Objective: To determine the expression of melanoma-associated antigens D4(MAGED4) and SIRT7 in human glioma, and to analyze the potential effects of MAGED4 and SIRT7 on glioma cell proliferation. Methods: The MAGED4 and SIRT7 expression levels and their correlation were compared by the China glioma genome atlas(CGGA), human protein atlas(HPA), and UALCAN databases. Survival analysis, ROC curve analysis, and Cox regression analysis were used to predict the outcome of MAGED4 and SIRT 7 in glioma patients. Gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG) signaling pathway enrichment analysis were used to explore the biological functions of MAGED4 and SIRT7 in glioma. Western blot experiment was used to investigate whether MAGED4 protein exerted its regulatory effects on the activity of the PI3K/AKT signaling pathway via SIRT7. The effect of MAGED4 on cell proliferation in glioma through SIRT7 was explored by CCK-8. Results: The analysis results of CGGA, UALCAN, and HPA databases showed that the expression levels of MAGED4 and SIRT7 in glioma tissues were higher than those in normal brain tissue, and the expression were positively correlated. Results of survival, ROC, and Cox analysis showed that high expression of MAGED4 and SIRT7 mRNA were risk factors for poor prognosis in glioma. Results of KEGG enrichment analysis showed that MAGED4 and SIRT7 were associated with the PI3K/AKT signaling in glioma, and Western blot results showed that MAGED4 activated the PI3K/AKT signaling pathway by regulating SIRT7. The CCK-8 results showed that MAGED4 promotes the proliferation of glioma cells through SIRT7. Conclusion MAGED4 and SIRT7 are highly expressed in glioma and associated with poor prognosis, and MAGED4 promotes glioma cell proliferation through activation of the PI3K/AKT signaling pathway by SIRT7.

Objective:To evaluate the influencing factors and predictive value of renal function recovery in elderly patients with heart failure(HF)and acute renal injury(AKI)after intermittent veno-venous hemofiltration(IVVH)using critical care ultrasound.Methods:The clinical data of elderly patients with heart failure(NYHF grade Ⅲ~Ⅳ)complicated with acute kidney injury(stage 2~stage 3)who underwent intravenous veno-venous hemofiltration(IVVH)in the critical care unit(CCU)of our hospital were retrospectively analyzed.The demographic information of the patients and the changes in clinical biochemical and critical care ultrasound monitoring parameters before and after 7 days of IVVH were recorded.Based on the recovery of renal function, the patients were divided into two groups: a renal function recovery group and a renal function non-recovery group.Logistic regression and Receiver Operating Characteristic Curve(ROC)curve analysis were performed to determine the predictive value of various influencing factors on the recovery of renal function in patients.Results:A total of 178 patients were enrolled in this study.After starting IVVH treatment, renal function recovered in 143 cases at 30 days, and in 138 cases at 90 days.However, renal function did not recover in 35 cases at 30 days, and in 40 cases at 90 days.The proportion of NYHF Ⅲ patients、the proportion of diabetic patients、the decrease of Beta 2-microglobulin(β2-MC)、the decrease of Cystain C(CysC)、the increase of venous transit time index(VTI)、the increase of Cardiac Output(CO)and the decrease in renal blood flow resistance index(RI)in the recovery groups at both 30 days and 90 days was significantly higher than that in the non-recovery group(all P<0.05).The total treatment time of IVVH in the recovery group was significantly shorter than that in the non-recovery group, with 30 days and 90 days( P<0.05).Logistic analysis revealed that the total treatment time of IVVH( OR=1.067, P<0.001), VTI( OR=0.652, P=0.024), CO( OR=0.037, P<0.001), and RI(OR<0.001, P=0.010)of the interlobar artery were identified as independent factors influencing the recovery of renal function in AKI patients at 30 days and 90 days after IVVH treatment.The ROC curve demonstrated the predictive value of various independent influencing factors for 30-day renal function recovery.The area under the curve(AUC)for VTI was 0.610(95% CI: 0.513-0.707), for CO it was 0.760(95% CI: 0.656-0.864), and for RI it was 0.694(95% CI: 0.589-0.799).Similarly, the ROC curve showed the predictive value of these factors for renal function recovery at 90 days.The AUC for VTI was 0.654(95% CI: 0.564-0.744), for CO it was 0.697(95% CI: 0.605-0.789), and for interlobar artery RI it was 0.605(95% CI: 0.495-0.715). Conclusions:The venous transit time index(VTI), cardiac output(CO), and renal interlobar artery RI, monitored by critical care ultrasound, are independent factors that can be used to evaluate the recovery of renal function in elderly patients with HF and AKI after IVVH treatment.Additionally, the changes in these parameters within 7 days after IVVH treatment have a high predictive value for the improvement of renal function in elderly patients after 30 days and 90 days.

