BACKGROUND:Sarcopenia is an age-related systemic skeletal muscle disease,which is associated with a variety of adverse outcomes such as falls,functional decline,frailty,and death.Postmenopausal women are one of the high-risk groups for sarcopenia.OBJECTIVE:To develop a predictive model for assessing the risk of sarcopenia in Chinese postmenopausal women based on high-quality database.METHODS:Data for this study were derived from 2 370 postmenopausal women from the China Health and Retirement Longitudinal Study(CHARLS),and sarcopenia was assessed using the Asian Working Group on Sarcopenia 2019(AWGS2019)recommended metrics.The study cohort was randomized into a training set(70％)and a validation set(30％).Risk factors for sarcopenia in postmenopausal women were screened using the least absolute shrinkage and selection operator,ten-fold cross-validation,and logistic regression.Nomogram predicting the risk of sarcopenia in postmenopausal women was constructed based on the risk factors,and the model efficacy was evaluated by the receiver operating characteristic curve and area under the curve(AUC),calibration curve,and decision curve analysis.RESULTS AND CONCLUSION:The prevalence of sarcopenia in this study was 23.50％and age,place of residence,sleep quality,cognitive function,depression,and the number of chronic diseases were selected as predictors of sarcopenia in postmenopausal women.The nomogram model showed good discrimination between the training and validation sets,with an AUC value of 0.751(95％confidence interval=0.724-0.778,P＜0.001),a specificity of 72.2％,and a sensitivity of 63.2％in the training set,and an AUC value of 0.763(95％confidence interval=0.721-0.805,P＜0.001),with a specificity of 69.6％and a sensitivity of 70.8％.The calibration curve showed a relatively significant agreement between the nomogram model and the actual observations,and the decision curve analysis demonstrated broad and good clinical utility.To conclude,the nomogram to assess the risk of sarcopenia constructed based on age,place of residence,sleep quality,cognitive function,depression,and number of chronic diseases,provides an effective tool for identifying and eliminating risk factors for sarcopenia in Chinese postmenopausal women,and helps to reduce the incidence of sarcopenia.

BACKGROUND:Sarcopenia is an age-related systemic skeletal muscle disease,which is associated with a variety of adverse outcomes such as falls,functional decline,frailty,and death.Postmenopausal women are one of the high-risk groups for sarcopenia.OBJECTIVE:To develop a predictive model for assessing the risk of sarcopenia in Chinese postmenopausal women based on high-quality database.METHODS:Data for this study were derived from 2 370 postmenopausal women from the China Health and Retirement Longitudinal Study(CHARLS),and sarcopenia was assessed using the Asian Working Group on Sarcopenia 2019(AWGS2019)recommended metrics.The study cohort was randomized into a training set(70％)and a validation set(30％).Risk factors for sarcopenia in postmenopausal women were screened using the least absolute shrinkage and selection operator,ten-fold cross-validation,and logistic regression.Nomogram predicting the risk of sarcopenia in postmenopausal women was constructed based on the risk factors,and the model efficacy was evaluated by the receiver operating characteristic curve and area under the curve(AUC),calibration curve,and decision curve analysis.RESULTS AND CONCLUSION:The prevalence of sarcopenia in this study was 23.50％and age,place of residence,sleep quality,cognitive function,depression,and the number of chronic diseases were selected as predictors of sarcopenia in postmenopausal women.The nomogram model showed good discrimination between the training and validation sets,with an AUC value of 0.751(95％confidence interval=0.724-0.778,P＜0.001),a specificity of 72.2％,and a sensitivity of 63.2％in the training set,and an AUC value of 0.763(95％confidence interval=0.721-0.805,P＜0.001),with a specificity of 69.6％and a sensitivity of 70.8％.The calibration curve showed a relatively significant agreement between the nomogram model and the actual observations,and the decision curve analysis demonstrated broad and good clinical utility.To conclude,the nomogram to assess the risk of sarcopenia constructed based on age,place of residence,sleep quality,cognitive function,depression,and number of chronic diseases,provides an effective tool for identifying and eliminating risk factors for sarcopenia in Chinese postmenopausal women,and helps to reduce the incidence of sarcopenia.

The nursing experience was summarised over a patient who developed an impending myasthenic crisis following immune checkpoint inhibitors(ICIs)therapy after the surgery for stage-Ⅳ liver cancer.Key nursing points included:close monitoring for early signs of impending myasthenic crisis;set up a multi-disciplinary team to improve respiratory,oropharyngeal and ocular function,control respiratory infection,provide psychological support in order to prevent the patient from developing a manifest myasthenia-gravis crisis(MC);combine with medications to relieve myasthenic symptoms and prevent adverse drug reactions;individualised rehabilitation training to promote recovery of muscular strength.Eventually,the patient did not develop into a MC during hospitalisation(20d)and was well at 3-month after discharge.

