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Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs), a kind of adult stem cell, were studied for clinical applications in regenerative medicine. To date, the safety evaluations of intravenous infusion of allogeneic hUC-MSCs were focused on fever, infection, malignancy, and death. However, the characteristics of dynamical changes in vital signs during hUC-MSCs infusion are largely unknown. In this study, twenty participants with allogeneic hUC-MSCs transplanted (MSC group) and twenty sex- and age-matched individuals with cardiovascular disease who treated with the equal volume of 0.9% normal saline were recruited (NS group). Heart rate, respiratory rate, oxygen saturation, systolic and diastolic blood pressure, and temperature were monitored at intervals of 15 min during infusion. Adverse events were recorded during infusion and within seven days after infusion. No adverse events were observed during and after infusion in both groups. Compared with the baseline, the mean systolic blood pressure (SBP) levels were significantly decreased at 15 min, 30 min, 45 min and 60 min in the MSC group (all P < 0.05) during infusion. In addition, SBP changed significantly from baseline during hUC-MSCs infusion when compared with that of NS group (P < 0.05). Repeated measures analysis of variance confirmed difference over time on the SBP levels (P < 0.05). Our results showed that the process of allogeneic hUC-MSCs intravenous infusion was safe and the vital signs were stable, whereas a slight decrease in SBP was observed.
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BACKGROUND: Adipose tissue-derived stromal cells (ADSCs) have been extensively used in clinical trials for various therapeutic applications. However, there is a paucity of selective criteria regarding collection and expansion procedures. The purpose of this study was to investigate the effect of liposuction and donor age on ADSCs and to assess the criteria and markers for ADSC long-term expansion potential. METHODS: Adipose tissues were collected using syringe liposuction, water-jet, or ultrasonic techniques. Tissue/cell viability was evaluated using the XTT assay. CD34 and SSEA-4 expression were examined using flow cytometry. SOX2 gene expression was estimated using quantitative polymerase chain reaction, and Nile-red staining was performed to evaluate the adipogenesis potency during ADSC expansion. RESULTS: The lipoaspirates obtained from syringe and ultrasonic liposuction methods were superior to those of the water-jet method in stromal vascular fraction yield and durability during temporary storage. SSEA-4, SOX2 expression, and adipogenesis potency of early-passage ADSCs were significantly correlated with the P15 cumulative population doublings data. CD34 expression was strongly correlated with P0 ADSC yield and doubling time. Tissue viability, P0 ADSC CD34⺠percentage, and P15 cumulative population doublings were decreased along with donor age. CONCLUSIONS: This study established criteria and markers to determine whether lipoaspirate tissue and cultured ADSCs are suitable for further large-scale expansion and allogenic universal cell banking.
Assuntos
Seleção do Doador/métodos , Lipectomia/métodos , Células-Tronco Mesenquimais/fisiologia , Gordura Subcutânea/citologia , Preservação de Tecido/métodos , Adulto , Fatores Etários , Idoso , Biomarcadores/metabolismo , Sobrevivência Celular , Células Cultivadas , Feminino , Citometria de Fluxo , Humanos , Masculino , Células-Tronco Mesenquimais/metabolismo , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
BACKGROUND: Cardiac cellular injury as a consequence of ischemia and reperfusion involves nuclear factor-κB (NF-κ B), amongst other factors, and NF-κ B inhibitors could substantially reduce myocardial infarct size. Parthenolide, a sesquiterpene lactone compound which could inhibit NF-κ B, has been shown to ameliorate myocardial reperfusion injury but may also produce toxic effects in cardiomyocytes at high concentrations. The aim of this study was to examine the cytotoxic effects of this drug on H9c2 cardiomyoblasts, which are precursor cells of cardiomyocytes. METHODS: Cell viability and apoptosis were examined by MTT and TUNEL assay, respectively, and protein expression was analyzed by western blot. Reactive oxygen species (ROS) production was measured using DCFH-DA as dye. Cytosolic Ca(2+) concentration and mitochondrial membrane potential were measured microfluorimetrically using, respectively, fura 2 and rhodamine 123 as dyes. RESULTS: Parthenolide caused apoptosis at 30 µ M, as judged by TUNEL assay and Bax and cytochrome c translocation. It also caused collapse of mitochondrial membrane potential and endoplasmic reticulum stress. Parthenolide triggered ROS formation, and vitamin C (antioxidant) partially alleviated parthenolide-induced cell death. CONCLUSIONS: The results suggested that parthenolide at high concentrations caused cytotoxicity in cardiomyoblasts in part by inducing oxidative stress, and demonstrated the imperative for cautious and appropriate use of this agent in cardioprotection. KEY WORDS: Cardiomyoblast; Endoplasmic reticulum stress; Oxidative stress; Parthenolide; Reperfusion injury.
