Objective To study the effect of Liangxue Tuizi Formula(LXTZF)on reactive oxygen species(ROS)-mediated NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome activation in Henoch-Sch?nlein purpura(HSP)rats,and to explore its possible mechanism in the treatment of HSP.Methods Twenty-four rats were divided randomly into four groups:control,model,LXTZF,and compound glycyrrhizin(CG)groups.Except for the control group,a model of HSP was established in the other groups by heat drugs combined with egg albumin.After successful modeling,rats in the LXTZF group were given LXTZF solution 7.47 g/kg,rats in the CG group were given CG solution 13.5 mg/kg by gavage,and rats in the control and model groups were given normal saline solution by gavage once a day for 4 weeks.Samples were collected 8 hours after the last gavage.Skin histopathology changes were observed by hematoxylin and eosin(HE)staining.Serum interleukin(IL)-18 and IL-1βlevels were detected by enzyme-linked immunosorbent assay(ELISA).Changes in ROS levels in the skin were detected by immunofluorescence.Apoptosis-associated speckle-like protein(ASC),NLRP3,cysteinyl aspartate-specific protease-1(Caspase-1)mRNA and protein expression levels in rat skin were detected by real-time quantitative polymerase chain reaction(RT-PCR)and immunohistochemistry and Western blot,respectively.Results The skin pathology in the model group showed obvious inflammatory cell infiltration compared with the control group.Serum IL-18 and IL-1β levels were significantly increased(P＜0.05),skin ROS levels were significantly increased(P＜0.05),and skin ASC,NLRP3,Caspase-1 mRNA and protein expression levels were significantly increased(P＜0.05).Inflammatory cell infiltration in the skin tissues of rats was alleviated in the LXTZF and CG groups compared with the model group,while serum levels of IL-18 and IL-1β were significantly decreased(P＜0.05).ROS levels in the skin were significantly decreased(P＜0.05),and mRNA and protein levels of ASC,NLRP3,and Caspase-1 in the skin were significantly decreased(P＜0.05).Conclusions The mechanism of LXTZF in HSP may be related to the inhibition of ROS-mediated NLRP3 inflammasome activation.

Objective To study the regulatory effect of Qufeng Xiaodian Formula on serum differential metabolites of allergic purpura,and provide scientific basis for the diagnosis and treatment of allergic purpura in traditional Chinese medicine.Methods Sixty rats were randomly divided into a blank control group(referred to as the blank group),a model group,a compound glycyrrhizin group,a low-dose Qu Feng Xiao Dian Fang group,a medium dose Qu Feng Xiao Dian Fang group,and a high-dose Qu Feng Xiao Dian Fang group according to a random number table method,with 10 rats in each group.The model group was constructed by combining dried ginger,pepper,and long pepper with ovalbumin to create an allergic purpura rat model.After successful modeling,each treatment group was intervened with corresponding drugs for 4 weeks.After 4 weeks,serum was collected and non targeted metabolomics screening of serum differential metabolites was performed using ultra-high performance liquid chromatography quadrupole time-of-flight tandem mass spectrometer(UPLC-QTOF/MS).Subsequently,data extraction and multivariate statistical analysis will be conducted to identify potential metabolic pathways.Results Compared with the control group,there were 91 possible differential metabolites in the serum of the model group rats,corresponding to 20 metabolic pathways;Compared with the model group,there were a total of 43 possible differential metabolites in the serum of rats in the wind dispelling and disease eliminating group,corresponding to 15 metabolic pathways.Among them,there are a total of 12 metabolic pathways.Inflammatory metabolites such as arachidonic acid and ceramide can damage vascular endothelium.Ten biomarkers,including arachidonic acid and ceramide,were significantly abnormal in the serum of the model group rats compared to the normal group.The Qufeng Xiaodian formula can significantly reverse these metabolites and significantly enrich them in arachidonic acid metabolism and sphingolipid metabolism pathways.Conclusion Qufeng Xiaodian Formula has a certain regulatory effect on metabolites such as arachidonic acid and ceramide that affect vascular endothelial injury.

