Objective:To explore the activation pathway of CD4+CD28-T cells and their specific role in rheumatoid arthritis(RA).Methods:Peripheral blood mononuclear cells(PBMCs)were isolated from RA patients and healthy individuals.Flow cytome-try was used to detect the proportion and activation level of CD4+CD28-T cells,as well as the expressions of perforin and TNF-α,and to analyze the correlation between the proportion of CD4+CD28-T cells and age,C-reactive protein,anti-cyclic citrullinated peptide(CCP)antibodies,rheumatoid factors(RF)and erythrocyte sedimentation rate(ESR).The expression of Toll-like receptor(TLR)2 on CD4+CD28-T cells was detected by flow cytometry.Purified RA CD4+T cells were stimulated with anti-CD3 antibody and TLR2 agonist alone or in combination,and the activation level and cytokine secretion of CD4+CD28-T cells were detected by flow cytometry.Results:The proportion of CD4+CD28-T cells in RA patients was higher than that in healthy individuals(P<0.05),and was positively correlated with rheumatoid factors and erythrocyte sedimentation rate(r=0.554 1,P<0.01;r=0.363 4,P<0.01).Compared with CD4+CD28+T cells,CD25 expression was downregulate(P<0.01),but CD40L and TLR2 expressions were upregulated(P<0.01;P<0.000 1)in CD4+CD28-T cells from RA patients,and they secreted more perforin and TNF-α(P<0.001;P<0.001).After TLR2 ago-nist stimulation,there was no significant change in CD40L and CD25 expression,as well as perforin and TNF-α secretion in CD4+CD28-T cells from RA patients.Conclusion:The proportion of CD4+CD28-T cells is increased in RA patients,and they secrete a large amount of perforin and TNF-α,which may be involved in the inflammatory state and joint damage of RA,but TLR2 cannot re-place CD28 in activating CD4+CD28-T cells.

Objective To develop an enteral nutrition management system for critically ill patients and assess its application outcomes to standardize enteral nutrition management.Methods Based on relevant guidelines and indicator systems,a management system for enteral nutrition in critically ill patients was constructed,consisting of 4 modules:nutritional screening and assessment,nutritional implementation,nutritional monitoring,and statistical analysis.A convenience sampling method was used to select enteral nutrition patients and healthcare staff from the ICU of a tertiary hospital in Ningbo.Data from January to February 2024 served as an experimental group,while data from January to February 2023 constituted a control group.The 2 groups were compared regarding nutritional risk screening rate,feeding interruption rate,completion rate of the enteral nutrition plan,and incidence of complications.At the same time,the system's effectiveness was assessed by healthcare professionals using the clinical nursing information system effectiveness evaluation form.Results The study included 111 patients in the experimental group and 101 patients in the control group.The experimental group exhibited a significantly higher nutritional risk screening rate and enteral nutrition plan completion rate,as well as significantly lower feeding interruption rate and incidence of mechanical complications compared to the control group(P＜0.05).The system received a high effectiveness rating,with an average score of 104.73±9.34.Conclusion The application of the enteral nutrition management system effectively improves the nutritional risk screening rate and completion rate of enteral nutrition plans,while reducing both the feeding interruption rate and the incidence of mechanical complications.Healthcare staff highly rated the system.