Objective:To investigate the volume management of intermittent veno-venous hemofiltration (IVVH) guided by critical care ultrasound in the treatment of acute kidney injury (AKI) in patients with heart failure (HF).Methods:A total of 216 patients with HF and AKI treated with IVVH in the coronary care unit (CCU) of the Third Central Hospital of Tianjin from April 2019 to June 2022 were selected as the study subjects, the patients were randomly divided into conventional guidance group (107 cases) and ultrasound guidance group (109 cases). According to the recovery of renal function, IVVH was performed 12 hours every day or 12 hours every other day. The conventional guidance group selected the conventional method to formulate IVVH prescription, and the ultrasound guidance group used critical care ultrasound to adjust the treatment parameters of IVVH on the basis of the conventional guidance group. Respiratory variation index (RVI) of inferior vena cava (IVC), right left ventricular end-diastolic transverse area ratio, early diastolic peak mitral flow velocity/mitral annulus velocity peak (E/E'), aortic flow velocity time integral (VTI), cardiac output (CO), bilateral lung ultrasound B-line range, bilateral renal interlobar arteries resistance index (RI) were recorded before and 3, 6, 9 hours after each treatment. The net dehydration rate was adjusted in real time according to the comprehensive results. Urine volume, serum creatinine (SCr), estimated glomerular filtration rate (eGFR), blood B-type brain natriuretic peptide (BNP), β 2-microglobulin (β 2-MG) and cystatin C (Cys C) levels of patients in both groups were monitored before and 3, 7 and 10 days after initial treatment, and renal function recovery and clinical prognostic indexes of patients in both groups were recorded. Results:The dehydration rate of the ultrasound guidance group was slow at the beginning of IVVH, and gradually increased after 6 hours, and the overall dehydration rate was significantly slower than that of the conventional guidance group. In the ultrasound guidance group using critical care ultrasound, the RVI gradually increased, the right left ventricular end-diastolic area ratio gradually decreased, the E/E' ratio gradually decreased, and the range of B-line of bilateral lungs gradually decreased, RI of bilateral renal interlobar arteries decreased. At 3, 7 and 10 days after the first IVVH, renal function related indexes in both groups were significantly improved compared with before treatment, and the decline rate of β 2-MG and Cys C in the ultrasound guidance group was faster than that in the conventional guidance group at early (3 days) [β 2-MG (mg/L): 3.69±1.31 vs. 3.99±1.45, Cys C (mg/L): 2.91±0.95 vs. 3.14±0.96, both P < 0.05], urine volume, SCr and eGFR at 7 days were also significantly improved compared with the conventional guidance group [24-hour urine volume (mL): 1 128.23±153.92 vs. 1 015.01±114.18, SCr (μmol/L): 145.86±32.25 vs. 155.64±28.42, eGFR (mL/min): 50.26±11.24 vs. 46.51±10.61, all P < 0.05]. The time of SCr recovery, the time of reaching polyuria, the total time of IVVH treatment, the time of non-invasive mechanical ventilation and the time of living in CCU in the ultrasound guidance group were shorter than those in the conventional guidance group. The incidences of hypotension, long-term RRT, incidence of major cardiovascular adverse event (MACE) and at 28-day mortality were all lower than those in the conventional guidance group. Kaplan-Meier survival curve showed that the 28-day cumulative survival rate in the ultrasound guidance group was significantly lower than that in the conventional guidance group (Log-Rank test: χ 2 = 3.903, P = 0.048). Conclusion:The strategy of IVVH guided by critical care ultrasound in the treatment of HF with AKI has unique advantages.

AIM: To investigate the influence of mesenchymal stem cells (MSCs) expressing indoleamine 2, 3-dioxygenase (IDO) activity on the allogeneic T-lymphocyte responses.METHODS: MSCs were isolated and cultured from human bone marrow. Selected surface markers of MSCs were detected by flow cytometry and their morphologic characteristics were determined by microscopy. The pluripotentiality of MSCs was also studied. IDO mRNA and IDO protein expressions in MSCs induced by γ-interferon (IFN-γ) at the concentration of 2×105 U/L were detected. MSCs induced by IFN-γ at the concentration of 2×105 U/L were plated in dishes and then mixed lymphocyte reaction (MLR) cultures were set up. T-lymphocytes proliferation was determined by MTT assays and IDO activity was measured by high-performance liquid chromatography (HPLC). RESULTS: IFN-γ could stimulate IDO mRNA and IDO protein expressions in MSCs. IDO enzyme activity in induced MSCs inhibited T-lymphocyte proliferation of MLR cultures. CONCLUSION: MSCs could suppress T-lymphocyte responses via IDO enzyme activity in vitro.