ObjectiveTo investigate the potential mechanism of Liangxue Tuizi Formula （凉血退紫方， LXTZF） in treating Henoch-Schönlein Purpura （HSP） by examining its regulatory effect on neutrophil extracellular trap （NETs） dysregulation via the rapidly accelerated fibrosarcoma kinase （RAF）/mitogen-activated protein kinase （MEK）/extracellular signal-regulated kinase （ERK） signaling pathway. MethodsSeventy Wistar rats were randomly allocated into a blank control group （n=14） and a modeling group （n=56）. Rats in the modelling group underwent an eight-week modelling period to establish HSP rat models with blood-heat syndrome via modified ovalbumin （OVA） induction method combined with oral administration of heat-property Chinese herbal medicine. Fifty successfully modeled rats were subsequently randomly divided into five groups （n=10 per group）， model group， compound glycyrrhizin group， LXTZF group， RAF inhibitor group， and LXTZF + RAF agonist group. Additionally， 10 rats were selected from the original blank control group for the final experiment. From the 11th week of modelling， rats in the blank control group and the model group received 1 ml/（100 g·d） ultrapure water via oral administration， in addition to 0.5 ml/（kg·d） 0.9% sodium chloride solution via intraperitoneal injection. The LXTZF group and the compound glycyrrhizin group received 7.5 g/（kg·d） LXTZF granule suspension via gavage， 13.5 mg/（kg·d） compound glycyrrhizin suspension via gavage， respectively. The RAF inhibitor group received 1 mg/（kg·d） GW5074 suspension via intraperitoneal injection and ultrapure water via oral administration； the LXTZF + RAF agonist group received 7.5 g/（kg·d） LXTZF granule suspension via gavage and 1 mg/（kg·d） paclitaxel suspension via intraperitoneal injection. All administrations were performed once daily for 4 weeks. After intervention， skin tissue histopathology was examined by hematoxylin and eosin （H&E） staining， immunoglobulin A （IgA） deposition was assessed via immunofluorescence， serum levels of neutrophil elastase （NE）， tumor necrosis factor-α （TNF-α）， and vascular cell adhesion molecule-1 （VCAM-1） were measured using enzyme-linked immunosorbent assay （ELISA）， serum myeloperoxidase （MPO） level was determined by a colorimetric assay； the mRNA expression levels of RAF， MEK， and ERK in skin tissue were detected by real-time quantitative polymerase chain reaction （RT-qPCR）； and the protein expression of RAF， MEK， ERK， as well as phosphorylated MEK （p-MEK） and phosphorylated ERK （p-ERK）， were analyzed by Western Blot. ResultsSkin tissue in the blank control group rats remained normal， whereas the model group exhibited neutrophil infiltration and haemorrhage with red blood cell rupture. In all drug intervention groups， neutrophil infiltration and haemorrhagic exudation reduced markedly， with LXTZF group demonstrating the most pronounced improvement. Compared with the blank control group， rats in the model group exhibited enhanced IgA fluorescence intensity in skin tissue， elevated serum levels of NE， MPO， TNF-α and VCAM-1， increased mRNA expression of RAF， MEK， ERK1 and ERK2， as well as heightened RAF protein levels and p-MEK/MEK and p-ERK/ERK ratios （P＜0.05）. Compared with the model group， the drug intervention groups exhibited reduced IgA fluorescence intensity in skin tissue， along with decreased serum levels of NE， MPO， TNF-α， and VCAM-1 （P＜0.05）. In LXTZF group and RAF inhibition groups， reduced mRNA expression of RAF， MEK， ERK1， and ERK2 was observed in rat skin tissue， alongside decreased RAF protein levels and reduced p-MEK/MEK and p-ERK/ERK ratios （P＜0.05）. Compared with LXTZF + RAF agonist group， the compound glycyrrhizin group， LXTZF group， and RAF inhibitior group exhibited reduced IgA fluorescence intensity in skin tissue， decreased serum NE， MPO， TNF-α， and VCAM-1 levels， and decreased MEK mRNA expression and p-MEK/MEK ratio （P＜0.05）. ConclusionThe potential mechanism by which LXTZF treats Henoch-Schönlein purpura with blood heat syndrome may involve blocking the RAF/MEK/ERK signaling pathway in skin tissue， and suppressing excessive formation of NETs， thereby reducing IgA deposition in dermal microvessels and attenuating systemic inflammatory responses.

