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COVID-19 is caused by a novel coronavirus-severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), which has being spreading around the world, posing a serious threat to human health and lives. Neutralizing antibodies and small molecule inhibitors for virus replication cycle are the main antiviral treatment for novel coronavirus recommended in China. To further promote the rational use of antiviral therapy in clinical practice, the National Center for Infectious Diseases (Beijing Ditan Hospital Capital Medical University and the First Affiliated Hospital, Zhejiang University School of Medicine) invited experts in fields of infectious diseases, respiratory and intensive care to develop an Expert Consensus on Antiviral Therapy of COVID-19 based on the Diagnosis and Treatment Guideline for COVID-19 ( trial version 10) and experiences in the diagnosis and treatment of COVID-19 in China. The consensus is concise, practical and highly operable, hopefully it would improve the understanding of antiviral therapy for clinicians and provide suggestions for standardized medication in treatment of COVID-19.
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OBJECTIVE:To optimize the purification technology of total flavones from Ampelopsis grossedentala with macro-porous resins. METHODS:4 kinds of macroporous resins for the purification of total flavones from A. grossedentala were screened by using drug-loading amount,desorption rate,recovery and purification rate as indicators. Single factor test and central composite design-response surface methodology were used to optimized eluant mass fraction,adsorption time,flow rate of eluant,eluant pH and other factors of purification technology,and validation test was also conducted. RESULTS:D-101 macroporous resin was the best. The optimal condition was as follows as the concentration of sample solution 2 mg(by extract weight)/ml,the volume of sam-ple solution 1.1 BV,ethanol 86.0%,adsorption time 36.7 min,flow rate of eluant 3.81 BV/h,pH 7. In validation test,mass frac-tion of total flavones increased from 66.83% to 85.00% in validation test(RSD=0.15%,n=3),and were close to predicted val-ue(85.08%). CONCLUSIONS:Central composite design-response surface methodology is feasible and stable for the optimization of purification technology of total flavones from A. grossedentala with macroporous resins.
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Objective To study the protective effect of erythropoietin (EPO) on brain tissue with cardiac arrest-cardiopulmonary resuscitation (CA-CPR) and its mechanism.Methods 120 male Sprague-Dawley (SD) rats were randomly divided into three groups (each n =40),namely:sham group,routine chest compression group,and conventional chest compression + EPO group (EPO group).The rats in each group were subdivided into CA and 6,12,24,48 hours after restoration of spontaneous circulation (ROSC) five subgroups (each n =8).The model of CA was reproduced according to the Hendrickx classical asphyxia method followed by routine chest compression,and the rats in sham group only underwent anesthesia,tracheostomy intubation and venous-puncture without asphyxia and CPR.The rats in EPO group were given the routine chest compression + EPO 5 kU/kg (2 mL/kg) after CA.Blood sample was collected at different time points of intervention for the determination the content of serum S100 β protein by enzyme linked immunosorbent assay (ELISA).All the rats were sacrificed at the corresponding time points,and the hippocampus was harvested for the calculation of the number of S100 β protein positive cells,and to examine the pathological changes and their scores at 24 hours after ROSC by light microscopy.Results With prolongation of ROSC time,the serum levels of S100 β protein (μg/L) in the routine chose compression group and the EPO group were significantly elevated,peaking at 24 hours (compared with CA:305.7 ± 29.2 vs.44.4 ± 6.2 in routine chest compression group,and 276.7±28.9 vs.44.7±5.6 in the EPO group,both P ＜ 0.05),followed by a fall.The levels of S100β protein at each time point after ROSC in EPO group were significanthy lower than those of the routine chest compression group (83.2 ± 7.5 vs.114.3 ± 15.3 at 6 hours,123.9 ± 20.2 vs.184.9 ± 22.2 at 12 hours,276.7 ± 28.9 vs.305.7 ± 29.2 at 24 hours,256.3 ± 26.6 vs.283.2 ± 23.6 at 48 hours,all P ＜ 0.05).With the prolongation of ROSC time,the S100 β protein positive cell number in brain (cells/HP) in the routine chest compression group and the EPO group was significantly increased,peaking at 24 hours (compared with CA:14.3±2.2 vs.6.7±0.7 in the routine chest compression group,11.3± 1.3 vs.6.8±0.9 in the EPO group,both P ＜ 0.05),then it began to fall.The S100 β protein positive cell number in brain at each time point after ROSC in the EPO group was significantly lower than that of the routine chest compression group (7.0±0.9 vs.7.9± 1.9 at6 hours,8.4± 1.1 vs.10.2±2.2 at 12 hours,11.3± 1.3 vs.14.3±2.2 at24 hours,8.3±0.8 vs.10.8±2.0 at48 hours,all P ＜ 0.05).Under the light microscope,a serious brain cortex injury was found after reproduction of the model,and the degree of injury was reduced after EPO intervention.The pathological score at 24 hours after ROSC in EPO group was lower than that of routine chest compression group (3.83±0.73 vs.4.17±0.75,P ＜ 0.05).Conclusions The S100β protein level in serum and brain tissue was increased early in asphyxia CA-CPR rats.EPO intervention can reduce the expression of S100 protein and reduce the degree of brain injury.
