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Aging-elevated DNMT3A R882H-driven clonal hematopoiesis (CH) is a risk factor for myeloid malignancies remission and overall survival. Although some studies were conducted to investigate this phenomenon, the exact mechanism is still under debate. In this study, we observed that DNMT3A R878H bone marrow cells (human allele: DNMT3A R882H) displayed enhanced reconstitution capacity in aged bone marrow milieu and upon inflammatory insult. DNMT3A R878H protects hematopoietic stem and progenitor cells from the damage induced by chronic inflammation, especially TNFα insults. Mechanistically, we identified that RIPK1-RIPK3-MLKL-mediated necroptosis signaling was compromised in R878H cells in response to proliferation stress and TNFα insults. Briefly, we elucidated the molecular mechanism driving DNMT3A R878H-based clonal hematopoiesis, which raises clinical value for treating DNMT3A R882H-driven clonal hematopoiesis and myeloid malignancies with aging.
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Objective To investigate the radioprotective function and its mechanism of miR-223 in acute radiation-induced lung injury in mice.Methods Forty female C57BL/6 J mice were randomly divided into healthy control group,irradiation group,irradiation plus miR-223 group and irradiation plus NC group.Radiation groups were exposed with a single dose of 15 Gy of 6 MV X-rays delivered by a linear accelerator.The mice in drug group were administered by tail vein injection with miR-223 agomir or agomirNC every other day from 1 d before irradiation to 14 d after irradiation.The lung tissue samples of mice were taken at 14 d post-irradiation.The pathological changes were observed by HE staining.The localization and expressions of IL-1β and IL-18 were observed by immunohistochemistry (IHC).Real-time PCR was used to detect miR-223,but NLRP3 mRNA expression in lung tissue.Western blot was used to detect the protein expressions of NLRP3 and Caspase-1,and ELISA assay was used to detect the expressions of IL-1β and IL-18 in lung homogenate.Results Radiation decreased the expression of miR-223,but increased the expression of NLRP3 in lung tissue.Administration of miR-223 agomir inhibited the expression of NLRP3 and attenuated lung inflammation.HE and IHC staining showed that miR-223 reduced the acute inflammatory response and the expressions of IL-1β and IL-18 in lung tissue compared with irradiation group (t=10.16,6.00,P＜0.05).The expressions of NLRP3 and Caspase-1 protein in lung tissue of irradiated plus miR-223 group was lower than that in the irradiation alone group (t =12.47,4.95,P＜ 0.05).ELISA assay also showed a decrease of inflammatory factors IL-1β and IL-18 in lung tissue homogenate of the irradiation plus miR-223 group (t =8.22,8.47,P＜0.05).Conclusions MiR-223 effectively reduces the secretion of radiation-induced inflammatory factors IL-1β and IL-18 by inhibiting the expression of NLRP3 in lung tissue of mice,and thus has protective effect on radiation-induced lung injury.
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Objective@#To investigate the effect of ubiquitous mitochondrial creatine kinase 1(CKMT1) on the sensitivity of human nasopharyngeal carcinoma cell line CNE-1 to DDP. @*Methods@#CNE-1 cells were transiently transfected with CKMT1 overexpression (CKMT1) or empty vector (EV). The growth curve and DDP IC50 were developed by MTT assay, plate clone formation assay was performed by gradient concentration of DDP treatment, cell cycle and apoptosis were detected by flow cytometry, levels of apoptosis related protein Bax/Bcl-2/C-PARP and the transcription factor p-STAT3-Tyr705 were detected by Western Blot. @*Results@#The transfection efficiencies of CKMT1 and EV were more than 90% with a higher proliferation rate in the CKMT1-transfected cells. However, the CKMT1-transfected cells had a DDP IC50 of 2.76 μmol/L, which was significantly lower than that of 4.60 μmol/L in the EV-transfected cells (P<0.01). With the treatment of certain concentration of DDP, the CKMT1-transfected cells had a lower clone formation rate, the cell cycle arrested more obviously in G2/M phase, and the apoptosis rate was higher (P<0.01), with higher levels of Bax/C-PARP (P<0.05 or P<0.01), but lower levels of Bcl-2 (P<0.01) and p-STAT3-Tyr705 (P<0.01), compare with the EV-transfected cells. @*Conclusions@#CKMT1 may inhibit the activation of STAT3, increasing the sensitivity of CNE-1 to chemotherapeutic drug DDP.
