Effects of resveratrol on aging of mesenchymal stem cells and its mechanism / 浙江大学学报·医学版

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.

Sedentary behaviors and physical fitness among middle school students / 中国学校卫生

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.

Derivative of Stevioside; CPUK02; Restores ESR1 Gene Methylation in MDA-MB 231

Background: CPUK02 (15-Oxosteviol benzyl ester) is a new ent-kaurenoid derivative of stevioside and exhibits strong anti-cancer activity. Nowadays, the pattern of epigenetic in cancer has been topic of many studies and DNA methylation targeting represents a relevant strategy for cancer treatment. Since, no study conducted to this mechanism, we attempt to evaluate whether CPUK02 induce its anti-cancer effects via alteration the level of mRNA DNMT3B, DNMT3A expression and ESR1 methylation pattern in breast cancer cells line. Methods: MCF-7 (ER +) and MDA-MB231 (ER-) cell lines were treated for 24, 48 hours with 1 µM CPUK02 and 5-AZA-CdR (DNA methyltransferase inhibitor). Quantitative expression of DNMT3B and DNMT3A genes and ESR1 promoter methylation was assessed by Real-Time PCR and MS-PCR, respectively. Results: CPUK02 restored ESR1 promoter unmethylated allele in MDA-MB 231 cells. Also treatment with CPUK02 decreased the expression of both DNMT3A and DNMT3B genes like 5-AZA. The expression of DNMT genes were diminished by half compared with control cells. Conclusions: These results showed that CPUK02 has an anticancer effect on MDA-MB 231 cells which this effect can be done through several pathways.

Salinity Tolerance Mechanism of Osmotin and Osmotin-like Proteins: A Promising Candidate for Enhancing Plant Salt Tolerance.

INTRODUCTION: Salt stress is one of the most important abiotic stress factors which severely affect agricultural production. Osmotins and OLPs (osmotin like proteins) are kinds of proteins which were produced during plant adapting to the environmental stress. OBJECTIVE: These proteins were closely related to osmotic regulation and resistance stress. They are widely distributed in plants. Their expression for these genes was induced by salt stress, which played important roles in plants responding to salt stress. CONCLUSION: During salt stress, osmotin can help accumulate proline, and quench reactive oxygen species and free radicals.

Induction of Epigenetic Alteration by CPUK02, An Ent- kaurenoid Derivative of Stevioside.

BACKGROUND: Dietary polyphenols, such as those found in green tea and red wine, are linked to antitumor activity. They are known to influence many signaling pathways epigenetically within the human body. In this regard, CPUK02 (15-Oxosteviol benzyl ester) is a new ent-kaurenoid derivative of stevioside and exhibits strong anti-cancer activity in vitro and in vivo. Nowadays, the role of epigenetics in cancer has been the subject of intensive study and DNA methylation targeting represents a relevant strategy for cancer treatment. There are no reports regarding the effects of CPUK02 on epigenetic alterations in colorectal cancer cell line. This study was an attempt to compare CPUK02 with 5-AZA as DNMT inhibitor agent and evaluate whether it can induce its anti-cancer effects via altering the level of DNMT3b mRNA, MGMT and SFRP2 methylation pattern in HCT 116 cell line. METHODS: To evaluate DNMT3b expression, DNMT3B mRNA levels in HCT116 CRC cell line were quantified by real-time reverse-transcriptase Polymerase Chain Reaction (PCR) assay after 24 hr of incubation time with CPUK02 and 5-AZA. In addition, the methylation patterns of 2 CpG islands in this cell line were examined by methylation-specific PCR methods. RESULTS: CPUK02 surprisingly, decreased the DNMT3b mRNA level. The average expression levels of DNMT3b in HCT116 treated with CPUK02 and 5-AZA relative to the GAPDH expression level in control were 0.16 and 0.5%, respectively. Furthermore, CPUK02 could decrease the methylated allele of MGMT and SFRP2 genes in HCT 116 after 24 hr. CONCLUSION: In this study, positive correlation was found between mRNA expression of DNMT3b and gene promoter hypermethylation after treatment with CPUK02 and 5-AZA. Our data confirmed that CPUK02 like 5-AZA exhibits demethylating properties.

A new synthetic process of dapagliflozin / 中国药科大学学报

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.

Assessment of effect of laboratory external quality assessment program on cytogenetic testing / 重庆医学

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 .

The expression changes of peripheral endothelial progenitor cells and Ang-1/Tie2 in patients with pulmonary arterial hypertension / 中华检验医学杂志

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.

Effects of high Annexin A3 expression on apoptosis and proliferation of gastric cancer MGC-803 cells / 临床与实验病理学杂志

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.

Reviews on Historical Evolution and Modern Research of Rhizoma Pinelliae Fermentata / 世界科学技术-中医药现代化

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.

Morphology effect on the light scattering and dynamic response of polymer network liquid crystal phase modulator.

Polymer network liquid crystal (PNLC) was one of the most potential liquid crystal for submillisecond response phase modulation, which was possible to be applied in submillisecond response phase only spatial light modulator. But until now the light scattering when liquid crystal director was reoriented by external electric field limited its phase modulation application. Dynamic response of phase change when high voltage was applied was also not elucidated. The mechanism that determines the light scattering was studied by analyzing the polymer network morphology by SEM method. Samples were prepared by varying the polymerization temperature, UV curing intensity and polymerization time. The morphology effect on the dynamic response of phase change was studied, in which high voltage was usually applied and electro-striction effect was often induced. The experimental results indicate that the polymer network morphology was mainly characterized by cross linked single fibrils, cross linked fibril bundles or even both. Although the formation of fibril bundle usually induced large light scattering, such a polymer network could endure higher voltage. In contrast, although the formation of cross linked single fibrils induced small light scattering, such a polymer network cannot endure higher voltage. There is a tradeoff between the light scattering and high voltage endurance. The electro-optical properties such as threshold voltage and response time were taken to verify our conclusion. For future application, the monomer molecular structure, the liquid crystal solvent and the polymerization conditions should be optimized to generate optimal polymer network morphology.

Effects of D-galactose on ageing of rat mesenchymal stem cells / 浙江大学学报·医学版

<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>

Research progress of the biological characteristics of IkappaB kinase and its inhibitors / 药学学报

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.

Synthesis and antitumor activity of ent-kaurene diterpenoids / 中国药科大学学报

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.

Comparative study on biological characteristics of marrow-derived cardiac stem cells from rats at different Ages / 解剖学报

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+G2/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.

Effects of Nitroglycerin and Lyophilized Recombinant Human BNP on Treatment of the Peripartum Cardiomyopathy Heart Failure / 医学研究杂志

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

Analysis of Radiological Features of Pulmonary Alveolar Proteinosis / 实用放射学杂志

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.