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Objective To study the regulation of aquaporin-1(AQP-1)changes in the heart of septic rats, compare the correlations of the AQP-1 with myocardial cytokines tumor necrosis factor-α(TNF-α),interleukin-6 (IL-6),and myocardial tissue water content,and to investigate the dexmedetomidine protective effect on myocardia in septic rats and its possible mechanism. Methods According to the random number table methods,90 male Sprague-Dawley(SD)rats were divided into sham operation group,sepsis model group and dexmedetomidine group, 30 rats in each group. The rat sepsis model was established by cecal ligation and puncture(CLP). In the sham operation group,the animal abdomen was only opened and closed without CLP. Half hour before operation in dexmedetomidine group,dexmedetomidine 1μg/kg(2μg/mL)was injected into the vein,while in the model and sham groups,saline 5 mL/kg was subcutaneously injected into the rat after the operation. At 2,12,24,48,72 hours after operation,6 rats were sacrificed and their hearts removed at one time point in a group. Enzyme linked immunosorbent assay(ELISA)was used to detect the content of AQP-1 and the levels of the TNF-α,IL-6 in the myocardial tissue homogenate at all time points,the myocardial tissue water content was detected by dry wet weight,and the correlations between AQP-1 and TNF-α,IL-6 and between AQP-1 and myocardial tissue water content were compared. Results From 2 hours after operation,the levels of the AQP-1,TNF-αand IL-6 in model group were significantly higher than those in the sham operation group;with prolongation of time,the level of AQP-1 and myocardial tissue water content were decreased, but the levels of TNF-α and IL-6 were persistently increased. From 2 hours after operation in dexmedetomidine group,all the above indexes except myocardial tissue water content at 72 hours after operation were significantly lower than those in the model group〔AQP-1(ng/g):9.29±0.15 vs. 9.73±0.26,TNF-α(pg/g):109.47±8.41 vs. 128.13±7.36,IL-6(pg/g):232.95±20.56 vs. 279.71±22.24,myocardial tissue water content:(74.82±6.37)%vs.(75.62±6.39)%,all P<0.05〕,but still higher than those of the sham operation group. The correlation analyses for the septic group showed that the change of AQP-1 was positively correlated to the myocardial water content in early stage(r=0.418,P=0.001)and later stage(r=0.235,P=0.022),and the changes of the AQP-1 in early stage (at post-operative 2 hours)were positively correlated to the concentration changes of the cytokines TNF-α(r=0.235,P=0.021)and IL-6(r=0.345,P=0.003),but in the later stage(at post-operative 72 hours)were negatively correlated with the changes of TNF-α(r=-0.408,P=0.037)and IL-6(r=-0.276,P=0.002). Conclusions In the early stage of septic rats,there is obvious myocardial injury,resulting in the over expression of AQP-1 and the occurrence of myocardial edema,dexmedetomidine can play a role in myocardial protection in such rats and its mechanism is possibly related to the reduction of the expression of AQP-1 and the levels of inflammatory cytokines, and in turn the alleviation of myocardial cell edema.
