ObjectiveTo construct an animal model of respiratory syncytial virus（RSV）-infected pneumonia suitable for preclinical studies. MethodsThe virulence of RSV to the four cell lines was observed by cytopathic effect （CPE）， and 50% tissue culture infective dose（TCID₅₀） was calculated. Twenty BALB/c mice were randomly divided into a normal group and a model group. Six BALB/c-hIGF1R mice served as the humanized IGF1R model group. Except for the normal group， the other groups received intranasal RSV infection on days 1 and 3 to establish a viral pneumonia model. The efficacy of establishing an RSV-induced pneumonia animal model based on humanized insulin-like growth factor 1 receptor （IGF1R） mice was evaluated by measuring organ indices， peripheral blood lymphocyte percentages， pulmonary pathology and imaging， and pulmonary viral load. Additionally， ten BALB/c mice served as normal group， and thirty-two BALB/c-hIGF1R mice were randomly assigned to humanized IGF1R model group， ribavirin group （82.5 mg·kg^-¹·d^-¹）， and high and low dose groups of Lianhua Qingwen （3.3 mg·kg^-¹·d^-¹ ， 1.65 mg·kg^-¹·d^-¹）， with 8 mice per group. The viral load in lung tissue was measured after ribavirin and Lianhua Qingwen intervention， and the model was applied to the evaluation of anti-RSV drugs. ResultsIn the lungs of the humanized IGF1R model group， large solid and diffuse ground-glass shadows were seen， and the lung volume was significantly increased （P<0.01）. The lung index was significantly increased （P<0.01）， and both the spleen index and thymus index were significantly decreased （P<0.01）. The percentages of CD3⁺ and CD4⁺T cells were significantly decreased （P<0.05）， and there was a large amount of inflammation and stasis in the perivascular area of the lung tissue， which was predominantly characterized by lymphocytes. The endothelium of blood vessels was partially detached， with a small number of eosinophils. After infecting BALB/c-hIGF1R mice with RSV， the expression of viral nucleic acids in the lung tissue of the mice was significantly increased， with significant differences compared with the normal group （P<0.01）. The expression of viral nucleic acids in the ribavirin group and the high and low dose groups of Lianhua Qingwen was significantly reduced， with significant differences compared with the normal group （P<0.01）. ConclusionHumanized IGF1R mice are more susceptible to respiratory SVC， and the animal model of RSV-infected pneumonia based on humanized IGF1R mice was successfully constructed， which is suitable for the evaluation of anti-RSV drugs.

ObjectiveTo construct an animal model of respiratory syncytial virus（RSV）-infected pneumonia suitable for preclinical studies. MethodsThe virulence of RSV to the four cell lines was observed by cytopathic effect （CPE）， and 50% tissue culture infective dose（TCID₅₀） was calculated. Twenty BALB/c mice were randomly divided into a normal group and a model group. Six BALB/c-hIGF1R mice served as the humanized IGF1R model group. Except for the normal group， the other groups received intranasal RSV infection on days 1 and 3 to establish a viral pneumonia model. The efficacy of establishing an RSV-induced pneumonia animal model based on humanized insulin-like growth factor 1 receptor （IGF1R） mice was evaluated by measuring organ indices， peripheral blood lymphocyte percentages， pulmonary pathology and imaging， and pulmonary viral load. Additionally， ten BALB/c mice served as normal group， and thirty-two BALB/c-hIGF1R mice were randomly assigned to humanized IGF1R model group， ribavirin group （82.5 mg·kg^-¹·d^-¹）， and high and low dose groups of Lianhua Qingwen （3.3 mg·kg^-¹·d^-¹ ， 1.65 mg·kg^-¹·d^-¹）， with 8 mice per group. The viral load in lung tissue was measured after ribavirin and Lianhua Qingwen intervention， and the model was applied to the evaluation of anti-RSV drugs. ResultsIn the lungs of the humanized IGF1R model group， large solid and diffuse ground-glass shadows were seen， and the lung volume was significantly increased （P<0.01）. The lung index was significantly increased （P<0.01）， and both the spleen index and thymus index were significantly decreased （P<0.01）. The percentages of CD3⁺ and CD4⁺T cells were significantly decreased （P<0.05）， and there was a large amount of inflammation and stasis in the perivascular area of the lung tissue， which was predominantly characterized by lymphocytes. The endothelium of blood vessels was partially detached， with a small number of eosinophils. After infecting BALB/c-hIGF1R mice with RSV， the expression of viral nucleic acids in the lung tissue of the mice was significantly increased， with significant differences compared with the normal group （P<0.01）. The expression of viral nucleic acids in the ribavirin group and the high and low dose groups of Lianhua Qingwen was significantly reduced， with significant differences compared with the normal group （P<0.01）. ConclusionHumanized IGF1R mice are more susceptible to respiratory SVC， and the animal model of RSV-infected pneumonia based on humanized IGF1R mice was successfully constructed， which is suitable for the evaluation of anti-RSV drugs.

