Corilagin inhibits SARS-CoV-2 replication by targeting viral RNA-dependent RNA polymerase

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has become one major threat to human population health. The RNA-dependent RNA polymerase (RdRp) presents an ideal target of antivirals, whereas nucleoside analogs inhibitor is hindered by the proofreading activity of coronavirus. Herein, we report that corilagin (RAI-S-37) as a non-nucleoside inhibitor of SARS-CoV-2 RdRp, binds directly to RdRp, effectively inhibits the polymerase activity in both cell-free and cell-based assays, fully resists the proofreading activity and potently inhibits SARS-CoV-2 infection with a low 50% effective concentration (EC

Improvement Effects of Dantaojin Extract on Oxidative Stress Injury of Liver and Kidney in Chronic Lead Po i- soning Model Mice / 中国药房

OBJECTIVE：To stu dy the improvement effect s of Dantaojin （Salvia miltiorrhiza ，Persicae Semen ，Curcumae Radix）extract on oxidative stress of liver and kidney in chronic lead poisoning model mice. METHODS ：Totally 72 mice were randomly divided into normal control group ，model group ，positive control group （dimercaptosuccinate，70 mg/kg），Dantaojin extract low-dose ，medium-dose and high-dose groups （20，40，60 g/kg），with 12 mice in each group. Except for normal control group，other groups were given intraperitoneal injection of lead acetate solution 20 mg/kg（every other day ，consecutive 20 days） to establish chronic lead poisoning model. After modeling ，administration groups were given relevant medicine intragastrically ， normal control group and model group were given constant volume of water intragastrically ，once a day ，for consecutive 20 days. After last medication ，body weight of mice was weighed ，and organ coefficients （kidney and liver ）were calculated. Serum levels of ALT ，AST，BUN and Scr were determined by automatic biochemical detector. HE staining was used to observe histopathological features of liver and kidney. ELISA method was used to determine the levels of GSH-Px ，SDO and MDA in liver and renal tissue. RESULTS：Compared with normal control group ，body weight （except for high-dose group ），the levels of SOD （except for high-dose group ）and GSH-Px were all decreased significantly in model group and Dantaojin extract groups ，while the renal coefficients（except for high-dose group ），liver coefficients （except for low-dose ，medium-dose and high-dose group ），the levels of BUN （except for high-dose group ），Scr，AST（except for high-dose group ），ALT and MDA were increased significantly （P＜ 0.05 or P＜0.01）. The epithelial cells of glomerulus and renal tubules were atrophied ，the arrang ement of hepatocytes was loose and some cells were necrotic. Compared with model group ，body weight ，the levels of SOD and GSH-Px were increased significantly in positive control group and Dantaojin extract groups，while the renal and liver coefficients ，the levels of BUN，Scr，AST，ALT and M DA were decreased significantly（P＜0.05 or P＜0.01）. Histopathological fea tures of liver and renal tissue were improved significantly in Dantaojin extract medium-dose and high-dose groups. CONCLUSIONS ：Dantaojin extract could improve oxidant stress injury in liver and renal tissue，the mechanism of which may be associated with eliminating reactive oxygen radicals ，inhibiting lipid peroxidation and enhancing antioxidant defense ability.

Effects of donepezil on the expression of neurotrophic related factors in subventricular zone of cerebral infarction mice / 中华行为医学与脑科学杂志

Objective To investigate the effect of donepezil on subventricular zone ( SVZ) neuro-genesis related neurotrophic factors after cerebral infarction. Methods Mice were randomly assigned into three groups: vehicle-treated sham group (Sham+vehicle,n=18),vehicle-treated middle cerebral artery oc-clusion (MCAO) group (MCAO + vehicle,n=30) and donepezil-treated MCAO group (MCAO+donepezil, n=30). Middle cerebral artery occlusion( MCAO) was induced by thread-occlusion method. Nissl staining was used to measure the infarct volume and the modified neurological severity score(mNSS) was used to as-sess neurologic function and brain water content was detected to assess brain edema degree. Proliferative cells and neuroblasts were labeled with 5-bromodeoxyuridine ( BrdU) and doublecortin ( DCX). The SVZ BrdU+/DCX+cells were detected by immunofluorescence. The expression of glial cell line-derived neurotro-phic factor (GDNF),brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) were detec-ted by Western blot. Results The infarct volume of MCAO + donepezil group ((13. 33±4. 55)%) was sig-nificantly lower than that of MCAO + vehicle group ((31. 33±3. 93)%,t=7. 34,P<0. 05). The neurologic deficits were significantly ameliorated after donepezil treatment,and the brain water content of MCAO + done-pezil group ((71. 82±10. 18)%)was significantly less than that of MCAO + vehicle group ((85. 93± 7. 54)%,F=13. 480,P<0. 05). All differences were statistically significant (P<0. 05). The area of BrdU+/DCX+cells within SVZ of MCAO + vehicle group ((6. 16±1. 79)%) was significantly larger than that of sham + vehicle group ((2. 25±1. 09)%),and was fewer than that of MCAO+donepezil group ((16. 19± 2. 16)%,F=102. 756,P<0. 05). MCAO significantly promoted the expression of GDNF,BDNF and NGF within SVZ compared with sham operation,and donepezil increased these protein levels(F=15. 114,27. 121, 27. 398,P<0. 05). Conclusion Donepezil regulates neurogenesis via increasesing the expression of GDNF, BDNF and NGF within SVZ after cerebral infarction.

