ObjectiveTo establish an in vitro culture model of rat corpora cavernous smooth muscle cells （CCSMCs） using a modified tissue block adherence method. MethodsCorpus cavernosum smooth muscle tissue was digested with collagenase type I and subsequently cultured using an adherent method. Cells were purified via differential adhesion and identified through immunofluorescence and Western blotting. ResultsCCSMCs began to emerge from the tissue block after 3 days， increased significantly by day 7， and converged by day 12. Post-passage， CCSMCs exhibited strong proliferation and a “peak-to-valley” phenomenon. After purification， the cells tested positive for α-smooth muscle actin （α-SMA）， confirming the successful establishment of the in vitro culture model. ConclusionThe modified tissue block adherence method is a cost-effective and efficient way to obtain high-purity CCSMCs.

BACKGROUND:Spinal canal decompression using uniportal endoscopic surgery is a new minimally invasive surgery in the treatment of lumbar spinal stenosis.However,this technique needs a steep learning curve and high requirements for surgical equipment and instruments,which limits its clinical application.We previously use the spinal endoscopy as a monitoring endoscopy and combined with unilateral biportal endoscopy to propose a hybrid technique of spinal endoscopy to achieve coaxial endoscopic operation and hands-separate operation. OBJECTIVE:To compare the clinical outcome of hybrid technique and uniportal endoscopic surgery in treatment of lumbar spinal stenosis with bilateral lower limb pain symptoms. METHODS:Ninety patients diagnosed of lumbar spinal stenosis with bilateral symptoms were included and retrospectively analyzed at First People's Hospital,Shanghai Jiao Tong University from August 2020 to August 2022.44 cases were included in group A(hybrid technique group),while 46 cases were included in group B(uniportal endoscopic surgery).The nerve decompression was observed during the surgery.Operation time,hospital stay time,and expenses were recorded in both groups.The visual analog scale scores of lower back pain and both lower extremities pain,Oswestry disability index scores of quality of life and excellent and good rate of modified Macnab criteria were recorded and compared at preoperative,postoperative 3 days,and postoperative 3 and 6 months. RESULTS AND CONCLUSION:(1)The operation time of group A was significantly shorter than that of group B(P<0.05).(2)The lower back pain and lower extremity pain of the severe side at postoperative 3 days,and 3 and 6 months were significantly relieved in both groups(P<0.05).The visual analog scale score of lower extremity pain on the mild side was significantly decreased at postoperative 3 days,3 and 6 months than preoperative score in the group A(P<0.05).The visual analog scale score of lower extremity pain on the mild side was significantly decreased at postoperative 3 days than preoperative score in the group B(P<0.05).The visual analog scale scores of lower extremity pain on the mild side at postoperative 3 and 6 months did not show significant difference than preoperative score in the group B.The comparison between the two groups showed that there was no significant difference in the visual analog scale scores of postoperative lower back pain and lower extremity pain of the severe side(P>0.05).The visual analog scale scores of lower extremity pain on the mild side in the group A were significantly lower than those of group B at postoperative 3 and 6 months(P<0.05).(3)The Oswestry disability index scores of both groups at postoperative 3 day were significantly lower than preoperative score(P<0.05),and there was no significant difference between the two groups 3 days after operation.Oswestry disability index scores of group A at postoperative 3 and 6 months were significantly decreased than preoperative score(P<0.05).The Oswestry disability index scores of group B at postoperative 3 and 6 months did not show significant differences than preoperative score(P>0.05).The comparison between the two groups showed the Oswestry disability index scores of group A were significantly lower than group B at postoperative 3 and 6 months(P<0.05).(4)The results of modified Macnab showed that the excellent and good rate of group A was significantly higher than that of group B(95%,78%,P<0.05).(5)It is indicated that the hybrid technique is a new spinal endoscopy technique,which has the advantages of less trauma and faster recovery as a minimally invasive surgery.The clinical outcome of hybrid technique is superior to that of uniportal endoscopic surgery in the treatment of lumbar spinal stenosis with bilateral symptoms.Additionally,it also has the advantages of good operational flexibility and high decompression efficiency as an open surgery.