ObjectiveTo investigate the possible mechanism of Shufeng Jiedu capsules （SFJD） in alleviating influenza A （H1N1） virus pneumonia and focus on its effect on Toll-like receptor （TLR） signaling pathway in respiratory epithelial cells. MethodsA mouse model of viral pneumonia was established via the A/PR/8/34 （PR8） strain of influenza A virus. Mice were randomly divided into a normal group， a PR8 infection （PR8） group， and an SFJD group （8.4 g·kg^-1）， with 10 mice in each group. The day of infection was designated as day 1. The SFJD group was administered intragastrically at a volume of 20 mL·kg^-1 daily， while the normal and PR8 groups were given an equal volume of deionized water. Micro-computed tomography （Micro-CT） was performed on day 5， and the mice were dissected to collect their lungs， after which the lung index was calculated to verify the therapeutic effect of SFJD. Single-cell sequencing was used to analyze the differentially expressed genes in respiratory epithelial cells. Multiplex fluorescence immunohistochemistry was employed to detect the expression of TLR， tumor necrosis factor receptor-associated factor 6 （TRAF6）， and myeloid differentiation factor 88 （MyD88） proteins in epithelial cell adhesion molecule （EpCAM）-positive cells， and the proportion of respiratory epithelial cells expressing TLR pathway proteins was calculated. Respiratory epithelial cells were then sorted by flow cytometry， and Western blot was used to detect the expression of TLR， MyD88， TRAF6， Toll-interleukin receptor domain-containing adaptor inducing interferon-β （TRIF）， inhibitor of κB kinase α （IKKα）， and nuclear factor-κB （NF-κB） in the sorted epithelial cells. Enzyme-linked immunosorbent assay （ELISA） was used to measure the levels of interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） in lung tissue. ResultsAt the transcriptional level， SFJD reversed the expression of TLR signaling pathway genes in respiratory epithelial cells， downregulating multiple TLR signaling pathway-related genes （P<0.01）. At the protein level， SFJD significantly reduced the proportion of respiratory epithelial cells expressing TLR3 （P<0.05）， the expression levels of TLR2， TLR3， TLR4， TRIF， TRAF6， IKKα， and NF-κB in epithelial cells（P<0.05， P<0.01）， as well as the levels of pro-inflammatory cytokines IL-1β and TNF-α in lung tissue （P<0.01）. ConclusionSFJD may alleviate viral pneumonia by suppressing the expression of TLR in respiratory epithelial cells and their subsequent signaling cascades.

AIM To study the chemical constituents from the buds of Aralia chinensis L.var.nuda Nakai and their in vitro anti-inflammatory activities.METHODS The 70％ethanol extract from the buds of A.chinensis var.nuda was isolated and purified by silica gel,Sephadex LH-20,ODS and semi-preparative HPLC,then the structures of compounds were identified by physicochemical properties and spectral data.Their anti-inflammatory activities in vitro were evaluated by RAW264.7 model.RESULTS Sixteen compounds were isolated and identified as 4-(2,2-dibutoxyethyl)phenol(1),trans-linalool-3,7-oxide-6-O-β-D-glucopyranoside(2),2'-O-(9Z,12Z,15Z-octadecatrienoyl)glyceryl β-D-galactopyranoside(3),quercetin-3-O-β-D-glucopyranoside(3'→ O-3''')quercetin-3-O-β-D-galactopyranoside(4),syringaresinol-4'-O-β-D-glucopyranoside(5),p-hydroxybenzaldehyde(6),7α-hydroxystigmasterol 3-O-β-D-glucopyranoside(7),trans-p-hydroxy cinnamic acid methyl ester(8),funingensin A(9),3,4-dihydroxy-acetophenone(10),N-acetyltyramine(11),3,4-di-O-caffeoyl quinic acid(12),chlorogenic acid(13),aralia cerebroside(14),caffeic acid methyl ester(15),tetradecanoic acid(16).The IC50values of compounds 8,10,12 and 13 were(22.19±1.59),(35.25±1.30),(13.38±0.72),(15.73±1.16)μmol/L,respectively.CONCLUSION Compound 1 is a new compound,2-13 are isolated from genus Aralia for the first time.Compounds 8,10,12,13 exhibit significant in vitro anti-inflammatory activities.

