Objective RNPC1 may act as an oncogene or suppressor gene in human tumors and its role in human renal cell carcinoma (RCC) remains unclear.The objective of this study was to investigate the role of RNPC1 in the development of RCC.Methods Over-expression of RNPC1 gene group (RNPC1 group) and short hairpin RNA interfering RNPC1 gene expression (shRNPC1 group) were respectively built in RCC CAKI-1 and CAKI-2.The blank control group (NC group) and negative control group (SCR group) were built as well.The qRT-PCR and western blot (WB) were used to detect the expression levels of RNPC1 mRNA and RNPC1 protein in RCC cells.Lentivirus infection was applied to establish stable expressed RCC cell lines of RNPC1 over-expression and interference.Detection was made on mRNA and protein expression levels in RNPC1 stable RCC cell lines.The effects of RNPC1 on cell proliferation, colony formation assay, migration, and invasion were detected by CCK-8 cell differentiation test, clone test, scratch test, and migration and invasion test.WB was applied to detect the change of protein expression in the EMT path of RNPC1 stable RCC cell lines and explore the molecular mechanism of RNPC1 effect on the biological function of RCC cells.Results The expression levels of RNPC1 mRNA and protein were found lower in shRNPC1 group than those in SCR group, while the expression levels of RNPC1 mRNA and protein in SCR group were higher than those NC group (P<0.05).The capability of proliferation in shRNPC1 group was stronger than that in SCR group, while the capability of proliferation in shRNPC1 group was weaker than that in NC group (P<0.05).The capabilities of cell migration and invasion were stronger in shRNPC1 group than those in SCR group, while the capabilities of cell migration and invasion in RNPC1 group were weaker than those in NC group (P<0.05).RNPC1 could inhibit the proliferation capability of RCC cells and might up-regulate the protein expression of E-cadherin and down-regulate the protein expression of β-catenin and vimentin, thus inhibiting EMT path and the capabilities of migration and invasion off RCC cells (P<0.05).Conclusion RNPC1 acts as a tumor suppressor in RCC and has the potential for the prediction of RCC prognosis.

AIM: To study effects of urokinase-type plasminogen activator (uPA) signal transduction on expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-3 (TIMP-3) in giant cell tumor of bone (GCT). METHODS: Expression of uPAR, MMP-2 and TIMP-3 in GCT tissue was detected by immunohistochemistry. Phosphorylation level of mitogen-activated protein kinase (p44) in uPA/uPAR signal pathway in cultured GCT cells was detected by immunoprecipitation. The expression of MMP-2 and TIMP-3 in cultured cells after treatment with uPA-ATF or anti-uPAR antibody was also detected by Western blotting. RESULTS: 1) Urokinase-type plasminogen activator receptor (uPAR) was positive on the cell membrane and in cytoplasm of some mononuclear stromal cells (MSCs) and multinucleated giant cells (MGCs); 2) MMP-2 was positive in the cytoplasm and on the cell membrane of almost all of MSCs and some of MGCs. The polar distribution of MMP-2 in the cytoplasm of MGCs was especially obvious; 3) The expression of TIMP-3 of some MSCs and MGCs in GCT was much lower than MMP-2. The positive signal also showed a prominent polarity; 4) After treatment with uPA-ATF, the phosphorylation level of p44 in GCT cultured cells was much higher than the control. Addition of anti-uPAR antibody in the cells remarkably down-regulated the phosphorylation level of p44 as compared with the control group, suggesting that uPA-ATF participates cell signal transduction and this reaction can be inhibited by anti-uPAR antibody; 5) uPA-ATF cell signal pathway up-regulated expression of MMP-2 and TIMP-3, while anti-uPAR antibody down-regulated the expression of MMP-2 and TIMP-3. CONCLUSION: These results demonstrate for the first time that uPA-ATF directly regulates the expression of MMP-2 and TIMP-3 by signal transduction pathway, and the over-expression of MMP-2 and TIMP-3 may play an important role in local osteolysis of GCT. [

