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As an effective prescription for the treatment of rheumatoid arthritis (RA), Huangqin Qingre Chubi capsule (HQC) is still blank in quality control. This study aims to explore quality markers (Q-markers) for HQC in the treatment of RA by integrating network pharmacology and pharmacokinetics. By constructing the visualization network of "pharmacodynamic ingredient-target-pathway", the potential Q-Marker of HQC treatment for RA was preliminatively predicted. A rat model of rheumatic heat obstruction syndrome collagene-induced arthritis (CIA) was established to elucidate the dynamic quantification law of pharmacodynamic components of HQC in the disease state of rats. To establish the inflammatory model of RA synovial fibroblasts (MH7A) induced by tumor necrosis factor-α (TNF-α) in vitro. The effects of active ingredients on protein expression of sphingosin kinase-1 (Sphk1) and p-SphK1 were detected. The network pharmacological results showed that baicalin, geniposide, luteolin, coixol and amygdalin were the important active components of HQC treatment for RA. Quantitative analysis results further verified the measurability of these five components. The expression of Sphk1 and p-SphK1 was significantly inhibited by geniposide and baicalin by Western blotting. The above studies determined that the above 5 components could be used as Q-markers in the treatment of RA by HQC. This experiment was approved by the Experimental Animal Ethics Committee of Anhui University of Chinese Medicine (approval number: AHUCM-rats-2021049). All procedures were conducted in strict accordance with the principles of animal use and care.
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Objective: To establish a dynamic syndromic surveillance system in the border areas of Yunnan Province based on information technology, evaluate its effectiveness and timeliness in the response to common communicable disease epidemics and improve the communicable disease prevention and control in border areas. Methods: Three border counties were selected for full coverage as study areas, and dynamic surveillance for 14 symptoms and 6 syndromes were conducted in medical institutions, the daily collection of information about students' school absence in primary schools and febrile illness in inbound people at border ports were conducted in these counties from January 2016 to February 2018 to establish an early warning system based on mobile phone and computer platform for a field experimental study. Results: With syndromes of rash, influenza-like illness and the numbers of primary school absence, the most common communicable disease events, such as hand foot and mouth disease, influenza and chickenpox, can be identified 1-5 days in advance by using EARS-3C and Kulldorff time-space scanning models with high sensitivity and specificity. The system is easy to use with strong security and feasibility. All the information and the warning alerts are released in the form of interactive charts and visual maps, which can facilitate the timely response. Conclusions: This system is highly effective and easy to operate in the detection of possible outbreaks of common communicable diseases in border areas in real time, so the timely and effective intervention can be conducted to reduce the risk of local and cross-border communicable disease outbreaks. It has practical application value.
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Humanos , Influenza Humana , Vigilância de Evento Sentinela , Síndrome , China , Telefone CelularRESUMO
Rheumatoid arthritis (RA) is an autoimmune disease with angiogenesis, inflammatory factor infiltration and joint destruction as the main pathological features. Angiogenesis promotes the development of RA and plays an important role in its pathogenesis. The hypoxia-inducible factor (HIF)-vascular endothelial growth factor (VEGF)-angiopoietin-2 (Ang-2) signal transduction is a critical pathway to induce synovial angiogenesis. Targeting HIF-VEGF-Ang-2 signal transduction to inhibit synovial angiogenesis is a promising approach for RA treatment. This article reviews the role and mechanism of HIF-VEGF-Ang-2 signal transduction-mediated synovial angiogenesis in RA, in order to provide a new target and strategy for RA treatment.
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Objective To investigate whether NO can relieve hypertensive cerebrovascular and renal injury by regulating the ratio of peripheral blood T lymphocyte subsets and reducing proinflammatory cytokine release . Methods Twelve-week SHR and WKY rats were randomly divided into three groups :WKY group ,SHR group and SHR+NO intervention group ,with 6 rats in each group .Rats in SHR+ NO intervention group were injected intraperitoneally with sodium nitroprusside according to 10 μg/(kg · d) for 4 weeks ,while WKY group and SHR group were injected intraperitoneally with an equal amount of saline for 4 weeks .After measuring the tail artery blood pressure ,basilar artery and renal pathological changes were observed by HE staining ;the rates of CD4+ /CD8+ and CD4+ CD25+ T cells were detected by flow cytometry ;and the expression of TNF-α was detected by ELISA .Results The expressions of CD4+ /CD8+ and TNF-αin SHR group were significantly higher than those in WKY group (P<0 .01) while the rate of CD4+ CD25+ in SHR group was significantly lower than that in WKY group (P<0 .01) .The expressions of CD4+ /CD8+ and TNF-α in SHR+ NO intervention group were significantly lower than those in SHR group whereas the rate of CD4+ CD25+ was significantly higher than that in SHR group (P<0 .05) .Conclusion NO can improve the target organ damage caused by hypertension by regulating the peripheral blood T lymphocyte subsets ratio and reducing the release of proinflammatory factors .
