This study investigates the alteration in function and number of circulating endothelial progenitor cells (EPCs) in patients with aortic dissection (AD), compared with hypertensive patients, and its possible mechanism. Thirty-four patients with acute aortic dissection (AAD) and 20 patients with primary hypertension were involved. Flow cytometry analysis was performed to detect the number of CD34+/KDR+ cells, and acetylated low density lipoprotein (ac-LDL) and lectin fluorescent staining method was applied to test the number of cultured EPCs. In addition, EPC migration and proliferation were measured, and plasma interleukin 6 (IL-6) and interleukin 17 (IL-17) levels were investigated. The number of circulating EPCs in the AAD group was lower than that in the non-AD group, and the proliferation and migration of circulating EPCs in the AAD group were lower than that in the non-AD group. In addition, the number, proliferation, and migration of circulating EPCs were significantly inversely correlated with the aortic dissection detection risk score (ADD-RS). More importantly, increased plasma IL-6 and IL-17 level was found in the AAD group, and the two inflammatory factors were inversely associated with the function and number of circulating EPCs in the AAD group. We first demonstrated that the number and function of circulating EPCs are reduced in the AAD group, which may be partly related to upregulated plasma IL-6 and IL-17. Our study provides novel insight on the underlying mechanism and potential therapeutic target of AAD.

Background Acute myeloid leukemia (AML) is a common hematopoietic malignancy and have unsatisfactory prognosis. Our study aimed to identify hub genes in AML and explore potential biomarkers through integrated bioinformatics. Methods Microarray datasets were analyzed to screen the differentially expressed genes (DEGs). Functional enrichment analysis was performed, and protein-protein interaction (PPI) network was generated by the STRING (11.0) database and Cytoscape (3.7.2) software. Hub genes were screened and verified through GEPIA2 and GEO microarray database. Sensitivity of AML cell lines with high expression of hub genes to the small-molecule drugs were identified using GSCA Lite. Results A total of 456 DEGs were identified and top 100 genes were screened out, of which six genes (FLT3, PF4, CD163, MRC1, CSF2RB, PPBP) were upregulated in AML and individually had a worse prognosis by the overall survival (OS) analysis. AML cell lines with FLT3-overexpression and CSF2RB-overexpression were sensitive to most small-molecule drugs, while, AML cells with CD163-overexpression were only sensitive to a few drugs. However, sensitivity to Erlotinib was correlated with high expression of PF4 and PPBP. Conclusions In summary, FLT3, PF4, CD163, MRC1, CSF2RB, PPBP may be potential biomarkers and potential sensitive small-molecule drugs were correlated with overexpression of the biomarkers in AML.

目的:探讨双氢青蒿素(DHA)和达沙替尼(Das)联合用药对人肝癌细胞的抑制作用及其分子机制.方法:采用CCK8法、克隆形成实验、吖啶橙/溴化乙锭(AO/EB)染色法、流式细胞术分别进行细胞活性、细胞凋亡、细胞周期、线粒体膜电位、细胞内活性氧(ROS)含量进行检测评价双青蒿素和达沙替尼协同用药的药效,采用Western blot研究其作用机制.结果:(1)DHA和Das单独使用均抑制HepG2细胞的生长,两者在一定的浓度下联合发挥协同作用;联合用药结果显示细胞克隆数量少且集落小;(2)联合用药后,周期调控蛋白c-Myc、Cyclin E1和E2F1表达明显降低,细胞周期在G0/G1阻滞;(3)联合用药后细胞内ΔΨm明显下降且凋亡更加显著,抗凋亡蛋白Bcl-2和PARP表达明显减少,促凋亡蛋白Bim、Cleaved-Caspase 9和Cleaved-PARP的表达显著增加;(4)DHA应用可显著促进胞内ROS的产生,并且显著下调抗氧化蛋白Nrf2表达.结论:DHA和Das联合用药显著抑制HepG2的增殖活性,二者在一定浓度下发挥协同作用,促使周期阻滞,诱导细胞凋亡,分子机制与G1期周期阻滞及线粒体依赖的凋亡信号通路有关.

