OBJECTIVE: To assess the efficacy of Qingda Granule (QDG) in ameliorating hypertension-induced cardiac damage and investigate the underlying mechanisms involved. METHODS: Twenty spontaneously hypertensive rats (SHRs) were used to develope a hypertension-induced cardiac damage model. Another 10 Wistar Kyoto (WKY) rats were used as normotension group. Rats were administrated intragastrically QDG [0.9 g/(kg•d)] or an equivalent volume of pure water for 8 weeks. Blood pressure, histopathological changes, cardiac function, levels of oxidative stress and inflammatory response markers were measured. Furthermore, to gain insights into the potential mechanisms underlying the protective effects of QDG against hypertension-induced cardiac injury, a network pharmacology study was conducted. Predicted results were validated by Western blot, radioimmunoassay immunohistochemistry and quantitative polymerase chain reaction, respectively. RESULTS: The administration of QDG resulted in a significant decrease in blood pressure levels in SHRs (P<0.01). Histological examinations, including hematoxylin-eosin staining and Masson trichrome staining revealed that QDG effectively attenuated hypertension-induced cardiac damage. Furthermore, echocardiography demonstrated that QDG improved hypertension-associated cardiac dysfunction. Enzyme-linked immunosorbent assay and colorimetric method indicated that QDG significantly reduced oxidative stress and inflammatory response levels in both myocardial tissue and serum (P<0.01). CONCLUSIONS Both network pharmacology and experimental investigations confirmed that QDG exerted its beneficial effects in decreasing hypertension-induced cardiac damage by regulating the angiotensin converting enzyme (ACE)/angiotensin II (Ang II)/Ang II receptor type 1 axis and ACE/Ang II/Ang II receptor type 2 axis.
Animals ; Drugs, Chinese Herbal/therapeutic use* ; Hypertension/pathology* ; Renin-Angiotensin System/drug effects* ; Rats, Inbred SHR ; Oxidative Stress/drug effects* ; Male ; Rats, Inbred WKY ; Blood Pressure/drug effects* ; Myocardium/pathology* ; Rats ; Inflammation/pathology*

OBJECTIVE: To explore the functional roles and underlying mechanisms of neferine in the context of angiotensin II (Ang II)-induced hypertension and vascular dysfunction. METHODS: Male mice were infused with Ang II to induce hypertension and randomly divided into treatment groups receiving neferine or a control vehicle based on baseline blood pressure using a random number table method. The hypertensive mouse model was constructed by infusing Ang II via a micro-osmotic pump (500 ng/kg per minute), and neferine (0.1, 1, or 10 mg/kg), valsartan (10 mg/kg), or double distilled water was administered intragastrically once daily for 6 weeks. A non-invasive blood pressure system, ultrasound, and hematoxylin and eosin staining were performed to assess blood pressure and vascular changes. RNA sequencing and network pharmacology were employed to identify differentially expressed transcripts (DETs) and pathways. Vascular ring tension assay was used to test vascular function. A7R5 cells were incubated with neferine for 24 h and then treated with Ang II to record the real-time Ca²⁺ concentration by confocal microscope. Immunohistochemistry (IHC) and Western blot were used to evaluate vasorelaxation, calcium, and the extracellular signal-regulated kinase (ERK)1/2 pathway. RESULTS: Neferine treatment effectively mitigated the elevation in blood pressure, pulse wave velocity, aortic thickening in the abdominal aorta of Ang II-infused mice (P<0.05). RNA sequencing and network pharmacology analysis identified 355 DETs that were significantly reversed by neferine treatment, along with 25 potential target genes, which were further enriched in multiple pathways and biological processes, such as ERK1 and ERK2 cascade regulation, calcium pathway, and vascular smooth muscle contraction. Further investigation revealed that neferine treatment enhanced vasorelaxation and reduced Ca²⁺-dependent contraction of abdominal aortic rings, independent of endothelium function (P<0.05). The underlying mechanisms were mediated, at least in part, via suppression of receptor-operated channels, store-operated channels, or voltage-operated calcium channels. Neferine pre-treatment demonstrated a reduction in intracellular Ca²⁺ release in Ang II stimulated A7R5 cells. IHC staining and Western blot confirmed that neferine treatment effectively attenuated the upregulation of p-ERK1/2 both in vivo and in vitro, which was similar with treatment of ERK1/2 inhibitor PD98059 (P<0.05). CONCLUSIONS Neferine remarkably alleviates Ang II-induced elevation of blood pressure, vascular dysfunction, and pathological changes in the abdominal aorta. This beneficial effect is mediated by the modulation of multiple pathways, including calcium and ERK1/2 pathways.
Animals ; Angiotensin II ; Male ; Benzylisoquinolines/therapeutic use* ; Network Pharmacology ; Blood Pressure/drug effects* ; Sequence Analysis, RNA ; Mice ; Hypertension/chemically induced* ; Mice, Inbred C57BL ; Calcium/metabolism*

