ARPC1B is a novel prognostic biomarker for kidney renal clear cell carcinoma and correlates with immune infiltration.

Background: Actin-related protein 2/3 complex subunit 1B (ARPC1B) is reported to be involved in tumorigenesis and progression. However, its role in kidney renal clear cell carcinoma (KIRC), correlation with tumor-infiltrating immune cells, and prognostic significance remain unclear. Methods: Data sets from the TCGA, GTEx, GEPIA, GEO, UALCAN, and CPTAC databases were extracted and analyzed to investigate the expression difference, prognosis, and clinicopathological features of ARPC1B. Single-sample Gene Set Enrichment Analysis (ssGSEA), CIBERSORT, and TISCH2 analysis were used to examine the relationship between ARPC1B expression and tumor immune infiltration in KIRC. The potential function of ARPC1B in KIRC was explored by GO functional annotation and KEGG pathway analysis. The TIDE algorithm was used to predict and analyze the relationship between ARPC1B expression and response to immune checkpoint blockade (ICB). The expression of ARPC1B was further validated by using quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC). Results: The study showed that ARPC1B expression was an independent prognostic factor of KIRC, with high ARPC1B expression being associated with poor overall survival (OS). Enrichment of GO annotation and pathway analysis showed multiple immune-related functional pathways affected by ARPC1B such as regulation of immune effector process, inflammatory response regulation, antigen processing and presentation, asthma, autoimmune thyroid disease, graft versus host disease, intestinal immune network for IgA production, and type I diabetic mellitus. Moreover, ARPC1B expression positively correlated with infiltrating levels of myeloid-derived suppressor cells (MDSCs) and regulatory T cells (Tregs) in KIRC. Importantly, high ARPC1B expression predicted a low response to ICB in KIRC. Conclusion: This study indicates that ARPC1B expression is an independent prognostic biomarker for OS in KIRC patients. High ARPC1B expression is closely associated with MDSCs and Tregs infiltration. These findings suggest that ARPC1B may serve as a biomarker for prognosis and immune infiltration in KIRC, potentially aiding in the development of novel treatment strategies to improve the survival outcomes for KIRC patients.

Landscape pattern evolution along terrain gradient in Fuzhou City, Fujian Province, China.

Terrain is an important factor in land landscape pattern change. To reveal the spatial and temporal characteristics and variation of landscape pattern along a terrain gradient, we used three remote sensing images and digital elevation model (DEM) data of Fuzhou in 1995, 2005 and 2015 to investigate the topographic gradient effect of landscape pattern and the cause of formation based on terrain index, land use distribution index, geo-informatics map analysis and landscape index. The results showed that forestland was mainly distributed in middle-low, middle-high and higher level terrain, while farmland, water body, construction land and unused lands were mainly distributed in the low rank terrain. From 1995 to 2015, the area of forestland, farmland, and unused land in Fuzhou decreased, while that of construction land and water body increased. The change of landscape type was stable, which was mainly distributed in middle-low, middle-high, and high grade terrain gradient region. In addition, landscape pattern changes were obvious across the terrain gra-dient. The landscape type of low terrain area mainly shifted to the construction land, but farmland and forestland had an alternation change in the middle-low and middle-high terrain area. The characteristics of landscape pattern fragmentation, landscape heterogeneity and landscape diversity in the study area had been increasing year by year, but they had decreased with the elevation of terrain.

1810011o10 Rik Inhibits the Antitumor Effect of Intratumoral CD8+ T Cells through Suppression of Notch2 Pathway in a Murine Hepatocellular Carcinoma Model.

The mechanisms by which tumor-responsive CD8+ T cells are regulated are important for understanding the tumor immunity and for developing new therapeutic strategies. In current study, we identified the expression of 1810011o10 Rik, which is the homolog of human thyroid cancer 1, in intratumoral activated CD8+ T cells in a murine hepatocellular carcinoma (HCC) implantation model. To investigate the role of 1810011o10 Rik in the regulation of antitumor activity of CD8+ T cells, normal CD8+ T cells were transduced with 1810011o10 Rik-expressing lentiviruses. Although 1810011o10 Rik overexpression did not influence agonistic antibody-induced CD8+ T cell activation in vitro, it inhibited the cytotoxic efficacy of CD8+ T cells on HCC cells in vivo. 1810011o10 Rik overexpression impeded CD8+ T cell-mediated HCC cell apoptosis and favored tumor cell growth in vivo. Further investigation revealed that 1810011o10 Rik blocked the nuclear translocation of Notch2 intracellular domain, which is crucial for CD8+ T cell activity. Furthermore, a brief in vitro experiment suggested that both antigen-presenting cells and TGF-ß might be necessary for the upregulation of Rik expression in activated CD8+ T cells. In general, our study disclosed a novel mechanism underlying the negative regulation of antitumor CD8+ T cells during HCC progression.

