Objective To investigate effect on telomerase activity, apoptosis and p53/bcl-2 gene expression of MCF-7 cells line induced by paclitaxel.Methods By techniques of cell culture in vitro, telomeric repeat amplification protocol with ELISA(TRAP-ELISA) and flow-cytometry (FCM), MCF-7 cell line was treated by paclitaxel in various concentration for 72h.Results Paclitaxel down-regulated telomerase activity of MCF-7 cell, induced apoptosis of the cell in a concentration-dependent manner, significantly decreased expression of bcl-2 gene protein and increased expression of p53 gene protein. There was a positive correlation between telomerase activity and apoptosis and the expression of p53/bcl-2 gene protein.ConclusionPaclitaxel could down-regulate telomerase activity,induce apoptosis, decrease expression of bcl-2 gene protein and increase expression of p53 gene protein, which may be one of important mechanisms of Paclitaxel′s anticancer action.

The culture of umbilical cord-derived mesenchymal stem cells is extremely important for studies on umbilical cord mesenchymal stem cells. Optimization of cellculture technology is crucial for clinical application of mesenchymal stem cells and even celltherapy. Meanwhile, the labeling and tracer technique of umbilical cord-derived mesenchymal stem cells is a hotspot in stem celltransplantation. OBJECTIVE:To review the research and development of the cellmarkers and tracer methods of umbilical cord-derived mesenchymal stem cells. METHODS:A computer-based search of VIP, CNKI, Medline, Highwire and Foreign Journals Integration System databases was performed for articles concerning culture and labeling of umbilical cord-derived mesenchymal stem cells published from January 2001 to October 2013. The keywords were“stem cells, mesenchymal stem cells, umbilical cord-derived mesenchymalstem cells, cellculture, labeling methods”in Chinese and English, respectively. Final y, 35 articles were included in result analysis. RESULTS AND CONCLUSION:Umbilical cord-derived mesenchymal stem cells have not yet been widely used, mainly because of the immature isolation, culture and staining techniques of umbilical cord-derived mesenchymal stem cells. These techniques are worthy of further optimization studies. Although in recent years, cellmarkers and tracer technology of umbilical cord-derived mesenchymal stem cell s have made great progress, there are stil many problems need to be solved.

Objective To evaluate the clinical value of 99Tcm-MIBI scintigraphy for diagnosis of residual thyroid tissue and metastasis in patients with DTC after their first 131I therapy.Methods From February 2010 to March 2014,192 DTC patients (38 males,154 females,average age (43.2±8.6) years) who received total or near-total thyroidectomy and were pathologically diagnosed as DTC (171 papillary and 21 follicular carcinomas) underwent 99Tcm-MIBI scan (average dosage:740-925 MBq) 6 months after their first 131 I therapy.131 I scans was performed 4 d after oral administration of 131I of therapeutic dose (average dosage:5 550-8 140 MBq).Pre-and post-therapeutic images and the serum Tg level (detected before the imagings) were compared and analyzed.Any abnormal uptake of agent found inside or outside the thyroid was regarded as positive result.Patient-based and lesion-based data analysis were performed by x2 test and two-sample t test.Results A total of 191 patients were finally included,of which 65 positive cases were found.The sensitivity of 99Tcm-MIBI imaging was significantly lower than that of 131I imaging(56.9％ (37/ 65) vs 92.3％ (60/65);x2 =14.7,P＜0.01).Among 43 thyroid remnants and 22 metastatic lesions,99Tcm-MIBI imaging detected 39.5％ (17/43) of thyroid remnants and 90.9％ (20/22) of metastases,and those of 131I imaging were 100％ (43/43) and 77.3％ (17/22) respectively.The sensitivity of 131 I imaging in detecting thyroid remnants was significantly higher than that of 99Tcm-MIBI imaging(x2=24.0,P＜0.01).The sensitivities in detecting metastasis were not significantly different (x2=0.57,P＞0.05).The serum Tg level of positive groups (99Tcm-MIBI positive + 131I positive or 131I negative) were significantly higher than that of 99Tcm-MIBI negative + 131I negative group (t =-20.7 and-6.0,both P＜0.01),and that of 99Tcm-MIBI positive + 131I negative group was higher than that of 131I positive + 99Tcm-MIBI negative group(t=-2.7,P＜0.05).Conclusion 99Tcm-MIBI imaging could detect metastasis of DTC patient after first radioiodine therapy,but the value in detecting thyroid remnants is limited.

