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Objective@#To observe the effect of tumor necrosis factor-α (TNF-α) monoclonal antibody on autophagy in allergic rhinitis (AR) mice.@*Methods@#Thirty six weeks old BALB/c mice were randomly divided by random number table method into five groups: control group, model group (AR group), TNF-α antibody intervention group (AR+TNF-α group), autophagy inhibitor (3-methylindole, 3-NA) intervention group (AR+3-MA group), TNF-α antibody combined with autophagy inducer rapamycin (RAP) intervention group (AR+TNF-α+RAP group), with 6 mice in each group. AR model was established by conventional method, the corresponding reagent was administered before nasal cavity stimulation sensitization and during the whole experiment. Behavioral scores of mice were obtained, blood was collected from the eye socket, and mice in each group were sacrificed to collect nasal mucosa tissue samples. Pathological changes of nasal mucosa were observed by hematoxylin-eosin staining. Expression levels of inflammatory factor and IgE in serum were detected by enzyme-linked immunosorbent assay (ELISA). Expressions of autophagy related indicators microtubule-associated protein-1 light chain-3B (LC3B), Beclin-1, sequestosome1 (p62), autophagy-related 5 (ATG5), autophagy-related 7 (ATG7) were measured by Real-time PCR and Western blot. The aggregation of LC3B protein was observed by immunofluorescence. SPSS 19.0 software was used for statistical analysis.@*Results@#Compared with the AR model group, symptoms of AR in AR+TNF-α group and AR+3-MA group were mild; the pathological changes of nasal mucosa were weak; the expression of IgE, TNF-α, interleukin 4 (IL-4), interferon-γ (IFN-γ) in serum significantly reduced (IgE: 666.19±78.35 (±s) vs. 692.38±64.29 vs. 1 059.05±146.44, TNF-α: 112.06±12.95 vs. 113.17±15.43 vs. 161.22±17.96, IL-4: 54.05±7.14 vs. 58.26±5.67 vs. 79.95±6.33, IFN-γ: 28.58±4.51 vs. 30.67±2.60 vs. 39.83±3.31, all P<0.05), and the expression of LC3B Ⅱ/Ⅰ, Beclin-1, ATG5, ATG7 in nasal mucosa significantly decreased, the expression of p62 significantly elevated. After intervention with autophagy inducer RAP, the therapeutic effect of TNF-α monoclonal antibodies on AR was antagonized.@*Conclusion@#TNF-α monoclonal antibody significantly improves nasal symptoms in AR mice by inhibiting autophagy levels.
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MicroRNA (miRNA),sharing the character of regulating the transcriptional level and expression level of mRNAs,is one kind of small non-coded RNAs.At present,innate immune has become one of the hot topics for researchers,and miRNAs as a sort of bioactive substance greatly take part in the whole regulation progress.In detailed,miRNAs can influence the immune state of immune cells during innate immune period and further regulate inflammatory conditions in whole body.By systematically summarizing miRNA function during innate immunity,this present review may provide a reference for peer researchers.
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This study aimed to detect the combined effects of gallic acid (GA)and ciprofloxacin (CIP)on the murine chronic rhinosinusitis(CRS)model in mice.Pseudomonas aeruginosa from refractory CRS nasal samples were isolated and a CRS model in mice was induced.GA and CIP were intragastrically administered singly or in combination.The nasal histopathologic change was observed by hematoxylin and eosin (HE)staining.The concentration of TNF-α;IL-6 and IL-8 in serum were determined by ELISA assay.The activity of SOD and contents of MDA and ROS were measured with commercially available kits.The expressions of IκB;NF-κB p65;TNF-α;IL-6 and IL-8 in nasal mucosa tissues were measured by Western blotting assay.The results showed that the inflammation of CRS in each treatment group was significantly attenuated.The expression level of TNF-α;IL-6;IL-8;MDA;NF-κB p65 and the contents of ROS were reduced significantly in treated groups;while the activity of SOD and the expression level of IκB were increased.More obvious effects were achieved in CA and CIP combined group.The data showed that combination of GA and CIP was superior to GA or CIP alone;and the combined therapy might be related with inhibiting NF-κB signaling pathway and downregulating the expressions of TNF-α;IL-6 and IL-8.