Research progress and mechanism of Galectin-9 and various malignant tumors / 实用肿瘤学杂志

Galectin-9 is a member of galectin,with chemotaxis eosinophils,regulating cell aggregation and adhesion,im-mune regulation and other functions.In recent years,many studies have shown that Galectin-9 plays an important role in a variety of malignant tumors,affecting tumor cell proliferation,apoptosis and other activities.This article focuses on the relationship between Ga-lectin-9 and various malignant tumors,and explores its possible mechanism of action.

Study on the Method of Microbial Limit Test for Preparation Liquid of Iron Sucrose Injection before Filtration and Sterilization / 中国药房

OBJECTIVE:To establish a method of microbial limit test for liquid preparation of Iron sucrose injection before filtration and sterilization.METHODS:According to the microbial limit test in the 1005 and general rules 1006 of 2015 edition of Chinese Pharmacopoeia (vol.Ⅴ),plate method and membrane filtration method were used to measure total number of aerobic bacteria (Staphylococcus aureus,Pseudomonas aeruginosa,Bacillus subtilis,Candida albicans,Aspergillus niger) and total number of molds and yeasts (C.albicans,A.niger).The optimal test method was obtained by comparing the bacterial recoveries.RESULTS:By plate method,the recoveries of P aeruginosa and B.subtilis were 2％ and 5％.The test sample was diluted 10 times with pH 7.0 sodium chloride-peptone buffer solution,and the bacterial recoveries were in the range of 88％ to 96％;but he medium was dark in color.By membrane filtration method,without rinse solution,the bacterial recoveries in the range of 88％ to 95％.Add rinse solution,the bacterial recoveries were in the range of 91％ to 103％.After validated,the recoveries of menbrane filtration method with tlushing fluid ranged 50％-200 ％,which was in line with the requnements.CONCLUSIONS:The membrane filtration method established in this experiment has higher bacterial recovery rate than the plate method.The bacterial recoveries rate were higher after adding rinse solution,and no dark substance in the surface of filter membrane affect the accotmt.It can be used as the microbial limit test method for preparation liquid of Iron sucrose injection before filtration and sterilization.

Identification and characterization of loop7 motif and its role in regulating biological function of human APOBEC3G through molecular modeling and biological assay

Human APOBEC3G (hA3G) is a cytidine deaminase which inhibits HIV-1 replication. The HIV-1 accessory protein viral infectivity factor (Vif) counteracts with hA3G by targeting it for proteasomal degradation. In this work, we constructed and optimized molecular models of the hA3G dimer and the hA3G-Vif complex. The molecular modeling study revealed that the loop7 motif of hA3G appears on the interfaces of both the hA3G-Vif complex and the hA3G dimer. Biochemical analysis provided evidence suggesting that binding of Vif to hA3G results in steric blocking of hA3G dimerization, implying that monomeric hA3G serves as a substrate for Vif-mediated degradation. Furthermore, we presented evidence for the important roles of the loop7 motif, especially the central residues within the region, in hA3G dimerization, hA3G--Vif interaction, Vif-mediated hA3G degradation as well as subcellular localization of hA3G. This work highlights a multiple-task interface formed by loop7 motif, which regulates biological function of hA3G, thus providing the feasibility of the strategy of blocking Vif-mediated A3G degradation by targeting the putative site around loop7.

