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OBJECTIVE: To investigate the effects of resveratrol (Res) on aging of marrow mesenchymal stem cells (MSCs), and to explore its mechanism. METHODS: MSCs were isolated from young SD rats and cultured in vitro. The optimal D-gal concentration for induction of MSCs senescence was determined. Then MSCs were randomly divided into four groups, namely the control group, 10μmol/L, 50μmol/L and 100μmol/L Res groups. After the cells were treated with different concentration of Res for 48 h, the senescence-associated changes were examined with senescence-associated-β-galactosidase (SA-β-gal) staining; the expression of p53, p16 and γ-H2AX was evaluated by Western blot. The total active oxygen species (ROS) level was determined by flow cytometry with DCFH-DA staining. In order to assess the effect of Res on the mitochondrial function, MitoSox Red staining was used to detect mitochondrial ROS levels in each group, mitochondrial membrane potential was detected by JC-1 assay, mPTP method was used to detect mitochondrial membrane channel opening level, and Western blot was used to detect the expression level of cytoplasmic cytochrome C (Cyt-C). RESULTS D-gal 10 and 50 g/L significantly increased the number of SA-β-gal positive cells and the level of mitochondrial ROS (all P<0.01). Therefore, 10 g/L D-gal was used to induce the senescence of MSCs in subsequent experiment. Compared with the control group, the number of SA-β-gal positive cells in Res groups significantly decreased (all P<0.01), the expression of p53, p16 and γ-H2AX decreased, and the total and mitochondrial ROS level also decreased (all P<0.01). Moreover, mitochondrial membrane potential, open level of mitochondrial membrane channels and the levels of cytoplasm Cyt-C in the Res treatment groups decreased compared with the control group (P<0.05 or P<0.01). CONCLUSIONS Resveratrol can protect the mitochondrial function of MSCs, and effectively delay the MSC senescence.
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Objective@#To understand the current situation of sedentary behaviors among middle school students and its relationship with physical health.@*Methods@#Data came from the Taizhou City and Township Health Survey in 2018. Sedentary behaviors and physical fitness of 2 374 middle school students in Taizhou were analyzed.@*Results@#The total time spent in sedentary behaviors of middle school students in Taizhou was (8.75±1.56) h/d on weekday and (7.34±1.55) h/d on weekend; Compared with students whose screen time <2 h/d in weekday or weekend, weekend homework time <2 h/d and private tutoring class <2 h/d, students whose screen time ≥2 h/d in weekday or weekend, weekend homework ≥2 h/d and private tutoring class ≥2 h/d showed higher rate of low physical fitness(OR was 1.43, 1.37, 1.12, 1.43, respectively, P<0.05).@*Conclusion@#Long duration of weekend homework and private tutoring class, as well as high screen time in weekday and weekend is associated with decline of physical fitness among middle school students.
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This paper describes a practical process for a SGLT2 inhibitor dapagliflozin. The target product was synthesized from 1-chloro-2-( 4-ethoxybenzyl)-4-iodobenzene and 2, 3, 4, 6-tetra-O-acetyl-alpha-D-glucopyranosyl bromide by iodine-magnesium exchange, and coupling and acetyl removing reactions with the total yield of 50%. This practical process highlights fewer reaction steps, less waste and mild reaction conditions.
