OBJECTIVE To investigate the improvement effects and mechanism of astragaloside Ⅳ （AS- Ⅳ ） on lipopolysaccharide （LPS）-induced neuroinflammation. METHODS BV2 cells were divided into control group， LPS group， AS-Ⅳ groups at concentrations of 20 and 40 μmol/L， and dexamethasone group （2 μmol/L）. Except for control group， neuroinflammation model was established with LPS （1 μg/mL） in other groups after medication. The levels of inflammatory factors [interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， and nitric oxide （NO）] in cell supernatant were measured in each group. Mice were randomly divided into normal group， model group， positive control group （Aspirin enteric-coated tablet， 20 mg/kg）， AS-Ⅳ low- and high-dose groups （10， 20 mg/kg）， with 6 mice in each group. Mice in each group were administered the corresponding drug/normal saline via gavage/intraperitoneal injection， once a day， for 14 consecutive days. Except for normal group， other groups were intraperitoneally injected with LPS （250 μg/kg） 1 hour after daily administration of the drug/normal saline to establish neuroinflammation model. Serum levels of IL-6 and TNF-α were measured 2 h after the last medication； histopathological morphology of cerebral tissue in mice were observed； the co-localization of inducible nitric oxide synthase （iNOS）/ionized calcium binding adapter molecule 1 （Iba1） and CD206/Iba1 in the cerebral cortex region of mice was observed； the expressions of proteins related to the nuclear factor-κB （NF-κB）/mitogen-activated protein kinase （MAPK） signaling pathway in brain tissue of mice were also determined， including NF-κB p65， phosphorylated NF-κB p65（p-NF-κB p65）， p38 MAPK， phosphorylated p38 MAPK （p-p38 MAPK）， extracellular signal-regulated kinase （ERK）， and phosphorylated ERK （p-ERK）. RESULTS In the cell experiments， compared with control group， the levels of IL-6， TNF- α and NO in the cell supernatant of the LPS group were increased significantly （P＜0.05）； compared with LPS group， the levels of IL-6， TNF-α and NO were decreased significantly in the administration groups （P＜0.05）. In the animal experiments， compared with the normal group， the serum levels of IL-6 and TNF- α， the number of iNOS/Iba1 co-localization positive cells in the cerebral cortex， and the phosphorylation levels of p38 MAPK， NF- κB p65 and ERK proteins in brain tissue were all significantly increased/elevated in model group （P＜0.05）； the number of CD206/ Iba1 co-localization positive cells in the cerebral cortex region significantly decreased （P＜0.05）. The neurons in the cerebral cortex and the CA3 region of the hippocampus displayed a disordered arrangement. Compared with model group， above quantitative indexes of mice were all reversed significantly in administration groups （P＜0.05）； the neuronal cells in the cerebral cortex and the CA3 region of the hippocampus exhibited a relatively orderly arrangement. CONCLUSIONS AS-Ⅳ may inhibit the activation of the NF-κB/MAPK signaling pathway， promote the M2-type polarization of microglia， and thereby suppress neuroinflammatory responses.

Purpose: This study aimed to evaluate the safety and efficacy of gemcitabine and cisplatin (GP) regimen in combination with immune checkpoint inhibitor sintilimab as neoadjuvant therapy for muscle-invasive bladder cancer (MIBC) patients and the feasibility of the following selective bladder sparing surgery. Materials and Methods: Patients with histopathologically confirmed urothelial carcinoma without distant metastases (T2-4a, N ≤ 1, M0, American Joint Committee of Cancer 8th) and with adequate organ function will be enrolled. The therapeutic regimen was sintilimab 200 mg once on day 8, gemcitabine 1,000 mg/m2 and cisplatin 35 mg/m2 once on days 1 and 8, every 21 days for four cycles. The primary endpoint was pathologic complete response (pCR, pT0N0) rate. The secondary end points were ypT < 2 rate, R0 resection rate, event-free survival, and safety. Results: From May 4, 2020, to May 20, 2023, 55 patients were enrolled. Forty-six patients were evaluated for efficacy. Among the 42 patients who underwent surgery, 16 patients (38.0%) achieved pCR. Thirty-three patients (78.6%) achieved pT < 2. With a median follow-up of 15.7 months, the 1-year event-free survival was 91.3%. Notwithstanding the poor pathological baseline characteristic of a high T3-T4a proportion (39.1%), a promising bladder preservation (including 22 patients transurethral resection of bladder tumor, 5 patients partial cystectomy, and 4 surveillances) rate was achieved (67.4%). The most common grade ≥ 3 treatment-related adverse events was neutropenia (n=15, 27.3%), which was related to chemotherapy. There were no grade 3 immune-related adverse events. Conclusion Neoadjuvant GP plus sintilimab is a promising regimen for MIBC patients, with relatively high pT < 2 rate and triggering the emerging roles for the multi-disciplinary team decision-making for bladder sparing surgery.