Objective To find the relationship between phthalates(PAEs)metabolite concentrations and body mass index(BMI)in infertile patients.Methods From December 2018 to January 2021,120 infertile pa-tients were selected from the Reproductive Medicine Centre of the 988th Hospital of the Joint Logistics Support Force of the Chinese People's Liberation Army and the Reproductive Medicine Centre of the Henan Provincial People's Hospital,while 60 patients with no history of infertility and normal weight were selected from the Medical Check-up Centre as the normal group.According to BMI index,the infertility patients were divided into the control group(BMI＜28 kg/m2),obesity group(obesity,BMI≥28 kg/m2).The obesity group was divided into BHI(BMI≥32 kg/m2)and BH2(BMI≥28 kg/m2,＜32 kg/m2).The basic information of each group was collected.Serum level of PAEs metabolites diethyl phthalate(DEP),dibutyl phthalate(DBP)and di-isooctyl phthalate(DEHP)was detected using liquid chromatography/mass spectrometry(HPLC).The rela-tionship between PAEs metabolite concentration and body mass index(BMI)were analyzed by Spearman's analysis.Results The serum level of DEP,DBP and DEHP was significantly higher in obese group than in the control group(P＜0.05).The serum level of DEP,DBP and DEHP was significantly higher in the control group than in the normal group(P＜0.05);The metabolites level of PAEs was higher in patients with BHI than in the patients with BH2(P＜0.05).Spearman's analysis showed that the BMI of the patients in the obese group of infertility was positively correlated with DEP,DBP and DEHP content(P＜0.05).Conclusions Obesity and PAEs metabolite concentration in infertility patients are related,the higher the degree of obesity,the higher the content of related metabolites.

Objective:To investigate the primary causes of hospitalization, comorbidities, hospitalization costs, and the factors influencing hospitalization expenses in gouty arthritis patients at a tertiary hospital in Haikou, Hainan Province, with the aim of providing a theoretical basis for optimizing medical resource allocation.Methods:We collected hospitalization data from gouty arthritis patients diagnosed between January and December 2021 at a tertiary hospital in Haikou. The main causes of hospitalization, the types and proportions of comorbidities, and their associations with hospitalization costs were analyzed. Patients were categorized into groups based on the primary cause of hospitalization: non-surgical gout, surgery for gout tophiformation, cardiovascular disease, cerebrovascular disease, renal disease, digestive system disease, and type 2 diabetes with complications. Chi-square tests were used for categorical variables, one-way ANOVA or Mann-Whitney U test for continuous variables, and multivariate linear regression for identifying factors influencing hospitalization costs. Spearman′s correlation analysis was conducted to explore associations between variables. Results:A total of 851 patients were included, with 721 males (84.7%) and 130 females (15.3%). The mean age was (65.3±14.1) years, and the average length of stay was (11.5±8.5)days. Among the 833 patients with comorbidities (97.9%), the most prevalent comorbid conditions were hypertension (539 cases, 63.3%), cerebral infarction (378 cases, 44.4%), and type 2 diabetes along with its complications (346 cases, 40.7%). The total cost for the non-surgical gout tophi group was 1.01 (0.72, 1.26) ten thousand yuan, which was significantly lower than that for the gout tophi surgery group [2.04(1.37, 3.42)ten thousand yuan( Z=-4.94, P<0.001)], the cardiovascular disease group [1.30(0.71, 2.29) ten thousand yuan( Z=-4.31, P<0.001)], and the cerebrovascular disease group [1.29(0.98, 2.07) ten thousand yuan, Z=-4.68, P<0.001)]. The total cost for the gout tophi surgery group was 2.04 (1.37, 3.42) ten thousand yuan, which was significantly higher than that for the non-surgical gout tophi group [1.01(0.72, 1.26) ten thousand yuan, Z=-4.94, P<0.001)], the cardiovascular disease group [1.30(0.71, 2.29) ten thousand yuan, Z=-2.40, P=0.020)], the cerebrovascular disease group [1.29(0.98, 2.07) ten thousand yuan, Z=-2.50, P=0.010)], the digestive system disease group [1.04(0.67, 1.97) ten thousand yuan, Z=-2.73, P=0.010)], the renal system disease group [1.01(0.78, 1.53)ten thousand yuan, Z=-3.62, P<0.001)], and the type 2 diabetes and its complications group [(0.88(0.67, 1.25) ten thousand yuan, Z=-4.13, P<0.001)]. Spearman correlation analysis showed a strong positive correlation between length of hospital stay and total hospitalization cost ( r=0.71, P<0.001), as well as a strong positive correlation between total cost and insurance reimbursement ( r=0.87 , P<0.001). The results from multiple linear regression analysis indicated that the length of hospital stay ( B=2 670.36, P<0.001), cardiovascular diseases ( B=12 266.28, P<0.001), digestive system diseases ( B=11 251.76, P=0.001), and cerebrovascular diseases ( B=7 318.79, P=0.020) were significantly positively associated with hospitalization costs. Conclusion:Tophus surgery incurred the highest hospitalization costs, with cardiovascular and cerebrovascular diseases being the primary reasons for gout-related hospitalizations. Standardizing gout treatment may reduce the disease burden by potentially lowering the incidence of comorbidities.

