[Effect of Cyr61 on Imatinib Resistance in Chronic Myeloid Leukemia and Its Mechanism].

OBJECTIVE: To investigate the effect of Cyr61 on imatinib (IM) resistance in chronic myeloid leukemia (CML) and its mechanism. METHODS: Cyr61 level in cell culture supernatant was determined by enzyme-linked immunosorbent assay. The expression of Cyr61 and Bcl-xL were measured by real-time PCR and Western blot. Cell apoptosis was analyzed using an Annexin V-APC Kit. Expression of signal pathways related proteins was determined by Western blot. RESULTS: The level of Cyr61 obviously increased in K562G cells (IM resistance to CML cell line K562). Down-regulating the expression of Cyr61 decreased the resistance of K562G cells to IM and promoted IM induced apoptosis. In CML mouse model, down-regulating the expression of Cyr61 could increase the sensitivity of K562G cells to IM. The mechanism studies showed that Cyr61 mediated IM resistance in CML cells was related to the regulation of ERK1/2 pathways and apoptosis related molecule Bcl-xL by Cyr61. CONCLUSION: Cyr61 plays an important role in promoting IM resistance of CML cells. Targeting Cyr61 or its related effectors pathways may be one of the ways to overcome IM resistance of CML cells.

[Myxoid liposarcoma of the testis: A case report and review of the literature].

OBJECTIVE: To investigate the clinicopathological features, diagnosis, differential diagnosis, treatment and prognosis of myxoid liposarcoma (MLPS)of the testis. METHODS: We observed the clinicopathological and immunophenotypic characteristics of primary MLPS of the testis in a male patient and reviewed the relevant literature. RESULTS: The tumor was located in the right testicular parenchyma, of multi-nodular growth microscopically. The tumor cells were stellate, fusiform, foamy and signet ring-like, with abundant mucus and a network of fine branched or plexiform capillaries in the stroma. Immune phenotype-related findings included positive p16 and H3K27ME3 tumor cells and CD34 vascular network, negative CD99, CK, Desmin, FLI agreed-1, S-100, SMA, STAT6, TFE3, α-inhibin and SOX1, and a Ki-67 proliferation index of about 15%. CONCLUSION: Primary MLPS of the testis is rare, which needs to be differentiated from myxosarcoma, rhabdomyosarcoma, myxoma, spindle cell or pleomorphic liposarcoma, and lipophiloma. Immunohistochemical markers Vimentin, s-100 and CD34 can assist in the diagnosis of the tumor.

Transcriptional regulation of the porcine miR-17-92 cluster.

The miR-17-92 cluster has been involved in the cell cycle, apoptosis, and signaling. However, its transcriptional regulation has not been fully characterized. To elucidate the transcriptional regulation, the promoter of miR-17-92 was analyzed in detail in pig here. We found that, as an intronic miRNA, porcine miR-17-92 cluster was regulated by two independent promoters, an A/T-rich region directly upstream of the miR-17-92 coding sequence, and a G/C-rich region corresponding to the host gene promoter of the human miR-17-92 cluster. Several cis-regulatory elements were identified including sites for c-Myc, NFY, E2F3, and SP1, among which NFY and c-Myc sites were present in both A/T- and G/C-rich regions, while E2F3 and SP1 sites only existed in G/C-rich region. Sites for c-Myc, E2F3, and SP1 were positive for regulating transcription. NFY sites played bipartite roles, functioning as a repressor for the A/T-rich region, and as an activator for the G/C-rich region. Additionally, we found that levels of individual miRNAs in the cluster were not promoted completely in parallel with each other or with pri-miR-17-92 by the A/T-rich region, through using a self-made vector by modifying pGL3-basic in which firefly luciferase gene was replaced with an miR-17-92 cluster and a direct upstream A/T-rich region. The expression regulation of miR-17-92 is complicated and the results will contribute to further revealing the regulatory mechanisms under the expression of the miR-17-92 cluster.

Isolation and characterization of porcine PILRB gene and its alternative splicing variants.

