribosómico 16S , Genética , Streptomyces , Química , Clasificación , GenéticaRESUMEN
<p><b>OBJECTIVE</b>To study the gene mutation and streptomycin, isoniazid or rifampicin resistance of Mycobacterium isolated from silico-tuberculosis patient's sputum so as to find a more effective therapy for this disease.</p><p><b>METHODS</b>Mycobacteria tuberculosis were separated from 96 coal worker with silico-tuberculosis firstly. Then rpsL, KatG and rpoB fragments of genome were copied with PCR and compared their SSCP profiles with standard strains.</p><p><b>RESULTS</b>67 strains of streptomycin, isoniazid or rifampicin resistant Mycobacteria tuberculosis were found in routine drug resistance test, with the percentages of 80.5% (54/67), 58.2% (39/67) respectively. PCR-SSCP showed that out of 67 drug-resistant strains, 66(98.5%) of rpsL, 47(70.1%) of rpoB and 42(62.7%) of KatG appeared abnormal.</p><p><b>CONCLUSION</b>Most of the resistant strains appeared gene mutation. The mution rates were higher than the results from routine drug resistance test.</p>
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Humanos , Carbón Mineral , Farmacorresistencia Bacteriana , Mutación , Mycobacterium tuberculosis , Genética , Reacción en Cadena de la Polimerasa , Polimorfismo Conformacional Retorcido-Simple , Silicotuberculosis , Microbiología , Esputo , MicrobiologíaRESUMEN
Long Jing 1 (L01) is the effective component extracted from asiatic todd-alia. In the experiments on rat thoracic aortic rings, L01 45-405 ?mol/L inhibited thecontraction initiated by high K~+ or NE, the IC_(50) value was 206.93 and 94.18 ?mol/Lrespectively. L01 shifted the dose-response curve of KCl or NE to the right, and reducedthe maximal response, also shifted that of CaCl_2 to the right parallelly. The effects ofL01 were similar to that of Ver (verapamil) in the blockage on PDC. Morever L01 80 ?mol/Linhibited the release of intracellular Ca~(2+) and extracellular Ca~(2+) influx initiated by NE,the former effect was more significant than the latter (this was different from Ver). Theseresults suggested that L01 was probably a new calcium antagonist different from Ver.