目的:探讨达沙替尼(dasatinib)体外对人肾癌细胞系786-O及769-P增殖、凋亡及迁移的影响及其分子机制.方法:不同浓度(0～2μmol/L)达沙替尼处理786-O细胞及769-P细胞(0μmol/L为空白对照组),采用MTT法检测细胞活力,划痕实验检测细胞迁移能力,Hoechst 33258染色观察细胞凋亡,流式细胞术检测细胞周期,Western blot法检测细胞周期和细胞凋亡相关蛋白的表达水平.结果:达沙替尼对786-O及769-P两种细胞均有活力抑制和迁移阻滞的作用,随着浓度增大抑制效果越显著(P<0.05).达沙替尼对786-O细胞和769-P细胞的IC50分别为(0.9587±0.0288)μmol/L和(0.7843±0.0660)μmol/L.达沙替尼处理24 h后,促凋亡蛋白cleaved caspase-3和cleaved caspase-9的表达显著增加(P<0.01),细胞周期蛋白D1的表达下降(P<0.05),细胞周期相关通路蛋白p53和p21表达量均显著增加(P<0.05),p-AKT蛋白水平显著下降(P<0.05).结论:达沙替尼通过上调p53表达并下调AKT磷酸化水平抑制人肾癌细胞系786-O及769-P的活力和迁移能力.

Background/Aims. Sexual differences exist in endothelial progenitor cells (EPCs), and various cardiovascular risk factors are associated with the preservation of endothelial function in premenopausal women. However, it is unclear whether differences in endothelial function and circulating EPCs exist between overweight premenopausal women and age-matched men. Methods. We compared EPC counting and functions in normal-weight and overweight premenopausal women and men, evaluated endothelial function in each group, and detected the expression of the guanosine triphosphate cyclohydrolase I (GTPCH I) pathway. Results. The number of EPCs was lower in the male group than in the female group, regardless of normal-weight or overweight status, and there was no significant difference between the different weight groups among females or males. Endothelial function and EPC migration and proliferation were preserved in overweight premenopausal women compared with overweight men as were nitric oxide (NO) levels in plasma and secreted by EPCs. Endothelial function, the circulating EPC population, and NO levels were not different between normal-weight and overweight premenopausal women. Flow-mediated dilatation was significantly correlated with EPC function, plasma NO levels, and EPC-secreted NO. Conclusions. This investigation provides the first evidence for sex-based differences in EPC activity and endothelial function in overweight middle-aged individuals; these differences are associated with alterations in NO production and may partly occur through downregulation of the GTPCH I pathway. The present results provide new insights into the mechanism underlying the preserved endothelial function in overweight premenopausal women and may uncover a potential therapeutic target for endothelial repair in overweight population.

目的 研究克拉屈滨(2-CdA)对内皮细胞活力、迁移、周期和分泌活性的影响,以评估2-CdA在l临床应用中对心血管的影响.方法 CCK-8法检测不同浓度2-CdA对内皮细胞生长的影响;划痕实验检测细胞迁移;流式细胞术检测细胞周期;吖啶橙/溴化乙啶法检测细胞凋亡;Gries法测定一氧化氮(NO)含量;ELISA法检测血管内皮生长因子(VEGF)水平.结果 在0.4～40 μmol/L浓度范围,2-CdA对内皮细胞有明显抑制作用,IC50为4.126μmol/L.随药物浓度升高和作用时间的延长,2-CdA的抑制作用增强;2-CdA高浓度长时间作用可导致内皮细胞凋亡.5 μmol/L 2-CdA作用内皮细胞后,细胞迁移能力受抑制,细胞周期明显阻滞在S期.吖啶橙/溴化乙啶细胞染色结果显示,高浓度2-CdA可诱导内皮细胞的凋亡.5μmol/L 2-CdA作用内皮细胞后导致NO和VEGF含量减少.结论 2-CdA具有内皮细胞毒性作用,引起细胞周期阻滞,并导致细胞凋亡.