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Objective To investigate the protective effects of ulinastatin on the hearts of rats with anoxia-induced cardiac arrest-cardiopulmonary resuscitation(CA-CPR) and the mechanism of improving cardiac dysfunction .Methods Twenty male Sprague Dawley(SD) rats were randomly divided into three experimental groups :sham operation group (group A ,n= 8 ,only anesthesia , tracheotomy tube and vascular puncture) ,control group(group B ,n= 6 ,normal saline 4 mL · kg -1 injected via vein) ,Ulinastatin treatment group(group C ,n=6 ulinastatin 50 000 U/kg+normal saline 3 mL · kg -1 injected via vein);Factors including mean arte-rial pressure(MAP) ,left ventricular end diastolic pressure(LVEDP) ,the maximum rising and falling rates of left ventricular deep pressure(± LVdp .dt-1max) ,brain natriuretic peptide(BNP) ,cardiac troponin T(cTNT) ,IL-12 and TNF-αwere observed at setting time before and after cardiopulmonary resuscitation in rats .Results Compared with those of the group A and before CA-CPR ,the concentrations of IL-12、cTNT、TNF-α、BNP、and LVEDP increased(P<0 .01)while ± LVdp .dt-1max decreased(P<0 .01) at 6 h after CA-CPR in group B ,C .Compared with those of group B ,the concentrations of IL-12、CTNT、TNF-α、BNP and LVEDP of 6 h after CA-CPR in group C were lower and ± LVdp .dt-1max was higher(P<0 .01) ,The concentrations of MAP of 6 h after CA-CPR in group B was lower Compared with that of group A ,C and before CA-CPR(P<0 .01) .Conclusion Ulinastatin can improve cardiac dysfunction by depressing mediators of inflammation and reducing myocardial injury .
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Objective To investigate the effects of Xuebijing injection on the transcription factors GATA-3 and T-bet of rats after cardiopulmonary resuscitation (CPR). Method The rat models of CPR was made by using asphyxia method. Thirty SD rats were randomly (random number) divided into three groups at random: sham operation rats (B group), conventional CPR rats (C group) and Xuebijing (4 mL/kg) treated rats (D group). The levels of serum IL-12, IL-4, TNF-α and IFN-γ were measured by using ELISA. The expressions of T-bet and GATA-3 mRNA in serum were detected by RT-PCR. The analysis of variance was used to compare the means of different groups including t-test and Wilcoxon test. Results Compared with group B, the levels of serum IL-12, TNF-α and IFN-γ in group C and group D were significantly elevated after CPR for 6 hours (P<0.01). In group C, the expression of GATA-3 mRNA and GATA-3/T-bet decreased (both P < 0.05), while the expression of T-bet mRNA increased (P<0.01) after CPR for 6 hours. In group D, the expressions of GATA-3 mRNA and T-bet mRNA as well as GATA-3/T-bet increased after CPR for 6 hours. The levels of IL-12, IFN-γ and TNF-α in group D were lower than those in groupC (P<0.01). Compared with group C, the expression of GATA-3 mRNA and GATA-3/T-bet were significantly elevated and the expression of T-bet mRNA decreased in group D. ConclusionsThe transcription factors GATA-3 and T-bet may fail to get balance after CPR. The Xiebijing injectio can modulate the balance between GATA-3 and T-bet, and the levels of IL-12,IFN-γ and TNF-α.
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OBJECTIVE: To investigate the effect of Xuebijing injection on the cardiac function and calcium ions of cardiocyte in rats after anoxia-induced cardiac arrest-cardiopulmonary resuscitation(CA-CPR).METHODS: CA-CPR model was induced in rats and then the model rats were randomized to 4 groups,i.e.sham group,model group,Xuebijing injection high dose group(4 mL?kg-1),and Xuebijing injection lowdose group(Xuebijing injection 2 mL?kg-1).Mean arterial pressure(MAP),?LVdp/dt max,average fluorescence intensity of calcium and pathological changes of cardiocytes were observed at 0 and 6 hours after resuscitation,respectively.RESULTS: Compared with model group,Xuebijing ingection high dose group at 6 h showed significantly increased MAP(P