AIM: To investigate the effects of apelin-13 on proliferation, migration and capillary-like tube formation of a monkey choroid / retinal endothelial cell line, RF/6A, to clarify whether apelin-13 could promote retinal angiogenesis in vitro.METHODS: RF/6A cells in good conditions were administrated with DMSO (the control group), apelin-13 at 0.1μmol/L (low dose group) or apelin-13 at 1μmol/L (high dose group).Cell proliferation, migration and capillary-like tube formation were detected by using the MTT assay, scratch assay and matrigel assay, respectively, at 24h after plating the cells.RESULTS: Cell proliferation was promoted in both low and high dose apelin-13 groups compared to the control cells (P<0.05);the cell migration distance of both apelin-13 groups was significantly greater than that of the control group (P<0.05);and the number of capillary-like tube structures of both apelin-13 groups was significantly larger than that of the control cells (P<0.05).In addition, cell proliferation, migration and tube formation increased as the concentration of apelin-13 increased.CONCLUSION: Apelin-13 could obviously promote the angiogenesis capacity of RF/6A cells, suggesting that apelin-13 was an important pro-angiogenic factor in retinal endothelial cells.

Objective To investigate the quality of autoantibody testing around the whole country.Methods Laboratories that perform autoantibody testing were recruited by letters or telephone communications.The auto-antibodies examined by the quality-control survey included anti-nuclear antibody (ANA),anti-double-stranded DNA (anti-dsDNA) antibody,anti-extractable nuclear antigens (anti-ENA),antimitochondria antibody (AMA),anti-smooth muscle antibody (ASMA),and anti-citrulline antibody (CCP).Each autoantibody was tested for 3 samples,so 15 samples were tested in total.Sample distribution and data analysis were double-blinded.Qualitative interpretation,staining patterns were evaluated by IIF.The agreement with qualitative interpretation for each specimen was evaluated by ELISA,and Immuno-Blot/Dot-Blot.Results One hundred and eight laboratories participated in this study.The testing methods included indirect immumofluorescence (IIF),immuno-Blot (IB),Dot-Blot (DB),double diffusion method (DID),enzymelinked immunosorbent assay (ELISA),chemo-illuminescent assay,Dot-immunogold filtration assay.The accuracy rates were 82％,83％,95％,96％,86％,respectively for ANA,anti-dsDNA,AMA,ASMA,and anti-CCP antibody.Anti-ENA were further divided into anti-SNP,anti-Sm,anti-SSA,anti-SSB,anti-Scl-70 subgroups,and the accuracy rates were 84％,95％,98％,98％,88％,respectively.The distribution of quantitative values by different laboratories for ANA (by IIF),anti-dsDNA,anti-CCP antibody (by ELISA) varied remarkably.Conclusion Hospitals that enrolled in the survey and the items involved for quality control were increasing year by year.The accurate rates of ANA,anti-dsDNA in this survey were similar to the past national quality control surveys,the quality of AMA/ASMA,anti-CCP antibodies test was better than that of the past surveys,but the quality of anti-ENA antibodies test needs to be improved.

Objective:Generation and functional analysis of EBV LMP2A specific CTL elicited by DC transfected with recombinant adenovirus in vitro .Methods:PBMC were isolated from healthy EBV carriers and NPC patients, and then cocultured with autologous mature Ad5 LMP2A transfected DC at the ratios of 20∶1. Cytotoxicity of LMP2A specific CTL was determined with LDH release assay, the populations of CTL were performed by FACS,the IFN ? secretion and FasL mRNA expression of the CTL were detected by biological activity assays and RT PCR, respectively.Results:The results showed that high cytotoxicity of LMP2A specific CTL could be elicited by autologous transfected DC. The cytotoxicity boosted with the increase of transfected DC stimulation times, but there were no significant changes between two and three stimulations.The phenotypic analysis demonstrated that the LMP2A specific CTL induced at day 14 consisted of a majority of CD4 + and CD8 +T cells, and only a small percentage of CD56 + cells. The IFN ? secreted in the supernatants of cell culture also increased with the stimulation times. In addition, the specific CTL at day 14 from EBV healthy carriers could express FasL mRNA.Conclusion:Strong and functional EBV specific CTL could be generated by autologous mature DC transfected with adenovirus encoding LMP2A, which could provide a rationale for the immunotherapy of EBV associated NPC. [

Objective The study was to evaluate the behavioral differences in learning and memory abilities among three ani -mal models of Alzheimer′s disease. Methods Three SD rat models(n=20) were used in this study.The first SD rat model was es-tablished by intracerebral injection of Aβ25-35 into the bilateral hippocampus , the second mouse model was induced by intraperitoneal in-jection of scopolamine and the third was a senescence accelerated mouse model .Morris Water maze test was performed to investigate behavioral differences by comparison to corresponding blank control groups ( sham operation group , blank group and P8 group ) . Results The learning and memory abilities of senescence accelerated rats and scopolamine -treated rats were worse than those of the corresponding control groups, especially the scopolamine-treated rat ([35.47 ±3.78]s vs [50.61 ±3.94]s, P<0.01).There was no significant difference between Aβrats and sham-operated rats(P>0.05). Conclusion The model of Alzheimer′s disease in-duced by intraperitoneal injection of scopolamine represents more distinct changes in learning and memory abilities .Morris water maze test can be used to well evaluate whether the scopolamine-induced model is successfully established or not .