Objective To study the effect of Liangxue Tuizi Formula(LXTZF)on reactive oxygen species(ROS)-mediated NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome activation in Henoch-Sch?nlein purpura(HSP)rats,and to explore its possible mechanism in the treatment of HSP.Methods Twenty-four rats were divided randomly into four groups:control,model,LXTZF,and compound glycyrrhizin(CG)groups.Except for the control group,a model of HSP was established in the other groups by heat drugs combined with egg albumin.After successful modeling,rats in the LXTZF group were given LXTZF solution 7.47 g/kg,rats in the CG group were given CG solution 13.5 mg/kg by gavage,and rats in the control and model groups were given normal saline solution by gavage once a day for 4 weeks.Samples were collected 8 hours after the last gavage.Skin histopathology changes were observed by hematoxylin and eosin(HE)staining.Serum interleukin(IL)-18 and IL-1βlevels were detected by enzyme-linked immunosorbent assay(ELISA).Changes in ROS levels in the skin were detected by immunofluorescence.Apoptosis-associated speckle-like protein(ASC),NLRP3,cysteinyl aspartate-specific protease-1(Caspase-1)mRNA and protein expression levels in rat skin were detected by real-time quantitative polymerase chain reaction(RT-PCR)and immunohistochemistry and Western blot,respectively.Results The skin pathology in the model group showed obvious inflammatory cell infiltration compared with the control group.Serum IL-18 and IL-1β levels were significantly increased(P＜0.05),skin ROS levels were significantly increased(P＜0.05),and skin ASC,NLRP3,Caspase-1 mRNA and protein expression levels were significantly increased(P＜0.05).Inflammatory cell infiltration in the skin tissues of rats was alleviated in the LXTZF and CG groups compared with the model group,while serum levels of IL-18 and IL-1β were significantly decreased(P＜0.05).ROS levels in the skin were significantly decreased(P＜0.05),and mRNA and protein levels of ASC,NLRP3,and Caspase-1 in the skin were significantly decreased(P＜0.05).Conclusions The mechanism of LXTZF in HSP may be related to the inhibition of ROS-mediated NLRP3 inflammasome activation.

Objective To study the regulatory effect of Qufeng Xiaodian Formula on serum differential metabolites of allergic purpura,and provide scientific basis for the diagnosis and treatment of allergic purpura in traditional Chinese medicine.Methods Sixty rats were randomly divided into a blank control group(referred to as the blank group),a model group,a compound glycyrrhizin group,a low-dose Qu Feng Xiao Dian Fang group,a medium dose Qu Feng Xiao Dian Fang group,and a high-dose Qu Feng Xiao Dian Fang group according to a random number table method,with 10 rats in each group.The model group was constructed by combining dried ginger,pepper,and long pepper with ovalbumin to create an allergic purpura rat model.After successful modeling,each treatment group was intervened with corresponding drugs for 4 weeks.After 4 weeks,serum was collected and non targeted metabolomics screening of serum differential metabolites was performed using ultra-high performance liquid chromatography quadrupole time-of-flight tandem mass spectrometer(UPLC-QTOF/MS).Subsequently,data extraction and multivariate statistical analysis will be conducted to identify potential metabolic pathways.Results Compared with the control group,there were 91 possible differential metabolites in the serum of the model group rats,corresponding to 20 metabolic pathways;Compared with the model group,there were a total of 43 possible differential metabolites in the serum of rats in the wind dispelling and disease eliminating group,corresponding to 15 metabolic pathways.Among them,there are a total of 12 metabolic pathways.Inflammatory metabolites such as arachidonic acid and ceramide can damage vascular endothelium.Ten biomarkers,including arachidonic acid and ceramide,were significantly abnormal in the serum of the model group rats compared to the normal group.The Qufeng Xiaodian formula can significantly reverse these metabolites and significantly enrich them in arachidonic acid metabolism and sphingolipid metabolism pathways.Conclusion Qufeng Xiaodian Formula has a certain regulatory effect on metabolites such as arachidonic acid and ceramide that affect vascular endothelial injury.

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.

OBJECTIVE:To study the effect of catalpol on the activity and apoptosis of osteoclasts (OC) in the osteoblasts (OB)-OC co-culture system and its mechanism. METHODS:OB and OC were isolated respectively from the SD rats of 1-3 days and 5-7 days old to establish OB-OC co-culture system. After treated with 0(blank control),0.05,0.5,5,50 and 100 mg/L catal-pol for 48,72 and 96 h,the number of bone absorption lacuna for OC was observed by inverted microscope to reflect OC activity. After treated with 0(blank control)and 0.05 mg/L catalpol for 48,72 and 96 h,the activity of tartrate resistant acid phosphatase (TRACP)in OC was detected,and the apoptosis rate of OC was calculated. After treated with 0(blank control)and 0.05 mg/L ca-talpol,mRNA expression of osteoprotegerin(OPG)in OB was detected. RESULTS:In OB-OC co-culture system,the number of bone absorption lacuna in 0.05-50 mg/L catalpol groups was significantly lower than blank control group(P<0.01),indicating ca-talpol could inhibit OC activity,especially 0.05 mg/L catapol. Compared with blank control,0.05 mg/L catapol lowered the activity of TRACP but increased the apoptosis rate of OC(P<0.05);mRNA expression of OPG was up-regulated in OB(P<0.01). CON-CLUSIONS:In OB-OC co-culture system,catalpol can inhibit the activity of OC and induce the apoptosis of OC,and its mecha-nism may be associated with the mRNA expression up-regulation of OPG in OB.

