Notch signaling pathway regulates T cell dysfunction in septic patients.

Sepsis disrupts innate and adaptive immune response, and immune disorders may also impact clinical course of sepsis. Notch signaling pathway plays a vital role in T cell modulation and differentiation. The aim of current study was to investigate the immunoregulatory function of Notch signaling pathway to T cells in patients with sepsis and septic shock. Twenty-seven sepsis patients, twenty-five septic shock patients, and twenty-one normal controls (NCs) were enrolled. Notch receptors mRNA levels were semi-quantified by real-time PCR. The absolute numbers of CD3+, CD4+, and CD8+ T cells were measured by flow cytometry. Key transcriptional factors of CD4+ T cells, cytotoxic molecules in CD8+ T cells, and cytotoxicity of CD8+ T cells were investigated. The regulatory activities of Notch signaling inhibition by γ-secretase inhibitor (GSI) on purified CD4+ and CD8+ T cells from sepsis and septic shock patients were also assessed. Notch1 mRNA relative level was significantly elevated in sepsis and septic shock patients when compared with NCs. CD4+ and CD8+ T cells were dysfunctional in sepsis and septic shock, which presented as decreased cell accounts, down-regulation of Th1/Th17 transcriptional factors and cytotoxic molecules (perforin, granzyme B, and FasL), and reduced cytotoxicity of CD8+ T cells. Notch signaling inhibition by GSI increased Th1 and Th17 differentiation of CD4+ T cells. Moreover, GSI stimulation not only promoted perforin, granzyme B, and FasL mRNA expression in CD8+ T cells, but also elevated CD8+ T cell-induced target cell death and IFN-γ/TNF-α production in sepsis and septic shock. The current data suggest that Notch signaling pathway contributes to T cell dysfunction and limited immune response in sepsis.

Effect of the GLP-1 analog exendin-4 and oxaliplatin on intrahepatic cholangiocarcinoma cell line and mouse model.

The influence of Glucagon-like peptide-1 (GLP-1) and Exendin-4 on development of intrahepatic cholangiocarcinoma (ICC) is evaluated in the study. In vitro tests, including acute toxicity test, cell colony formation assays, cells proliferation and apoptosis, transwell assay, were performed. An ICC in situ tumor animal model was established. Then, animals were randomly divided into four groups (n = 6): control, Exendin-4 treatment, oxaliplatin treatment and Exendin-4-oxaliplatin treatment. Animals in the Exendin-4 treatment and Exendin-4-oxaliplatin treatment groups received a subcutaneous injection of Exendin-4 (100 µg/kg/day) for 1 week, and then received oxaliplatin (10 mg/kg/week) by tail vein injection. Animals in the control group received PBS. Immunohistochemistry tests were used for PCNA, Ki67, Caspase 3 expression in tumor tissue. Results show that that, after incubation of human cholangiocarcinoma cell lines, HuCCTI and GLP-1, or HuCCTI and Exendin-4, colony formation number was sharply decreased. However, GLP-1, HuCCTI or Exendin-4 did not affect the colony of normal cells. Combination treatment with oxaliplatin and Exendin-4 can significantly inhibit tumor cells' proliferation and promote apoptosis. The combined effect is stronger than that of oxaliplatin or Exendin-4. Combination treatment with oxaliplatin and Exendin4 can significantly decrease Ki67 and PCNA proteins' expression in subcutaneous tumors of nude mice. The inhibitory effect of Combination treatment with oxaliplatin and Exendin4 is clearly stronger than that of oxaliplatin. In addition, Combination treatment with oxaliplatin and Exendin4 can significantly increase Caspase3 protein positive expression. In short, these results show that combination treatment with oxaliplatin and Exendin4 can inhibit tumor cells' proliferation, and promote apoptosis.

[Effects of resveratrol on matrix metalloproteinase-9 expression in hepatoma cells].