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Objective To investigate the expression of transmembrane protein 16A (TMEM16A)in the cochlea of guinea pigs and its relationship with the age-related hearing loss.Methods We used auditory brainstem response (ABR)to explore the changes of hearing in guinea pigs of different age (groups of 2 w,3 m,1 y,and D-galactose).The distribution and expression of TMEM16A in the cochlea were detected by immunofluorescence and Western blot.Results ABR threshold was gradually increased,with significant difference between D-gal and the other three groups (P <0.01).TMEM16A was expressed in the cochlear striae vascularis at different ages,and the expression increased with age before 1 y (P <0.05). However, its level was increased in D-gal group and significantly differed from that in 3 m and 1 y groups (P < 0.05 ).Conclusion The change in TMEM16A expression in the cochlear striae vascularis of guinea pigs may be related to age-related hearing loss.
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Objective To investigate the expression of transmembrane protein 16A (TMEM16A)in the cochlea of guinea pigs and its relationship with the age-related hearing loss.Methods We used auditory brainstem response (ABR)to explore the changes of hearing in guinea pigs of different age (groups of 2 w,3 m,1 y,and D-galactose).The distribution and expression of TMEM16A in the cochlea were detected by immunofluorescence and Western blot.Results ABR threshold was gradually increased,with significant difference between D-gal and the other three groups (P <0.01).TMEM16A was expressed in the cochlear striae vascularis at different ages,and the expression increased with age before 1 y (P <0.05). However, its level was increased in D-gal group and significantly differed from that in 3 m and 1 y groups (P < 0.05 ).Conclusion The change in TMEM16A expression in the cochlear striae vascularis of guinea pigs may be related to age-related hearing loss.
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Children are a special group in clinical medical treatment. The significant individual differences require individualized administration of drugs. In recent years, the rapid development of population pharmacokinetics which are widely used in pharmaceutical research is an effective method to achieve individual administration. At home and abroad a large number of population pharmacokinetics are researched in chidren for drugs with narrow therapeutic window or in children with significant individual differences, which can provide information for the individualized dosing of drugs.
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Children are a special group in clinical medical treatment. The significant individual differences require individual?ized administration of drugs. In recent years,the rapid development of population pharmacokinetics which are widely used in pharma?ceutical research is an effective method to achieve individual administration. At home and abroad a large number of population pharma?cokinetics are researched in chidren for drugs with narrow therapeutic window or in children with significant individual differences , which can provide information for the individualized dosing of drugs.
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The Wnt pathway plays a crucial role in skeletal development and is indispensable for determination of OS cell lines. In recent years, experimental evidences on Wnt signaling pathway in OS cell lines and OS animal models, and that of Wnt signaling pathway as a diagnosis and prognosis marker of OS suggested that Wnt signaling pathway plays an important role in the progression, invasion and metastasis of OS. In addition, the strategy and safety evaluation to target Wnt to treat OS are underway. Exploring the significant role of Wnt signaling pathway in OS may aid in personalizing therapeutics to increase patient survival.
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Chymosin is an important industrial enzyme widely used in cheese manufacture. To improve expression efficiency of recombinant bovine chymosin in Kluyveromyces lactis strain GG799, we designed and synthesized a DNA sequence encoding bovine prochymosin gene (GenBank Accession No. AA30448) by using optimized codons. The synthesized prochymosin gene was amplified by two-step PCR method, and then cloned into the expression vector pKLAC1, resulting in pKLAC1-Prochy. pKLAC1-Prochy was linearized and transformed into K. lactis GG799 by electrotransformation. Positive clones were screened by YEPD plates containing 1% casein. A recombinant strain chyl with highest activities and multi-copy integration which was detected by using specifical integration primers was chosen and fermented in flask. Prochymosin was expressed in K. lactis successfully. SDS-PAGE analysis revealed that the purified recombinant bovine prochymosin had a molecular mass of 41 kDa. After acid treatment, molecular weight of chymosin is about 36 kDa, the same as native bovine chymosin. Activity tests showed that the chymosin activity of the culture supernatant was 99.67 SU/mL after 96 h cultivation. The activities of chymosin were not prominent increased when galactose was used as carbon source instead of glucose, which proved that the fermentation of recombinant strain does not need galactose inducing. The recombinant K. lactis strain obtained in this study could be further used to produce recombinant chymosin for cheese making.
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Animais , Bovinos , Quimosina , Genética , Precursores Enzimáticos , Genética , Regulação Fúngica da Expressão Gênica , Genética , Vetores Genéticos , Genética , Kluyveromyces , Genética , Metabolismo , Engenharia de Proteínas , Proteínas Recombinantes , GenéticaRESUMO
In this study, we efficiently expressed the active antimicrobial peptide (CAD), which fused with the site-mutated coat protein (EDDIE) of the classical swine fever virus, in Escherichia coli. First, we obtained the e-cad fusion gene from the CAD gene and the EDDIE gene using overlapping PCR. Then to get the recombinant expression vector (pETED), the e-cad fusion gene was cloned into the pET30a vector by a site-directed homologous recombination technique. The EDDIE-CAD fusion protein expressed in E. coli as inclusion bodies, and its yield was more than 40% of total bacterial proteins. After renaturated in vitro and self-cleavage of the fusion protein, we obtained the antimicrobial peptide Cecropin AD. Antimicrobial experiments showed that the Cecropin AD efficiently inhibited the growth of G+ and G- bacteria, but it weakly inhibited the growth of Saccharomyces. This method provides an excellent way for high expression of antimicrobial peptides when fused with EDDIE.