Objective:To compare the differentiation efficiency of human induced pluripotent stem cell (hiPSC)-derived retinal pigment epithelium (RPE) cells using the Nuwacell ? RPE kit and nicotinamide (NIC)-based differentiation methods. Methods:Normal hiPSC lines and hiPSC lines from peripheral blood of two Bietti crystalline corneoretinal dystrophy (BCD) patients with CYP4V2 gene mutation at passages 16-18, were cultured for 4 days, and were induced into Nuwacell-RPE and NIC-RPE cells using the kit-based differentiation method and the NIC differentiation method, respectively.The percentage of pigmented cells or pigment foci areas formed at 3, 4, and 5 weeks after induction was observed and quantified under a light microscope.The cellular pigmentation level was measured by spectrophotometry.The expression of maturation markers in RPE cells was detected by real-time quantitative PCR and immunofluorescence staining.Cell polarity structures were examined by transmission electron microscopy.The induced RPE cells were co-cultured with porcine photoreceptor outer segments for 48 hours, and the number of rhodopsin (RHO)-labeled POS phagocytosed was quantified by immunofluorescence staining.Transepithelial electrical resistance was measured to evaluate the barrier function of the cells.According to the approval of the Ethics Committee of the First Affiliated Hospital of Army Medical University of China (No.[A]KY2023007), and peripheral blood from two BCD patients with CYP4V2 gene mutation was collected with written informed consent. Results:The Nuwacell method formed pigmented cells at 3 weeks, with (54.513±5.795)% pigmentation at 5 weeks, while the NIC method formed pigment spots at 4 weeks, with (26.037±8.489)% pigmentation at 5 weeks, with a statistically significant difference ( P<0.005).After 30 days, both methods produced hexagonal RPE structures, with pigmentation levels of (40.060±4.076) and (57.292±2.588)pg/cell, respectively, and the difference was statistically significant ( P<0.001).PCR and immunofluorescence staining showed that increased expression of maturation marker genes RPE65, BEST1, and CRALBP induced by the two methods.NIC-RPE cells phagocytosed more RHO-labeled outer segments than Nuwacell-RPE, showing a statistically significant difference ( t=2.920, P=0.043).After 4 weeks of culture in Transwell chambers, the transepithelial electrical resistance of NIC-RPE was higher than Nuwacell-RPE cells, demonstrating a statistically significant difference ( P<0.05).After continuous passage for 4 generations, Nuwacell-RPE partially lost its original cellular characteristics, including loss of pigmentation, irregular shape and loss of RPE marker expression, while NIC-RPE maintained its original morphologic features. Conclusions:The Nuwacell kit method has higher differentiation efficiency and RPE cell yield than the NIC-directed differentiation method, and cell maturity, phagocytosis, and barrier functions are inferior to those of the NIC-directed differentiation method.

Objective To explore the effect of visual balance training combined with digital treadmill intervention on walking func-tion in patients with ischemic stroke.Methods From July,2023 to December,2024,90 patients with ischemic stroke in Gansu Provincial Hospital were ran-domly divided into control group(n=30),treadmill group(n=30)and combined group(n=30).All groups re-ceived routine rehabilitation treatment,while the treadmill group added digital treadmill training,and the com-bined group added visual feedback balance training and digital treadmill training,for four weeks.All groups were assessed with Berg Balance Scale(BBS),Pro-Kin visual feedback balance training system,Tinetti Performance-Oriented Mobility Assessment(POMA),digital treadmill system,Timed Up and Go Test(TUGT),Fugl-Meyer Assessment-Lower Extremities(FMA-LE),Functional Ambulation Category(FAC)and modified Barthel Index(MBI)before and after intervention.Results The effects of intra-group(F>147.291,P<0.001),inter-group(F>4.919,P<0.05)and interaction(F>18.386,P<0.001)were all significant for the indicators including BBS score,length trajectory and elliptical area of eyes open or closed,POMA score,step length,hip and knee range of motion on the healthy and affected side,TUGT time,FMA-LE score,and MBI score.Post-hoc tests showed that after treatment,all the above indicators improved in each group(P<0.01),and they were the best in the combined group,followed by the treadmill group(P<0.05).After treatment,the FAC grades improved in all the groups(|Z|>1.971,P<0.05),and it was better in the combined group than in the control group(P<0.01).Conclusion Visual feedback balance training combined with digital treadmill intervention can improve balance function,walking ability and activities of daily living in patients with ischemic stroke,which is more effective than tread-mill training alone.

