Ben-Yan Wu

Foxp3+IL‐17+ T cells promote development of cancer‐initiating cells in colorectal cancer

The pathogenesis of CRC remains to be further understood. This study was designed to elucidate the role of Foxp3+IL‐17+ T cells in the pathogenesis of CRC. Surgically removed CRC tissue was collected from 12 patients with CRC. The frequency and cytokine profile of Foxp3+IL‐17+ T cells in CRC were examined by flow cytometry. Chemokine CXCL11 was examined in CRC tissue by Western blotting. Treg chemotaxis was examined in a transwell system. The effect of Foxp3+IL‐17+ T cells on induction of cancer‐initiating cells was examined; the latterˈs Akt and MAPK activities and colony formation were examined afterward. Abundant Foxp3+IL‐17+ T cells were detected in CRC tissue that expresses high levels of TGF‐β, CXCR3, CCR6, and RORγt. High levels of CXCL11 were detected in CRC tissue‐derived CD68+ cells, which had a strong chemotactic effect on Foxp3+ Tregs. Hypoxia induced the expression of IL‐17 in Foxp3+ Tregs; Foxp3+IL‐17+ T cells were capable of inducing CRC‐associated cell markers in BMMo and drove the cells to be cancer‐initiating cells. High levels of phosphorylated Akt and MAPK were detected in the induced cancer‐initiation cells; the latter has the capability to form a colony. CRC tissue‐derived Foxp3+IL‐17+ cells have the capacity to induce cancer‐initiating cells.

Oxymatrine liposome attenuates hepatic fibrosis via targeting hepatic stellate cells

AIM To investigate the potential mechanism of Arg-Gly-Asp (RGD) peptide-labeled liposome loading oxymatrine (OM) therapy in CCl₄-induced hepatic fibrosis in rats. METHODS We constructed a rat model of CCl₄-induced hepatic fibrosis and treated the rats with different formulations of OM. To evaluate the antifibrotic effect of OM, we detected levels of alkaline phosphatase, hepatic histopathology (hematoxylin and eosin stain and Masson staining) and fibrosis-related gene expression of matrix metallopeptidase (MMP)-2, tissue inhibitor of metalloproteinase (TIMP)-1 as well as type I procollagen via quantitative real-time polymerase chain reaction. To detect cell viability and apoptosis of hepatic stellate cells (HSCs), we performed 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-diphenytetrazoliumromide assay and flow cytometry. To reinforce the combination of oxymatrine with HSCs, we constructed fluorescein-isothiocyanate-conjugated Arg-Gly-Asp peptide-labeled liposomes loading OM, and its targeting of HSCs was examined by fluorescent microscopy. RESULTS OM attenuated CCl₄-induced hepatic fibrosis, as defined by reducing serum alkaline phosphatase (344.47 ± 27.52 U/L vs 550.69 ± 43.78 U/L, P < 0.05), attenuating liver injury and improving collagen deposits (2.36% ± 0.09% vs 7.70% ± 0.60%, P < 0.05) and downregulating fibrosis-related gene expression, that is, MMP-2, TIMP-1 and type I procollagen (P < 0.05). OM inhibited cell viability and induced apoptosis of HSCs in vitro. RGD promoted OM targeting of HSCs and enhanced the therapeutic effect of OM in terms of serum alkaline phosphatase (272.51 ± 19.55 U/L vs 344.47 ± 27.52 U/L, P < 0.05), liver injury, collagen deposits (0.26% ± 0.09% vs 2.36% ± 0.09%, P < 0.05) and downregulating fibrosis-related gene expression, that is, MMP-2, TIMP-1 and type I procollagen (P < 0.05). Moreover, in vitro assay demonstrated that RGD enhanced the effect of OM on HSC viability and apoptosis. CONCLUSION OM attenuated hepatic fibrosis by inhibiting viability and inducing apoptosis of HSCs. The RGD-labeled formulation enhanced the targeting efficiency for HSCs and the therapeutic effect.

Recurrence in patients following curative resection of early gastric carcinoma

Post‐gastrectomy recurrences of early gastric cancer occur in a few cases. We investigated the outcome of early gastric cancer patients treated surgically, with special respect to the risk factor(s) for tumor recurrence.