AIM: To study the effects of cladribine on growth and secretion activity of human umbilical vein endothelial cell line EA.hy926, and to investigate the mechanism of its anti-tumor effect by inhibiting endothelial cells. METHODS:The effects of cladribine at different concentrations on the cell viability were detected by CCK -8 assay.Apop-tosis and cell cycle distribution were examined by flow cytometry.The protein expression levels were determined by Western blot.The levels of tumor necrosis factor-α(TNF-α), transforming growth factor-β1(TGF-β1)and vascular endothelial growth factor(VEGF)secreted by EA.hy926 cells with cladribine treatment for 48 h were analyzed by ELISA.The nitric oxide(NO)production was measured by Gries method.RESULTS:Cladribine at 0.4~1 μmol/L inhibited the viability of EA.hy926 cells in time-and dose-dependent manners.The IC50was about 3.644 μmol/L.The results showed 43.74% cells in S phase when the concentration of cladribine was 0.4 μmol/L,and 77.23 % cells in S phase when the concentra-tion of cladribine was 1 μmol/L.The apoptosis was not induced by cladribine at 0.4~10 μmol/L.The protein expression of Bax and caspase-3 did not change.The expression of p21 increased and the p53 decreased(P<0.05).The levels of TNF-αand TGF-β1 secreted by EA.hy926 cells increased after cladribine treatment for 48 h.The levels of VEGF and NO decreased.CONCLUSION:Cladribine obviously inhibits the viability of EA.hy926 cells.The mechanism is related to the cell cycle arrest.Cladribine promotes the secretion of TNF-αand TGF-β1 by EA.hy926 cells and inhibits the secretion of VEGF and NO.

Objective To screen the effective small interfering RNAs (siRNAs) targeting against long non-coding RNA (lncRNA) PVT1 (plasmacytoma variant translocation 1), and to observe its influence on the proliferation of human leukemic cell K562.Methods Four siRNA sequences targeting against PVT1 (PVT1-siRNA1, 2, 3, 4) and scramble control sequence (SC-siRNA) were designed and synthesized.Using HiperFect transfection reagent, all of the siRNAs were transfected into K562 cells.The experiment was divided into siRNA1 group, siRNA2 group, siRNA3 group, siRNA4 group, SC-siRNA group, idling group and untransfected group.The PVT1 RNA expression level was assessed by real-time RT-PCR.The cell proliferation was determined by CCK8 assay and the cell cycle was detected by Flow cytometry.Results The expression level of PVT1 RNA of K562 cells in siRNA4 group was significantly reduced compared with SC-siRNA group, idling group and untransfected group 24 and 48 hours after transfection (P<0.01), while there was no significant difference among siRNA1 group, siRNA2 group, siRNA3 group and untransfected group (P>0.05).The cell proliferation of K562 cells in the siRNA4 group was significantly inhibited as compared with SC-siRNA group and idling group 24, 48 and 72 hours after transfection (P<0.01).The cells in the siRNA4 group were arrested at G1 phase 48 and 72 hours after transfection (P<0.01).Conclusion PVT1-siRNA can inhibit the proliferation of K562 cells through arresting the cell cycle at G1 phase.

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.

Aim To explore the effect of dasatinib on proliferation,migration,cell cycle and apoptosis of human umbilical vein endothelial cells cytologically and molecularly to offer help on clinical application of dasatinib.Methods Experimental grouping:dasatinib group (dasatinib concentration was 50 nmol/L) and LY294002 group (PI3K inhibitor,concentration was 20 μmol/L),combined treatment group (dasatinib treatment concentration was 50 nmol/L,the concentration of LY294002 for 20 μmol/L) and mock group (DMSO was 0.1％).Cell viability was measured by CCK8.The migration ability of human umbilical vein endothelial cells was measured by scratch assay.Apoptosis and cell cycle were analyzed by flow cytometry.The expression and phosphorylation of Akt protein were determined by Western blot.Results The cell viability of human umbilical vein endothelial cells decreased gradually following the increasing concentration (1 ～ 400 nmol/L) and prolonged exposure to dasatinib (50 nmol/L,24 ～ 96 h).Dasatinib and LY294002 (an inhibitor of PI3K),both inhibited cell viability of human umbilical vein endothelial cells.Dasatinib (50～ 100 nmol/L) weakened the migration capability of human umbilical vein endothelial cells.In addition,compared with control,dasatinib (50 nmol/L) induced apoptosis and cell cycle arrest (P＜0.05) of human umbilical vein endothelial cells.The phosphorylation of Akt was inhibited by dasatinib and LY294002,and LY294002 had more powerful inhibition of p-Akt than dasatinib.Conclusion Dasatinib could facilitate the injures of human umbilical vein endothelial cells via PI3K/Akt pathway,mainly to suppress the proliferation and migration of human umbilical vein endothelial cells,alter the cell morphology,blocked the G1-S transition and induced apoptosis.