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Objective To analyze the change of chromosome test level after participating in the inter‐laboratory proficiency test(PT) program of the College of American Pathologists (CAP) .Methods The results for participating in CAP PT during 2011-2014 were analyzed ,the accuracy of the cytogenetic chromosome quality control test in this laboratory was obtained accord‐ing to the CAP evaluation results ,and the effect of CAP external quality assessment item on increasing the chromosome detection level in this laboratory was evaluated by analyzing the chromosome band levels before and after participating in CAP .Results This laboratory participated in CAP PT test for 10 times during 2011-2014 ,a total of 59 cases were analyzed ,the accuracy rate for jud‐ging chromosome karyotype was 100% ,the karyotype description accuracy rate was 95 .1% .The chromosome test results of clinical cases in this laboratory displayed that peripheral blood chromosome abnormal detection rate was 18 .9% and bone marrow chromo‐some abnormal detection rate was 25 .9% ,the abnormal rate of newly diagnosed leukemia was 66 .8% ;the detection failure rates of peripheral blood chromosome and bone marrow chromosome were 0 .5% and 5 .0% respectively ;the detection failure rates of pe‐ripheral blood chromosome and bone marrow chromosome after participating in CAP were decreased ,the chromosome band average level was improved ,showing statistical difference compared with those before participating in CAP (P<0 .01) .Conclusion Partici‐pating in high quality external laboratory assessment item can increase the clinical analytic level of cytogenetic chromosome test .
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Objective To observe the expression changes of peripheral endothelial progenitor cells (EPCs) and Ang-1/Tie2 in patients with pulmonary arterial hypertension (PAH).Methods From Jun 2011 to Dec 2012,45 patients with PAH charged in Affiliated Hospital of Hangzhou Normal University were divided into 3 groups according to mean pulmonary arterial blood pressure (15 per group):mild(Group L),moderate(Group M),and severe(Group S),with another 15 normal people as control group(Group C).The EPCs were isolated from peripheral blood of every patient,number counting using fluorescence activated cell sorter (FACS),function test using cell culture in vitro.Expression of Ang-1 and Tie2 in the peripheral EPCs were measured by RT-PCR or Western-blot.Non normal data was analyzed by non parametric statistical test.Results The statistical discrepancy existed among Group L,M,S and the control in the number of EPCs [32.0 (27.0,37.0),26.0 (19.0,31.0),24.0 (22.0,26.0) vs 40.0 (37.0,51.0),P ＜ 0.05].The ability of migration [32.1 (26.5,37.5),26.8 (22.4,35.4),21.0 (17.8,34.0) vs 39.0 (33.3,42.4),P＜0.05] and adhesion of the EPCs [57.1(50.9,61.8),51.8(45.2,58.7),46.0 (37.2,55.1) vs 64.1 (56.2,75.0),P ＜ 0.05] among study groups and control group was different in statistic,the same with the proliferation activity of EPCs in different groups [0.6 (0.5,0.7),0.5 (0.4,0.6),0.4(0.3,0.5) vs 0.7(0.6,0.8),P ＜0.05].The mRNA expression of Ang-1 and Tie2 in Group M & S were significantly reduced compared with control [4.33 (2.49,4.62) and 2.89 (2.39,3.44) vs 5.31(3.78,6.22),P＜0.05],Tie2 mRNA[1.32(1.23,1.34)and 1.23(1.08,1.42)vs 1.49(1.25,1.66),P ＜ 0.05],and the protein expression of the phosphorylated Tie 2 in Group M &S were decreased [0.16 (0.15,0.24) and 0.12 (0.08,0.18) vs 0.22 (0.19,0.28),P ＜ 0.05].No significant difference of Ang-1 and Tie2 expression was observed between Group L and control [5.42 (4.72,5.95),1.54 (1.43,1.66) and0.23(0.19,0.33),P=0.674,0.867 and 0.674].Conelusion With the severity of PAH,the number and function of circulating EPCs decreased,as consistent with Ang-1 and Tie2 expression changes,suggesting that function decrease of EPCs in patients with PAH may be associated with the decrease of Ang-1/Tie2 expression.
