Immunotherapy has emerged as a revolutionary approach to treat immune-related diseases. Dendritic cells (DCs) play a pivotal role in orchestrating immune responses, making them an attractive target for immunotherapeutic interventions. Modulation of gene expression in DCs using genome editing techniques, such as the CRISPR-Cas system, is important for regulating DC functions. However, the precise delivery of CRISPR-based therapies to DCs has posed a significant challenge. While lipid nanoparticles (LNPs) have been extensively studied for gene editing in tumor cells, their potential application in DCs has remained relatively unexplored. This study investigates the important role of cholesterol in regulating the efficiency of BAMEA-O16B lipid-assisted nanoparticles (BLANs) as carriers of CRISPR/Cas9 for gene editing in DCs. Remarkably, BLANs with low cholesterol density exhibit exceptional mRNA uptake, improved endosomal escape, and efficient single-guide RNA release capabilities. Administration of BLAN_mCas9/gPD-L1 results in substantial PD-L1 gene knockout in conventional dendritic cells (cDCs), accompanied by heightened cDC1 activation, T cell stimulation, and significant suppression of tumor growth. The study underscores the pivotal role of cholesterol density within LNPs, revealing potent influence on gene editing efficacy within DCs. This strategy holds immense promise for the field of cancer immunotherapy, offering a novel avenue for treating immune-related diseases.

Objective To search,evaluate and summarise the best evidence on pre-rehabilitation before surgery for elderly patients with hip fracture so as to provide an evidence-based basis for further standardisation in clinical practice.Methods Based on the"6S Pyramid"evidence model,desktop searches were conducted across databases and websites including Medlive,BMJ Best Practice,UpToDate,the World Health Organization,the Guidelines International Network(GIN),British National Institute for Health and Care Excellence(NICE),American National Guideline Clearinghouse(NGC),Canadian Medical Association:Clinical Practice Guidelines Infobase(CMA),Scottish Intercollegiate Guidelines Network(SIGN),the Registered Nurses'Association of Ontario(RNAO),American College of Sports Medicine(ACSM),American Academy of Orthopaedic Surgeons(AAOS),European Society for Clinical Nutrition and Metabolism(ESPEN),American Society for Parenteral and Enteral Nutrition(ASPEN),the Enhanced Recovery After Surgery Society(ERASS),Cochrane Library,Joanna Briggs Institute,PubMed,CINAHL,Web of Science,Embase,Scopus,Science Direct,CNKI,Wanfang Data,Vip,and SinoMed.The research targeted evidence on pre-rehabilitation in elderly patients with hip-fracture before surgery,encompassing clinical decisions,guidelines,evidence summaries,best practices,expert consensus,systematic reviews and randomized controlled trials(RCTs).The search items spanned from the inception of database to 31st December,2023.Two researchers trained with evidence-based knowledge evaluated the quality of the included literature and extracted and summarised the evidence.Results A total of 14 documents were included,consisting of 2 clinical decisions,4 guidelines,1 best practice,4 expert consensus and 3 systematic reviews.Evidence was summarised into 7 domains from 25 pieces of evidence:multidisciplinary management,preoperative assessment,preoperative health education,multi-modal analgesia,nutritional management,exercise intervention and assessment and prevention of complication risks.Conclusion The best evidence summarised in this study provides a basis for pre-rehabilitation in elderly patients with hip fracture before surgery.Nurses should adapt the findings to specific contexts,standardise the preoperative rehabilitation procedures,and improve clinical outcomes for the patients.