Objective:To construct a prognostic model for ovarian cancer based on programmed cell death receptor-1(PD-1)and B lymphocyte infiltration.Methods:A total of 102 patients with ovarian cancer admitted to the First Affiliated Hospital of Gannan Medical College from February 2019 to February 2022 were selected.PD-1 expression and CD20+B cell infiltration degree in cancer tissues were detected,relationship between them and pathological characteristics were analyzed,and correlation between PD-1 expres-sion with B lymphocyte infiltration degree and survival rate of patients were analyzed.Independent influencing factors of poor prognosis of ovarian cancer patients were analyzed,prediction model was built and validated according to independent factors.Results:PD-1 expression and CD20+B cell infiltration degree were closely related to tumor size,FIGO stage,tumor differentiation and lymph node metastasis(P<0.05).PD-1 expression was negatively correlated with B-lymphocyte infiltration degree(P<0.05).Patients with high PD-1 expression and low B-lymphocyte infiltration had higher mortality(P<0.05).FIGO stage,lymph node metastasis,PD-1 expres-sion and CD20+B cell infiltration were independent factors affecting poor prognosis of ovarian cancer patients.Prediction model was constructed by independent influencing factors,which had good differentiation and accuracy.Conclusion:PD-1 expression is closely related to CD20+B cell infiltration degree.There is a close relationship between PD-1 and CD20+B cell infiltration with clinico-pathological characteristics and prognosis of ovarian cancer patients,which are independent influencing factors for poor prognosis of ovarian cancer patients.

Objective To explore the effects and mechanism of calycosin-7-glucoside(CG)on lipopolysaccharide(LPS)-induced inflammatory injury in BV-2 cells and in a mouse model of neuroinflammation.Methods An in vitro neuroinflammation model was induced by LPS stimulation of BV-2 cells.BV-2 cells were divided into blank(CON),model(LPS),dexamethasone(DEX),and low-and high-dose CG(CG 10 μmol/L,CG 20 μmol/L,respectively)groups.The cell viability in each group was detected by Cell Counting Kit-8 assay,interleukin(IL)-6 and tumor necrosis factor(TNF)-α levels in the supernatant were detected by enzyme-linked immunosorbent assay(ELISA),and nitric oxide levels were detected using the Griess method.LPS was also used to induce neuroinflammation in mice in vivo.The mice were then divided randomly into blank(CON),model(LPS),aspirin,and low-and high-dose CG(CG 5 mg/kg,CG 10 mg/kg,respectively)groups.Pathological changes in the hippocampus were detected by hematoxylin/eosin staining.Serum levels of IL-6 and TNF-α were detected by ELISA,polarization of microglia was detected by immunofluorescence staining,and protein expression levels of Toll-like receptor 4(TLR4),myeloid differentiation primary response 88(MyD88),nuclear factor κB(NF-κB,P65)and phosphorylated-NF-κB(p-P65)in the cortex were detected by Western blot.Results CG alone or in combination with LPS in the concentration range of 2.5～160 μmol/L had no significant toxicity in BV-2 cells in vitro,compared with the CON group(P＞0.05).IL-6,TNF-α,and NO levels in the cell supernatant were increased in the LPS group compared with the CON group(P＜0.01),but were significantly reduced by CG(P＜0.05,P＜0.01).Hippocampal neurons were arranged loosely and disordered in the LPS group in vivo,compared with the CON group,and nuclear pyknosis was observed.Serum levels of IL-6 and TNF-α were increased(P＜0.05,P＜0.01).The number of ionized calcium binding adaptor molecule 1(Iba1)/inducible nitric oxide synthase(iNOS)cells was increased(P＜0.01),the number of CD206/Iba1 cells was decreased(P＜0.01),and expression levels of TLR4,MyD88,and p-P65 protein in the cortex were increased(P＜0.05).Compared with the LPS group,CG improved the pathological damage to the hippocampus and inhibited serum levels of IL-6 and TNF-α(P＜0.01).CG also decreased the number of iNOS/Iba1 cells,increased the number of CD206/Iba1 cells(P＜0.05,P＜0.01),and significantly down-regulated TLR4,MyD88,and p-P65 protein levels in the cortex(P＜0.05).Conclusions CG can ameliorate neuroinflammation in mice by suppressing the TLR4/MyD88/NF-κB pathway.

