Objective To investigate the role and potential mechanism of secreted phosphoprotein 1 (SPP1)⁺ macrophages in the formation of chronic renal allograft fibrosis. Methods The expression features of SPP1⁺ macrophages in renal allografts of chronic allograft dysfunction (CAD) patients were analyzed based on single-cell transcriptome data of renal tissues from patients with CAD. Transcription factor VIPER analysis and DoRothEA transcription factor activity analysis were performed on the single-cell transcriptome data. Renal tissue samples were collected from kidney transplant recipients, including the CAD group (n=5) and the non-renal allograft fibrosis group (CTL group, n=5). A mouse model of chronic allograft rejection was established and divided into the allogeneic kidney transplantation group (CAD group, n=3) and the syngeneic kidney transplantation group (SYN group, n=3). Hematoxylin-eosin staining was used to detect renal tissue injury in mice, and Masson staining was used to detect renal tissue fibrosis. Immunofluorescence staining was performed to detect SPP1 expression in renal tissues of transplant recipients and mouse renal allografts. Bone marrow-derived macrophages (BMDMs) were extracted from mice and subjected to hypoxia stimulation. The expression of hypoxia-inducible factor (HIF)-1α and SPP1 was detected by Western blot, and SPP1 expression was detected by flow cytometry. BMDMs were transfected with HIF-1α overexpression plasmid and HIF-1α small interfering RNA (siRNA) followed by hypoxia intervention, and the expression of HIF-1α and SPP1 was detected by Western blot. Mouse aortic endothelial cells (MAECs) were co-cultured with the supernatant of BMDMs, and the expression of endothelial-mesenchymal transition (EndMT)-related markers was detected by Western blot and immunofluorescence. Results Single-cell transcriptome analysis showed that the proportion of SPP1⁺ macrophages in renal allograft tissues was significantly higher in the CAD group than in the CTL group (P<0.05). The renal injury score and the percentage of interstitial fibrotic area in the CAD group were significantly higher than those in the SYN group (both P<0.05). Immunofluorescence staining showed that the proportion of SPP1⁺ macrophages was increased in the CAD group compared with the CTL group, and also increased in the CAD group compared with the SYN group (both P<0.05). VIPER analysis and DoRothEA transcription factor activity analysis revealed activation of the hypoxia pathway and upregulated expression of transcription factors such as HIF-1α in SPP1⁺ macrophages. SPP1 expression was elevated in BMDMs under hypoxic conditions. Knockdown of HIF-1α inhibited hypoxia-induced SPP1 protein expression, whereas overexpression of HIF-1α upregulated SPP1 protein levels. After co-culture of hypoxia-induced BMDMs with MAECs, the expression levels of EndMT-related markers were increased. Conclusions SPP1⁺ macrophages differentiated under hypoxia are significantly infiltrated in the formation of chronic renal allograft fibrosis, and may promote renal allograft fibrosis by inducing EndMT in renal vascular endothelial cells.

HBV DNA integration （iDNA） is the core barrier that must be overcome to achieve functional cure for chronic hepatitis B （CHB） and to prevent the occurrence of hepatocellular carcinoma （HCC）. During reverse transcription， 5%　—　10% of viral genomes are converted into double-stranded linear DNA that is randomly inserted into host chromosomes， generating stable iDNA and continuously producing HBsAg， thereby driving B- and T-cell immune exhaustion and locking the host in an immune-tolerant state. The process of iDNA runs throughout the entire natural history of HBV infection， and the viral enhancers it carries can promote clonal hyperplasia of indeterminate potential， accumulate pre-neoplastic mutations， and ultimately lead to HCC. Although long-term nucleos（t）ide analog or interferon therapy can suppress viral replication and reduce the formation of new integrations， existing therapies still fail to eliminate existing iDNA. Therefore， there is an urgent need for innovative strategies that can precisely target integration breakpoints， epigenetically silence iDNA， or eradicate integrated clones， so as to significantly increase the functional cure rate of CHB and fundamentally reduce the risk of HCC.

