A Multicenter Study of Intravenous Immunoglobulin Non-response in Kawasaki Disease.

To investigate the relationship between the risk factors associated with intravenous immunoglobulin (IVIG) non-response and the incidence of coronary artery lesions (CAL) in patients with Kawasaki disease (KD). A retrospective study was performed on clinical records of 1953 KD patients who were admitted to hospitals in Shanghai, China, between 1998 and 2007. Related clinical and laboratory findings were studied using univariate and multivariate statistical analyses. Of the 1953 KD patients, 133 (6.8 %) were unresponsive to IVIG therapy, and 356 (18.6 %) developed CAL. The incidence of CAL in the non-responsive IVIG group was significantly different from that in the responsive IVIG group (31.3 vs. 17.6 %). The incidence of IVIG non-response was significantly lower in the patients who received sufficient doses of IVIG than in the patients who received insufficient doses (5.2 vs. 18.1 %). A logistic regression analysis of 1295 patients who received sufficient IVIG doses indicated that cervical lymph node enlargement, CAL, erythrocyte sedimentation rate (ESR) ≥75 mm/h, and platelet count (PLT) ≥530 × 10(9)/L were independent risk factors of IVIG non-response. IVIG non-responders are prone to develop CAL. Initiation of therapy with sufficient IVIG doses at the early stage of the disease is crucial for preventing IVIG non-response. Lymph node enlargement, ESR ≥75 mm/h, and PLT count ≥530 × 10(9)/L are independent risk factors for predicting non-response to sufficient IVIG doses. For patients with the tendency of being unresponsive to IVIG therapy, treatment using sufficient IVIG doses combined with hormones or immunosuppressive agents should be considered to reduce the incidence of IVIG non-response and CAL.

Interacting domains of P14-3-3 and actin involved in protein-protein interactions of living cells.

14-3-3 proteins are conserved regulatory proteins present in all eukaryotic cells that control numerous cellular activities via targeted protein interactions. To elucidate the interaction between P14-3-3 from Physarum polycephalum and actin in living cells, PCR and DNA recombination were used to generate various P14-3-3 and actin constructs. Yeast two-hybrid assay and FRET were employed to characterize the interaction between P14-3-3 and actin. The two-hybrid assay indicated that P14-3-3 N-terminal 76-108 amino acids and the C-terminal 207-216 amino acids played an important role in mediating interactions with actin, and the actin N-terminal 1-54 amino acids and the C-terminal 326-376 amino acids are also crucial in the interactions with the mPa, a P14-3-3 with mutations at Ser62 (Ser62 â†’ Gly62). Mutations to potential phosphorylation sites did not affect interactions between P14-3-3 and actin. FRET results demonstrated that P14-3-3 co-localized with actin with a FRET efficiency of 22.2% and a distance of 7.4 nm and that P14-3-3 N-terminal 76-108 and C-terminal 207-216 amino acids were important in mediating this interaction, the truncated actin peptides without either the N-terminal 1-54 or C-terminal 326-376 amino acids interacted with P14-3-3, consistent with the results obtained from the yeast two-hybrid assay. Based on data obtained, we identified critical actin and P14-3-3 contact regions.

TAGLN mediated stiffness-regulated ovarian cancer progression via RhoA/ROCK pathway.

BACKGROUND: Ovarian cancer (OC) progression is an unmet medical challenge. Since omental metastases were palpated harder than their primary counterparts during cytoreductive surgery of patients with epithelial ovarian cancer (EOC), we were inspired to investigate OC progression from the perspective of biomechanics. METHODS: Atomic Force Microscope (AFM) was used to measure the Young's modulus of tissues. The collagen-coated polyacrylamide hydrogel (PA gel) system was prepared to mimic the soft and stiff substrates in vitro. The effect of TAGLN was evaluated both in vitro and in vivo using transwell assay, immunofluorescence, western blot analysis and immunohistochemistry. RESULTS: We quantitatively confirmed that omental metastases were stiffer and more abundant in desmoplasia compared with paired primary tumors, and further demonstrated that matrix stiffness could notably regulate OC progression. Remarkably, TAGLN, encoding an actin cross-linking/gelling protein, was identified as a potent mechanosensitive gene that could form a regulation loop with Src activation reacting to environmental stiffness, thus mediating stiffness-regulated OC progression through regulating RhoA/ROCK pathway. CONCLUSIONS: These data demonstrate that targeting extra-cellular matrix (ECM) stiffness could probably hamper OC progression, and of note, targeting TAGLN might provide promising clinical therapeutic value for OC therapy.

mTOR inhibition overcomes primary and acquired resistance to Wee1 inhibition by augmenting replication stress in epithelial ovarian cancers.

Epithelial ovarian cancer is characterized by universal TP53 mutations, which result in G1/S checkpoint deficiencies. Therefore, it is hypothesized that the abrogation of the G2/M checkpoint with Wee1 inhibitor might preferentially sensitize TP53-defective ovarian cancer cells. Given the extremely high molecular diversity in ovarian cancer, one approach to improving the clinical efficacy is to identify drug combinations that either broaden the applicable spectrum or circumvent resistance. Here, through a high-throughput unbiased proteomic profiling (RPPA), we found the complementary activated mTOR pathway contributes greatly to Wee1 inhibitor resistance. A combination of Wee1 and mTOR inhibits synergistically inhibiting tumor growth in ovarian cancer cell lines and patient-derived xenograft that closely mimic the heterogeneity of patient tumors. Mechanistically, dual Wee1/mTOR inhibition induced massive DNA replication stress, leading to fork stalling and DNA damage. Moreover, we found that the addition of nucleotide metabolic substrate dNTPs alleviated replication stress, restored the cell cycle and reduced apoptosis to some extent, supporting dNTPs depletion is necessary for the synergy between Wee1 and mTOR inhibits. These results suggest that our study opening up a wider therapeutic window of Wee1 inhibitor for the treatment in epithelial ovarian cancers.