Objective To study the influence of down-regulating long non-coding RNA(lncRNA)plasmacytoma variant translocation 1(PVT1) andMYC-induced long noncoding RNA(MINCR)on the proliferation of lymphoma cells Raji and the probable mechanism.Methods Small interfering RNA(siRNA)targeting against PVT1 and MINCR as well as scramble control siRNA(SC-siRNA)were designed and synthesized chemically.The experiment was divided into PVT1-siRNA group, MINCR-siRNA group, PVT1-siRNA+MINCR-siRNA group, scramble control group and untransfected cells group.Using HiperFect transfection reagent,all of the siRNAs were transfected into Raji cells.The MINCR RNA expression level was detected by real-time RT-PCR.The cell proliferation, the cell cycle and the expression of c-Myc protein were detected by CCK8 assay, Flow cytometry and Western blot, respectively.Results The expression level of MINCR RNA ofMINCR-siRNA group was significantly reduced compared with that of the scramble control group and untransfected cells group(P<0.05).The cell proliferation was significantly inhibited after PVT1-siRNA combining with MINCR-siRNA transfected into Raji cells (P<0.05),which was more obvious than that in PVT1-siRNA group and MINCR-siRNA group(P<0.05).Consistently,the cells were arrested in G1 phase after PVT1-siRNA combining with MINCR-siRNA transfected into Raji cells (P<0.05),and it was more obvious compared with PVT1-siRNA group and MINCR-siRNA group(P<0.05).The expression of c-Myc protein was significantly decreased after PVT1-siRNA combining with MINCR-siRNA transfected into Raji cells(P<0.05),which was more obvious compared with PVT1-siRNA group and MINCR-siRNA group(P<0.05).Conclusion Down-regulating PVT1 and MINCR can enhance the inhibition of proliferation of Raji cells.

AIM:To explore the protective effects of salidroside on endothelial progenitor cells (EPCs) dam-aged by radiation and its mechanisms.METHODS:EPCs from normal peripheral blood were cultured in fibronectin-coated flasks with endothelial progenitor medium.The effects of salidroside on the viability, migration, adhesion and apoptosis of radiation-damaged EPCs were detected.The viability, apoptosis and migration of the cells were assayed by CCK-8 assay, flow cytometry and Transwell chamber experiment, respectively.The cell adhesion assay was performed by re-plating the cells on fibronectin-coated dishes, and then the adherent cells were counted.The expression of Akt protein in the cells was assessed by Western blotting.RESULTS:Salidroside improved the viability, and migratory and adhesive capacities of the EPCs, and decreased the apoptosis after radiation.Salidroside also increased the protein level of phosphorylated Akt.How-ever, the effects of salidroside on radiation-damaged EPCs were inhibited by phosphatidylinositol 3-kinase inhibitor LY294002.CONCLUSION: Salidroside protects EPCs from radiation damages and its mechanism is associated with en-hancing phosphatidylinositol 3-kinase/Akt signaling pathway.