Background and Aims:Centrally located breast cancer(CLBC),due to its proximity to the nipple-areolar complex,has long been treated primarily with mastectomy,while the oncologic safety of breast-conserving surgery(BCS)remains controversial.This study,based on a large-scale database combined with a real-world cohort,compared the survival outcomes of BCS and mastectomy to evaluate the feasibility and oncologic safety of BCS in CLBC patients.Methods:Data of 10 325 female CLBC patients diagnosed between 2010 and 2015 were extracted from the SEER database,including 5 601 patients who underwent BCS and 4 724 who underwent mastectomy.Propensity score matching(PSM)yielded 1 951 matched pairs,and disease-specific survival(DSS)and overall survival(OS)were compared between groups.Cox regression analyses were performed to identify prognostic factors,and subgroup analyses were conducted.Additionally,an independent validation cohort from Xiangya Hospital,Central South University(2015-2016)included 221 BCS and 636 mastectomy patients,with OS and progression-free survival(PFS)assessed.Results:After PSM,baseline characteristics between groups were well balanced.Kaplan-Meier analysis demonstrated no significant differences in DSS or OS between BCS and mastectomy,and 5-,7-,and 10-year OS rates were comparable(all P＞0.05).Subgroup analyses revealed equivalent outcomes for BCS and mastectomy in patients with T1/T2 disease,different HER2 statuses,and those receiving chemotherapy,while in patients receiving radiotherapy,BCS showed significantly better DSS and OS than mastectomy(both P＜0.05).Multivariate Cox regression identified T,N,and M stage,histologic grade,molecular subtype,ER/PR status,and chemotherapy as independent prognostic factors(all P＜0.05),whereas surgical type was not(P＞0.05).The validation cohort confirmed the SEER findings,with no significant differences in OS or PFS between the two groups(both P＞0.05).Conclusions:BCS provides DSS and OS comparable to mastectomy in CLBC patients and may confer additional survival benefits when combined with radiotherapy.These findings suggest that CLBC should not be considered a contraindication to BCS,supporting BCS as a feasible and safe surgical strategy that offers valuable evidence for individualized clinical decision-making and may help improve patients' quality of life.

It is proposed that the brain Xuanfu and the glymphatic system(GS)have commonality in physiological structure and similarity in functions such as transporting brain substances,transmitting brain information and clearing metabolites.It is believed that GS may be the substantial manifestation of the brain Xuanfu,and GS dysfunction is the key pathological link of depression.Xuanfu stagnation and Shenji dysfunction are the core pathogenesis of depression.Taking"Brain Xuanfu-GS"as the starting point,the patho-logical mechanism of depression is deeply explored,providing a theoretical basis for the clinical diagnosis and treatment of depression in traditional Chinese medicine.

Objective To investigate the factors that influence ocular surface biology and visual acuity in individuals with diabetic dry eye(DDE)and analyze how these factors contribute to changes in visual quality.Methods Based on the disease duration,fasting blood glucose(FBG),and glycated hemoglobin(HbA1c)levels of patients with type 2 diabe-tes mellitus(T2DM),the DDE patients were divided into different groups.Logistic regression analysis was used to identify influencing factors related to ocular surface biology and visual quality in each group of DDE patients.Tear film stability was evaluated based on the tear film rupture time(BUT),Schirmer I test(SIt),and ocular surface disease index(OSDI).Lip-iview? Surface interferometers were used to measure tear film lipid layer thickness(LLT),meibomian gland loss rate(MGP),meibomian gland opening number(MGYLS),and meibomian gland secretion score(MGYSS).Wavefront aber-rometry was used to measure corneal wavefront aberration values at 4 mm and 6 mm pupil diameters.Ocular response ana-lyzer(ORA)was adopted to analyze corneal hysteresis(CH)and corneal resistance factor(CRF).Moreover,ELISA ex-periment to evaluate the trend of changes in inflammatory factors in tears.Results Logistic regression analysis revealed that T2DM duration,smoking history,FBG,HbA1c,total cholesterol(TC),triglycerides(TG),OSDI score,LLT,BUT,SIt,MGP,MGYLS,MGYSS,total higher-order aberrations,spherical aberration,coma aberration,trefoil aberration,tumor necrosis factor-α,interleukin-6,matrix metalloproteinase-9,receptor for advanced glycation end products,and insu-lin were all influencing factors for the risk of DDE(all P＜0.05).As the T2DM course prolonged and FBG or HbA1 c levels rose,tear film-related indicators(LLT,BUT,and SIt)and meibomian gland-related indicators(MGYLS and MGYSS)inpa-tients gradually decreased,while OSDI scores and MGP gradually increased(all P＜0.05).As the T2DM course prolonged and FBG or HbA1c levels rose,the total higher-order aberrations,spherical aberration,coma aberration,and trefoil aber-ration in DDE patients under 4 mm and 6 mm pupil diameters gradually increased;Meanwhile,best corrected visual acuity,corneal hysteresis,and corneal resistance factor gradually decreased;The contents of tumor necrosis factor-α,interleukin-6,matrix metalloproteinase-9,receptor for advanced glycation end products,and insulin in tears all gradually increased,while mucin-5AC gradually decreased(all P＜0.05).Conclusion With the prolongation of T2DM duration and the in-crease of FBG or HbA1c,the ocular surface inflammatory response in DDE patients gradually worsens,corneal biological function decreases,and visual quality deteriorates.Timely systemic and local interventions are of great significance for im-proving dry eye symptoms and visual quality in DDE patients.

