To investigate the different expression of melatonin receptor between adult thyroid neoplasm and its adjacent normal thyroid tissue, we extracted the total RNA of all kinds of thyroid tissue, synthesized the primer of mt 1, MT 2, and analyzed the mRNA of melatonin receptor by RT-PCR method. mt 1, MT 2 melatonin receptor subtype positive strap expressed in thyroid adenoma as well as in adenocarcinoma. MT 2 in adenocarcinoma had a higher quantity than that in normal thyroid tissue(P0.05). Therefore, mt 1 and MT 2 subtype is present in human thyroid neoplasm tissue, and MT 2 subtype is associated with the occurrence of human adenocarcinoma.

Objective To investigate the effect of MDR1 and MDR3 gene silence by shRNA of human breast carcinoma cell line MCF-7/Adr,and explore the role of MDR1 and MDR3 in adriamycin-resistance of breast carcinoma cells. Methods shRNA plasmid vector specifically targeting MDR1 and MDR3 gene was transfected into cells. The control group was transfected with empty vector. The concentration of adriamycin was detected by the flow cytometry (FCM). Cell apoptosis was analysed by FITC-Annexin-V/PI double staining. Cell viability and the IC50 of adriamycin on MCF-7/Adr cells were determined by MTT method. MDR1 and MDR3 mRNA were assessed by RT-PCR. P-gp expression was detectedby immunochemistry. Results After treatment with ABCB1 and ABCB4 shRNA plasmid vector, the apoptosis of MCF-7/Adr cells was (30.21±1.65)%and (22.07±2.17)% respectively. Compared with untransfecedgroup and empty vector transfection group the difference was significant(P＜0.01). MDR1 and MDR3 shRNAcould increase cellular adriamycin accumulation of MCF-7/Adr cells. MCF-7/Adr cells viability and the IC50were significantly decreased after transfection. Compared with untransfeced group and empty vector transfectiongroup, the mRNA level of MDR1 and MDR3 in MCF-7/Adr cells were decreased by (89.5±0.8)%and(85.1±1.2)%, the reduction of MDR1 and MDR3 mRNA was in a time-dependent manner. Immunochemistry proved that the expression of p-gp was significantly inhibited. Compared with untransfeced group and empty vector transfection group the difference was significant (P＜0.05). Conclusion The shRNA can effectively and specifically silence the expression of MDR1 and MDR3 gene, reverse the adriamycin-resistance mediated by P-gp in MCF-7/Adr cells. The reversal effect of adriamycin-resistance by shRNA of MDR1 is more effective than that of MDR3.

Objective To explore skeletal‐related events (SREs) clinical factors and analysis prognosis factors on patients with non‐small cell lung cancer(NSCLC) with bone metastases .Methods We collected clinical data of pathology confirmed 383 patients with non‐small cell lung cancer between April 2007 and January 2007 in the third affiliated hospital of the third military medical uni‐versity .It was used to screening for Emission Computed Tomography (ECT ) for bone metastases .And then it was need to con‐firmed for CT ,MRI or PET‐CT or pathology .Statistics in patients between clinical features and the SREs prediction factor with Univariate and Multivariate .And Kaplan‐Meier method analysis of survival in the non‐small cell lung cancer patients with bone me‐tastases .Results Out of 383 patients with bone metastases ,178 patients with SREs .The incidence was 46 .5% .Univariate analysis showed that women ,adenocarcinoma ,never smoking history ,single bone metastases ,bisphosphonate therapy ,targeted therapy in patients with bone metastases are less likely to have SREs ,it was considered statistically significant (P<0 .05) .Multivariate analy‐sis showed multiple bone metastases and no bisphosphonate therapy is independent risk factors for SREs .Median survival time was 14 .5 months in non‐small cell lung cancer patients with bone metastases ,1 year survival rate was 46 .5% ,2 years survival rate was 15 .9% .The survival analysis shows that more bisphosphonate treatment and bisphosphonate with EGFR‐TKI therapy on the prog‐nosis of patients with statistically significance (P<0 .05) .Conclusion It was likely to occur SREs in NSCLC patients with bone metastases .No bisphosphonate and multiple bone metastases are independent risk factors for SREs .Bisphosphonate treatment may prevent or reduce occur SREs for NSCLC patients with bone metastases ,and it may prolong survival ,it speculated that bisphospho‐nate may have resistant NSCLC cell activity .