AIM:Toinvestigatetheeffectofnicotinicacidamide(NAA)ontheinfusiondamageofhuman umbilical cord mesenchymal stem cells ( hUC-MSCs) under the condition of instant blood-mediated inflammatory reaction ( IBMIR) .METHODS:Normal peripheral blood without anticoagulant at volume of 2.7 mL was mixed with 0.3 mL phys-iological saline (as blank group), CFSE labeled hUC-MSCs (1 ×106 cells in 0.3 mL as MSC group) and CFSE labeled hUC-MSCs (1 ×106 cells in 0.3 mL) preprocessed with NAA at concentration of 10 mmol/L for 24 h ( as MSC+NAA group) , respectively.The mixture was immediately injected into the improved Chandler Loop model, placed in 37℃water bath, and then started the peristaltic pump at the speed of 20 mL/min for 1 h.The number of CFSE labeled hUC-MSCs, platelets, white blood cells were counted and the concentration of complement C3a was measured before and after cycling, respectively.RESULTS: After 1 h circulation, the platelet dissipation rate were ( 29.96 ±10.88 )% in blank group, (77.76 ±19.29)% in MSC group all and (50.13 ±18.10)% in MSC +NAA group; and the leukocyte counts were (37.82 ±13.81)%in blank group, (64.57 ±17.08)% in MSC group and (41.52 ±17.26)% in MSC+NAA group. Compared with blank group, the differences of the dissipation rates in MSC group and MSC+NAA group all had statistical significance.The hUC-MSCs relative survival rate in MSC+NAA group was higher than that in MSC group.C3a concentra-tions in blank group, MSC group and MSC+NAA group were (206.27 ±58.10), (230.47 ±39.61) and (208.37 ± 40.66) μg/L, respectively.CONCLUSION:Co-circulating the mixture of hUC-MSCs with normal peripheral blood with-out anticoagulant in the improved Chandler Loop for 1 h depletes a large number of hUC-MSCs and blood components, and increases C3a, suggesting that this model can induce IBMIR.NAA has a protective effect on the hUC-MSCs in the infusion damage by inhibiting IBMIR, reducing the wastage of the blood components and enhancing the survival rate of the hUC-MSCs.

Cell-based therapies are major focus of current research for treatment of liver diseases. In this study, mesenchymal stem cells were isolated from human umbilical cord Wharton's jelly (WJ-MSCs). Results confirmed that WJ-MSCs isolated in this study could express the typical MSC-specific markers and be induced to differentiate into adipocytes, osteoblasts, and chondrocytes. They could also be induced to differentiate into hepatocyte-like cells. Poly (3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate) (PHBVHHx) is a new member of polyhydroxyalkanoate family and biodegradable polyester produced by bacteria. PHBVHHx scaffolds showed much higher cell attachment and viability than the other polymers tested. PHBVHHx scaffolds loaded with WJ-MSCs were transplanted into liver-injured mice. Liver morphology improved after 30 days of transplantation and looked similar to normal liver. Concentrations of serum alanine aminotransferase and total bilirubin were significantly lower, and albumin was significantly higher on days 14 and 30 in the WJ-MSCs+scaffold group than in the carbon tetrachloride (CCl4) group. Hematoxylin-eosin staining showed that liver had similar structure of normal liver lobules and similar size and shape of normal hepatic cells, and Masson staining demonstrated that liver had less blue staining for collagen after 30 days of transplantation. Real-time reverse transcription-polymerase chain reaction (RT-PCR) showed that the expression of the bile duct epithelial cell gene CK-19 in mouse liver is significantly lower on days 14 and 30 in the WJ-MSCs+scaffold group than in the CCl4 group. Real-time RT-PCR, immunocytochemistry, and periodic acid-Schiff staining showed that WJ-MSCs in scaffolds differentiated into hepatocyte-like cells on days 14 and 30 in the WJ-MSCs+scaffold group. Real-time RT-PCR also demonstrated that WJ-MSCs in scaffolds expressed endothelial cell genes Flk-1, vWF, and VE-cadherin on days 14 and 30 in the WJ-MSCs+scaffold group, indicating that WJ-MSCs also differentiated into endothelial-like cells. These results demonstrated that PHBVHHx scaffolds loaded with WJ-MSCs significantly promoted the recovery of injured liver and could be further studied for liver tissue engineering.

