[ANXA8 Regulates Proliferation of Human Non-Small Lung Cancer Cells A549 via EGFR-AKT-mTOR Signaling Pathway].

Annexin A8 (ANXA8) is a member of the annexin family, which had been reported to regulate multiple cancer cellular processes including proliferation, metastasis and inflammation. However, the specific role of ANXA8 in lung cancer cell biology remains unknown. Our previous transcriptome study revealed that ANXA8 mRNA was downregulated in curcumin analog (MHMD) -treated human non-small lung cancer cells (A549 cell line). Here, we continued to study the ANXA8 expression in A549 cells using reverse transcription-quantitative PCR and Western blotting, compared with that in human normal bronchial epithelium cells (BE-AS-2B cell line). Overexpression of ANXA8 via transfection of pEGFP-ANXA8 recombinant vector contributed to the proliferation and migration of A549 cells. Moreover, the cell cycle protein cyclin E1 was upregulated in ANXA8-transfected A549 cells. Knockdown of ANXA8 using an RNA interference technique decreased A549 cell viability and restrained their migration in vitro. The expression levels of multiple cellular factors, including EGFR, PI3K, Akt, mTOR, p70S6K and 4EBP1, in the epidermal growth factor receptor (EGFR) signaling pathway were also altered by ANXA8 knockdown or overexpression in A549 cells, which confirmed the activation of the EGFR/Akt/mTOR signaling pathway by ANXA8. The present results provided evidence to support further investigation of the functional identification of ANXA8 in lung cancer cells in the future.

[Study on the risk factors for hemorrhagic transformation of cerebellar infarction after posterior fossa decompression surgery].

Objective: To investigate the related risk factors for hemorrhagic transformation (HT) of cerebellar infarction after posterior fossa decompression surgery. Methods: A total of 91 patients with cerebellar infarction were treated by posterior fossa decompression surgery in Department of Neurosurgery of Jinhua Municipal Central Hospital from Jan 2010 to Jan 2018, were selected as study subjects. The HT group included 17 cases, while the Non-HT group included 74 cases. The clinical data of the two groups were analyzed retrospectively, the univariate and non-conditional lgistic regression analysis were performed to detect the relevant risk factors for hemorrhagic transformation of cerebellar infarction after posterior fossa decompression surgery. Results: By univariate analysis, the differences of these seven risk factors, the large area cerebellar infarction (the diameter of area was larger than 5 cm), pre-op thrombolysis, pre-op mild HT, oral anticoagulants, atrial fibrillation, hyperglycemia and fluctuation of BP in post-op, between two groups were statistically significant (P<0.05). By multivariate logistic analysis, the large area cerebellar infarction (P<0.05), pre-op thrombolysis(P<0.01), pre-op mild HT (P<0.01), oral anticoagulants (P<0.01) were the independent risk factors for post-op HT. Conclusions: The large area cerebellar infarction (the diameter of area was more than 5 cm), pre-op thrombolysis, pre-op mild HT, oral anticoagulants, atrial fibrillation, hyperglycemia and fluctuation of BP in post-op are important risk factors for post-op HT. The large area cerebellar infarction, pre-op thrombolysis, pre-op mild HT, oral anticoagulants are the independent risk factors for post-op HT. A proper pre-op evaluation of these risk factors and an individualized treatment for post-op HT would help a lot with balancing operational risk and improving prognosis.

Long non-coding RNA HOTAIR regulates the development of non-small cell lung cancer through miR-217/DACH1 signaling pathway.

OBJECTIVE: Long non-coding RNA HOX transcript antisense RNA (HOTAIR) is reported to make chromatin state, cell growth and cancer metastasis. However, the role of HOTAIR in non-small cell lung cancer (NSCLC) remains unknown. The aim of this study was to explore the regulatory mechanism of HOTAIR in NSCLC in relation to miR-217/Dachshund homolog 1 (DACH1) signaling pathway. MATERIALS AND METHODS: The expression levels of HOTAIR and miR-217 were measured by quantitative Polymerase Chain Reaction (qPCR) in NSCLC cell lines and human bronchial epithelial cell line HBE. The direct target of HOTAIR and miR-217 in NSCLC was confirmed by a Luciferase reporter assay. The expression of DACH1 protein was examined by Western blot (WB) assay. Cell migration and invasion were examined with transwell assays, and cell proliferation was measured by Cell Counting Kit-8 (CCK8) assay. RESULTS: HOTAIR was up-regulated and miR-217 was down-regulated in NSCLC cell lines. Silencing of HOTAIR significantly repressed cell proliferation and inhibited cell migration and invasion in H1299 and A549 cells by facilitating miR-217 expression. Moreover, bioinformatics analysis and Luciferase reporter assay confirmed that DACH1 was a target of miR-217. Furthermore, the overexpression of miR-217 markedly repressed cell proliferation and inhibited cell migration and invasion in H1299 and A549 cells. DACH1 reverses the effects of miR-217 overexpression in NSCLC cells. CONCLUSIONS: HOTAIR was up-regulated in NSCLC cell and regulates the proliferation, migration, invasion through the miR-217/DACH1 signaling pathway. It provides a novel potential treatment strategy for NSCLC.

CARD9 mutation linked to Corynespora cassiicola infection in a Chinese patient.

