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Objective To explore the early laboratory indicators for risk of intravenous immunoglobin-resistant Kawasaki diseases. Methods The clinical data were retrospectively analyzed in 881 Kawasaki disease patients (group A: 26 cases of intravenous immunoglobin-resistant; group B: 855 cases of intravenous immunoglobin-sensitive) from July 1, 2015 to June 30, 2016. After 1:3 matching with age and sex, the regression model for each of variables including sex, age, fever days, temperature, red blood cell count (RBC), white blood cell count (WBC), neutrophil (N), lymphocyte (L), platelet count (PLT) and C reactive protein (CRP), was constructed by conditional logistic regression analysis. Results Compared with group B, group A had significantly lower RBC count and higher PLT (P<0.05). Logistic regression analysis showed that, with the age, the regression model was Y=-2.87+0.01×PLT (PLT OR=1.01, 95％ CI: 1.00～1.01, P<0.01); with the sex, Y=-32.98+0.44×WBC+0.28× N+0.01×PLT (WBC OR=1.55, 95％ CI: 1.17～2.05, P<0.01; N％ OR=1.32, 95％ CI: 1.04～1.68, P<0.05; PLT OR=1.01, 95％ CI 1.00～1.01, P<0.01). Conclusion In case that abnormally high levels of PLT exist in confirmed Kawasaki disease, it should be aware of possibility of the intravenous immunoglobin-resistant Kawasaki disease.
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AIM: To investigate the effects of TNF receptor-associated death domains (TRADD) of latent membrane protein 1 (LMP1) on the proliferation of nasopharyngeal cancer SP18 cells.METHODS: The SP18-LMP1 cells and SP18-LMP1TRADD cells, which expressed LMP1 and LMP1 TRADD proteins, respectively, were established.The proliferation of SP18 cells affected by LMP1TRADD was detected by cell counting to analyze the cell growth curve, and by colony formation assay, soft agar formation assay, and flow cytometry.Moreover, the expression profile of differential genes between SP18-LMP1 cells and SP18-LMP1TRADD cells was analyzed by gene chips.RESULTS: The cell growth curve, and the results of colony formation and soft agar formation displayed that the growth velocity and colony forming ability of SP18-LMP1 cells were stronger than those of SP18-LMP1TRADD cells (P<0.01).The results of flow cytometry analysis showed that the proliferation index of SP18-LMP1 cells was higher than that of SP18-LMP1TRADD cells (P<0.01).Sixty-three differentially expressed genes associated with cell proliferation were screened out, in which 33 genes were up-regulated and 30 were down-regulated in the SP18-LMP1TRADD cells.CONCLUSION: TRADD active region is an important functional site of LMP1 to promote the proliferation of SP18 cells.LMP1 may improve the cell proliferation index and induce the proliferation of SP18 cells through TRADD.
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Objective: This study investigates the biological effects and explores the molecular mechanisms of epigallocate-chin-3-gallate (EGCG) on the apoptosis of the human gastric cancer MGC-803 cells. Methods: After treatment with EGCG, cell apopto-sis was verified by flow cytometry with Annexin V and propidium iodide staining, DNA agarose gel electrophoresis, and transmission electron microscopy. The expression profiles of the apoptosis-related genes in the MGC-803 cells with or without treatment by EGCG for 12 h (100 μmol/L), was identified using SuperArray Human Apoptosis Gene Array. The upregulated Fas-L gene and down-regulated Bag-1 gene were confirmed by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Results: When the MGC-803 cells were treated with EGCG at 25, 50, 100, and 200 μmol/L for 24 h, evident sub-diploid peaks were observed. Under treat-ment with 100 μmol/L for 4, 8, 12, and 24 h, the number of early apoptotic cells was greatly increased. When the cells were treated with 100 μmol/L for 24 h, the DNA extracted from the cells displayed a characteristic ladder pattern with agarose gel electrophoresis. Typi-cal morphological changes were observed by electron microscopy, including cell shrinkage, karyo-pyknosis, and the formation of apop-totic bodies. The differential expressions of eight apoptosis-associated genes were determined by gene array detection. The results of Fas-L and Bag-1 selected for RT-PCR and Western blot were consistent with those of gene array. Conclusion: EGCG induces apoptosis in MGC-803 cells, which might be mediated by a number of specific genes and various signal transduction pathways.
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Background and purpose:To investigate the induction of apoptosis by epigallocatechin-3-gallate(EGCG) in xenograft nude mice with human gastric cancer cells and its molecular mechanism. Methods:Human gastric cancer cells were planted into nude mice in order to establish the cancer model, the different dosages of EGCG were injected intraperitoneally in the nude mice. After treatment, flow cytometry (FCM) was used to detect the apoptosis of implanted tumor cells. Immunohistochemical staining was used to detect the expression of apoptosis-related genes like Bal-2 and Bax in implanted tumor.Results:EGCG significantly inhibited tumor growth after being injecting intraperitoneally in the nude mice. The apoptotic cells in implanted tumor could be detected by flow cytometry with PI staining. The expressions of Bax、Caspase-3 were upregulated and Bcl-2 expression was downregulated in implanted tumor.Conclusions:EGCG could significantly inhibit tumor growth in xenograft nude mice with human gastric cancer cells through inducing apoptosis. The down-regulation of Bcl-2 expression and up-regulation of Bax expression observed could result in the activation of Caspase-3, the pathway might account for the induction of apoptosis.