BACKGROUNDTo study the effects of dendritic cells (DC) transfected with recombinant adenovirus encoding Epstein-Barr virus(EBV)latent membrane protein 2A(LMP2A)gene, and to provide evidence for further investigation on the therapeutic vaccine against EBV-associated malignancies.

METHODSDCs were transfected with EBV-LMP2A recombinant adenovirus (Ad5-LMP2A), which was generated by homologous recombination. The expression of LMP2A protein on mature DC transfected with Ad5-LMP2A at different multiplicity of infection(MOI)was analyzed by fluorescence activated cell sorting(FACS), and the dead cells were counted by trypan blue staining. The alteration of surface markers on mature DC including CD1a, CD83, CD40, CD80, HLA?DR was detected by means of FACS before and after transfection. Meanwhile, the functions including stimulating allogenetic T cells reaction and expressing IL12 P40 mRNA on transfected DC were measured by methods of 3H-thymidine uptake and fluorescent semi?quantitative polymerase chain reaction (PCR), respectively.

RESULTSAbout 80% mature DC expressed LMP2A protein and >92% cells were viable after transfection at the MOI of 200 No. significant changes in the surface markers and the cytomorphology of mature DCs were detected during the transfection. Transfected DC still have strong potential to stimulate the proliferation of allogenetic T cells and to express IL-12 P40 mRNA.

CONCLUSIONEBV-LMP2A gene,which was carried by adenovirus vectors, could be transferred into DC with high efficiency. The function of mature DC was not affected significantly by the transfection of Ad5-LMP2A.

Adenoviridae ; genetics ; Cells, Cultured ; Dendritic Cells ; physiology ; Gene Expression ; Genetic Vectors ; Humans ; Recombination, Genetic ; Transfection ; Viral Matrix Proteins ; genetics

The aim of the present study was to investigate the involvements of insulin-like growth factor-1 (IGF-1) and estrogen receptor α (ERα) in the inhibitory effect of wogonin on the breast adenocarcinoma growth. Moreover, the effect of wogonin on the angiogenesis of chick chorioallantoic membrane (CAM) was also investigated. MCF-7 cells (human breast adenocarcinoma cell line) were subjected to several drugs, including IGF-1, wogonin and ER inhibitor ICI182780, alone or in combination. MTT assay was used to detect breast cancer proliferation. Western blot was used to analyze ERα and p-Akt expression levels. CAM models prepared from 6-day chicken eggs were employed to evaluate angiogenesis inhibition. The results showed wogonin and ICI182780 both exhibited a potent ability to blunt IGF-1-stimulated MCF-7 cell growth. Either of wogonin and ICI182780 significantly inhibited ERα and p-Akt expressions in IGF-1-treated cells. The inhibitory effect of wogonin showed no difference from that of ICI182780 on IGF-1-stimulated expressions of ERα and p-Akt. Meanwhile, wogonin at different concentrations showed significant inhibitory effect on CAM angiogenesis. These results suggest the inhibitory effect of wogonin on breast adenocarcinoma growth via inhibiting IGF-1-mediated PI3K-Akt pathway and regulating ERα expression. Furthermore, wogonin has a strong anti-angiogenic effect on CAM model.
Adenocarcinoma ; metabolism ; pathology ; Angiogenesis Inhibitors ; pharmacology ; Animals ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; Estrogen Receptor alpha ; genetics ; metabolism ; Female ; Flavanones ; pharmacology ; Humans ; Insulin-Like Growth Factor I ; antagonists & inhibitors ; pharmacology ; Scutellaria ; chemistry

OBJECTIVETo assess the effect of repeated gonadotropic stimulations on the developmental potential and growth differentiation factor-9 (GDF-9) expression of mouse oocytes.

METHODSFemale Kunming mice were treated with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (HCG) for 3 times, and the control mice were treated with normal saline. The two groups of mice were both stimulated subsequently to obtain the mature oocytes. Immunocytochemical staining was employed to evaluate GDF-9 expression in the oocytes. The oocytes were then inseminated and cultured till the formation of blastocysts to compare the cleavage rate and blastocyst formation rate between the groups.

