Hepatitis B virus （HBV） exclusively infects liver parenchymal cells and forms covalently closed circular DNA （cccDNA） within their nuclei. HBV cccDNA serves as the essential template for viral gene transcription， the sole source of progeny virus production， and the key driver of viral antigen expression， and it is the molecular basis for the persistence of HBV infection. Therefore， elimination and/or functional silencing of cccDNA is the key to eradicate chronic HBV infection. This article discusses the critical scientific issues that need to be solved during elimination of the harm of HBV infection from the perspectives of the synthesis， transcription， and clearance of cccDNA， as well as the impact of nonparenchymal cells on cccDNA， in order to provide a reference for eradicating HBV infection in the future.

ObjectiveTo investigate the effect and mechanism of long intergenic non-coding RNA02086 （LINC02086） overexpression mediated macrophage polarization on the proliferation， migration and invasion of gastric cancer cells. MethodsThe expression levels of LINC02086 in the human gastric epithelial cell line GES-1 and human gastric cancer cell lines HCG-27， NCI-N87， and AGS were determined by qRT-PCR. Human acute monocytic leukemia cells （THP-1） were induced to differentiate into M0 macrophages using phorbol 12-myristate 13-acetate （PMA）. HGC-27 cells were infected with either LINC02086 overexpression lentivirus （OE-LINC02086） or its negative control lentivirus （Vector）， and the culture supernatants were collected as conditioned medium （CM1）. M0 macrophages were co-cultured with the infected HGC-27 cells， and the resulting supernatants were designated as conditioned medium 2 （CM2）. M0 macrophages were treated with CM1 alone or in combination with Wnt/β-catenin pathway inhibitor IWR-1， forming the Vector+CM1， OE-LINC02086+CM1， and OE-LINC02086+CM1+IWR-1 groups， respectively. Flow cytometry was used to detect mannose receptor C-type 1 （CD206） expression， and qRT-PCR was employed to measure mRNA levels of interleukin-10 （IL⁃10）， transforming growth factor-β （TGF⁃β）， vascular endothelial growth factor （VEGF）， and chemokine ligand 22 （CCL22）. Western blot was performed to evaluate protein expression of CD206， VEGF， and key components of the Wnt/β-catenin pathway—Wnt family member 3a （Wnt3a）， glycogen synthase kinase-3β （GSK-3β）， and β-catenin. HGC-27 cells were treated with CM2 alone or combined with IWR-1， establishing the Vector+CM2， OE-LINC02086+CM2， and OE-LINC02086+CM2+IWR-1 groups. CCK-8 assay was used to evaluate cell proliferation， and Transwell assays were conducted to assess migration and invasion capabilities. ResultsCompared with GES-1 cells， the expression levels of LINC02086 were upregulated in HCG-27， NCI-N87， and AGS cells （P < 0.05）， with the smallest increase observed in HCG-27 cells. Compared with Vector+CM1 group， the level of CD206 and the expression levels of IL⁃10， TGF⁃β， VEGF and CCL22 mRNA in macrophages stimulated by OE-LINC02086+CM1 increased （P<0.05）. Meanwhile， the expression levels of Wnt3a and β-catenin proteins in cells increased （P<0.05）， and the expression level of GSK-3β protein decreased （P<0.05）. However， co-treatment with IWR-1 markedly reversed the promoting effects of LINC02086 overexpression on the expression of M2 polarization markers， including CD206， IL⁃10， and TGF⁃β mRNA， in macrophages （P<0.05）， as well as its activation of the Wnt/β-catenin signaling pathway （P<0.05）. Compared with Vector+CM2 group， HGC-27 cells infected with OE-LINC02086+CM2 had increased proliferation activity and increased number of migration and invasion cells （P<0.05）. However， the combined intervention of IWR-1 significantly reversed the promotion of LINC02086 overexpression on the proliferation， migration and invasion of HGC-27 cells （P<0.05）. ConclusionLINC02086 overexpression promotes the proliferation， migration and invasion of gastric cancer cells by activating Wnt/β-catenin pathway to mediate M2 polarization of macrophages.