Objective To investigate the expression of non-receptor protein tyrosine phosphatase 18(PT-PN18)and RNA binding protein FOX2(RBFOX2)in endometrial cancer(EC),and to analyze their relation-ship with clinicopathological features and prognostic significance.Methods A total of 122 EC patients treated in Hainan Women and Children's Medical Center from January 2020 to June 2021 were collected.The expres-sion of PTPN18 and RBFOX2 protein was detected by immunohistochemistry.The expressions of PTPN18 mRNA and RBFOX2 mRNA were detected by real-time fluorescence quantitative PCR.Pearson correlation a-nalysis was used to analyze the correlation between PTPN18 mRNA and RBFOX2 mRNA expression in EC.Kaplan-Meier curve and multivariate Cox regression model were used to analyze the effect of PTPN18 mRNA and RBFOX2 mRNA expression on the prognosis of EC patients.Results Compared with adjacent tissues,PTPN18 protein expression(2.53±0.67 vs.0.56±0.14)was higher and RBFOX2 protein expression(0.74±0.22 vs.3.11±0.63)was lower in EC cancer tissues,and the difference was statistically significant(t=31.790,39.229,P<0.001).Compared with adjacent tissues,PTPN18 mRNA expression(3.15±0.76 vs.0.78±0.23)was higher and RBFOX2 mRNA expression(1.14±0.34 vs.3.87±0.80)was lower in EC cancer tissues,and the difference was statistically significant(t=32.976,34.689,P<0.001).There was a negative correlation between the expression of PTPN18 mRNA and RBFOX2 mRNA in EC(r=-0.674,P<0.001).The expression of PTPN18 mRNA was higher and RBFOX2 mRNA was lower in FIGO stage Ⅲ EC patients with lymph node metastasis(P<0.05).There was no significant difference in 3-year overall survival rate between PTPN18 mRNA high expression group and low expression group,and between RBFOX2 mRNA high expression group and low expression group(P>0.05).The 3-year progression-free survival rate of the PTPN18 mRNA high expression group was lower than that of the low expression group,and the 3-year pro-gression-free survival rate of the RBFOX2 mRNA high expression group was higher than that of the low ex-pression group,and the differences were statistically significant(P<0.001).FIGO stage Ⅲ,lymph node me-tastasis and PTPN18 mRNA were risk factors for progression-free survival in EC patients,and RBFOX2 mR-NA was a protective factor(P<0.05).Conclusion PTPN18 expression is increased and RBFOX2 expression is decreased in EC,which is related to FIGO stage and lymph node metastasis.PTPN18 is a tumor marker for evaluating the prognosis of EC.

