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OBJECTIVE@#Anemoside B4 (AB4), the most abundant triterpenoidal saponin isolated from Pulsatilla chinensis, inhibited influenza virus FM1 or Klebsiella pneumoniae-induced pneumonia. However, the anti-SARS-CoV-2 effect of AB4 has not been unraveled. Therefore, this study aimed to determine the antiviral activity and potential mechanism of AB4 in inhibiting human coronavirus SARS-CoV-2 in vivo and in vitro.@*METHODS@#The cytotoxicity of AB4 was evaluated using the Cell Counting Kit-8 (CCK8) assay. SARS-CoV-2 infected HEK293T, HPAEpiC, and Vero E6 cells were used for in vitro assays. The antiviral effect of AB4 in vivo was evaluated by SARS-CoV-2-infected hACE2-IRES-luc transgenic mouse model. Furthermore, label-free quantitative proteomics and bioinformatic analysis were performed to explore the potential antiviral mechanism of action of AB4. Type I IFN signaling-associated proteins were assessed using Western blotting or immumohistochemical staining.@*RESULTS@#The data showed that AB4 reduced the propagation of SARS-CoV-2 along with the decreased Nucleocapsid protein (N), Spike protein (S), and 3C-like protease (3CLpro) in HEK293T cells. In vivo antiviral activity data revealed that AB4 inhibited viral replication and relieved pneumonia in a SARS-CoV-2 infected mouse model. We further disclosed that the antiviral activity of AB4 was associated with the enhanced interferon (IFN)-β response via the activation of retinoic acid-inducible gene I (RIG-1) like receptor (RLP) pathways. Additionally, label-free quantitative proteomic analyses discovered that 17 proteins were significantly altered by AB4 in the SARS-CoV-2 coronavirus infections cells. These proteins mainly clustered in RNA metabolism.@*CONCLUSION@#Our results indicated that AB4 inhibited SARS-CoV-2 replication through the RLR pathways and moderated the RNA metabolism, suggesting that it would be a potential lead compound for the development of anti-SARS-CoV-2 drugs.
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Objective:To investigate plasma levels of fibrinopeptide A(FPA)and platelet alpha-granule membrane glycoprotein 140(GMP-140), and to analyze the changes of coagulation activity in elderly patients with primary microvascular angina(PMVA).Methods:In this prospective case-control study, a total of 45 elderly patients(aged 60-80 years, 27 males and 18 females)admitted to our hospital from Jan.2019 to Dec.2020 were diagnosed as PMVA(as microvascular angina group)by clinical symptoms, electrocardiogram, dynamic electrocardiogram and coronary angiography.Forty-three age-and sex-matched elderly subjects who took a medical check-up and had no primary microvascular angina were included as the control group.Plasma levels of FPA, GMP-140 and high-sensitivity C-reactive protein were determined.The levels of systolic blood pressure, diastolic blood pressure, platelet, fasting blood glucose, triglyceride, total cholesterol, high-density lipoprotein and low-density lipoprotein, uric acid, renal function and liver function were detected.Results:The FPA level was higher in the PMVA group than in the control group[(430.32±364.05)μg/L vs.(263.73± 118.29)μg/L, t=2.913, P<0.01]. There was no difference in GMP-140 level between the PMVA group and the control group[(5.78±3.92)μg/L vs.(6.95±1.91)μg/L, t=-1.790, P>0.05]. The high-sensitivity C-reactive protein level was higher in the PMVA group than in the control group[(3.33±5.70)mg/L vs.(0.81±0.86)mg/L, t=2.927, P<0.01]. Conclusions:Compared with the control group, the FPA level is significantly increased with elevated coagulation activity in patients with primary microvascular angina.The increased inflammatory state in the PMVA group might promote the occurrence and development of microvascular angina, but whether the inflammatory state promotes the enhancement of coagulation activity needs further study to confirm.
