Objective To explore the inhibitory effect of astragalus polysaccharide (APS) on the proliferation of human erythroleukemia K562 cells and its mechanisms.Methods After K562 cells (purchased from Shanghai cell bank of chinese academy of science) were treated with different concentrations (0 mg/L,100 mg/L,200 mg/L and 400 mg/L) of APS.The influences of APS on the growth rate,doubling time and cell cycle distribution of K562 cells were observed by methyl thiazolyl tetra-zolium assay (MTF) and flow cytometry,respectively.Furthermore,the reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting assay were used to detect the expressions of Cyclin A,Cyclin B,Cyclin E and p21 gene at the mRNA and protein levels,respectively.Results MTT assay findings showed that,compared to the control group (0 mg/L APS),growth rates of K562 cells treated with 100 mg/L,200 mg/L and 400 mg/L APS decreased significantly (all P ＜ 0.01),and the doubling times lengthened significantly (all P ＜ 0.01).Flow cytometry findings revealed that,compared to the control group,the G1 phase cells in K562 cells of APS group increased significantly (P ＜0.01),while the S and G2/M phase cells decreased significantly (all P ＜ 0.01).RT-PCR and Western blotting results indicated that Cyclin B and Cyclin E expression of K562 cells at the mRNA and protein levels in the APS group were significantly lower than those of the control group(all P ＜ 0.01),whereas p21 expression was significantly enhanced at mRNA and protein levels (P ＜ 0.01),and Cyclin A expression was not significantly different at mRNA and protein levels between the 2 groups (all P ＞ 0.05).Conclusions APS could inhibit the proliferation of human erythroleukemia K562 cells.APS could inhibit the proliferation of K562 cells by down-regulating the expression of Cyclin B and Cyclin E and up-regulating the expression of p21.

BACKGROUND:Previous studies have demonstrated that transplanted neural stem cells can survive and proliferate in the brain of Alzheimer disease(AD)rats,however,it is poorly understood whether it can rebuild the nerve tracts by substituting the injured or dead neurons and improve learning and memory abilities.Synaptophysin is one of the important markers of synaptic rebuilding.OBJECTIVE:To observe the effects of neural stem cell transplantation on synaptophysin expression in hippocampus and learning and memory abilities of AD rats.METHODS:Sprague Dawley rats were randomly divided into the normal control,AD model,2-week-transplantation and 4-week-transplantation groups.All rats were established AD models except that in the normal control group.Neural stem cells were isolated from the dentate gyrus of hippocampus of newborn rats,labeled with Hoechst33258,and then transplanted into CA1 region of hippocampus of rats in the 2-week-transplantation and 4-week-transplantation groups.The behavioral testing in the rats was performed using Y-maze trial.Nissl staining and synaptophysin immunohistochemistry were detected after the rats were sacrificed.The same volume of stroke-physiological saline solution was injected into rats in the AD models group using the identical methods.There was no treatment in the normal control group.RESULTS AND CONCLUSION:①The cells number in the hippocampal CA1 region of the 2-week-transplantation and 4-week-transplantation groups were increased than that of AD model group,but were still less than that of the normal control group(P ＜ 0.05).There was no significantly difference between the absorbance values of 2-or 4-week-transplantation group and control group(P ＞ 0.05).②The absorbance values of the 2-week-transplantation and 4-week-transplantation were significantly greater than that of the control and AD model groups(P ＜ 0.05).③The learning and memory abilities in 2-and 4-week-transplantation group enhanced obviously and their correct reaction rates improved evidently,which was found statistically significant difference from AD model group(P ＜ 0.05),while no statistically significant difference from control group(P ＞ 0.05).The transplanted neural stem cells may promote the synaptic rebuilding and improve learning and memory abilities in AD rats.