Objectives:To investigate the effect of circular RNA (circRNA) hsa_circ_0019217 on the biological function of chorionic trophoblast cell line (HTR8/SVneo) and its mechanism in the occurrence of pre-eclampsia (PE).Methods:The circRNA expression database was used to analyze the differentially expressed circRNA in placental tissues of PE women. The expression levels of hsa_circ_0019217 and miR-526b-5p were detected by reverse transcription real-time fluorescent quantitative PCR (RT-qPCR), and the subcellular localization of hsa_circ_0019217 was detected by fluorescence in situ hybridization. The proliferation, migration and invasion of trophoblast cells were detected by cell counting kit-8 (CCK-8) assay and transwell assay. Western blot and enzyme-linked immunosorbent assay (ELISA) were used to verify the effects of hsa_circ_0019217, miR-526b-5p and decorin (DCN) on matrix metalloproteinase 2 (MMP2), tissue inhibitor of metalloproteinase 2 (TIMP2), placental growth factor (PlGF) and human chorionic gonadotropin (hCG) protein expression levels. Dual luciferase reporter gene assay and RNA immunoprecipitation (RIP) assay were used to verify the interaction between hsa_circ_0019217, miR-526-5p and DCN.Results:(1) The analysis of circRNA expression database showed that the expression level of hsa_circ_0019217 was significantly increased in the placental tissues of PE women (fold change=67, P<0.05), and it was mainly located in the cytoplasm. (2) Knockdown of hsa_circ_0019217 promoted the proliferation, migration and invasion of HTR8/SVneo cells, increased the expression levels of MMP2 and PlGF, and decreased the expression levels of TIMP2 and hCG in HTR8/SVneo cells. (3) Hsa_circ_0019217 targeted adsorption of miR-526b-5p, and inhibition of miR-526b-5p reduced the proliferation, migration and invasion of HTR8/SVneo cells. The expression levels of MMP2 and PlGF in HTR8/SVneo cells were increased, and the expression levels of TIMP2 and hCG were decreased. Hsa_circ_0019217 knockdown and inhibiting miR-526b-5p could reverse the effect of hsa_circ_0019217 knockdown on promoting the proliferation, migration and invasion of HTR8/SVneo cells, and reversed the effect of hsa_circ_0019217 knockdown on the protein expression levels of MMP2, PlGF, TIMP2 and hCG. (4) miR-526b-5p targeted DCN in HTR8/SVneo cells, hsa_circ_0019217 knockdown reduced the expression level of DCN, and inhibiting miR-526b-5p increased the expression level of DCN. When hsa_circ_0019217 and miR-526b-5p were inhibited simultaneously, there was no significant change in the protein expression level of DCN. (5) Overexpression of miR-526b-5p promoted the proliferation, migration and invasion of HTR8/SVneo cells, while overexpression of DCN inhibited the proliferation, migration and invasion of HTR8/SVneo cells. Simultaneous overexpression of miR-526b-5p and DCN reversed the effects of miR-526b-5p overexpression on cell proliferation, migration and invasion. Overexpression of miR-526b-5p increased the expression levels of MMP2 and PlGF and decreased the expression levels of TIMP2 and hCG in HTR8/SVneo cells. Overexpression of DCN reduced the expression levels of MMP2 and PlGF and increased the expression levels of TIMP2 and hCG. Overexpression of miR-526b-5p and DCN reversed the effects of miR-526b-5p on the expression of these proteins. Conclusion:Hsa_circ_0019217 regulates the expression of DCN by adsorption of miR-526b-5p, thereby affecting the proliferation, migration and invasion of trophoblast cells and regulating the protein expression levels of MMP2, TIMP2, PlGF and hCG, which might be used as a target for early prevention of PE.