Objective To investigate the association of serum total testosterone(TT),free testosterone(FT),sex hormone-binding globulin(SHBG),and estradiol(E_2)with all-cause mortality rate(AMR)in males and to evaluate their predictive value. Methods Based on data from the National Health and Nutrition Examination Survey(NHANES)cycles(1999—2004 and 2013—2016),the relationships between different sex hormones and AMR was analyzed with Cox proportional hazards regression models. The dose-response relationships between different sex hormones and male AMR were assessed with restricted cubic splines(RCS). The predictive performances of different sex hormones for male AMR were compared with receiver operating characteristic(ROC)curves and the area under the curve(AUC). The survival differences across varying sex hormone levels were evaluated with Kaplan-Meier survival curves and log-rank tests. Results This study involved 3632 adult male participants with a median follow-up of 6.5 years,during which 320 AMR occurred(8.8%). Cox univariate analysis indicated that TT,FT and SHBG were risk factors for AMR(P＜0.001). Cox multivariate analysis revealed that FT and SHBG were independent risk factors for AMR(FT:HR=0.90,95%CI:0.86-0.94; SHBG:HR=1.01,95%CI:1.01-1.02). RCS revealed a U-shaped nonlinear relationship between TT,FT and AMR(P＜0.001); SHBG showed a positive dose-response relationship with AMR(P＜0.001). The AUCs of FT predicting the 5-year,10-year,and 15-year AMR were 0.78(95%CI:0.73-0.82),0.80(95%CI:0.76-0.84),and 0.84(95%CI: 0.80-0.88),respectively,significantly outperforming other indicators. Subgroup analysis revealed that FT was negatively correlated with male AMR,but this association was modulated by age,race,smoking status and diabetes status(P_(interaction)＜0.05). Kaplan-Meier survival analysis showed that different levels of TT and FT(Q1-Q4)were significantly positively correlated with survival rate(P＜0.001),while different levels of SHBG were significantly negatively correlated with survival rate,with patients in the Q4 group having the highest risk of death(P＜0.001). Conclusion Based on US NHANES,studies have shown that the FT and SHBG are independent risk factors for male AMR,and FT demonstrates greater predictive value for male AMR than TT.

Chronic hepatitis B （CHB） is a significant public health issue worldwide. The persistent presence of covalently closed circular DNA of the hepatitis B virus （HBV） and the integration of the viral genome are the main obstacles limiting the achievement of virological cure in CHB. Functional cure has now become an important goal in antiviral therapy for CHB， and its accurate determination and efficacy prediction rely on a comprehensive assessment of multidimensional diagnostic markers. This article systematically reviews the research advances in diagnostic markers associated with functional cure in CHB， focusing on the clinical value of serological and virological indicators （HBsAg quantification and HBV DNA）， genomic， transcriptomic， proteomic， and metabolomic markers， and indicators associated with host innate immunity and adaptive immunity. Furthermore， it discusses future research directions such as the establishment of unified detection standards and the construction of predictive models combining multiple markers， in order to promote the precise assessment and individualized management of functional cure in CHB.

Purpose To investigate interhemispheric homotopic functional connectivity in patients with neuropsychological systemic lupus erythematosus(NPSLE)during resting state and its relationship with clinical indicators and neuropsychological scales.Materials and Methods This prospective study enrolled 35 patients with NPSLE and 31 healthy controls(control group)from Taizhou People's Hospital Affiliated to Nanjing Medical University(June 2020 to March 2023).All participants underwent resting state functional MRI and completed neuropsychological assessments including mini-mental state examination,Montreal cognitive assessment,hospital anxiety and depression scale,fatigue scale for motor and cognitive functions,along with laboratory tests(C3,C4,IgA,IgM,IgG).Image preprocessing and voxel-mirrored homotopic connectivity(VMHC)calculations were performed using DPABI V7.0 on Matlab R2013b.Between-group differences in VMHC values were compared,and correlations between VMHC values in significant regions and neuropsychological/clinical data were analyzed.Results The NPSLE group demonstrated significantly lower mini-mental state examination and Montreal cognitive assessment scores compared with those in control group(t=-6.297,-7.001,both P=0.001).Patients with NPSLE exhibited significantly decreased VMHC values in bilateral parahippocampal gyri,precentral gyri,middle frontal gyri,and medial/paracingulate gyri compared with those in control group(family-wise error corrected,voxel-level P<0.001,cluster-level P<0.05).In the NPSLE group,VMHC values in precentral gyri showed positive correlation with IgA levels(r=0.351,P=0.039),while VMHC values in medial/paracingulate gyri positively correlated with IgA(r=0.345,P=0.043)and negatively with C4(r=-0.368,P=0.030).Conclusion Patients with NPSLE demonstrate abnormal interhemispheric homotopic functional connectivity,and the correlation between imaging metrics and clinical data in differential brain regions may facilitate early diagnosis of NPSLE while providing novel insights into the neuropathological mechanisms of cerebral injury.