Objective Using the evidence-based medicine to investigate the effect of cognitive behavior intervention on physical and mental health in patients with type 2 diabetes mellitus.Methods Controlled studies related to psychological intervention on depression and glyeosylated hemoglobin in patients with type 2 diabetes were retrieved from electronic databases such as CNKI,VIP,Wanfang database and Pubmed,etc.The quality of included studies was evaluated and then data were analyzed by using RevMan5 software.Results A total of 273 articles were retrieved and finally 6 were enrolled.Results of Meta analysis were listed as follows:Cognitive behavior intervention could reduce score of Self-rated Depression Scale (SDS) in patients with diabetes mellitus.Cognitive behavior intervention could effectively reduce glycosylated hemoglobin values in patients with diabetes mellitus.Conclusions Cognitive behavior intervention can improve physical and mental health,decrease glycosylated hemoglobin and improve depression status in patients with type 2 diabetes mellitus.However,all the trials included in this review are of low quality; larger scale RCTs of higher quality are needed to confirm this conclusion.

Objective To observe the effects of tetra-atsenic oxide (As4O6) on the proliferation, migration and invasion of human breast cancer MCF-7 cells; To explore its potential mechanism. Methods The human breast cancer MCF-7 cells were regarded as the research object and cultured in vitro, with different concentrations of As4O6 using to intervene in MCF-7 cells. The cell proliferation was detected by MTT assay; the flow cytometry and wound-healing assay were used to detect the cell apoptosis and migration, respectively. The expressions of Cyclin E1, Cyclin A2, Caspase 3, p21 and MMP-9 mRNA were accessed by semi quantitative RT-PCR. Results As4O6 had a significant inhibitory effect on the proliferation of MCF-7 cells in a dose dependent manner. Compared with the control group (0 μmol/L), the apoptosis rate increased significantly when the concentration of As4O6 was 9, 12, 15 μmol/L (P<0.01). Either As4O6 at high (3 μmol/L) or low (1 μmol/L) concentration could effectively inhibit the migration of MCF-7 cells (P<0.01). With the increasing concentration of As4O6, the expressions of Cyclin E1, Caspase 3, and p21 mRNA significantly increased, while the expressions of Cyclin A2 and MMP-9 mRNA significantly decreased (P<0.05, P<0.01). Conclusion As4O6 can significantly inhibit the proliferation, cycle, invasion and migration of breast cancer MCF-7 cells, and the mechanism may be related to the increase of expressions of cyclin E1, caspase 3, p21 and inhibition of expressions of cyclin A2 and MMP-9.

AIM: To study the effect of monocyte/macrophages treated with CpG-oligodeoxynucleotides on leukemic K562 cells. METHODS: The monocytes/macrophages from peripheral blood cells were isolated and induced. The expressions of CD14 and CD16 on monocytes/macrophages were detected by means of flow cytometry. After treated with synthetic CpG-oligodeoxynucleotides, and nonCpG-oligodeoxynucleotides for 24 hours respectively, the inhibiting effect of monocyte/macrophages on K562 cells were detected using MTT method. The secretions of TNF-? and IL-12 from monocytes/macrophages were determined using ELISA method. RESULTS: The monocytes/macrophages treated with CpG-oligodeoxynucleotides enhanced their antitumor effect on K562 cells and increased the secretion levels of TNF-? and IL-12. Whereas, there was no significant difference between antitumor effect and cytokine secretion of the monocytes/macrophages treated with nonCpG-oligodeoxynucleotide. CONCLUSION: CpG-oligodeoxynucleotides increases the cytotoxicity of macrophages on K562 cells in vitro, as well as facilitates the IL-12 and TNF-? secretion. It provides a new approach for immunologic treatment of leukemia.