OBJECTIVE: To identify the underlying mechanism by which quercetin (Que) alleviates sepsis-related acute respiratory distress syndrome (ARDS). METHODS: In vivo, C57BL/6 mice were assigned to sham, cecal ligation and puncture (CLP), and CLP+Que (50 mg/kg) groups (n=15 per group) by using a random number table. The sepsisrelated ARDS mouse model was established using the CLP method. In vitro, the murine alveolar macrophages (MH-S) cells were classified into control, lipopolysaccharide (LPS), LPS+Que (10 μmol/L), and LPS+Que+acetylcysteine (NAC, 5 mmol/L) groups. The effect of Que on oxidative stress, inflammation, and apoptosis in mice lungs and MH-S cells was determined, and the mechanism with reactive oxygen species (ROS)/p38 mitogen-activated protein kinase (MAPK) pathway was also explored both in vivo and in vitro. RESULTS: Que alleviated lung injury in mice, as reflected by a reversal of pulmonary histopathologic changes as well as a reduction in lung wet/dry weight ratio and neutrophil infiltration (P<0.05 or P<0.01). Additionally, Que improved the survival rate and relieved gas exchange impairment in mice (P<0.01). Que treatment also remarkedly reduced malondialdehyde formation, superoxide dismutase and catalase depletion, and cell apoptosis both in vivo and in vitro (P<0.05 or P<0.01). Moreover, Que treatment diminished the release of inflammatory factors interleukin (IL)-1β, tumor necrosis factor-α, and IL-6 both in vivo and in vitro (P<0.05 or P<0.01). Mechanistic investigation clarifified that Que administration led to a decline in the phosphorylation of p38 MAPK in addition to the suppression of ROS expression (P<0.01). Furthermore, in LPS-induced MH-S cells, ROS inhibitor NAC further inhibited ROS/p38 MAPK pathway, as well as oxidative stress, inflammation, and cell apoptosis on the basis of Que treatment (P<0.05 or P<0.01). CONCLUSION Que was found to exert anti-oxidative, anti-inflammatory, and anti-apoptotic effects by suppressing the ROS/p38 MAPK pathway, thereby conferring protection for mice against sepsis-related ARDS.
Animals ; Sepsis/drug therapy* ; Quercetin/therapeutic use* ; Respiratory Distress Syndrome/enzymology* ; p38 Mitogen-Activated Protein Kinases/metabolism* ; Mice, Inbred C57BL ; Reactive Oxygen Species/metabolism* ; Apoptosis/drug effects* ; Male ; Oxidative Stress/drug effects* ; MAP Kinase Signaling System/drug effects* ; Lung/drug effects* ; Mice ; Lipopolysaccharides ; Macrophages, Alveolar/pathology* ; Inflammation/pathology* ; Protective Agents/therapeutic use*