Myxoid adrenocortical adenoma with a pseudoglandular pattern: a case report and literature review.

Adrenocortical adenoma is a benign neoplasm derived from cells of the adrenal cortex. The myxoid variant of this tumor is extremely rare. To our knowledge, only 23 cases of myxoid adrenocortical adenoma have been reported so far and 19 of them mentioned the pseudoglandular pattern. We reported a new case of 56-year-old Chinese female patient whose left adrenal gland was shown a neoplastic lesion by computed tomography (CT) and magnetic resonance (MR) imaging. Histopathological study showed that the mass was a myxoid adrenocortical adenoma with a pseudoglandular pattern. Then, we performed immunohistochemistry with 28 biomarkers to make differential diagnosis and found that tumor cells were diffusely positive for vimentin, melan-A, CD56, NSE and USP10, and focally positive for cytokeratin pan, cytokeratin 8/18 and VEGF. The labeling index of Ki-67 and Cyclin D1 were about 1% and 50%, respectively. No immunoreactivity was found for EMA, cytokeratin 7, HMB45, S-100, alpha-inhibin, calretinin, synaptophysin, chromogranin A, P53, EGFR, MMP2, DNA topo II alpha, CA125, E-cadherin, P63, P16 and Her-2. The patient has been followed up for 37 months after tumor resection and no evidence was found to suggest any local recurrence or any metastatic disease. Myxoid adrenocortical adenoma with a pseudoglandular pattern is extremely rare. The accurate diagnosis should be based on combined consideration of clinical characteristics, CT, MR imaging and pathological features, and should be distinguished from other retroperitoneal myxoid tumors.

Prognostic significance of USP10 as a tumor-associated marker in gastric carcinoma.

Ubiquitin-specific protease 10 (USP10), a novel deubiquitinating enzyme, had been associated with growth of tumor cell. However, the role of USP10 in gastric cancer carcinogenesis had not been elucidated yet. The aim of this study was to investigate the expression level of USP10 in gastric carcinoma (GC) tissues and cell lines, then to evaluate the clinical significance of USP10 in GC patients. USP10, E-cadherin, Ki67 and p53 expressions were detected in 365 GC and 40 non-cancerous mucosa tissues by immunohistochemistry. Western blot for USP10 was performed on additional fresh GC tissues and GC cell lines. The expression level of USP10 in GC tissues was proved lower than that in non-cancerous mucosa tissues (p < 0.05). It was also lower in GC cell lines (AGS, BGC-823 and MKN45 cells) than that in gastric epithelial immortalized cell line (GES-1). Clinicopathological analysis showed that USP10 expression was negatively correlated with gastric wall invasion (p = 0.009), nodal metastasis (p = 0.002), and TNM stage (p = 0.000). In contrast, a positively correlation between the expression of USP10 and E-cadherin was found (p < 0.05), but there was no relationship proved between Ki67, p53 and USP10 (p > 0.05). On the Kaplan-Meier survival curves, we found poor prognosis in GC patients was associated with negative USP10 expression (p < 0.05). Moreover, USP10 expression was an independent prognostic factor for the overall survival in multivariate analysis. Our findings suggested that USP10 was an independent predictor of prognosis of GC patients.

[The research of xeno-lymphocytes apoptosis induced by Sertoli cells isolated by modified methods in vitro].

