Objective To investigate the effect and underlying mechanism of bufalin regulating ferroptosis on extracellular matrix synthesis in renal tubular epithelial cells(RTECs)under high glucose(HG)conditions.Methods RTECs were cultured in vitro and exposed to HG.The experimental groups included:the control group,the HG group,the HG+dimethyl sulfoxide(DMSO)group,the HG+bufalin group,the HG+ferrostatin-1(Fer-1)group,the HG+bufalin+DMSO group and HG+bufalin+erastin group.The expression levels of fibronectin(FN),type Ⅰ collagen(Col Ⅰ),acyl-CoA synthetase long-chain family member 4(ACSL4),solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4)were detected using Western blot assay and quantitative real-time PCR(qRT-PCR).The potential molecular targets of bufalin were predicted using SwissTargetPrediction,and functional enrichment analysis was conducted using Metascape.FerrDb was employed to analyze ferroptosis-related gene sets.The levels of ferrous ions(Fe2+),malondialdehyde(MDA)and glutathione(GSH)were measured using micro-methods to evaluate the occurrence of ferroptosis.Results Compared with the control group,the mRNA and protein relative expression levels of FN,Col Ⅰand ACSL4 were increased in the HG group,while the mRNA and protein expression levels of GPX4 and SLC7A11 were decreased(P＜0.05).Compared with the HG+DMSO group,the mRNA and protein expression levels of FN,Col Ⅰand ACSL4,as well as levels of Fe2+and MDA were decreased in the HG+bufalin group,while the mRNA and protein expression levels of GPX4 and SLC7A11,and the level of GSH were increased(P＜0.05).In the HG+Fer-1 group,the mRNA and protein expression levels of GPX4 and SLC7A11 were increased,while the mRNA and protein expression levels of ACSL4,FN and Col Ⅰ were decreased(P＜0.05).The SwissTargetPrediction database and Metascape analysis function showed that the downstream functions of bufalin were closely related to lipid metabolism,inflammatory response,programmed cell death and ferroptosis-related pathways.The FerrDb analysis results indicated that the target sites of bufalin were closely related to ferroptosis markers.Compared with the HG+bufalin+DMSO group,the mRNA and protein expression levels of GPX4 and SLC7A11 were decreased in the HG+bufalin+Erastin group,while the mRNA and protein expression levels of ACSL4,FN and Col Ⅰ were increased(P＜0.05).Conclusion Bufalin attenuates extracellular matrix synthesis in HG-induced RTECs by inhibiting ferroptosis.

Objective To investigate the value of the combined detection of neutrophil CD64(nCD64),C-reactive protein(CRP)and lactate dehydrogenase(LDH)in the diagnosis of refractory Mycoplasma pneumoniae pneumonia(RMPP)in children.Methods A total of 147 children with Mycoplasma pneumoniae infection were enrolled and divided into the RMPP group(n=70)and the general Mycoplasma pneumoniae pneumonia group(GMPP,n=77)based on disease severity and treatment response.The age,gender,white blood cell count(WBC)within 24 hours of admission,serum procalcitonin(PCT),C-reactive protein(CRP)and lactate dehydrogenase(LDH)levels were collected in the study participants.The expression level of nCD64 in peripheral blood was measured using flow cytometry.Multivariate Logistic regression analysis was conducted to identify the independent risk factors associated with RMPP in children.Additionally,a receiver operating characteristic(ROC)curve was constructed to assess the diagnostic performance of the combined detection of nCD64,CRP and LDH for RMPP in children.Results The RMPP group had a longer hospital stay than the GMPP group(P＜0.05).Levels of nCD64,CRP and LDH were significantly higher in the RMPP group compared to those of the GMPP group(P＜0.05),and there were no significant differences in WBC and PCT levels between the two groups.Multivariate Logistic regression analysis showed that elevated nCD64,CRP and LDH were risk factors for RMPP in children(P＜0.05).ROC curve analysis revealed that the areas under the curve(AUC)for nCD64,CRP and LDH in diagnosing RMPP were 0.817,0.863 and 0.805,respectively.The combined detection of three indicators for AUC was 0.948.Conclusion The levels of nCD64,CRP and LDH in blood of children with RMPP are higher than those of children with GMPP.The combined detection of the three indicators has a high diagnostic value for RMPP in children with Mycoplasma pneumoniae pneumonia.

