A throat swab sample from a pediatric case in Tongzhou District, Beijing was identified as enterovirus; the patient was a 1-year-and-8-month-old male sporadic case. Whole-genome sequencing revealed a viral genome length of 7 436 bp. BLAST alignment confirmed the serotype as EV-D68. Phylogenetic analysis of the whole genome indicated that this strain belongs to the B3 clade, showing closer genetic proximity to the 2018 Shanghai strain MW697453 with 99.53% whole-genome nucleotide homology. Genetic and amino acid variation analysis demonstrated that the B3 subclade to which this strain belongs exhibits a nucleotide deletion at positions 718–726, differing from deletion sites observed in other B3 clade strains. A key neuropathogenic amino acid site, T650A, was found to have undergone mutation. Recombination analysis confirmed no cross-clade recombination events in this strain. This study conducted genetic characterization of the strain's evolutionary relationship with EV-D68 strains from different regions and years in China, providing data support for formulating prevention and control measures against EV-D68 infection.

Objective To investigate the clinical efficacy of Guhong Injection for the treatment of patients with coronary microvascular dysfunction(CMD)of qi stagnation and blood stasis syndrome.Methods Sixty cases of patients with CMD of qi stagnation and blood stasis syndrome who were admitted to Guangdong Provincial Hospital of Chinese Medicine from June 2021 to August 2022 were randomly divided into the control group and the trial group according to the random number table method,with 30 cases in each group.The control group was treated with conventional western medicine,and the trial group was treated with Guhong Injection on the basis of treatment for the control group.The course of treatment for the two groups covered 10 days.The changes in traditional Chinese medicine(TCM)syndrome scores,and levels of lipid indicators,serum inflammatory factors and endothelial factors in the two groups were observed before and after treatment.After treatment,the clinical efficacy of the two groups was evaluated.Results(1)During the trial,three cases in the control group and two cases in the trial group fell off and a total of 55 cases were finally included in the statistical analysis of efficacy,including 27 cases in the control group and 28 cases in the trial group.(2)After 10 days of treatment,the total effective rate of the trial group was 89.29%(25/28),and that of the control group was 40.74%(11/27),and the intergroup comparison(tested by chi-square test)showed that the therapeutic efficacy of the trial group was significantly superior to that of the control group(P<0.01).(3)After treatment,the TCM syndrome scores of patients in both groups were decreased compared with those before treatment(P<0.05),and the decrease in the trial group was significantly superior to that in the control group(P<0.01).(4)After treatment,the levels of lipid indicators triglyceride(TG),total cholesterol(TC),and low-density lipoprotein cholesterol(LDL-C)in the two groups of patients were decreased compared with those before treatment(P<0.05),and the level of high-density lipoprotein cholesterol(HDL-C)was increased compared with that before treatment(P<0.05).The intergroup comparison showed that the decrease of TG,TC,and LDL-C levels as well as the increase of HDL-C level in the trial group was significantly superior to that in the control group(P<0.05 or P<0.01).(5)After treatment,the serum levels of inflammatory factors high-sensitivity C-reactive protein(hs-CRP),interleukin 6(IL-6)and tumor necrosis factor α(TNF-α)in the two groups of patients were decreased compared with those before treatment(P<0.05),and the decrease in the trial group was significantly superior to that in the control group(P<0.01).(6)After treatment,the serum level of endothelial factor nitric oxide(NO)in the two groups of patients was increased(P<0.05)and the serum endothelin 1(ET-1)level was decreased compared with that before treatment(P<0.05),and the increase of serum NO level,as well as the decrease of serum ET-1 level in the trial group was significantly superior to that in the control group(P<0.05 or P<0.01).Conclusion Based on the conventional treatment in western medicine,the application of Guhong Injection in the treatment of patients with CMD of qi stagnation and blood stasis syndrome exerts remarkable efficacy,which can effectively alleviate the symptoms,regulate the levels of blood lipids,reduce the inflammatory response,and improve the endothelial function.

