Hcrt neuronal number is increased in dark, but cell size and anatomical distribution are unchanged.

<p>(A) Average number of Hcrt cells in the whole hypothalamus. (B) Total cell counts for each experimental subject. (C) Average size of Hcrt neurons. (D) Distribution of Hcrt neurons in different hypothalamic subdivisions. (D) (insert), Diagram of one hypothalamic section of an experimental animal sacrificed during the light phase indicating the anatomical subdivision used, circles represent Hcrt neurons (purple, medial hypothalamus; brown, lateral hypothalamus; blue, perifornical area). (E) Photomicrographs of the same hypothalamic area of an animal sacrificed in the light phase (ZT5, top) and an animal sacrificed in the dark phase (ZT17, bottom). Numbers indicate cell counts on slide. Inserts are higher (x40) magnification photomicrographs of the selected area (black square) for each experimental condition. Calibration bar 250 μm and 10 μm respectively. Fx, fornix; LH, lateral hypothalamus; MH, medial hypothalamus; PFA, perifornical area; 3V third ventricle. ** p = 0.02, Wilcoxon-Mann-Whitney, n = 4 for each group.</p

Number and size of ChAT neurons does not vary between the light and dark phase.

<p>(A) Average number of ChAT cells in the horizontal diagonal band. n = 4 for each group. (B) Average size of ChAT neurons.</p

Number of neurons during the light and dark phase.

<p>Number of neurons during the light and dark phase.</p

Number of MCH neurons remains unchanged, but cell size is increased in the light.

<p>(A) Average number of MCH cells in the light and dark phases. (B) Distribution of MCH neurons in different hypothalamic subdivisions. (C) Average size of MCH neurons. ** p = 0.02, Wilcoxon-Mann-Whitney, n = 4 for each group. LH, lateral hypothalamus; MH, medial hypothalamus; PFA, perifornical area</p

Pre-CS conditioned freezing behaviour is significantly higher in 5-HTT^−/− rats compared to 5-HTT^+/+ rats regardless of housing conditions.

<p>(A) The inset figure shows increased pre-CS freezing in 5-HTT^−/− rats compared to 5-HTT^+/+rats housed in open cages (B). Data represent mean ± S.E.M. * P<0.05, ** P<0.01.</p

Primers used for qPCR analysis.

<p>Primers used for qPCR analysis.</p

Colchicine further increases the number of Hcrt neurons.

<p><b>MCH and ChAT neuronal numbers remain unchanged</b>. (A) Photomicrographs of the same hypothalamic area of an animal treated with saline (top) or colchicine (bottom). Calibration bar 250 μm. Insert corresponds to a higher (x60) magnification of the selected area (black square) of the animal that received colchicine. Black arrow indicates an Hcrt neuron. Calibration bar 10 μm. (B) Total cell counts for each experimental subject. (C) Average number of Hcrt neurons in animals with ICV saline vs. ICV colchicine injections (** p = 0.02, Wilcoxon-Mann-Whitney, n = 5 for each group). (D) Distribution of Hcrt neurons in the three different hypothalamic subdivisions regions. Increase is greatest in the medial hypothalamic region (*p<0.05, Wilcoxon-Mann-Whitney with Bonferroni correction). (E) relative PPHcrt mRNA expression in the hypothalamus. The difference is not significant. (F) Average number of MCH neurons in animals with ICV saline vs. ICV colchicine injections. (G) Relative PPMCH mRNA expression in the hypothalamus (** p = 0.02, Wilcoxon-Mann-Whitney, n = 8 for each group). (H) Diagram of the basal forebrain of the mouse depicting the anatomical localization of the cholinergic cell group analyzed. Lower, photomicrographs of the same basal forebrain area of an animal treated with saline (above) or colchicine (below). In contrast to the increase in the staining of the Hcrt and MCH peptide, staining of ChAT is not increased by colchicine treatment. Calibration bar, 250 μm. (I) Average number of ChAT neurons in animals with ICV saline vs. ICV colchicine injections. LH, lateral hypothalamus; MH, medial hypothalamus; PFA, perifornical area; Fx, fornix; HDB, horizontal diagonal band; VP, ventral pallidum.</p

Conditioned freezing behaviour as observed during the fear extinction session conducted 24 hours after fear conditioning.