Objective:To summarize and analyze the risk factors associated with bronchial mucus plugs (BMPs) in children with Mycoplasma pneumoniae pneumonia (MPP),providing a clinical basis for the indication of bronchoscopy in children with MPP.Methods:A total of 72 children with MPP admitted to the Department of Pediatrics at the Fourth Affiliated Hospital of Nanjing Medical University from December 2022 to December 2024,who underwent bronchoscopy,were selected for this study.According to the presence or absence of BMPs under tracheoscopy,the patients were divided into the mucus plug group and the non-mucus plug group,and their clinical data were analyzed and compared.Results:Among the 72 children with MPP,30 cases (41.7%) were in the mucus plug group,while 42 cases (58.3%) were in the non-mucus plug group.Univariate Logistic regression analysis showed that thermal peak ( OR 2.086,95% CI 1.029-4.299, P=0.041),thermal duration ( OR 1.453,95% CI 1.197-1.763, P < 0.001),thermal duration ≥10 days ( OR 26.154,95% CI 5.316-128.661, P<0.001),combined with pleural effusion ( OR 4.727,95% CI 1.136-19.677, P=0.033),atelectasis ( OR 5.636,95% CI 2.024-15.699, P<0.001),lymphocyte proportion( OR 0.936,95% CI 0.888-0.978, P=0.014),neutrophil proportion( OR 1.048,95% CI 1.005-1.093, P=0.028),lactatedehydrogenase ( OR 1.005,95% CI 1.001-1.009, P = 0.028),D-dimer ( OR 2.203,95% CI 1.133-4.280, P = 0.020),fibrin degradation products ( OR 1.563,95% CI 1.095-2.231, P=0.014),and no heat regression within 72 hours of hormone use ( OR 0.239,95% CI 0.085-0.667, P=0.006) were significant influencing factors for the formation of BMPs in children with MPP.Multivariate Logistic regression analysis showed that thermal duration ≥10 days and atelectasis were independent risk factors for BMPs formation ( P<0.05). Conclusion:For MPP children with thermal duration≥10 days,especially with atelectasis,BMPs may have been formed,and it is recommended to perform bronchoscopy as soon as possible.

Objective To investigate the value of the combined detection of neutrophil CD64(nCD64),C-reactive protein(CRP)and lactate dehydrogenase(LDH)in the diagnosis of refractory Mycoplasma pneumoniae pneumonia(RMPP)in children.Methods A total of 147 children with Mycoplasma pneumoniae infection were enrolled and divided into the RMPP group(n=70)and the general Mycoplasma pneumoniae pneumonia group(GMPP,n=77)based on disease severity and treatment response.The age,gender,white blood cell count(WBC)within 24 hours of admission,serum procalcitonin(PCT),C-reactive protein(CRP)and lactate dehydrogenase(LDH)levels were collected in the study participants.The expression level of nCD64 in peripheral blood was measured using flow cytometry.Multivariate Logistic regression analysis was conducted to identify the independent risk factors associated with RMPP in children.Additionally,a receiver operating characteristic(ROC)curve was constructed to assess the diagnostic performance of the combined detection of nCD64,CRP and LDH for RMPP in children.Results The RMPP group had a longer hospital stay than the GMPP group(P＜0.05).Levels of nCD64,CRP and LDH were significantly higher in the RMPP group compared to those of the GMPP group(P＜0.05),and there were no significant differences in WBC and PCT levels between the two groups.Multivariate Logistic regression analysis showed that elevated nCD64,CRP and LDH were risk factors for RMPP in children(P＜0.05).ROC curve analysis revealed that the areas under the curve(AUC)for nCD64,CRP and LDH in diagnosing RMPP were 0.817,0.863 and 0.805,respectively.The combined detection of three indicators for AUC was 0.948.Conclusion The levels of nCD64,CRP and LDH in blood of children with RMPP are higher than those of children with GMPP.The combined detection of the three indicators has a high diagnostic value for RMPP in children with Mycoplasma pneumoniae pneumonia.