Fragility fractures represent the final clinical manifestation of osteoporosis and are influenced by a range of risk factors. This article discusses the findings from two large-scale studies utilizing biobank and genomic data to investigate the relationships among obesity, type 2 diabetes and fracture risk, and to explore their implications for health management. The studies suggest that general obesity(defined by body mass index, BMI), central obesity(defined by waist circumference), and peripheral obesity exert different effects on fracture risk in older adults. Specifically, individuals who are overweight based on BMI have a lower fracture risk compared to those with normal weight, whereas an increased waist circumference is associated with a higher fracture risk. Moreover, genetic research indicates that genetically determined type 2 diabetes is not causally associated with increased fracture risk. In contrast, observational studies have reported a higher fracture risk in patients with type 2 diabetes; however, this association becomes non-significant after adjusting for diabetes-related risk factors. These findings highlight that maintaining a moderately elevated BMI may benefit bone formation in the elderly by providing mechanical loading to stimulate bone formation. Additionally, effective management of type 2 diabetes complications is essential for reducing fracture risk, and a certain degree of obesity may confer a protective effect against fractures in individuals with type 2 diabetes.

The purpose of this study was to prepare high affinity monoclonal antibodies(mAbs)a-gainst bovine viral diarrhea virus(BVDV)and establish a double antibody sandwich ELISA detec-tion method.BVDV was purified by differential ultracentrifugation and used to immunize BALB/c mice.Hybridoma cells were prepared by fusing spleen cells from the immunized mice with SP2/0 cells.Positive cells were screened by indirect ELISA.A double-antibody sandwich ELISA method for detecting BVDV was developed using monoclonal antibody 4D11 as the capture antibody and HRP-labeled monoclonal antibody 3F3 as the detection antibody.The results of the ELISA and the determination of the variable region gene sequence of monoclonal antibodies indicated that the two monoclonal antibodies recognize different antigenic epitopes.Specificity tests showed that two monoclonal antibodies specifically recognize BVDV and did not cross-react with other bovine viru-ses associated with diarrhea.Indirect immunofluorescence assay and Western blot assay demonstra-ted that both mAbs exhibited strong reactivity with BVDV.The double antibody sandwich ELISA detection method established in this study had good specificity.The sensitivity test revealed that the method could detect a minimum virus amount of 3.1 × 104 TCID50.The reproducibility test showed that the inter-batch coefficient of variation(Cv)was between 2.47％and 7.44％,and the intra-batch Cv was between 1.71％and 9.89％,indicating good reproducibility.The establishment of this method provides an effective technical tool for the rapid diagnosis and prevention and con-trol of BVDV.

Objective To investigate the effects of transcutaneous electrical nerve stimulation(TENS)combined with mobilization with movement(MWM)on clinical efficacy,musculoskeletal ultrasound findings and bone metabolism indicators in patients with knee osteoarthritis(KOA).Methods Ninety KOA patients were collected and randomly divided into the control group(conventional rehabilitation+TENS,n=45)and the observation group(conventional rehabilitation+TENS+MWM,n=45)using a random number table.Clinical efficacy,knee joint function,musculoskeletal ultrasound indices and bone metabolism indices were compared between the two groups.Results The clinical efficacy of the observation group was superior to that of the control group(P＜0.05).At 2 and 6 months post-treatment,Western Ontario and McMaster Universities Osteoarthritis Index(WOMAC)were significantly lower in the observation group than those of the control group,and Lysholm Knee Scoring Scale scores were higher in the observation group than those of the control group(P＜0.05).Additionally,articular cartilage thickness,bone gla protein(BGP)and bone-specific alkaline phosphatase(BALP)were higher in the observation group than those of the control group,and synovial thickness,suprapatellar effusion and tartrate-resistant acid phosphatase 5b(TRACP-5b)were lower in the observation group than those of the control group(P＜0.05).Conclusion TENS combined with MWM in the treatment of KOA patients can not only improve knee joint function but also optimize musculoskeletal ultrasound indices and regulate bone metabolism indices.

