Objective To explore the risk factors of cardiomyopathy in patients with diabetes and their correlations with the levels of serum fork head box protein O1(FOXO1),growth differentiation factor 11(GDF11),and matrix metalloproteinase-3(MMP3).Methods 200 patients with diabetes complicated with cardiomyopathy and 200 patients with simple diabetes admitted to the People's Hospital of Xinjiang Uygur Autonomous Region from September 2023 to April 2025 were selected to detect serum FOXO1,GDF11 and MMP3 levels,analyze their correlation with cardiac function parameters,screen risk factors through multifactor logistic regression,construct a nomogram model and a receiver operating characteristic(ROC)curve,and evaluate the diagnostic value.Results The duration of diabetes cardiomyopathy group≥10 years,the expression rate of reactive oxygen species(ROS)≥90%,the insulin resistance index and the levels of FOXO1 and MMP3 were significantly higher than those in the simple diabetes group(P<0.05),and the level of GDF11 was lower than that in the simple diabetes group(P<0.05).FOXO1 and MMP3 were positively correlated with E/e'and negatively correlated with LVEF and E/A;GDF11 was negatively correlated with E/e'and positively correlated with LVEF and E/A(P<0.05).The nomogram model showed that the weights of the influencing factors were GDF11,FOXO1,insulin resistance index,MMP3,ROS expression rate,and the course of diabetes.The model calibration was good(χ2=7.336,P=0.719).The combined diagnostic AUC of FOXO1,GDF11,and MMP3 was 0.950,which was better than that of a single indicator(P<0.05).Conclusion GDF11,FOXO1,Insulin resistance index,MMP3,ROS expression rate and the course of diabetes are risk factors for diabetes cardiomyopathy,and the combination of the three is of high diagnostic value.

Objective To investigate the functional changes of dendritic cells(DCs)in patients at different stages of hepatitis B virus(HBV)infection and analyze the mechanisms underlying DC dysfunction.Methods Single-cell RNA sequencing dataset GSE182159 was downloaded from the GEO database and classified into healthy control(HC),immune active(IA),and immune tolerant(IT)groups based on infection stage.Peripheral blood samples were collected from 7 IA patients,7 IT patients,and 12 healthy controls.Flow cytometry was used to isolate classical dendritic cells(cDC)and plasmacytoid dendritic cells(pDC).The expression levels of transcription factors in cDC and pDC were measured by quantitative real-time PCR(qRT-PCR).Bioinformatics analyses were per-formed using R and Python package.Results The proportions of DCs in IA and IT groups were higher than that in HC group.Func-tional enrichment analysis revealed that the differentially expressed genes(DEGs)of cDCs in the IA group were primarily enriched for the processes,such as inflammatory response,MHC classⅡantigen processing and presentation,cell migration,signal transduction,metabolism,and immune response.In contrast the IT group exhibited lower enrichment intensity and a significant reduction in interfer-on responses.The DEGs of pDC in the IA group were enriched in the processes of MHC-Ⅱantigen presentation,Fc receptor signal transduction,and metabolism,whereas those in the IT group were showed enrichment only in Fc receptor signal transduction and me-tabolism with a lower intensity.Both groups exhibited reduced synthesis of typesⅠandⅡinterferons in pDC,with the IT group showing a more pronounced downregulation.Cell-cell communication analysis demonstrated enhanced interactions between myeloid cells(except pDC)and T cells in the IA group,whereas the interactions between cDC/pDC and T cells in the IT group were reduced.Transcription factor analysis revealed that STAT2,STAT3,IRF1,and IRF5 were highly expressed in the IA group but their expression exhibited low-er expression levels in the IT group.In contrast,BHLHE40 was broadly upregulated in both cDC and pDC subsets within the IT group.The qRT-PCR results were consistent with the findings from the single-cell transcription factor analysis.Conclusion The IT phase of hepatitis B infection represents a critical period for cDC dysfunction,characterized by significant suppression of MHCⅡantigen presen-tation,metabolism,and interferon responsiveness.The functional impairment of pDC precedes that of cDC,as evidenced by a marked downregulation of interferon synthesis capacity observed during the IA phase.