AIM:To investigate the effect of intrinsic specific transforming growth factor-β(TGF-β)signaling on regeneration and repair of myofibers in acute skeletal myositismice model induced by cardiotoxin(CTX).METHODS:One hundred and eighty-six wild C57BL/6 mice and one hundred and thirty-eight mice with conditional knockout of TGF-β receptor 2(TGF-βr2)in myofibers(SM TGF-βr2-/-mice)were selected.CTX injection to anterior tibial muscle(TA)in-duced acute myoinjury in mice.Some SM TGF-βr2-/-mice were given Smad signaling agonist SRI-011381(SRI)intramus-cular injection.All mice were mainly divided into the following groups:control group,SM TGF-βr2-/-group and SM TGF-βr2-/-+SRI group.Twenty-four mice were selected in each group.RT-qPCR and immunofluorescence staining were used to detect the relative mRNA level,protein expression of inflammatory cytokines and low-density lipoprotein receptor-related protein 1(LRP1),respectively,while the relative protein expression of myosin heavy chain 3(MHY3)and embryonic myosine heavy chain(eMHC)in damaged muscle was detected by Western blot and immunofluorescence staining.In vi-tro,after being extracted from neonatal mice,myogenic precursor cells(MPCs)were cultured in an pro-inflammatory mi-lieu and treated with SRI,recombinant mouse extracellular matrix protein 1(rmECM1)alone or in combination.Hereby,they were divided into the following seven groups:control-MPCs group,control-MPCs+LPS group,TGF-βr2-/--MPCs group,TGF-βr2-/--MPCs+LPS group,TGF-βr2-/--MPCs+LPS+SRI group,TGF-βr2-/--MPCs+LPS+rmECM1 group,and TGF-βr2-/--MPCs+LPS+SRI+rmECM1 group.Six mice were selected in each group.RT-qPCR and Western blot were used to detect the relative mRNA level,protein expression of major histocompatibility complex class I molecules(MHC-I/H-2Kb),major histocompatibility complex class II molecules(MHC-II/H2-Eα),Toll-like receptor 3(TLR3),and LRP1.And the relative protein expression of MoyD and myogenin in myotubes was detected by immunofluorescence staining.RE-SULTS:In vivo,compared with control group,SM TGF-βr2-/-group showed the significant upregulation of pro-inflamma-tory cytokines(P<0.05),and the opposite trend of anti-inflammatory cytokines,LRP1,MHY3,eMHC in the injured muscle(P<0.05),with delayed regeneration and repair of myofibers.In vitro,compared with control-MPCs+LPS group,LRP1,MoyD and myogenin significantly downregulated in TGF-βr2-/--MPCs+LPS group,but the downregulation trend was corrected after giving SRI treatment(P<0.05).In addition,compared with the TGF-βr2-/--MPCs+LPS group,the combi-nation of rmECM1 and SRI significantly upregulated the protein expression of MyoD and myogenin(P<0.05).CONCLU-SION:In a mouse model of acute skeletal myositis,intrinsic TGF-β signaling specifically in myofibers regulates local im-mune behavior.It promotes the expression of LRP1 in damaged muscle via Smad2/3 signaling,and LRP1 can then fully bind to ECM1,thereby facilitating muscle regeneration and repair,and improving the prognosis of acute skeletal myositis.

Purpose To explore the application value of magnetic resonance feature tracking to assess early heart failure in different rat models.Materials and Methods Fifty male SD rats were randomly divided into four groups:the control group(8 rats),the isoproterenol high dose(ISO-H)group(14 rats),the isoproterenol low dose(ISO-L)group(14 rats)and the coronary artery ligation surgery(myocardial infarction,MI)group(14 rats).The ISO-H group received intraperitoneal injections of ISO at 100 mg/kg once a week for two consecutive weeks.The ISO-L group received continuous intraperitoneal injections of ISO at 5 mg/(kg·d)for 14 days.The MI group underwent coronary ligation to establish a heart failure rat model.Scans using a 7T cardiac MRI cine sequence were performed after two weeks and four weeks.Heart function parameters and myocardial strain parameters were obtained using CVI 42 software.Serum markers of myocardial injury,including creatine kinase,creatine kinase-MB,lactate dehydrogenase and α-hydroxybutyric dehydrogenase levels,were measured.HE staining and Masson staining were used to detect myocardial pathological changes.Results Compared with the control group,the ISO-H group,ISO-L group and MI group showed significant changes in left ventricular ejection fraction(t=-2.387,P=0.04;t=5.622,P<0.001;t=17.715,P<0.001)and overall radial strain(t=-4.570,P=0.001;t=6.587,P<0.001;t=10.649,P<0.001)after two weeks.Compared with the control group,the ISO-H group,ISO-L group and MI group showed varying degrees of reduction in left ventricular ejection fraction(t=4.834,P=0.001;t=10.325,P<0.001;t=26.286,P<0.001)and overall radial strain(t=3.890,P=0.003;t=9.547,P<0.001;t=13.751,P<0.001)after four weeks.The serum markers of myocardial injury,including creatine kinase,creatine kinase-MB,lactate dehydrogenase and α-hydroxybutyric dehydrogenase levels,were significantly elevated(t=-93.373--11.019,P<0.001),and pathological staining revealed that all model groups exhibited varying degrees of myocardial cell hypertrophy,necrosis and interstitial fibrosis compared with the control group.Conclusion Magnetic resonance feature tracking has guiding significance for the dynamic monitoring,early diagnosis and prognosis assessment of heart failure in different rat models.

