Objective To investigate prokaryotic expression of the antigen sequence (amino acids 59-145) of mouse leukemia-related protein 16 (LRP16) protein and preparation of rabbit anti-mouse LRP16 polyclonal antibody. Methods The prokaryotic expression plasmid pLS962-LRP16 was constructed by the molecular cloning method and transferred into E.coli Rosetta to express LRP16 protein induced by IPTG. The recombinant protein was purified using Ni-NTA affinity columns followed by gel filtration chromatography. New Zealand white rabbits were immunized with the purified antigen to generate polyclonal antiserum, with antibody titer quantified by ELISA. Antigen-specific IgG was affinity-purified using Sepharose-coupled LRP16 and validated through Western blot and immunofluorescence assays. Results SDS-PAGE analysis confirmed insoluble expression of the LRP16 fusion protein as inclusion bodies. ELISA demonstrated exceptional antiserum titer (1:256 000). Western blot and immunofluorescence verified that the polyclonal antibody could specifically recognize endogenous LRP16 in murine tissues. Conclusion The prokaryotic expression of the LRP16 gene is successfully achieved, and the rabbit anti-mouse LRP16 polyclonal antibody exhibiting high specificity is prepared. This lays the foundation for further studies on the function of the LRP16 gene.
Animals ; Rabbits ; Mice ; Antibodies/immunology* ; Escherichia coli/metabolism* ; Enzyme-Linked Immunosorbent Assay ; Blotting, Western ; Antibody Specificity

To investigate the effect of Xuebijing injection (XBJ) on cGAS/STING pathway in alleviating sepsis-induced acute lung injury (ALI), the mouse sepsis-induced ALI model was established by cecal ligation and puncture (CLP), and the cell inflammation model was constructed by LPS stimulating RAW264.7 cells. The effects of XBJ on lung tissue injury and cGAS/STING pathway-related protein expression in septic mice were investigated by HE staining, ELISA, and Western blot. The results showed that XBJ intervention could alleviate lung tissue injury, reduce serum IL-6, TNF-α, IFN-β, IL-1-β levels, and the expression of cGAS, STING, p-TBK1, and p-IRF3 proteins in lung tissue in vivo, and reduce the mRNA level of related inflammatory factors in RAW264.7 cells and the expression of cGAS/STING pathway proteins in vitro. The results showed that XBJ could play a role in the prevention and treatment of sepsis-induced ALI by inhibiting the inflammatory response via inhibition of the activation of cGAS/STING pathway. This study provides a new molecular mechanism for the clinical prevention and treatment of sepsis-induced acute lung injury with XBJ.

ObjectiveThis study aimed to investigate the intervention effect of Renqing Mangjue on the malignant transformation of gastric mucosal epithelial cells induced by N-methyl-N′-nitro-N-nitrosoguanidine （MNNG） and to explore its molecular mechanism in preventing precancerous lesions of gastric cancer based on the cyclic guanosine monophosphate （cGMP）/protein kinase G （PKG）/mitogen-activated protein kinase （MEK）/extracellular signal-regulated kinase （ERK） signaling pathway. MethodsHuman gastric mucosal epithelial cells （GES-1） were initially induced by MNNG to establish a precancerous cell model （MC cells）. The effective concentration of MNNG for inducing malignant transformation in GES-1 cells was screened using the cell proliferation activity decection （CCK-8） assay， and the effective concentration of Renqing Mangjue for inhibiting the proliferation of transformed GES-1 cells was also determined. GES-1 cells were divided into a blank control group， a model group， and treatment groups with Renqing Mangjue at concentrations of 1， 3， 10， and 30 mg·L^-1. Furthermore， the effects of Renqing Mangjue on the migratory ability and epithelial-mesenchymal transition （EMT） characteristics of GES-1 malignant transformed cells were evaluated using Transwell migration assays， wound healing assays， and real-time quantitative reverse transcription polymerase chain reaction （Real-time PCR）. Additionally， candidate chemical components and target sites of Renqing Mangjue were obtained from the TCMIP v2.0 database， and disease targets at various stages of gastric cancer precursors were sourced from the Gene Expression Omnibus （GEO） database. Pathway enrichment analysis was performed using the Metascape database to predict the potential mechanisms of action of Renqing Mangjue. Finally， the protective mechanism of Renqing Mangjue against gastric cancer precursors was validated through Western blot analysis. ResultsAt a concentration of 20 μmol·L^-1， MNNG exhibited an inhibition rate of approximately 50% on GES-1 cells （P<0.01）， and at this concentration， the GES-1 cells displayed biological characteristics indicative of malignant transformation. In contrast， Renqing Mangjue had no significant effect on the proliferation of normal GES-1 cells， but significantly inhibited the proliferation of MC cells （P<0.01） and markedly reduced their migratory capacity （P<0.01）. Moreover， it also increased the mRNA expression level of E-cadherin during the EMT process （P<0.05）， while inhibiting the expression of both N-cadherin and the transcription factor Snail mRNA （P<0.05， P<0.01）. Network predictions suggested that Renqing Mangjue may prevent gastric cancer precursors through modulating the cGMP/PKG and MAPK/ERK signaling pathways. Furthermore， Western blot results indicated that Renqing Mangjue upregulated the expression of PKG and NPRB （B-type natriuretic peptide receptor） proteins in the cGMP/PKG pathway （P<0.01）， while downregulating the expression of the downstream proteins MEK and ERK （P<0.05， P<0.01）. ConclusionIn summary， Renqing Mangjue can prevent gastric cancer precursors by inhibiting the proliferation and migration of malignant transformed GES-1 cells， thereby delaying the EMT process. The underlying mechanisms may be related to the activation of the cGMP/PKG pathway and the inhibition of the MEK/ERK signaling pathway.