Both type 2 diabetes mellitus (T2DM) and metabolic associated fatty liver disease (MAFLD) fall within the spectrum of metabolic diseases, and they exhibit a bi-directional causal relationship and robust reciprocal association. Their shared pathological cornerstone is insulin resistance (IR), which involves the interplay of mitochondrial dysfunction and chronic inflammation, forming a cascading pathological process of “IR-mitochondrial dysfunction-inflammation”. This largely explains the notable upward trend in T2DM-MAFLD co-occurrence observed over recent years. Exercise intervention, as a safe and effective non-pharmacological approach, can improve the pathological progression of these patients at multiple levels. Following the logical framework of “pathogenesis-efficacy comparison-molecular mechanisms-clinical translation”, this article systematically compares the efficacy and molecular mechanisms of moderate-intensity continuous training (MICT), resistance exercise, high-intensity interval training (HIIT), and combined training. MICT reduces intrahepatic triglycerides by promoting lipolysis and improving cardiorespiratory fitness; resistance exercise increases muscle mass and basal metabolic rate, offering unique advantages in preserving muscle while reducing fat and improving insulin sensitivity; HIIT is a time-efficient exercise modality that enhances patients’ cardiorespiratory fitness and insulin sensitivity by alternating brief periods of vigorous exertion with recovery periods, with a prominent short-term triglyceride-lowering effect; combined training produces synergistic effects, comprehensively improving glucolipid metabolism and showing the best long-term adherence. Mechanistically, exercise exerts its beneficial effects through three common pathways: (1) AMPK-mediated lipid oxidation and mitochondrial biogenesis; (2) IRS/PI3K/Akt-mediated insulin signaling sensitization; and (3) Nrf2/ARE anti-oxidation and TGF-β/Smads anti-fibrosis regulation. Different exercise modalities activate these pathways with distinct emphases: MICT most directly and persistently activates the AMPK pathway; resistance exercise uniquely improves IRS/PI3K/Akt signaling through muscle mass gain; HIIT induces the highest AMPK activation intensity and triggers unique lactate-mediated signaling regulation; combined training integrates the above multiple mechanistic advantages. For clinical translation, multidisciplinary team collaboration is essential to ensure safety and adherence. Individualized prescriptions should be formulated according to the FITT-VP principle and patient phenotypes—frequency of 3-5 sessions/week of aerobic exercise combined with 2-3 sessions/week of resistance exercise; intensity of moderate-intensity (40%-60% heart rate reserve (HRR))aerobic exercise and 60%-80% of one-repetition maximum (1-RM) for resistance exercise; time of at least 150 min/week of moderate-intensity aerobic exercise, 30-60 min per session; type of combined training as the preferred modality; total volume of≥500-1 000 MET-min/week; and progression adjusted every 4-6 weeks—with real-time adjustments supported by wearable devices, ultimately forming a closed-loop management system from initial assessment to long-term follow-up. Notably, current studies have limitations such as small sample sizes and short intervention periods. Future research should focus on long-term follow-up, multi-omics biomarkers, and combined exercise-drug strategies. In conclusion, the systematic integration of structured, individualized, and sustainable exercise interventions into the multidisciplinary management pathway for patients with T2DM complicated by MAFLD is an urgent need in current clinical practice.

Objective To investigate the distribution characteristics of treatment time for total marrow irradiation (TMI) or total marrow and lymphoid irradiation (TMLI) based on helical tomotherapy, establish a time parameter framework for the application of TMI/TMLI techniques, and provide a basis for optimizing clinical treatment efficiency, predicting patient tolerance, and streamlining workflow. Methods A retrospective analysis was conducted on 57 patients who received TMI/TMLI using helical tomotherapy between November 2024 and July 2025. Patients were stratified by height ( ≤ 130 cm group vs. > 130 cm group). Megavoltage computed tomography scanning time, beam-on time, and total treatment time were recorded. The relationship between height and treatment time was analyzed using Spearman correlation analysis. An independent samples t-test was used to compare treatment time between TMI and TMLI modes. Results In the ≤ 130 cm group, the mean megavoltage computed tomography scanning time, beam-on time, and total treatment time per fraction were (9.67 ± 1.47), (39.96 ± 9.08), and (49.63 ± 10.16) minutes, respectively. In the > 130 cm group, the corresponding times were (14.52 ± 1.17), (60.45 ± 11.19), and (74.97 ± 11.82) minutes, respectively. Treatment time was positively correlated with height (r = 0.756, P < 0.001). Among patients taller than 130 cm, there was no statistically significant difference in treatment time between the TMI and TMLI (P > 0.05). Conclusion Although helical tomotherapy requires a relatively long treatment time, its technical characteristics are well-suited for the extensive and complex target volumes involved in TMI/TMLI. Future technological upgrades and standardized stratification hold promise for enhancing both efficiency and precision, thereby expanding clinical applicability.

