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 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.

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 detect the expression of N-acetyltransferase 10(NAT10)and polyadenylate bind-ing protein cytoplasmic 1(PABPC1)in non muscle invasive bladder cancer(NMIBC),and analyze the correla-tion between them and epithelial mesenchymal transition(EMT)and prognosis.Methods A total of 122 pa-tients with NMIBC treated in the hospital from May 2019 to May 2021 were selected.Immunohistochemistry was used to detect the expression of NAT10 and PABPC1 proteins in NMIBC tissues.Real time fluorescence quantitative polymerase chain reaction(qPCR)was used to detect the expression of NAT10,PABPC1 mRNA,and EMT markers in NMIBC tissues.Pearson correlation analysis was conducted on the correlation between EMT indicators[Snail,N-cadherin(N-cad),vimentin(Vim)mRNA].Cox regression analysis was conducted on the relationship between NAT10,PABPC1 and prognosis of NMIBC.Results Compared with adjacent tis-sues,the expression of NAT10 mRNA,PABPC1 mRNA,Snail mRNA,N-cad mRNA,and Vim mRNA in NMIBC cancer tissues was higher,and the difference was statistically significant(P＜0.001).The expression of NAT10 mRNA,PABPC1 mRNA in NMIBC cancer tissues was positively correlated with Snail mRNA,N-cad mRNA,and Vim mRNA(r=0.678,0.702,0.711,0.754,0.788,0.663,P＜0.001).The positive rates of NAT10 and PABPC1 in NMIBC cancer tissues were 59.02％(72/122)and 60.66％(74/122),respectively,while those in adjacent tissues were 6.56％(8/122)and 4.92％(6/122),respectively(x2=76.176,85.995,P＜0.001).The positive rates of NAT10 and PABPC1 in NMIBC cancer tissues were higher than those in ad-jacent tissues,and the difference was statistically significant(x2=76.176,85.995,P＜0.001).The positivity rates of NAT10 and PABPC1 in cancer tissues of stage T1,high-grade NMIBC patients were higher than those in cancer tissues of Ta/Ti,low-grade patients,and the differences were statistically significant(P＜0.05).The 3-year overall progression free survival rates of NMIBC patients in the NAT10 positive and negative groups were 48.61％(35/72)and 80.00％(40/50),respectively,with a statistically significant difference(Log rank x2=13.780,P=0.000).The 3-year overall progression free survival rates of PABPC1 positive and negative patients were 47.30％(35/74)and 83.33％(40/48),respectively,with a statistically significant difference(Log rank x2=11.830,P=0.001).T1 stage,high-grade,NAT10 positive,and PABPC1 positive were risk fac-tors affecting the prognosis of NMIBC.Conclusion The expression of NAT10 and PABPC1 in NMIBC cancer tissue is significantly upregulated and positively correlated with EMT markers,which is correlated with poor prognosis of NMIBC.

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.