Ischemic stroke (IS) is a globally life-threatening disease. Presently, few therapeutic medicines are available for treating IS, and rt-PA is the only drug approved by the US Food and Drug Administration (FDA) in the US. In fact, many agents showing excellent neuroprotection but no blood flow-improving activity in animals have not achieved ideal clinical efficacy, while thrombolytic drugs only improving blood flow without neuroprotection have limited their wider application. To address these challenges and meet the huge unmet clinical need, we have designed and identified a novel compound AAPB with dual effects of neuroprotection and cerebral blood flow improvement. AAPB significantly reduced cerebral infarction and neural function deficit in tMCAO rats, pMCAO rats, and IS rhesus monkeys, as well as displayed exceptional safety profiles and excellent pharmacokinetic properties in rats and dogs. AAPB has now entered phase I of clinical trials fighting IS in China.

Objective To investigate the effectiveness and safety of biological patch in minimally invasive inguinal hernia surgery.Methods From July 2019 to July 2021,100 inguinal hernia patients were divided into two groups based on the actual type of patch received.Biological patch was used in the experimental group(50 cases),and polypropylene patch was used in the control group(50 cases).The operation method was laparoscopic transabdominal preperitoneal(TAPP)hernia repair.Operating time,bleeding volume during operation,hospital stay after operation,hernia recurrence rate,incision infection rate,patch infection rate,and complication rate were compared between the two groups.Results Two groups of patients had no recurrence of hernia within 3 years after surgery,and didn't experience incision infection or patch infection after surgery.There were no significant differences in operating time,bleeding volume during operation,hospital stay after operation,seroma rate and chronic pain rate between the two groups(P>0.05).The incidence rate of foreign body sensation in the control group was higher than that in the experimental group,with statistical difference(P<0.05).Conclusion In conclusion,biological patch is safe and effective in laparoscopic TAPP hernia repair,and can reduce the incidence of postoperative foreign body sensation,providing a new option for laparoscopic inguinal hernia repair.

This study was aimed at understanding the prevalence and drug resistance status of Salmonella of waterfowl ori-gin in the Guangdong region in the past decade,to guide prevention and control efforts.The drug-sensitive paper slide method was used to conduct drug susceptibility testing on 314 waterfowl-originating Salmonella strains isolated from 238 waterfowl farms in the Guangdong region from 2013 to 2023.The isolated Salmonella strains were most resistant to penicillin,amoxicil-lin,cefradine,and cefazolin in the β-lactam group;sulphadoxine dimethylpyrimidine in the sulphonamide group;and tetracy-cline in the tetracycline group.The resistance rates ranged from 73.57％to 89.49％.The highest sensitivity was observed to amikacin,gentamicin,and kanamycin in the aminoglycoside group,and norfloxacin in the quinolone group,with susceptibility rates all exceeding 50％.The 280 strains of Salmonella showed multi-drug resistance to six classes of antimicrobial drugs and high resistance(as much as 60.83％)to five drug classes.Correlation analysis revealed the highest correlations for florfenicol with gentamicin,and for amoxicillin with penicillin(r=0.650 for both),followed by gentamicin with kanamycin(r=0.620).Salmonella resistance in waterfowl in Guangdong Province was generally severe and showed a complex pattern of drug resist-ance.Detection of waterfowl pathogens should be strengthened to prevent the spread of drug-resistant bacteria and support ra-tional use of antibiotics.This work provides a reference for Salmonella prevention and control in waterfowl farms.