Objective To investigate the relationship between the threonine and tyrosine kinase(TTK)gene and eyelid basal cell carcinoma(BCC).Methods Bioinformatics methods were used to screen the core gene(namely,TTK)associated with the occurrence and development of BCC from the Gene Expression Omnibus(GEO)database.Surgically removed eyelid BCC tissue specimens(BCC cells were divided into BBC Grade Ⅰ,Ⅱ and Ⅲ groups by tumor grade)and be-nign tumor tissue specimens(Control group)were collected from Ziyang Central Hospital for subsequent experiments.Cel-lular immunofluorescence assay(CIA)was used to detect the expression of the TTK gene in benign and malignant eyelid tumor cells.After knocking down TTK in BCC cells through transfection with lentiviruses(the cells transfected with LV-TTK-shRNA were taken as the TTK-shRNA group,and those transfected with LV-BBC-shRNA were taken as the BBC nega-tive control group),CIA was used to detect the expression of key proteins Bcl-2 and Bax in the apoptotic signaling pathway of each group of cells.Results The bioinformatics analysis showed that the TTK gene was the core gene associated with the occurrence and development of eyelid BCC.CIA detection results revealed that the fluorescence signal intensities in the tumor cytoplasm of Control,BCC Grade Ⅰ,Ⅱ,and Ⅲ groups were 1.03±0.07,1.28±0.11,1.58±0.13 and 1.92±0.17,respectively.The fluorescence signal intensity gradually increased,and the difference in fluorescence signal intensity among the four groups was statistically significant(all P＜0.05).Compared with that in the Control group(1.02±0.05),the cell fluorescence intensity was increased in the BCC negative control group(1.74±0.12)and decreased in the TTK-shRNA group(1.31±0.09)(P＜0.05).The difference in cell fluorescence intensity was significant among the Control,BCC nega-tive control and TTK-shRNA groups(all P＜0.05).The fluorescence intensity of the anti-apoptotic protein Bcl-2 was 1.04±0.12 in the Control group,2.12±0.23 in the BCC negative control group,and 1.43±0.15 in the TTK-shRNA group.The fluorescence intensity of the pro-apoptotic protein Bax was 1.02±0.08 in the Control group,0.64±0.11 in the BCC negative control group,and 1.47±0.16 in the TTK-shRNA group.After TTK knockdown,the expression level of BcL-2 in BCC cells decreased,and that of Bax increased.The fluorescence intensities of BcL-2 and Bax were significantly different among the Control,BCC negative control and TTK-shRNA groups(all P＜0.05).Conclusion The TTK gene plays a role in the regulation of eyelid BCC cell proliferation,and this effect is closely related to the PI3K-AKT-Bcl-2/Bax signaling path-way.

Objective:To compare the differentiation efficiency of human induced pluripotent stem cell (hiPSC)-derived retinal pigment epithelium (RPE) cells using the Nuwacell ? RPE kit and nicotinamide (NIC)-based differentiation methods. Methods:Normal hiPSC lines and hiPSC lines from peripheral blood of two Bietti crystalline corneoretinal dystrophy (BCD) patients with CYP4V2 gene mutation at passages 16-18, were cultured for 4 days, and were induced into Nuwacell-RPE and NIC-RPE cells using the kit-based differentiation method and the NIC differentiation method, respectively.The percentage of pigmented cells or pigment foci areas formed at 3, 4, and 5 weeks after induction was observed and quantified under a light microscope.The cellular pigmentation level was measured by spectrophotometry.The expression of maturation markers in RPE cells was detected by real-time quantitative PCR and immunofluorescence staining.Cell polarity structures were examined by transmission electron microscopy.The induced RPE cells were co-cultured with porcine photoreceptor outer segments for 48 hours, and the number of rhodopsin (RHO)-labeled POS phagocytosed was quantified by immunofluorescence staining.Transepithelial electrical resistance was measured to evaluate the barrier function of the cells.According to the approval of the Ethics Committee of the First Affiliated Hospital of Army Medical University of China (No.[A]KY2023007), and peripheral blood from two BCD patients with CYP4V2 gene mutation was collected with written informed consent. Results:The Nuwacell method formed pigmented cells at 3 weeks, with (54.513±5.795)% pigmentation at 5 weeks, while the NIC method formed pigment spots at 4 weeks, with (26.037±8.489)% pigmentation at 5 weeks, with a statistically significant difference ( P<0.005).After 30 days, both methods produced hexagonal RPE structures, with pigmentation levels of (40.060±4.076) and (57.292±2.588)pg/cell, respectively, and the difference was statistically significant ( P<0.001).PCR and immunofluorescence staining showed that increased expression of maturation marker genes RPE65, BEST1, and CRALBP induced by the two methods.NIC-RPE cells phagocytosed more RHO-labeled outer segments than Nuwacell-RPE, showing a statistically significant difference ( t=2.920, P=0.043).After 4 weeks of culture in Transwell chambers, the transepithelial electrical resistance of NIC-RPE was higher than Nuwacell-RPE cells, demonstrating a statistically significant difference ( P<0.05).After continuous passage for 4 generations, Nuwacell-RPE partially lost its original cellular characteristics, including loss of pigmentation, irregular shape and loss of RPE marker expression, while NIC-RPE maintained its original morphologic features. Conclusions:The Nuwacell kit method has higher differentiation efficiency and RPE cell yield than the NIC-directed differentiation method, and cell maturity, phagocytosis, and barrier functions are inferior to those of the NIC-directed differentiation method.