AIM:To investigate the effect of intrinsic specific transforming growth factor-β(TGF-β)signaling on regeneration and repair of myofibers in acute skeletal myositismice model induced by cardiotoxin(CTX).METHODS:One hundred and eighty-six wild C57BL/6 mice and one hundred and thirty-eight mice with conditional knockout of TGF-β receptor 2(TGF-βr2)in myofibers(SM TGF-βr2-/-mice)were selected.CTX injection to anterior tibial muscle(TA)in-duced acute myoinjury in mice.Some SM TGF-βr2-/-mice were given Smad signaling agonist SRI-011381(SRI)intramus-cular injection.All mice were mainly divided into the following groups:control group,SM TGF-βr2-/-group and SM TGF-βr2-/-+SRI group.Twenty-four mice were selected in each group.RT-qPCR and immunofluorescence staining were used to detect the relative mRNA level,protein expression of inflammatory cytokines and low-density lipoprotein receptor-related protein 1(LRP1),respectively,while the relative protein expression of myosin heavy chain 3(MHY3)and embryonic myosine heavy chain(eMHC)in damaged muscle was detected by Western blot and immunofluorescence staining.In vi-tro,after being extracted from neonatal mice,myogenic precursor cells(MPCs)were cultured in an pro-inflammatory mi-lieu and treated with SRI,recombinant mouse extracellular matrix protein 1(rmECM1)alone or in combination.Hereby,they were divided into the following seven groups:control-MPCs group,control-MPCs+LPS group,TGF-βr2-/--MPCs group,TGF-βr2-/--MPCs+LPS group,TGF-βr2-/--MPCs+LPS+SRI group,TGF-βr2-/--MPCs+LPS+rmECM1 group,and TGF-βr2-/--MPCs+LPS+SRI+rmECM1 group.Six mice were selected in each group.RT-qPCR and Western blot were used to detect the relative mRNA level,protein expression of major histocompatibility complex class I molecules(MHC-I/H-2Kb),major histocompatibility complex class II molecules(MHC-II/H2-Eα),Toll-like receptor 3(TLR3),and LRP1.And the relative protein expression of MoyD and myogenin in myotubes was detected by immunofluorescence staining.RE-SULTS:In vivo,compared with control group,SM TGF-βr2-/-group showed the significant upregulation of pro-inflamma-tory cytokines(P<0.05),and the opposite trend of anti-inflammatory cytokines,LRP1,MHY3,eMHC in the injured muscle(P<0.05),with delayed regeneration and repair of myofibers.In vitro,compared with control-MPCs+LPS group,LRP1,MoyD and myogenin significantly downregulated in TGF-βr2-/--MPCs+LPS group,but the downregulation trend was corrected after giving SRI treatment(P<0.05).In addition,compared with the TGF-βr2-/--MPCs+LPS group,the combi-nation of rmECM1 and SRI significantly upregulated the protein expression of MyoD and myogenin(P<0.05).CONCLU-SION:In a mouse model of acute skeletal myositis,intrinsic TGF-β signaling specifically in myofibers regulates local im-mune behavior.It promotes the expression of LRP1 in damaged muscle via Smad2/3 signaling,and LRP1 can then fully bind to ECM1,thereby facilitating muscle regeneration and repair,and improving the prognosis of acute skeletal myositis.

