Background/Aims: Natural killer (NK) cell function is generally considered dampened in chronic hepatitis B virus (HBV) infection; however, the NK cell pool exhibits phenotypic and functional heterogeneity, and the antibody--mediated effect of NK cells remains less characterized. This study evaluated the dynamic changes in antibody-mediated NK cell responses and the involvement of distinct NK subsets across disease stages and during antiviral treatment. Methods: A T-cell receptor-like antibody specific for the HBV core 18–27 peptide (cTCRL-Ab) was used to determine the antibody-mediated effect of NK cells, and an array of NK cell surface markers were analyzed in cross-sectional and longitudinal cohorts of patients with chronic HBV infection. Single-cell RNA sequencing (scRNA-seq) was performed to identify the heterogeneity of NK subsets. Results: The cTCRL-Ab enabled the detection of NK cell cytolytic activity and IFNγ production. Notably, cTCRL-Ab-mediated NK cell responses were compromised in chronically HBV-infected patients, particularly in those receiving pegylated interferon-α (Peg-IFNα), which was associated with the downregulation of CD16 expression. Correspondingly, Peg-IFNα inhibited cTCRL-Ab-mediated NK cell function by reducing CD16 expression in vitro. scRNA-seq revealed that CD16 downregulation occurred mainly within a dysfunctional CD16hi NK subset exhibiting exhaustion properties. In contrast, an activated CD16hiNK subpopulation (CX3CR1⁺KLRC2–CD16hi) with high cytotoxicity was enriched in patients who experienced favorable treatment responses. Furthermore, the intrahepatic CX3CR1+KLRC2–CD16hi subset tended to exhibit functional restoration in HBsAg-loss individuals. Conclusions Our data contribute to the understanding of antibody-mediated responses of NK cells in chronic HBV infection, and highlight a previously unappreciated functional CX3CR1+KLRC2–CD16hiNK subset as a potential therapeutic target.

ObjectiveTo investigate whether intestinal deficiency of lanosterol synthase （LSS）， a key enzyme in cholesterol synthesis， influences the progression of metabolic dysfunction-associated steatotic liver disease （MASLD） by regulating intestinal cholesterol absorption and immune response. MethodsLSS heterozygous knockout （LSS^+/-） mice and wild-type （WT） controls were generated using CRISPR/Cas9 technology and fed either a high-fat diet （HFD） or regular chow （CHOW）. The model was validated by genotyping. Hepatic steatosis was assessed by HE and oil red O staining. Immunohistochemistry was used to detect the localization and expression of NPC1L1 and CD36 proteins in the intestine. Western blot analysis was performed to measure JNK phosphorylation and TLR4 protein levels in intestinal tissues. Real-time quantitative polymerase chain reaction （qPCR） was employed to examine the mRNA expression of TLR4 and IL-6. ResultsLSS^+/- mice were successfully validated by genotyping and reduced intestinal LSS protein expression. HE and oil red O staining of liver sections showed that， compared with WT mice fed a CHOW diet， WT mice fed a HFD exhibited a marked increase in hepatic lipid vacuoles. In contrast， compared with HFD-fed WT mice， HFD-fed LSS^+/- mice displayed significantly attenuated hepatic lipid deposition and reduced serum ALT levels （P<0.05）. Immunohistochemical analysis revealed that， compared with WT mice， the expression of the cholesterol absorption protein NPC1L1 in the intestinal villi of LSS^+/- mice was downregulated under both CHOW and HFD conditions （P_HFD<0.001）. Conversely， the expression of the fatty acid transporter CD36 was upregulated in the intestines of LSS^+/- mice （P_CHOW<0.05， P_HFD<0.01）. Western blot analysis demonstrated that， compared with WT mice， TLR4 protein expression in the intestines of LSS^+/- mice significantly increased under both CHOW and HFD conditions （both P<0.05）. JNK phosphorylation level was significantly elevated in LSS^+/- mice under CHOW condition （both P<0.05）. Under HFD condition， total JNK protein expression increased， but its phosphorylation level showed no significant change. qPCR analysis showed that， compared with WT mice， the mRNA levels of TLR4 （P_CHOW<0.01， P_HFD<0.000 1） and IL-6 （P_CHOW<0.001， P_HFD<0.01） were significantly upregulated in the intestines of LSS^+/-mice. ConclusionLSS deficiency counteracts hepatic lipid deposition by orchestrating a synergistic reprogramming involving restricted intestinal cholesterol absorption， enhanced fatty acid utilization， and activation of immune pathways， suggesting intestinal LSS as a potential therapeutic target of MASLD.

