The purpose of this study was to investigate the effect of medium-chain fatty acids(MC-FAs)caprylic acid(C8∶0)on lipid metabolism of calf hepatocytes.Primary calf hepatocytes were extracted and cultured,and 1.2 mmol/L nonesterified fatty acids(NEFAs)were added to the hep-atocytes to construct a model of hepatic lipid deposition in primary calf hepatocytes,Five process-ing groups have been set up:Control group(Ctrl),NEFA added group(NEFA),C8∶0 1.2 mmol/L treatment group(C8∶0 1.2),NEFA+C8∶0 0.2 mmol/L treatment group(NEFA+C8∶00.2),C8∶0 0.2 mmol/L treatment group(C8∶0 0.2).Stimulate calf liver cells for 12 hours,and the levels of triglyceride(TG),lipid oxidation(MDA),hydrogen peroxide(H2O2)and total SOD activity were detected by biochemical kit,and FAS,a protein related to lipid synthesis,was detec-ted by Western blot.The results showed that compared with the control group,the concentrations of TG,MDA and H2O2 in NEFA group increased significantly(P＜0.01),and the activity of SOD decreased significantly(P＜0.05).The protein expression levels of FAS,ACC1,DGAT2 and SREBP-1C were significantly up-regulated(P＜0.01),while the expression level of CPT1A was significantly down-regulated(P＜0.01).Compared with the NEFA group,the protein expression levels of SREBP-1C and DGAT2 in the NEFA+C8∶0(concentration 0.2 mmol/L)group de-creased significantly(P＜0.05),and the protein expression level of fatty acid β-oxidation related molecule CPT1A was slightly higher than that in the NEFA group,but there was no statistical sig-nificance(P＞0.05),and the MDA level in hepatocytes decreased significantly(P＜0.05).In a word,the results of this study show that C8∶0 has antioxidant effect,which can effectively reduce the liver injury caused by oxidative stress,regulate the expression of liver fat gene,and then pro-tect liver injury.

The peripheral blood of healthy or ketosis dairy cows was collected,and CD4+T cells were isolated.The expressions of lipid synthesis related proteins fatty acid synthase(FASN),acetyl coenzyme A carboxylase 1(ACC1),cluster of differentiation 36(CD36)and store-operated calcium entry(SOCE)related proteins ORAIl,ORAI2,ORAI3,STIM1,STIM2 were detected by Western blot.IL-17 cells were detected by flow cytometry.CD4+T cells were isolated from the spleen of 1-day-old calves and cultured in vitro.Cells were treated and divided into control(Ctrl)group,si-lenced CD36(siCD36)group,stearic acid(SA)group,and SA+siCD36 group.Cells in the Ctrl and SA groups were transfected with 75 pmol/L negative control siRNA for 48 h,and then stimulated with 200 μmol/L SA for 24 h;Cells in the siCD36 group and SA+siCD36 group were transfected with 75 pmol/L CD36 siRNA for 48 h,and then stimulated with 200 μmol/L SA for 24 h in the SA+siCD36 group.The protein expression of FASN,CD36,ACC1,ORAI1,ORAI2,ORAI3,STIM1 and STIM2 was detected by Western blot,and IL-17 cells were detected by flow cytometry.The results showed that the expression of IL-17 in peripheral blood CD4+T cells of ketosis dairy cows was significantly increased compared to that of healthy cows(P＜0.01).Additionally,the protein level of FASN,CD36,STIM1(P＜0.05),and ACC1,ORAI2,ORAI3,STIM2(P＜0.01)were up-regulated.Compared with the Ctrl group,the protein expression levels of CD36,ACC1 and ORAI3(P＜0.05)were up-regulated in the SA group,as well as the protein expression of FASN and STIM1(P＜0.01).Additionally,the expression of IL-17 was significantly increased(P＜0.05).Compared with the SA group,there was a decrease in the protein expression of STIM1,ORAI1(P＜0.05)and CD36,ACC1,FASN,ORAI2(P＜0.01)in the siCD36+SA group,as well as IL-17(P＜0.05).These results suggest that SA can promote the expression of IL-17 in CD4+T cells in ketosis cows by regulating fatty acid synthesis and activating SOCE channels through CD36.