Objective:To explore the mechanisms of action of Huanglian Jiedu decoction combined with Xijiao Dihuang decoction (HLJDT-XJDH) in regulating fibroblasts in the treatment of psoriasis. Methods:A mouse model of psoriasis was established by topical application of imiquimod 5% cream on the shaved back; HLJDT-XJDH at different doses of 7.7 and 30.6 g/kg was administered via gavage for intervention, and methotrexate (2 mg/kg) served as a positive control; after 7 days, the severity of skin lesions was assessed using the psoriasis area and severity index (PASI), while histopathological changes of skin tissues were evaluated using hematoxylin-eosin (HE) staining and Baker scoring. For in vitro experiments, fibroblasts were divided into a control group, a model group, a low-dose (5% drug-containing serum) intervention group, and a high-dose (20% drug-containing serum) intervention group; cells in the control group were cultured with 20% normal rat serum for 24 hours; in the model group, cells cultured with 20% normal rat serum were stimulated with 5 ng/ml tumor necrosis factor (TNF) -α and 50 ng/ml interleukin (IL) -17A for 24 hours to mimic fibroblasts during the occurrence of psoriasis; cells in the low- and high-dose intervention groups received the same stimulation as the model group, and were cultured for 24 hours with 5% and 20% HLJDT-XJDH-containing serum, respectively, but not with the 20% normal rat serum. After the above treatment, these cells were co-cultured with keratinocytes (HaCaT cells) using a Transwell system. In addition, on the basis of the control group, fibroblasts were divided into the model group, 20% drug-containing serum intervention group, and 20% drug-containing serum intervention + OE-SFRP2 group; TNF-α and IL-17A were used to stimulate the cells to simulate the psoriatic state; the treatment in the 20% drug-containing serum intervention group was carried out as previously described; in the 20% drug-containing serum intervention + OE-SFRP2 group, cells were transfected with the vector for 48 hours to establish an overexpression model, followed by culture with 20% drug-containing serum for 24 hours, without co-culture with HaCaT cells.. Cell counting kit-8 (CCK-8) assay was performed to assess cell viability, flow cytometry to measure apoptosis rates, enzyme-linked immunosorbent assay (ELISA) to detect levels of inflammatory cytokines (TNF-α, IL-1β, IL-6) as well as chemokine ligand (CXCL) 1 and CXCL12 in mouse serum or cell culture supernatant, qPCR to determine the mRNA expression of inflammatory cytokines, chemokines, cell cycle- and proliferation-related factors, as well as SFRP2 in mouse skin tissues or cells, and Western blot analysis to determine the protein expression of SFRP2, Wnt3a, and β-catenin in fibroblasts. One-way analysis of variance was employed for intergroup comparisons, and post-hoc analysis was conducted using Tukey's test. Results:In vivo mouse experiments showed that compared with the normal control group, the model group exhibited typical psoriatic characteristics in skin morphology, including significant inflammatory infiltration in skin tissues and marked epidermal thickening; compared with the normal control group, the serum levels of TNF-α (531.16 ± 28.27 pg/ml vs. 239.58 ± 10.39 pg/ml), IL-1β (111.40 ± 5.16 pg/ml vs. 80.35 ± 3.87 pg/ml), and IL-6 (109.17 ± 4.84 pg/ml vs. 71.73 ± 2.04 pg/ml) significantly increased in the model group, along with their mRNA expression levels in mouse skin tissues (all P < 0.001) ; compared with the model group, the treatment group showed alleviated psoriatic manifestations, and significant reductions in the levels of inflammatory factors TNF-α (low-dose, high-dose, and positive control groups: 420.80 ± 29.30 pg/ml, 322.33 ± 9.40 pg/ml, 322.97 ± 12.16 pg/ml, respectively), IL-1β (98.69 ± 4.49 pg/ml, 89.02 ± 1.56 pg/ml, 88.88 ± 2.08 pg/ml, respectively), and IL-6 (94.07 ± 3.76 pg/ml, 80.54 ± 3.30 pg/ml, 83.21 ± 3.18 pg/ml, respectively), as well as in their mRNA expression levels (all P < 0.001). In in vitro fibroblast experiments, compared with the control group, the model group exhibited a significant elevation in the supernatant levels of IL-1β (126.42 ± 3.56 pg/ml vs. 34.81 ± 0.44 pg/ml), IL-6 (459.44 ± 9.35 pg/ml vs. 115.51 ± 7.26 pg/ml), CXCL1 (2 434.88 ± 127.63 pg/ml vs. 762.85 ± 30.60 pg/ml) and CXCL12 (3 542.14 ± 35.86 pg/ml vs. 2 095.86 ± 45.12 pg/ml), the expression levels of their mRNAs (all P < 0.001), as well as the protein expression levels of SFRP2, Wnt3a, and β-catenin; after intervention with HLJDT-XJDH-containing serum, all the above indices significantly decreased (all P < 0.001). However, when 20% drug-containing serum intervention was administered simultaneously, the expression of inflammatory factors and chemokines in fibroblasts was significantly higher in the SFRP2 overexpression group than in the non-overexpression group (all P < 0.01). When fibroblasts were co-cultured with HaCaT cells, the model group showed significantly increased cell viability but a decreased apoptosis rate of HaCaT cells compared with the control group, while the low- and high-dose intervention groups showed significantly decreased cell viability but increased apoptosis rates of HaCaT cells compared with the model group (all P < 0.05) . Conclusion:HLJDT-XJDH may exert therapeutic effects in psoriasis by downregulating the SFRP2/Wnt/β-catenin signaling pathway, thereby inhibiting fibroblast activation and inflammatory process, which subsequently suppresses the proliferation of keratinocytes and the activation of inflammatory cells.

