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BACKGROUND:The research of dental stem cells in the fields of regenerative medicine and tissue engineering has been deepening,bringing hope for the repair of tooth-related tissues and the treatment of systemic diseases.However,there is a lack of systematic research and analysis on the biological characteristics of dental stem cells in different age groups. OBJECTIVE:To explore the biological characteristics of the human deciduous tooth and permanent tooth pulp stem cells cultured in umbilical cord blood platelet lysate to provide a reliable basis for human platelet lysates to replace fetal bovine serum. METHODS:The pulp tissues of deciduous teeth,juvenile permanent teeth and adult permanent teeth were taken out and cultured in DMEM/F-12 medium supplemented with 10%fetal bovine serum or different concentrations(5%,10%and 15%)of human platelet lysates.Cell proliferation in the four groups was detected by cytometry.The optimal concentration of human platelet lysates was selected for subsequent experiments.Under the optimal concentration of human platelet lysates,human deciduous tooth and juvenile and adult permanent tooth pulp stem cells were cultured in vitro.The cell growth status was observed under the microscope.The specific antigen on the cell surface was detected by flow cytometry.The cell proliferation ability was tested by the cell counting method and CCK-8 assay.The cell differentiation ability in vitro was observed by a three-line differentiation assay. RESULTS AND CONCLUSION:(1)The cell proliferation rate of the 10%human platelet lysate group was the highest.(2)In all three groups,fusiform fibrous cells grew and expanded from around the tissue block.There was no significant difference between deciduous teeth and juvenile permanent tooth cells,but the adult permanent tooth cells were larger than the deciduous and juvenile permanent tooth cells of the same generation.(3)The results of flow cytometry showed that deciduous teeth,juvenile permanent teeth and adult permanent teeth conformed to the phenotypic characteristics of mesenchymal stem cells.(4)The proliferative capacity of adult permanent dental pulp stem cells was significantly lower than those of deciduous teeth and juvenile permanent dental pulp stem cells(P<0.01).(5)mRNA expressions of osteoblast-related genes alkaline phosphatase and bone morphogenetic protein 2,lipoprotein lipase and peroxisome proliferator-activated receptor γ2,mRNA expressions of chondroblast related gene type II collagen α1 and cartilage oligomeric matrix protein in adult pulp stem cells of permanent teeth were significantly lower than those of deciduous teeth and juvenile permanent teeth pulp stem cells(P<0.01).(6)Compared with adult dental pulp stem cells,human deciduous teeth and juvenile permanent teeth dental pulp stem cells have the stronger proliferative capacity and multidirectional differentiation potential,and are more suitable for clinical research and disease treatment.
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【Objective】 To investigate the detection of pathogenic microorganisms in umbilical cord blood and maternal blood from 2012 to 2021, so as to improve the collection of umbilical cord blood and guarantee the safety of umbilical cord blood hematopoietic stem cells (HSC) . 【Methods】 Detection results of pathogenic microorganisms of umbilical cord blood and maternal blood among 64 077 cases from Tianjin Cord Blood Bank from 2012 to 2021 were retrospectively analyzed. 【Results】 A total of, 2 072 cases (3.23%) were detected positive, among which, 184 cases (0.29%) were positive for aerobic bacteria culture, 1 504 cases (2.34%) were positive for anaerobic bacteria culture, and 384 cases (0.60%) were positive for both aerobic and anaerobic bacteria culture. From 2012 to 2021,the overall positive rate showed a downward trend, with a difference in the positive rate between each year (P<0.05). The positive rate of anaerobic bacteria was higher than that of aerobic bacteria and that of anaerobic and aerobic bacteria (P<0.05). After Gram staining, the microscopic detection rate of bacterial positive samples was highest in G- bacilli, followed by G+ bacilli, G+ cocci, G- cocci and others. Among the 64 077 cases, 169 cases (0.26%) showed reactivity in cord blood tests and 1 231 cases (1.92%) showed reactivity in maternal blood tests. Umbilical cord blood and maternal blood HIV-Ag/Ab tests showed reactivity after initial screening. After confirmation by Western blotting, there was 1 case of uncertain maternal blood, while the rest were negative. The reactive rates of anti-TP (0.12%) and anti- HCV (0.11%) in umbilical cord blood were higher than those of HBsAg (0.03%) and CMV-IgM (1/64 077).There was a difference in the reactive rate of anti-TP detection in umbilical cord blood between different years (P<0.05),while there was no statistically significant difference in that of HBsAg, anti-HCV and CMV-IgM (P> 0.05).The reactive rate of HBsAg in maternal blood (1.38%) was higher than that of CMV-IgM(0.29%) , anti-TP(0.13%) and anti-HCV (0.12%) . There were differences in the reactive rates of HBsAg, anti-HCV ,and anti-TP in maternal blood among different years (P<0.05),and that of HBsAg showed a decreasing trend, while the reactive rate of CMV-IgM was not statistically significant (P>0.05). The reactive rates of HBsAg and CMV-IgM detected in maternal blood were significantly higher than those in umbilical cord blood (P<0.05) . The reactive rates of anti-HCV and anti-TP in maternal blood were consistent with those in umbilical cord blood (P>0.05). 【Conclusion】 The reactive rates of anti-HIV and CMV-IgM in cord blood, and that of anti-HIV in maternal blood are low, but those of anti-TP and anti-HCV in cord blood are relatively high. The reactive rate of HBsAg is high in maternal blood,but with a downward trend,but low in umbilical cord blood due to maternal-infantile transmission blocking. The detection of transfusion transmitted pathogens and bacteria plays a critical role on the safety of umbilical cord blood HSCs. Effective detection of transfusion transmitted pathogens and culture of bacteria are the key to ensure the quality of umbilical cord blood, which can improve the safety of umbilical cord blood HSCs transplantation.