Objective To analyze the correlation between tongue image features and the risk levels of disease in patients with idiopathic membranous nephropathy(IMN).Methods Based on IMN clinical research electronic data acquisition system,a cross-sectional study method was used to analyze the clinical diagnosis and treatment data of 135 IMN patients from Guangdong Provincial Hospital of Chinese Medicine.The patients were grouped according to the risk levels of disease,and then the correlation between the risk levels of disease and tongue image features was analyzed.During the description of tongue image features,TB is for tongue body,TC is for tongue coating,L is for luminance,a is for red-green axis,G is for the value of green,B is for the value of blue,and AUT is for the value of autocorrelation.Results The comparison of tongue image feature indicators of patients with different risk levels of IMN showed that:(1)the higher the level of disease risk of IMN patients,the greater the values of TB-L,TB-G and TB-B(P<0.05 or P<0.01).The values of tongue image indicator TB-a and TC-a of the patients with different risk levels of IMN were shown in decreasing sequence:low-risk group>high-risk group>middle-risk group>extremely-high-risk group(P<0.05).(2)Linear regression analysis showed that TB-L,TB-G,and TB-B were significantly increased in the high-risk group compared with those in the middle-and low-risk groups(P<0.05 or P<0.01),whereas there were no significant differences between the middle-risk group and low-risk group(P>0.05).(3)The results of correlation analysis showed that there was a positive correlation among most of the tongue image feature indicators(including TB-L,TB-G,TB-B,TB-AUT,TC-L,TC-G,and TC-B,etc.)and the risk level of disease,while TB-a was negatively correlated with the risk level of disease,and the differences were all statistically significant(P<0.05 or P<0.01).(4)All patients were treated with Chinese medicine and/or Chinese patent medicine,and 46.7%of patients were given hormones and immunosuppressants,and there was no statistically significant difference in the the use of hormones and immunosuppressants among various groups(P=0.637).Conclusion There is a correlation between the tongue image features of IMN patients and the risk level of disease,and the results will provide an objective reference for the assessment of illness state and traditional Chinese medicine(TCM)syndrome differentiation of IMN patients.With reference to the changes in the tongue image features,the illness state can be precisely identified,which is more accurate than the inspection of four diagnostic methods of TCM.

Objective: This study aims to investigate the therapeutic effects and underlying mechanisms of Xuanfei Baidu Decoction (XFBD) in a mouse model of dampness-heat toxin pneumonia. By exploring how XFBD exerts its effects, we seek to deepen our understanding of its role in treating pulmonary diseases and to address the current knowledge gap regarding its mechanisms of action, thereby supporting its clinical application. Methods: Ultra-high-performance liquid chromatography and high-resolution mass spectrometry (HRMS) were employed to analyze the chemical constituents of XFBD. The protective effects of XFBD were evaluated using a dampness-heat toxin-induced mouse model, established through dampness-heat exposure and HCoV-229E infection. XFBD was administered orally, followed by assessments including lung index measurement, micro-CT imaging, viral load quantification, cytokine analysis, and histological evaluation via hematoxylin-eosin staining. Proteomics and single-cell transcriptomic analyses were conducted to explore the potential mechanisms underlying XFBD’s pharmacological effects. A cellular model of HCoV-229E infection was developed to investigate changes in the cAMP/PKA signaling pathway. Molecular docking and surface plasmon resonance (SPR) experiments confirmed the strong binding affinity between key XFBD components and PKA. Finally, PKA activators and inhibitors were applied in vitro to validate these mechanistic findings. Results: In vivo studies demonstrated that XFBD significantly reduced the lung index, improved the structural integrity of lung and tongue tissues, and decreased levels of proinflammatory mediators, including IL-6, IL-8, and TNF-α. Proteomic and single-cell transcriptomic analyses showed that the differentially expressed proteins after XFBD treatment were primarily associated with inflammatory responses and immune regulation. The cAMP/PKA signaling pathway was identified as a key mechanism underlying these therapeutic effects. Notably, Western blot, ELISA, molecular docking, and SPR analyses confirmed that XFBD elevated cAMP levels and p-PKA expression, thereby activating the cAMP/PKA signaling pathway in vitro. Conclusion: This study demonstrated that XFBD significantly alleviates symptoms in mice with dampness-heat toxin pneumonia. Its therapeutic effects are mediated, at least in part, through activation of the cAMP/PKA signaling pathway. These findings provide compelling evidence that XFBD is an effective herbal remedy against HCoV-229E infection.