Objective To investigate the functional changes of dendritic cells(DCs)in patients at different stages of hepatitis B virus(HBV)infection and analyze the mechanisms underlying DC dysfunction.Methods Single-cell RNA sequencing dataset GSE182159 was downloaded from the GEO database and classified into healthy control(HC),immune active(IA),and immune tolerant(IT)groups based on infection stage.Peripheral blood samples were collected from 7 IA patients,7 IT patients,and 12 healthy controls.Flow cytometry was used to isolate classical dendritic cells(cDC)and plasmacytoid dendritic cells(pDC).The expression levels of transcription factors in cDC and pDC were measured by quantitative real-time PCR(qRT-PCR).Bioinformatics analyses were per-formed using R and Python package.Results The proportions of DCs in IA and IT groups were higher than that in HC group.Func-tional enrichment analysis revealed that the differentially expressed genes(DEGs)of cDCs in the IA group were primarily enriched for the processes,such as inflammatory response,MHC classⅡantigen processing and presentation,cell migration,signal transduction,metabolism,and immune response.In contrast the IT group exhibited lower enrichment intensity and a significant reduction in interfer-on responses.The DEGs of pDC in the IA group were enriched in the processes of MHC-Ⅱantigen presentation,Fc receptor signal transduction,and metabolism,whereas those in the IT group were showed enrichment only in Fc receptor signal transduction and me-tabolism with a lower intensity.Both groups exhibited reduced synthesis of typesⅠandⅡinterferons in pDC,with the IT group showing a more pronounced downregulation.Cell-cell communication analysis demonstrated enhanced interactions between myeloid cells(except pDC)and T cells in the IA group,whereas the interactions between cDC/pDC and T cells in the IT group were reduced.Transcription factor analysis revealed that STAT2,STAT3,IRF1,and IRF5 were highly expressed in the IA group but their expression exhibited low-er expression levels in the IT group.In contrast,BHLHE40 was broadly upregulated in both cDC and pDC subsets within the IT group.The qRT-PCR results were consistent with the findings from the single-cell transcription factor analysis.Conclusion The IT phase of hepatitis B infection represents a critical period for cDC dysfunction,characterized by significant suppression of MHCⅡantigen presen-tation,metabolism,and interferon responsiveness.The functional impairment of pDC precedes that of cDC,as evidenced by a marked downregulation of interferon synthesis capacity observed during the IA phase.

AIM:To investigate the effects and mechanism of long noncoding RNA PCED1B antisense strand 1(lncRNA PCED1B-AS1)on the proliferation,migration,invasion and apoptosis of papillary thyroid carcinoma(PTC)cells.METHODS:Human PTC cells were cultured in vitro.The expression of PCED1B-AS1 and fused in sarcoma(FUS)was measured by RT-qPCR.The effects of knockdown/overexpression of PCED1B-AS1/FUS on the migration and invasion of PTC cells were detected via Transwell assay.The effects of knockdown/overexpression of PCED1B-AS1/FUS on PTC cell proliferation were analysed via CCK-8 and plate colony assay.The effect of knockdown PCED1B-AS1 on PTC cell apoptosis was determined by flow cytometry.The target binding of PCED1B-AS1 and FUS was determined with bioin-formatics and RNA immunoprecipitation(RIP)experiments.Fluorescence in situ hybridization experiment was performed to verify whether PCED1B-AS1 colocalises with FUS.The mitogen-activated protein kinase(MAPK)signaling pathway-re-lated proteins were detected via Western blot.RESULTS:(1)PCED1B-AS1 expression was significantly higher and FUS expression was significantly lower in PTC cells compared with normal thyroid Nthy-ori3-1 cell(P<0.05).(2)Knockdown of PCED1B-AS1 and overexpression of FUS inhibited PTC cell migration,invasion and proliferation,and promoted apopto-sis(P<0.05).(3)Bioinformatics analysis and RIP assay verified the existence of targeted binding of PCED1B-AS1 to FUS(P<0.05).(4)PCED1B-AS1 and FUS colocalised in the cytoplasm.(5)Inhibition of PCED1B-AS1 decreased the expression of MAPK signaling pathway-related proteins p-ERK 1/2,p-JNK and p-P38(P<0.05).CONCLUSION:ln-cRNA PCED1B-AS1 inhibits the proliferation,migration and invasion,and promotes the apoptosis of PTC cells,and its mechanism may be related to the expression of FUS and the MAPK signaling pathway.