Objective To investigate the effects of miR-362-3p on the proliferation,migration and invasion of esophageal cancer EC9706 cells and its molecular mechanism.Methods The cancerous tissues and adjacent tissues of 30 patients with esophageal cancer from January 2018 to January 2020 were selected,and human normal esophageal epithelial cells HET-1 A and esophageal cancer cells EC9706,TE10,KYSE-140,KYSE-150 were cultured in vitro.Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression levels of miR-362-3p and dual specificity phosphatase 10(DUSP10)mRNA in tissues and cells.EC9706 cells were divided into NC group(normally cultured),miR-NC group(transfected miR-362-3p mimic NC)and miR-362-3p group(transfected miR-362-3p mimic),si-NC group(transfected with DUSP10 siRNA NC),si-DUSP10 group(transfected with DUSP10 siRNA),miR-362-3p+pcDNA group(co-transfected with miR-362-3p mimic and pcDNA3.1-DUSP10 NC)and miR-362-3p+pcDNA-DUSP10 group(co-transfected with miR-362-3p mimic and pcDNA3.1-DUSP10).The expression levels of miR-362-3p and DUSP10 mRNA in each group were detected by RT-qPCR.Cell proliferation activity was detected by MTT assay.Cell migration and invasion were detected by Transwell assay.The expressions of DUSP10,CyclinD1,p21,matrix metalloproteinase(MMP)-2 and MMP-9 were detected by Western blot.Dual luciferase reporter gene assay was used to detect the targeting relationship between miR-362-3p and DUSP10.Results The expression of miR-362-3p was high and DUSP10 was low in esophageal cancer tissues and cells.Overexpression of miR-362-3p or knockdown of DUSP10 expression significantly reduced the activity,migration and invasion number and the expression of CyclinD1,MMP-2 and MMP-9,and significantly increased the expression of p21 in EC9706 cells(P＜0.05).miR-362-3p targeted the expression of DUSP10(P＜0.05).And up-regulation of DUSP10 expression partially reversed the effects of overexpression of miR-362-3p on the proliferation,migration and invasion of EC9706 cells(P＜0.05).Conclusion miR-362-3p can inhibit the proliferation,migration and invasion of EC9706 cells by down-regulating the expression of DUSP10.