OBJECTIVE:This paper collects relevant literature on blood flow restriction training combined with resistance training,and analyzes the different effects of blood flow restriction training combined with resistance training on athletes'muscle-related indexes and specialized abilities in accordance with the paradigm of systematic evaluation and Meta-analysis,aiming to provide data support for athletes to utilize blood flow restriction training in their training practices. METHODS:Chinese and foreign databases(CNKI,WanFang,PubMed,Web of Science,SPORTDiscus)were searched to collect randomized controlled trials on the effects of blood flow restriction training combined with resistance training on limb circumference,muscle mass,muscle strength,and specialized ability of college athletes from January 1st,2000 to October 12th,2023.At least two researchers evaluated the quality of the included literature using the Cochrane Collaboration risk of bias assessment tools and criteria.Heterogeneity tests,data merging,subgroup analyses,forest plotting,and sensitivity analyses were performed using RevMan 5.4,and funnel plots with publication bias evaluation and sensitivity analyses were performed.The evaluation indexes were limb circumference,muscle thickness,muscle strength and specialized ability,and subgroup analyses were performed for different specialized athletic abilities. RESULTS:(1)A total of 18 randomized controlled trials with 403 subjects were included,and according to the Cochrane Collaboration's risk of bias assessment tool,the quality of literature in the included literature was grade A in 16 articles and grade B in 2 articles.(2)Comparing the effects of blood flow restriction training combined with resistance training and resistance training,there was no significant difference between the two groups in terms of limb circumference[standardized mean difference(SMD)=0.03,95%confidence interval:-0.16-0.21,P=0.78],and a significant difference between the two groups in terms of muscle thickness(SMD=0.14,95%CI:0.01-0.27,P=0.03)and muscle strength(SMD=0.37,95%CI:0.14-0.60,P=0.001).(3)Subgroup analyses of the indicators of specialized capacity indicated that there was high heterogeneity in the analyzed results of distance metrics(I2=73%)and time metrics(I2=55%),which was analyzed as a possible reason due to the differences in testing methods and assessment of metrics'significance in the studies;there was no heterogeneity(I2=0%)in the analyzed results of power metrics;blood flow restriction training combined with resistance training had a significant effect on distance metrics(P<0.01).(4)The results of the combined effect showed the effect of blood flow restriction training combined with resistance training vs.resistance training for specialized ability(P=0.41),suggesting that there is no significant effect of different training methods on specialized ability. CONCLUSION:Both blood flow restriction training combined with resistance training and resistance training can promote muscle thickness,muscle strength and specialized ability in college athletes.Meanwhile,blood flow restriction training combined with resistance training has a more significant effect on muscle thickness,muscle strength and some specialized abilities compared with resistance training.Therefore,blood flow restriction training can be scientifically and rationally integrated into specialized training,so as to achieve a better training effect by integrating the differentiated physiological stimuli to the muscles.However,due to the small number of included studies and other possible limitations,more high-quality,multi-sport type and sex randomized trials need to be included in the future to confirm this.