BACKGROUND:Osteonecrosis is an orthopedic disease that severely limits joint function,with complex pathogenesis involving multiple risk factors.Cathepsins,as a class of enzymes that play a key role in bone metabolism,are closely related to the proliferation,differentiation of bone cells,and remodeling of the bone matrix.However,previous studies have mostly focused on descriptive analyses,lacking direct evidence of causal relationships.OBJECTIVE:To clarify the potential causal relationship between cathepsins and osteonecrosis and to explore their possible mechanisms by analyzing large-scale sample data from the FinnGen database.METHODS:We obtained osteonecrosis-related data from the FinnGen database,including R9(a total of 359 399 samples:1 385 cases and 358 014 controls)and R10 versions(a total of 392 580 samples:1 543 cases and 391 037 controls).Single nucleotide polymorphisms associated with nine cathepsins(cathepsin B,E,F,G,H,O,S,L2,and Z)were acquired from a previous study(3 301 individuals).Univariate Mendelian randomization,reverse univariate Mendelian randomization,and multivariate Mendelian randomization analyses were conducted using the inverse variance weighted method,MR-Egger method,weighted median method,simple mode method,and weighted mode method.Initially,Mendelian randomization analysis was performed using osteonecrosis data from R9.Additionally,sensitivity analyses were conducted using Cochran's Q test,MR-Egger intercept,MR-PRESSO global test,and leave-one-out analysis to check for horizontal pleiotropy and heterogeneity.Subsequently,a validation analysis study was carried out on the R10 dataset,and a meta-analysis was conducted to combine the two datasets to explore the joint effect.RESULTS AND CONCLUSION:Univariate Mendelian randomization analysis results showed that higher levels of cathepsin B were significantly associated with a reduced risk of osteonecrosis(inverse variance weighted:odds ratio(OR)=0.865,95％confidence interval(CI):0.762-0.982,P=0.025),and no reverse causal relationship was found between the nine cathepsins and osteonecrosis(P＞0.05).These associations were validated by meta-analysis.Multivariate analysis,using the nine cathepsins as covariates,revealed a reverse causal relationship between the levels of cathepsin Band the risk of osteonecrosis(inverse variance weighted:OR=0.8710,95％CI:0.761-0.997,P=0.045),consistent with the results before adjustment.Sensitivity analyses based on heterogeneity and horizontal pleiotropy suggested that the results were relatively robust.This study suggests that there is a causal relationship between high levels of cathepsin B and the reduced risk of osteonecrosis,and it may serve as a biomarker for osteonecrosis,providing new directions and insights for the diagnosis and treatment of osteonecrosis.Although this study is based on data analysis of European populations,these findings have important implications for Chinese biomedical research,especially in understanding disease mechanisms,developing biomarkers,and formulating treatment strategies.They also encourage similar studies conducted on Chinese populations to explore the impact of racial and genetic background differences on the occurrence of osteonecrosis.

Objective:To construct a sizeable naive human Fab phage display antibody library to screen high-affinity specific antibodies in vitro. Methods:Total RNA was extracted from peripheral blood mononuclear cells (PBMCs) of 126 healthy individuals, subsequently reverse-transcribed into cDNA, and used as a template. PCR amplification was performed to obtain the V H from IgG, IgM and light chain κ, λ, separately, with the initial PCR products serving as templates for a second round of PCR. Overlap extension PCR was employed to generate fragments of the κ and λ light chains. These fragments were ligated with the phage vector pNC3, which harbors the variable region 1 of the heavy chain, to construct a recombinant phage plasmid. This plasmid was then electroporated into competent Escherichia Coli TG1 cells to establish a naive human Fab phage display antibody library. One hundred clones were randomly selected for identification and sequencing, and antibody gene polymorphisms were analyzed using the IMGT database and MAFFT software. Recombinant α-hemolysin from Staphylococcus aureus was utilized to screen Fab antibody fragments through biopanning of the antibody library, followed by random selection of phage ELISA-identified clones. The positive clones (antigen A450∶blank control A450≥2.1) were sequenced. Results:Two large naive Fab phage display antibody libraries were successfully constructed, in which the capacity of κ and λ chain antibody libraries were 1.25×10 11 and 1.54×10 11, respectively. The titers for two antibody libraries were 6.04×10 13 CFU/ml and 3.50×10 13 CFU/ml. The positive transformation insertion rates for κ and λ chain antibody libraries were 96% (96/100) and 100% (100/100), respectively. Sequence analysis revealed that all antibody sequences were unique. The amino acid sequences in the skeletal region were relatively conserved. In contrast, significant variations in the length of the complementarity determining region (CDR) were found, and the diversity of amino acid sequence of the complementary determining region was high, especially the CDR3. Analysis using the IMGT database indicated that the sequences exhibited a broad distribution across variable-diversity-joining gene families. After six rounds of panning, specific phage antibodies enrichment targeting α-hemolysin were achieved. A total of 142 monoclonal antibodies were sequenced, yielding 8 distinct Fab antibody sequences. Conclusion:This study successfully constructed two naive human Fab phage display antibody libraries with large capacity and good diversity, which can be used for screening human antibodies for serum epidemiology.