Objective To investigate the effects of miR-362-3p on the proliferation,migration and invasion of esophageal cancer EC9706 cells and its molecular mechanism.Methods The cancerous tissues and adjacent tissues of 30 patients with esophageal cancer from January 2018 to January 2020 were selected,and human normal esophageal epithelial cells HET-1 A and esophageal cancer cells EC9706,TE10,KYSE-140,KYSE-150 were cultured in vitro.Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression levels of miR-362-3p and dual specificity phosphatase 10(DUSP10)mRNA in tissues and cells.EC9706 cells were divided into NC group(normally cultured),miR-NC group(transfected miR-362-3p mimic NC)and miR-362-3p group(transfected miR-362-3p mimic),si-NC group(transfected with DUSP10 siRNA NC),si-DUSP10 group(transfected with DUSP10 siRNA),miR-362-3p+pcDNA group(co-transfected with miR-362-3p mimic and pcDNA3.1-DUSP10 NC)and miR-362-3p+pcDNA-DUSP10 group(co-transfected with miR-362-3p mimic and pcDNA3.1-DUSP10).The expression levels of miR-362-3p and DUSP10 mRNA in each group were detected by RT-qPCR.Cell proliferation activity was detected by MTT assay.Cell migration and invasion were detected by Transwell assay.The expressions of DUSP10,CyclinD1,p21,matrix metalloproteinase(MMP)-2 and MMP-9 were detected by Western blot.Dual luciferase reporter gene assay was used to detect the targeting relationship between miR-362-3p and DUSP10.Results The expression of miR-362-3p was high and DUSP10 was low in esophageal cancer tissues and cells.Overexpression of miR-362-3p or knockdown of DUSP10 expression significantly reduced the activity,migration and invasion number and the expression of CyclinD1,MMP-2 and MMP-9,and significantly increased the expression of p21 in EC9706 cells(P＜0.05).miR-362-3p targeted the expression of DUSP10(P＜0.05).And up-regulation of DUSP10 expression partially reversed the effects of overexpression of miR-362-3p on the proliferation,migration and invasion of EC9706 cells(P＜0.05).Conclusion miR-362-3p can inhibit the proliferation,migration and invasion of EC9706 cells by down-regulating the expression of DUSP10.

This study aims to investigate the mechanisms underlying the effects of the combined ac-tion of high-fat diet-induced obesity and the aflatoxin B1(AFB1)on hepatic oxidative stress and inflammatory responses.Thirty-six rats of similar weight and 4 weeks old were randomly divided into 4 groups,with 9 rats in each group:the blank control group(basal diet),the AFB1 group(0.4 mg/kg AFB1+basal diet),the HFD group(high-fat diet),and the HFD+AFB1 group(high-fat diet+0.4 mg/kg AFB1).Histological changes and lipid deposition were observed via hematox-ylin-eosin(HE)staining and Oil Red O staining.Levels of oxidative stress and inflammation-relat-ed factors were measured using commercial assay kits.The relative protein expression levels of factors involved in the Nrf2-Keap1 signaling pathway were assessed by Western blot analysis.The HE staining results showed that in the AFB1 group,the liver cells exhibited widespread watery de-generation,with shrunk and ruptured nuclei,inflammatory cells infiltration,and increased fibrosis.In the HFD group,liver cell fatty degeneration was observed,with cytoplasmic lipid droplet infil-tration.In the portal area,liver fibrosis was seen,with liver cell necrosis and inflammatory cell in-filtration in the fibrotic area,accompanied by lipofuscous granules.When HFD was applied to the AFB1 group,the abnormal state of liver interstitial and interstitial spaces was further aggravated,and a large number of lipid droplets appeared.The Oil Red O staining results showed that there were large numbers of dark red lipid droplets in the liver tissue of the HFD group,which were fused in strands.In the AFB1 group,lipid droplets could also be observed in the liver tissue of rats,but the number and degree were significantly less than those in the HFD group.The number and degree of red lipid droplets in the liver tissue of rats in the HFD+AFB1 group were higher than those in the AFB1 group and the HFD group.HFD exacerbated AFB1-induced oxidative stress by elevating ROS,MDA levels,and decreasing the expression of antioxidant stress factors such as CAT,SOD,Nrf2,HO-1,NQO1,and GCLC.Furthermore,the combined effect of HFD and AFB1 further significantly increased the levels of pro-inflammatory cytokines IL-2,IL-6,TNF-α,and IL-1β in the body.In summary,HFD treatment significantly exacerbated liver oxidative stress and in-flammatory responses in rats exposed to AFB1 through the Nrf2-keap1 signaling pathway.