A throat swab sample from a pediatric case in Tongzhou District, Beijing was identified as enterovirus; the patient was a 1-year-and-8-month-old male sporadic case. Whole-genome sequencing revealed a viral genome length of 7 436 bp. BLAST alignment confirmed the serotype as EV-D68. Phylogenetic analysis of the whole genome indicated that this strain belongs to the B3 clade, showing closer genetic proximity to the 2018 Shanghai strain MW697453 with 99.53% whole-genome nucleotide homology. Genetic and amino acid variation analysis demonstrated that the B3 subclade to which this strain belongs exhibits a nucleotide deletion at positions 718–726, differing from deletion sites observed in other B3 clade strains. A key neuropathogenic amino acid site, T650A, was found to have undergone mutation. Recombination analysis confirmed no cross-clade recombination events in this strain. This study conducted genetic characterization of the strain's evolutionary relationship with EV-D68 strains from different regions and years in China, providing data support for formulating prevention and control measures against EV-D68 infection.

Purpose To explore the application value of magnetic resonance feature tracking to assess early heart failure in different rat models.Materials and Methods Fifty male SD rats were randomly divided into four groups:the control group(8 rats),the isoproterenol high dose(ISO-H)group(14 rats),the isoproterenol low dose(ISO-L)group(14 rats)and the coronary artery ligation surgery(myocardial infarction,MI)group(14 rats).The ISO-H group received intraperitoneal injections of ISO at 100 mg/kg once a week for two consecutive weeks.The ISO-L group received continuous intraperitoneal injections of ISO at 5 mg/(kg·d)for 14 days.The MI group underwent coronary ligation to establish a heart failure rat model.Scans using a 7T cardiac MRI cine sequence were performed after two weeks and four weeks.Heart function parameters and myocardial strain parameters were obtained using CVI 42 software.Serum markers of myocardial injury,including creatine kinase,creatine kinase-MB,lactate dehydrogenase and α-hydroxybutyric dehydrogenase levels,were measured.HE staining and Masson staining were used to detect myocardial pathological changes.Results Compared with the control group,the ISO-H group,ISO-L group and MI group showed significant changes in left ventricular ejection fraction(t=-2.387,P=0.04;t=5.622,P<0.001;t=17.715,P<0.001)and overall radial strain(t=-4.570,P=0.001;t=6.587,P<0.001;t=10.649,P<0.001)after two weeks.Compared with the control group,the ISO-H group,ISO-L group and MI group showed varying degrees of reduction in left ventricular ejection fraction(t=4.834,P=0.001;t=10.325,P<0.001;t=26.286,P<0.001)and overall radial strain(t=3.890,P=0.003;t=9.547,P<0.001;t=13.751,P<0.001)after four weeks.The serum markers of myocardial injury,including creatine kinase,creatine kinase-MB,lactate dehydrogenase and α-hydroxybutyric dehydrogenase levels,were significantly elevated(t=-93.373--11.019,P<0.001),and pathological staining revealed that all model groups exhibited varying degrees of myocardial cell hypertrophy,necrosis and interstitial fibrosis compared with the control group.Conclusion Magnetic resonance feature tracking has guiding significance for the dynamic monitoring,early diagnosis and prognosis assessment of heart failure in different rat models.

Objective To investigate the relationship between interlukin-10(IL-10)gene polymorphisms and acute kidney injury(AKI)and its related mechanisms.Methods A prospective study was conducted on 100 patients with AKI admitted to our hospital from March 2021 to March 2023,who were selected as the study group.Following the 1∶1 pairing principle,100 healthy individuals who underwent physical examinations during the same period were included as the control group.General data,IL-10 genotypes,and gene frequency distributions of the two groups were compared.The factors influencing AKI were assessed using a logistic regression analysis,and the interaction between IL-10gene polymorphisms and conventional risk factors was analyzed using multifactor dimension reduction(MDR).Results The quantitative values of serum creatinine(sCr),blood urea nitrogen(BUN),and urine protein in the study group were higher than those in the control group.The proportion of the GG genotype at IL-10-1082 was lower than that in the control group,and the proportions of the AA genotype and A allele were higher than those in the control group(P<0.05).Logistic regression analysis showed that BUN(OR=4.487),urinary protein quantitation(OR=5.905),sCr(OR=3.573),AA genotype at position 1082(OR=4.823),and the A allele(OR=4.479)were risk factors for AKI(P<0.05).Interaction display,IL-10-1082 polymorphism×sCr,IL-10-1082 polymor-phism×BUN,IL-10-1082 polymorphism×urinary protein quantification,and IL-10-1082 polymorphism×sCr×BUN×urinary protein quantification models exhibited good accuracy and cross-consistency(P<0.05).Conclusion IL-10gene polymorphisms are strongly associated with the development of AKI.