Objective To investigate the effect and potential mechanism of rat mesenchymal stem cells(MSC)on D-galactose-induced brain-tissue aging.Methods A rat brain-aging model was established by injecting D-galactose,and rats in the treatment group received MSC injections via the tail vein.Superoxide dismutase(SOD)activity and malondialdehyde(MDA)levels were assessed in rat brain tissue at the end of the experiment,and pathological changes in brain tissue were observed by hematoxylin-eosin(HE)staining.Expression levels of the inflammatory factors interleukin(IL)-1 and IL-6,the pathway proteins brain-derived neurotrophic factor(BDNF)-tropomyosin receptor kinase B(TrkB),the negative growth regulators p53 and p16,as well as vascular endothelial growth factor(VEGF)and basic fibroblast growth factor(bFGF)were observed by polymerase chain reaction(PCR)and Western Blot.Results Brain levels of SOD activity were significantly increased and MDA levels were significantly decreased in rats in the modle group compared with the treatment group(P＜0.05).The pathological state of the cerebral cortex and hippocampus were improved and the number of neurons and nucleus pulposus ratio in the brain were increased in the treatment group,as shown by HE staining.Expression levels of IL-1,IL-6,p53,and p16 were significantly decreased,while BDNF,TrkB,VEGF,and bFGF were significantly increased in the treatment group compared with the model group,as shown by PCR and Western Blot(P＜0.05).Conclusions These result suggest that MSCs potentially mitigate D-galactose-induced cerebral senescence by concurrently modulating the BDNF-TrkB axis to attenuate oxidative/inflammatory damage,while enhancing the secretion of vasculotrophic(VEGF)and neurotrophic(bFGF)factors for neuronal maintenance.

Purpose To explore the pathological characteristics,diagnosis,and differential diagnosis of pure ery-throid leukemia(PEL).Methods A retrospective analysis was conducted on the clinicopathological data of 12 cases of PEL.Immunohistochemical EnVision method and flow cytometry were used to detect PEL-related immune markers,and heat-treated Giemsa R-banding technique was applied to analyze the chromosomal karyotype.Results Peripheral blood and bone marrow smears revealed that 2 out of 7 cases showed presence of proerythroblast in peripheral blood,and 7 out of 12 cases showed atypical proerythroblast in bone marrow samples.After recounting,the average percentage of proerythroblast in the 12 PEL cases was 36.8％(ranging from 2％to 69.5％),with an average of 53.2％of all er-ythroid cells(ranging from 5％to 88％).Among them,9 cases did not meet the diagnostic criteria for PEL.Bone marrow biopsy:11 cases showed hypercellularity,with tumor cells showing diffuse proliferation in 9 cases,accompa-nied by dysplasia of megakaryocytes in 7 cases,and there was increased proliferation of fibrous tissue in 9 cases.Im-munohistochemistry:12 cases exhibited strong staining intensity for CD71 and E-cadherin.11 cases expressed CD117,while 4 cases expressed CD34,3 cases exhibited slight expression of GPA,and 1 case weakly expressed CD61.Flow cytometry:in 8 cases,there was an increased proportion of early-stage erythroid cells,accounting for 3.1％to 80.31％of nucleated cells,with an average of 31.0％.All cases expressed CD117 and CD71 to varying degrees,with 7 out of 8 cases expressing CD36,5 out of 7 cases expressing CD105,and 3 out of 4 cases expressing GPA.A few ca-ses demonstrated aberrant expression of CD123 and CD7.Chromosomal Karyotyping:7 cases exhibited highly complex karyotypes(7/8),with frequent involvement of chromosomes 5,7,8,17,and 19.One case had a normal karyotype.Conclusion The diagnosis of PEL requires a comprehensive assessment combining various methods including bone marrow smears,bone marrow biopsy,immunohistochemistry,and flow cytometry.

Gastric cancer (GC) is one of the most common malignant tumors of the digestive system in China. With the progress of immunotherapy research, programmed death receptor-1 (PD-1) inhibitor-based combinatory therapy offers new ideas for the treatment of advanced gastric cancer. In recent years, with the increasing status of immunotherapy in the treatment of advanced gastric cancer, a growing number of domestic and international clinical studies shown that immunotherapy could achieve better efficacy in the neoadjuvant therapy and conversion therapy for patients with advanced gastric cancer. This paper reviews the current research progress on the application of PD-1 inhibitors in the neoadjuvant therapy and conversion therapy of gastric cancer.