BACKGROUND:Bone marrow mesenchymal stem cells are one of the sources of adipocytes and express all renin-angiotensin system components,but the effect of angiotensin Ⅱ on bone marrow mesenchymal stem cell differentiation into brown adipose tissue is not clear.OBJECTIVE:To observe the effect of angiotensin Ⅱ on bone marrow mesenchymal stem cell differentiation into brown adipose tissue and investigate the role of angiotensin 1a receptor knockout in effect of angiotensin Ⅱ on bone marrow mesenchymal stem cell differentiation into brown adipocytes and its potential mechanisms.METHODS:After isolation and culture of bone marrow mesenchymal stem cells in wild-type and angiotensin 1a receptor knockout SD rats,the cells were cultured to the third generation and randomly divided into four groups:wild type group,knockout group,wild type+angiotensin Ⅱ group,and knockout+angiotensin Ⅱ group.The differentiation was induced in the brown fat induced differentiation medium for 14 days.Angiotensin Ⅱ(100 nmol/L)was added for intervention when the differentiation medium was changed each time in the latter two groups.Western blot assay,qRT-PCR,immunofluorescence,and other methods were used to detect the expression of induced differentiation,lipolysis,β oxidation,and mitochondrial biogenesis in brown fat.RESULTS AND CONCLUSION:Angiotensin Ⅱ could inhibit the browning of rat bone marrow mesenchymal stem cells.Knockout of angiotensin 1a receptor could improve the inhibitory effect of angiotensin Ⅱ on brown lipid formation of rat bone marrow mesenchymal stem cells by promoting lipolysis,enhancing fatty acid β oxidation,promoting mitochondrial biogenesis,and enhancing mitochondrial function.These findings provide new research directions and potential therapeutic targets for obesity treatment,revealing the important role of renin angiotensin systems in fat metabolism and its potential as a therapeutic target.

Ripple structure is widely used in space suits,and the predicted trajectory motion is realized according to its large deformation behavior.Due to the complexity of irregular bellows structure,the existing researches are mostly based on the display dynamic method,and lack the systematic analysis of geometric nonlinearity and the effect of shell section thickness under the framework of statics.This study combines statics and arc length method(Riks),introduces linear buckling mode defects,and analyzes the ultimate bending behavior of irregular bellows with different shell thicknesses.It is concluded that the thick-shell model exhibits higher apparent deformation tolerance due to geometric softening effect,and the contact force distribution between ripples can be accurately extracted by static analysis,which provides a basis for the inversion of external force loads.The deformation mode of bellows after introducing defects based on statics-Riks is proposed,and the control mechanism of shell section thickness on deformation path is revealed.

Objective To investigate the current status and barriers to implementing non-accompanied care services in medical institutions in Zhejiang Province,and to provide a basis for the improvement of standardized management of such services.Methods A self-designed questionnaire was conducted from September to October 2024 among all secondary and tertiary medical institutions in Zhejiang Province to assess the implementation status and barriers to non-accompanied care services.Results A total of 397 questionnaires were distributed,with 389 valid responses,yielding a valid response rate of 97.98％.Non-accompanied care services were implemented in 118 institutions(30.33％).Among these,90 institutions(76.27％)had established management systems for non-accompanied wards;71 institutions(60.17％)had a medical nursing assistant-to-bed ratio lower than 1∶5;41 institu-tions(34.75％)provided tiered training for medical nursing assistants;93 institutions(78.81％)required patients to bear the full cost of the service.Compared with secondary medical institutions,tertiary medical institutions have more complete management system for non-accompanied care services.The main obstacles hindering the development of non-accompanied care services include an imperfect management system for non-accompanied wards,a shortage of medical nursing assistants,a lack of standardized training for such assistants,inconsistent charging standards,and low acceptance among patients and their families.Conclusion The promotion of non-accompanied care services in medical institutions in Zhejiang Province has achieved initial success.However,challenges persist,including incomplete management systems,uneven development across hospital tiers,and imperfect charging mechanisms.It is recommended that relevant authorities strengthen policy support,enhance standardized training for healthcare nursing assistants,refine cost-sharing mechanisms,and improve the quality and sustainability of non-accompanied care services through multi-party collaboration.