Objective To investigate the expression of kinesin family member 15(KIF15), epidermal growth factor receptor(EGFR), and human epidermal growth factor receptor 2(HER2) in gallbladder cancer(GBC) and their clinical and pathological significance. Methods Immunohistochemical staining was employed to detect the expression of KIF15, EGFR, and HER2 proteins in GBC tissue microarrays. The correlation between protein expression levels and various clinical and pathological characteristics of GBC patients was analyzed. Results The positive expression rates of KIF15, EGFR, and HER2 proteins in GBC tissues were 71.6%, 62.2%, and 51.4% respectively, compared to 16.7%, 0, and 0 in para-carcinoma tissues (all P=0.000). KIF15 expression was correlated with cancer differentiation grade (P=0.006), while EGFR expression was associated with lymph node metastasis(P=0.026) and the number of metastatic lymph nodes(P=0.012). HER2 expression was correlated with cancer differentiation grade(P=0.018), tumor size(P=0.005), and T stage(P=0.032). GBC patients with co-positive expression of KIF15 and HER2 had significantly lower overall survival compared to those with negative expression. Positive HER2 expression, lymph node metastasis, and the number of metastatic lymph nodes were identified as independent risk factors for overall survival in GBC patients. Conclusions Positive expression of KIF15, EGFR, and HER2 in GBC tissues are correlated with various clinical indicators and poorer prognosis. These proteins may serve as potential factors for predicting prognosis and evaluating therapeutic efficacy in GBC.

Objective:To investigate the efficacy and safety of venetoclax and azacitidine combined with GHA (human granulocyte colony stimulating factor, homoharringtonine and low-dose cytarabine) priming regimen in treatment of patients with relapsed/refractory acute myeloid leukemia.Methods:A retrospective case series study was conducted. Twenty-three patients with relapsed/refractory acute myeloid leukemia (non-acute promyelocytic leukemia) who received treatment with the combination of venetoclax and azacitidine with GHA priming regimen at the Second Affiliated Hospital of Xi'an Jiaotong University from October 2020 to July 2024 were selected, and the treatment efficacy, minimal residual disease (MRD)-negative rate in patients with comprehensive complete remission (cCR) (including complete remission, complete remission with partial hematologic recovery and complete remission with incomplete hematologic recovery) and the adverse reactions were analyzed; patients were followed-up, and their overall survival (OS) was analyzed by using Kaplan-Meier method.Results:The median age of the 23 patients was 60 years (range: 21-79 years), including 10 males and 13 females. The cCR rate for 1 course of treatment was 52.2% (12/23), with 4 cases of MRD negative among cCR patients; 5 cases received 2 courses of treatment, with 3 cases achieving cCR, of which 2 cases were MRD negative; 2 cases received 3 courses of treatment, with 1 case achieving complete remission with incomplete hematologic recovery. Six patients underwent allogeneic hematopoietic stem cell transplantation. The patients were followed up until July 31, 2024, and the median follow-up period was 5.3 months (range: 1.1-41.7 months). Ten cases survived, 12 cases died, 1 case was lost to follow-up, and the median OS time of 23 patients was 7.9 months. The 6-month OS rate was 60.2% (95% CI: 42.7%-84.8%), and the 12-month OS rate was 44.6% (95% CI: 26.8%-74.3%). Common adverse reactions during treatment included infection [69.6% (16/23)], nausea [56.5% (13/23)], febrile neutropenia [52.2% (12/23)], bleeding [52.2% (12/23)], vomiting [34.8% (8/23)], and pneumonia [34.8% (8/23)]. Conclusions:The combination of vinaclotide and azacitidine with GHA priming regimen has certain efficacy and good safety in the treatment of relapsed/refractory acute myeloid leukemia.