Objective To explore the effect of the blood flow restriction(BFR)at 40%arterial oc-clusive pressure(AOP)on ergometer cycling maximal lactate steady state(MLSS).Methods A total of 11 male college students majoring in sports science(age 23±2 yrs,height 176±5 cm,weight 74.6±5.5 kg,body fat 14.5%±4.7%)were selected.The test in this study was divided into 4 parts:① an incremental ramp test to determine the maximal aerobic power(Pmax);② the MLSS test to determine the blood lactate concentration of MLSS(MLSSc),the work load of MLSS(MLSSw),and the percentage of MLSSw relative to Pmax(%MLSSw);③ the 30 min constant load BFR test(MLSS-BFR)of MLSSw based on the test ② to determine the heart rate,blood lactate and subjec-tive fatigue of MLSSw at the BFR;④ MLSS test at BFR(BFR-MLSS)to determine MLSSc and MLSSw.The BFR was performed using an adjustable pressure compression cuff applied externally to the nearest point to the thigh,at a pressure of 40%AOP.Heart rate was monitored throughout the test.When measuring the constant load in test ②③④,restrictive pressure was released for 30 s ev-ery 5 min.During the release,a blood sample was collected from the earlobe for analysis of blood lac-tate.After the constant load test,the perceived exertion was collected immediately.Results MLSSw(152.5±28.8 vs 161.3±28.1 W,P<0.05,ES=0.84)and%MLSSw(53.4%±5.7%vs 56.7%±5.5%,P<0.05,ES=0.82)of BFR-MLSS test were significantly lower than those of MLSS test.However,no significant differences were found between the BFR-MLSS and MLSS test in MLSSc(5.61±1.18 vs 5.61±0.81 mmol/L,P>0.05,ES=0.01),heart rate(152.6±14.8 vs 150.7±10.7 bpm,P>0.05,ES=0.17)and RPE(14.8±3.3 vs 14.9±2.9,P>0.05,ES=0.06).Conclusion BFR exercise achieves MLSS at a lower external load(power output),and does not reduce the internal load of MLSS.Moreover,BFR increases the internal load for the same external load,but the division of the internal load interval seems to be the same during exercise with or without BFR.

Objectives:To investigate the effect of circular RNA (circRNA) hsa_circ_0019217 on the biological function of chorionic trophoblast cell line (HTR8/SVneo) and its mechanism in the occurrence of pre-eclampsia (PE).Methods:The circRNA expression database was used to analyze the differentially expressed circRNA in placental tissues of PE women. The expression levels of hsa_circ_0019217 and miR-526b-5p were detected by reverse transcription real-time fluorescent quantitative PCR (RT-qPCR), and the subcellular localization of hsa_circ_0019217 was detected by fluorescence in situ hybridization. The proliferation, migration and invasion of trophoblast cells were detected by cell counting kit-8 (CCK-8) assay and transwell assay. Western blot and enzyme-linked immunosorbent assay (ELISA) were used to verify the effects of hsa_circ_0019217, miR-526b-5p and decorin (DCN) on matrix metalloproteinase 2 (MMP2), tissue inhibitor of metalloproteinase 2 (TIMP2), placental growth factor (PlGF) and human chorionic gonadotropin (hCG) protein expression levels. Dual luciferase reporter gene assay and RNA immunoprecipitation (RIP) assay were used to verify the interaction between hsa_circ_0019217, miR-526-5p and DCN.Results:(1) The analysis of circRNA expression database showed that the expression level of hsa_circ_0019217 was significantly increased in the placental tissues of PE women (fold change=67, P<0.05), and it was mainly located in the cytoplasm. (2) Knockdown of hsa_circ_0019217 promoted the proliferation, migration and invasion of HTR8/SVneo cells, increased the expression levels of MMP2 and PlGF, and decreased the expression levels of TIMP2 and hCG in HTR8/SVneo cells. (3) Hsa_circ_0019217 targeted