Corynespora cassiicola is a plant pathogen associated with leaf-spotting disease. The fungus has been found on diverse substrates: leaves, stems and roots of plants; nematode cysts and human skin. It rarely causes human infections. Here we report one case of subcutaneous phaeohyphomycosis caused by C. cassiicola with prominent tissue necrosis in a woman. All of her clinical features pointed towards a genetic linkage. Hence, whole-exome sequencing and Sanger sequencing were performed on this patient. One mutation of CARD9 was detected.

Specific Cleavage of the Nucleoprotein of Fish Rhabdovirus.

Siniperca chuatsi rhabdovirus (SCRV) is one of myriad rhabdoviruses recorded in fish. Preliminary data show that inhibition of the SCRV nucleoprotein (N) could significantly reduce the progeny virus titers in infected Epithelioma papulosum cyprinid (EPC) cells. Here, the authors propose that cleavage of the viral 47-kDa N protein is caspase-mediated based on caspase inhibition experiments, transient expression in EPC transfection, and analysis of cleavage sites. Cleavage of the SCRV N protein in culture was prevented by a pan-caspase inhibitor, z-VAD-FMK (z-Val-Ala-DL-Asp-fluoromethyl ketone). Subsequently, N was transiently expressed in EPC cells, the results of which indicated that the specific cleavage of N also occurred in the cells transfected with N-GFP plasmid. Several truncated fragments of the N gene were constructed and transiently transfected into EPC cells. Immunoblotting results indicated that D324 and D374 are the cleavage sites of N by caspases. The authors also found that z-VAD-FMK could inhibit the cytopathic effect in SCRV-infected EPC cells but not affect the production of infectious progeny, suggesting that the caspase-mediated cleavage of N protein is not required for in vitro SCRV replication. To the authors' knowledge, this is the first report on the cleavage of rhabdovirus proteins.

Mechanism of curcumin analog MHMD-induced cell death in A549 lung cancer cells.

OBJECTIVE: To investigate the anticancer properties of a chemosynthetic curcumin analog, (1E,6E)-4-((furan-2-yl)methylene)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (C26H22O7, abbreviated MHMD) in A549 cells. MATERIALS AND METHODS: Inverted microscope was used to observe the alteration on cytomorphology. MTT assay was used to detect cell viability. Acridine-orange staining was used to measure autophagy, and AnnexinV/PI staining and Hoechst/PI staining to measure apoptosis and necrosis. RESULTS: MTT assays showed that at 12 h, 24 h, 48 h, MHMD reduced cell viability with an IC50 of 27.46 µM, 18.86 µM, and 11.23 µM, respectively. Typical characteristics were observed in concert with cell death, including treated-cells getting brighter, rounder, and becoming non-adherent gradually. Additionally, acridine-orange staining suggested that autophagy didn't involve in MHMD-induced cell death. However, apoptosis and necrosis played important roles in MHMD-induced cell death by Hoechst33342/PI staining. It showed apoptosis was the main cause at low concentrations (≤ 4 µM), while with the concentrations rising, necrosis was the leading role. AnnexinV/PI staining again indicated the occurrence of apoptosis at 4 µM. Furthermore, the caspases inhibitor z-VAD-fmk could prevent MHMD-induced cell death, which showed much higher cell viability than those only treated with MHMD (4 µM). Moreover, MTT assay also demonstrated that MHMD did possess a greater anti-proliferative ability than curcumin. CONCLUSIONS: The curcumin analog MHMD is able to induce A549 cell death in a time and dose-dependent manner via apoptosis and necrosis. And MHMD could be a more effective drug than curcumin.

Student and intern awareness of ionising radiation exposure from common diagnostic imaging procedures.

This study aims to evaluate medical student and intern awareness of ionising radiation exposure from common diagnostic imaging procedures and to suggest how education could be improved. Fourth to sixth year medical students enrolled at a Western Australian university and interns from three teaching hospitals in Perth were recruited. Participants were asked to complete a questionnaire consisting of 26 questions on their background, knowledge of ionising radiation doses and learning preferences for future teaching on this subject. A total of 331 completed questionnaires were received (95.9%). Of the 17 questions assessing knowledge of ionising radiation, a mean score of 6.0 was obtained by respondents (95% CI 5.8-6.2). Up to 54.8% of respondents underestimated the radiation dose from commonly requested radiological procedures. Respondents (11.3 and 25.5%) incorrectly believed that ultrasound and MRI emit ionising radiation, respectively. Of the four subgroups of respondents, the intern doctor subgroup performed significantly better (mean score 6.9, P < 0.0001, 95% CI 6.5-7.3) than each of the three medical student subgroups. When asked for the preferred method of teaching for future radiation awareness, a combination of lectures, tutorials and workshops was preferred. This study has clearly shown that awareness of ionising radiation from diagnostic imaging is lacking among senior medical students and interns. The results highlight the need for improved education to minimise unnecessary exposure of patients and the community to radiation. Further studies are required to determine the most effective form of education.

Lower CSF level of dynorphin(1-8) immunoreactivity in schizophrenic patients.

Using a highly specific and sensitive radioimmunoassay for dynorphin(1-8) (D 1-8), and a singly blind test design, we measured D(1-8) immunoreactivity (ir D 1-8) in CSF of 35 first break cases of acute schizophrenic patients. All patients were free of psychotropic medication for at least one week before the study. Another 31 neurological patients suffered from cervical arthrosis, tumor, myelopathy etc. were studied as controls. The ir D(1-8) in CSF of schizophrenic patients were significantly lower than that of controls (91.8 +/- 5.6, means +/- S.E.M., and 131.9 +/- 6.8 fmol/ml CSF respectively, p less than 0.001).