To evaluate the application value of the Peyton four-step teaching method in the standardized training of intestinal ultrasound and compare it with traditional teaching methods, so as to provide an optimized approach for clinical ultrasound training.

ObjectiveTo investigate the effect of hypoxia on pemetrexed （PE）-mediated apoptosis in non-small cell lung cancer cells. MethodsA549 cells were cultured under 21% O₂ （normoxia） or 1% O₂ （hypoxia）， and treated with 8 μmol/L PE for 48 h. The cells were divided into normoxic control group， normoxic vehicle group， normoxic drug-treated group， hypoxic control group， hypoxic vehicle group and hypoxic drug-treated group. Trypan blue staining was used to observe cell death after PE intervention under normoxia and hypoxia. Flow cytometry was adopted to detect cell apoptosis rate. Wound healing assay was used to assess cell migration. Western blot was performed to detect the protein levels of MRP-1， Cleaved Caspase-3， E-Cadherin and N-Cadherin. ResultsTrypan blue staining results showed that the cell death rate was reduced in PE-treated cells under hypoxia compared with the normoxic PE group （P<0.001）. Flow cytometry results indicated that PE induced cell apoptosis under both normoxic and hypoxic conditions.However， the apoptosis rate of the normoxic PE group was significantly higher than that of the hypoxic PE group （P<0.000 1）. Wound healing assay showed that the migration distance of A549 cells decreased after PE treatment for 24 h and 48 h under normoxia and hypoxia when compared with the control group （P<0.05）， while no significant difference was found between the normoxic PE group and the hypoxic PE group. Western blot results revealed that MRP-1 expression was markedly higher in hypoxic PE-treated cells than in normoxic PE-treated cells （P<0.000 1）. Compared with the control group， Cleaved Caspase-3 expression was upregulated in A549 cells following PE intervention under normoxia （P<0.05）， whereas no obvious alteration in Cleaved Caspase-3 expression was detected in hypoxic PE-treated cells. Under both normoxic and hypoxic conditions， PE treatment downregulated the expression of E-Cadherin （P<0.000 1，P<0.001） and N-Cadherin （P<0.05，P<0.000 1）. In addition， the expression levels of E-Cadherin and N-Cadherin in the hypoxic PE group were significantly lower than those in the normoxic PE group （all P<0.000 1）. ConclusionHypoxia significantly attenuates the pro-apoptotic effect of PE on A549 cells and blocks Cleaved Caspase-3 activation. This mechanism may be related to hypoxia-induced MRP-1 overexpression and reduced drug accumulation. Interfering with MRP-1 or restoring the activity of the apoptosis pathway may serve as a novel strategy to overcome hypoxia-induced drug resistance.

Objective: To analyze the status of outdoor activities on weekends among children and adolescents of different educational stages in Shanghai and their impact on myopia, so as to provide a basis for formulating more specific prevention and control protocol of myopia. Methods: From September to October 2022, a stratified cluster random sampling method was employed to select 84 schools (27 kindergartens, 21 primary schools, 15 junior high schools and 21 high schools) across Shanghai, enrolling a total of 28 654 children and adolescents aged 4 to 18 for the study. Ophthalmic examinations were conducted to ascertain the prevalence of myopia among children and adolescents. Additionally, a questionnaire survey was administered to collect data on outdoor activity duration and associated factors. Multivariate Logistic regression analysis was utilized to investigate the associated factors of outdoor activity levels on weekends. Results: The overall myopia detection rate among children and adolescents was 58.4%, with a higher rate observed in girls (59.2%) compared to boys (57.6%). The myopia detection rates for children and adolescents with an average daily outdoor activity duration of ≥2 h and <2 h on weekends were 54.6% and 68.8%, and the differences were statistically significant ( χ 2=8.12,460.89, P <0.01). Multivariable Logistic regression analysis revealed that girls ( OR =0.80), those with a myopic parent ( OR =0.68), schools from urban districts ( OR =0.72), higher education stages (primary school: OR =0.65, junior high school: OR =0.24, high school: OR =0.14) and spending≥2 h/d on homework during weekends ( OR =0.57) among children and adolescents were less likely to engage in outdoor activities for ≥2 h on weekends ( P <0.01). After incorporating gender, parental myopia status, educational stage, school location, average daily duration on weekends for spending on homework, electronic product usage and outdoor activities as dependent variables in a multivariate Logistic regression analysis, the results showed that children and adolescents with an average outdoor activity duration for ≥2 h on weekends had a lower risk of myopia ( OR =0.86, P < 0.01). Conclusions The level of outdoor activity among children and adolescents on weekends needs to be improved. Outdoor activities on weekends is an associated factor for myopia among children and adolescents. Particularly, girls, those with myopic parents, schools from urban districts, and spending long hours on homework during weekends among children and adolescents require increased attention.