adsorption of miR-526b-5p, and inhibition of miR-526b-5p reduced the proliferation, migration and invasion of HTR8/SVneo cells. The expression levels of MMP2 and PlGF in HTR8/SVneo cells were increased, and the expression levels of TIMP2 and hCG were decreased. Hsa_circ_0019217 knockdown and inhibiting miR-526b-5p could reverse the effect of hsa_circ_0019217 knockdown on promoting the proliferation, migration and invasion of HTR8/SVneo cells, and reversed the effect of hsa_circ_0019217 knockdown on the protein expression levels of MMP2, PlGF, TIMP2 and hCG. (4) miR-526b-5p targeted DCN in HTR8/SVneo cells, hsa_circ_0019217 knockdown reduced the expression level of DCN, and inhibiting miR-526b-5p increased the expression level of DCN. When hsa_circ_0019217 and miR-526b-5p were inhibited simultaneously, there was no significant change in the protein expression level of DCN. (5) Overexpression of miR-526b-5p promoted the proliferation, migration and invasion of HTR8/SVneo cells, while overexpression of DCN inhibited the proliferation, migration and invasion of HTR8/SVneo cells. Simultaneous overexpression of miR-526b-5p and DCN reversed the effects of miR-526b-5p overexpression on cell proliferation, migration and invasion. Overexpression of miR-526b-5p increased the expression levels of MMP2 and PlGF and decreased the expression levels of TIMP2 and hCG in HTR8/SVneo cells. Overexpression of DCN reduced the expression levels of MMP2 and PlGF and increased the expression levels of TIMP2 and hCG. Overexpression of miR-526b-5p and DCN reversed the effects of miR-526b-5p on the expression of these proteins. Conclusion:Hsa_circ_0019217 regulates the expression of DCN by adsorption of miR-526b-5p, thereby affecting the proliferation, migration and invasion of trophoblast cells and regulating the protein expression levels of MMP2, TIMP2, PlGF and hCG, which might be used as a target for early prevention of PE.

Objective:To explore the activation pathway of CD4+CD28-T cells and their specific role in rheumatoid arthritis(RA).Methods:Peripheral blood mononuclear cells(PBMCs)were isolated from RA patients and healthy individuals.Flow cytome-try was used to detect the proportion and activation level of CD4+CD28-T cells,as well as the expressions of perforin and TNF-α,and to analyze the correlation between the proportion of CD4+CD28-T cells and age,C-reactive protein,anti-cyclic citrullinated peptide(CCP)antibodies,rheumatoid factors(RF)and erythrocyte sedimentation rate(ESR).The expression of Toll-like receptor(TLR)2 on CD4+CD28-T cells was detected by flow cytometry.Purified RA CD4+T cells were stimulated with anti-CD3 antibody and TLR2 agonist alone or in combination,and the activation level and cytokine secretion of CD4+CD28-T cells were detected by flow cytometry.Results:The proportion of CD4+CD28-T cells in RA patients was higher than that in healthy individuals(P<0.05),and was positively correlated with rheumatoid factors and erythrocyte sedimentation rate(r=0.554 1,P<0.01;r=0.363 4,P<0.01).Compared with CD4+CD28+T cells,CD25 expression was downregulate(P<0.01),but CD40L and TLR2 expressions were upregulated(P<0.01;P<0.000 1)in CD4+CD28-T cells from RA patients,and they secreted more perforin and TNF-α(P<0.001;P<0.001).After TLR2 ago-nist stimulation,there was no significant change in CD40L and CD25 expression,as well as perforin and TNF-α secretion in CD4+CD28-T cells from RA patients.Conclusion:The proportion of CD4+CD28-T cells is increased in RA patients,and they secrete a large amount of perforin and TNF-α,which may be involved in the inflammatory state and joint damage of RA,but TLR2 cannot re-place CD28 in activating CD4+CD28-T cells.