Immunoglobulin kappa and immunoglobulin lambda are required for expression of the anti-apoptotic molecule Bcl-xL in human colorectal cancer tissue

Abstract Objective. Aberrant expression of immunoglobulin (Ig) by cancer cells has been documented in a number of malignant tumors but its biological significance is unclear. Cancer cells overexpress anti-apoptotic molecules such as Bcl-xL. The present study aimed to examine the role of expression of Ig light-chain Igĸ and Igλ in maintaining the high levels of Bcl-xL in colorectal cancer cells. Material and methods. Thirty patients with colorectal cancer were recruited to this study. Expression of Igĸ, Igλ and Bcl-xL in surgically removed cancer tissue was examined by immunohistochemistry and/or flow cytometry. Using the HT29 cell line as a study platform, RNA interference (RNAi) was employed to knock out the genes of Igĸ and Igλ in the cancer cell line; the expression of Bcl-xL in HT29 cells was subsequently analyzed. Results. Human colorectal cancer cells, but not normal colorectal tissue, expressed both Igĸ and Igλ in the cytoplasm. High levels of Bcl-xL were detected in cancer cells. Using RNAi to knock out the genes of Igĸ and/or Igλ, Bcl-xL expression in HT29 cells was significantly suppressed and the cells became apoptotic. Conclusion. The results suggest that expression of Igĸ and Igλ is required to stabilize Bcl-xL expression in cancer cells.

Suppression of CB1 cannabinoid receptor by lentivirus mediated small interfering RNA ameliorates hepatic fibrosis in rats.

It is recognized that endogenous cannabinoids, which signal through CB1 receptors in hepatic stellate cells (HSCs), exert a profibrotic effect on chronic liver diseases. In this study, we suppressed CB1 expression by lentivirus mediated small interfering RNA (CB1-RNAi-LV) and investigated its effect on hepatic fibrosis in vitro and in vivo. Our results demonstrated that CB1-RNAi-LV significantly inhibited CB1 expression, and suppressed proliferation and extracellular matrix production in HSCs. Furthermore, CB1-RNAi-LV ameliorated dimethylnitrosamine induced hepatic fibrosis markedly, which was associated with the decreased expression of mesenchymal cell markers smooth muscle α-actin, vimentin and snail, and the increased expression of epithelial cell marker E-cadherin. The mechanism lies on the blockage of Smad signaling transduction induced by transforming growth factor β1 and its receptor TGF-β RII. Our study firstly provides the evidence that CB1-RNAi-LV might ameliorate hepatic fibrosis through the reversal of epithelial-to-mesenchymal transition (EMT), while the CB1 antagonists AM251 had no effect on epithelial-mesenchymal transitions of HSCs. This suggests that CB1 is implicated in hepatic fibrosis and selective suppression of CB1 by small interfering RNA may present a powerful tool for hepatic fibrosis treatment.

Integrin alphavbeta6 promotes tumor tolerance in colorectal cancer

Tumor immune tolerance plays a critical role in tumor cell survival; the establishment of tumor immune tolerance is incompletely understood yet. Integrin alphavbeta6 (avb6) is involved in tumor growth and metastasis. This study aimed to observe the effect of avb6 on the development of tumor tolerance in colorectal cancer (CRC). In this study, 28 CRC patients were recruited. The frequencies of tolerogenic dendritic cells (TolDC), regulatory T cells (Treg), and CD8+ T cells in surgically removed CRC tissue were assessed by flow cytometry. The levels of avb6 in CRC tissue were measured by enzyme-linked immunoassay (ELISA). The effect of avb6 on inducing TolDCs and Tregs was evaluated with the cell culture model. The results showed that in surgically removed CRC tissue, we detected higher frequencies of TolDC and Tregs, lower frequency CD8+ T cells and high levels of avb6 as compared with non-CRC tissue. CRC protein extracts could induce TolDC development that could be blocked by anti-avb6 antibody. CRC-derived DCs could convert naïve CD4+ T cells to Tregs. Peripheral CD8+ T cells from CRC patients still retained the ability to produce granzyme B and to proliferate in response to CRC tumor antigen in culture that was abolished by the presence of CRC-derived Tregs. We conclude that CRC-derived avb6 is involved in the establishment of tumor immune tolerance in local tissues.

THYROID HORMONES PRECIPITATE SUBCLINICAL HYPOPITUITARISM RESULTED IN ADRENAL CRISIS

ing the 6 years of follow-up, 44 died (20 with LTCI certification and 24 without). The combined rate of incident disability or death was lower in the categories with low NT-pro BNP and higher in the categories with high NT-pro BNP (Q1, 37.9/1,000 person-years; Q5, 115.8/1,000 person-years; Table 1). This relationship was also evident in the ageand sex-adjusted model. After adjustment for possible confounding factors, the HR of incident disability or death was significantly higher in Q5 (NT-pro BNP 241 pg/mL) than in Q1. These results persisted when systolic BP values and antihypertensive medication instead of BP categories were included as confounding factors. When incident disability was used only as an endpoint, the risk was statistically significantly greater in group Q5 than in Q1. Thus, the present study confirms that plasma NT-pro BNP is positively associated with incident disability and death. This relationship was independent of physical function and other possible confounders.