OBJECTIVE: To modify the isolation and culture method of Sertoli cells and investigate its' effects on xeno-lymphocytes apoptosis. METHODS: Sertoli cells which was isolated from 2 - 4 week-old Sprague Dawley (SD) rats, were successfully prepared by collagenase type V, trypsin and DNase I and then identified by electron microscope. Viability and apoptosis of cultured cells were measured by flow cytometry. The apoptosis rates of Balb/c mouse lymphocytes were examined which were co-cultured with Sertoli cells of SD rats by flow cytometry, too. The expression of FasL, TGF-beta(1) and clusterin on Sertoli cells were detected by immunocytochemistry. RESULTS: In the co-cultured system, Sertoli cells accounted for more than 90%. The viability of Sertoli cells was above 95% and the apoptosis rate was 10.87% +/- 3.87% in this study. The lymphocytes apoptosis ratio was 15.52% +/- 0.17% (P < 0.01). Streptavidin-biotin-peroxidase-complex immunochemistry staining showed that the Sertoli cells could express FasL, TGF-beta(1) and clusterin, respectively. CONCLUSIONS: It indicates that the expression of FasL, TGF-beta(1) on the Sertoli cells might relate to the immune privilege, and it supposed to be benefit for xenotransplantation.

[Coexpression of human complement regulatory protein genes, CD59 and MCP in transgenic mice protects isolated mouse heart function in ex vivo human plasma perfusion model].

OBJECTIVE: To investigate if the expression of human complement regulatory protein genes in transgenic donor protects against hyperacute rejection (HAR) in the recipient. METHODS: Kunming mice were transfected with 2 human complement regulatory protein genes, CD59 and MCP, so as to establish a transgenic hCD59/hMCP mouse model. Eight hCD59 expression mice, 11 hMCP expression mice, and 8 hCD59/hMCP expression mice were used as experimental groups, and 10 transgenic negative littermates were used as control group. The hearts of the mice were taken out to be perfused with 10% pooled human blood of B type. During the perfusion electrocardiography was carried out to observe the beating time. After the hearts stopped beating, immunofluorescence staining and immunohistochemistry were used to detect the deposition of complements C(9) and C(3c) in the heart tissue. RESULTS: The mean heart beating time was 138 +/- 25 minutes in the hCD59/hMCP expression group, 78 +/- 27 minutes in the hCD59 expression group, 43 +/- 21 minutes in the hMCP group, and 20 +/- 12 minutes in the wild type nontransgenic control group (all P < 0.01, Dennett's T test for all 3 other groups relative to the hCD59/hMCP group). Deposition of the complement C(9) and that of C(3c) were not found in the hearts of the hCD59/hMCP group, however, could be found in the hearts of the 2 monotransgenic groups and nontransgenic group. CONCLUSION: The coexpression of human complement regulatory protein genes, CD59 and MCP in the xenografts effectively inhibits the complement of xenograft-mediated HAR.

Inhibition of xenogeneic response in porcine endothelium using RNA interference.

BACKGROUND: Rejection mediated by antibody recognition of the alpha-Gal epitope (Galalpha1-3Galbeta1-4GlcNAc-R) is a major barrier in porcine-to-human xenotransplantation. Because the synthesis of alpha-Gal is dependent on alpha1,3 galactosyltransferase (alpha1,3GT), methods of blocking this enzyme are needed. RNA interference induced by small interfering RNA (siRNA) is a powerful technique for allowing the silencing of mammalian genes with great specificity and potency. In this study, we use siRNA for silencing of alpha1,3GT with the purpose of reducing expression of the alpha-Gal epitope and subsequently decreasing immunogenicity of porcine endothelial cells. METHODS: alpha1,3GT-specific and control siRNAs were transfected into the porcine aortic endothelial cell line, PED. alpha-Gal expression was assessed by Western blotting, flow cytometry, and immunofluorescence. Protection from human-complement and natural killer (NK)-cell-mediated cytotoxicity was evaluated by Cr-release assays after incubation of PED with normal human serum (NHS) and NK92 cell, respectively. RESULTS: RNA interference was successfully achieved in PED as witnessed by the specific knock-down of alpha1,3GT mRNA levels. Flow cytometric analysis using the Griffonia simplicifolia isolectin B4 lectin confirmed the suppression of alpha1,3GT activity as evidenced by decreased alpha-Gal. Functional relevance of the knock-down phenotype was illustrated by the finding that silenced PED were protected from cytotoxicity of NHS. Protection from NK-mediated cytotoxicity was not observed. CONCLUSIONS: Our data are the first to demonstrate that RNA interference is a potent tool to down modulate alpha-Gal expression and to protect endothelial cells from complement-mediated cytotoxicity. Gene silencing by siRNA may represent a new approach for overcoming hyperacute and acute vascular rejection.