1 The Effect Of Licl On The Non-canonical WNT Signal Transduction Pathway In Leukemia Cell Lines 2 The Relationship Between Infection And The Etiology Of Kawasaki Disease

Objective: To investigate the expression of WNT receptors in the leukemia cells treated before and after Licl, and to explore the effect of Licl on the non-canonical WNT signal transduction pathway in Leukemia cell lines.Methods: 1 To construct the Licl inhibited cell lines by culturing the Jurkat, K562, HL-60 cells with different Licl concentration; Experimental groups: Jurkat, K562, HL-60 cells inhibited by Licl. Control groups: Jurkat, K562, HL-60 cells and ITP. 2 RT-PCR was used to assay the RNA expression of Frizzled-2, Frizzled-5, Frizzled-7, Frizzled-8, LRP-6 and ROR2 in experimental groups and control groups. 3 Western Blot was used to assay the protein expression of Frizzled-5 and ROR2 in the experimental groups and control groups. 4 To assay the cell cycle of the experimental groups and control groups by the methods of FCM. 5 To assay the cell proliferation of the experimental groups and control groups by the methods of MTT.Results: 1 The best inhibited Licl concentration was 40mmol/L, and the experimental groups were successfully constructed. 2 For the gene expression of Frizzled-2, Frizzled-7, Frizzled-8 and LRP-6, no differences were found between the experimental groups and control groups. There were no differences of the gene expression of ROR2 in the K562 and HL-60 cell lines treated before and after Licl. While for the expression of ROR2, there was significant difference in Jurkat cells treated before and after Licl. 3 For the protein expression of ROR2, no differences were found in K562 and HL-60 cell lines treated before and after Licl; While significant difference was found in Jurkat cell line treated before and after Licl.4 For the cell cycle, the G2 ratio of the experimental Jurkat cell was lower than that of the control Jurkat cell; while in K562 and HL-60 cell lines treated before and after Licl, no differences were found. 5 The proliferation of the experimental Jurkat cell line was lower than that of the control Jurkat cell; while no differences were found in K562 and HL-60 cell lines treated before and after Licl.Conclusions: The inhibition role of Licl on Jurkat cell line was dose dependent under the range of 40mmol/L. As the concentration raised up, the inhibition effect was enhanced; and the best inhibition concentration was 40mmol/L. While Licl played no inhibition roles on K5 62,HL-60 and ITP cells. 2 For the expression of Frizzled/LRP and ROR2, no differences were found in K562 and HL-60 cell lines treated before and after Licl; While for the expression of ROR2, there was significant difference in Jurkat cell line treated before and after Licl.3 For the cell cycle and proliferation, Licl plays a significant inhibition role on Jurkat cell line.We supposed that Licl could decrease the expression of ROR2, and inhibit the non-canonical WNT signal transduction pathway. While Licl played no inhibition role on the normal cell, so Licl could be a new way for the clinical treatment of ALL. Objection: To investigate the pathogen related to Kawasaki disease ( KD) and to explore the relationship between Kawasaki disease and infection.Methods:1 System evaluation:Search the articles related to the etiology of KD that published in the CNKI, pubmed, Embase, medline datebases from Jan 1979 to Oct 2010. Only Chinese and English language were included. Include the original articles, evaluate the quality and extract the information, than analysis the date by Revman 5 provided by the Cochrane.2 Clinical empirical study: There were three groups in this research: Kawasaki disease (KD), Acute infectious disease children (IC) and healthy children(HC). We assayed the levels of SPA, TSST-1, SE, ASP, HSPA-Ag, and HASP-Ag in the three groups by the methods of ELISA. For the KD group and IC group, bacterial cultures were obtained for the blood, throat swab and stool.3 Retrospective analysis: 266 KD children hospitalized during Jan, 2002 to Dec, 2008 were reviewed and analyzed from the next three parts: the results of hemoculture, the positive rate of hemoculture in KD patients of different age groups, and the relationship between hemoculture and CAL.Results:1 12 articles were included, containing 5 kinds of pathogens. 593 cases of KD children and 1154 cases of control children were studied. 1 Staphylococcus aureus and group A streptococcus: the odds ratio was 4.26, 95% confidence interval was [2.54,7.16], P<0.00001. 2 Mycoplasma and Chlamydia: the odds ratio was 3.41, 95% confidence interval was [1.99,5.84], P<0.00001. 3 Human mini virus B19:the odds ratio was 6.58, 95% confidence interval was [2.37,18.25], P=0.0003. 4 Human coronavirus NL-63: the odds ratio was 0.75, 95% confidence interval [0.20,2.78], P=0.66.2 The levels of SPA and TSST-1 in the sera of KD group were 1.93±0.35 and 6.50±1.15, and in the IC group were 1.19±0.24 and 6.78±1.03, while in the HC group were 1.27±0.29 and 5.42±0.92. There was significant difference among the three groups(P<0.05). For the levels of SE, ASP, HSPA-Ag and HASP-Ag, no differences were found among the three groups. The positive ratio of hemoculture in KD group was significantly higher than that in IC groups(P<0.05).3 There were statistic differences among the positive ratios of different age groups (p<0.05). The CAL ratio of positive hemoculture cases were higher than that of negative hemoculture cases(P<0.05). Conclusions:1 The System evaluation suggested that pathogen infection was related to the development of KD. Staphylococcus aureus, group A streptococcus, Human mini virus B19, Mycoplasma and Chlamydia were related to the development of Kawasaki disease, while Human coronavirus NL-63 had no relationship with KD. But as the studies included were few, the quality of those articles were low, further conclusions needs to be drawn through a number of well-designed, large sample of prospective studies.2 The superantigen reaction induced by SPA and TSST-1 may related to the development of KD, but we found no relationship between group A streptococcus superantigens and KD. The positive ratio of hemoculture in KD was higher than that in control groups.3 The positive ratio of hemoculture in elderly children was significantly higher than that in infantum. Maybe there was certain correlation between positive hemoculture and CAL.In a word, Bacterial infection may involved in the development of KD.The positive hemoculture may be a risk factor of CAL.