Objective:To understand the current situation of the ability to screen health information of the elderly in rural areas and analyze its influencing factors, so as to provide a basis for taking targeted measures.Methods:A cross-sectional survey was adopted, and rural elderly people in Yonghe Town, Dahu Town and Dayao Town of Liuyang City, Hunan Province were selected as the survey subjects by the convenience sampling method. The investigation was conducted using the general information Questionnaire, the Health Information Discrimination Ability Scale, the filial Piety Expectation Scale and the two-way Social Support Scale.Results:A total of 228 questionnaires were distributed, and 215 valid questionnaires were retrieved, with an effective recovery rate of 94.30%. Among the 215 rural elderly people surveyed, 86 were male and 129 were female. The ability to discriminate health information was (69.19 ± 13.41) points, the expectation of filial piety was (37.67 ± 6.12) points, and the two-way social support was (46.82 ± 8.11) points. The ability of rural elderly people to discriminate health information was positively correlated with filial piety expectations and two-way social support ( r = 0.340, 0.456; both P<0.01). The results of hierarchical regression analysis showed that educational level, filial piety expectations and two-way social support were the influencing factors of the current status of health information discrimination ability of rural elderly people ( t values were 2.60 to 5.35, all P<0.05). Conclusions:The ability of discriminating health information of the elderly in rural areas is affected by the educational level, filial piety expectation and two-way social support. The health information of rural elderly people with low education level should be screened and targeted, so as to enrich their cultural life and combine education with fun. Actively build a filial piety culture to promote children′s attention to their parents. In addition to providing social support for the elderly, we should promote their interpersonal and social communication and exert their self-value, so as to improve the health information screening ability of the elderly in rural areas.

Objective:To understand healthcare professionals' perceptions and experiences of early rehabilitation exercise for extracorporeal membrane oxygenation (ECMO) patients, explore existing problems in the implementation of early rehabilitation exercise, and provide reference for constructing safe and effective early rehabilitation exercise programs for ECMO patients.Methods:This study was a qualitative study. Using purposeful sampling method, 13 ICU healthcare professionals (including doctors, rehabilitation therapists, and nurses) from three Class Ⅲ Grade A hospitals in Zhejiang Province that have implemented early rehabilitation exercise for ECMO patients were selected for semi-structured interviews from March to May 2024, and the interview data were collated and analyzed using the Colaizzi 7-step analysis method.Results:The perceptions and experiential perspectives of healthcare professionals on early rehabilitation exercise for ECMO patients can be summarized into three themes and 10 subthemes. The three themes were: healthcare professionals affirm the role of early rehabilitation exercise; lack of unified operational guidelines for early rehabilitation exercise; and patients and their families show high enthusiasm for participating in early rehabilitation exercise.Conclusions:Hospitals should regularly conduct related training on early rehabilitation exercise to enhance the practical skills of healthcare professionals. At the same time, healthcare professionals should focus on utilizing the positive role of patients and their families in the process of early rehabilitation exercise to improve the quality of ECMO patient rehabilitation care.

Objective To investigate the current status and barriers to implementing non-accompanied care services in medical institutions in Zhejiang Province,and to provide a basis for the improvement of standardized management of such services.Methods A self-designed questionnaire was conducted from September to October 2024 among all secondary and tertiary medical institutions in Zhejiang Province to assess the implementation status and barriers to non-accompanied care services.Results A total of 397 questionnaires were distributed,with 389 valid responses,yielding a valid response rate of 97.98％.Non-accompanied care services were implemented in 118 institutions(30.33％).Among these,90 institutions(76.27％)had established management systems for non-accompanied wards;71 institutions(60.17％)had a medical nursing assistant-to-bed ratio lower than 1∶5;41 institu-tions(34.75％)provided tiered training for medical nursing assistants;93 institutions(78.81％)required patients to bear the full cost of the service.Compared with secondary medical institutions,tertiary medical institutions have more complete management system for non-accompanied care services.The main obstacles hindering the development of non-accompanied care services include an imperfect management system for non-accompanied wards,a shortage of medical nursing assistants,a lack of standardized training for such assistants,inconsistent charging standards,and low acceptance among patients and their families.Conclusion The promotion of non-accompanied care services in medical institutions in Zhejiang Province has achieved initial success.However,challenges persist,including incomplete management systems,uneven development across hospital tiers,and imperfect charging mechanisms.It is recommended that relevant authorities strengthen policy support,enhance standardized training for healthcare nursing assistants,refine cost-sharing mechanisms,and improve the quality and sustainability of non-accompanied care services through multi-party collaboration.