<p>Freezing behaviour was impaired in 5-HTT^−/− (n = 10) compared to 5-HTT^+/+ (n = 10) rats housed in open cages (A); The replication experiment also showed impaired fear extinction in 5-HTT^−/− (n = 5) <i>versus</i> 5-HTT^+/+ (n = 5) rats housed in open cages (B); Lack of differences in freezing behaviour across trials between 5-HTT^−/− (n = 7) and 5-HTT^+/+ (n = 8) rats housed in IVC cages (C); This lack of differences in freezing behaviour was confirmed in an independent replication experiment consisting of five 5-HTT^−/− and five 5-HTT^+/+ rats housed in IVC cages (D). Data (trial blocks averaged over 3 trials) represent mean percentage of freezing time (± S.E.M.) during CS presentation. * P<0.05.</p

Detection of <i>ROS1</i> Gene Rearrangement in Lung Adenocarcinoma: Comparison of IHC, FISH and Real-Time RT-PCR

<div><p>Aims</p><p>To compare fluorescence <i>in situ</i> hybridization (FISH), immunohistochemistry (IHC) and quantitative real-time reverse transcription-PCR (qRT-PCR) assays for detection of <i>ROS1</i> fusion in a large number of <i>ROS1</i>-positive lung adenocatcinoma (ADC) patients.</p><p>Methods</p><p>Using IHC analysis, sixty lung ADCs including 16 cases with ROS1 protein expression and 44 cases without ROS1 expression were selected for this study. The <i>ROS1</i> fusion status was examined by FISH and qRT-PCR assay.</p><p>Results</p><p>Among 60 cases, 16 (26.7%), 13 (21.7%) and 20 (33.3%) cases were <i>ROS1</i> positive revealed by IHC, FISH and qRT-PCR, respectively. Using FISH as a standard method for <i>ROS1</i> fusion detection, the sensitivity and specificity of IHC were 100% and 93.6%, respectively. Three IHC-positive cases, which showed FISH negative, were demonstrated with <i>ROS1</i> fusion by qRT-PCR analysis. The sensitivity and specificity of qRT-PCR for detection for <i>ROS1</i> fusion were 100% and 85.1%, respectively. The total concordance rate between IHC and qRT-PCR were 93.3%.</p><p>Conclusion</p><p>IHC is a reliable and rapid screening tool in routine pathologic laboratories for the identification of suitable candidates for ROS1-targeted therapy. Some ROS1 IHC-positive but FISH-negative cases did harbor the translocation events and may benefit from crizotinib.</p></div

Increased Incidence of Thyroid Disease in Patients with Celiac Disease: A Systematic Review and Meta-Analysis

<div><p>The prevalence of thyroid disease is likely increased among individuals with celiac disease (CD). In addition, exposure to gluten-free treatment may be associated with a risk of thyroid disease, but this association remains controversial. A systematic review was performed to evaluate the association between thyroid disease and CD. The articles were obtained from the PubMed, Web of Science, Embase, and Chinese WanFang bibliographical databases for the period up to May 2016. The results were analysed in a meta-analysis with odds ratios (ORs) and corresponding 95% confidence intervals (95% CIs). There were 13 articles in this meta-analysis, including 15629 CD cases and 79342 controls. Overall, the prevalence of thyroid disease in patients with CD was significantly increased compared with that in the control groups (OR 3.08, 95% CI 2.67–3.56, <i>P</i><0.001). Moreover, there was no significant difference in the OR between the gluten-treated and untreated groups (OR 1.08, 95% CI 0.61–1.92, <i>P</i> = 0.786). The results of our meta-analysis support the hypothesis that the prevalence of thyroid disease in patients with CD is increased compared with that in controls, which suggests that CD patients should be screened for thyroid disease. The effect of gluten-free treatment on thyroid disease needs further investigation.</p></div