RESULTSA total of 253 mature oocytes were obtained in the repeated stimulation group, with a mean of 11.5 oocytes from each mouse; 521 mature oocytes were obtained in the control group with a significantly greater mean number of 32.6 from each mouse (P<0.05). The average optical density and integrated optical density for GDF-9 expression were significantly lower in the oocytes in repeated stimulation group than in the control group (P<0.05 and 0.01, respectively). After insemination, the cleavage rate were comparable between repeated stimulation group and the control group (85.6% vs 88.8%), but the blastocyst formation rate was significantly lower in repeated stimulation group (20.8% vs 35.2%, P<0.01).

CONCLUSIONRepeated gonadal stimulation decreases the developmental potential of mouse oocytes possibly due to reduced GDF-9 expression.

Animals ; Cells, Cultured ; Female ; Gonadotropins ; pharmacology ; Growth Differentiation Factor 9 ; metabolism ; Mice ; Oocytes ; cytology ; growth & development ; metabolism ; Ovulation Induction ; adverse effects ; methods

Adolescent ; Adult ; Aged ; Creatine Kinase ; blood ; Female ; Hepatitis B, Chronic ; blood ; drug therapy ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Thymidine ; analogs & derivatives ; therapeutic use ; Young Adult

Objective To investigate the change of the percentage of Th17 cells and cytokine among hepatocellular carcinoma (HCC) tissues and para-carcinoma tissue and the liver tissue of hepatic hemangio-ma, and to explore its clinical significance. Methods To choose 26 of hepatocellular carcinoma patients between November 2014 and December 2016 and also include 11 cases of hepatic hemangioma as control group. The percentage of Th17cells in HCC tissue and para-carcinoma tissue and the liver tissue of hepatic hemangioma was evaluated by flow cytometric; The levels of IL-6 and IL-17 in HCC tissues, para-carcinoma tissue and the liver tissue of hepatic hemangioma were detected by RT-PCR. Results The ratio of Th17 cells to CD4 cells in peripheral blood of HCC was significantly higher than that in control group (4. 07% ± 0. 68% vs. 1. 39% ± 0. 41% ), P＜0. 05. The ratio of Th17 cells to CD4 cells in patients with hepatocellular carcinoma was significantly higher than that in paracancerous tissues and liver tissues of hepatic hemangioma (4. 76% ± 0. 75% vs. 2. 30% ± 0. 26% vs. 0. 77% ± 0. 31% , P＜0. 05). Multivariate linear stepwise regression analysis of the changes of Th17 cells in patients with HCC and their correlation with clinical parameters showed that Th17 lymphocytes in peripheral blood of patients with HCC were closely related to TNM staging(YTh17 =2. 647 +0. 687TNM. It’s indicated that the level of IL-6 and IL-17 in liver cancer tissues was significantly higher than that of the liver hemangioma in the control group through ELISA ( P＜0. 05). The proportion of peripheral blood Th17 cells to CD4 cells is different in different TNM stages(stage I＜stage II ＜stage III ～IV, the difference was statistically significant. RT-PCR analysis showed that the expression of IL-17, IL-6mRNA in hepatocellular carcinoma was significantly higher than that in adjacent tissues and hepatic hemangioma liver tissue (all P＜0. 05). Conclusion The increased proportion of Th17 cells and the increased levels of cytokines IL-6 and IL-17 may be closely correlated with occurrence and development of hepatocellular carcinoma. It can be an important target of anti-tumor therapy.

Estrogen signaling pathways play an important role in the regulation of the physiological function of breast cancer cell proliferation and apoptosis. The article used MTT assay, flow cytometer analysis and Western blot to detect the inhibition of fraxetin on MCF-7 cell cycle distribution and apoptosis, ERα, cyclin D1 and Bcl-2 expression levels, MAPK and PI3K signaling pathway to investigate the mechanism of anti-breast cancer of fraxetin. The results showed fraxetin inhibited E2β-stimulated MCF-7 cell proliferation in a dose- and time-dependent manner, reversed E2β-induced anti-apoptosis and promoted G0/G1 phase arrest. After treatment with fraxetin, the expression of ERα in MCF-7 cell was decreased, and estrogen genomic signaling pathway was inhibited by down-regulating the expression of cyclin D1 and Bcl-2 proteins. After MCF-7 cells were treated with fraxetin, the expressions of MAPK/Erk1/2 protein were reduced, which affected estrogen non-genomic signaling pathway. The results suggest fraxetin plays a part in anti-breast cancer function through E2β-mediated genomic and non-genomic signaling pathways.
Apoptosis ; Breast Neoplasms ; metabolism ; Cell Proliferation ; Coumarins ; pharmacology ; Cyclin D1 ; metabolism ; Estrogen Receptor alpha ; metabolism ; Estrogens ; pharmacology ; Humans ; MCF-7 Cells ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Signal Transduction