Objective To observe the effect of high iodine on mRNA expression of thyroid hormone receptor (TSHR),protein kinase A (PKA) and sodium iodide symporter (NIS) in peripheral blood of patients with thyroid diseases during lacatation.Methods A total of 99 breast-feeding women were selected as observation objects in Shanxi Province's sufficient iodine and high iodine areas,and they were divided into case group and control group according to whether suffer from thyroid disease.In high iodine areas,there were 21 patients and 19 healthy controls.In sufficient iodine areas,there were 30 patients and 29 healthy controls.Peripheral blood of all the observation objects was collected,and mRNA expression of TSHR,PKA and NIS was detected by real-time quantitative PCR.Results The case group [median (M):0.099,0.994] and the control group (M:0.240,0.738) in the high iodine areas were respectively compared with the case group (M:3.087,1.127) and the control group (M:1.823,0.842) in the sufficient iodine areas.The TSHR mRNA expression was significantly decreased (Z =-5.034,-4.010,all P ＜ 0.01);the PKA mRNA expression had a downward trend,and the difference was not statistically significant (Z =2.895,-0.343,all P＞ 0.05).The NIS mRNA expression of the case group in high iodine areas (M:0.485) was obviously lower than that of the the case group in sufficient iodine regions (M:2.680,Z=-3.311,P ＜ 0.01).The control group in high iodine areas (M:0.470) was compared with the control group in sufficient iodine areas (M:0.835),and the difference was not statistically significant (Z =-1.882,P ＞ 0.05).The NIS and the TSHR mRNA were positively correlated [correlation coefficient (r) =0.741,P ＜ 0.01];the NIS and the PKA mRNA was also positively correlated (r =0.293,P ＜ 0.01);but the TSHR mRNA was not significantly correlated with the PKA mRNA (r =-0.081,P ＞ 0.05).Conclusion Lactating women may have protected themselves and their babies through TSH-TSHR-cAMP-PKA signaling pathway that regulating iodine levels.

Objective To investigate the change of serum Tg levels of DTC patients with positive stimulated Tg (Tg ≥ 10.00 μg/L),negative 131I-diagnostic whole body scan(Dx-WBS) and no distant metastasis 6 months after initial 131I therapy.Methods Fifty-six DTC patients (20 males,36 females,average age 43.11 (21-70) y) with intermediate or low risk of recurrence according to American Thyroid Association (ATA) guideline were enrolled into the retrospective study.All patients were grouped according to stimulated Tg level after initial 131I therapy:group with positive Tg (Tg+ group,n =19) and group with negative Tg (Tgˉ group,n=37).Changes of suppressed Tg at 1 year and 2.5 years (Tg1ysup and Tg2.5ysup) after initial therapy were compared between the two groups.Serum TSH level,TgAb level,neck ultrasound and chest CT results were also evaluated.The two-sample t test and x2 test were used for statistical analysis with SPSS 17.0.Results Stimulated Tg and Tglysup levels in Tg+ group were remarkably higher than those in Tgˉ group:(24.27±4.10) μg/L vs (2.73±3.01) μg/L,t=7.191,P＜0.05(6 months after initial 131I therapy) ; (2.21±0.55) vs (0.48±0.10) μg/L,t=3.102,P＜0.05(1 year after initial 131I therapy),respectively.In Tg+ group,suppressed Tg level decreased with time in 68.4％ (13/19) of patients,of whom the Tg2.5ysup level was much lower than Tglysup level ((0.53±0.15) μg/L vs (1.38±0.50) μg/L).Tg2.5sup level in Tg+ group became comparable to that in Tgˉ group ((1.44±0.52) μg/L vs (0.38±0.07) μg/L; t =2.001,P＞0.05).In each group,one case of recurrence with suppressed Tg of 1.4 μg/L and 0.1 μg/L respectively,was observed using neck ultrasound after 2 years of follow-up.Conclusions Serum Tg levels decreased with time for Tg+/131I-Dx-WBS-DTC patients with intermediate or low risk of recurrence.It might not be necessary to follow up these patients with Tg and 131 I-DxWBS after 6 months of initial 131I therapy.