Objective The aim of this study is to analyze the expression level of cell-free DNA(cf-DNA),a biomarker of neutrophil extracellular traps(NETs),in children with Mycoplasma pneumoniae pneumonia(MPP),and to explore the predictive efficacy of cf-DNA(as a marker of NETs)for the severity of MPP in these children.Methods A total of 115 children with MPP were prospectively selected as the MPP group.Based on the disease severity,the MPP group was categorized into the mild group(n=75)and the severe group(n=40).During the same period,50 healthy children undergoing physical examinations were selected as the control group.The levels of serum cf-DNA in the MPP group and the control group,as well as the levels of C-reactive protein(CRP),D-dimer,lactate dehydrogenase(LDH),interleukin-6(IL-6),interferon-γ(IFN-γ),and tumor necrosis factor-α(TNF-α)in the MPP group were detected.The differences in the levels of serum cf-DNA and related inflammatory factors among the groups were compared,and the role of serum cf-DNA in evaluating the severity of MPP was analyzed.Results The level of serum cf-DNA in children of the MPP group was notably higher than that in the control group,with a more significant elevation observed in the severe group(P<0.05).The levels of CRP,D-dimer,LDH,IL-6,IFN-γ,and TNF-α were all higher in the severe group than in the mild group(P<0.05).Multivariate logistic regression analysis showed that the increased levels of serum cf-DNA,CRP,and IL-6 were closely related to the severity of MPP(P<0.05).The results of receiver operating characteristic(ROC)curve analysis showed that the area under the curve(AUC)of the combination of serum cf-DNA,CRP,and IL-6 for predicting severe MPP was 0.981,which was higher than that of each index alone(P<0.05).Conclusions Serum cf-DNA(as a marker of NETs)is closely related to the severity of MPP in children.The combined detection of cf-DNA,CRP,and IL-6 is more beneficial for assessing the severity of MPP in children.

Aim To explore the effects of high glucose on KIAA0753 and CCSAP and the relationship between KIAA0753 and CCSAP and osteogenic differentiation and calcium signaling pathway under high glucose con-ditions.Methods Mouse embryonic osteoblast pre-cursor cells(MC3T3-E1)were induced by osteoblast medium with glucose concentrations of 5.5 and 25 mmol·L-1,and the protein expressions of KIAA0753 and CCSAP were detected by Western blot.The over-expressed plasmid was transfected into human embry-onic kidney cells(HEK-293T),and the interaction between KIAA0753 and CCSAP was detected by co-im-munoprecipitation.MC3T3-E1 cells were treated in the osteogenic medium with different glucose concentrations and induction times.Alkaline phosphatase(ALP)ac-tivity was detected with a kit.The expression of KI-AA0753,CCSAP,osteopontin,osteocalcin,and other proteins were assessed using Western blot.and then 5.5,25 mmol·L-1,and 25 mmol·L-1+OE-CCSAP three groups of MC3T3-E1 cells were set.The expression of KIAA0753,OCN,OPN protein,and ALP activity were detected successively.The diabetic mouse model dataset in Gene Expression Omnibus was used to screen differential genes for bioinformatics a-nalysis.MC3T3-E1 cells were set up in three groups,5.5,25 mmol·L-1,and 25 mmol·L-1+OE-KI-AA0753,respectively.The calcium/calmodulin-de-pendent protein kinase Ⅱ beta(CAMK2B)and phos-pholamban(PLN)were detected by Western blot.Re-sults Compared with the 5.5 mmol·L-1 group,25 mmol·L-1 inhibited the expression of KIAA0753 and CCSAP proteins in osteoblasts,and there was an inter-action between KIAA0753 and CCSAP.At the same time,25 mmol·L-1 also inhibited the expression of ALP,OPN,and OCN proteins in osteoblasts.Overex-pression of CCSAP at 25 mmol·L-1 up-regulated the expression of KIAA0753,OCN,OPN,and ALP.The differential genes of the diabetic mouse model were mainly concentrated in the aspects of"signal receptor and signal regulation".25 mmol·L-1 glucose inhibi-ted the expression of CAMK2B and PLN proteins in os-teoblasts,and overexpression of KIAA0753 at 25 mmol·L-1 upregulated the expression of CAMK2B and PLN proteins.Conclusions High glucose inhibits the expression of KIAA0753 and CCSAP protein,inhibits the osteogenic differentiation and calcium signaling in mouse embryonic osteoblast precursor cells,overex-pression of CCSAP saves the inhibitory effect of high glucose on osteogenic differentiation,and overexpres-sion of KIAA0753 reverses the inhibitory effect of high glucose on calcium signaling pathway.