Objective To investigate the effect of long intergenic non-coding RNA COX2 (lincRNA-COX2) on apoptosis and polarization of Listeria monocytogenes (Lm)-infected RAW264.7 cells. Methods RAW264.7 cells were cultured and divided into control group (uninfected cells), Lm infection group, negative control of small interfering RNA (si-NC) group, si-NC and Lm infection group, small interfering RNA of lincRNA-COX2 (si-lincRNA-COX2) group, si-lincRNA-COX2 and Lm infection group. RAW264.7 cells were infected with MOI=10 Lm for 6 hours, and then the inhibition efficiency of siRNA transfection was detected by fluorescence microscope and quantitative real-time PCR (qRT-PCR). The expression levels of cleaved-caspase-3(c-caspase-3), caspase-3, B-cell lymphoma-2 (Bcl2), Bcl2 associated X protein (BAX), arginase 1 (Arg1), inducible nitric oxide synthase (iNOS) were detected by Western blot analysis. Results c-caspase-3/caspase-3, BAX/Bcl2 and iNOS were significantly up-regulated, while the level of Arg1 was down-regulated in Lm-infected RAW264.7 cells compared with control group. LincRNA-COX2 knockdown inhibited the increase of protein levels for BAX/Bcl2, c-caspase-3/caspase-3 and iNOS in Lm-infected RAW264.7 cells, while the level of Arg1 in Lm-infected RAW264.7 cells was up-regulated. Conclusion Knockdown of lincRNA-COX2 can inhibit cell apoptosis and suppress the macrophage polarization into M1 type in Lm-infected RAW264.7 cells.
Apoptosis/genetics* ; bcl-2-Associated X Protein/metabolism* ; Caspase 3/metabolism* ; Cyclooxygenase 2/metabolism* ; Listeria monocytogenes/pathogenicity* ; Macrophages/microbiology* ; RNA, Long Noncoding/metabolism* ; RNA, Small Interfering/genetics* ; Animals ; Mice

Objective To investigate the effect of Trichomonas vaginalis macrophage migration inhibitory factor (TvMIF) on THP-1 macrophages.. Methods Recombinant TvMIF protein was prokaryotic expressed and purified, and endotoxin was removed after identification. Following exposure to TvMIF at concentrations of 0, 1, 5, 10, 50 and 100 ng/mL, the cytotoxicity of the recombinant TvMIF protein to THP-1 macrophages was tested using cell counting kit (CCK)-8 assay, and the apoptosis of THP-1 macrophages and reactive oxygen species (ROS) were detected using flow cytometry. The relative expression of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), caspase-1, interleukin-1β (IL-1β) and IL-18 genes was quantified using real-time fluorescent quantitative PCR (qPCR) assay, and the expression of caspase-1, NLRP3, gasdermin D (GSDMD), gasdermin D N-terminal (GSDMD-NT) and pro-IL-1β proteins were determined using Western blotting assay. Results Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) displayed successful expression and purification of the recombinant TvMIF protein with a molecular weight of 15.5 kDa, and the endotoxin activity assay showed the successful removal of endotoxin in the recombinant TvMIF protein (endotoxin concentration < 0.1 EU/mL), which was feasible for the subsequent studies on protein functions. Flow cytometry revealed that the recombinant TvMIF protein at a concentration of 10 ng/mL and less promoted the apoptosis of THP-1 macrophages, and the highest apoptotic rate of THP-1 macrophages was seen following exposure to the recombinant TvMIF protein at a concentration of 5 ng/mL, while the recombinant TvMIF protein at concentrations of 50 and100 ng/mL inhibited the apoptosis of THP-1 macrophages. Exposure to the recombinant TvMIF protein at a concentration 1 ng/mL resulted in increased ROS levels in THP-1 macrophages. qPCR assay quantified significantly elevated caspase-1, NLRP3, IL-18 and IL-1β expression in THP-1 macrophages 8 hours post-treatment with the recombinant TvMIF protein at a concentration 1 ng/mL, and Western blotting determined increased caspase-1, NLRP3, pro-IL-1β, GSDMD and GSDMD-NT protein expression in THP-1 macrophages following exposure to the recombinant TvMIF protein at a concentration 1 ng/mL. Pretreatment with MCC950 significantly reduced GSDMD and GSDMD-NT protein expression. Conclusions High-concentration recombinant TvMIF protein inhibits macrophage apoptosis, while low-concentration recombinant TvMIF protein activates NLRP3 inflammasome and promotes macrophage pyroptosis.