Objective To investigate the effect and underlying mechanism of bufalin regulating ferroptosis on extracellular matrix synthesis in renal tubular epithelial cells(RTECs)under high glucose(HG)conditions.Methods RTECs were cultured in vitro and exposed to HG.The experimental groups included:the control group,the HG group,the HG+dimethyl sulfoxide(DMSO)group,the HG+bufalin group,the HG+ferrostatin-1(Fer-1)group,the HG+bufalin+DMSO group and HG+bufalin+erastin group.The expression levels of fibronectin(FN),type Ⅰ collagen(Col Ⅰ),acyl-CoA synthetase long-chain family member 4(ACSL4),solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4)were detected using Western blot assay and quantitative real-time PCR(qRT-PCR).The potential molecular targets of bufalin were predicted using SwissTargetPrediction,and functional enrichment analysis was conducted using Metascape.FerrDb was employed to analyze ferroptosis-related gene sets.The levels of ferrous ions(Fe2+),malondialdehyde(MDA)and glutathione(GSH)were measured using micro-methods to evaluate the occurrence of ferroptosis.Results Compared with the control group,the mRNA and protein relative expression levels of FN,Col Ⅰand ACSL4 were increased in the HG group,while the mRNA and protein expression levels of GPX4 and SLC7A11 were decreased(P＜0.05).Compared with the HG+DMSO group,the mRNA and protein expression levels of FN,Col Ⅰand ACSL4,as well as levels of Fe2+and MDA were decreased in the HG+bufalin group,while the mRNA and protein expression levels of GPX4 and SLC7A11,and the level of GSH were increased(P＜0.05).In the HG+Fer-1 group,the mRNA and protein expression levels of GPX4 and SLC7A11 were increased,while the mRNA and protein expression levels of ACSL4,FN and Col Ⅰ were decreased(P＜0.05).The SwissTargetPrediction database and Metascape analysis function showed that the downstream functions of bufalin were closely related to lipid metabolism,inflammatory response,programmed cell death and ferroptosis-related pathways.The FerrDb analysis results indicated that the target sites of bufalin were closely related to ferroptosis markers.Compared with the HG+bufalin+DMSO group,the mRNA and protein expression levels of GPX4 and SLC7A11 were decreased in the HG+bufalin+Erastin group,while the mRNA and protein expression levels of ACSL4,FN and Col Ⅰ were increased(P＜0.05).Conclusion Bufalin attenuates extracellular matrix synthesis in HG-induced RTECs by inhibiting ferroptosis.

Objective:To summarize and analyze the risk factors associated with bronchial mucus plugs (BMPs) in children with Mycoplasma pneumoniae pneumonia (MPP),providing a clinical basis for the indication of bronchoscopy in children with MPP.Methods:A total of 72 children with MPP admitted to the Department of Pediatrics at the Fourth Affiliated Hospital of Nanjing Medical University from December 2022 to December 2024,who underwent bronchoscopy,were selected for this study.According to the presence or absence of BMPs under tracheoscopy,the patients were divided into the mucus plug group and the non-mucus plug group,and their clinical data were analyzed and compared.Results:Among the 72 children with MPP,30 cases (41.7%) were in the mucus plug group,while 42 cases (58.3%) were in the non-mucus plug group.Univariate Logistic regression analysis showed that thermal peak ( OR 2.086,95% CI 1.029-4.299, P=0.041),thermal duration ( OR 1.453,95% CI 1.197-1.763, P < 0.001),thermal duration ≥10 days ( OR 26.154,95% CI 5.316-128.661, P<0.001),combined with pleural effusion ( OR 4.727,95% CI 1.136-19.677, P=0.033),atelectasis ( OR 5.636,95% CI 2.024-15.699, P<0.001),lymphocyte proportion( OR 0.936,95% CI 0.888-0.978, P=0.014),neutrophil proportion( OR 1.048,95% CI 1.005-1.093, P=0.028),lactatedehydrogenase ( OR 1.005,95% CI 1.001-1.009, P = 0.028),D-dimer ( OR 2.203,95% CI 1.133-4.280, P = 0.020),fibrin degradation products ( OR 1.563,95% CI 1.095-2.231, P=0.014),and no heat regression within 72 hours of hormone use ( OR 0.239,95% CI 0.085-0.667, P=0.006) were significant influencing factors for the formation of BMPs in children with MPP.Multivariate Logistic regression analysis showed that thermal duration ≥10 days and atelectasis were independent risk factors for BMPs formation ( P<0.05). Conclusion:For MPP children with thermal duration≥10 days,especially with atelectasis,BMPs may have been formed,and it is recommended to perform bronchoscopy as soon as possible.

Objective:To introduce the method of using anal fistula endoscope to treat chronic sinus tract leakage at rectal anastomosis site.Methods:We used anal fistula endoscopy to treat a patient with chronic sinus tract leakage after radical resection of rectal cancer, mainly including the following 5 steps: (1) establishing a water injection circulation system through the anus; (2) scraping off purulent coating and mucosa on the surface of the sinus tract with the brush; (3) hemostasis and removal of necrotic tissue with electrocoagulation rods; (4) filling the sinus tract with bioprotein gel; (5) compressing the sinus tract with transanal drainage tube.Results:The patient is a 70 year old male with rectal cancer. After undergoing 3D laparoscopic assisted radical resection of rectal cancer via abdominal anterior resection (Dixon's procedure) and diverting ileostomy surgery for more than 3 months, leakage of the rectal anastomosis was found through colonoscopy and anal iodine water contrast imaging .The patient started eating and flowing juice 6 hours after surgery, got out of bed 24 hours after surgery, and was discharged 48 hours after the removal of the anal canal. Three months after surgery, colonoscopy and transanal iodine hydrography showed that the sinus repair was intact. The diverting ileostomy was reduced 4 months after surgery.Conclusion:Anal fistula endoscope is safe and feasible for the treatment of chronic sinus tract anastomotic leakage in selected patients.

