Objective To establish and evaluate a diagnostic technique based on the AllGlo~(TM) probe for the invasive aspergillosis. Methods With the self-designed AllGlo~(TM) probes and primers and the standards, two standard curves of the real-time PCR based on AllGlo~(TM) probes were established respectively for A. flaws and A. fumigatus,then its specificity, sensitivity and reproducibility were evaluated respectively. Results The findings indicated that the standard curve of A. flavus was Y = - 3. 003X + 36. 825, and A. fumigatus' was Y = - 3. 052X + 38.016, and their interassay coefficient of variation respectively were 15.60% and 12. 94% , suggesting a good reproducibility. The lowest spore concentration they could be detected was 10 CFU/ml, which equated to 100-1000 copies of internal transcribed spacer (ITS)2 genes, suggesting a good sensibility. They didn't have cross-positive reaction with other fungus, human genome and bacteria, which indicated a good specificity. Conclusion The diagnostic technique based on the AllGlo?probe for the invasive aspergillosis possessed a good sensitivity, good specificity and deadly accuracy.

Objective To observe the effects of triptolide(TPL) on the anti-oncogene-APC gene of acute lymphoblastic leukemia cell line Jurkat in vitro. Methods The effects of TPL on proliferation Jurkat cells were assayed by using cell culture, MTT. The effects of TPL on APC gene of Jurkat cells were analyzed by nested methylation specific PCR and RT-PCR. The effects of TPL on the proteinum expression of APC gene were detected by Western blotting analysis. Results Following the treatment of TPL, the cell proliferation rate was degraded as the treatment concentration increased and the culture time extended. The effects were dose and time-dependent. The 48 hour IC50 was 19.7 ng/ml. TPL can reverse hypermethylation of APC gene,and induce the expression of the mRNA and the proteinum. Conclusion Low dose TPL could depress the proliferation rate of Jurkat. The possible mechanism might be its reversing the hypermethylation of APC gene and activiting the expression of APC gene.

Objective To investigate the expression of interleukin-35 (IL-35) in patients with sepsis and multiple organ dysfunction syndrome (MODS),and to explore its effect on disease prognosis.Methods A prospective study was conducted.Twenty-two patients with sepsis and MODS were selected as study group,and 20 healthy volunteers were selected as controls.Blood samples of the patients and the volunteers were taken within 1 hour of the patient's visit,and cytokines,such as IL-35,IL-4,IL-10,IL-17,INF-γ,and TGF-β,were simultaneously detected by enzyme-linked immunosorbent assay (ELISA).The expression level of CD4+CD25+FOXP3+ Treg cells was detected by flow cytometry,and the acute physiology and chronic health status scoring system Ⅱ (APACHE Ⅱ) was calculated.According to the survival outcome at 28 days after admission,the expression levels of IL-35 in the survival group (12 cases)and the death group (8 cases) were compared.The Spearman method was used to analyze the correlation between IL-35 level and the.above indicators in patients with sepsis and MODS.Results Compared with the control group,the IL-35 levels were significantly increased in the study group (P＜0.05),and the IL-35 levels began to increase gradually on the first day of the disease,and significantly higher on the 7th day (P＜0.01).The number of CD4+CD25+FOXP3+regulatory T cells increased in the study group,especially on the 4th day (P＜0.01),and on the first day of the disease,the IL-35 levels were positively correlated to the number of CD4+CD25+FOXP3+Treg cells (r=0.60,P＜0.05).Compared with the control group,the levels of IL-10,IL-4,IL-17,INF-γ and TGF-β in the study group increased gradually with the course of disease,and were significantly higher on the 7th day than those on the 1st day and the 4th day (P＜0.05).The IL-35 levels in the study group were positively correlated with INF-γ and TGF-β cytokines (P＜0.05),whereas there was no significant relationship between the levels of IL-10,IL-4,and IL-17 (P＞0.05).The IL-35 level in the death group was significantly lower than that in the survival group (P＜0.05).IL-35 levels in the survival and death groups were analyzed by ROC curve with a AUC of 0.78 (P=0.03).The ~ IL-35 level was negatively correlated with the APACHE-Ⅱ score in the study group (r=-0.78,P＜0.01).Conclusions The IL-35 levels in patients with sepsis and MODS is significantly higher than that in healthy controls,and negatively correlated with APACHE Ⅱ score.The level of IL-35 has an important implication for the prognosis for patients with sepsis.