A Novel Flow Diverter (Tubridge) for the Treatment of Recurrent Aneurysms: A Single-Center Experience

OBJECTIVE: The Tubridge flow diverter (FD) is a novel device aimed at reconstructing the parent artery and occluding complex aneurysms. Retreatment of recurrent aneurysms using the FD is challenging. We report our initial experience in the repair of aneurysm recurrence with the FD. MATERIALS AND METHODS: A database was reviewed prospectively, and 8 patients with 8 recurrent aneurysms (mean size, 16.7 mm) were identified. Four aneurysms had previously ruptured. The previous aneurysm treatment consisted of coiling in 1 aneurysm and single-stent-assisted coiling in 7 aneurysms. The procedural complications and clinical and angiographic outcomes were analyzed. RESULTS: Six aneurysms were treated by using a single Tubridge FD alone, while the remaining 2 were treated with FD + coiling. The immediate results of the 8 aneurysms were that they all showed incomplete occlusion. Neither major ischemic nor hemorrhagic complications occurred; however, 1 patient experienced a vasospasm. Follow-up angiographies were available for 7 aneurysms; the mean follow-up was 16.9 months (7–36 months). Five aneurysms were completely occluded, whereas 2 had a residual neck. Severe asymptomatic stenosis of 1 parent artery of a vertebral artery dissecting aneurysm was found. All visible branches covered by the FD were patent. All patients were clinically assessed as having attained a favorable outcome (modified Rankin Scale score ≤ 2) at discharge and follow-up. CONCLUSION: In selected patients, the Tubridge FD can provide a safe and efficient option for the retreatment of recurrent aneurysms. Nevertheless, attention should be paid to several technical points.

A cell-based high-throughput approach to identify inhibitors of influenza A virus

Influenza is one of the most common infections threatening public health worldwide and is caused by the influenza virus. Rapid emergence of drug resistance has led to an urgent need to develop new anti-influenza inhibitors. In this study we established a 293T cell line that constitutively synthesizes a virus-based negative strand RNA, which expresses Gaussia luciferase upon influenza A virus infection. Using this cell line, an assay was developed and optimized to search for inhibitors of influenza virus replication. Biochemical studies and statistical analyses presented herein demonstrate the sensitivity and reproducibility of the assay in a high-throughput format (Z' factor value>0.8). A pilot screening provides further evidence for validation of the assay. Taken together, this work provides a simple, convenient, and reliable HTS assay to identify compounds with anti-influenza activity.

Variation regularity of chemical constituents contained in Euphorbia ebracteolata before and after roasting with vinegar / 中国中药杂志

<p><b>OBJECTIVE</b>To study the variation regularity of chemical constituents contained in Euphorbia ebracteolata after vinegar processing.</p><p><b>METHOD</b>The colorimetric method was adopted for determining the variation of total lactone content in toxic constituents contained in E. ebracteolata decoction, with Kedde as the coloring reagent. The HPLC method was used for detecting the content variation of jolkinolide B and jolkinolide C, both were active constituents contained in E. ebracteolata decoction, before and after roasting with vinegar, in which Kromasil-ODS column (4.6 mm x 250 mm, 5 microm) was adopted, with the detection wavelength of 290 nm, column temperature at 25 degrees C , gradient elution with acetonitrile and water and the flow rate of 1.0 mL x min(-1).</p><p><b>RESULT</b>After roasting with vinegar, the total lactone content in E. ebracteolata was reduced from 0.60 to 0.45 mg x g(-1) , with active constituents, jolkinolide B and jolkinolide C, increased to varying degrees. The established chromatographic fingerprint contained risk information and could reflect the overall variation regularity of chemical constituents after roasting with vinegar.</p><p><b>CONCLUSION</b>The chemical constituents of E. ebracteolata show significantly changes, especially a reduced toxicity, after roasting with vinegar. The increase in its efficacy may be related to the variation of these constituents.</p>

GC-MS analysis on volatile components of wild Trogopterus faeces from Laishui county of Hebei province / 中国中药杂志

<p><b>OBJECTIVE</b>To analysis the volatile components of Trogopterus faeces.</p><p><b>METHOD</b>The volatile components of Trogopterus faeces was extracted by steam distillation and analyzed by GC-MS.</p><p><b>RESULT</b>Eighty-five volatile components were identified and the main components are cedrol (12.31%) and caryophyllene (7.5%).</p><p><b>CONCLUSION</b>The volatile components of the samples are originated from the ingredients or other chemical compositions with similar basic structure of the fodders.</p>

Analysis of ERG11 gene mutations in fluconazole-resistant Candida glabrata strains / 中华传染病杂志

