Clophosome alleviate dextran sulphate sodium-induced colitis by regulating gut immune responses and maintaining intestinal integrity in mice.

Inflammatory bowel disease (IBD) is a chronic progressive disorder characterized by complicated gastrointestinal inflammation. Research on therapeutic agents is still urgent due to the lack of satisfactory treatments. Gut macrophages are considered to be predominant in excessive inflammatory responses. Thus, we aimed to investigate whether depletion of macrophages would have a beneficial effect on IBD and could be a potential therapeutic strategy. In this study, we established a 12-day Dextran sodium sulphate (DSS)-induced colitis mouse model and determined the effect of the macrophage depletion agent Clophosome (neutral clodronate liposomes; CNC). The results showed that CNC significantly alleviated the symptoms of colitis, as demonstrated by greater weight gain, decreased disease activity index (DAI) scores, and lower histopathological damage scores, as well was reduced levels of the proinflammatory cytokines interleukin (IL)-6 and tumour necrosis factor (TNF)-α. To investigate T cell subsets, cells were isolated from the lamina propria and cultured to analyse the expression of IL-17A, interferon (IFN)-γ and Foxp3 in CD4+ cells by flow cytometry. The data showed that during the process of colitis, the frequencies of CD4+ IL-17A+ T cells were significantly increased. Notably, CNC treatment markedly reduced the population of CD4+ IL-17A+ T cells, especially CD4+ IL-17A+ IFN-γ+ T cells. Furthermore, intestinal barrier integrity, as assessed by immunostaining of mucin and tight junction proteins, was severely disrupted in colitis. CNC improved the intestinal barrier by enhancing the expression of muc-2 and occludin. In summary, our findings demonstrated that CNC successfully ameliorated DSS-induced colitis and that its effect may be associated with inhibiting inflammatory responses and maintaining intestinal integrity.

[Clinical characteristics and survival of peripheral T-cell lymphoma-not otherwise specified: a report of 57 cases].

OBJECTIVE: To analyze the clinical characteristics and survival data of 57 patients with peripheral T-cell lymphoma-not otherwise specified (PTCL-NOS). METHODS: The medical records of 57 patients with PTCL-NOS classified according to the revised REAL-WHO criteria, treated from Jan 1993 to Dec 2007 at the First and the Third affiliated Hospitals of Medical School of Xi'an Jiaotong University, were retrospectively evaluated by K-M univariate and COX multivariate analysis. RESULTS: 39 of the patients (68.4%) were males and 18 (31.6%) were females, aged 44 (3 - 88). Nine of the 57 patients (15.8%) were treated with chemo-radiotherapy, 43 (75.4%) with chemotherapy, 3 (5.3%) with radiotherapy, and 2 with supported treatment alone (2.5%). The overall response rate was 87.3%, with a complete remission (CR) rate of 60.0% in 55 evaluable cases. The 1-, 3-, and 5-year overall survival (OS) rates were 67.0%, 48.0% and 24.3%, respectively, with a median follow-up of 30.4 months (ranged 1-100 months). The median survival time (MST) was 36.0 months. Multivariate analysis showed that the prognostic index for T cell lymphoma (PIT) score was an independent prognostic factor for PTCL-NOS (P < 0.05), but bone marrow involvement, performance status, extranodal involvement, stage, B symptom were not independent prognostic factors. CONCLUSION: Although conventional chemotherapy yields a high response rate for PTCL-NOS, the long-term survival is still low and further investigation for optional treatment is needed.

[The relationship between lymphatic metastasis and serum vascular endothelial growth factor C and cyclooxygenase 2 expression in breast cancer].