Current therapy for skin wound healing still relies on skin transplantation. Many studies were done to try to find out ways to replace skin transplantation, but there is still no effective alternative therapy. In this study, decellularized scaffolds were prepared from pig peritoneum by a series of physical and chemical treatments, and scaffolds loaded with hyaluronic acid (HA) and epidermal growth factor (EGF) were tested for their effect on wound healing. MTT assay showed that EGF increased NIH3T3 cell viability and confirmed that EGF used in this study was biologically active in vitro. Scanning electron microscope (SEM) showed that HA stably attached to scaffolds even after soaking in PBS for 48h. ELISA assay showed that HA increased the adsorption of EGF to scaffolds and sustained the release of EGF from scaffolds. Animal study showed that the wounds covered with scaffolds containing HA and EGF recovered best among all 4 groups and had wound healing rates of 49.86%, 70.94% and 87.41% respectively for days 10, 15 and 20 post-surgery compared to scaffolds alone with wound healing rates of 29.26%, 42.80% and 70.14%. In addition, the wounds covered with scaffolds containing EGF alone were smaller than no EGF scaffolds on days 10, 15 and 20 post-surgery. Hematoxylin–Eosin (HE) staining confirmed these results by showing that on days 10, 15 and 20 post-surgery, the thicker epidermis and dermis layers were observed in the wounds covered with scaffolds containing HA and EGF than scaffolds alone. In addition, the thicker epidermis and dermis layers were also observed in the wounds covered with scaffolds containing EGF than scaffolds alone. Skin appendages were observed on day 20 only in the wound covered with scaffolds containing HA and EGF. These results demonstrate that the scaffolds containing HA and EGF can enhance wound healing.

Objective To investigate the potential and underlying molecular mechanism of salidroside in ameliorating radiation-induced bone marrow adipogenesis and stimulating hematopoiesis.Methods The female BALB/c mice aged 6-7 weeks were randomly divided into normal control group,radiation group and salidroside group.The radiation group and salidroside group were irradiated with 6.0 Gy of 60Co γ-rays.The salidroside group was intraperitoneally injected with 30 mg· kg-1 · d-1 salidroside at 12 h and then every day until 8th d after radiation.The normal control group and radiation group were treated with equal volume of saline as control of salidroside.At 14 d after radiation,the mice weight,peripheral blood count,femur bone marrow histology,and the proportion of adipocyte area were measured,and the expressions of PPAR-γ and FABP4 were detected by q-PCR.Results After irradiation,the numbers of white blood cells,hemoglobin and platelet in peripheral blood were reduced obviously,and the percentage of adipocyte area was increased significantly.Compared with mice in the radiation group,salidroside inhibited adipogenesis and reduced the proportion of adipocyte area (t =13.31,P ＜ 0.05) by reducing the expressions of PPAR-γ and FABP4 (t =8.64,13.19,P ＜ 0.05).The number of white blood cells was partly recovered at 7 d after irradiation (t =5.80,P ＜ 0.05).Both white blood cells and hemoglobinin in peripheral blood of the salidroside group were higher than those in the radiation group at 14 d after irradiation.Conclusions Salidroside could inhibit radiation-induced bone marrow adipogenesis and regulate bone marrow microenvironment,thereby promotes hematopoietic recovery in mice after radiation injury.

Objective To construct an FABP4 promoter recombined luciferase reporter gene vector.and to detect its function,thus providing a basis for the research on lipid metabolism related medicines.Methods Primers were designed based on human FABP4 gene promoters,and FABP4promoters from human genome DNA was replicated by PCR.Then FABP4 promoters were inserted into the pGL3-basic vector after double digestion by restriction enzyme KpnI and XhoI.Positive clones were identified by sequencing.The recombinant pGL3-basic-FABP4-promoter vector were transiently transfected into K293 cells.After 24 hours,the activity of luciferase was detected.Results A pGL3 luciferase reporter vector was constructed.The result of sequencing and double digesting of recombined plasmid were completely correct.The results of luciferase detection after transfection showed that the transcriptional activity of recombinant pGL3 plasmid was obviously increased compared with that of pGL3- basic plasmid(P0.001).Conclusion A pGL3 luciferase reporter vector containing human FABP4 promoter gene was constructed successfully,which provides a tool and basis for further study of FABP4 gene expression regulation.