Objective:To explore the application effect of early enteral nutrition combined with optimized blood glucose management in patients with stress hyperglycemia after cardiac surgery.Methods:A study was conducted to construct an early enteral nutrition and blood glucose optimization management plan for patients with stress hyperglycemia after cardiac surgery.A prospective analysis was conducted from May 2022 to May 2023 in the Cardiothoracic Surgery Intensive Care Unit(CICU)of a tertiary hospital in Shanghai.200 patients with stress hyperglycemia after cardiac surgery were divided into control group(n=100)and observation group(n=100)by random number table method.The control group received routine enteral nutrition and blood glucose management plan,while the observation group received early enteral nutrition combined with blood glucose optimization management plan.Nutrition related outcome indicators,blood glucose management quality indicators,and patient prognosis indicators were compared between two groups of patients.Results:After implementing the early enteral nutrition combined with optimized blood glucose management plan,the nutritional indicators(serum prealbumin,hemoglobin,and 7-day calorie compliance rate)of the observation group were higher than those of the control group,and the differences were statistically significant(P<0.05);The maximum blood glucose fluctuation amplitude,insulin use time,and incidence of hypoglycemia in the observation group were significantly lower than those in the control group(P<0.05),and the time within the glucose target range was significantly higher than that in the control group,with statistical significance(P<0.05);The mechanical ventilation time,ICU stay time,and total hospital stay in the observation group were significantly lower than those in the control group,and the differences were statistically significant(P<0.05).Conclusions:Early enteral nutrition combined with optimized blood glucose management can effectively improve the nutritional status of patients with stress hyperglycemia after cardiac surgery,reduce blood glucose fluctuations,improve blood glucose management quality,and promote patient outcomes and prognosis.

Objective:To evaluate the impact of different levels of enteral caloric support on clinical outcomes in patients with Stevens-Johnson Syndrome(SJS)/Toxic Epidermal Necrolysis(TEN).Methods:A retrospective analysis was conducted in 125 patients with SJS/TEN admitted to the Third People's Hospital of Hangzhou from January 2013 to December 2023.Patients were categorized into three groups based on their daily enteral caloric intake:low-calorie group(<25 kcal/kg,n=75),conventional calorie group(25～30 kcal/kg,n=27),and high-calorie group(>30 kcal/kg,n=23).The inflammatory markers,nutritional indicators,skin lesion healing time,length of hospital stay,complications rates,and mortality were compared among the groups.Results:At discharge,the white blood cell count[WBC,(6.08±0.60)109/L]and C-reactive protein[CRP,(19.04±7.09)mg/L]in the conventional calorie group,as well as the high-calorie group[WBC(6.34±0.71)109/L,CRP(27.78±8.94)mg/L],were significantly lower than those in the low-calorie group[WBC(8.31±0.41)109/L、CRP(62.07±7.41)mg/L](P<0.05).Meanwhile,the albumin[Alb(34.92±0.84)g/L]and prealbumin[PA,(243.48±12.38)mg/L]levels in the conventional calorie group,as well as the high-calorie group[Alb(35.07±0.96)g/L;PA(250.30±12.32)mg/L],were significantly higher than those in the low-calorie group[Alb(27.31±0.38)g/L;PA(146.31±6.01)mg/L](P<0.05).The skin lesion healing time(11.00±3.84)days,length of hospital stay(14.93±4.68)days,in the conventional calorie group were significantly lower than those in the low-calorie group[(skin lesion healing time(15.52±4.61)days,length of hospital stay(19.92±6.17)days,P<0.05];the skin healing time(10.6±2.96)d and length of hospital stay(14.07±4.05)d in the high-calorie group were significantly lower than those in the low-calorie group(P<0.05).There was no significant difference in infection rates(8.70%vs 8.00%)and gastrointestinal intolerance rates(8.70%vs 10.67%)between the two groups(P>0.05).Conclusion:Enteral caloric support of 25～30 kcal/kg/day appears to be optimal for patients with SJS/TEN,effectively meeting energy demands,improving nutritional and inflammatory markers,and promoting better clinical outcomes with fewer complications.

