患者再現videoを利用したPBLテュートリアルの有用性に関する研究

研究科: 千葉大学大学院医学薬学府（先端医学薬学専攻）学位記番号: 千大院医薬博甲第医1338号博士（医学）千葉大学 = Chiba Universit

Wing images

This file is a .zip archive containing images of wings from butterflies with presumptive spalt and Distal-less mosaic gene deletion phenotypes. A reference table is included

Small motif architecture of <i>VDAC2</i> in <i>Crassostrea gigas</i>.

<p>Motif analysis of <i>CgVDAC2</i>. <i>CgVDAC2</i> contains a 4-element eukaryotic porin signature motif (box with diagonal lines), a glycine-leucine-lysine (GLK) motif (black box), and a VKAKV-like sequence (box with horizontal lines). The four-element eukaryotic porin signature motif is indicated above. The nucleotide and amino acid number of the start site of each motif is labeled below.</p

Neighbor-joining phylogenetic tree of <i>VDAC1</i>s and <i>VDAC2</i>s from different vertebrate and invertebrate species.

<p>The neighbor-joining tree which was built by MEGA program was based on the sequences of <i>CgVDAC2</i> from <i>Crassostrea gigas</i>, and <i>VDAC2</i>s from other species, including <i>Homo sapiens</i> (NP_003366), <i>Mus musculus</i> (NP_035825), <i>Xenopus laevis</i> (NP_001089399), <i>Danio rerio</i> (NP_955879), <i>Aplysia californica</i> (XP_005113380), <i>Haliotis diversicolor</i> (ADI56517), <i>Drosophila melanogaster</i> (NP_609462), <i>Lepeophtheirus salmonis</i> (ADD24283), <i>Nematostella vectensis</i> (XP_001623935), <i>Hydra vulgaris</i> (XP_002167561). Besides, four VDAC1 sequences from <i>Homo sapiens</i> (NP_003365), <i>Mus musculus</i> (NP_035824), <i>Xenopus laevis</i> (NP_001080684), and <i>Danio rerio</i> (NP_001001404) were also used in the phylogenetic analysis.</p

Subcellular localization of CgVDAC2-EGFP in HeLa cells.

<p>The plasmids of <i>CgVDAC2</i> and the enhanced green fluorescent protein (EGFP) negative control were transfected into HeLa cells (green). Cell nuclei are stained with Hoechst 33342 (blue) and cell membranes with Alexa Fluor 594 (red). The green fluorescent signal of CgVDAC2-EGFP is mainly localized to cytoplasmic puncta and indicates the co-localization of CgVDAC2 with mitochondria in HeLa cells. Scale bars = 5 μm.</p

Expression pattern of <i>CgVDAC2</i> transcript in ostreid herpesvirus 1 (OsHV-1) infected hemolymph.

<p>Quantitative PCR analysis of <i>CgVDAC2</i> expression in <i>C</i>. <i>gigas</i> at 0, 3, 6, 12, 24, 48 and 72 h after treatment with OsHV-1 (infected samples) or phosphate-buffered saline (PBS, control). Samples at 0 h after both treatments were used as the reference samples. The experiments were repeated three times. The values are shown as the mean ± S.D (N = 3). Asterisks indicate significant differences at <i>P</i> < 0.05 * and <i>P</i> < 0.01**.</p

Effects of overexpression of CgVDAC2 on UV irradiation-induced apoptosis in HEK293T cells.

<p>(A) Recombinant expression of CgVDAC2 and CgBak in HEK293T cells. The deduced molecular weights of these two proteins are approximate 30 kDa and 26 kDa, respectively. The asterisk indicated a non-specific band. (B) Caspase3 activities of HEK293T cells that expressed distinct recombinant proteins. The Caspase3 activities were determined 24 h after UV irradiation and based on spectrophotometric detection of the chromophore <i>p</i>-nitroaniline (<i>p</i>NA) after cleavage from the labeled substrate DEVD-<i>p</i>NA. The values are shown as the mean ± S.D (N = 3). Different small letters indicate differences (<i>P</i> < 0.05) and the same letter indicated not.</p

Effects of RNAi of <i>CgVDAC2</i> on UV irradiation-induced apoptosis in hemocytes of <i>C</i>.<i>gigas</i>.