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BACKGROUND: Strong P2X7 receptor (P2X7R) activation causes Ca(2+) overload and consequent cell death. We previously showed that depletion of Ca(2+) stores and endoplasmic reticulum (ER) stress in differentiated NG108-15 neuronal cells contributed to P2X7R-mediated cytotoxicity. In this work, we assessed whether taurine (2-aminoethanesulfonic acid) could prevent this P2X7R-mediated cytotoxicity in this neuronal cell line. METHODS: Cytotoxicity markers were assessed by MTT assay and Western blotting. Cytosolic Ca(2+) and mitochondrial Ca(2+) concentrations were measured microfluorimetrically using fura-2 and rhod-2, respectively. Intracellular reactive oxygen species (ROS) production was assayed by the indicator 2',7'-dichlorodihydrofluorescein diacetate. RESULTS: Selective P2X7R agonist BzATP treatment causes neuronal cell death by causing cytosolic Ca(2+) overload, depletion of Ca(2+) stores, endoplasmic reticulum (ER) stress, and caspase-3 activation (cleaved caspase 3). Remarkably, taurine (10mM) pretreatment could prevent P2X7R-mediated neuronal cell death by blocking BzATP-mediated ER stress as determined by phosphorylated eukaryotic translation initiation factor 2α (peIF2α) and C/EBP-homologous protein (CHOP). However, taurine did not block BzATP-induced Ca(2+) overload and depletion of ER Ca(2+) stores. Interestingly, P2X7R activation did not result in mitochondrial Ca(2+) overload, nor did it affect mitochondrial membrane potential. BzATP-induced generation of intracellular reactive oxygen species (ROS) was prevented by taurine. CONCLUSIONS: The neuroprotective effect by taurine is attributed to the suppression of P2X7R-mediated ER stress and ROS formation.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Receptores Purinérgicos P2X7/metabolismo , Taurina/farmacologia , Trifosfato de Adenosina/análogos & derivados , Trifosfato de Adenosina/toxicidade , Animais , Cálcio/metabolismo , Técnicas de Cultura de Células , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Neurônios/citologia , Neurônios/metabolismo , Agonistas do Receptor Purinérgico P2X/toxicidade , Espécies Reativas de Oxigênio/metabolismoRESUMO
AIM: To observe the effects of Compound Muji Powder (CMJP) on chronic patients with hepatitis B. METHODS: 28 chronic patients with hepatitis B were chosen as therapy group who took CMJP and general protecting liver medicine, another 31 patients taking general protecting liver medicine only were taken as control group. Clinic symptom, liver function, serum fibrosis items, parts of immunological items, blood routine test and ultrasound alteration were observed before, after and 3 months after ending of treatment and compared between the two groups. RESULTS: Many detecting items including ALT, AST, r-GT, HA, LN, PCIII, IgG, r-globin, AFP, PLT, thickness of spleen and echogenicity of liver had changed obviously after CMJP application, and parts of these alteration maintained rather long time. CONCLUSION: Since CMJP could play the role of protecting the liver cells, inhibiting liver fibrosis, decreasing AFP level and reducing the enlargement of spleen, it had good curative effects to chronic hepatitis B patients.