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Objective:To investigate the radiosensitizing effect and underlying mechanism of STING agonist (c-di-AMP) on cutaneous melanoma cells.Methods:Human cutaneous melanoma cells (A375) were divided into four groups: the control group, 10 μmol/L c-di-AMP group, X-ray irradiation group and X-ray irradiation combined with c-di-AMP group. The radiosensitizing effect of c-di-AMP on A375 cells was detected by CCK-8-based viability assay, lactate dehydrogenase (LDH) release assay, flow cytometry-based apoptosis assay, and colony formation assay. Western blot analysis was used to determine the expressions of cell death-related proteins.Results:In combination with 10 Gy X-ray irradiation, 10 μmol/L c-di-AMP showed significant radiosensitization effect in A375 cells, which was evidenced by decreased cell activity ( t=5.11, P<0.05), increased cytotoxicity ( t=10.15, P<0.05) and cell apoptosis ( t=4.41, P<0.05) and reduced clone viability( t=6.30, 3.55, 5.45, 3.55, P<0.05). The calculated radiosensitization ratio of c-di-AMP to A375 cells was 1.88. Moreover, 10 μmol/L c-di-AMP further increased the expressions of cell death-related proteins induced by radiation in A375 cells. Conclusions:The STING agonist c-di-AMP can be used as a radiosensitizer for cutaneous melanoma, which may provide a novel strategy for radiotherapy.
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Background: C-type lectin domain family 4 member D (CLEC4D) can induce Th17 cell differentiation and regulate IL-17A expression in systemic fungal infection, and whether CLEC4D plays a role in intestinal immunity has not been reported at home and abroad. Aims: To investigate the expressions and clinical significance of CLEC4D and CARD9 in colon tissue in patients with inflammatory bowel disease (IBD). Methods: A total of 48 IBD patients from October 2016 to June 2018 at the Second Affiliated Hospital of Zhengzhou University were enrolled, including 36 patients with ulcerative colitis (UC) and 12 patients with Crohn's disease (CD). And 30 adjacent normal colon tissues in patients with colon cancer were served as controls. Immunohistochemistry was used to detect the expressions of CLEC4D and CARD9 in colon tissue. And their correlations with clinical characteristics were analyzed. Results: The expressions of CLEC4D and CARD9 in UC group and CD group were significantly higher than those in control group (P0.05). The expressions of CLEC4D and CARD9 in UC group were positively correlated with Mayo score and Baron grade (P0.05). The expressions of CLEC4D and CARD9 in IBD patients were positively correlated with CRP (P0.05). Conclusions: The expressions of CLEC4D and CARD9 are elevated in patients with IBD and are closely related to patients' condition. CLEC4D may participate in the intestinal immunity of IBD through the CARD9-related pathway.
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Inflammatory bowel disease (IBD)is an autoimmune disease leading to diarrhea,abdominal pain,and weight loss. The pathogenesis of diarrhea remains unclear,however,increasing evidence has shown that the epithelial sodium channel (ENaC)is associated with diarrhea. ENaC is crucial in the control of sodium homeostasis,extracellular fluid volume,blood pressure. This article reviewed advances in study on relationship between ENaC and IBD.
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Objective To evaluate the relationship between sternum protection and bone marrow suppression in postoperative radiotherapy for esophageal carcinoma. Methods Total of 98 postoperative patients with thoracic esophageal carcinoma were randomly divided into experimental group (52 cases) and control group (46 cases). All patients were given intensity-modulated radiotherapy (IMRT), with the dose of 50-50.4 Gy. The patients in experimental group were irradiated by 6 fields (4-fields in front, 2-fields behind) which were crossed to avoid direct exposure to the sternum. The patients in control group were irradiated by 5 fields (3-fields in front, 2-fields behind) with front-middle of the field passing through the sternum. Concurrently all patients received 2 cycles of cisplatin chemotherapy. Results Dmean, V20 and V30 of the sternum in the experimental group were (20.21 ±3.60) Gy, (40.78 ±7.19) % and (33.78 ±9.44) %, which were lower than those in the control group [(30.91±5.21) Gy, (81.01±4.81) %, (51.60±6.84) %], respectively (P 0.05). Both the incidence rates of bone marrow suppression at 14th day and 35th day after radiotherapy were significantly higher in the control group (52.2%, 73.9%) than those in the experimental group (28.8 %, 50.0 %) (P< 0.05), and the incidence rate of bone marrow suppression at 7th day after radiotherapy was similar between the two groups. Conclusion Protecting and sketching for sternum in postoperative radiotherapy for esophageal carcinoma can reduce the incidence of bone marrow suppression effectively, which would not increase the radiation dose in the lung, heart and spinal cord.
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To obtain the expression of Isoleucine Zipper modified soluble CD40L (IZ-sCD40L) in Pichia pastoris, firstly, DNA fragment of IZ-sCD40L was obtained by PCR and over-lap PCR . Then the expression vector pPICZaA-IZ-sCD40L was constructed. Nucleotide sequencing analysis indicated that the DNA fragment of IZ-sCD40L was correctly inserted into the pPICZaA vector. Linearized pPICZ(alpha)A-IZ-sCD40L was introduced into Pichia pastoris GS115. Positive clone was selected by PCR and its phenotype was determined. The positive clone was introduced with methanol. The results of SDS-PAGE and Western blot showed that product was recombinant Isoleucine Zipper modified soluble CD40L fusion protein.
Subject(s)
Angiotensin II , CD40 Ligand , Genetics , Cloning, Molecular , Isoleucine , Genetics , Pichia , Genetics , Metabolism , Plasmids , Polymerase Chain Reaction , Methods , Recombinant Fusion Proteins , Genetics , Recombination, GeneticABSTRACT
OBJECTIVE:To observe the short-term curative effects of 3-dimensional conformal radiotherapy(3DCRT) combined with taxotere on locally advanced non-small cell lung cancer.METHODS:83 patients Ⅲ a or Ⅲ b with locally advanced non-small cell lung cancer were divided randomly into the treatment and control group.The treatment group were treated with 3DCRT in combination with taxotere,while the control group were treated with single 3DCRT.RESULTS:The effective rates in the treatment and control group were 88.37% and 62.50%(P