Dental mesenchymal stem cells (DMSCs) are pivotal for tooth development and periodontal tissue health and play an important role in tissue engineering and regenerative medicine because of their multidirectional differentiation potential and self-renewal ability. The cellular microenvironment regulates the fate of stem cells and can be modified using various optimization techniques. These methods can influence the cellular microenvironment, activate disparate signaling pathways, and induce different biological effects. "Epigenetic regulation" refers to the process of influencing gene expression and regulating cell fate without altering DNA sequences, such as histone methylation. Histone methylation modifications regulate pivotal transcription factors governing DMSCs differentiation into osteo-/odontogenic lineages. The most important sites of histone methylation in tooth organization were found to be H3K4, H3K9, and H3K27. Histone methylation affects gene expression and regulates stem cell differentiation by maintaining a delicate balance between major trimethylation sites, generating distinct chromatin structures associated with specific downstream transcriptional states. Several crucial signaling pathways associated with osteogenic differentiation are susceptible to modulation via histone methylation modifications. A deeper understanding of the regulatory mechanisms governing histone methylation modifications in osteo-/odontogenic differentiation and immune-inflammatory responses of DMSCs will facilitate further investigation of the epigenetic regulation of histone methylation in DMSC-mediated tissue regeneration and inflammation. Here is a concise overview of the pivotal functions of epigenetic histone methylation at H3K4, H3K9, and H3K27 in the regulation of osteo-/odontogenic differentiation and renewal of DMSCs in both non-inflammatory and inflammatory microenvironments. This review summarizes the current research on these processes in the context of tissue regeneration and therapeutic interventions.
Mesenchymal Stem Cells/physiology* ; Humans ; Osteogenesis/genetics* ; Histones/metabolism* ; Cell Differentiation/physiology* ; Methylation ; Odontogenesis/genetics* ; Epigenesis, Genetic

Flavonoids, the key active compounds in Epimedii Folium, have both protective and toxic effects on the liver. Their hepatoprotective effects are associated with reducing lipid accumulation and oxidative stress, which contribute to the management of various liver conditions. In contrast, the mechanisms driving Epimedii Folium-induced hepatotoxicity are less understood but likely involve oxidative stress and pyroptosis. Pharmacokinetic studies, especially on icaritin, indicate that it undergoes isopentenyl dehydrogenation, glycosylation, and glucuronidation in vivo, contributing to its pharmacological effects. However, intermediate metabolites of icaritin may interact with biomolecules, potentially leading to liver toxicity. This review offers a detailed examination of the dual effects of Epimedii Folium flavonoids on liver function, emphasizing recent discoveries in their hepatoprotective and hepatotoxic pathways. We also summarize and discuss the pharmacokinetics of these flavonoids, highlighting how their metabolism affects therapeutic efficacy and toxicity. Lastly, we propose strategies to mitigate liver injury, providing new perspectives on the safe use of Epimedii Folium.

A comprehensive phytochemical investigation of the leaves and twigs of Physalis angulata. var. villosa resulted in the isolation of 23 withanolide derivatives, including one novel 13,20-γ-lactone withanolide derivative (1) and three new withanolide derivatives (2-4). Architecturally, physalinin A (1) represents the first identified type B withanolide featuring a 13,20-γ-lactone moiety. The molecular structures of all isolates were elucidated using an integrated approach combining nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry (MS), infrared (IR) spectroscopy, and quantum chemical calculations to confirm structural assignments. The antiproliferative activities of all isolated withanolides were evaluated against four human cancer cell lines (HEL, HCT-116, Colo320DM, and MDA-MB-231). Among them, eight derivatives (2, 5-8, 14, 15, and 23) exhibited significant inhibitory effects, with half-maximal inhibitory concentration (IC₅₀) values of 0.18 ± 0.03 to 17.02 ± 0.21 μmol·L^-1. Structure-activity relationship (SAR) analysis suggested that the presence of an epoxide ring enhances anticancer activity, potentially through increased reactivity or specific interactions with molecular targets involved in cancer progression. These findings underscore the pharmacological potential of withanolides as promising lead compounds for the development of novel anticancer therapeutics.
Withanolides/isolation & purification* ; Physalis/chemistry* ; Humans ; Molecular Structure ; Cell Line, Tumor ; Antineoplastic Agents, Phytogenic/isolation & purification* ; Cell Proliferation/drug effects* ; Plant Leaves/chemistry* ; Plant Extracts/pharmacology*