Objective To investigate the diversity and composition of gut fungi microbiota in patients with polycystic kidney disease(PKD)and its correlation with clinical indicators.Methods A total of 44 PKD patients,44 patients with non-polycystic chronic kidney disease(NPCKD)and 22 healthy controls(HC)admitted to our hospital from February 2023 to February 2024 were recruited.ITS1 DNA sequencing was applied to analyze the gut fungal composition.Bioinformatics analysis was used to compare the diversity and structural differences of fungi among the 3 groups.Pearson correlation analysis was performed to analyze the relationship between gut fungi and clinical indicators.Results There were no significant differences in baseline characteristics(gender,age,body mass index,etc.)among the 3 groups,but statistical differences were seen in terms of serum indicators(such as serum creatinine,blood urea nitrogen,uric acid,estimated glomerular filtration rate,etc.)(P<0.01).Alpha diversity analysis showed no significant difference was seen between the PKD and HC groups,but the PKD group had significant differences to the NPCKD group(P<0.01).Beta diversity analysis revealed significant differences among the 3 groups and in pairwise comparisons(P=0.001).Fungi composition analysis found that the abundance of Candida was significantly higher in the PKD group than the other 2 groups(P<0.01),while the abundances of Aspergillus and Cladosporium were significantly lower in the PKD group than the HC group(P<0.05).Linear discriminant analysis(LEFSe)indicated that Candida was significantly enriched,while Aspergillus and Cladosporium were significantly reduced in the PKD group.Correlation analysis revealed that the abundance of Cladosporium was negatively correlated with cyst diameter and immunoglobulin light chain Kappa/Lambda ratio in the PKD group(P<0.05),while the abundance of Candida was positively correlated with liver/kidney cyst diameter(P<0.01).Conclusion PKD patients exhibit characteristic changes in gut fungi diversity and composition.The abundances of Cladosporium and Candida are closely associated with clinical indicators of PKD patients.

Objective To analyze the diversity and composition of gut microbiota in individuals with circadian rhythm disruption and their correlation with cognition.Methods Night shift workers and regular shift workers were subjected from our hospital during August 2022 and October 2024.The participants with circadian rhythm disorders were assigned into an experimental group(n=24),and those with normal circadian rhythms were into a control group(n=24).Their height,weight,age,gender,body mass index(BMI)and fresh fecal samples were collected,and Montreal Cognitive Assessment(MoCA)and Mini-Mental State Examination(MMSE)were used to evaluate their mental status.Metagenomics,Alpha and Beta diversity analyses,Linear Discriminant Analysis Effect Size(LEfSe),and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis were employed to investigate the diversity and function characteristics of gut microbiota in the participants.Results There were no statistical differences between the 2 groups in baseline data,such as height,weight,gender,age,and BMI(P>0.05).Alpha diversity analysis indicated that no statistical differences were observed in the ACE,Chao1,Shannon,or Simpson indices between the 2 groups,while beta diversity analysis revealed significant differences(P<0.01),suggesting different structure of gut microbiota between them.In the experimental group,the abundance of Faecalibacterium prausnitzii and Agathobacter rectalis was decreased,while that of Escherichia coli and Phocaeicola vulgaratus was increased,with significant differences when compared with the control group(P<0.05).Additionally,KEGG functional analysis showed that the experimental group had obviously higher expression levels in Th17 cell differentiation and the IL-17 signaling pathway than the control group(P<0.05).Agathobacter rectalis and Faecalibacterium prausnitzii were positively correlated with MoCA score and MMSE score(P<0.05,P<0.01).Agathobacter rectalis was negatively correlated with the IL-17 signaling pathway and Th17 cell differentiation.Conclusion Individuals with circadian rhythm disorders have significant changes in the structure and function of gut microbiota when compared to those with normal circadian rhythms.Agathobacter rectalis may be involved in the regulation of the IL-17 signaling pathway and differentiation of Th17 cells,thereby possibly impacting the increases of cognitive score related to circadian rhythm disorders.