Objective:To construct a sizeable naive human Fab phage display antibody library to screen high-affinity specific antibodies in vitro. Methods:Total RNA was extracted from peripheral blood mononuclear cells (PBMCs) of 126 healthy individuals, subsequently reverse-transcribed into cDNA, and used as a template. PCR amplification was performed to obtain the V H from IgG, IgM and light chain κ, λ, separately, with the initial PCR products serving as templates for a second round of PCR. Overlap extension PCR was employed to generate fragments of the κ and λ light chains. These fragments were ligated with the phage vector pNC3, which harbors the variable region 1 of the heavy chain, to construct a recombinant phage plasmid. This plasmid was then electroporated into competent Escherichia Coli TG1 cells to establish a naive human Fab phage display antibody library. One hundred clones were randomly selected for identification and sequencing, and antibody gene polymorphisms were analyzed using the IMGT database and MAFFT software. Recombinant α-hemolysin from Staphylococcus aureus was utilized to screen Fab antibody fragments through biopanning of the antibody library, followed by random selection of phage ELISA-identified clones. The positive clones (antigen A450∶blank control A450≥2.1) were sequenced. Results:Two large naive Fab phage display antibody libraries were successfully constructed, in which the capacity of κ and λ chain antibody libraries were 1.25×10 11 and 1.54×10 11, respectively. The titers for two antibody libraries were 6.04×10 13 CFU/ml and 3.50×10 13 CFU/ml. The positive transformation insertion rates for κ and λ chain antibody libraries were 96% (96/100) and 100% (100/100), respectively. Sequence analysis revealed that all antibody sequences were unique. The amino acid sequences in the skeletal region were relatively conserved. In contrast, significant variations in the length of the complementarity determining region (CDR) were found, and the diversity of amino acid sequence of the complementary determining region was high, especially the CDR3. Analysis using the IMGT database indicated that the sequences exhibited a broad distribution across variable-diversity-joining gene families. After six rounds of panning, specific phage antibodies enrichment targeting α-hemolysin were achieved. A total of 142 monoclonal antibodies were sequenced, yielding 8 distinct Fab antibody sequences. Conclusion:This study successfully constructed two naive human Fab phage display antibody libraries with large capacity and good diversity, which can be used for screening human antibodies for serum epidemiology.