Objective To analyze challenges in translating exercise management evidence for patients with metabolism-associated fatty liver disease(MAFLD),develop actionable strategies,and evaluate the application of best evidence.Methods Utilizing the evidence translation model,the best evidence was implemented for MAFLD patients in 4 phases:evidence acquisition,baseline practice review,intervention,and outcome evaluation.We compared the knowledge of exercise management evidence,implementation rates of review indicators,completion of exercise programs,BMI,liver stiffness measurement,controlled attenuation parameters,and patient satisfaction among medical staff at a tertiary hospital in Fujian Province during baseline(March-May 2023),mid-practice(June-August 2023),and late-practice(September-November 2023)phases.Results A total of 88 patients were included at baseline review,95 during mid-practice,and 107 in late-practice.Significant improvements were observed in the implementation rates of 21 review indicators,nurses'knowledge,completion rate,BMI,and controlled attenuation parameters compared to the data at baseline(P＜0.05).Conclusion The application of best evidence in exercise management for MAFLD patients enhances nurses'knowledge,standardizes nursing practices,and reduces patients'BMI and controlled attenuation parameters.

Objective To study how lipid bilayer fluidity modulates the interaction between β1 integrin and CD40L,as well as the formation of CD40L-mediated tumor cell contact interfaces.Methods Supported lipid bilayers(SLB)with different fluidities were prepared through adjusting the 1,2-dioleoyl-sn-glycero-3-[N-(5-amino-l-carboxypentyl)iminodiacetic acid]succinyl nickel salt(DGS-NTA)content.The functionalization of lipid bilayers was achieved by anchoring fluorescently labeled CD40L molecules onto the membrane surface.The contact interface formation of PC9 cells on the functionalized lipid bilayers was observed through confocal fluorescence imaging and fluorescence recovery after photobleaching(FRAP)experiments,and data of two dimensional(2D)reaction kinetics of β1 integrin and CD40L were extracted from Zhu-Golan plots.Results The diffusion coefficient of molecules in lipid bilayer was negatively correlated with DGS-NTA content.High fluidity of lipid bilayer promoted CD40L accumulation at cell contact interface and expanded the cell contact area.The 2D dissociation constants(2D Kd)of β1 integrin-CD40L complexes were approximately 13,31 and 65 molecules/μm2 for the three lipid bilayers with high,moderate and low fluidities,respectively.Conclusions High fluidity of lipid bilayers significantly facilitates diffusion and aggregation of CD40L to the cell contact interface,thus enhancing β1 integrin-CD40L interaction and the stability of cell contact interfaces.

Objective This study aimed to investigate how β-glucosidase(GBA)knockdown affects malignant progression in cisplatin(DDP)-resistant ovarian cancer(OC)cells and the role of the EGFR signaling pathway.Methods The A2780/DDP cells were categorized into four groups,with one of them serving as blank control(Con)group,si-NC group(transfected with negative control si-NC),si-GBA group(transfected with si-GBA),and NSC 228155 group(transfected with si-GBA and treated with 2 μmol/L NSC 228155).The protein expression levels of GBA,E-cadherin,N-cadherin,Vimentin,EGFR,p38 mitogen-activated protein kinase(p38 MAPK),phospho(p)-p38 MAPK,extracellular regulated protein kinase(ERK)and p-ERK were detected through Western blot.The relative expression of GBA was evaluated through reverse transcription quantitative polymerase chain reaction.The proliferation activity,migration,and invasion potential were evaluated using cell counting kit-8(CCK-8),plate clone formation,cell scratch healing,and Transwell migration assays.Thirty-six nude mice were divided into six groups(six mice per group):blank control(injected with normal saline),blank control+DDP(treated with DDP),negative control(injected with A2780/DDP cell suspension transfected with si-NC),negative control+DDP(injected with A2780/DDP cell suspension transfected with si-NC and treated with DDP),knockdown(injected with A2780/DDP cell suspension transfected with si-GBA),and knockdown+DDP groups(injected with A2780/DDP cell suspension transfected with si-GBA and treated with DDP).The tumor volume and weight of nude mice were evaluated.Results The relative protein and mRNA expression levels of GBA were significantly higher in the A2780/DDP group than in the A2780 group(P＜0.05).Compared with estimates in the Con and si-NC groups,the proliferation activity,number of cloned cells,scratch repair rate,and number of transmembrane cells in the si-GBA group were significantly lower(P＜0.05).The abundance of E-cadherin expression exhibited a notable elevation(P＜0.05),and expression levels of Vimentin,N-cadherin,and EGFR as well as the p-p38 MAPK/p38 MAPK and p-ERK/ERK ratios were significantly decreased(P＜0.05).The proliferation activity,number of cloned cells,scratch repair rate,and the count of transmembrane cells and the expression level of E-cadherin in the NSC 228155 group were markedly higher and lower,respectively,than those in the si-GBA group(P＜0.05),and the expression levels of Vimentin,N-cadherin,and EGFR as well as the ratios of p-p38 MAPK/p38 MAPK and p-ERK/ERK were significantly increased(P＜0.05).In the nude mouse xenograft study,the tumor size and mass in the blank control+DDP group were notably smaller and lighter,respectively,compared to those in the blank control group(P＜0.05).The tumor volume and weight were significantly lower in the negative control+DDP group than in the negative control group(P＜0.05),significantly lower in the knockdown+DDP group than in the knockdown group(P＜0.05),moreover,they were markedly reduced in the knockdown group in comparison to both the blank control and negative control groups(P＜0.05).Compared with those in the blank control+DDP and negative control+DDP groups,the tumor volume and weight in the knockdown+DDP group were significantly reduced(P＜0.05).Conclusions GBA knockdown suppresses the proliferation,migration,and invasion of DDP-resistant OC cells significantly as well as the growth of subcutaneous xenografts derived from A2780/DDP cells in nude mice.These effects may be mediated through the inhibition of the EGFR/MAPK/ERK signaling pathway.