<p>(A) Expression level of <i>CgVDAC2</i> in hemocytes of <i>C</i>.<i>gigas</i> after RNAi. Error bars represent standard error of three parallels. Different small letters indicate differences at <i>P</i> < 0.05. (B) Apoptosis level of hemocytes with RNAi-silenced <i>CgVDAC2</i> upon UV light irradiation. Values represent the mean ± SD of six samples. Different small letters indicate differences at <i>P</i> < 0.05.</p

Developmental stage distributions and tissue distributions of <i>CgVDAC2</i> transcripts.

<p>(A) <i>CgVDAC2</i> mRNA expression pattern at 11 different developmental stages of <i>Crassostrea gigas</i>. Data are displayed as the mean ± standard error of triplicate independent experiments. E, egg stage; TC, two-cell stage; FC, four-cell stage; M, morula stage; B, blastula stage; G, gastrula stage; T, trochophore stage; ED, early D-shape larval stage; LD, late D-shape larval stage; EU, early umbo larval stage; LU, late umbo larval stage. Significant differences between the expression levels in each developmental stage and those in the former stage were identified using <i>t</i>-test. Asterisks indicate significant differences at <i>P</i> < 0.05 * and <i>P</i> < 0.01**. <b>(</b>B) <i>CgVDAC2</i> mRNA expression pattern in eight different tissues of <i>Crassostrea gigas</i>. <i>CgVDAC2</i> expression in mantle, adductor muscle, labial palp, gill, male gonad, digestive gland, hemolymph and female gonad are shown. Data are displayed as the mean ± standard error of triplicate independent experiments. Asterisks indicate significant differences at <i>P</i> < 0.05 * and <i>P</i> < 0.01**.</p

Image_1_Next-generation sequencing of homologous recombination genes could predict efficacy of platinum-based chemotherapy in non-small cell lung cancer.tif

BackgroundWith the widespread use of next-generation sequencing (NGS) in clinical practice, an increasing number of biomarkers that predict a response to anti-tumor therapy in non-small cell lung cancer (NSCLC) has been identified. However, validated biomarkers that can be used to detect a response to platinum-based chemotherapy remain unavailable. Several studies have suggested that homologous recombination deficiency (HRD) may occur in response to platinum-based chemotherapy in ovarian cancer and breast cancer. However, currently there is a lack of proven and reliable HRD markers that can be used to screen for patients who may benefit from platinum-based chemotherapy, especially in NSCLC.MethodsNGS was used to screen for gene mutations, including homologous recombination (HR) genes and common driver gene mutations in NSCLC. Cox regression analysis was performed to identify potential clinicopathological or gene mutation factors associated with survival in patients receiving platinum-based chemotherapy, while Kaplan–Meier analysis with the log-rank test was performed to assess the effect of HR gene mutations on progression-free survival (PFS).ResultsIn a retrospective cohort of 129 patients with advanced NSCLC, 54 who received platinum-based chemotherapy with or without anti-angiogenic therapy were included in the analysis. Univariate and multivariate Cox proportional hazard regression analyses showed that HR gene mutations were associated with platinum-based chemotherapy sensitivity. Efficacy results indicated that the objective response rates (ORR) for patients with BRCA1/2 mutations and BRCA1/2 wild type were 75% and 30.4% (p=0.041), while the median PFS was 7.5 and 5.5 months (hazard ratio [HR], 0.52; 95% CI, 0.27–1.00; p=0.084), respectively. The ORRs of patients with HR gene mutations and HR gene wild type were 60% and 23.6% (p=0.01), and the median PFS was 7.5 and 5.2 months (HR, 0.56; 95% CI, 0.32–0.97; p=0.033), respectively.ConclusionsHR gene mutations show potential as promising biomarkers that may predict sensitivity to platinum-based chemotherapy in advanced and metastatic NSCLC.</p