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A series of 2-(((5-akly/aryl-1-pyrazol-3-yl)methyl)thio)-5-alkyl-6-(cyclohexylmethyl)-pyrimidin-4(3)-ones were synthesized and their anti-HIV-1 activities were evaluated. Most of these compounds were highly active against wild-type (WT) HIV-1 strain (IIIB) with EC values in the range of 0.0038-0.4759 μmol/L. Among those compounds, had an EC value of 3.8 nmol/L and SI (selectivity index) of up to 25,468 indicating excellent activity against WT HIV-1. anti-HIV-1 activity and resistance profile studies suggested that compounds and displayed potential anti-HIV-1 activity against laboratory adapted strains and primary isolated strains including different subtypes and tropism strains (ECs range from 4.3 to 63.6 nmol/L and 18.9-219.3 nmol/L, respectively). On the other hand, it was observed that those two compounds were less effective with EC values of 2.77 and 4.87 μmol/L for HIV-1A (K103N + Y181C). The activity against reverse transcriptase (RT) was also evaluated for those compounds. Both and obtained sub-micromolar IC values showing their potential in RT inhibition. The pharmacokinetics examination in rats indicated that compound has acceptable pharmacokinetic properties and bioavailability. Preliminary structure-activity relationships and molecular modeling studies were also discussed.
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Objective To investigate the clinical significance of fraction exhaled nitric oxide (FeNO) in the diagnosis and monitoring of childhood bronchial asthma (asthma). Methods The FeNO levels of 204 children with acute asthma attack, 148 children with asthma clinical remission, 107 children with cough variant asthma (CVA) and 250 children with pneumonia from March 2016 to March 2017 were retrospectively analyzed and compared. Results The FeNO levels of acute asthma attack, asthma clinical remission, CVA and pneumonia were 18 (10, 37), 16 (10, 38), 18 (9, 31) and 13 (8, 20) nmol/L, and there was statistical difference (P<0.05). The FeNO levels of acute asthma attack, asthma clinical remission and CVA were significantly higher those of pneumonia, and there were statistical differences (P<0.05). There was no statistical difference in the FeNO levels among acute asthma attack, asthma clinical remission and CVA (P>0.05). Conclusions FeNO has clinical significance in the diagnosis of asthma in children, and its clinical significance in monitoring asthma and the prediction of acute attack needs further observations.
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Aim To establish and optimize the VSVG/HIV-1NL4-3 Luc pseudovirus model for anti-HIV drugs screening. Methods The infectivity of VSVG/HIV-1 NL4-3 Luc in 4 different cell lines was investigated according to the method of the lucifer-ase activity analysis system of Promega company. 3 different ex-perimental settings were used to detect the activities of approved anti-HIV drugs to confirm the feasibility and effectiveness of the system. Finally, some potential compounds were screened for their anti-HIV activities, and their antiviral activities against the pseudovirus were compared with HIV-1ⅢB . Results The pseud-ovirus showed the strongest replication ability in CRFK cells, and a clear dose-effect relationship was found between the report gene expression level and the virus quantity. Comparing the EC50 of different positive inhibitors against VSVG/HIV-1 NL4-3 Luc on 3 kinds of experimental conditions, 3rd scheme is the best. Finally, the system was used to screen compounds, the EC50 s a-gainst pseudovirus were similar to those in HIV-1ⅢB . Conclusion An optimized VSVG/HIV-1 NL4-3 Luc anti-HIV screening sys-tem has been successfully developed.