Objective To explore the correlation of neutrophil/lymphocyte ratio(NLR),monocyte/high density lipoprotein cholesterol ratio(MHR)and mild cognitive impairment(MCI)in patients with type 2 diabetes mellitus(T2DM).Methods 159 T2DM patients hospitalized in the Department of Endocrinology,Affiliated Hospital of Xuzhou Medical University from February 2023 to April 2023 were selected.The subjects were divided into normal cognitive function(Con,n=72)group and MCI group(n=87)based on score of Montreal cognitive assessment scale(MoCA).NLR and MHR were calculated and compared between the two groups.Spearman correlation analysis was used to analyze the correlation between NLR,MHR and MoCA score.Logistic regression analysis of the factors influencing the association between T2DM with MCI.Receiver operator characteristic(ROC)curve was used to evaluate the predictive value of NLR and MHR for DM complicated with MCI.Results Compared with Con group,MCI group showed a significant increase in hemoglobin A1c(HbA1c),fasting plasma glucose,NLR,MHR,neutrophil count,monocyte count,serum creatinine,cystatin C,while high density lipoprotein cholesterol lymphocytes,visual space and executive function,naming,attention,language ability,abstract thinking,delayed memory,orientation and MoCA score decreased(P＜0.05).Spearman correlation analysis showed that there was a negative correlation between MHR,NLR and MoCA score,visuospatial and executive function,delayed recall and attention.Logistic regression analysis showed that NLR,MHR,HbA1c were all risk factors for MCI in T2DM.ROC curve analysis showed that the area under the curve of NLR combined with MHR for diagnosing T2DM with MCI was 0.872,with corresponding sensitivity of 81.6%and specificity of 87.5%.Conclusions NLR and MHR are closely related to MCI in T2DM patients.The combination of NLR and MHR have certain efficacy in prediction T2DM with MCI.

Objective To explore the effects and mechanism of calycosin-7-glucoside(CG)on lipopolysaccharide(LPS)-induced inflammatory injury in BV-2 cells and in a mouse model of neuroinflammation.Methods An in vitro neuroinflammation model was induced by LPS stimulation of BV-2 cells.BV-2 cells were divided into blank(CON),model(LPS),dexamethasone(DEX),and low-and high-dose CG(CG 10 μmol/L,CG 20 μmol/L,respectively)groups.The cell viability in each group was detected by Cell Counting Kit-8 assay,interleukin(IL)-6 and tumor necrosis factor(TNF)-α levels in the supernatant were detected by enzyme-linked immunosorbent assay(ELISA),and nitric oxide levels were detected using the Griess method.LPS was also used to induce neuroinflammation in mice in vivo.The mice were then divided randomly into blank(CON),model(LPS),aspirin,and low-and high-dose CG(CG 5 mg/kg,CG 10 mg/kg,respectively)groups.Pathological changes in the hippocampus were detected by hematoxylin/eosin staining.Serum levels of IL-6 and TNF-α were detected by ELISA,polarization of microglia was detected by immunofluorescence staining,and protein expression levels of Toll-like receptor 4(TLR4),myeloid differentiation primary response 88(MyD88),nuclear factor κB(NF-κB,P65)and phosphorylated-NF-κB(p-P65)in the cortex were detected by Western blot.Results CG alone or in combination with LPS in the concentration range of 2.5～160 μmol/L had no significant toxicity in BV-2 cells in vitro,compared with the CON group(P＞0.05).IL-6,TNF-α,and NO levels in the cell supernatant were increased in the LPS group compared with the CON group(P＜0.01),but were significantly reduced by CG(P＜0.05,P＜0.01).Hippocampal neurons were arranged loosely and disordered in the LPS group in vivo,compared with the CON group,and nuclear pyknosis was observed.Serum levels of IL-6 and TNF-α were increased(P＜0.05,P＜0.01).The number of ionized calcium binding adaptor molecule 1(Iba1)/inducible nitric oxide synthase(iNOS)cells was increased(P＜0.01),the number of CD206/Iba1 cells was decreased(P＜0.01),and expression levels of TLR4,MyD88,and p-P65 protein in the cortex were increased(P＜0.05).Compared with the LPS group,CG improved the pathological damage to the hippocampus and inhibited serum levels of IL-6 and TNF-α(P＜0.01).CG also decreased the number of iNOS/Iba1 cells,increased the number of CD206/Iba1 cells(P＜0.05,P＜0.01),and significantly down-regulated TLR4,MyD88,and p-P65 protein levels in the cortex(P＜0.05).Conclusions CG can ameliorate neuroinflammation in mice by suppressing the TLR4/MyD88/NF-κB pathway.