OBJECTIVE: To observe the effects of resveratrol on proliferation of human hepatocellular carcinoma cell line SMMC-7721 cells and expression of matrix metalloproteinase-9 (MMP-9) in vitro. METHODS: SMMC-7721 cells were treated with different concentrations of resveratrol for 24, 48 and 72 h, respectively. The effect of resveratrol on proliferation of SMMC-7721 cells was assessed with methyl thiazolyl tetrazolium (MTT). The expression of MMP-9 mRNA was determined by reverse transcription polymerase chain reaction (RT-PCR). MMP-9 protein was identified by Western blot analysis. RESULTS: Resveratrol could inhibit the proliferation of SMMC-7721 cells with dose- and time-dependent effects. Moreover, both MMP-9 mRNA expression and MMP-9 protein production were markedly reduced after resveratrol treatment. CONCLUSION: Resveratrol can inhibit the proliferation of SMMC-7721 cells and down-regulate MMP-9 expression. It is presumed that resveratrol may suppress the invasion and metastasis of hepatocellular carcinoma.

[Expression of heparanase and nuclear factor kappa B in pancreatic adenocarcinoma].

OBJECTIVE: To detect the expressions of heparanase and nuclear factor kappa B p65 (NF-kappaB p65) in pancreatic adenocarcinomas and analyze their relation to patients' prognosis and the regulatory mechanism of NF-kappaB on heparanase expression. METHODS: Heparanase and NF-kappaB p65 proteins in the tumor and adjacent tissues were detected by immunohistochemistry in 48 patients with pancreatic adenocarcinoma and analyzed for their clinicopathological significance. RESULTS: Heparanase and NF-kappaB p65 proteins were found in 30 (62.5%) and 22 (45.9%) tumor specimens, respectively, a rate significantly higher than that in the adjacent tissues. High heparanase expression was closely related to advanced TNM stage (P=0.031), lymph node metastasis (P=0.003) and decreased 3-year postoperative survival (20.0% vs 0%, P=0.001). NF-kappaB p65 expression was associated with lymph node metastasis (P=0.017) and distant metastasis (P=0.031), but had a higher positive rate in heparanase-positive cases than in heparanase-negative cases (P=0.018). Multivariate analysis showed that neither heparanase nor NF-kappaB p65 was the independent prognostic factors. CONCLUSION: Heparanase is overexpressed in pancreatic adenocarcinomas in association with decreased postoperative survival. NF-kappaB may up-regulate heparanase expression and promote heparanase-dependent tumor invasion and metastasis.

[Efficacy of modified acupotome combined with blocking therapy in patients with carpal tunnel syndrome].

OBJECTIVE: To observe the efficacy of modified acupotome combined with blocking therapy in patients with carpal tunnel syndrome (CTS). METHODS: Fifty-five patients with CTS were divided into three groups, which were modified acupotome group including 26 CTS patients with 28 lesions treated by modified acupotome combined with blocking therapy, traditional acupotome group including 14 CTS patients with 16 lesions treated by traditional acupotome combined with blocking therapy, and blocking therapy group including 15 CTS patients with 15 lesions only treated by local blocking. The treatment outcome and one-year recurrence rate were observed. RESULTS: The response rate and one-year recurrence rate after operation in the modified acupotome group were 85.7% (24/28) and 20.8% (5/24) respectively, which had no significant differences as compared with 81.3% (13/16) and 38.5% (5/13) in the traditional acupotome group. The response rate and one-year recurrence rate after operation in the above two groups were both improved significantly as compared with those in the blocking therapy group which were 46.7% (7/15) and 85.7% (6/7) respectively. There were no acupotome-related adverse effects and injuries observed in the modified acupotome group. CONCLUSION: The modified acupotome is a considerable treatment method for CTS with respect to its simple manipulation and high effectiveness.

[Efficacy of activated charcoal-epirubicin suspension for treatment of breast cancer with axillary metastasis].