Objective To analyze the expression levels and the clinical significance of serum interleukin(IL)-4,IL-5,IL-6,IL-13 and IL-17 in children with mycoplasma pneumoniae(MP)infection accompanied by airway hyperresponsiveness(AHR).Methods A total of 120 children diagnosed with MP infection pneumonia and admitted to Nantong First People's Hospital between June 2022 and April 2024 were enrolled in the study group.According to whether their MP infection pneumonia was accompanied by AHR,the participants were divided into an AHR group(n=41)and a non-AHR group(n=79).An additional 90 healthy children undergoing physical examination were included in the control group.Baseline data,fractional exhaled nitric oxide(FeNO),tidal breathing lung function—including the ratio of volume at peak tidal expiratory flow to total expiratory volume(VPTEF/VE),the ratio of time to peak tidal expiratory flow to total expiratory time(TPTEF/TE),and inspiratory-to-expiratory time ratio(TI/TE)—as well as the levels of serum IL-4,IL-5,IL-6,IL-13 and IL-17 were compared among the 3 groups.The correlation between serum IL-4,IL-5,IL-6,IL-13 and IL-17 and clinical data of children with MP infection pneumonia accompanied by AHR was analyzed.Logistic regression analysis was used to identify the independent influencing factors of AHR in children with MP infection pneumonia.Receiver operating characteristic(ROC)curves were plotted to assess the diagnostic value of serum IL-4,IL-5,IL-6,IL-13 and IL-17 for MP infection pneumonia accompanied by AHR in children.Results Significant differences were observed among the AHR,non-AHR,and control groups in white blood cell(WBC)count,neutrophil percentage,eosinophil(EOS)count,platelet(PLT)count,C-reactive protein(CRP),FeNO,VPTEF/VE,TPTEF/TE,TI/TE,forced vital capacity(FVC),and forced expiratory volume in one second(FEV?)(all P<0.05).Serum IL-4,IL-5,IL-6,IL-13 and IL-17 levels showed significant differences among the 3 groups(P<0.05).Correlation analysis showed that these cytokines were positively correlated with WBC count,neutrophil percentage,EOS count,CRP,and FeNO(P<0.05),and negatively correlated with VPTEF/VE,TPTEF/TE,TI/TE,FVC,and FEV1(P<0.05).Binary logistic regression analysis identified neutrophil percentage(odds ratio[OR]=1.923;95%CI,1.496-2.472),EOS count(OR=3.074;95%CI,1.228-7.693),CRP(OR=2.382;95%CI,1.854-3.061),FeNO(OR=1.931;95%CI,1.635-2.281),VPTEF/VE(OR=0.294;95%CI,0.200-0.432),TPTEF/TE(OR=0.358;95%CI,0.177-0.722),TI/TE(OR=0.399;95%CI,0.221-0.722),IL-4(OR=1.064;95%CI,1.019-1.111),IL-5(OR=1.234;95%CI,1.095-1.390),IL-6(OR=1.013;95%CI,1.001-1.025),IL-13(OR=1.058;95%CI,1.005-1.113),IL-17(OR=1.759;95%CI,1.293-2.393),and allergy history(OR=2.989;95%CI,1.058-8.447)as independent factors associated with with AHR in children with MP infection pneumonia(P<0.05).ROC curves revealed that the areas under the ROC curves(AUC)of serum IL-4,IL-5,IL-6,IL-13 and IL-17 for predicting AHR in children with MP infection pneumonia were 0.815,0.769,0.782,0.793,and 0.815,respectively,with the sensitivities being 85.37%,75.61%,87.80%,75.61%,and 80.49%,and 95%CI being 0.739-0.892,0.677-0.861,0.679-0.867,0.715-0.882 and 0.732-0.899,respectively.Conclusion The levels of serum IL-4,IL-5,IL-6,IL-13,and IL-17 in children with MP infection pneumonia and AHR are abnormally elevated.These cytokines may serve as valuable biomarkers for assessing the risk of AHR in children with MP infection pneumonia.