[ ABSTRACT ] AIM: To investigate the effect of catalpol on the activity of osteoblasts ( OB ) and osteoclasts ( OC) , and OB estrogen receptor ( ER) α/βmRNA expression in the OB-OC co-culture system.METHODS: OB and OC were isolated from the SD rats of 1 and 5 days old.In the OB-OC co-culture system, different concentrations of catalpol including low dosage (0.05 , 0.1, 0.5 and 1 mg/L), middle dosage (2, 5 and 10 mg/L), and high dosage (20, 50 and 100 mg/L) were added into the culture medium to detect the changes of OB proliferation by MTT assay.The catalpol at maximal dosage was added to OB section to detect the alkaline phosphatase ( ALP) activity of OB by pNPP method.The mRNA expression of ERα/βin the OB treated with catalpol in the co-culture system was detected by RT-PCR.The catalpol at maximal dosage was added to OC group to detect the activity of OC by microscopy and tartrate-resistantacid phosphatase ( TRAP) activity detection.RESULTS:In 0.05~2 mg/L catalpol groups, the proliferation of OB was significantly in-creased as compared with control group in the co-culture system, and it reached the maximum value when catalpol was at 0.05 mg/L, while in 5~100 mg/L catalpol groups, the proliferation of OB was not increased.The ALP activity of OB in 0.05 mg/L catalpol group was higher than that in control group.The catalpal at 0.05 mg/L promoted the mRNA expression of ERβin OB in the co-culture system, but did not increase the mRNA expression of ERαas compared with control group. Catalpol at 0.05 mg/L obviously inhibited the bone resorption and the TRAP activity in OC.CONCLUSION: Catalpol stimulates the proliferation and activity of OB, inhibits the bone resorption and activity of OC, and increases the mRNA ex-pression of ERβin OB in the OB-OC co-culture system, suggesting that high mRNA expression of ERβmay be the regula-tory pathway of catalpol in response to bone metabolism.

Objective To evaluate the clinical effect and safety of ureteroscopic pneumatic lithotripsy combined with transurethral resection of prostate(TURP)for treatment of benign prostatic hyperplasia(BPH)with bladder calculus.Methods 68 patients of BPH with bladder calculus treated by ureteroscopic pneumatic lithotripsy combined with transurethral resection of prostate were reviewed.Results Satisfactory effects were achieved in all these 68 cases without serious complications.Conclusion Ureteroscopic pneumatic lithotripsy combined with TURP is a minimally invasive,safe and effective method in the treatment of BPH and bladder calculus.

Objective To explore the correlation of PAI-1 and TNF-α and the pathophysiology of the metabolic syndrome (MS) and coronary heart disease,and explore the role of metformin in the MS.Methods Sixty cases of old patients with the MS were chosen.These patients were divided into two groups at random.One group interfered with living style and metformin,the other group only interfered with living style.The activity of PAI-1 was detected by chromogenic substrate method,and the level of TNF-α was detected by ELISA assay.Results (1) The levels of PAI-1 and TNF-α in the MS patients [(0.95 ± 0.05) AU/ml,(24.81 ± 3.87)ng/ml] were significantly higher than in normal old people[(0.66 ± 0.10)AU/ml,(10.76 ±2.00) ng/ml] (P ＜0.001) ;(2)The levels of PAI-1 and TNF-α in the MS patients with CHD [(0.96 ± 0.05) AU/ml,(26.12 ± 2.83) ng/ml] were significantly higher than those in the patients without CHD [(0.94 ± 0.03) AU/ml,(23.71 ± 4.27) ng/ml] (P ＜ 0.05) ;(3)The activity of PAI-1 and the level of TNF-α in the metformin group was decreased significantly [△ was (0.20 ± 0.17)AU/ml,(4.42 ± 0.85ng/ml),P ＜0.01],and metformin can improve the components of the MS.Conclusions The old patients with MS is prone to develop cardiac vascular disease.PAI-1 and TNF-α participate in pathophysiology of the MS and its complication.Metformin can inhibit the expression of PAI-1 and TNF-α to suppress the components of the MS,and block the complication of the MS.