ABSTRACT
In order to investigate the enzyme production mechanism of yak rumen-derived anaerobic fungus Orpinomyces sp. YF3 under the induction of different carbon sources, anaerobic culture tubes were used for in vitro fermentation. 8 g/L of glucose (Glu), filter paper (Flp) and avicel (Avi) were respectively added to 10 mL of basic culture medium as the sole carbon source. The activity of fiber-degrading enzyme and the concentration of volatile fatty acid in the fermentation liquid were detected, and the enzyme producing mechanism of Orpinomyces sp. YF3 was explored by transcriptomics. It was found that, in glucose-induced fermentation solution, the activities of carboxymethyl cellulase, microcrystalline cellulase, filter paper enzyme, xylanase and the proportion of acetate were significantly increased (P < 0.05), the proportion of propionate, butyrate, isobutyrate were significantly decreased (P < 0.05). The results of transcriptome analysis showed that there were 5 949 differentially expressed genes (DEGs) between the Glu group and the Flp group, 10 970 DEGs between the Glu group and the Avi group, and 6 057 DEGs between the Flp group and the Avi group. It was found that the DEGs associated with fiber degrading enzymes were significantly up-regulated in the Glu group. Gene ontology (GO) function enrichment analysis identified that DEGs were mainly associated with the xylan catabolic process, hemicellulose metabolic process, β-glucan metabolic process, cellulase activity, endo-1,4-β-xylanase activity, cell wall polysaccharide metabolic process, carbohydrate catabolic process, glucan catabolic process and carbohydrate metabolic process. Moreover, the differentially expressed pathways associated with fiber degrading enzymes enriched by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were mainly starch and sucrose metabolic pathways and other glycan degradation pathways. In conclusion, Orpinomyces sp. YF3 with glucose as carbon source substrate significantly increased the activity of cellulose degrading enzyme and the proportion of acetate, decreased the proportion of propionate, butyrate and isobutyrate. Furthermore, the degradation ability and energy utilization efficiency of fungus in the presence of glucose were improved by means of regulating the expression of cellulose degrading enzyme gene and participating in starch and sucrose metabolism pathway, and other glycan degradation pathways, which provides a theoretical basis for the application of Orpinomyces sp. YF3 in practical production and facilitates the application of Orpinomyces sp. YF3 in the future.
Subject(s)
Animals , Cattle , Neocallimastigales/metabolism , Anaerobiosis , Rumen/microbiology , Propionates/metabolism , Isobutyrates/metabolism , Cellulose/metabolism , Fungi , Starch/metabolism , Glucose/metabolism , Acetates , Sucrose/metabolism , Cellulases , CellulaseABSTRACT
Objective To establish HPLC fingerprint of Xinjiaxiangruyin standard decoction soasto provide an analytical method for the quality control of Xinjiaxiangruyin granules. Methods HPLC was conducted on a phenonmenex Luna C18 column us?ing acetonitrile-0.1%H3PO4 in gradient elution modes as mobile phase,where detection wavelength was 280 nm and detection time was 130 min. With chlorogenic acid as reference peak,the Edition 2012 ofchromatographic fingerprint similarity evaluation systemsoftware was used to establish the standard decoction reference fingerprint,which was then compared with the fingerprint of 5 batchs of the granules for similarity evaluation. Results The similarity of the fingerprints between the 5 batches of granules and the standard decoction was higher than 0.90. Conclusion The method is simple,stable and reproducible,which could be used for the quality control of the granules as an adjuvant method.
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Objective:To investigate the expression of miR-16,miR-17,miR-30a etc in peripheral blood plasma ,mononuclear cells ( PBMC) and synovial fluid of patients with rheumatoid arthritis ( RA) and its clinical significance ,to provide a theoretical basis for the further research in the mechanism of RA.Methods:Collected 80 cases of RA patients ,32 cases of osteoarthritis ( OA) patients and 32 healthy human peripheral blood ,synovial fluid ,separating the plasma and PBMC;extraction of small RNA ,with specific stem loop primed reverse transcription into cDNA , establishing a mature miRNA-T carrier and making standard curve;stem-loop method Real-time quantitative PCR was adopted to detect the expression of miRNAs in plasma ,PBMC and synovial fluid,and for correlation analysis;and with RA activity monitoring indicators rheumatoid factor (RF),erythrocyte sedimentation rate (ESR),C-reactive protein (CRP) correlation analysis.Results:In RA group,the expression of miR-16,miR-17,miR-30a,miR-106,miR-101 and miR-130 in plasma,PBMC and synovial fluid were upregμlation compared with OA group,the healthy control group (P<0.05),but the expression level of miR-101 in plasma of RA and OA -group difference was not statistically significant.Correlation analysis showed that 6 kinds of miRNA in the plasma ,PBMC and joint fluid have varying degrees of positive correlation ( P<0.05 ).Correlation analysis showed that miRNA in plasma , PBMC and synovial fluid have one or more indicators of a positive correlation with RF , ESR, CRP ( P<0.05 ).Conclusion:The expression of miR-16,miR-17,miR-30a,miR-101,miR-106 and miR-130 is changes significant in the peripheral blood and synovial fluid of RA patients ,indicate the miRNAs may be through some related genes play an important role in the process of the pathogenesis of RA;its expression level can be used as an effective indicator of RA disease activity , and provide new diagnostic markers for RA.