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Purpose To eva1uate the effects of Annexin A3 on pro1iferation and apoptosis of gastric cancer ce11s. Methods The re-combinant p1asmid pYr-ads-4-Annexin A3 was constructed and ana1yzed by restriction ana1ysis and sequencing and was transfected into MGC803 ce11s. The stab1e transfectants were obtained after screening with G418. Western b1ot ana1ysis was used to examine the expres-sion of Annexin A3 before and after transfection. CCK8 assay,c1one assay and f1ow cytometry were used to study the effects of Annexin A3 on pro1iferation and apoptosis of MGC803 ce11s. Results The recombinant p1asmid pYr-ads-4-Annexin A3 was successfu11y con-structed. Western b1otting resu1ts indicated that the Annexin A3 expression was significant1y higher in ce11s transfected with pYr-ads-4-Annexin A3 compared with ce11s transfected with empty vectors and un-transfected ce11s( P<0. 05 ). CCK8 assay resu1ts showed the number of ce11s transfected with pYr-ads-4-Annexin A3 was significant1y higher than those transfected with empty vectors and un-trans-fected ce11s(P<0. 05). Moreover,the number of c1one in ce11s transfected with pYr-ads-4-Annexin A3 was significant1y higher than the other two groups(P<0. 05). Important1y,high Annexin A3 expression inhibited to apoptosis of MGC803 ce11s(P<0. 05). Con-clusion Annexin A3 expression p1ay important ro1es in tumorigenesis of gastric cancer. Annexin A3 cou1d promote the pro1iferation and inhibited apoptosis of gastric cancer ce11s and it might be a potentia1 target for gastric cancer treatment.
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Rhizoma Pinelliae Fermentata(RPF)wasoneofthecommonlyusedChinesemateriamedica(CMM). According to the ancient and modern literatures on RPF, the historical evolution, fermentation methods, chemical compositions, efficacy and microbes of RPF were systematically summarized in this paper. Through the analysis on existing problems of fermentation strains, effective components, quality standard and fermentation process, the corresponding solutions were proposed. This work may provide an idea and reference for the further study of RPF.
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<p><b>OBJECTIVE</b>To investigate the effects of D-galactose (D-gal) on aging of rat marrow mesenchymal stem cells (MSCs) and its mechanism.</p><p><b>METHODS</b>MSCs isolated from young (7 d) SD rats were randomly divided into four groups:control group, 1g/L, 10g/L and 50g/L D-gal treatment groups. In control group MSCs were cultured in DMEM containing 10% FBS for 48 h. In the D-gal treatment groups, MSCs were cultured in DMEM containing 10% FBS with 1g/L, 10g/L or 50g/L D-gal for 48 h. The senescence-associated changes were examined with SA-β-galactosidase (SA-β-gal) staining, the expressions of p53, p21 and p16 were detected by Western blot. The living and apoptotic cells were determined by AO/EB staining. Cell proliferation was detected by MTT assay. SOD activity was measured by xanthine oxidase method, and the MDA content was estimated with thiobarbituric acid (TBA) method.</p><p><b>RESULTS</b>Compared to control group, the number of SA-β-gal positive cells and the expression of p53, p21 and p16 were significantly increased in the 10g/L and 50g/L D-gal treatment groups. The apoptosis rate in 50g/L D-gal group was significantly higher than that in control group (P<0.01). The proliferation of MSCs was decreased in the 10g/L and 50g/L D-gal groups compared to control group (P<0.05). After 10g/L and 50g/L D-gal treatment, SOD activity was significantly decreased (P<0.01), and MDA level was increased (P<0.01).</p><p><b>CONCLUSION</b>The aging of MSCs can be induced by 10g/L and 50g/L D-gal, which may be associated with the elevated levels of oxidative stress.</p>
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Animals , Male , Rats , Apoptosis , Cells, Cultured , Cellular Senescence , Galactose , Pharmacology , Mesenchymal Stem Cells , Physiology , Oxidative Stress , Rats, Sprague-DawleyABSTRACT
The NF-kappaB pathway regulates the expression of over 150 target genes, e.g., cytokines, chemokines, leukocyte adhesion molecules and inducible effector enzymes. Consequently, it plays a crucial role in innate and adaptive immune responses, inflammatory response, stress responses, apoptosis and so on. IkappaB kinase (IKK) is the key of this pathway, and it owns a special structure which consists of catalytic subunit and regulatory subunit. Naturally, the activation of IKK needs the interaction of the two subunits and phosphorylation by its upstream kinases. Actually, there are two methods of activation of the NF-kappaB pathway, and both of the methods need the IKK complex. Given to the crucial role of IKK, researchers have isolated and synthesized amounts of IKK inhibitors, and these provide a great convenience to develop novel anti-inflammatory and anti-tumor drugs.