Objective To explore the effect of the blood flow restriction(BFR)at 40%arterial oc-clusive pressure(AOP)on ergometer cycling maximal lactate steady state(MLSS).Methods A total of 11 male college students majoring in sports science(age 23±2 yrs,height 176±5 cm,weight 74.6±5.5 kg,body fat 14.5%±4.7%)were selected.The test in this study was divided into 4 parts:① an incremental ramp test to determine the maximal aerobic power(Pmax);② the MLSS test to determine the blood lactate concentration of MLSS(MLSSc),the work load of MLSS(MLSSw),and the percentage of MLSSw relative to Pmax(%MLSSw);③ the 30 min constant load BFR test(MLSS-BFR)of MLSSw based on the test ② to determine the heart rate,blood lactate and subjec-tive fatigue of MLSSw at the BFR;④ MLSS test at BFR(BFR-MLSS)to determine MLSSc and MLSSw.The BFR was performed using an adjustable pressure compression cuff applied externally to the nearest point to the thigh,at a pressure of 40%AOP.Heart rate was monitored throughout the test.When measuring the constant load in test ②③④,restrictive pressure was released for 30 s ev-ery 5 min.During the release,a blood sample was collected from the earlobe for analysis of blood lac-tate.After the constant load test,the perceived exertion was collected immediately.Results MLSSw(152.5±28.8 vs 161.3±28.1 W,P<0.05,ES=0.84)and%MLSSw(53.4%±5.7%vs 56.7%±5.5%,P<0.05,ES=0.82)of BFR-MLSS test were significantly lower than those of MLSS test.However,no significant differences were found between the BFR-MLSS and MLSS test in MLSSc(5.61±1.18 vs 5.61±0.81 mmol/L,P>0.05,ES=0.01),heart rate(152.6±14.8 vs 150.7±10.7 bpm,P>0.05,ES=0.17)and RPE(14.8±3.3 vs 14.9±2.9,P>0.05,ES=0.06).Conclusion BFR exercise achieves MLSS at a lower external load(power output),and does not reduce the internal load of MLSS.Moreover,BFR increases the internal load for the same external load,but the division of the internal load interval seems to be the same during exercise with or without BFR.

Objective To develop an enteral nutrition management system for critically ill patients and assess its application outcomes to standardize enteral nutrition management.Methods Based on relevant guidelines and indicator systems,a management system for enteral nutrition in critically ill patients was constructed,consisting of 4 modules:nutritional screening and assessment,nutritional implementation,nutritional monitoring,and statistical analysis.A convenience sampling method was used to select enteral nutrition patients and healthcare staff from the ICU of a tertiary hospital in Ningbo.Data from January to February 2024 served as an experimental group,while data from January to February 2023 constituted a control group.The 2 groups were compared regarding nutritional risk screening rate,feeding interruption rate,completion rate of the enteral nutrition plan,and incidence of complications.At the same time,the system's effectiveness was assessed by healthcare professionals using the clinical nursing information system effectiveness evaluation form.Results The study included 111 patients in the experimental group and 101 patients in the control group.The experimental group exhibited a significantly higher nutritional risk screening rate and enteral nutrition plan completion rate,as well as significantly lower feeding interruption rate and incidence of mechanical complications compared to the control group(P＜0.05).The system received a high effectiveness rating,with an average score of 104.73±9.34.Conclusion The application of the enteral nutrition management system effectively improves the nutritional risk screening rate and completion rate of enteral nutrition plans,while reducing both the feeding interruption rate and the incidence of mechanical complications.Healthcare staff highly rated the system.