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Objective:In order to investigate the role of microRNA in the pathogenesis of ankylosing spondylitis patients,we detected the peripheral blood of patients with ankylosing spondylitis in miR-17,miR-181,miR-106,miR-30 and miR-495 expression, thereby to study the role of microRNA regulation of clinical and diagnostic value in the pathogenesis of ankylosing spondylitis provide new ideas.Methods:Collection of patients with ankylosing spondylitis and normal peripheral blood,peripheral blood mononuclear cells were isolated and extracted PBMC small RNA,using primers specific stem-loop reverse transcribed into cDNA,and build a mature miR-17,miR-181,miR-106,miR-30 and miR-495 T-carrier,standard curve,the use of stem-loop method by Real-time PCR technology to detect miR-17,miR-181,miR-106,miR-30 and miR-495 expression level.Results: In this study,92 cases of clinical samples were collected,of which 61 patients with AS,normal 31 cases.By Real-time PCR detection showed that compared with normal subjects,there was upregulation of miR-106 (P>0.05) and miR-30 (P>0.05) in the peripheral blood of patients with ankylosing spondylitis;down-regulation were miR-181 ( P0.05 ) , in which the miR-181 and miR-495 was statistically significant.Conclusion:Compared with normal, there are differences in the peripheral blood of patients with ankylosing spondylitis expression of miR-495 and miR-181,which may be targeted to regulate TLR-4,HLA-B,DVL,GSK/3βgenes,this may be found in the pathogenesis of ankylosing spondylitis studies provide new ideas,to become a new clinical diagnosis markers.
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To detect the influence of rapamycin on the expression of miR-30b,miR-200a and miR-17-5p etc in macrophages and provide the basis to study the regulation of miRNA in autophagy mechanism of macrophages .Methods: Small RNA was extracted at different times after stimulated with rapamycin in cultured RAW 264.7 cells.After using the stem-loop reverse transcription primers to reverse transcribed into cDNA ,the expression of miR-30b ,miR-30c,miR-106a,miR-214,miR-183,miR-200a, miR-376c,miR-17-5p, miR-142-3p, miR-377 was detected by Real-Time PCR.Results: After RAW264.7 cells was treated by rapamycin for 2,4,6 and 8 hours,the expression of miR-17-5p and miR-106 increased (More than 2.1 times,P<0.05) in 2,4 and 6 hours;miR-214 was up regulated in 2 and 8 hours (More than 2.4 times,P<0.05);miR-30b,miR-30c,miR-183,miR-200a,miR-376c and miR-142-3p was up regulated in 2,6 and 8 hours (2.4 times,P<0.05 );while miR-183 and miR-200a was down regulated at 4 hours(More than 2.1 times,P<0.05);miR-30b was significantly low expression in 8 hours (more than 50 times,P<0.05);miR-377 was up regulated at 4 hours (more than 2.5 times,P<0.05),but was significantly down regulated at 2 and 8 hours (More than 50 times,P<0.05) Conclusion: The expression of miR-200a,miR-30b,miR-377,miR-30c,miR-376c and miR-17-5p is significantly changed after rapamycin stimulated RAW264.7 macrophages,indicated the miRNA may plays an important role in autophagy through the regulation of autophagy-related genes.
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Objective To investigate the effect of JUC on the incisions for episiotomy.Methods Three hundred primiparas to undergo episiotomy in our hospital were divided into two groups in equal number.The experiment group was given JUC Spray before suturing and the control group did not use any solution.In the two groups,antibiotics were not used after the operation,and the incisions were only cleaned with 0.5%povidone-iodine 2 times a day.Result There were significant differences between the two groups in terms of postoperative pains,inflammation and healing in the wounds,and hospital stay(P<0.05).Conclusions Application of JUC after episiotomy could be long-acting in antibacteria.It can reduce wound pain,improve wound healing rate, decrease the medical cost and shorten the hospital stay.
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Objective The regulation of neuroendocrine hormones on the innate immune responses remains controversial.This report investigated whether the adrenal-derived hormones directly enhanced the innate immune responses and alternation of main pattern recognition receptors in adrenalectomized rats following blast injury.Methods Bilateral adrenalectomy was performed on Sprague-Dawley adult rats,who inflicted with mild blast wave injury in 7 d later.Peripheral blood leukocyte counting,macrophage phagocytosis,bacterial translocation(BT) in lungs,livers and mesenteric lymph nodes(MLN) were measured and analyzed,and TNF-? level in the lung tissue was detected.The mRNA expressions of scavenger receptor-A(SR-A),CD14 and Toll like receptor-4(TLR-4) were assessed with RT-PCR.Results After blast injury,the number of peripheral leukocytes was decreased(P