Objective:To construct a device-related pressure injury prevention program for orthopedic patients based on evidence-based and SSKINS (S: skin, S: surface, K: keep moving, I: incontinence, N: nutrition, S: self-care) clinical management model, and to provide reference for clinical nursing practice.Methods:In June 2023, a research team was set up to retrieve the literature related to device-related pressure injury in orthopedic patients, and the evidence was extracted and screened with reference to the SSKINS clinical management model, and the first draft of the protocol was formulated based on clinical practice. The final plan was determined through two rounds of expert consultations from October to December 2023.Results:The response rate of the two rounds of expert correspondence was 100%(20/20). The expert authority coefficients were 0.897 and 0.907, the overall Kendall coordination coefficients of the two rounds were 0.116 and 0.136 respectively, and the Kendall coordination coefficients of all levels were 0.104-0.213 (all P<0.001). The final protocol contains 6 first-level items (assessment and examination, the use and adjustment of device, prophylactic dressings, skin protection, nutritional support, health education) and 31 second-level items. Conclusions:The proposed orthopedic device-related pressure injury prevention program is scientific and practical, and can provide a reference for the prevention and management of clinical medical staff.

Objective To understand the current situation of thirst in patients with cirrhosis and analyze its influencing factors,in order to improve medical staff's attention to thirst symptoms in patients with cirrhosis and provide theoretical basis for clinical intervention.Methods A total of 220 patients with cirrhosis who were hospitalized in the infection department of a tertiary A general hospital in Wuhan from March to June 2024 were selected by convenience sampling method.General data questionnaire,Numerical Score Scale and Thirst Distress Scale were used to investigate the factors affecting thirst sensation in patients with cirrhosis.Results A total of 202 valid questionnaires were collected,and the effective questionnaire recovery rate was 91.82％.The results showed that the incidence of thirst in patients with cirrhosis was 59.41％;the score of thirst was 3.00(3.00,6.00)points,and the mean score of thirst was 3.09.Among them,43.07％of the patients with cirrhosis were in the moderate to severe level of thirst.The score of Thirst Distress Scale is 6.00(6.00,17.00)points,and the average score of thirst distress was 11.62 points,which was in the medium level.47.03％of patients with cirrhosis indicated thirst distress,and 23.76％of patients with cirrhosis which was in the moderate and severe level of distress.The results of multiple linear regression analysis showed that the degree of ascites,the use of diuretics,the stage of disease,and the degree of thirst distress were the factors influencing the degree of thirst in patients with cirrhosis(P＜0.05).Gender,marital status,degree of ascites,use of diuretic drugs,disease stage and degree of thirst were the factors influencing degree of thirst distress in patients with cirrhosis(P＜0.05).Conclusion The incidence and severity of thirst in patients with cirrhosis are relatively high,and are affected by many factors.Medical staff should pay more attention to the management of thirst symptoms in patients with cirrhosis,and formulate targeted nursing measures or nursing programs according to the related influencing factors of thirst,so as to improve the comfort level of patients and improve the disease experience of patients.