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Objective To characterize Hfq-dependent phenotypes in stress response and to dissect Hfq-dependent transcription of virulence genes and stress-responsive genes in Vibrio cholera.Methods The hfq null mutant strain (△hfq) and the complemented mutant strain (△hfq/pUC18-hfq) were constructed from the wild-type Vibrio cholera.Comparisons on the motility,biofilm formation,growth under various oxygen-supplying conditions,outer membrane resistance,and sensitivity to oxidative stress were analyzed between the wild type strain and the mutant strains.Reverse-transcript fluorescence quantitative PCR (RT-qPCR) was used to determine the transcriptional levels of target genes in the above mentioned strains.Results △hfq and △hfq/pUC18-hfq strains were successfully constructed.The motility,outer membrane resistance and sensitivity to oxidative stress were reduced,but biofilm formation was enhanced in the hfq null mutant strain.RT-qPCR testified that Hfq had regulation effects on gene transcription for forming falagellum,extracellular polysaccharide,outer membrane protein and oxidative stress in Vibrio cholera.Conclusion As a RNA chaperone,Hfq could affect Vibrio cholera in its biofilm formation,resistance to oxidative stress and antibiotics resistance through regulating the transcription of multiple metabolic genes and virulence genes,which indicates that Hfq,combined with other regulators,may play a key role in the complex regulation of metabolic genes and virulence genes.
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As a traditional Chinese medicine, Valeriana jatamansi has a long history of application in China. It is widely distributed and commonly adopted by many ethnic groups. In particular, its roots have a wide range of medicinal value. With the increasingly more attention on it from domestic and foreign researchers, there have been more and more studies on its pharmacological activity and mechanism. This essay summarizes domestic and foreign reports on its pharmacological activity and mechanism.
Subject(s)
Animals , Humans , Anti-Infective Agents , Pharmacology , Antihypertensive Agents , Pharmacology , Antineoplastic Agents, Phytogenic , Pharmacology , Central Nervous System Depressants , Pharmacology , Gastrointestinal Tract , Plant Extracts , Pharmacology , ValerianABSTRACT
Bacterial cellulose (BC) was prepared by Acetobacter xylinum in static culture. After purified by chemical treatment, the microstructure, chemical structure, crystal structure and mechanical property of BC were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffractometry (XRD) and tensile strength measurement respectively, and compared with those of the imported bacterial cellulose wound dressing served as control sample (XBC). The results indicated that the diameter of the BC was (22 +/- 9) nm, and the crystallinity index was 89.71%. The tensile strength and the Young's mouduls of BC were significant higher than XBC both in wet and dry states. The biocompatibility of BC and XBC were evaluated by cytotoxicity test, delayed contact sensitization study in the Guinea Pig and skin irritation test. The results showed that BC had reliable biocompatibility as well as XBC. With the unique nanostructure, high crystallinity, high mechanical strength, and reliable biocompatibility, BC produced in our country as well as XBC can be used as a safe biomaterial for the medical applications.
Subject(s)
Animals , Biocompatible Materials , Chemistry , Cellulose , Chemistry , Culture Techniques , Gluconacetobacter xylinus , Metabolism , Guinea Pigs , Materials Testing , Nanoparticles , Tensile StrengthABSTRACT
OBJECTIVE To study the genotype distribution of CTX-M type ESBLs in ESBLs-producing Escherichia coli. METHODS All 138 strains of E. coli were collected from the First and Third Affiliated Hospitals,Zhengzhou University from 2006 to 2007. The CTX-M type ESBLs genes were amplified by PCR,the products were sequenced after purification. RESULTS Eighty-two of 138 ESBLs-producing E. coli isolates were classified with CTX-M type-ESBLs genes,and there were 53,27 and 2 strains contained only one type,two types and three types of CTX-M ESBLs,respectively. Among CTX-M type-ESBLs genes,they were identified with CTX-M-1(43 strains),CTX-M-14(56 strains),CTX-M-25(7 strains) and CTX-M-38 (5 strains). CONCLUSIONS The most popular type of CTX-M ESBLs in E. coli is CTX-M14 in Zhengzhou,and two or three kinds of genotypes can exist in 1 strain.