BACKGROUND:Previous studies by the research group have shown that core proteoglycan in Cornus Cervi Col la can enter the bone,promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells,and has a good repair effect on steroid-induced osteonecrosis of the femoral head.OBJECTIVE:To investigate the therapeutic effect and potential mechanism of Cornus Cervi Colla on steroid-induced osteonecrosis of the femoral head in rats by fecal non-targeted fecal metabolomics.METHODS:Thirty SD rats were randomly divided into three groups using a random number table method:the control group(n=10)was injected with normal saline into the right gluteal muscle(injected on the first 3 days of each week for 3 consecutive weeks),and was given pure water gavage(once a day for 6 consecutive weeks).The model group(n=10)was injected with methylprednisolone sodium succinate into the right gluteal muscle(injected on the first 3 days of each week for 3 consecutive weeks)to establish a steroid-induced osteonecrosis of the femoral head model,and was given pure water gavage(once a day for 6 consecutive weeks).The Cornus Cervi Co I la group(n=10)was also established with a steroid-induced osteonecrosis of the femoral head model,and was given Cornus Cervi Col la gavage(once a day for 6 consecutive weeks).After gavage,cecal feces and femoral heads were collected for fecal metabolomics analysis and bone tissue Micro-CT and hematoxylin-eosin staining,respectively.RESULTS AND CONCLUSION:(1)Metabolomics analysis results showed that there were 233 differential metabolites between the Cornus Cervi Col la and model groups,with 65 significantly differing and clearly annotated metabolites.Lipid and amino acid metabolites were significantly increased,with bile acids,sulfated steroids,ephedrine,hypoxanthine,betaine,L-carnitine,B-mouse bile acid,cytidine,4-pyridoxic acid,taurine,N-acetyl-d-glucosamine,and butyric acid being the most impacted(variable weight value VIP＞5).The metabolic pathways of taurine and hypotaurine were crucial in the metabolic regulatory network(pathway impact=0.428 57).(2)Micro-CT scanning results of bone tissue showed that the femoral heads of rats in the model group and the Cornus Cervi Col la group had different degrees of damage;the femoral head contour was irregular;the trabeculae in the subchondral bone of the femoral head were missing and disordered,and some cystic structures were visible.Compared with the model group,the degree of trabecular damage in the rats of the Cornus Cervi Colla group was milder.Hematoxylin-eosin staining results showed that the trabeculae in the subchondral bone of the femoral heads of rats in the model group and the Cornus Cervi Colla group were sparse or interrupted,and the empty bone lacuna rate and adipocyte invasion were increased.Compared with the model group,the empty bone lacuna rate and adipocyte invasion in the subchondral bone of the femoral heads of rats in the Cornus Cervi Colla group were reduced.(3)These results conclude that Cornus Cervi Colla potentially mitigates steroid-induced osteonecrosis of the femoral head through the metabolic processes involving taurine and associated pathways.

ObjectiveTo investigate the application of the novel inflammatory factor adsorption column CA280 combined with low-dose plasma exchange （LPE） in patients with acute-on-chronic liver failure （ACLF）. MethodsA prospective cohort study was designed， and a total of 93 ACLF patients who were admitted to The Ninth Hospital of Nanchang from June 2023 to January 2025 were enrolled and randomly divided into DPMAS+LPE group with 50 patients and CA280+LPE group with 43 patients. In addition to comprehensive medical treatment， the patients in the DPMAS+LPE group received DPMAS and LPE treatment， and those in the CA280+LPE group received CA280 and LPE treatment. The two groups were observed in terms of routine blood test results， liver function parameters， renal function markers， electrolytes， coagulation function parameters， cytokines， adverse events， and 28-day prognosis before surgery （baseline）， during surgery （DPMAS or CA280）， and after surgery （after sequential LPE treatment）. The paired t-test was used for comparison of normally distributed continuous data before and after treatment within each group， and the independent-samples t test was used for comparison between groups； the Wilcoxon signed-rank test was used for comparison of non-normally distributed continuous data before and after treatment within each group， and the Mann-Whitney U test was used for comparison between groups. The chi-square test or the Fisher’s exact test was used for comparison of categorical data between groups， and the Spearman test was used for correlation analysis. ResultsAfter CA280 treatment， the ACLF patients had significant reductions in the levels of cytokines （IL-6， IL-8， IL-10， TNF-α， and IFN-γ）， liver function parameters （ALT， AST， ALP， TBil， DBil， Alb， and glutathione reductase）， and the renal function marker urea nitrogen （all P<0.05）， and in terms of coagulation function parameters， there were significant increases in prothrombin time， activated partial thromboplastin time （APTT）， thrombin time， and international normalized ratio （INR） and significant reductions in prothrombin activity （PTA） and fibrinogen （FIB） （all P<0.05）. Compared with the DPMAS+LPE group， the CA280+LPE group showed better improvements in the serum cytokines IL-8 （Z=-2.63， P=0.009）， IL-10 （Z=-3.94， P<0.001）， and TNF-α （Z=-1.53， P=0.023）， and the two artificial liver support systems had a similar effect in improving liver function （ALT， AST， GGT， GR， TBil， and DBil） （all P >0.05）， but the CA280+LPE group showed a significantly greater reduction in Alb （Z=-2.08， P=0.037）. CA280+LPE was more effective in reducing uric acid （Z=-2.97， P=0.003）. Compared with DPMAS+LPE， CA280+LPE treatment resulted in a significant reduction in INR （Z=-4.01， P<0.001）， a significant increase in APTT （Z=-2.53， P=0.011）， and significant greater increases in PTA （Z=-6.28， P<0.001） and FIB （Z=-3.93， P<0.001）. There were no significant differences in the incidence rates of adverse reactions and the rate of improvement at discharge between the two groups （all P>0.05）. The Spearman correlation analysis showed that IL-6 was significantly correlated with WBC （r=0.22， P=0.042）， TBil （r=0.29， P=0.005）， and FIB （r=-0.33， P=0.003）； IL-8 was positively correlated with APTT （r=0.37， P<0.001） and INR （r=0.25， P=0.013）； TNF-α was significantly correlated with WBC （r=0.40， P<0.001） and TBil （r=0.34， P<0.001）. ConclusionCompared with DPMAS， CA280 combined with LPE can effectively clear proinflammatory cytokines and improve liver function in ACLF patients， but it has a certain impact on Alb and coagulation function. This regimen provides a new option for the individualized treatment of ACLF and can improve the short-term prognosis of patients， but further studies are needed to verify its long-term efficacy.