Oral squamous cell carcinoma(OSCC)is a malignant lesion originating from the oral mucosal squamous epithelium,account-ing for over 80％of oral and maxillofacial malignancies.Key etiological factors include tobacco,alcohol abuse,and betel quid chewing.In China,its incidence has shown an overall upward trend,posing a significant threat to public health.OSCC exhibits high local invasive-ness,making early diagnosis critical for improving prognosis.Its clinical management requires close multidisciplinary collaboration among oral and maxillofacial surgery,head and neck surgery,radiation oncology,medical oncology,reconstructive surgery,radiology,patholo-gy,and nutritional support teams.Given the increasing disease burden of OSCC and rapid development of multidisciplinary collaborative models,an expert panel has formulated this integrated management consensus based on evidence-based medicine and extensive deliber-ation.Centered on the'Prevention-Screening-Diagnosis-Treatment-Rehabilitation'framework,the consensus provides comprehensive guidance for the entire disease course of OSCC patients,aiming to standardize clinical practice.

Objective:To screen and extract relevant evidence on the management of malnutrition in pediatric cancer patients and provide a best evidence summary.Methods:A systematic search was conducted across multiple websites and databases, including UpToDate, BMJ Best Practice, WHO website, Guidelines International Network, National Institute for Health and Care Excellence, National Guideline Clearinghouse, PubMed, Web of Science Core Collection, Embase, Cochrane Library, China National Knowledge Infrastructure, China Biology Medicine disc, Wanfang Data and others, for evidence on nutritional management of malnutrition in pediatric cancer patients. The search included literature from inception to August 31, 2023. Literature was selected following strict inclusion and exclusion criteria by researchers trained in evidence-based nursing courses. The quality of the selected literature was evaluated, and evidence was extracted and summarized.Results:A total of 11 articles were included, comprising two clinical decision papers, three guidelines, one evidence summary, two systematic reviews, and three expert consensus documents. The evidence was summarized into 24 evidence across five main areas: multidisciplinary team approach, nutritional risk screening and assessment, nutrient intake, dietary and nutritional education, and enteral and parenteral nutrition support.Conclusions:This study provides a best evidence summary for the nutritional management of malnutrition in pediatric cancer patients, offering evidence-based support for clinical practice among healthcare professionals.

Objective:To test the difficulty, discrimination, and reliability of the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire.Methods:Two researchers independently translated the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire and cross-checked it to form a Chinese version of the questionnaire. The Chinese version of the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire consists of 24 items, with correct answers scoring one point and incorrect answers scoring zero points, with a total score of 24 points. Convenience sampling was used to select ICU nurses from 14 GradeⅢ Class A hospitals in five provinces/autonomous regions and two municipalities in China for the survey between April and July 2023. The difficulty index, discrimination index, and Cronbach's α coefficient of the questionnaire were analyzed.Results:A total of 1 121 questionnaires were distributed, with 1 020 valid responses, yielding a valid response rate of 90.99%. The mean score of the 1 020 ICU nurses on the Chinese version of the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire was (16.10±5.58) , with a minimum score of 4.00 and a maximum score of 24.00. The Cronbach's α coefficient of the Chinese version of the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire was 0.91. The questionnaire's overall difficulty and discrimination indexes were 0.67 and 0.59, respectively.Conclusions:The Chinese version of the ICU Nurse Pressure Injury Prevention and Care Knowledge Questionnaire has appropriate difficulty, moderate discrimination, and strong reliability, making it a valuable tool for assessing ICU nurses' knowledge of pressure injury-related topics.