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Human cytomegalovirus (HCMV) is the most common cause of congenital infection, resulting in birth defects such as microcephaly. In this study, RT-PCR and Western Blotting were performed to quantify the regulation of endogenic nerve growth factor expression in neuroglia cells by HCMV infection. The results showed that basal, endogenous NGF expression in U251 was unchanged during early HCMV infection. NGF expression is strongly down-regulated during the latent phase of infection. These results suggest that HCMV can depress the NGF expression in U251 cells.
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Objective: To express CTX-M-38 type extended-spectrum-lactamase, and detect its distribution and antibiotic susceptibilities. Methods: Total of 46 strains producing ESBL E.coli was collected from the first affiliated hospital of Zheng zhou University. The CTX-M-38 ESBL gene was selected by PCR using gene recombination technique to construct pET28a-CTX-M-38. The expression of CTX-M-38 in BL21 E.coli and its antibiotic susceptibilities were carried out by liquid dilution test. Testing the enzyme activities of culture supernatant and bacteria sonicate to reflect its distribution. Results: The size of amplified gene product was about 900 bp. The DNA sequence was matched with the information of gene bank. The enzyme activities from bacteria sonicate were stronger than the culture supernatant .The transformant was resistance to penicillins, the first, second and third generations of cephalosporins. It was sensitive to imipenem. The transformant was also sensitive to ceftazidime and aztreonam in vitro, and resistance to antibiotics including beta-lactamase inhibitors except piperacillin/tazobactam. The transformant was also resistance to gentamicin,minocycline, ciprofloxacin and levofloxacin. Conclusion: The CTX-M-38 type ESBL is successfully expressed at designed experimental condition in this study. The product mainly lies inside the bacteria. The transformant shows wide resistance to antibiotics.
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OBJECTIVE To develop a macroarray method to detect pathogens in cerebrospinal fluid.METHODS According to the bacterial 16S rRNA genes,designed 10 kinds of specific probes and a pair of universal primers that can amplify rRNA gene of all bacteria.The tailed probes were spotted onto a nylon membrane.DNA was isolated from each pathogen,and subjected to UP-PCR to amplify target fragments,which were labeled with bio-16-dUTP at the same time.All those denatured fragments were hybridized to the probes on nylon membrane and visualized by AKP labeled avidin.The sensitivity and specificity of the system were detected.A total of 32 CSF samples,which were verified the bacterial infection by the routine method,were tested by this method.RESULTS It was sensitive to 10 CFU/ml when detecting Escherichia coli.Every kind of pathogens only reacted to its corresponding probes fixed on nylon membranes,which showed high specificity.The result of identifying 32 CSF clinical specimens accorded with that of routine method.CONCLUSIONS The method can screen out common pathogens in CSF sensitively and exactly.
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Chinese materia medica (CMM) has double complexity in bioactive ingredient and its mechanism. It is difficult to explain by the modern biomedicine theory So it seriously restricts the modernization of CMM The modern CMM should have the high quality standard to meet the needs of international standard It can be guaranteed by spreading the GAP for Chinese medicinal materials and GMP for standard production The mechanism depends on using the DNA microarray to set up “the gene expression difference chart”, to study on the combination of CMM and gene expression difference chart Meanwhile, we can establish a totally new method of screening modern CMM based on the gene expression difference chart, it can really make the modernization and internationalization of CMM