BACKGROUND: Antiviral drugs can reduce the incidence of early-onset cytomegalovirus(CMV)disease,but are associated with strong toxicity and the development of late-onset CMV disease.In order to prevent CMV disease better,cytotoxic T lymphocytes(CTL)may play a critical role in controlling CMV reactivation.Fluorescent HLA-peptide tetramers are used to monitor the recovery of CMV CTL in recipients of allogeneic transplants.OBJECTIVE: To explore the effect of HLA-peptide tetramers and adoptive immunotherapy in treating CMV disease.METHODS: A computer-based online search of Pubmed and Wanfang databases was performed for articles related to CTL detection,application of antiviral drugs and HLA-peptide tetramers,and adoptive immunotherapy with key words"HLA-peptide tetramers,cytomegalovirus,specific CTL,adoptive immunotherapy"in English and Chinese.Repetitive articles were excluded and 29 articles were included.RESULTS AND CONCLUSION: Adoptive immunotherapy with CMVs cytotoxic T cells as preemptive therapy is a very elegant strategy; however,generation of these cells is costly and time-consuming,and therefore the therapy is not available at every transplantation center.Magnetic selection of CMVs CD8+T cells from peripheral blood of CMV-seropositive donors by using HLA-peptide tetramers may be very hopeful,which simplifies adoptive immunotherapy.

Objective: To reconstruct adenovirus vector of breast eukaryotic initiation factor 4E and to observe its effect on the metastasis ability of breast cancer cell line MCF-7. Methods: eIF-4E gene was constructed into adenovirus vector pAD-X by gene recombination technique, which was transformed into 293 packaged cell for high titer adenovirus. Real-time PCR was applied to detect eIF-4E gene expression. eIF-4E siRNA was applied and then transwell cabin assay was used to observe changes of invasion and motor ability of MCF-7 cells transfected with reconstruction adenovirus. Result:The finding of digestion was coincided with expected. eIF-4E gene over-expression was detected in transfected MCF-7 cells with real-time PCR. And the invasion and motor abilities of transfected MCF-7 cells were more significantly inhibited in transwell cabin assay (respectively p

Objective To study the alteration of telomerase activity of MCF 7 cell line of breast cancer in the presence of different anticancer drugs. Methods The hyperplasia and viability of MCF 7 cell were detected by cell counting and trypan blue exclusion, and the telomerase activity was measured by TRAP.The alteration of MCF 7 cell and its related factors of telomerase activity were observed on cell growing in different condition. Results In abscence of drug, there was a positive correlation between hyperplasia and telomerase activity of the cell(r= 0.901 ). Adriamycin, paclitaxel and cisplatin could obviously inhibit the growth and reduce the telomerase activity of the cell in a dose-dependent and time-dependent fashion, and this reduction in telomerase activity closely correlated with the reduction in the number of viable cells. Conclusions Adriamycin, paclitaxel and cisplatin can inhibit the growth of MCF 7 cell, which may be correlated with the reduction in telomerase activity and cell viability.

This paper discussed that how to cultivate high-quality medical talents by the use of basic education,medical moral education and psychologic education etc.