Objective To investigate the bidirectional causal relationship between circulating amino acid levels and the risk of myasthenia gravis(MG)using Mendelian randomization(MR).Methods A two-sample Mendelian randomization a-nalysis was conducted using publicly available genome-wide association study(GWAS)genetic data,with validation from GWAS data from different sources to assess the robustness of the results.Five models were used for the two-sample bidirectional MR anal-ysis,and odds ratios(OR)were calculated to evaluate the causal relationship between the levels of nine circulating amino acids and MG risk.Sensitivity analyses,heterogeneity tests,and pleiotropy tests were performed to assess the robustness of the results.The causal effect estimated by the inverse variance weighted(IVW)method was the primary result,and the IVW-estimated causal effects were further validated using data from different GWAS sources to assess robustness.Results Genetically predicted high-er circulating glutamine levels were significantly associated with a lower risk of MG[OR(95％CI)=0.696(0.524,0.926),P=0.012 7,IVW model].Validation analyses using GWAS data from various sources also demonstrated a significant negative associa-tion between genetically predicted higher circulating glutamine levels and MG risk[OR(95％CI)=0.321(0.178,0.581),P=1.67x10-1,IVW model].Moreover,genetically predicted higher MG risk was associated with lower levels of circulating glutamine and alanine(β=-0.178±0.009,P=0.049;β=-0.013±0.007,P=0.048,IVW model,respectively).Conclusion Genetic evidence reveals a potential bidirectional causal relationship between circulating amino acid levels and MG risk.Further studies are required to elucidate the mechanisms underlying this relationship.

Objective To explore the diagnostic value of electrocardiogram parameters combined with serum microRNA(miR)-322 and miR-568 levels in patients with chronic heart failure(CHF)complicated with ventricular arrhythmias(VA).Methods A total of 230 CHF patients admitted to Zhangjiakou First Hospital from April 2020 to April 2023 were selected as the study objects,including 120 VA patients(VA group)and 110 non VA patients(non VA group),and 110 patients who underwent health examinations in Zhangjiakou First Hospital during the same period as the control group.Compared three sets of general information,electrocardiogram parameters,left ventricular ejection fraction(LVEF),cardiac function grade and serum miR-322 and miR-568 levels.Logistic regression analyzed the influencing factors of VA in CHF patients.Receiver operating characteristic curve(ROC)analyzed electrocardiogram parameters combined with serum miR-322 and miR-568 for the diagnostic value of VA in CHF patients.Results The three groups showed statistically obvious differences in classification of nyha heart function(NYHA)and left ventricular ejection fraction(LVEF)(F=6.033,691.853,all P<0.05).The electrocardiogram parameters of CHF patients,including QT interval dispersion(QTD),QRS wave duration and corrected QT systolic time(QTc)were obviously higher than those in the control group(t=16.539,17.709,14.414),and the VA group were obviously higher than the non VA group(q=10.984,7.794,10.174),and the differences were statistically significan(all P<0.05).The serum levels of miR-322 and miR-568 were obviously lower than those in the control group(t=23.719,17.359).and the VA group were obviously lower than the non VA group(q=10.345,9.941),the differences were statistically significant(all P<0.05).Logistic regression analysis showed that NYHA grading,LVEF,QTD,QRS,wave duration,QTc,serum miR-322 and miR-568 levels were all influencing factors in the complication of VA in patients with CHF(Wald χ2=4.267～9.839,all P<0.05).The combination of electrocardiogram parameters and serum miR-322,miR-568 was better diagnosing CHF patients with concurrent VA that QTD,QRS duration,QTc and serum miR-332,miR-568 were measured separately.Conclusion Serum levels of miR-322 and miR-568 are obviously reduced,and the combination of electrocardiogram parameters with serum miR-322 and miR-568 has high diagnostic value for CHF patients with VA.