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Objective To screen the non-nucleoside compounds against HIV-1 reverse transcriptase by molecular modeling and bioactivity assay. Methods Surflex-Dock module of Tripos SYBYL software was used to simulate the binding pattern of 22 000 compounds in SPECS database with the active pocket of HIV-1 reverse transcriptase. Based on the simulation results, the interaction mode between the above compounds and the crystal structure of HIV-1 reverse transcriptase was analyzed. The compounds with higher docking scores and better binding pattern were determined by anti-HIV-1 ac tivities test in vitro. Results The virtual screening results showed that the docking conformation of 1- (4-fluorophenyl) -3- [2- (1H-indol-3-yl) ethyl] thiourea was similar to the embedded ligand in Rilpivirine crystal structure. 1- ( 4-fluorophenyl) -3- [ 2- ( 1H-indol-3-yl) ethyl] thiourea was held together with the key residue Lys101 in docking pocket of HIV-1 reverse transcriptase by hydrogen bonds, and hadπ-πstacking action together with the conservative residue Trp229 and the aromatic residue Tyr181 respectively. The bioassay in vitro results showed that when the proliferation rate of C8166 lymphocyte syncytium infected by HIV-1ⅢB arrived 50% ( EC50) , the concentration of 1- ( 4-fluorophenyl) -3- [ 2- ( 1H-indol-3-yl) ethyl] thiourea was 5.45μg/mL. Conclusion Molecule docking technology is an effective approach to reducing the screening of candidate compounds with micromolecular activity, and can be used to predict the interaction mode between the compound and the target receptor. In the study, active compound 1- (4-fluorophenyl) -3- [2- (1H-indol-3-yl) ethyl] thiourea has been screened out by molecule docking technology.
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Objective To explore the reference values of peak expiratory flow (PEF) for 4.0-14.0 years old children in Chengdu.Methods From November 2011 to May 2012,a total of 446 healthy children were enrolled in this study.PEF was measured using expiratory peak velocimeter and lung function meter.Children's age,sex,height and weight were recorded.Impact factors for PEF were investigated by multiple stepwise regression analysis.Results The PEF detected by expiratory peak velocimeter and lung function meter had a good correlation (r =0.920,P ＜ 0.01).Boys own higher peak expiratory flow rate than girls.The level of PEF had the best correlation with height,next was weight and age.The level of PEF increased with the increase of height,age.Some children PEF value was less than 80％ lower limits of lower limits of normal.The PEF could be obtained from height and age.Conclusions Males and females PEF can be calculated according to children's height.Set up the PEF equation about the expiratory peak velocimeter and lung function meter.
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Objective To investigate the effect of different etiology on the serum level of carbohydrate antigen 125 (CA125) in elderly patients with chronic congestive heart failure (CHF), and to assess any correlation of CA125 with serum level of B-type natriuretic peptide (BNP). Methods The 155 aged patients with New York Heart Association (NYHA) class Ⅲ or Ⅳ were enrolled and grouped into four reasons of hypertension, coronary heart disease (CHD), cardiomyopathy and other reasons, and 25 healthy old persons as control.CA125 and BNP levels were measured by automatic chemiluminescent immunoassay and enzyme immunoradiometric assay, respectively. Results CA125 level in patients with CHF was (83.4±6.6)U/L for hypertension, (36.8±1.4)U/L for CHD, (38.1±1.6)U/L for cardiomyopathy and (38.4±1.4)U/L for other reasons, which significantly higher than for healthy controls [(14.3±1.15) U/L, P＜0.05].Especially, CA125 level in hypertension group was notable higher than in other reasons of group (P＜0.05), and was positively related to BNP level (r=0.67,P＜0.05). Conclusions Serum CA125 level is a predictor for clinical pathogen of CHF.Therefore, it may be a useful additional marker for the evaluation of clinical treatment of these patients
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Objective To evaluate the effectiveness of noninvasive positive pressure ventilation(NPPV) as a weaning strategy in patients with acute respiratory failure after failure to wean from invasive positive pressure ventilation(IPPV). Method A prospective randomized and controlled clinical trial of weaning of IPPV was carried out in patients mechanically ventilated in mode of IPPV for more than 48 hours with failure in a spontaneous breathing trial(SBT: PSV 6 cmH_2O). Patients with contraindications to NPPV were excluded. After failure the SBT, patients were randomly divided(random number) in two groups. Patients in NPPV group were extubated after being ventilated with high pressure support for 30 minutes and then placed on NPPV. Patients in IPPV group were weaned following conventional procedure. Arterial blood gases, maximal inspiratory pressure, respiratory rate,tidal volume, rapid shallow breathing index, heart rate, arterial blood pressure, and peripheral oxygen saturation were measured before and after failing the SBT. The rate of complications, including pneumonia and tracheotomy duration mechanical ventilation, days of hospital stay and outcome were observed. Findings of the two groups were vompared using the Student t test and the chi-square test. Results The percentage of complications in the NPPV group was lower(22.9% versus 72.2%, P <0.01) ,with lower incidences of pneumonia(6.1%,36.1%; P <0.01) and tracheotomy. Compared between the two groups, days of ICU stay( 14.16(3.45) d vs. 22.57( 7.71 ) d; P <0.01) and total days of mechanical ventilation(14.88±3.76 days vs. 20.68± 2.79 days, P <0.01) of NPPV group are shorter than IPPV group. Conclusions NPPV is a good alternative to the mechanically venti-lated patients who fail in initial weaning attempts. The key to successful NPPV weaning is the proper selection of weaning candidates and using NPPV as soon as possible after extubation.