Objective To clone and express the thioredoxin(Trx)from RH strain tachyzoites of Toxoplasma gondii,estab?lish the prokaryotic expression vector and purify the recombinant protein,then produce the polyclonal anti?Trx antibody in rab?bits. Methods Trx fragment was amplified by PCR and cloned into the pET?28a(+)vector,and the recombinant protein was in?duced with IPTG and purified by Ni?NTA affinity chromatography. The polyclonal antibody specificity was detected by Western blotting. Results The trx gene was amplified from T. gondii cDNA by PCR. The recombinant plasmid trx/pET?28a(+)was use?fully constructed,and the recombinant TRX protein was expressed and purified. The TRX polyclonal antibody was also ob?tained. The specific band of TRX was detected by Western blotting. Conclusion Western blotting can detect the specificity of polyclonal anti?Trx antibody,which will facilitate the biological functions of Trx.

ObjectiveTo explore the effect of empathy nursing on depressive emotion and life function in depressive patients and to improve treatment and nurse effect.MethodSixties depressive inpatients treated with antidepressive drugs were randomly assigned to receive general nursing and health education ( experimental group 31 ) and empathy nursing (control group 29).Hamilton Depression Scale (HAMD),Nurses'observation scale for inpatient evaluation (NOSIE) and the hospital nursing work satisfaction questionnaire were used before and after intervention to evaluate the clinical and nursing effects of two groups.ResultsAfter interventions,the total score of HAMD was ( 16.7 ± 3.2) in experimental group and (20.2 ± 3.6) in control group,with significant difference between two groups.The social ability factor of score of NOSIE was (9.78 ± 2.13 ) in experimental group and ( 13.32 ± 3.22) in control group,and the scores of 3 other factors of NOSIE in experimental group was obviously lower than that in control group.The score ( 121.37 ± 11.25 ) of nursing work satisfaction in experimental group was obviously higher than that( 113.26 ± 9.72) in control group ( t =2.98).ConclusionsEmpathy nursing can improve depressive emotion and life function of patients with depression,and improve nursing work satisfaction.

Objectives To establish a reliable method for isolating and purifying soluble collagen type Ⅱ(SCⅡ)by optimizing preparative procedure.Method The chicken sternal cartilage was selected as raw material.Guanidine hydrochloride was used to remove the proteoglycans.The digestion manners of pepsin,sodium chloride concentrations for salting,types of DEAE anion resin were studied for extracting SCⅡ.The SCⅡ identification was made by SDS-PAGE,absorption spectrum and amino acid analysis.Result It was convenient for pre-treatments of chicken sternal cartilage.The proteoglycans could be efficiently removed by 4 mol/L guanidine hydrochloride.The satisfied results were obtained by limited enzyme digestion of pepsin added by two steps.The optimizing concentration of sodium chloride for salting was 2.4mol/L.SDS-PAGE maps revealed that the bands of purified SCⅡand standard CⅡ were at the same location.The absorption peak of SCⅡ was at 230nm.The concentrations of Gly,Pro and Ala were the highest in 15 amino acids.Conclusion The improved method has significant advantages of simple working process,result reliability and convenient source of raw material.It is suitable for purifying the SCⅡ at variable scales in research works and clinic application.The SCⅡ product obtained has high purity and accords with the characteristics of collagen type Ⅱ.

Objective To make staining preparation of tapeworm for the purpose of teaching and scientific research.Methoeds Pregnant segmens were stained by Indian ink,and mature segments,cephalomeres and bladder worms were stained by carmine.Result After staining the characteristics of pregnant segments,mature segments,cephalomeres and bladder worms were observed obviously.For instance,the stem and branches linking both-side of uteri in pregnant segment and reproduction cavity projecting in one of edges in the center of the pregnant segment were clearly shown in ink colour after stained by Indian ink.The red colour was shown in the mature segment with deep-coloured male and female reproduction systems inside,separately,after stained by carmine.Conclusion Tapeworm preparation stained by Indian ink and carmine is useful and can be preserved permanently.