AIM:To explore the effect of dasatinib on the viability, migration, cell cycle and apoptosis of human bone marrow mesenchymal stem cells (hBMSCs), as well as the underlying signal pathway to evaluate the influence of dasatinib on hematopoietic microenvironment clinically.METHODS:The cell viability was measured by CCK-8 assay.The migration ability was detected by wound healing assay.The cell cycle and apoptosis were analyzed by flow cytometry.Acridine orange/ethidium bromide staining was also used to detected apoptosis.The secretion of transforming growth factor-β1 (TGF-β1) and tumor necrosis factor-α (TNF-α) were measured by ELISA.The protein levels of cleaved caspase-3, protein kinase B (Akt) and phosphorylated Akt were determined by Western blot.RESULTS:Compared with control group, dasatinib at 1~10 nmol/L suppressed the viability and migration ability of hBMSCs, and dasatinib at concentration of 7 nmol/L was adopted in the following assays.Dasatinib promoted apoptosis, and blocked the cell cycle in G1 phase.In addition, the secretion of TGF-β1 and TNF-α was increased markedly.The protein levels of cleaved caspase-3 was increased, but the protein levels of Akt and phosphorylated Akt were decreased.CONCLUSION:Dasatinib inhibits the viability and migration ability of hBMSCs in a dose-dependent manner, promotes the secretion TGF-β1 and TNF-α, and induces cell cycle arrest and apoptosis.Dasatinib might regulate the biological behaviors of hBMSCs observed above by modulating the expression and phosphorylation of Akt.

Objective To explore the potential role and function of miR-30 a in myocardial fibrosis after myocardi-al infarction( MI) .Methods We constructed the AAV plasmid vector which carried the miR-30 a gene of rat.The recombinant plasmid was detected by gene sequencing , enzyme digestion and PCR .Virus was packaged into HEK293 cells and virus titer was determined after extraction and purification by PCR .PBS fluid, rAAV9-miR-30 a-NC and rAAV9-miR-30 a were transmited to rat hearts from PBS group , miR-30 a-NC group and miR-30 a group respectively through transcoronary infusion before anterior descending coronary artery ligation .Sham group was set up at the same time.After 4 weeks, heart function was monitored by serial echocardiography , including fractional shortening ( FS) , and left ventricular ejection fraction ( LVEF) .Masson staining was used to calculate collagen volume fraction ( CVF) .The expression of collagen ⅠandⅢwere detected by immunohistochemistry . The mRNA level of miR-30a, TGF-β1 and CTGF were detected by real-time PCR.The protein level of TGF-β1 and CTGF were detected by western blot analysis .Results The cardiac function of miR-30 a group was improved significantly compared with PBS group and miR-30a-NC group (P<0.05).The levels of CVF,collagenⅠ,Ⅲexpression and Collagen Ⅰ/Ⅲ ratio in miR-30 a group was significantly lower than PBS group and miR-30 a-NC group ( P<0.01 ) .The mRNA and protein level of TGF-β1 and CTGF in miR-30 a group were reduced signifi-cantly than PBS group and miR-30 a-NC group ( P<0.001 ) .Conclusions The overexpression of miR-30 a after MI may reduce the mRNA and protein level of TGF-β1 and CTGF, so as to suppress myocardial fibrosis and im-prove cardiac function.

Humans ; Immunoglobulin A ; blood ; Immunoglobulin Light Chains ; blood ; Leukemia ; blood ; Multiple Myeloma ; blood

In order to guarantee the quality of traditional Chinese medicines (TCMs), the crystallization transformation of complex extracts of TCMs and the influence of solid form on their physicochemical properties were studied.The extract of total flavonoids from Pueraria lobata was taken as a model.Crystallization transformation happened when lofting under different conditions, and the intrinsic dissolution tests were carried out.It was found that humidity was the key factor to induce crystallization of total flavonoids from Pueraria lobata.The greater the wettability was, the more the crystallization was.The dissolution rate of total flavonoids from Pueraria lobata with the most crystallization amount significantly decreased by 96.51% compared to the sample without crystallization.After further simulating the preparation process of total flavonoids from Pueraria lobata, it was found that the wet granulation process with introduced water would also lead to crystallization and reduced dissolution rate.As for all crystallization samples, there was an inversely proportional relationship between the dissolution rates and the amount of crystallization.The risk of crystallization existed both in the storage and preparation process of TCM extracts.Crystallization would significantly affect the dissolution rate, and thus the quality of TCM products.In this study, the crystallization transformation of amorphous complex TCM extracts was discovered, and the effect of the crystallization transformation on its dissolution behavior was systematically studied, which provides a new research idea for assuring the quality of TCM products and promoting the improvement of TCM preparation level.