Objective To predict and preliminarily validate potential shared key genes involved in cardiac complications caused by influenza A virus(IAV)and severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infections.Methods Differentially expressed genes(DEGs)associated with cardiac complications were obtained from the Gene Expression Omnibus(GEO)database.A hierarchical intersection strategy was applied.First,cardiac complication related DEGs were overlapped with 2 independent virus related gene sets:3 454 human genes linked to IAV infection in GeneCards and 333 human protein-coding genes interacting with SARS-CoV-2 in the Human Protein Atlas.The 2 overlap results were then intersected to yield 22 hub genes.Lasso regression,random forest(RF)and support vector machine algorithms(SVM)were employed to refine this list.Predicted genes were validated in vitro in H1N1-infected human cardiomyocyte AC16 cells and in vivo in IFITM3 knockout mice challenged with H1N1,assessing transcriptional changes.Results A total of 22 hub genes were identified through integrative bioinformatics analysis.Application of the 3 machine learning algorithms resulted in 5 common key genes:ACE2,TBK1,NUP210,PUSL1,and MEPCE.In vitro infection of AC16 cells with H1N1 revealed dynamic transcriptional changes in all 5 genes post-infection(P<0.05).In vivo experiments using H1N1-infected IFITM3 knockout mice confirmed the dynamic mRNA expression changes of these 5 genes,consistent with the in vitro results(P<0.05).Conclusion By combining multilayered bioinformatics analysis with 3 machine learning approaches,5 common key genes are identified:ACE2,TBK1,NUP210,PUSL1 and MEPCE.Validation in H1N1 infection models confirms their relevance to IAV-induced cardiac complications.

Objective To explore the clinical value of serum cysteine-rich protein 61(CYR61)and cardiac fatty acid-binding protein(H-FABP)in the diagnosis of neonatal acute respiratory distress syndrome.Methods A total of 105 children with acute respiratory distress syndrome who received treatment in the hos-pital from November 2020 to November 2022 were selected as the study group,and divided into mild group(42 cases),moderate group(35 cases)and severe group(28 cases).In addition,60 healthy newborns in the same period were selected as the control group.Serum CYR61 and H-FABP levels were detected and com-pared in all subjects after admission.Receiver operating characteristic(ROC)curve and area under curve(AUC)were used to evaluate the diagnostic value of serum CYR61 and H-FABP in neonatal acute respiratory distress syndrome.The related factors affecting the occurrence of neonatal acute respiratory distress syndrome were explored by multivariate Logistic regression.Results The levels of serum CYR61 and H-FABP in the study group were higher than those in the control group,and the differences were statistically significant(P＜0.05).Serum CYR61 and H-FABP levels in severe group were higher than those in moderate and mild groups(severe group＞moderate group＞mild group),and the differences were statistically significant(P＜0.05).ROC curve analysis showed that the AUC of serum CYR61 for neonatal acute respiratory distress syndrome was 0.843(95％CI:0.824-0.893).The AUC of serum H-FABP for neonatal acute respiratory distress syn-drome was 0.864(95％CI:0.814-0.914).The AUC of the combined detection for neonatal acute respiratory distress syndrome were 0.925(95％CI:0.875-0.975).Multivariate Logistic stepwise regression analysis showed that serum CYR61(OR=3.050,95％CI:1.738-5.352),H-FABP(OR=3.773,95％CI:1.845-7.717),C-reactive protein(OR=2.349,95％CI:1.584-3.483)and oxygenation index(OR=1.944,95％CI:1.444-2.619)were risk factors for neonatal acute respiratory distress syndrome(P＜0.05).Conclusion Ser-um CYR61 and H-FABP are both elevated in neonatal acute respiratory distress syndrome,and are closely re-lated to the severity of the disease,which are expected to be effective biological indexes for early diagnosis of neonatal acute respiratory distress syndrome.