OBJECTIVE: To investigate the relationship among the serum vascular endothelial growth factor C (sVEGF-C), expression of cyclooxygenase 2 (COX-2), and lymph vessel density (LMVD), and to discuss their role in tumor progression and lymphatic metastasis. METHODS: sVEGF-C level was detected by ELISA in 68 pre-operation breast cancer patients, 35 breast benign disease patients, and 12 healthy women. Immunohistochemical method was used to detect the expression of COX-2 and lymphatic vessel endothelial hyaluronidase receptor (LYVE)-1 in the breast cancer tissues and benign disease tissues obtained during operation. RESULTS: (1) The sVEGF-C level of the pre-operation breast cancer group was (49 +/- 8) pg/ml, significantly higher than those of the begin tumor group [(8 +/- 4) pg/ml, P = 0.045] and the healthy women [(5 +/- 3) pg/ml, P = 0.003]. (2) The COX-2 overexpression positive rate in the breast cancer tissues was 44.1%, significantly higher than that in the begin disease tissues (11.4%, P = 0.002). Higher than that in the begin disease tissue; the LMVD overexpression in the breast cancer tissues was mainly located in the para-carcinoma tissue (8.7 +/- 4.7), significantly higher than that in the begin disease tissues (1.8 +/- 1.7, P = 0.002). (3) The COX-2 overexpression rate of the patients with a high sVEGF-C level was significantly higher than that of the patients with a low sVEGF-C level (P = 0.024). The LMVD level in the breast cancer of the sVEGF-C high level group was (8.7 +/- 3.9), significantly higher than that of the sVEGF-C normal group (5.6 +/- 3.3, P = 0.039). (4) The LMVD in the COX-2 overexpression breast cancer group was 9.5 +/- 3.7, significantly higher than that in the COX-2 negative group (6.0 +/- 3.8, P = 0.012). (5) The sVEGF-C level, COX-2, and LMVD expression in the breast cancer patients with lymphatic metastasis were significantly higher than those in the patients without lymphatic metastasis (P = 0.025, 0.022, and 0.002 respectively). CONCLUSION: VEGF-C, COX-2, and LMVD play important roles in the lymphatic metastasis of breast cancer. They may be important prognostic factors in evaluating breast cancer lymphatic metastasis.

[Relationship between serum VEGF level and VEGF, COX-2 and MVD expression in breast cancer tissues].

OBJECTIVE: To investigate the relationship between serum VEGF (sVEGF) level and VEGF, COX-2 and MVD expression in breast cancer, and to discuss their role in angiogensis of breast cancer. METHODS: sVEGF level was detected by ELISA in 68 preoperative breast cancer, 35 benign breast disease and 20 healthy women. The expression of VEGF, COX-2 and MVD was detected by immunohistochemical method in tissues of breast cancer and breast benign diseases, and to analyze the relationship of sVEGF, VEGF, COX-2 and MVD. RESULTS: (1) sVEGF level in preoperative breast cancers was 306.51 pg/ml (interquartile range from 190.44 to 442.04 pg/ml), in benign diseases was 150.82 pg/ml (interquartile range from 82.36 to 212.34 pg/ml), and in healthy control was 105.93 pg/ml (interquartile range from 78.54 to 157.77 pg/ml). The sVEGF level of preoperative breast cancer group was significantly higher than that of breast benign disease group and healthy women (P = 0.001). (2) The VEGF expression positive rate in breast cancer (67.65%) was significantly higher than that in breast benign disease (44.12%) (P = 0.015). The COX-2 expression positive rate in breast cancer (42.86%) was significantly higher than that in breast benign disease (11.43%) (P = 0.002). (3) the COX-2 expression positive rate in sVEGF high level patients (56.00%) was significantly higher than that in sVEGF normal level patients (11.11%) (P = 0.024), and MVD in sVEGF high level patients (27.32 +/- 3.40) was also higher than that in sVEGF normal level patients (15.31 +/- 6.16) (P = 0.011). (4) The sVEGF level (322.09 +/- 79.31) of 68 breast cancer patients whose VEGF was positive in breast cancer tissues was significantly higher than that in VEGF negative group (222.47 +/- 73.53) (P = 0.017). (5) The COX-2 expression positive rate in VEGF positive expression group (65.21%) was significantly higher than that in VEGF negative expression group (18.18%) (P = 0.017). The MVD expression in COX-2 positive expression group (22.94 +/- 5.51) was significantly higher than that in COX-2 negative expression group (10.30 +/- 4.42) (P = 0.027). CONCLUSION: sVEGF level in breast cancer is significantly higher than that in breast benign disease and healthy women, and is correlated with the expression of COX-2 and MVD in breast cancer tissues.