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Aim The aim of this work was to study the effect of diallyl disulfide (DADS) on human Leukemia cell line HL 60, and investgate the mechanisms of its antitumor effect. Methods HL 60 cells growth inhibition was measured by MTT assay. Cell apoptosis was inspected by flow cytometry,TUNEL assay and acridine orange fluorescent staining methods. The protein levels of Bcl 2, Bax were determined using immunohistochemical technique. Results MTT assay showed that DADS significantly inhibited the growth of HL 60 cells. After 24 hours of exposure to DADS, Partial cells presented characteristic morphological changes of apoptosis under the electron microscope, including cell shrinkage, nuclear condensation, and formation of apoptotic bodies. Some typical subdiploid peaks before G 0/G 1 phase were observed. Flow cytometry analysis revealed that the apoptosis rates were increased and the TUNEL assay showed the apoptosis index increased in occordance with increase of concentration of DADS. SP immunohistochemistry revealed that the Bax expression was increased while Bcl 2 expressed was decreased. Conclusion Diallyl disulfide could significantly induce apoptosis of human Leukemia cell HL 60, Apoptosis of tumor cells is closely associated with down regulation of the ratio of bcl 2/bax.
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Objective To investigate the effect of Rb protein expression on the clinical response of the colorectal cancer patients perfused with 5-Fu before operation. Methods The expression of Rb protein in 34 cases of the colorectal cancer sample was determined by streptavidin-peroxidase(S-P) immunohistochemical method and the positive reactions were analysed by ICM-100 Cell DNA Picture Analysis System. The apoptotic rate of colorectal cancer cells was examined by the TUNEL staining. The chemosensistivity of the colorectal cancer cells were tested by the MTT colorimetric assay of agar culture. The clinical response of the colorectal cancer patients perfused with 5-Fu before operation was observed simultaneously. Results ⑴There was the negative correlation between the resistance index of the colorectal cancer cells to 5-Fu (IC50/10-1 peak concentration) and the apoptotic rate of the colorectal cancer cells induced by 5-Fu (?=-0.53,t=-4.91,P1.0 belongs to the group of resistance,
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Objective To explore the value of morphometric quantitative parameters in differential diagnosis of common thyroid benign and malignant tumors, and establish a data bank as a basis for diagnosing diseases with the help of computer. Methods Using PIPS-2020 image analysis system to make a morphometric analysis in 32 cases of thyroid benign papillary lesion (TBPL), 32 cases of thyroid papillary adenocarcinoma(TPAC), 27 cases of thyroid follicular adenoma(TFA) and 24 cases of thyroid follicular adenocarcinoma(TFAC). Results Among many morphometric quantitative parameters of nuclei, eight including area, circumferential length, diameter,volume,long diameter, short diameter, largest to smallest diameter ratio, and circular rate were selected to analyze. The eight morphometric quantitative parameters between TFA and TFAC had a highly significant difference(P0.01). Conclusion These nuclear parameters have some value in differentiation of common thyroid benign and malignant tumors.
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Objective To investigate the influences of mutant p53 protein expression on the resistance of breast cancer cells to adriamycin and efficiency of adriamycin chemotherapy of the pre-operative patients with breast carcinoma.Methods Mutant p53 protein expression in 9 cases of breast cancer was examined by using inmunohistochemical method and image semiquantified analysis.The response of human breast cancer cells to adriamycin was tested with MTT assay in order to judge the effects of mutant p53 protein expression on the pre-operative pateints with breast carcinoma treated with adriamycin chemotherapy.Results ⑴Mutant p53 protein expression in 7 cases of the human breast cancer samples were positive,the positive rate was 77%(7/9).⑵There was positive relationship between expression degree of mutant p53 protein(positive relative area) and resistance index(IC50:1/10 peak concentration) of human breast cancer cells to adriamycin(r=0 7956,P
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Background and purpose:Anticancer mechanism of epigallocatechin-3-gallate(EGCG)remains unclear.This study investigated the role of reactive oxygen species(ROS)in epigallocatechin-3-gallate(EGCG)-induced apoptosis in human gastric cancer MGC803 cells.Methods:The inhibition of MGC803 cells growth was measured by MTT assay.Apoptosis of MGC803 cells was studied by using the AO/EB fluorescence stain.Flow cytometry was used to detect the intracellular ROS level and the rate of apoptosis.Results:EGCG could induce apoptosis of MGC803 cells and increased in the intracellular ROS level.However,after treatment with N-acetyl-L-cystein and an athiol-containing antioxidant,the inhibitory effect of EGCG on MGC803 cells was significantly weakened.The apoptotic rate of the cells and the activity of the intracellular ROS level also decreased dramatically.Conclusion:EGCG can induce apoptosis of MGC803 cells.In turn,the ROS inhibitor can significantly inhibit the apoptosis induced by EGCG in MGC803 cells.These results suggest that the cellular generation of ROS plays a role in initiating EGCG-mediated apoptosis of MGC803 cells.