Objective To construct a dynamic prediction model for the risk of developing Alzheimer's disease(AD)based on baseline clinical data and longitudinal neuropsychological scores in patients with mild cognitive impairment(MCI).Methods A total of 380 MCI patients from the Alzheimer's Disease Neuroimaging Initiative 1 study from 2005 to 2011 were selected and were randomly divided into a training set and a test set in a 7∶3 randomization.The Alzheimer's disease assessment scale-cognitive 13 items(ADAS-Cog13),Rey auditory verbal learning test immediate score(RAVLT Immediate),functional activities questionnaire(FAQ),and mini-mental state examination(MMSE)were used as longitudinal neuropsychological score metrics.Random survival forests with multivariate longitudinal endogenous covariates were used to construct a dynamic prediction model of the risk of developing AD in patients with MCI in the training set.The predictive performance of the model was evaluated in the test set using time-dependent areas under the receiver operator characteristic curve(AUC)and Brier score(BS).Results For the prediction of the risk of developing AD in patients with MCI,longitudinal neuropsychological scores were more important predictors than baseline clinical data,with FAQ being the strongest predictor.The dynamic prediction model had high predictive performance in the test set,with AUC ranging from 0.7695 to 0.8987 and BS ranging from 0.1369 to 0.2184.Conclusion Random survival forests with multivariate longitudinal endogenous covariates can be used to combine multivariate longitudinal neuropsychological scores to construct a prediction model for the risk of developing AD in patients with MCI,with high predictive performance and the ability to achieve individual dynamic prediction.

Objective:To evaluate the efficacy and safety of Rituximab (RTX) combined with short-term use of glucocorticoids in the treatment of the initial episode of steroid-sensitive nephrotic syndrome (SSNS).Methods:A retrospective case summary.A total of 30 children with SSNS treated in the Department of Intrarenal Rheumatism and Immunology, Xuzhou Children′s Hospital from December 2021 to March 2023, were enrolled in this study.They were given a standard dose of RTX (375 mg/m 2) and glucocorticoids for a short term.The patients were followed up for 1 year, and the general condition, changes in CD19 + B lymphocytes expression after RTX treatment, the relapse rate, the median relapse-free survival and adverse reactions of RTX were recorded.Kaplan-Meier method was used to analyze the relapse-free survival time. Results:CD19 + B lymphocytes were depleted 2 weeks after RTX treatment, and the median time required for CD19 + B lymphocytes reconstitution was 5 months after RTX treatment.The 1-year relapse-free rate was 90.00%, and the median relapse-free survival was 11 months.The glucocorticoid discontinuation rate was 93.33% in 3 months after RTX treatment, and 100% in 6 and 12 months after RTX treatment.Adverse reactions included infusion reactions in 7 cases (23.33%), neutropenia/leukopenia in 5 cases (16.67%), hypoimmunoglobulinemia in 10 cases (33.33%), infections in 4 cases.One case was complicated with severe influenza A virus infection after CD19 + B lymphocytes reconstruction. Conclusions:A standard dose of RTX can significantly reduce the relapse frequency, maintain long-term remission of proteinuria, and reduce the dose of glucocorticoids in patients at the initial episode of SSNS.Thus, it is worthy of clinical promotion.

Objective To investigate the effect and potential mechanism of rat mesenchymal stem cells(MSC)on D-galactose-induced brain-tissue aging.Methods A rat brain-aging model was established by injecting D-galactose,and rats in the treatment group received MSC injections via the tail vein.Superoxide dismutase(SOD)activity and malondialdehyde(MDA)levels were assessed in rat brain tissue at the end of the experiment,and pathological changes in brain tissue were observed by hematoxylin-eosin(HE)staining.Expression levels of the inflammatory factors interleukin(IL)-1 and IL-6,the pathway proteins brain-derived neurotrophic factor(BDNF)-tropomyosin receptor kinase B(TrkB),the negative growth regulators p53 and p16,as well as vascular endothelial growth factor(VEGF)and basic fibroblast growth factor(bFGF)were observed by polymerase chain reaction(PCR)and Western Blot.Results Brain levels of SOD activity were significantly increased and MDA levels were significantly decreased in rats in the modle group compared with the treatment group(P＜0.05).The pathological state of the cerebral cortex and hippocampus were improved and the number of neurons and nucleus pulposus ratio in the brain were increased in the treatment group,as shown by HE staining.Expression levels of IL-1,IL-6,p53,and p16 were significantly decreased,while BDNF,TrkB,VEGF,and bFGF were significantly increased in the treatment group compared with the model group,as shown by PCR and Western Blot(P＜0.05).Conclusions These result suggest that MSCs potentially mitigate D-galactose-induced cerebral senescence by concurrently modulating the BDNF-TrkB axis to attenuate oxidative/inflammatory damage,while enhancing the secretion of vasculotrophic(VEGF)and neurotrophic(bFGF)factors for neuronal maintenance.