OBJECTIVE: To investigate the effect of activated charcoal-epirubicin suspension (Epi-CH) for treatment of breast cancer and clearance of axillary lymph node metastasis. METHODS: Sixty patients with breast cancer of stages II-III were randomized into Epi-CH group (n=40) receiving injection with 10 mg Epi-CH in the tissue around the primary tumor 72 h before modified radical resection and control group (n=20) with 10 mg of aqueous epirubicin injection in the same region. The dissected axillary lymph nodes and the staining lymph nodes were counted. The concentration of epirubicin in the lymph nodes was detected by high-performance liquid chorography, and the specimens of lymph nodes were observed microscopically. RESULTS: In comparison with the control group, Epi-CH injection allowed dissection of 4.04 more lymph nodes (P<0.01) and resulted in the staining rate of the axillary lymph nodes of 86.9% (565/650). The proportion of the staining lymph nodes with diameter>1.0 cm was significantly lower than that with diameter 0.05). In Epi-CH group, the concentration of epirubicin in the stained lymph nodes was significantly higher than that in non-stained ones (443.0-/+123.1 vs 31.8-/+11.9 ng/g, P<0.01). Remarkable degeneration and necrosis could be observed microscopically in the stained lymph nodes. CONCLUSION: Epi-CH allows more effective treatment of breast cancer and clearance of axiliary metastasis by increasing the dissected lymph nodes and raising the concentration of epirubicin in the axillary lymph nodes.

[Impact of preoperative lymphatic chemotherapy on relapse and metastasis of breast cancer and its mechanism].

BACKGROUND & OBJECTIVE: Although it is reported that lymphatic chemotherapy could raise drug concentrations in local lymph nodes and prolong survival time of patients with gastrointestinal tumors, its effect on breast cancer has not been explored. This study was to explore the impact of lymphatic chemotherapy on relapse and metastasis of breast cancer, and to investigate the mechanism. METHODS: Sixty patients with breast cancer of stage II-III were randomized into 2 groups: 40 patients in Epi-CH (carbon activated absorbing epirubicin) group were injected with 10 mg of Epi-CH in the tissue around primary tumor 72 h before modified radical resection; 20 patients in control group were injected with 10 mg of aqueous epirubicin in the same region. The stained nodes full of tumor cells in Epi-CH group and non-stained nodes in control group were selected. The apoptotic index (AI) of cancer cells in metastatic axillary lymph node was calculated by TUNEL method; the expression of Fas/Fas-L proteins was examined by SP immunohistochemistry; the relapse and metastatic rate was compared. RESULTS: The AI of cancer cells in metastatic axillary lymph node was significantly higher in Epi-CH group than in control group [(9.5+/-2.7)% vs. (3.8+/-1.4)%, P<0.01]. The expression of Fas protein was significantly higher in Epi-CH group than in control group (P<0.05), but the expression of Fas-L protein had no difference between the 2 groups (P>0.05). No chemotherapy-related local and whole body reaction occurred in both groups. The relapse and metastatic rate was significantly lower in Epi-CH group than in control group (P<0.05). CONCLUSION: Preoperative Epi-CH lymphatic chemotherapy could suppress relapse and metastasis of breast cancer, which might through up-regulating expression of Fas protein and inducing apoptosis of axillary metastasis cells.

[Effect of preoperative lymphatic chemotherapy on Bcl-2 and Bax expression in axillary metastasis of breast cancer].

OBJECTIVE: To observe the expression of Bcl-2 and Bax proteins and cell apoptosis induced by preoperative lymphatic chemotherapy with epirubicin-activated carbon suspension (Epi-CH) in the cells of axillary metastatic lymph node of breast cancer and investigate the mechanism. METHODS: Sixty patients with breast cancer of stages II-III were randomly divided into two groups. Forty patients in Epi-CH group were injected with 10 mg Epi-CH in the tissue around the primary tumor or biopsy excision 72 h before operation. Twenty patients in the control group were injected with 10 mg epirubicin solution in the same region. The stained lymph nodes full of tumor cells in Epi-CH group and the non-stained nodes in the control group were selected for apoptotic detection by TUNEL method. The expression of Bcl-2 and Bax proteins were examined by SP immunohistochemistry. RESULTS: The apoptotic index of the metastatic cancer cells in Epi-CH group was increased remarkably in comparison with that in the control group [(9.5+/-2.7) % vs (3.8+/-1.4) %, P<0.01). Compared with the control group, the expression of Bcl-2 and Bax proteins were up-regulated significantly in Epi-CH group (P<0.05 and P<0.01, respectively), resulting in decreased ratio of Bcl-2/Bax. CONCLUSION: Lymphatic chemotherapy can promote cell apotosis in axillary metastasis of breast cancer, which may result from decreased ratio of Bcl-2/Bax.