The focus of microbiologists has moved to the rare actinomycetes.For selective isolation of rare actinomycetes that all play the important role in bioactive compounds,the approaches which involve the methods using gellan gum and flooding solution、 rehydration-centrifugation(RC)、 extremely high frequency radiation(EHF)、 bacteriophage and sucrose-gradient centrifugation were introduced in this paper.

OBJECTIVETo analyze alterations in the gene expression profiles of Velcade-treated K562 cells using bioinformatics methods.

METHODSThe total RNAs of Velcade-treated and control K562 cells were amplified and labeled with fluorescent dyes. The labeled RNAs were hybridized to Agilent Human 1A Microarray, and the raw expression data were processed with Agilent Feature Extraction Software. GeneSifter and GATHER were used for data analysis of the differentially expressed genes to perform gene ontology classification, KEGG pathway analysis, functional protein association network construction and literature mining.

RESULTSTotally 228 differentially expressed genes were identified in the Velcade-treated K562 cells. including 84 up-regulated and 144 down-regulated genes. Chymase 1 gene had the greatest down-regulation by 10.80 folds (log ratio), and interferon alpha-21 gene was also down-regulated by 2.31 folds. Gene ontology classification suggested enhanced aging and leukocyte activity. KEGG pathway analysis showed significant impact of Velcade on JAK-STAT signaling pathway, cytotoxicity mediated by natural killer cells, and antigen processing and presentation pathways. Protein-protein interaction analysis revealed that ubiquitin-dependent protein catabolism, antigen presentation and immune response, as well as JAK-STAT signaling pathway were the major elements of the protein network. Literature mining showed that the differentially expressed genes were strongly associated with terms such as leukemia, apoptosis, cell cycle, proteasome, inhibitor, aging and IkappaB, etc.

CONCLUSIONSVelcade may inhibit the cell survival pathways such as NF-kappaB and JAK-STAT signaling pathways to enhance the cytotoxicity and inducing tumor cell apoptosis. Velcade might also be involved in antigen processing and presentation, immune response and inflammation. Chymase 1 gene is probably the key target of Velcade.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Boronic Acids ; pharmacology ; Bortezomib ; Chymases ; genetics ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; K562 Cells ; Oligonucleotide Array Sequence Analysis ; methods ; Pyrazines ; pharmacology

OBJECTIVETo observe the effect of electric acupuncture (EA) on the Nogo receptors (NgR) protein expression in the cerebral cortex, the medulla oblongata, and the spinal cord of cerebral ischemia-reperfusion (I/R) stroke-prone renovascular hypertensive rats (RHRSP) with middle cerebral artery occlusion (MCAO) at different time points, and to investigate its possible mechanisms for remote-organ injury of acute cerebral infarction (ACI).

METHODSThe RHRSP model was duplicated in male SPF grade SD rats. Then the MCAO model was prepared by a thread stringing method. Rats were divided into the hypertension group,the sham-operation group, the MCAO group, the EA group, and the sham-acupoint group by random number table method, 60 in each group. Rats in the MCAO group only received MCAO reperfusion treatment. Those in the sham-operation group only received surgical trauma. Baihui (DU20) and Dazhui (DU14) were needled in the EA group, once daily for a total of 28 days.The needles were acupunctured at the skin one cun distant from Baihui (DU20) and Dazhui (DU14) and then the same EA treatment was performed in the sham-acupoint group. At day 1, 7, 14, 28 after treatment, six rats were executed from each group, and their right cortex and medulla oblongata, and the left spinal cord were isolated. The infarct volume was detected by Nissl's staining method. The NgR expression was detect by Western blot.