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OBJECTIVE@#To investigate androgen receptor (AR) expression and the effect of epidermal growth factor (EGF) and testosterone on AR expression level. @*METHODS@#EGF or different concentrations of testosterone were incubated with the primary urethral plate fibroblasts from patients with hypospadias. The levels of AR expression in the fibroblasts were detected by immunocytochemical assays and graphical analysis. @*RESULTS@#There was no significant difference in AR activation under physiological concentrations (3×10(-8) mol/L) of testosterone between the control and the distal hypospadias group (P>0.05). However, there was a significant decrease in AR activation in the proximal hypospadias group compared to that in the control group (Pdistal hypospadias group>proximal hypospadias group, P<0.001). AR activation level in the group of proximal hypospadias was improved most obviously when EGF and physiological concentration of testosterone were employed in the urethral plate fibroblasts from hypospadias patients (P<0.001), and it was improved more in the distal hypospadias group than that in the control group (P=0.02). @*CONCLUSION@#AR expression and activation in the urethral plate fibroblasts from hypospadias patients are abnormal. EGF can be used to improve AR activation in fibroblasts from different types of hypospadias, especially in the proximal type.
Subject(s)
Humans , Male , Cells, Cultured , EGF Family of Proteins , Metabolism , Fibroblasts , Metabolism , Hypospadias , Metabolism , Receptors, Androgen , Metabolism , Testosterone , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the dynamic changes in CD4(+)CD25(+) regulatory T cells and Foxp3 expression in peripheral blood and brain tissues of rats after acute cerebral ischemia and explore their role in the pathophysiological evolution of acute ischemic stroke.</p><p><b>METHODS</b>Forty-eight Wistar rats were randomized equally into ischemia and sham-operated groups, and right middle cerebral artery occlusion was induced in the former group. Flow cytometry and immunohistochemistry were employed to detect CD4(+)CD25(+) T cells and Foxp3 expression, respectively, in the peripheral blood and brain tissue at 1, 3, 7, and 14 days after modeling. The behavioral changes of the rats were evaluated using an improved NSS neurological functional scoring system.</p><p><b>RESULTS</b>The neurological function scores of the two groups both gradually declined after the operation, and showed significant differences between the two groups at all the time points of measurement (P<0.01). The CD4(+)CD25 T cells in the peripheral blood were similar between the two group at 1 and 3 days after the operation (P>0.05), but increased significantly in the ischemia group at 7 and 14 days (P<0.05) with an inverse correlation to the neurological scores (r=-0.68, P=0.01). Immunohistochemistry detected the presence of Foxp3 primarily in the ischemic region of the brain tissue 1 day after cerebral ischemia; the contralateral hemisphere also showed a small quantity of Foxp3 expression. No Foxp3 expression was detected in the brain tissue of the sham-operated group.</p><p><b>CONCLUSION</b>CD4(+)CD25 T regulatory cells participate in the inflammatory immune reactions as early as 1 day after acute cerebral ischemia in rats, which might be a protective mechanism of the brain cells.</p>
Subject(s)
Animals , Male , Rats , Brain , Metabolism , Brain Ischemia , Allergy and Immunology , Metabolism , Forkhead Transcription Factors , Allergy and Immunology , Metabolism , Rats, Wistar , T-Lymphocytes, Regulatory , Allergy and Immunology , MetabolismABSTRACT
This paper reviews the effects of physical environments (including light, electric field, ultrasound, magnetic field, microgravity, temperature, mechanical vibration, and heterogeneous nucleation interface) on protein crystal nucleation. The research results are summarized and the possible mechanisms of the effects are discussed. In the end of this review, the application prospects of these physical environments (including coupled environments) in protein crystallization are presented.