Objective:To evaluate the characteristic manifestations of lumbar fusion following oblique lateral interbody fusion (OLIF), determine the specific incidence of these patterns, and the identify factors associated with fusion characteristics.Methods:This retrospective study analyzed 209 patients who underwent OLIF surgery at Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, between July 2017 and September 2023. The cohort had a mean age of 64.2±9.8 years and included 125 males and 84 females. A total of 338 lumbar segments were assessed, comprising 159 segments treated with stand-alone OLIF (OLIF-SA) and 179 segments treated with OLIF combined with posterior pedicle screw fixation (OLIF-PSF). Inclusion criteria were: patients aged 18-80 years who underwent OLIF with complete radiographic records. Surgical parameters, including fixation method, number of fused segments, surgical approach, and cage dimensions (height and width), were obtained from operative records. Radiographic evaluation included preoperative osteophytes, Hounsfield unit (HU) values of endplates, and cage positioning. Fusion rate, fusion pattern (lateral vs. central), cage subsidence, and related influencing factors were assessed. Clinical outcomes were measured via the Oswestry disability index (ODI) and visual analog scale (VAS) preoperatively, immediately postoperatively, and at 1-year follow-up. Results:The overall fusion rate was 98.2% (332/338), with a non-union rate of 1.8% (6/338). The incidence of lateral fusion was 40.2% (136/338). In the OLIF-SA group, lateral and central fusion rates were 50.3% (80/159) and 49.7% (79/159), respectively, with no cases of non-union. In the OLIF-PSF group, lateral fusion occurred in 31.3% (56/179), central fusion in 65.4% (117/179), and non-union in 3.3% (6/179), with statistically significant differences between groups ( P<0.05). Preoperative osteophytes and higher endplate HU values were significantly associated with lateral fusion ( P<0.05). However, cage dimensions and cage position (anterior-posterior and lateral placement) were not significantly associated with fusion pattern ( P>0.05). Overall, 61.5% (208/338) of segments showed no cage subsidence; 24.5% (83/338) had settling, and 14.0% (47/338) had grade 1 or higher subsidence. Among lateral fusion cases, the rates of no subsidence, anchoring, grade 1, grade 2, and grade 3 subsidence were 67.6%, 21.3%, 7.4%, 3.9%, and 0.7%, respectively. In the central fusion group, these rates were 59.2%, 27.6%, 9.2%, 2.5%, and 1.3%, respectively. In the non-union group, grade 2 and 3 subsidence occurred in 50% (3/6) each, significantly higher than in the other fusion groups ( P<0.05). Post hoc analysis confirmed that grade 2 and 3 subsidence rates were significantly elevated in the non-union group compared to the lateral and central fusion groups, while other subsidence categories showed no significant differences across groups. Clinically, patients showed significant improvements in ODI and VAS scores following surgery ( P<0.05). Conclusions:Lateral fusion occurred in 40.2% of OLIF cases. The OLIF-SA technique, preoperative osteophytes, and elevated preoperative HU values were significantly associated with lateral fusion. In contrast, surgical approach, number of fused segments, cage height, width, and cage positioning did not significantly influence the occurrence of lateral fusion.

Objective To systematically evaluate the research progress of risk prediction models for periprosthetic joint infection(PJI)after total joint arthroplasty(TJA),analyze the limitations of current researches,and propose optimized suggestions.Methods Chinese and English databases such as PubMed,Embase,Web of Science,Co-chrane Library,SinoMed,Wanfang Database,VIP Database,and CNKI were retrieved systematically.The re-trieved period was from the establishment of each database to August 31,2024.Two researchers independently screened literatures and extracted data according to the CHARMS checklist,and the risk of bias in the included studies was evaluated by the PROBAST tool.Results A total of 14 studies were included in this study,involving 17 prediction models.The most common predictors included history of diabetes mellitus,obesity(body mass index[BMI]≥30 kg/m2),advanced age(≥65 years old),history of traumatic fracture,and prolonged operation time(≥2 hours).All of the included studies had high risks of bias,mainly study subject selection bias(such as single-center sample)and statistical analysis bias(such as unadjusted confounding factors).Conclusion Most of the cur-rently published risk prediction models for PJA after TJA have good predictive performance,however,there are sig-nificant limitations in the research design,especially in the insufficient control of bias risk.Future research needs to focus on improving methodological design,including adoption of prospective multi-center studies,definition of standardized predictive variables,and sufficient adjustment of confounding factors.

Hypokalemia,a common clinical electrolyte disorder,can affect multiple systems and can be life-threatening in severe cases.Identifying the cause of hypokalemia is crucial for its prevention and treatment.However,the etiology of hypokalemia is complex and often requires detailed differential diagnosis.This article reports a rare clinical case of hypokalemia caused by long-term excessive consumption of strong tea and discusses its pathogenesis.The aim is to raise clinical awareness and understanding of the etiology of such cases of hypokalemia and reduce misdiagnosis and missed diagnosis.