Objective: To study the nutritional status and problems of Chinese elite athletes for instructing training scientifically. Method: 599 elite athletes were investigated by means of dietary survey, physical measure and biochemical detection. Results: (1) The daily average energy intake reached the adequate intake (AI) recommended for athletes, but the energy ratio both of protein and fat higher, carbohydrate lower, being 18.9%, 38.6% and 42.5% respectively. Except vitamin A and B1, B2 the intakes of other vitamins and minerals were sufficient. (2) The average BMI and body fat percentage were 23.0?3.0 and (12.1?3.2)% in men and 21.9?3.0 and (20.5?3.9)% in women respectively. (3) The average hemoglobin levels were (145.7?13.3)g/L in men and (130.6?11.8)g/L in women. The rates of anemia and iron deficiency anemia were 12.6% and 5.2% respectively, female higher than male. (4)The hyperlipidemia rate was 22.3%, including 12.6% high TG, 9.7% high TC and 1.2% high LDL, female higher than male. (5) The rates of VB1 and VB2 insuficiency were 46.2% and 32.7% respectively, both including 9.6% deficiency. Conclusion: The nutritional status of Chinese elite athletes was good, but still with anemia, vitamin insufficiency and hyperlipidemia.

Objective To investigate the expression of microRNA 27a (miR-27a) and relationship with drug resistance in human ovarian cancer A2780/Taxol cells.Methods A stem-loop-mediated real-time PCR was used to detect miR-27a expression in A2780 and A2780/Taxol cells.The cells were transfected with the mimics or inhibitors of miR-27a or negative control RNA ( NC) by lipofectamine 2000.The expressions of MDR1 gene,P-glycoprotein (P-gp) and homeodomain-interacting protein kinase 2 (HIPK2) protein levels were measured by real-time PCR and western blot respectively.Methyl thiazolyl tetrazolium (MTT) assay was used to analyze drug sensitivity.Apoptosis analysis was measured by fluorescence activated cell sorter ( FACS).Results (1) miR-27a was an average of 2.2-fold higher expression level in A2780/Taxol cells than that in A2780 cells,with a significant difference between the two groups (P <0.05).(2) A2780/Taxol cells transfection with inhibitors of miR-27a showed that the levels of MDR1 mRNA was decreased by 39%,P-gp protein level[(26 ±5)%]decreased than that in the NC group[(43 ±7)%],HIPK2 protein level[(30 ±6)%]increased than that in the NC group[(19 ±4)%],the 50% inhibitionconcentration (0.5 (μmol/L) was less than that in the NC group (6.8 μmol/L),apoptosis rate[(32.5 ± 3.6) %]was higher than that in the NC group[(5.6 ±2.1) %],and there were significant differences between two groups (all P < 0.05 ).( 3 ) Transfection of A2780 cells with mimics of miR-27a led to increase MDR1 mRNA expression by 121% as compared with one transfection with NC (P<0.05).Conclusion The expression of miR-27a is upregulated in A2780/Taxol cells,which may regulate MDR1 and P-gp expression by targeting HIPK2.

The expression of N-myc down-regulated gene 1 (NDRG1) has previously been reported to be involved in the proliferation, differentiation, invasion and metastasis of cancer cells, but its role in cervical cancer is still unclear. This study aimed to investigate the expression of NDRG1gene in human cervical cancer and its effect on aggressive tumor behaviors. The NDRG1 expression in cervical tissues and cells was detected by RT-PCR. Specific expression plasmid pEGFP-N1-NDRG1-GFP was used to enhance the expression of NDRG1 in human cervical cancer cell lines. The mRNA and protein level of NDRG1 was assessed by RT-PCR and Western blotting, respectively. Its effects on cell proliferation, migration, invasion, cell cycle and apoptosis were detected by MTT, transwell migration assay and flow cytometry (FCM), respectively. The results showed that the expression of NDRG1 in cervical cancer tissues and cells was significantly lower than in normal cervical tissues (P<0.001). After transfection with pEGFP-N1-NDRG1-GFP, the mRNA and protein expression of NDRG1 was up-regulated in Siha cells, which suppressed cell proliferation (P<0.001), induced cell cycle arrest (P<0.05), reduced invasion and migration of Siha cells (P<0.05), but caused no cell apoptosis. Moreover, vascular endothelial growth factor (VEGF), a tumor-induced angiogenesis factor, was markedly reduced and E-cadherin, a cell adhesion molecule, was increased in the cells transfected with pEGFP-N1-NDRG1-GFP. It was concluded that up-regulated NDRG1 may play a role in the suppression of malignant cell growth, invasion and metastasis of human cervical cancer.

Objective To compare the activities of daily living(ADL) of the elderly in different styles of providing.Methods 662 subjects were in range of 60～100 years old living in the organization for the aged and 620 subjects were in the range of 60～98 years old living at home from Beijing urban. They were evaluated with the ADL rating scales and a self-designed health status questionnaire.Results The total scores of ADL were not significantly difference between the elderly living at home and living in the organization(t=-0.299, P＞0.05). But age (OR=3.05, 95%CI: 1.805～2.935), educational level (OR=2.01, 95%CI: 1.512～2.544), and physical health (OR=1.25, 95%CI: 1.524～2.012) were related to ADL. Conclusion Age is the important factor affecting ADL (OR=3.05, 95%CI: 1.805～2.935), but ADL of the elderly in different styles of providing is not significantly difference.