BACKGROUND:With the continuous exploration of the pathogenesis of osteonecrosis,more and more research evidence shows that neuroimaging change is closely related to the onset of osteonecrosis.However,the specific causal relationship between neuroimaging change and osteonecrosis is still unclear.OBJECTIVE:To evaluate the causal relationship between neuroimaging indices and osteonecrosis using Mendelian Randomization analysis.METHODS:Neuroimaging data were obtained from the UK Biobank database in the UK,which included a total of 36 778 individuals.Osteonecrosis data were obtained from the FinnGen database in Finland,including 1 543 cases and 391 037 controls.Instrumental variables were extracted and screened from outcome factors,and two-sample Mendelian randomization analysis was performed.The data were analyzed by inverse variance weighted method,MR-Egger,weighted median method,simple model method,and weighted model method.The inverse variance weighted method was used as the main analysis method,and the other four methods were used as supplements.To verify the feasibility and stability of the data,sensitivity analysis of the results was performed.Based on the complexity of causal inference,a reverse Mendelian randomization analysis was further performed to evaluate the potential reverse causal relationship.RESULTS AND CONCLUSION:(1)The results of inverse variance weighted analysis showed that 97 neuroimaging data were positively correlated with osteonecrosis(P＜0.05,OR＞1);2 data were heterogeneous and 6 data had horizontal pleiotropy.95 neuroimaging phenotypes were negatively correlated with osteonecrosis(P＜0.05,OR＜1);5 data were heterogeneous,and 9 data had horizontal pleiotropy;2 groups of data had reverse causal relationships.(2)The two-sample Mendelian randomization analysis established the causal relationship between neuroimaging indicators and osteonecrosis in the academic community.These large sample numbers from the UK and Finland provide a new theoretical basis for the pathophysiology of osteonecrosis,and also provide ideas and methods for the prediction,screening,early diagnosis and prognosis of osteonecrosis in China,which is conducive to improving the accuracy of clinical diagnosis and the effectiveness of treatment of osteonecrosis.

BACKGROUND:With the continuous exploration of the pathogenesis of osteonecrosis,more and more research evidence shows that neuroimaging change is closely related to the onset of osteonecrosis.However,the specific causal relationship between neuroimaging change and osteonecrosis is still unclear.OBJECTIVE:To evaluate the causal relationship between neuroimaging indices and osteonecrosis using Mendelian Randomization analysis.METHODS:Neuroimaging data were obtained from the UK Biobank database in the UK,which included a total of 36 778 individuals.Osteonecrosis data were obtained from the FinnGen database in Finland,including 1 543 cases and 391 037 controls.Instrumental variables were extracted and screened from outcome factors,and two-sample Mendelian randomization analysis was performed.The data were analyzed by inverse variance weighted method,MR-Egger,weighted median method,simple model method,and weighted model method.The inverse variance weighted method was used as the main analysis method,and the other four methods were used as supplements.To verify the feasibility and stability of the data,sensitivity analysis of the results was performed.Based on the complexity of causal inference,a reverse Mendelian randomization analysis was further performed to evaluate the potential reverse causal relationship.RESULTS AND CONCLUSION:(1)The results of inverse variance weighted analysis showed that 97 neuroimaging data were positively correlated with osteonecrosis(P＜0.05,OR＞1);2 data were heterogeneous and 6 data had horizontal pleiotropy.95 neuroimaging phenotypes were negatively correlated with osteonecrosis(P＜0.05,OR＜1);5 data were heterogeneous,and 9 data had horizontal pleiotropy;2 groups of data had reverse causal relationships.(2)The two-sample Mendelian randomization analysis established the causal relationship between neuroimaging indicators and osteonecrosis in the academic community.These large sample numbers from the UK and Finland provide a new theoretical basis for the pathophysiology of osteonecrosis,and also provide ideas and methods for the prediction,screening,early diagnosis and prognosis of osteonecrosis in China,which is conducive to improving the accuracy of clinical diagnosis and the effectiveness of treatment of osteonecrosis.