Objective To analyze the drug resistance and genomic characteristics of clinically isolated hypermuco-viscous Klebsiella pneumoniae(hmKp).Methods Klebsiella pneumoniae isolated from clinical specimens from the sentinel hospitals of National Pathogen Identification Network in Huai'an City from 2019 to 2023 were collected,strains were identified by VITEK 2 Compact,mucus phenotype was determined by string test,resistance of hmKp was determined by microbroth dilution method.Molecular typing was conducted by whole-genome sequencing tech-nology,annotation of virulence and resistance genes carried by strains was performed.Results A total of 60 strains of hmKp were collected,mainly isolated from sputum specimens(33.33％)and blood specimens(28.33％).The average genome size of 60 strains was 5.6 Mb,with an average GC content of 57.09％.Multilocus sequence typing(MLST)showed that there were 28 ST types,with the dominant ST types being ST1 1(11.67％),ST15(11.67％),and ST412(10.00％).Core genome MLST(cgMLST)analysis revealed that some strains were highly homologous,and no outbreak strains were found.Multidrug-resistant strains accounted for 60.00％,while imipe-nem-resistant strains accounted for 28.33％.64 types of resistance genes were carried,84.38％of which were loca-ted on mobile element.The carriage rates of fosfomycin-related resistance genes and extended-spectrum β-lactam an-tibiotic-related resistance genes were relatively high in the strains,at 100％and 98.33％,respectively.Among car-bapenem-resistance gene blaKPC-2,21.67％was carried by ST11,ST1,and ST15 strains,among bla NDM-5 gene,3.33％was carried by ST76 strain.A total of 101 virulence genes were carried,most were Colibactin and type Ⅵsecretion system-related virulence genes.All strains carried capsule synthesis regulation-related genes(rcsA,rcsB),efflux pump-related genes(acrA,acrB),and enterobacterin-related genes(entABCEF,fepABCDFG,fes).Conclusion Clinically isolated hmKp in Huai'an exhibits multidrug resistance,with dominant ST types ST11,ST15,and ST412,carrying multiple horizontally transferable resistance and virulence genes,prevention and control measures should be strengthened.

Purpose To explore the pathological characteristics,diagnosis,and differential diagnosis of pure ery-throid leukemia(PEL).Methods A retrospective analysis was conducted on the clinicopathological data of 12 cases of PEL.Immunohistochemical EnVision method and flow cytometry were used to detect PEL-related immune markers,and heat-treated Giemsa R-banding technique was applied to analyze the chromosomal karyotype.Results Peripheral blood and bone marrow smears revealed that 2 out of 7 cases showed presence of proerythroblast in peripheral blood,and 7 out of 12 cases showed atypical proerythroblast in bone marrow samples.After recounting,the average percentage of proerythroblast in the 12 PEL cases was 36.8％(ranging from 2％to 69.5％),with an average of 53.2％of all er-ythroid cells(ranging from 5％to 88％).Among them,9 cases did not meet the diagnostic criteria for PEL.Bone marrow biopsy:11 cases showed hypercellularity,with tumor cells showing diffuse proliferation in 9 cases,accompa-nied by dysplasia of megakaryocytes in 7 cases,and there was increased proliferation of fibrous tissue in 9 cases.Im-munohistochemistry:12 cases exhibited strong staining intensity for CD71 and E-cadherin.11 cases expressed CD117,while 4 cases expressed CD34,3 cases exhibited slight expression of GPA,and 1 case weakly expressed CD61.Flow cytometry:in 8 cases,there was an increased proportion of early-stage erythroid cells,accounting for 3.1％to 80.31％of nucleated cells,with an average of 31.0％.All cases expressed CD117 and CD71 to varying degrees,with 7 out of 8 cases expressing CD36,5 out of 7 cases expressing CD105,and 3 out of 4 cases expressing GPA.A few ca-ses demonstrated aberrant expression of CD123 and CD7.Chromosomal Karyotyping:7 cases exhibited highly complex karyotypes(7/8),with frequent involvement of chromosomes 5,7,8,17,and 19.One case had a normal karyotype.Conclusion The diagnosis of PEL requires a comprehensive assessment combining various methods including bone marrow smears,bone marrow biopsy,immunohistochemistry,and flow cytometry.