Objective To investigate the effect of mouse fetus-derived mesenchymal stem cells ( MSCs) transplantation on lung func-tion in aged mice.Methods The 12-month-old BALB/c mice were randomly divided into control and transplantation groups .MSCs were de-rived from 13.5 d fetuses.The mice in transplantation group were injected with fetus-derived MSCs through the tail vein , while mice in con-trol group were injected with equal volume of normal saline .Morphologic analysis of the lung tissue was performed .The pulmonary artery blood flow velocity and cardiac function were measured by echocardiography .Results There were the senile emphysema changes in control group, MSCs significantly ameliorated the senile emphysema changes in transplanted mice .Pulmonary artery blood flow velocity in transplan-tation group was slow down , and the stroke volume , cardiac output and left ventricular diastolic volume were increased .Weight-loaded swimming time of MSCs transplantation group was longer , blood lactic acid levels before and after swimming were lower than those in control group.Conclusions Transplantation of mouse fetus-derived MSCs could delay degenerative changes of the lung , improve the lung function of the aged mice .

[ABSTRACT]AIM:ToinvestigatetheeffectofmiR-155-specificsiRNAaloneorincombinationwithcytosinear-abinoside (Ara-C) on the growth and apoptosis of Burkitt lymphoma Raji cells .METHODS: miR-155-specific siRNA and/or Ara-C were used to treat the cells .Quantitative real-time polymerase chain reaction was used to detect the expres-sion of miR-155.The growth of the cells was analyzed by CKK-8 assay.The cell apoptosis was determined by flow cytome-try.RESULTS:The miR-155 expression level of the cells transfected with miR-155 siRNA was significantly lower than that in the 2 control groups .Ara-C or miR-155 siRNA alone inhibited the growth of Raji cells in a dose-depend manner . miR-155 siRNA combined with Ara-C produced more inhibition of cell proliferation (P<0.05).After treatment for 48 h, the apoptotic rate of Raji cells in miR-155 siRNA+Ara-C group [(38.4 ±1.4)%] was higher than that in Ara-C group [(16.5 ±0.3)%] and miR-155 siRNA group [(14.6 ±0.3)%], with statistically significant difference (P<0.05). The expression of caspase-3 in Ara-C+miR-155 siRNA group was increased significantly as compared with Ara-C group and miR-155 siRNA group.CONCLUSION:miR-155-specific siRNA enhances the chemosensitivity of Raji cells to Ara-C by inducing apoptosis through the caspase-3 pathway .

目的:探讨光敏化姜黄素对人胃腺癌MGC-803细胞的影响及其细胞内的分布情况.方法:用MTT法检测细胞株的增殖抑制率,流式细胞仪检测细胞的凋亡率,荧光显微镜观察细胞核形态;激光共聚焦显微镜观察姜黄素在细胞内的分布.结果:光敏化姜黄素作用于MGC-803细胞12、24、48 h后,MTT法检测细胞的抑制率分别为35.53％±3.52％、40.17％±2.01％、44.93％±3.61％;凋亡率分别为20.32％±3.07％、22.46％±1.51％、26.58％±2.67％,与对照组比较差异有统计学意义(P＜0.05).光敏化姜黄素作用48 h后能见明显的凋亡小体.激光共聚焦显微镜下观察光敏化姜黄素发出的黄色荧光主要在细胞质膜、核膜呈聚集性分布,散在分布于其他亚细胞结构,细胞核分布较少.结论:光敏化姜黄素能抑制胃癌细胞的生长及凋亡,具有显著的光动力学效应,随作用时间的延长而逐渐增加.光敏剂主要分布于细胞质膜和核膜,散在分布于其他亚细胞结构,细胞核分布较少.