Objective To establish a method for isolation of cynomolgus monkey umbilical cord mesenchymal stem cells.Methods Fresh cynomolgus monkey umbilical cord was directly minced into pasty fine pieces, and the pieces were cultured in tissue flask with DMEM/F12 medium supplemented with 10% fetal bovine serum.The morphological characteristics of the resulting cells were examined, and their expression of mesenchymal cell surface markers were analyzed by flow cytometry.The multidifferentiation potential was examined in vitro, too.Results The fibroblast-like cells were successfully isolated from the fresh umbilical cord by an adherent culture procedure.These adherent cells expressed mesenchymal markers including CD29, CD44, and CD90, and also could be induced to differentiate into adipocytes, osteoblasts and chondrocytes.Conclusion Mesenchymal stem cells can be isolated from fresh cynomolgus monkey umbilical cord by using an adherent culture procedure.

Objective To investigate the changes of circulating miRNAs expression profiles in different subtypes of ischemic stroke according to the Trial of Org 10 172 in Acute Stroke Treatment.Methods We selected 16 patients diagnosed as acute ischemic stroke at first time in the Department of Neurology,the Affiliated Hospital of Medical College Qingdao University from November to December in 2012.They were divided into large artery atherosclerosis (LAA) stroke group (n =8) and small artery occlusive (SAO) stroke group (n =8).At the same time,8 healthy checkup subjects were selected as control.High-throughput sequencing was used to detect the expression profiles of miRNAs in the plasma,and the high-throughput sequencing results were validated by quantitative real-time polymerase chain reaction.We performed the miRNAs variance analysis and associated bioinformatics analysis.Results Thirty hundred and sixty-nine miRNAs were detected in LAA group,SAO group and the control group.We found remarkable differences (fold change ＞2,P ＜0.01) in the expression of 12 miRNAs,including let-7a-5p,miR-744-5p,etc.,between any two groups of the three groups.Thirty-four miRNAs,containing miR-126-5p,miR-23a-3p,miR-143-3p,etc.,had a lower expression (fold change ＞2,P ＜0.01)in LAA group than that in the control group.In comparison with the control group,miR-1304-3p and miR-451a were significantly down-regulated (fold change ＞ 2,P ＜ O.01),while 27 miRNAs were significantly up-regulated (fold change ＞2,P ＜0.01) in SAO group.The expression levels of miR-146b-5p,miR-23a-3p and miR-451 a were validated in accordance with the results of real-time PCR.Target gene prediction and functional analysis revealed that target genes regulated by differential expression of miRNAs were mainly associated with cell proliferation,adhesion,metabolism and other biological functions.Conclusion miRNAs are differently expressed in the plasma of LAA group,SAO group and healthy control group,which suggest that miRNAs might play different roles in the pathogenesis of LAA stroke and SAO stroke by regulating downstream target genes.

BACKGROUND:There are less studies addressing whether bone marrow mesenchymal stem cells from systemic lupus erythematosus patients are different from healthy people.
OBJECTIVE:To compare the multi-differentiation capacity of bone marrow mesenchymal stem cells isolated from systemic lupus erythematosus model mice and normal control mice.
METHODS:Bone marrow mesenchymal stem cells of systemic lupus erythematosus model mice and C57BL mice were isolated and cultured fol owed by osteogenic and adipogenic differentiations, respectively. Differentiation abilities of two kinds of mouse bone marrow mesenchymal stem cells were observed.
RESULTS AND CONCLUSION:Passaged bone marrow mesenchymal stem cells from C57BL mice were long spindle-shaped and evenly distributed, while bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice showed slow growth and were relatively smal er than those from C57BL mice. After osteogenic induction, the amount of calcium salt and calcium nodules were significantly less in the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice than from normal control mice. PCR detection showed that expressions of Runx2, alkaline phosphatase and osteocalcin were also significantly decreased in the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice. After adipogenic induction, the number of lipid droplets in the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice was significantly less than the control group, and PCR detection also showed significantly decreased expression of adipogenic genes, including PPARγ2 and lipoprotein lipase. These findings suggest that the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice exhibit lower osteogenic and adipogenic capacities than those from normal C57BL mice, and also have the impaired multi-differentiation ability.