Objective To study the mutations of ERG11 gene which encodes P450 lanosterol 14-α demethylase, and to explore the possible role of ERG11 gene in inducing fluconazole resistance in Candida glabrata. Methods ERG11 genes of 9 fluconazole-resistant Candida glabrata isolates and 10 fluconazole-sensitive Candida glabrata isolates were cloned into pUC57-T vector. The open reading frame of ERG11 gene were sequenced by two directional sequencing using universal primers. All sequences were compared with the published sequence. Results Ten kinds of synonymous point mutation were found. Neither missense mutation nor frame-shifting mutation was found. Among the 10 kinds of synonymous point mutation, 5 were found in both fluconazole-resistant and fluconazolesensitive Candida glabrata isolates, and 3 were only found in fluconazole-resistant isolates, 2 were only found in fluconazole-sensitive ones. The majority of the point mutations were located between 1320-2200 base pair of ERG11 gene. Conclusions There are ERG11 gene polymorphisms in clinical strains of Candida glabrata. ERG11 gene mutations are not found to be involved in the development of fluconazole resistance in Candida glabrata.

Determination of methyl nonyl ketone in volatile oil from herbs of Houttuynia cordata by GC-MS / 中国中药杂志

<p><b>OBJECTIVE</b>To develop a GC-MS method for the determination of methyl nonyl ketone in the volatile oil from the herbs of Houttuynia cordata.</p><p><b>METHOD</b>The sample was split in the 240 degrees C injection port, with 20:1 split ratio, and separated on a DB-5 (30 m x 0.25 mm, 0.25 microm film thickness) fused silica column with helium as the carrier gas. The temperature program was as follows: 100 degrees C for 2 min, the 5 degrees C x min(-1) to 150 degrees C, then 15 degrees C x min(-1) to 200 degrees C, and kept for 10 min. The MS transfer line temperature was set to 250 degrees C, the MS source temperature was set to 200 degrees C. The ionization mode was electron ionization (EI) and the selective ion monitor mode was used.</p><p><b>RESULT</b>A good linear relationship was constructed over the injection amount range of 5.5-110 ng of methyl nonyl ketone. The average recovery was 98.9%, and RSD was 2. 2%.</p><p><b>CONCLUSION</b>The developed method was sensitive, accurate, and can be used for the determination of methyl nonyl ketone in the volatile oil and for the quality control of H. cordata.</p>

Construction of a new vector for targeted gene disruption in Candida albicans / 中华传染病杂志

Objective To explore the feasibility of using Zeocin resistance as a new positive selection marker for the genetic manipulation of Candida albicans (C.albicans).Methods The susceptibility of C.albicans strain CAI4 to Zeocin was determined using a broth microdilution method in pH range from 6.0 to 8.0.The ZeoR expression cassette was amplified from plasmid pGAPZ-α-A by polymerase chain reaction (PCR).An integrated vector used to knockout AAF1 gene was constructed by gene splicing.Then the C.albicans CAI4 was transformed with this integrated plasmid containing the ZeoR expression cassette using lithium chloride method and the target AAF1 gene was replaced.The recombinant C.albicans was screened and the genome was extracted and amplified by PCR with two pairs of primers,then confirmed using genetic method.Results The C.albicans CAI4 was sensitive to Zeocin in pH range from 6.0 to 8.0.And the concentration of Zeoein(100 mg/L) at pH 7.0 was used to select recombined C.albicans.The recombined integrated vector was confirmed by DNA sequencing.The ZeoR expression cassette was detected using PCR method in recombined C.albicans.The AAF1 gene was replaced by ZeoR gene.Conclusion A new vector for targeted gene disruption in C.albicans is successfully constructed.

Construction of a genomic regulatory network based on gene expression profile of neonatal hypertrophic cardiomyocytes induced by phenylephrine / 中国病理生理杂志

AIM: To construct a genomic regulatory network based on gene expression profiling of hypertrophic cardiomyocytes induced by phenylephrine in neonatal rats. METHODS: Cultured neonatal hypertrophic cardiomyocytes were induced by phenylephedrine. The gene expression profiles of these cells were assessed by using a cDNA microarray, and the microarray data were further analyzed by Pathwaystudio and Agilent Literature Search software. RESULTS: A genes/proteins interaction network was constructed with 450 nodes and 592 edges by analyzing the gene expression in hypertrophic cardiomyocytes and literature mining. The network belongs to scale - free network by topological analysis, and 14 genes/proteins as key nodes, including PTPN11, TRAF6, HSPA8, VIM, RPS6KA3, PTHRP, GRB2 and PI3K, were predicted. Based on GO analysis, the genes/proteins associated with metabolism, signal transduction and cytoskeleton may play important roles in the process of cardiomyocytes hypertrophy induced by phenylephedrine in neonatal rat. CONCLUSION : The genomic regulatory network based on gene expression profiling and literature mining may provide integrated clue to elaborate hypotheses about the evolution of cardiomyocyte hypertrophy.