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Objective:To investigate the level of trimethylamine N-oxide (TMAO), one of gut metabolites, in patients undergoing maintenance hemodialysis (MHD) accompanied by congestive heart failure (HF) and its influencing factors.Methods:Those patients of 18-75 years old who received three or more times of hemodialysis sessions per week for three months or longer during Nov 2018 and Mar 2019 were enrolled. Those attended health checkup at the same time without obvious kidney abnormality served as non-kidney disease controls. Serum TMAO concentrations were measured using high-performance liquid chromatography electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS). The levels of TMAO were compared between patients on hemodialysis and controls, between those with heart failure and without heart failure using logrithmically transformed TMAO (lnTMAO). Linear regression analysis was performed to investigate factors influencing TMAO levels.Results:A total of 195 patients undergoing MHD and 40 controls were enrolled. Among them, 30 hemodialysis cases (15.4%) manifested as heart failure symptoms and/or left ventricular ejection fraction less than 50%. Males accounted for 67.2% in patients on hemodialysis and 37.5% in controls ( χ2=12.426, P<0.001) respectively, while the median ages in both groups were 62.0(48.0, 71.0), 45.0(33.3, 55.0) years old respectively ( Z=5.685, P<0.001). TMAO concentrations were significantly higher in patients on hemodialysis than controls [5.54(3.84, 8.91) mg/L vs 0.17(0.11, 0.30) mg/L, after log transformed, t=21.687, P<0.001]. However, there was no statistically significant difference between those with heat failure and those without in male [63.3% vs 67.9%, χ2=0.238, P=0.626], age [64.5(56.8, 71.0) years old vs 61.0(47.0, 72.0) years old, Z=0.894, P=0.372] and TMAO [5.17(3.30, 9.46) mg/L vs 5.57(3.87, 8.95) mg/L, after log transformed, t=-1.537, P=0.135]. Multivariate linear regression analysis demonstrated that in all the participants, serum urea was the main risk factor for TMAO [standardized coefficient ( SB)=0.483]. lnTMAO=0.078×[serum urea(mmol/L)]+0.001×[serum creatinine (μmol/L)]-0.002×[serum uric acid (μmol/L)]-0.003×[platelet (×10 9/L)]+0.014×[age (years old)]+0.344 (if diabetic)-1.266. While in those undergoing MHD, ultrafiltration volume had the most significant effect on TMAO levels ( SB=0.279). lnTMAO=0.249×[ultrafiltration volume(L)]+0.059×[serum albumin (g/L)]+0.008×[age (years old)-0.526 (if heart failure existed)-1.865. Conclusions:MHD patients have gut dysbiosis, while those hemodialysis patients accompanied by heart failure may have peculiar gut microbiota which induces lower serum TMAO levels than those without heart failure after adjusting for multiple related factors. Serum TMAO levels may be associated with ultrafiltration volume and nutrition status etc.
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Related substances in pharmaceutical formulations are associated with their safety, efficacy and stability. However, there is no overall study already published on the assessment of related substances in the Compound Ketoconazole and Clobetasol Propionate Cream. In this work, a reliable HPLC-TOF-MS qua-litative method was developed for the analysis of related substances in this preparation with a quick and easy extraction procedure. Besides the active pharmaceutical ingredients, two compounds named ke-toconazole impurity B′ optical isomer and ketoconazole impurity E were identified. Furthermore, a new HPLC method for qualitative and quantitative assessment on related substances and degradation pro-ducts, which were found in the stability test, was established and validated. The single standard to determine multi-components method was applied in the quantitative analysis, which was an effective way for reducing cost and improving accuracy. This study can provide a creative idea for routine analysis of quality control of the Compound Ketoconazole and Clobetasol Propionate Cream.
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BACKGROUND: A large amount of studies have demonstrated that stem cells in the heart of mammal participate in heart self-renewal and endogenous repair.OBJECTIVE: To summarize the source, classification, features of cardiac stem cells and application in heart disease. METHODS: A computer-based online search of PubMed database was performed for articles published between January 2000 and December 2010 related to the source, classification, features of cardiac stem cells and its effects on myocardial regeneration with key words cardiac stem cell. Clinical studies and basic studies were all included. Repetitive studies and Meta analysis were excluded. Finally, 32 articles were included.RESULTS AND CONCLUSION: Cardiac stem cell is a type of stem cells in the heart, with properties of self-renewal and cloning proliferation. It can differentiate into cardiomyocyte and endothelial cell and plays a role in heart injury repair to improve heart function. Cardiac stem cells have been in vitro isolated, cultured and amplified to transplant into animal heart, which lays a foundation for clinical application. However, some questions remain poorly understood as to the homeostasis and dynamic changes, processes involved in their differentiation into heart functioning cells, as well as influential factors and regulation.