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As one of the "Three Drugs Three Prescriptions" anti-COVID-19 traditional Chinese medicine, Jinhua Qinggan granules (JHQG) has been proved to have clear clinical effects. With complex medicinal flavors and ingredients, there is no systematic research report on chemical composition in vivo or in vitro. An ultrahigh pressure liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-QTOF/MS) method was developed in this study to identify the components of the anti-COVID-19 traditional Chinese medicine JHQG granules. Analyze the collected rat plasma samples after administration and explore the exposed components in rats within 8 hours after intragastric administration. Preliminary pharmacokinetic analysis was then performed on this basis. Through UPLC-QTOF/MS analysis and verification by standard products, a total of 77 chemical components in JHQG formula have been identified, among which 22 compounds were highly exposed in vivo, mainly derived from three medicinal materials of honeysuckle, scutellaria and forsythia. Through the assessment of the blood drug concentration by the compartment model, 6 PK parameters of 4 high-exposure chemical components have been obtained, clarifying the metabolic characteristics of the main exposed components in JHQG briefly. The method is simple, efficient, sensitive and accurate and provides research basis to the clarification of the pharmacodynamics material basis and mechanism of JHQG, which has certain reference significance for the basics and applications research of the traditional Chinese medicine prescriptions in fighting the SARS-CoV-2.
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@#【Objective】The aim of this study is to investigate whether Fuzi polysaccharide（FPS）inhibits calcification of vascular smooth muscle cells（VSMC）and its underlying mechanism involving ceramide signaling.【Methods】We used Ox- LDL to induce in vitro model of human VSMC calcification in this study. FPS at different concentrations was used to treat human VSMC. Cell calcification was assessed by alizarin red staining. The mRNA expressions of osteogenic differentiation markers including Msx2，Osterix and BMP2，and contractile marker SMA were analyzed by qRT- PCR. The protein expressions of Msx2 and BMP2 were analyzed by western blot. Cell apoptosis was examined by TUNEL. Additionally，we investigated the effect of FPS on ceramide levels and N- SMase activity in VSMC. 【Results】We found that FPS inhibits Ox- LDL- induced VSMC apoptosis and calcification. Ceramide participates in Ox- LDL- induced apoptosis and calcification of VSMC. FPS reduces N- SMase activity and ceramide levels in Ox- LDL- treated VSMC. Collectively ， reducing N-SMase activity and ceramide levels could become a promising strategy for the treatment of vascular calcification.【Conclusion】We demonstrate that FPS attenuates VSMC calcification via targeting ceramide signaling.
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[Objective] Vascular calcification is a gene-regulated biological process similar to bone formation.It is very common in the patients with chronic kidney disease.Neutral sphingomyelinase 2 (nSMase2) is a key regulator of bone development,and is responsible for ceramide generation through sphingomyelin hydrolysis,but the role of nSMase2 in vascular calcification remains unclear.The aim of this study is to determine whether nSMase2 regulates high calcium and phosphate-induced calcification of vascular smooth muscle cells (VSMC).[Methods] In vitro model of human VSMC calcification was used in this study and high calcium and phosphate were used to induce calcification of VSMCs.GW4869 was used to inhibit nSMase2 activity and nSMase2 was knockdowned in cultured VSMC using nSMase2 siRNA.The expression of Runx2,BMP2 and Osterix was analyzed by qRT-PCR and calcification was assessed by alizarin red staining.[Results] We found that nSMase2 expression and ceramide levels were increased in the process of VSMC calcification (P＜0.05).Inhibition of nSMase2 activity by GW4869 and knockdown of nSMase2 attenuated high calcium and phosphate-induced VSMC calcification and down-regulated the expression level of Runx2,BMP2 and Osterix (P＜0.05).By contrast,ceramide accelerated rat VSMC calcification and increased ALP activity (P＜0.05).[Conclusion] We demonstrate that nSMase2/ceramide promotes high calcium and phosphate-induced VSMC calcification,suggesting that nSMase2/ceramide could participate in the progression of vascular calcification in patients with chronic kidney disease.
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Background Endophthalmitis is a serious,sight-threatening condition.Identifying the causative microorganisms is very important for available treatment of endophthalmitis.Objective This survey was to analyze the spectrum of organisms causing culture-proven endophthalmitis and their sensitivities to commonly antimicrobial agents.Methods Medical data of patients with culture-proven endophthalmitis at Zhongshan Ophthalmic Center from January 2003 through December 2010 were respectively reviewed.The outcomes included intravitreal isolates and antibiotic sensitivities were analyzed.Results Four hundred and sixty-nine strains of organisms were isolated from 447 eyes of 447 patients with infective endophthalmitis,including 22 eyes of polymicrobial infection.In the organisms,gram-positive organisms were 241 (51.4％),fungi were 125 (26.7％) and gram-negative organisms were 103 (22.0％).The most common organisms were Staphylococcus epidermidis in 29.4％,Aspergillus in 7.7％ and Pseudomonas aeruginosa in 5.3％.In this group of infective patients,the most common clinic settings were posttraumatic endophthalmitis (72.7％),and then were postoperative endophthalmitis (10.5％),endogenous endophthalmitis (9.8％) and keratitis (6.9％).Most gram-positive organism and gram-negative organism were sensitive to levofloxacin and cefoperazone.The susceptibility rate of gram-positive organism to chloromycetin was increased in 2007-2010 years compared with 2003-2006 years (x2=5.398,P＜0.05).The susceptibility rate to ciprofloxacin of gram-negative organisms declined (x2 =5.398,P ＜ 0.05),but that to rifampicin increased in the duration of 2007-2010 compared with 2003-2006 (x2 =4.500,P ＜ 0.05).Conclusions Gram-positive organisms are the most commonly causative organisms of endophthalmitis.Most bacterial organisms are sensitive to levofloxacin and cefoperazone.Local data of culturing and susceptibility test offers a guideline for the treatment of infectious endophthalmitis.
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<p><b>OBJECTIVE</b>To study the therapeutic efficacy of siRNA fragments silencing p75 neurotrophin receptor (p75(NTR)), which may be a key regulator of glioma cell apoptosis and invasion.</p><p><b>METHODS</b>The siRNA sequence fragments targeting p75(NTR) were designed and transferred into human glioma cell line U251. RT-PCR and immunocytochemistry method were used to explore the expression of p75(NTR) mRNA and protein. Cell adhesion assay was employed to detect cellular adhesion ability, and soft agar clone formation assay was adopted to identify oncogenicity, and a U251 glioma model was established in nude mice. The intracranial tumor volume was detected by MRI. The expression of p75(NTR), NGF and cyclin D2 were identified using immunohistochemistry. Cell apoptosis was detected by apoptosis kit in situ.</p><p><b>RESULTS</b>The siRNA fragments targeting p75(NTR) were capable of decreasing mRNA and protein expression of p75(NTR) in U251 glioma cell line. Both the cellular adhesion ability and oncogenicity were weakly relevant. The p75(NTR) expression level was negatively correlated with cyclin D2 and apoptosis, and positively correlated with NGF expression. The siRNA sequence fragments targeting p75(NTR) were effective in decreasing the gross volume of tumor; prolonged the survival time of mice, and the edge of tumor was much sharper than that of the control group.</p><p><b>CONCLUSIONS</b>The gene silencing technique by siRNA targeting p75(NTR) is capable of decreasing tumor invasion and cell proliferation as well as inducing cell apoptosis. It is expected to be a new choice for glioma gene therapy.</p>