In Vitro Efficacy of Ravuconazole and Fluconazole Against Clinical and Environmental Isolates of Cryptococcus neoformans / 中华皮肤科杂志

0.05). Conclusion It is encouraging that ravuconazole is effective against clinical and environmental isolates of C. neoformans, whether the isolates were sensitive to fluconazole or not.

Assay for simultaneous detection of HIV p24 antigen and anti-HIV antibody / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To develop a rapid assay for simultaneous detection of HIV p24 antigen (Ag) and anti-HIV antibody (Ab).</p><p><b>METHODS</b>HIV-1 gp41 antigen and HIV-2 gp36 antigen were expressed by recombinant baculovirus insect system and purified by immunochromatography. p24 monoclonal antibody (mAb) was obtained from p24 hybridoma cell line. Purified antigen and mAb were dot blotted to nitrocellular membrane; 20 nm colloidal gold-anti-human IgG ab and p24 ab complex were used for this test. Previously detected 39 sera specimens were tested in this study to compare with the result of HIV test with commercial HIV test kit.</p><p><b>RESULTS</b>20 mg/L purified gp41 Ag and gp36 Ag were obtained from recombinant baculovirus-insect cell system; 1.5 mg/L p24 mAb was obtained from p24 mAb hybridoma cell line. Compared the test result of 39 sera with commercial HIV test kits, consistency rate was 100%.</p><p><b>CONCLUSIONS</b>The rapid assay for simultaneous detection of HIV p24 antigen and anti-HIV antibody provides a simple, sensitive and reliable test for HIV diagnosis.</p>

Study on relationship between efflux pumps and Candida albicans resistance mechanism / 中华传染病杂志

Objective To investigate whether efflux mechanism is involved in azole resistant Candida albicans strains isolated in China. Methods We compared rhodamine 6G uptake and glucose induced efflux between azole sensitive and azole resistant strains to elucidate resistance to azole due to efflux pump, then measured mRNA level of efflux gene cdr1 and cdr2 by reverse transcriptase polymerase chain reaction (RT PCR). Results Rhodamine 6G moved from extra into the intracellular compartment in both azole resistant and azole sensitive strains quickly when in glucose free incubation condition. However, efflux of rhodanmine 6G was enhanced significantly in azole resistant strains and decreased in azole sensitive strains while adding glucose in the media. The mRNA level of efflux gene cdr1 and cdr2 was higher in clinical azole resistant strains and fluconazole induced resistant strains than strains reverted from resistance to sensitivity. However, all such strains had higher efflux gene expression than azole sensitive strains. Conclusions Efflux mechanism is associated with the resistance to azole in Candida albicans strains isolated in China. Fluconazole can induce efflux pumps expression leading to resistance to azole but the resistance to azoles can be reversed after withdrawing fluconazole.

The point mutation of cytochrome P-450 lanosterol 14-? demethylase ERG11 gene in Fluconazole-resistant Candida albicans / 中华传染病杂志

Objective To investigate the point mutation of the open reading frame of cytochrome P-450 lanosterol 14-? demethylase gene ERG11. Methods In order to identify such alterations, the DNA was extracted by enzyme lysis methods and the PCR was performed. The PCR products were purified and cloned into PBS vector, then transformed into DH5? and sequenced. Results Twenty nine point mutations were identified in this 15 resistant isolates, most of which were different from susceptible strains. The mutations included 12 missense substitutions: F72L, D81G, D116E, K128T, Y132H, E266D, D294G, S361P, M374V, P386L, H400R, and Q474K, of which 6 had not been described previously (D294G, S361P, M374V, P386L, H400R, and Q474K). The other mutations included a frameshift, and 17 silent mutations. Conclusions The point mutation of resistant isolate is different from that of the susceptible isolate. It is suggested that the drug resistance is related with by the newly found alterations, including D294G, S361P, M374V, P386L, H400R, Q474K, and a frameshift.