Characteristics of Differently Located Colorectal Cancers Support Proximal and Distal Classification: A Population-Based Study of 57,847 Patients.

BACKGROUND: It has been suggested that colorectal cancer be regarded as several subgroups defined according to tumor location rather than as a single entity. The current study aimed to identify the most useful method for grouping colorectal cancer by tumor location according to both baseline and survival characteristics. METHODS: Cases of pathologically confirmed colorectal adenocarcinoma diagnosed from 2000 to 2012 were identified from the Surveillance, Epidemiology, and End Results database and categorized into three groups: right colon cancer (RCC), left colon cancer (LCC), and rectal cancer (ReC). Adjusted hazard ratios for known predictors of disease-specific survival (DSS) in colorectal cancer were obtained using a Cox proportional hazards regression model. RESULTS: The study included 57847 patients: 43.5% with RCC, 37.7% with LCC, and 18.8% with ReC. Compared with LCC and ReC, RCC was more likely to affect old patients and women, and to be at advanced stage, poorly differentiated or un-differentiated, and mucinous. Patients with LCC or ReC had better DSS than those with RCC in subgroups including stage III or IV disease, age ≤70 years and non-mucinous adenocarcinoma. Conversely, patients with LCC or ReC had worse DSS than those with RCC in subgroups including age ˃70 years and mucinous adenocarcinoma. CONCLUSIONS: RCC differed from both LCC and ReC in several clinicopathologic characteristics and in DSS. It seems reasonable to group colorectal cancer into right-sided (i.e., proximal) and left-sided (i.e., distal) ones.

IDH1 overexpression induced chemotherapy resistance and IDH1 mutation enhanced chemotherapy sensitivity in Glioma cells in vitro and in vivo.

Isocitrate dehydrogenase (IDH) is of great importance in cell metabolism and energy conversion. IDH mutation in glioma cells is reported to be associated with an increased overall survival. However, effects biological behavior of therapy of gliomas are unclear. Here, we investigated the influence of wild-type and mutated IDH genes on glioma cell biological behavior and response to chemotherapy. Relevant mechanisms were further explored. We designed our study on the background of the IDHR132H mutation. Stable cell lines were constructed by transfection. The CCK-8 method was used to assess cell proliferation, flow cytometry for the cell cycle and cell apoptosis, and the transwell method for cell invasion. Nude mouse models were employed to determine tumorigenesis and sensitivity to chemotherapy. Western blotting was used to detect relevant protein expression levels. We found that overexpression of wild IDH1 gene did not cause changes in the cell cycle, apoptosis and invasion ability. However, it resulted in chemotherapy resistance to a high dose of temozolomide (TMZ) in vivo and in vitro. The IDH1 mutation caused cell cycle arrest in G1 stage and a reduction of proliferation and invasion ability, while raising sensitivity to chemotherapy. This may provide an explanation for the better prognosis of IDH1 mutated glioma patients and the relative worse prognosis of their wild-type IDH1 counterparts. We also expect IDH1 mutations may be optimized as new targets to improve the prognosis of glioma patients.

Mechanisms underlying the biological changes induced by isocitrate dehydrogenase-1 mutation in glioma cells.

