Objective To discuss the anti-inflammatory effect and its possible mechanism of different insulin concentration on IκB, NF-κB and TNF-α mRNA in monocytes stimulated by lipopolysaccharide and to detect the key molecule P-Akt in PI3K/ Akt signaling pathway at the cellular level. Methods The peripheral blood (10 ml)is collected from 30 type 2 diabetics (T2DM). Then, the mononuclear cells are centrifugally separated based on density gradient and divided into five different groups:the control (Con)group; the low concentration insulin 100 mU/L(L-Ins)group; the combination of low concentration insulin 100 mU/L and LY-294002 10 μmol/L(L-Ins+LY) group; the high concentration insulin 1000 mU/L (H-Ins) group, The combination of high concentration insulin 1000 mU/L and LY- 294002 10 μmol/L(H-Ins+LY) group. Two subgroups were set in each category. After incubating for 24 hours, the lipopolysaccharide (100 ng/ml)was added and the mononuclear cell culture of peripheral blood was continued. One hour later, the activities of P-Akt, IκBα and NF-κB of one subgroup were detected using western blot; two hours later, monocytes of the other subgroup were collected and the level of TNF-a mRNA was detected using real-time PCR Results Compared with Con group, the expressions of P-Akt and IkB were higher in L-Ins group and H-Ins group (P<0. 05), the expressions of NF-κB and TNF-a mRNA were lower in L-Ins group and H-Ins group (P<0. 05). Compared with L-Ins group, the expressions of P-Akt and IκB were higher in H-Ins group (P<0. 05), but the expressions of NF-κB and TNF-a mRNA were lower (P< 0. 05). Compared with L-Ins + LY group (H-Ins + LY group), the expressions of IκB and P-Akt were higher(P<0. 05)and the expressions of NF-κB and TNF-a mRNA were lower in L-Ins group (H-Ins group)(P<0. 05). Compared with Con group, no significant variations were shown in the expressions of P-Akt, IkB and NF-κB in L-Ins+LY group and H-Ins+LY group (P>0. 05) except for the high expressions of TNF-a mRNA (P<0. 05). There is no significant difference between L-Ins+LY group and H-Ins+LY group (P>0. 05). Conclusion The direct anti-inflammatory effect of insulin is verified in a dose dependent manner. Insulin may regulate the synthesis and secretion of inflammatory cytokines by activating PI3K/Akt pathway, increasing IkB and affecting the state of NF-κB. Insulin may increase the synthesis and secretion of inflammatory cytokines through other pathways when the PI3K/Akt pathway is blocked.

Purpose To investigate the consistency of the"Standards and guidelines for the interpretation and reporting of sequence variants in cancer"published in 2017 by the Associa-tion for Molecular Pathology(AMP)/American Society of Clini-cal Oncology(ASCO)/College of American Pathologists(CAP).Methods Sixty variants of 26 genes from 11 types of cancer were selected.5 professionals from four hospitals e-quipped with in-hospital NGS detection ability were used to in-terpret the treatment,diagnosis and progenosis respectivly.In the first phase of the study,each researcher rated the variants individually according to their own understanding of the 2017 guideline.In the second phase,the details of the guidelines'e-valuation principles were discussed and interpreted again after reaching a consensus.Results Eleven principles recognized by all participants were summarized as a supplement to interpreta-tion.Fleiss consistency showed that the consistency of interpre-tation of treatment and prognostic significance in the second stage was higher than that in the first phase(κ value was 0.166 vs 0.276,0.014 vs 0.185).The consistency of interpretation of diagnostic significance in the second stage was lower than that in the first phase(κ value was 0.454 vs 0.035).Conclusion There is inconsistency in the clinical interpretation of tumor gene variation among different laboratories.It is feasible for laborato-ries to establish a mutually recognized interpretation system for the clinical diagnosis,treatment,and prognosis of tumors.