Objective:To construct a multidisciplinary collaborative intelligent prevention and treatment system for orthopedic thrombophilia based on advanced intelligent analysis technology and to evaluate its application effectiveness in the management of orthopedic thrombophilia.Methods:Established a clinical multidisciplinary medical team guided by early warning management and human-computer interaction theory, and builded a multidisciplinary collaborative intelligent prevention and treatment system for orthopedic thrombophilia based on intelligent analysis technology.Adopting a prospective pre-and post control study method, 674 orthopedic inpatients at the Second Affiliated Hospital of Nanchang University from February 2021 to October 2022 were selected as study participants by convenient sampling method and divided into a control group (308 cases) and an observation group (366 cases) according to their admission periods. The control group underwent conventional management methods for thrombophilia. In contrast, the observation group utilized a multidisciplinary collaborative intelligent prevention and treatment system for orthopedic thrombophilia for intelligent management methods. The two groups were compared regarding risk assessment rates, assessment accuracy, assessment efficiency, dynamic assessment completion rate, knowledge awareness rates, implementation rates of multidisciplinary collaborative preventive measures, and thrombophilia incidence rates.Results:The multidisciplinary collaborative intelligent prevention and treatment system for orthopedic thrombophilia mainly included four personnel ports (doctor end, nurse end, patient or caregiver end, medical quality control end) and eight modules (patient intelligent scoring, risk grading warning, dynamic evaluation of prevention contraindications, prevention measure decision-making, health education, inpatient data monitoring, department indicator statistics, medical management). Both groups of patients completed the study. There were 238 males and 70 females in the control group, aged (42.83 ± 8.69) years old. There were 278 males and 88 females in the observation group, aged (42.35 ± 8.13) years old. The risk assessment rate, assessment accuracy rate, and dynamic assessment completion rate of the observation group for thrombophilia were 100.00% (366/366), 98.90% (362/366), and 100.00% (366/366), respectively, all higher than the control group's 94.15% (290/308), 90.58% (279/308), and 91.55% (282/308), and the differences were statistically significant ( χ2=21.99, 24.88, 32.16, all P<0.01). The knowledge awareness rate of preventing thrombophilia among high-risk patients in the observation group was 95.90% (211/220), the implementation rates of basic prevention, physical prevention, drug prevention, and combined prevention in the multidisciplinary collaborative prevention measures were 87.37% (173/198), 97.72% (215/220), 39.09% (86/220), 46.37% (102/220), and 27.73% (61/220), respectively, all higher than the control group's 85.86% (170/198), 24.74% (49/198), 30.81% (61/198), and 12.12% (24/198), and the differences were statistically significant ( χ2 values were 9.81-20.19, all P<0.05). The risk assessment time for the observation group was (3.95 ± 1.03) minutes, and the incidence of thrombophilia was 1.91% (7/366), both lower than the control group's (9.56 ± 1.65) minutes and 7.47% (23/308), with statistically significant differences ( t=53.78, χ2=6.33, both P<0.05). Conclusions:The application of the multidisciplinary collaborative intelligent prevention and treatment system for orthopedic thrombophilia has improved the implementation rate of prevention and treatment interventions by medical staff, increased job satisfaction, and also enhanced the awareness rate of prevention knowledge among patients, thereby improving the management status of thrombophilia in orthopedic inpatients.