Muhidrug resistance (MDR) is one of the important reasons for the failure of clinical anticancer drugs,involving multiple mechanisms.Among them,the classical MDR mechanism mediated by P-glycoprotein (P-gp) is closely related to the formation of MDR,which can excrete intracellular chemotherapeutic drugs through the "drug pump" effect and significantly reduce the therapeutic effect.Curcumin is mainly extracted from the underground rhizome of Chinese medicine turmeric,with a wide range of pharmacological activity.Recent studies have found that curcumin also has a role in reversing the MDR of the tumor,by inhibiting both P-gp function and expression,and this process involves a variety of signal paths.

To investigate the relationship between p38 mitogen-activated protein kinase (p38MAPK)and cell apoptosis during the paclitaxel resistance of ovarian carcinoma cell lines, flow cytometry(FCM) and PI staining were employed to determine the effect of p38MAPK inhibitor SB203580 onthe apoptosis of A2780/Taxol cells, a drug-resistant human ovarian carcinoma cell line. p38MAPKprotein expression in SB203580-treated cells was immunochemically measured. The 50% inhibitionconcentration (IC<,50) of paclitaxel on A2780/Taxol cells was determined by MTT assay. MDR-1mRNA, and expression of p38MAPK and phospho-p53 protein were detected by RT-PCR and West-ern blotting, respectively. The apoptosis rate of A2780/Taxol cells was (19.7±1.04)% 24 h afterSB203580 treatment. A significant difference in apoptosis rate was found among experiment group,control group and untreated group (P<0.05). The relative reversal rate of A2780/Taxol cells to pacli-taxel was (57.18±2.01)%. As compared with the control group and the untreated group, p38MAPKprotein and MDR-1 mRNA in SB203580-treated cells was substantially decreased. The expression ofp53 protein was significantly increased. It is concluded that p38MAPK pathway is related to pacli-taxel resistance of ovarian carcinoma, and blockade of this pathway can promote the apoptosis of thedrug-resistant cells and reverse the drug-resistance. Moreover, p38MAPK-mediated apoptosis in pa-clitaxel-resistant ovarian carcinoma cells depends on the activation of p53.

OBJECTIVETo establish red-green dual-color fluorescence glioma model in nude mice and to explore its practical values.

METHODSCM-DiI-stained rat glioma C6 cells (C6-CM- DiI cells) expressing red fluorescence were inoculated into the brain of athymic nude mice expressing green fluorescence protein (NC-C57BL/6J-EGFP). Then the whole-body dual-color fluorescence imaging was detected dynamically. Finally whole brains of the tumor-bearing mice were removed and 5 µm thick serial frozen slices were made. Light microscopy, fluorescence microscopy and confocal laser scanning microscopy were performed to observe the transplanted tumor tissue structure and fluorescent cells.

RESULTSTumor mass with red fluorescence increased gradually under continuous in-vivo fluorescence imaging monitoring. Under the fluorescence microscope, cells with red, green and yellow fluorescence were observed in the frozen sections of transplanted tumor tissue and the mutual structural relationship among them could be defined. The tumor cells migration, implantation and cell fusion between transplanted tumor cells and host cells could be observed. It could be distinguished according to the fluorescence, that blood vessels of tumor-origin displayed red fluorescence, blood vessels of host-origin displayed green fluorescence and mosaic blood vessels appeared yellow fluorescence. It was depicted that host innate astrocytes and oligodendrocytes in the microenvironment at the tumor periphery could be activated and dedifferentiated into nestin-positive cells.

CONCLUSIONSIn contrast to traditional animal model, the dual-color fluorescence imaging of nude mouse models of glioma possesses enormous advantages in investigating tumor mass in-vivo fluorescence imaging, tumor cells migration and metastasis, tumor angiogenesis and reactive activation of host innate cells in the microenvironment at tumor periphery, thus, has highly practical application value.

Animals ; Astrocytes ; metabolism ; Brain Neoplasms ; blood supply ; metabolism ; pathology ; ultrastructure ; Carbocyanines ; metabolism ; Cell Fusion ; Cell Line, Tumor ; Cell Movement ; Disease Models, Animal ; Fluorescent Dyes ; metabolism ; Glioma ; blood supply ; metabolism ; pathology ; ultrastructure ; Green Fluorescent Proteins ; metabolism ; Luminescent Proteins ; metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Nude ; Microscopy, Confocal ; Microscopy, Fluorescence ; Neoplasm Transplantation ; Neovascularization, Pathologic ; Nestin ; metabolism ; Oligodendroglia ; metabolism ; Rats