The purpose of this study was to prepare high affinity monoclonal antibodies(mAbs)a-gainst bovine viral diarrhea virus(BVDV)and establish a double antibody sandwich ELISA detec-tion method.BVDV was purified by differential ultracentrifugation and used to immunize BALB/c mice.Hybridoma cells were prepared by fusing spleen cells from the immunized mice with SP2/0 cells.Positive cells were screened by indirect ELISA.A double-antibody sandwich ELISA method for detecting BVDV was developed using monoclonal antibody 4D11 as the capture antibody and HRP-labeled monoclonal antibody 3F3 as the detection antibody.The results of the ELISA and the determination of the variable region gene sequence of monoclonal antibodies indicated that the two monoclonal antibodies recognize different antigenic epitopes.Specificity tests showed that two monoclonal antibodies specifically recognize BVDV and did not cross-react with other bovine viru-ses associated with diarrhea.Indirect immunofluorescence assay and Western blot assay demonstra-ted that both mAbs exhibited strong reactivity with BVDV.The double antibody sandwich ELISA detection method established in this study had good specificity.The sensitivity test revealed that the method could detect a minimum virus amount of 3.1 × 104 TCID50.The reproducibility test showed that the inter-batch coefficient of variation(Cv)was between 2.47％and 7.44％,and the intra-batch Cv was between 1.71％and 9.89％,indicating good reproducibility.The establishment of this method provides an effective technical tool for the rapid diagnosis and prevention and con-trol of BVDV.

Objective To investigate the effects of transcutaneous electrical nerve stimulation(TENS)combined with mobilization with movement(MWM)on clinical efficacy,musculoskeletal ultrasound findings and bone metabolism indicators in patients with knee osteoarthritis(KOA).Methods Ninety KOA patients were collected and randomly divided into the control group(conventional rehabilitation+TENS,n=45)and the observation group(conventional rehabilitation+TENS+MWM,n=45)using a random number table.Clinical efficacy,knee joint function,musculoskeletal ultrasound indices and bone metabolism indices were compared between the two groups.Results The clinical efficacy of the observation group was superior to that of the control group(P＜0.05).At 2 and 6 months post-treatment,Western Ontario and McMaster Universities Osteoarthritis Index(WOMAC)were significantly lower in the observation group than those of the control group,and Lysholm Knee Scoring Scale scores were higher in the observation group than those of the control group(P＜0.05).Additionally,articular cartilage thickness,bone gla protein(BGP)and bone-specific alkaline phosphatase(BALP)were higher in the observation group than those of the control group,and synovial thickness,suprapatellar effusion and tartrate-resistant acid phosphatase 5b(TRACP-5b)were lower in the observation group than those of the control group(P＜0.05).Conclusion TENS combined with MWM in the treatment of KOA patients can not only improve knee joint function but also optimize musculoskeletal ultrasound indices and regulate bone metabolism indices.

Objective To search,evaluate and summarise the best evidence on pre-rehabilitation before surgery for elderly patients with hip fracture so as to provide an evidence-based basis for further standardisation in clinical practice.Methods Based on the"6S Pyramid"evidence model,desktop searches were conducted across databases and websites including Medlive,BMJ Best Practice,UpToDate,the World Health Organization,the Guidelines International Network(GIN),British National Institute for Health and Care Excellence(NICE),American National Guideline Clearinghouse(NGC),Canadian Medical Association:Clinical Practice Guidelines Infobase(CMA),Scottish Intercollegiate Guidelines Network(SIGN),the Registered Nurses'Association of Ontario(RNAO),American College of Sports Medicine(ACSM),American Academy of Orthopaedic Surgeons(AAOS),European Society for Clinical Nutrition and Metabolism(ESPEN),American Society for Parenteral and Enteral Nutrition(ASPEN),the Enhanced Recovery After Surgery Society(ERASS),Cochrane Library,Joanna Briggs Institute,PubMed,CINAHL,Web of Science,Embase,Scopus,Science Direct,CNKI,Wanfang Data,Vip,and SinoMed.The research targeted evidence on pre-rehabilitation in elderly patients with hip-fracture before surgery,encompassing clinical decisions,guidelines,evidence summaries,best practices,expert consensus,systematic reviews and randomized controlled trials(RCTs).The search items spanned from the inception of database to 31st December,2023.Two researchers trained with evidence-based knowledge evaluated the quality of the included literature and extracted and summarised the evidence.Results A total of 14 documents were included,consisting of 2 clinical decisions,4 guidelines,1 best practice,4 expert consensus and 3 systematic reviews.Evidence was summarised into 7 domains from 25 pieces of evidence:multidisciplinary management,preoperative assessment,preoperative health education,multi-modal analgesia,nutritional management,exercise intervention and assessment and prevention of complication risks.Conclusion The best evidence summarised in this study provides a basis for pre-rehabilitation in elderly patients with hip fracture before surgery.Nurses should adapt the findings to specific contexts,standardise the preoperative rehabilitation procedures,and improve clinical outcomes for the patients.