Purpose To explore the cardioprotective effect of cang-ai volatile oil(CAVO)on rats with cardiac function impairment model under low-pressure and low-oxygen environment in Tibet Plateau based on 7.0T cardiovascular magnetic resonance(CMR)imaging.Materials and Methods Forty SD rats were randomly divided into the normal group,the high altitude model group,the CAVO-treated group and the rhodiola rosea-treated group,with 10 rats in each group.Except for the normal group,the rats in other groups were transferred from the plain(500 m above sea level)to the Tibet Plateau(4 250 m above sea level)for two months,and then administered with the corresponding drugs by gavage for 14 d.The left ventricle function was measured by using a 7.0T high-field strength CMR and myocardial strain was analysed by using tissue tracing technique.HE staining was used to observe the morphology of cardiomyocytes,Masson staining to observe interstitial fibrosis,wheat germ agglutinin staining to observe cardiomyocyte hypertrophy,and transmission electron microscopy to observe the morphological changes of mitochondria in each group.Serum levels of creatine kinase,creatine kinase isoenzyme,lactate dehydrogenase,cardiac troponin T,superoxide dismutase,malondialdehyde and glutathione peroxidase were detected.Intracellular reactive oxygen species levels were detected using flow cytometry.Results The left ventricular ejection fraction of rats in the CAVO-treated group was higher than that of the high altitude model group[(66.61±1.38)％vs.(60.94±3.21)％;t=3.969,P=0.032];meanwhile,the global circumferential strain of the left ventricle in the CAVO-treated group was higher than that of the high altitude model group(-25.68±1.30 vs.-22.84±1.17;t=3.967,P=0.003).HE,Masson and wheat germ agglutinin staining showed hypertrophy and necrosis as well as interstitial fibrosis and ultrastructural disruption of cardiomyocytes in the high altitude model group,which improved after CAVO treatment.The level of cardiac troponin T in the serum of rats with CAVO treatment group was significantly decreased compared with that of the high altitude model group[(314.03±20.05)pg/ml vs.(518.30±18.13)pg/ml;1=13.090,P=0.001].Conclusion CAVO treatment can reduce cardiac injury caused by low-pressure hypoxia in high altitude,and its effect can be detected dynamically and non-invasively by 7.0T high-field strength CMR.

Objective To investigate the effect of E2 signaling in myofibers on muscular macrophage efferocytosis in mice with cardiotoxin-induced acute skeletal muscle injury.Methods Female wild-type C57BL/6 mice with and without ovariectomy and male C57BL/6 mice were given a CTX injection into the anterior tibial muscle to induce acute muscle injury,followed by intramuscular injection of β-estradiol(E2)or 4-hydroxytamoxifen(4-OHT).The changes in serum E2 of the mice were detected using ELISA,and the number,phenotypes,and efferocytosis of the macrophages in the inflammatory exudates and myofiber regeneration and repair were evaluated using immunofluorescence staining and flow cytometry.C2C12 cells were induced to differentiate into mature myotubes,which were treated with IFN-γ for 24 before treatment with β-Estradiol or 4-OHT.The treated myotubes were co-cultured with mouse peritoneal macrophages in a 1:2 ratio,followed by addition of PKH67-labeled apoptotic mouse mononuclear spleen cells induced by UV irradiation,and macrophage efferocytosis was observed using immunofluorescence staining and flow cytometry.Results Compared with the control mice,the female mice with ovariectomy showed significantly increased mononuclear macrophages in the inflammatory exudates,with increased M1 cell percentage,reduced M2 cell percentage and macrophage efferocytosis in the injured muscle,and obviously delayed myofiber regeneration and repair.In the cell co-culture systems,treatment of the myotubes with β-estradiol significantly increased the number and proportion of M2 macrophages and macrophage efferocytosis,while 4-OHT treatment resulted in the opposite changes.Conclusion In injured mouse skeletal muscles,myofiber E2 signaling promotes M1 to M2 transition to increase macrophage efferocytosis,thereby relieving inflammation and promoting muscle regeneration and repair.