Paired immunoglobulin-like type 2 receptor (PILR)ß regulates inflammatory responses to pathogen infection, and therefore plays an important role in host disease resistance/susceptibility. However porcine PILRß remains poorly characterized. In this study, we obtained the cDNA (V1) of its encoding gene, PILRB, and three alternative splicing (AS) variants (V2-4). The complete coding sequence of V1 was 621 bp long encoding a polypeptide of 206 aa. Compared with V1, V2 and V3 were formed by exon-skipping in the 3'-untranslated region (UTR), while V4 was formed by alternative 3' splice site of exon 3, resulting in a premature termination codon, combined with exon skipping in the 3'-UTR. Expression profile analysis showed that all the isoforms were most abundant in the spleen, and V1 was strongly induced by poly(I:C). Furthermore, the transcription of V1 altered with the increasing age and differed between species. Exon skipping in the 3'-UTR of V2 and V3 down-regulated expression of the luciferase reporter gene, and hence presumably of the PILRB gene, while V4 was subjected to nonsense-mediated mRNA decay. Additionally, five novel splicing patterns were detected using the minigene approach, indicating complex AS of porcine PILRB. These results will help to reveal the role of PILRß in the host immune response using pig models, and will facilitate the breeding of pigs resistant to viral diseases through molecular breeding methods.

TNFSF9 exerts an inhibitory effect on hepatocellular carcinoma.

OBJECTIVE: Tumor necrosis factor superfamily member 9 (TNFSF9), also known as 4-1BBL and CD137L, has been implicated in cancer immunotherapy due to its function as a T-cell co-stimulator. We aimed to investigate the role of TNFSF9 in the cancer pathogenesis in hepatocellular carcinoma (HCC). METHODS: TNFSF9 expression was examined by immunohistochemistry in 106 pairs of HCC and adjacent non-tumorous tissues, and by quantitative polymerase chain reaction and Western blot in HCC cell lines. The impact of TNFSF9 on the proliferation, migration and invasion of HCC cells was determined using the 3-(4,5-diethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) and transwell assays in vitro. We also assessed the influence of TNFSF9 on the growth and metastasis of HCC tumors in an orthotopic mouse model of human HCC. RESULTS: TNFSF9 expression was downregulated in approximately 70% of HCC tissues. A decreased expression of TNFSF9 was also consistently observed in all the four HCC cell lines. Either the overexpression of TNFSF9 or treatment with recombinant TNFSF9 protein could significantly inhibit the proliferation, migration and invasion of Huh7 and SMMC-7721 HCC cells in vitro. The inhibitory effect of TNFSF9 on HCC was further confirmed in vivo. Mice orthotopically transplanted with TNFSF9-overexpressing Huh7 cells developed significantly smaller tumors with less intrahepatic metastasis and distant metastasis compared with the control group. CONCLUSIONS: TNFSF9 may be a tumor suppressor in HCC. Based on its immune stimulatory aspect and the tumor inhibition property, TNFSF9 may be a promising therapeutic target for HCC.

Carbon Dots/NiCo2 O4 Nanocomposites with Various Morphologies for High Performance Supercapacitors.

A series of carbon dots/NiCo2 O4 composites with various morphologies are prepared and tested for supercapacitors. These samples have good electrical conductivities and efficient ions transport paths, so they exhibit high specific capacitances, superior rate performances, and high cycling stabilities. The optimal composite for hybrid supercapacitor exhibits a high energy density up to 62.0 Wh kg-1 .

[Effects of fractalkine on the expression of inflammatory substances in LPS-activated microglia cells].

AIM: To investigate the effect of fractalkine on the expression of the tumor necrosis factor-α(TNF-α), interleukin-1ß (IL-1ß), and nitric oxide (NO) in LPS-activated murine microglia cells line N9. METHODS: In vitro LPS-activated microglia cells were treated for 24 h in the presence of fractalkine. The level of TNF-α and IL-1ß in the culture supernatants were measured by enzyme-linked immunosorbent assay (ELISA), The level of NO in the culture supernatants were quantitated by the NO test assay. RESULTS: The concentration of TNF-α, IL-1ß and NO in the culture supernatants evidently increased in LPS-activated microglia cells groups and prominently decreased by the fractalkine co-incubated. CONCLUSION: It is thus concluded that fractalkine has neuroprotective functions by inhibiting the expression of inflammatory factor in activated microglia cells.