Fragility fractures represent the final clinical manifestation of osteoporosis and are influenced by a range of risk factors. This article discusses the findings from two large-scale studies utilizing biobank and genomic data to investigate the relationships among obesity, type 2 diabetes and fracture risk, and to explore their implications for health management. The studies suggest that general obesity(defined by body mass index, BMI), central obesity(defined by waist circumference), and peripheral obesity exert different effects on fracture risk in older adults. Specifically, individuals who are overweight based on BMI have a lower fracture risk compared to those with normal weight, whereas an increased waist circumference is associated with a higher fracture risk. Moreover, genetic research indicates that genetically determined type 2 diabetes is not causally associated with increased fracture risk. In contrast, observational studies have reported a higher fracture risk in patients with type 2 diabetes; however, this association becomes non-significant after adjusting for diabetes-related risk factors. These findings highlight that maintaining a moderately elevated BMI may benefit bone formation in the elderly by providing mechanical loading to stimulate bone formation. Additionally, effective management of type 2 diabetes complications is essential for reducing fracture risk, and a certain degree of obesity may confer a protective effect against fractures in individuals with type 2 diabetes.

Background and purpose:Chimeric antigen receptor T(CAR-T)cell therapy has shown remarkable efficacy in treating hematological and lymphatic system tumors,but its effectiveness in solid tumors is relatively poor,which is partly attributed to target selection.For Ewing sarcoma(ES),CD99 can be a potential target for CAR-T cells.However,due to T cells'endogenous expression of CD99 protein,CAR-T cells targeting CD99 face limitations in their expansion capacity in vitro.This study aimed to identify the optimal conditions for preparing CD99 CAR-T cells by incorporating CD99 knockdown short hairpin RNA(shRNA),optimizing the multiplicity of infection(MOI)for lentiviral transduction,and screening for the best culture medium and container for CAR-T cell expansion.Methods:shRNA sequences were screened to enhance the expansion capacity of CD99 CAR-T cells.Different MOI,culture media,and containers were used to assess CAR-T cell transduction efficiency,cell viability,proliferation capacity,specific killing ability,and interferon-γ(IFN-γ)release levels under various conditions,in order to identify the optimal cell preparation conditions.Results:The expansion level of KO-CD99 CAR-T cells obtained through shRNA knockdown was significantly higher than that of CD99 CAR-T cells[(16.40±0.40)vs(6.33±1.53),P<0.01].The optimal expansion effect was observed when the transduction MOI was between 0.25 and 1.0,and OptiVitro was used as the culture medium.CAR-T cells cultured in ventilated flasks exhibited significantly higher expansion rates compared to cells cultured in bags[MOI=0.25:(50.23±3.32)vs(13.02±4.82);MOI=0.50:(49.96±0.83)vs(18.25±2.88);MOI=1.00:(48.27±5.08)vs(13.16±6.26);P<0.01],with better cell phenotype and higher specific killing ability.Conclusion:KO-CD99 CAR-T cells obtained through shRNA technology can achieve stable expansion.Based on the optimization of expansion conditions,KO-CD99 CAR-T cells exhibit superior expansion capacity and a higher proportion of memory T cells when the MOI is between 0.25 and 1.00,OptiVitro is used as the culture medium,and ventilated flasks are used as the culture container.These findings lay a solid foundation for further clinical trials of CD99 CAR-T cell therapy for ES.