Objective:To observe the early clinical efficacy of 3D-printed modular supporting prosthesis for reconstruction of lacunar bone defect caused by giant cell tumor of the distal femur.Methods:From May 2018 to July 2023, a total of 9 patients with giant cell tumor of the distal femur were treated with 3D-printed modular supporting prosthesis to reconstruct lacunar bone defects in the Department of Orthopedics, West China Hospital, Sichuan University. There were 4 males and 5 females, aged 30.8±6.1 years (range, 24-44 years), 5 cases on the left side, 4 cases on the right side, 2 cases of Campanacci grade I, 7 cases of Campanacci grade II. The anteroposterior and lateral X-ray films and T-SMART tomosynthesis imaging of the knee joint were taken to observe the bone graft healing and osseointegration after operation. Musculoskeletal Tumor Society (MSTS)-93 was used to evaluate knee function, and visual analogue scale (VAS) was used to evaluate knee pain.Results:All patients were successfully operated and followed up for an average of 30.8±7.5 months (range, 18-42 months). The operation time was 124.2±23.6 min, and the intraoperative blood loss was 105.6±17.4 ml. All autografts showed bony union at the graft-host junction, and the healing time was 3.3±0.4 months (range, 3.0-4.0 months). At 6 months after surgery, T-SMART tomosynthesis imaging showed that the gap between the prosthesis-bone interface was less than 1 mm in all patients. At the last follow-up, the thickness of residual subchondral bone was 5.7±1.3 mm, which was greater than that before operation 2.2±0.8 mm, and the difference was statistically significant ( t=10.823, P<0.001). At the last follow-up, the score of MSTS-93 was 26.7±2.4, which was higher than that before operation 18.8±3.7, and the difference was statistically significant ( t=5.367, P<0.001). At the last follow-up, the range of motion of the knee joint was 122.8°±9.1°, which was higher than that before operation 108.3°±6.1°, and the difference was statistically significant ( t=3.970, P<0.001). All patients were able to walk normally, go up and down stairs and other daily activities, and 7 patients were able to complete squats. At the last follow-up, there was no local tumor recurrence, distant metastasis, death, joint infection, pain (VAS score was 0), delayed wound healing, joint degeneration, prosthesis loosening or articular surface collapse. Conclusion:Reconstruction of lacunar bone defect caused by giant cell tumor of distal femur with 3D-printed modular supporting prosthesis can effectively improve knee joint function and osseointegration, and the short-term clinical results are satisfactory.

Objective To understand the current situation of thirst in patients with cirrhosis and analyze its influencing factors,in order to improve medical staff's attention to thirst symptoms in patients with cirrhosis and provide theoretical basis for clinical intervention.Methods A total of 220 patients with cirrhosis who were hospitalized in the infection department of a tertiary A general hospital in Wuhan from March to June 2024 were selected by convenience sampling method.General data questionnaire,Numerical Score Scale and Thirst Distress Scale were used to investigate the factors affecting thirst sensation in patients with cirrhosis.Results A total of 202 valid questionnaires were collected,and the effective questionnaire recovery rate was 91.82％.The results showed that the incidence of thirst in patients with cirrhosis was 59.41％;the score of thirst was 3.00(3.00,6.00)points,and the mean score of thirst was 3.09.Among them,43.07％of the patients with cirrhosis were in the moderate to severe level of thirst.The score of Thirst Distress Scale is 6.00(6.00,17.00)points,and the average score of thirst distress was 11.62 points,which was in the medium level.47.03％of patients with cirrhosis indicated thirst distress,and 23.76％of patients with cirrhosis which was in the moderate and severe level of distress.The results of multiple linear regression analysis showed that the degree of ascites,the use of diuretics,the stage of disease,and the degree of thirst distress were the factors influencing the degree of thirst in patients with cirrhosis(P＜0.05).Gender,marital status,degree of ascites,use of diuretic drugs,disease stage and degree of thirst were the factors influencing degree of thirst distress in patients with cirrhosis(P＜0.05).Conclusion The incidence and severity of thirst in patients with cirrhosis are relatively high,and are affected by many factors.Medical staff should pay more attention to the management of thirst symptoms in patients with cirrhosis,and formulate targeted nursing measures or nursing programs according to the related influencing factors of thirst,so as to improve the comfort level of patients and improve the disease experience of patients.

Objective:To construct a device-related pressure injury prevention program for orthopedic patients based on evidence-based and SSKINS (S: skin, S: surface, K: keep moving, I: incontinence, N: nutrition, S: self-care) clinical management model, and to provide reference for clinical nursing practice.Methods:In June 2023, a research team was set up to retrieve the literature related to device-related pressure injury in orthopedic patients, and the evidence was extracted and screened with reference to the SSKINS clinical management model, and the first draft of the protocol was formulated based on clinical practice. The final plan was determined through two rounds of expert consultations from October to December 2023.Results:The response rate of the two rounds of expert correspondence was 100%(20/20). The expert authority coefficients were 0.897 and 0.907, the overall Kendall coordination coefficients of the two rounds were 0.116 and 0.136 respectively, and the Kendall coordination coefficients of all levels were 0.104-0.213 (all P<0.001). The final protocol contains 6 first-level items (assessment and examination, the use and adjustment of device, prophylactic dressings, skin protection, nutritional support, health education) and 31 second-level items. Conclusions:The proposed orthopedic device-related pressure injury prevention program is scientific and practical, and can provide a reference for the prevention and management of clinical medical staff.