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Aim: To synthesize the ent-kaurene diterpenoid and its derivatives from natural available stevioside.Methods: The crucial pharmacophore of exo-methylene ketone was designed and the carboxyl group at C-4 was modified.The cytotoxic inhibition activities in vitro of target compounds were evaluated against human cancer cells BEL-7402,HO-8910,MCF-7 and HL-60 by a MTT method.Results and Conclusion: The struc-tures of new target compounds were identified by ~1H NMR,FT-IR and EI-MS.The preliminary experimental re-sults showed that some derivatives of ent-kaurene diterpenoid possessed fair inhibitory activity against tumor cell lines compared with that of their parent steviol.
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Objective To investigate the changes in morphology and senescence-associated markers of the marrow-derived cardiac stem cells (MCSCs) from rats at different ages and to explore the impacts of age on proliferation, survival and differentiation of MCSCs. Methods With single-cell cloning culture, MCSCs were selected from the bone marrow of young, adult and aged male SD rats respectively. Ultrastructural changes of the cells were viewed under a transmission electron microscope.The senescence-associated changes were examined with SA-β-galactosidase staining and reactive oxygen species(ROS) staining. Distribution of cell cycle of MCSCs from different age groups was evaluated with flow cytometric analysis. Rates of the survived and apoptotic cells were determined by Annexin V/PI double-labeled flow cytometric analysis and Hochest33342 staining. Differentiation of the MCSCs toward cardiomyocytes was induced with BMP-2. Expression of cardiac transcription factors and cardiac specific genes of the cells after induction were examined with RT-PCR. CTnT expression of the cells also be examined with immunocytochemistry. Results The nucleus/plasma ratio of the cells from aged rats decrease and there are some myelin bodies in the cells of aged group. With increasing of age, the MCSCs in S+G2/M phase reduce, while β-galactosidase-positive cells and ROS-positive cells increase. Survival rate of the cells from aged rats is lower than that of the cells from young rats. At four week after induction with BMP-2, expression of Nkx2.5, GATA-4, cTnT mRNA and Cx-43 mRNA of the cells of young group increase significantly. In adult and aged group, expression of the cardiac transcription factors and cardiac specific genes is lower than that of the cells in young group. In immunocytochemical staining, cTnT expression of the cells in young group is stronger after induction with BMP-2. As compared with that of the cells in young group, cTnT expression of the cells in aged group is weak after induction. Conclusion With increasing of age, MCSCs show senescent changes, including their abilities of proliferation, survival and differentiation toward cardiomyocytes decrease.
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Objective To compare the effects between Milrinone and Lrh-BNP for treating the peripartum cardiomyopathy heart failure.Methods After treated by routine measure,39 serious PPCM patients were randomized into two groups,with 20 to Milrinone group for the advanced therapy between 5 to 7 days,and 19 to Lrh-BNP group therapy 24 hours. Results The total efficient rate in Lrh-BNP group(95%) was more higher than that in Milrinone group(73.6%).The index of heart function such as LVEF,FS,E/A,SV,CO,CI,24 hours urine quantity,dyspnea could improve in both groups after the therapy(P
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Objective To analyse the radiological features and the diagnostic method of pulmonary alveolar proteinosis(PAP).Methods X-ray and CT manifestations of PAP in 37 cases confirmed with fiberoptic bronchoscopy and bronchoalveolar lavage were studied retrospectively.Results The radiologic features of PAP could be characterized as geographic,the “crazy-paving” pattern,lobar or segmental consolidation(air-brochogram sign)and like intersititial fibrosis.The radiologic manifestations were stable and more serious than the symptoms.Conclusion PAP is of typical radiologic feature,it is not difficult for diagnosis in combination with clinical characteristics.