The immune mechanism of Graves' diseases (GD) and the roles of regulator T cells were investigated. In 32 patients with GD (GD group) and 20 healthy volunteers (control group), flow cytometry was used to detect the proportion of CD4+CD25+ cells, MACS to isolate CD4+ CD25+ cells, RT-PCR to assay the expression of FOXP3, and ELISA to test the level of IL-10, respectively. It was found that there was no significant change in the proportion of CD4+CD25+ T cells between GD group and control group (P>0.05), while secretion of IL-10 and expression of FOXP3 in GD group were lower than control group (P<0.01 and P<0.05, respectively). In conclusion, though the proportion of regulatory T cells of peripheral blood lymphocytes in the patients with GD, the functions of them were significantly weakened, which might be a pathogenic factor in GD.

Objective To investigate the influence of indoleamine 2, 3-dioxygenase (IDO) (activity) from mesenchymal stem cells (MSCs) on the allogeneic T-lymphocytes responses.Methods MSCs were isolated and cultured from human bone marrow. Selected surface antigens of MSCs were detected by flow cytometry and their morphologic characteristics were determined by microscopy. In MSCs induced by ?-interferon (IFN-?) at different concentrations (0, 20, 50, 100, 200 U/ml), IDO mRNA expression and IDO activity were detected. MSCs Induced by IFN-? at the concentration of 200 U/ml were plated in dishes and then mixed lymphocyte reaction (MLR) cultures were set up. T- (lymphocytes) proliferation was determined by MTT assays and IDO activity was determined by high-performance liquid chromatography (HPLC).Results IFN-? could stimulate the IDO mRNA expression and IDO activity in MSCs in a dose-dependent manner. IDO enzyme activity in MSCs inhibited T-lymphocytes proliferation of MLR cultures.Conclusion MSCs could suppress T-lymphocyte responses via IDO enzyme activity in vitro.

AIM: To study the biology characteristics of mesenthymal stem cells(MSCs) derived from chronic myelogenous leukemia(CML) and normal adult bone marrow.METHODS: Mononuclear cells from chronic myelogenous leukemia(n=19) and normal adult(n=8) bone marrow were obtained,cultured in expanded medium with low serum concentration.Cell morphology,cell cycle,immunophenotype and in vitro differentiation capacity were investigated.The differentiations of osteocytes and adipocytes were detected by von Kossa staining and Oilred O staining.The chimeric oncogene BCR/ABL and Ph chromosome,two hall marks of CML,were detected in CML derived MSCs,normal adult MSCs,CML derived hematopoietic cells and K562 cells.RESULTS: CML and normal adult derived MSCs showed similar characteristics in cell morphology,phenotype and growth pattern.A typital fibrablast like morphology was observed.Under suitable conditions,CML and normal adult MSCs had the similar ability to differentiate into adipocytes and osteoblasts in vitro.Moreover,CML and normal adult MSCs did not express BCR/ABL gene products and Ph chromosome was not observed.CONCLUSIONS: We isolated and cultured a population of cells with characteristics of multipotent stem cells from CML bone marrow.There were similar biologic characteristics and differentiation ability between normal adult and CML bone marrow-derived MSCs.

AIM: To investigate the isolation,purification,expansion and multilineage differentiation of mesenchymal stem cells(MSCs) derived from human umbilical cord vein in vitro.METHODS: By 1% collagenase Ⅱ digestion,endothelial cells were isolated from human umbilical cord vein and cultured in IMDM medium.The morphology of the cells was observed by Wright's staining and electron microscope.Cell cycle and immunophenotype were investigated by flow cytometry.Assays of adipogenic and osteogenic differentiation were performed in vitro.von Kossa staining,Oil Red O staining and mRNA expression of osteopontin and lipoprotein lipase were studied in the induced cells.RESULTS: The cells from the cord vein displayed a fibroblast-like morphology adhering to the culture plate.FACS showed that the cells expressed several MSCs-related antigens such as CD29,CD44 and CD105,while CD13,CD31,CD45,CD34,and HLA-DR were negative.Adipocyte and osteocyte differentiation were induced successfully.CONCLUSION: The morphology,growth characteristics,immunophenotype and pluripotentiality of the MSCs from human umbilical cord vein are similar to the MSCs from bone marrow(BM).They could potentially be an excellent source of MSCs for experiments and clinics.