Isocitrate dehydrogenase 1 (IDH1) mutation has been reported to be associated with an increased overall survival in patients with glioma in a number of studies. Previous studies have focused on the mutation rate and possible metabolic pathways of the mutated IDH1 gene. However, the effects of IDH1 mutation on the biological behavior of glioma cells and the associated mechanisms, as well as the possible effects they may have on clinical therapy, have not been studied. In the present study, three eukaryotic expression vectors were constructed and transfected into the U87 cell line, specifically, a wild-type form of the IDH1 gene with the enhanced green fluorescent protein (EGFP) gene, a mutated IDH1 gene with the EGFP gene and the EGFP gene only. The three stable cell lines were selected using the G418 antibiotic. The biological behaviors of the cell lines were studied and the mechanisms underlying the biological differences between the cell lines were further investigated. The present study confirmed that IDH1 mutation induced cell cycle arrest in the G1 phase and reduced the proportion of the G2/M phase, by downregulating cell division control protein 2 homolog levels, increasing bromodomain-containing protein 2 levels and markedly limiting cell proliferation. IDH1 mutation had no effect on the apoptosis rate under routine culture conditions. Serum chemotaxis assays showed that IDH1 mutation was markedly associated with a significantly reduced invasion ability, by reducing the levels of matrix metalloproteinase (MMP)-2 and MMP-9. From this study, it may be concluded that IDH1 mutation improves prognosis in glioma patients by altering the cell cycle, inhibiting cell proliferation and downregulating cell invasion ability. The results may provide a partial explanation for the improved prognosis of patients with mutated forms of the IDH1 gene.

Induction of p53 expression and apoptosis by a recombinant dual-target MDM2/MDMX inhibitory protein in wild-type p53 breast cancer cells.

The tumor suppressor gene p53 is often inactivated in breast cancer cells due to gene mutation or overexpression of its repressors (such as murine double minute 2 and murine double minute X). Inhibitors of murine double minute 2 (MDM2) and murine double minute X (MDMX) could lead to tumor suppression by restoration of p53 activity and such an approach is a promising strategy for future control of breast cancer. This study aimed to investigate the feasibility of the recombinant MDM2 and MDMX inhibitory protein in control of breast cancer in vitro. A cell-permeable dual-target MDM2/MDMX inhibitory protein was expressed in E. coli and incubated with p53 wild-type breast cancer cells. The data showed that this recombinant MDM2/MDMX inhibitory protein reduced the viability of MCF-7 and ZR-75-30 breast cancer cell lines and promoted cell cycle arrest and apoptosis by activation and stabilization of the p53 protein. Mechanistically, this MDM2/MDMX inhibitory protein increased the expression of p21, Bax and puma proteins, and inhibitory expression of MDM2 and MDMX proteins. This recombinant protein showed a better in vitro effect than that of nutlin-3α, a small molecule MDM2 inhibitor. The data further support the hypothesis that targeting of the p53 gene pathway could effectively control breast cancer.

Advances and prospects of anginex as a promising anti-angiogenesis and anti-tumor agent.

Anginex, a novel artificial cytokine-like peptide (ßpep-25), is designed by using basic folding principles and incorporating short sequences from the ß-sheet domains of anti-angiogenic agents, including platelet factor-4 (PF4), interleukin-8 (IL-8), and bactericidal-permeability increasing protein 1 (BP1). Anginex can specially block the adhesion and migration of the angiogenically activated endothelial cells (ECs), leading to apoptosis and ultimately to the inhibition of angiogenesis and tumor growth. In vitro and in vivo studies have proved its inhibitory effects on the formation of new blood vessels and tumor growth even though the mechanism is not clear. The inhibitory effects of anginex can be enhanced when it is applied in combination with other therapies, such as chemotherapy, radiotherapy and other anti-angiogenic agents. The limitations of anginex, including poor stability, short half life, complicated synthesis and low purity, have been conquered by modifying its structure or designing novel compound anginex and recombinant anginex, which makes possible the clinical application of anginex. Here, we summarize the basic and preclinical trials of anginex and discuss the prospects of anginex in clinical application. We come to the conclusion that anginex and compound or recombinant anginex can be used as effective anti-angiogenic agents.

Anti-angiogenesis and anti-tumor effects of AdNT4-anginex.

Anginex is a novel artificial peptide that can inhibit angiogenesis. AdNT4-anginex was constructed by inserting the artificial anginex gene into a recombinant adenoviral vector. We demonstrated that AdNT4-anginex inhibited migration of human endothelial cells, angiogenesis and tumor growth in in vitro and in vivo studies. Tumor growth of human H22 hepatoma in mice was inhibited after AdNT4-anginex treatment for 4 weeks, and a significant decrease in tumor size was observed as compared with the control group. Overall, these studies indicate that AdNT4-anginex is an effective anti-tumor agent, and deserves more attention and research.