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Objective To explotr clinical application of impulse oscillation system(IOS)determining bronchodilator reversibility test and the impact of age factor on positive values of bronchoddilator reversibility test in children.Methods Two hundred and fifteen patients during asthma attack were recruited randomly for this study.They were divided into three groups according to their age(4-7 year-old group,8-11 year-old group and 12-16 year-old group).Mensurated MEFV and IOS lung function.Carried on the bronchial dilation test with the results.Forced expiratory volume in first second(FEV1),the sensitivity and specificity of IOS parameters,total respiratory impedance(Zrs),respiratory resistance at 5 Hz(R5)and 20 Hz(R20),reactance at 5 Hz(X5)and resonant frequency(Fres)for diagnosing asthma at different improvement levels after inhalation of bronchedilator were calculated respectively through using FEV1 as a "sold standard"in the different age groups.Results After inhalation of bronchodilator,Fres,Zrs,R5,X5 and R20 of IOS were improved significantly.The improvement of X5 was the most obvious,however the decline of R20 was the lowest.The improving rate of Fres,Zrs,R5 had very significant difference in the three age groups(P＜0.01).It displayed that the age factor was very important impact factor on the improving rate of Fres,Zrs,R5,however the improving rate of X5 was on difference in the three age groups(P＞0.05).In the three age groups,there were significantly positive correlations between X5,R5,Zrs and FEV1(P＜0.01).there wash't correlations between R20 and FEV1 in the 4-7 year-old group and the 12-16 year-old group(P＞0.05).Furthermore,when an increase of FEV1 equaling to or greater than 15% was used as reference for positive bronchial reversibility,the improve rate of Fres,Zrs,R5,and X5 equaled to or greater than 13%,23%,17%,31%,respectively in 4-7 year-old group,18%,26%,25%,32%,respectively in 8-11 year-old group,25%,28%,28%,36%,respectively in 12-16 year-old group. Conclusions IOS can be used to assess bronchodilator reversibility test in children,and age factors should be paid attention.
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AIM:This study was performed to investigate the feasibility and efficiency of exogenous mesenchymal stem cells(MSCs) transplantation on post-infarction ventricular remodeling and heart function in rats and compare the effects between adult rat MSCs and neonate rat MSCs transplantation.METHODS:1-2 hours after left coronary artery ligation,MSCs cultured in ex vivo,marked with BrdU,were injected directly into the border of infarcts in exogenous rats.6 weeks after transplantation,rat' heart function,ventricular remodeling and pathological results were measured.RESULTS:MSCs transplantation decreased LV end-diastolic diameter and end-systolic diameter,limited LV chamber dilatation and reduced collagen content significantly.The numbers of blood vessels and cardiomyocytes were increased.BrdU-labelled MSCs with oval nucleus were widely distributed.There were no significant difference between adult rat MSCs and neonate rat MSCs transplanted groups.CONCLUSION:MSCs can survive and home in exogenous host infarct hearts without addition of any immunosuppressant.MSCs transplantation has benificial effects on remodeling processes and contributes to improvement of cardiac function,which may be related with the reduction of the amount of the collagen,promotion of myogenesis and angiogenesis.