RESULTS(1) In the cortex area: compared with the hypertension group,the NgR expression increased in the MCAO group at day 1,7,14,and 28 after MCAO (P < 0.05). Compared with the MCAO group, the NgR expression of the EA group and the sham-acupoint group were equivalent at 1 day af ter MCAO (P > 0.05). At day 7, 14,and 28 after MCAO, the NgR expression decreased in the EA group (P < 0.05), it was quite similar to that in the sham-acupoint group (P > 0.05). (2) In the medulla oblongata area: compared with the hypertension group, the NgR expression was equivalent in the sham-operation group. the MCAO group,the EA group, and the sham-acupoint group at 1 day after MCAO (P > 0.05). At day 7.14, and 28 after MCAO, the NgR expression increased in the MCAO group (P < 0.05). Compared with the MCAO group,the NgR expression decreased in the EA group at day 7, 14, and 28 after MCAO (P < 0.05), whereas it was similar in the sham-acupoint group (P > 0.05). (3) In the spinal cord area: compared with the hypertension group, the NgR expression was equivalent in the sham-operation group, the MCAO group,the EA group, and the sham-acupoint group at day 1 and 7 after MCAO (P > 0.05). At day 14 and 28 after MCAO, the NgR expression increased in the MCAO group (P < 0.05). Compared with the MCAO group, the NgR expression decreased in the EA group at day 14 and 28 after MCAO (P < 0.05), whereas it was equivalent in the sham-acupoint group (P > 0.05).

CONCLUSIONSIncreased NgR expression in the cerebral cortex, the medulla oblongata, and the spinal cord of cerebral infarct rats was an important reason for involving remote-organ injury of ACI. The protective effect of EA on hypertensive I/R cerebral injury rats might be closely related to down-regulating central nervous system myelin growth inhibition mediated factors Nogo-A receptor NgR protein expression.

Animals ; Cerebral Infarction ; metabolism ; therapy ; Disease Models, Animal ; Electroacupuncture ; GPI-Linked Proteins ; metabolism ; Hypertension, Renal ; metabolism ; therapy ; Male ; Medulla Oblongata ; metabolism ; Myelin Proteins ; metabolism ; Nogo Receptor 1 ; Rats ; Rats, Sprague-Dawley ; Receptors, Cell Surface ; metabolism ; Spinal Cord ; metabolism

Emerging data indicated that HCV subverts the antiviral activity of interferon (IF); however,whether HCV core protein contributes to the process remains controversial. In the present study, we examined the effect of HCV core protein on interferon-induced antiviral gene expression and whether the effect is involved in the activation and negative regulation of the Jak/STAT signaling pathway. Our results showed that, following treatment with IFN-α, the transcription of PKR, MxA and 2'-5'OAS were down-regulated in HepG2 cells expressing the core protein. In the presence of HCV core protein,ISRE-dependent luciferase activity also decreased. Further study indicated that the core protein could inhibit the tyrosine phosphorylation of STAT1, whereas the level of STAT1 expression was unchanged.Accordingly, SOCS3, the negative regulator of the Jak/STAT pathway, was induced by HCV core protein. These results suggests that HCV core protein may interfere with the expression of some interferon-induced antiviral genes by inhibiting STAT1 phosphorylation and induction of SOCS3.