Objective:To explore the effects of holmium laser enucleation of the prostate with early complete transection of the urethral mucosa at the tip of the prostate on urinary control function and sexual function in patients with benign prostatic hyperplasia (BPH).Methods:Eighty patients with BPH who underwent holmium laser enucleation of the prostate in the Affiliated Hospital of Yan′an University from January 2019 to January 2023 were collected as the research subjects. The patients were divided into the observation group and the control group by the random number table method, with 40 cases in each group. The observation group underwent early holmium laser enucleation of the prostate with early complete transection of the urethral mucosa at the tip of the prostate, while the control group underwent conventional holmium laser prostatectomy. The general conditions, urinary control function and sexual function of the two groups of patients after the operation were compared. The adverse ejaculation conditions 6 months after the operation were recorded.Results:There was no statistically significant difference in age and prostate volume between the two groups of patients (all P>0.05). The operation time, intraoperative blood loss, postoperative indwelling urinary catheter time and postoperative hospital stay in the observation group were significantly less than those in the control group (all P<0.05). The International Prostate Symptom Scale (IPSS) score, Quality of Life (QOL) score, the maximum flow rate (Qmax), and post void residual (PVR) in the bladder of the two groups of patients 6 months after the operation were compared with those before the operation, and the differences were statistically significant (all P<0.05), while there were no statistically significant differences between the groups (all P>0.05). There were no statistically significant differences in the International Index of Erectile Function (IIEF-5) scores and Erection Hardness Grading Scale (EHGS) grades of the two groups of patients 6 months after surgery compared with those before surgery (all P>0.05), and there were also no statistically significant differences between the groups (all P>0.05). There was no statistically significant difference in the ejaculation function score and ejaculation distress score 6 months after the operation in the observation group compared with those before the operation (all P>0.05), while in the control group, the ejaculation function score 6 months after the operation was lower than that before the operation, and the ejaculation distress score was higher than that before the operation (all P<0.05). The ejaculation function score and ejaculation distress score of the observation group 6 months after the operation were significantly better than those of the control group (all P<0.05). The incidences of retrograde ejaculation and reduced semen volume 6 months after the operation in the observation group were both lower than those in the control group (all P<0.05). There was no statistically significant difference in the incidence of rapid ejaculation, ejaculation pain, hematospermia, etc. between the two groups of patients 6 months after the operation (all P>0.05). Conclusions:In holmium laser enucleation of the prostate, early complete transection of the urethral mucosa at the tip of the prostate has an improving effect on urinary control function and sexual function in patients with BPH, and increases the confidence in postoperative life and satisfaction with orgasm of BPH patients.

Objective:To study the risk factors of thyroid nodules in patients with type 2 diabetes.Methods:Data of patients with type 2 diabetes with normal thyroid function admitted to the Department of Endocrinology of Heze Municipal Hospital from January to June 2024 were collected. Binary logistic regression was used to analyze the influencing factors of thyroid nodules in patients with type 2 diabetes, and the receiver operating characteristic curve (ROC curve) was used to evaluate the diagnostic efficacy of each influencing factor.Results:Among 162 patients with type 2 diabetes, 96 had thyroid nodules, accounting for 59.3%. The incidence of thyroid nodules in women was significantly higher than that in men (χ 2 = 4.56, P = 0.034). Multivariate logistic regression analysis showed that age (≥50 years old), overweight and obesity [body mass index (BMI)≥24.0 kg/m 2], high glycated hemoglobin (≥10%), and high total cholesterol ( > 6.5 mmol/L) were independent risk factors for thyroid nodules in patients with type 2 diabetes ( OR = 1.83, 1.67, 1.08, 3.65, P < 0.05), and men was an independent protective factor ( OR = 0.63, P = 0.039). The ROC curve results showed that total cholesterol and total cholesterol combined with glycated hemoglobin could distinguish patients with thyroid nodules from those without thyroid nodules, with AUC = 0.64 and 0.68, respectively, and the differences were statistically significant ( P < 0.05). Conclusions:The incidence of thyroid nodules in patients with type 2 diabetes is relatively high. Age, overweight and obesity, high glycated hemoglobin, and high total cholesterol are independent risk factors for thyroid nodules in patients with type 2 diabetes, and total cholesterol has the ability to distinguish patients with thyroid nodules.