Objective To investigate the effect and possible mechanism of microRNA-26a(miR-26a)on the syn-thesis of extracellular matrix(ECM)induced by high glucose(HG)in renal tubular epithelial cells(RTECs).Methods A model of diabetic kidney disease(DKD)was constructed by inducing RTECs with HG.MiR-26a was overexpressed in HG-induced RTECs,and RT-qPCR and Western blot were used to assess the effects of miR-26a on ECM synthesis and ferroptosis-related markers in HG-treated RTECs.Ferrostatin(Fer-1)was used to inhibit ferroptosis in the DKD model,and its impact on ECM synthesis was evaluated.RT-qPCR and Western blot were performed to measure ferroptosis-related markers,and fluorescence microscopy was used to observe the intensity of reactive oxygen species(ROS).Results Compared with the control group,the expression of miR-26a decreased in HG-treated cells,while the expression levels of ECM synthesis-related indexes fibronectin and collagen Ⅰ in-creased.After overexpressing miR-26a,the HG+miR-26a group showed a significant increase in miR-26a expres-sion and a decrease in fibronectin and collagen Ⅰ expression compared to the HG group.In terms of ferroptosis,the protein and mRNA expression of SLC7A11 and GPX4 significantly decreased,the expression of TFR-1 and AC-SL4 significantly increased,and the fluorescence intensity of ROS was significantly enhanced in the HG group com-pared with the control group.Inhibition of ferroptosis in the HG+Fer-1 group resulted in significant changes in fer-roptosis and ECM synthesis-related indicators expression levels compared to the HG group.Furthermore,re-expres-sion of miR-26a in the HG+miR-26a led to significant changes in ferroptosis-related indicators expression levels and decreased ROS fluorescence intensity compared to the HG group.Conclusions In HG-induced RTECs,miR-26a inhibits the occurrence of ferroptosis,thus reducing ECM synthesis.

Objective:To introduce the method of using anal fistula endoscope to treat chronic sinus tract leakage at rectal anastomosis site.Methods:We used anal fistula endoscopy to treat a patient with chronic sinus tract leakage after radical resection of rectal cancer, mainly including the following 5 steps: (1) establishing a water injection circulation system through the anus; (2) scraping off purulent coating and mucosa on the surface of the sinus tract with the brush; (3) hemostasis and removal of necrotic tissue with electrocoagulation rods; (4) filling the sinus tract with bioprotein gel; (5) compressing the sinus tract with transanal drainage tube.Results:The patient is a 70 year old male with rectal cancer. After undergoing 3D laparoscopic assisted radical resection of rectal cancer via abdominal anterior resection (Dixon's procedure) and diverting ileostomy surgery for more than 3 months, leakage of the rectal anastomosis was found through colonoscopy and anal iodine water contrast imaging .The patient started eating and flowing juice 6 hours after surgery, got out of bed 24 hours after surgery, and was discharged 48 hours after the removal of the anal canal. Three months after surgery, colonoscopy and transanal iodine hydrography showed that the sinus repair was intact. The diverting ileostomy was reduced 4 months after surgery.Conclusion:Anal fistula endoscope is safe and feasible for the treatment of chronic sinus tract anastomotic leakage in selected patients.

Objective:To investigate the effect of artificial dermis combined with KCI negative pressure suction device in the repair of scalp defect with skull exposure after the operation of scalp malignancy.Methods:From January 2016 to June 2018, 18 patients with scalp defect and skull exposure after scalp malignant tumor surgery were treated with artificial dermis combined with KCI negative pressure suction device.Results:Eighten patients had good wound healing after the first stage of surgery, and no infection occurred. The time for artificial dermis to complete granulation was 10-14 days, with an average of 12.1 days. After the second stage of skin grafting, the skin graft had a survival rate of 100%, with uniform color and good elasticity, without obvious scar hyperplasia or contracture. The postoperative follow-up period was 6 to 2 months, all the patients healed well without tumor recurrence, with good skin elasticity and friction-resistant, and they were satisfied.Conclusions:Artificial dermis combined with KCI negative pressure suction device is an effective and simple method to repair the scalp defect with skull exposure after tumor surgery.