Objective To explore the clinical efficacy and safety of donepezil(DNPQ)combined with butylphthalide sequential therapy(BST)in the treatment of Parkinson's syndrome(PS).Methods In this study,104 patients with Parkinson's disease(PD)who were diagnosed and treated in the Department of Neurology of The Fourth Hospital of Changsha from January 2020 to November 2023 were randomly divided into a control group(butylphthalide softcapsule combined with DNPQ)and an observation group(BST combined with DNPQ).The main observation indicators of this study were the clinical efficacy and drug-related adverse reactions after 3 months of treatment.The secondary observation indicators were the cognitive function[Montreal Cognitive Assessment(MoCA)and Mini-Mental State Examination(MMSE)],overall condition[Unified Parkinson's Disease Rating Scale(UPDRS)],activity of daily living(ADL),and oxidative stress-related cytokines[recombinant human Parkinson's disease protein 7(PARK7),neurotrophic factor 3(NT3),and C-reactive protein(CRP)]improvement after treatment.Results There were 52 patients in the experimental group and 52 patients in the control group.The treatment efficacy rate in the experimental group was significantly higher than that in the control group(P<0.05),while the incidence of adverse reactions was significantly lower than that in the control group(P<0.05).Before treatment,there were no significant differences in MoCA scores,MMSE scores,UPDRS scores,ADL scores,serum NT3,CRP,and PARK7 levels between the two groups(P>0.05).After treatment,the MoCA score,MMSE score,and ADL score in the experimental group were higher than those in the control group(P<0.05),while the UPDRS score was lower than that in the control group(P<0.05).After treatment,the serum NT3 level in the experimental group was higher than that in the control group(P<0.05),while the serum CRP and PARK7 levels were lower than those in the control group(P<0.05).Conclusion The combination of DNPQ and BST has better clinical efficacy and safety,which can improve cognitive function,ADL and oxidative stress-related cytokine content in patients with PS.

Objective:To explore the current situation and influencing factors of amputation decision-making dilemma of diabetic foot patients.Methods:From July to December 2023, 200 patients with diabetic foot in the Tianjin Medical University General Hospital and Tianjin Medical University Chu Hsien-I Memorial Hospital were selected as study subjects by convenience sampling. General Information Questionnaire, Decisional Conflict Scale (DCS), Family APGAR Index, and Hospital Anxiety and Depression Scale were used to conduct a cross-sectional survey. Pearson correlation was used to analyze the correlation between diabetic foot patients' amputation decision-making dilemma and family caring, anxiety and depression, and multiple linear regression was used to analyze the influencing factors of diabetic foot patients' amputation decision-making dilemma.Results:A total of 200 questionnaires were distributed, and 180 valid questionnaires were collected, with a valid response rate of 90.0% (180/200). The DCS score of 180 patients with diabetic foot was (30.04±9.77), 76.7% (138/180) patients scored ≥25.0, and they had decision-making dilemma, and 25.0% (45/180) of patients scored ≥37.5, indicating decision-making delay. Multiple linear regression analysis showed that occupational status, diabetes course, family caring, anxiety and depression were the influencing factors of amputation decision-making dilemma of diabetic foot patients ( P<0.05) . Conclusions:Diabetic foot patients face certain dilemmas in the process of amputation decision-making. Clinical medical and nursing staff should reasonably evaluate the patient's occupational status, disease course, family caring, and psychological state, and develop personalized decision support strategies to improve decision quality and prevent changes in the patient's condition caused by delayed decision-making.