Background Cellular autophagy is a non-apoptosis death form of tumor tissue.Research determined that arsenie trioxide (As2O3) leads to apoptosis of tumor cells.But whether As2O3 induce autophagy of SO-Rb50 cells or not is unclear.Objective This study was to assess the effects of As2O3 on autophagy of SO-Rb50 cells.Methods As2O3 with the concentration of 0,0.5,1.0,2.0,4.0 μmol/L was used to treat the SO-Rb50 cell line for 48 hours,and the growth and proliferation of SO-Rb50 cells were detected using MTT assay (A570).pGFP-LC3,a marker of autophagy,was constructed to transfer SO-Rb50 cells,and the cells were then divided into RPMI-1640 culture group (untreated group),As2O3 + RPMI-1640 culture group (As2O3 treated group) and rapamycin culture group (positive control group).Autophagy of SO-Rb50 cells was examined by laser confocal microscope and monodansylcadaverine (MDC) influorescence staining,respectively,48 hours following cell culture.Ultrastructural features of autophagy were examined with transmission electron microscope (TEM).The percentage of autophagy positive cells in different concentrations of As2O3 treated groups was calculated with flow cytometer.Results The A570 values of SO-Rb50 cells were 2.194±0.066,1.841 ±0.213,1.035±0.046,0.374±0.042 and 0.167±0.019 in 0,0.5,1.0,2.0,4.0 μmol/L As2O3 treated groups,with a significant difference among these 5 groups(F=547.636,P＜0.05),and those of 0.5,1.0,2.0,4.0 μmol/L As2O3 treated groups were significantly reduced in comparison with untreated group (P =0.000).The positive granular spots for GFP-LC3 chimeric protein were seen to aggregate in autophagic vacuoles in the As2O3 treated group and positive control group,but diffuse cytoplasmic signal for GFP-LC3 was found in the untreated group.Normal ultrastructure of SO-Rb50 cells was exhibited in the untreated group,and many double-membrane-like bound vesicles and autlysosomes were documented in the As2O3 treated group and positive control group under the TEM.A lots of MDC fluorescence granule were found in the As2O3 treated group and positive control group rather than the untreated group.Flow cytometry showed that the percentages of SO-Rb50 cells were 0,15.6％,42.7％,57.9％,79.5％ and 89.0％ in the 0,0.5,1.0,2.0,4.0 μmol/L As2O3 groups and positive control group,respectively,showing a As2O3 concentration-dependent increase.Conclusions As2O3 can induce the autophagy of SO-Rb50 cells and inhibit the proliferation of SO-Rb50 cells.Autophagic response of SO-Rb50 cells appears prior to the nuclear change after exposed to As2O3.The degree of autophagy of SO-Rb50 cells is associated with As2O3 dose.

BACKGROUNDBRAF(V600E) mutation is correlated with local aggressive clinicopathological features in papillary thyroid carcinoma; yet the relationship between this genetic variation and distant papillary thyroid carcinoma metastasis was unclear. This study aimed to investigate whether BRAF(V600E) is predictive for distant metastasis in the Chinese population.

METHODSOne hundred and seven patients with papillary thyroid carcinoma were enrolled in this study, including 43 patients with distant metastasis and 64 patients without. Quantitative real-time polymerase chain reaction was used to detect BRAF(V600E) mutation, while immunohistochemistry was performed to detect vascular endothelial growth factor (VEGF) expression. The associations between distant metastasis and BRAF(V600E) mutation, and VEGF expression as well as local clinicopathological factors were determined.

RESULTSA total of 28.6% of the patients in the distant metastasis group harbored BRAF(V600E) mutation, which was significantly lower than in the without distant metastasis group (68.8%, P < 0.001). BRAF(V600E) mutation was negatively correlated with positive VEGF expression (P = 0.001). Furthermore, 52.2% of the patients with distant metastasis exhibited VEGF expression, compared with 25.0% of those without. Higher levels of VEGF expression were also observed in the distant metastasis group. Tumor size, extra-thyroid invasion, and BRAF(V600E) mutation were independent predictors for distant metastasis according to multivariate analysis (odds ratios were 2.8, 12.4, and 0.3; 95% CI 1.483-5.334, and 2.950-52.407, 0.100-0.890; P = 0.002, 0.001, and 0.030, respectively). BRAF(V600E) mutation was negatively correlated with distant metastasis in adult subgroup analysis (P = 0.005) but was not an independent parameter.

CONCLUSIONSBRAF(V600E) mutation is predictive for distant metastasis in papillary thyroid carcinoma but not positively. VEGF may be involved in the pathogenesis of distant metastasis.

Adult ; Carcinoma ; chemistry ; genetics ; pathology ; Carcinoma, Papillary ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Neoplasm Metastasis ; Proto-Oncogene Proteins B-raf ; genetics ; Thyroid Neoplasms ; chemistry ; genetics ; pathology ; Vascular Endothelial Growth Factor A ; analysis