Objective To describe the clinical features of patients with primary sclerosing cholangitis(PSC)in China based on a nationwide multicenter patient cohort,and to investigate the risk factors for prognosis.Methods A retrospective cohort study was conducted among the patients with a confirmed diagnosis of PSC based on the electronic medical record system of seven grade A tertiary hospitals across the country,and related data were extracted.The Mann-Whitney U test was used for comparison of continuous data between groups,and the chi-square test was used for comparison of categorical data between groups.The Kaplan-Meier method was used to estimate liver transplant-free survival,and the log-rank test was used for comparison of survival rate between PSC patients with different features.The Cox regression model was used to identify independent risk factors for the prognosis of PSC patients and the interactions between key factors.Results A total of 107 patients were enrolled,among whom 55.6%(55/99)had large-duct PSC and 29.0%(31/107)had comorbidity with inflammatory bowel disease(IBD).The positivity rate of anti-neutrophil cytoplasmic antibody(ANCA)was 32.9%(24/73),and 50.0%(40/80)of the patients had an increase in IgG/IgM.The median symptom-to-diagnosis interval was 1 year(<1-4.0),and 38.3%(41/107)of the patients had progressed to decompensated cirrhosis at the time of diagnosis.The median liver transplant-free survival time was 114 months(95%confidence interval[CI]:62-166),with a 5-year survival rate of 65.7%.The multivariate analysis showed that an increase in total bile acid(TBA)(hazard ratio[HR]=1.006,95%CI:1.002-1.010,P=0.001)and a prolonged symptom-to-diagnosis interval(HR=1.252,95%CI:1.059-1.480,P=0.009)were independent risk factors for prognosis.The interaction analysis showed that compared with the female patients with TBA<50 μmol/L,both male and female patients with TBA≥50 μmol/L had a significant increase in the risk of liver transplantation or death(male:HR=16.563,95%CI:2.103-130.449,P<0.001;female:HR=17.009,95%CI:2.113-136.934,P<0.001),and compared with the patients with an age of<45 years and a TBA level of<50 μmol/L,the patients with an age of≥45 years and a TBA level of≥50 μmol/L had a significant increase in the risk of liver transplantation or death(HR=10.729,95%CI:1.325-86.859,P=0.026).Compared with the female patients with an symptom-to-diagnosis interval of≤2 years,the male patients with a symptom-to-diagnosis interval of>2 years had an increased risk of liver transplantation or death(HR=4.825,95%CI:1.725-13.644,P=0.003),and compared with the patients with an age of<45 years and a symptom-to-diagnosis interval of≤2 years,the patients with an age of<45 years and a symptom-to-diagnosis interval of>2 years had an increased risk of liver transplantation or death(HR=4.983,95%CI:1.366-18.173,P=0.015).Conclusion Compared with the reports from Western countries,large-duct PSC is also the main type of PSC in China,but with a relatively low proportion,and there is also a relatively low proportion of patients with IBD or positive ANCA.An increase in TBA and a prolonged symptom-to-diagnosis interval are independent risk factors for prognosis,with significant interactions with age and sex.This suggests that early screening and intervention should be enhanced to improve prognosis.

Objective To investigate the effect of lung lobe surface projection localization combined with lung segment drainage sputum technique on airway clearance in patients with aspiration pneumonia,providing a reference for clinical nursing practice.Methods A convenience sampling method was used to select 62 patients with perioperative aspiration pneumonia secondary to brain diseases admitted to a tertiary A hospital in Beijing from May 2022 to October 2024 as the research subjects.A total of 31 patients admitted from August 2023 to October 2024 were assigned to a control group,and 31 patients admitted from May 2022 to July 2023 were assigned to an experimental group.The experimental group received lung lobe surface projection positioning combined with lung segment drainage sputum technique on top of conventional back percussion sputum clearance technique,while the control group received conventional back percussion sputum clearance technique.After the intervention,the differences in oxygenation state,inflammatory test indicators,treatment efficiency,vital signs and frequency of suction coughing between 2 groups were compared.Results After the intervention,the experimental group showed better oxygenation index,frequency of suction coughing,white blood cell count,percentage of neutrophils,procalcitonin levels,interleukin-6 levels,and treatment efficiency compared to those in the control group(P＜0.05).There was no statistically significant difference in heart rate,respiratory rate,and C-reactive protein levels between 2 groups(P＞0.05).Conclusion Lung lobe surface projection positioning combined can effectively promote the patients with aspiration pneumonia of loosening of sputum in the lungs,improve airway clearance efficiency,enhance the patient's pulmonary ventilation and gas exchange capacity,improve oxygenation levels,and reduce systemic inflammatory levels.

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.