DataSheet_2_The two-directional prospective association between inflammatory bowel disease and neurodegenerative disorders: a systematic review and meta-analysis based on longitudinal studies.doc

ObjectivePrevious studies reported possible connections between inflammatory bowel disease (IBD) and several neurodegenerative disorders. However, the comprehensive relationships between IBD and various neurodegenerative disorders were not summarized. We executed a meta-analysis of longitudinal studies to provide an estimate of the strength of the two-directional prospective association between IBD and neurodegenerative disorders.MethodsWe accomplished a thorough bibliographic search of PubMed, Web of Science, Embase, PsycINFO, and Cochrane Library databases until June 2023 to locate relevant longitudinal studies. The extracted data were then analyzed via meta-analysis using either a fixed or random effects model.ResultsThe final analysis encompassed 27 studies. Individuals with IBD faced an increased risk of developing four neurodegenerative disorders than the general public, namely, Alzheimer’s disease (hazard ratio[HR] = 1.35, 95% confidence interval [CI]: 1.03–1.77, P=0.031), dementia (HR =1.24, 95% CI: 1.13–1.36, PInterpretationThese findings verified the two-directional relationship between the brain-gut axis, specifically demonstrating a heightened risk of various neurodegenerative diseases among IBD patients. It may be profitable to prepare screening strategies for IBD patients to find neurodegenerative diseases during the long-term course of treatment for IBD with a view to potential earlier diagnosis and treatment of neurodegenerative diseases, reducing public health and social burden.Systematic Review RegistrationPROSPERO (CRD42023437553).</p

DataSheet_1_The two-directional prospective association between inflammatory bowel disease and neurodegenerative disorders: a systematic review and meta-analysis based on longitudinal studies.doc

ObjectivePrevious studies reported possible connections between inflammatory bowel disease (IBD) and several neurodegenerative disorders. However, the comprehensive relationships between IBD and various neurodegenerative disorders were not summarized. We executed a meta-analysis of longitudinal studies to provide an estimate of the strength of the two-directional prospective association between IBD and neurodegenerative disorders.MethodsWe accomplished a thorough bibliographic search of PubMed, Web of Science, Embase, PsycINFO, and Cochrane Library databases until June 2023 to locate relevant longitudinal studies. The extracted data were then analyzed via meta-analysis using either a fixed or random effects model.ResultsThe final analysis encompassed 27 studies. Individuals with IBD faced an increased risk of developing four neurodegenerative disorders than the general public, namely, Alzheimer’s disease (hazard ratio[HR] = 1.35, 95% confidence interval [CI]: 1.03–1.77, P=0.031), dementia (HR =1.24, 95% CI: 1.13–1.36, PInterpretationThese findings verified the two-directional relationship between the brain-gut axis, specifically demonstrating a heightened risk of various neurodegenerative diseases among IBD patients. It may be profitable to prepare screening strategies for IBD patients to find neurodegenerative diseases during the long-term course of treatment for IBD with a view to potential earlier diagnosis and treatment of neurodegenerative diseases, reducing public health and social burden.Systematic Review RegistrationPROSPERO (CRD42023437553).</p

CCR4 expression levels in human gastric cancer cells and non-cancerous gastric tissues.

<p>CCR4 was detected at different levels, including negative expression (-) (A), weak expression (+) (B), moderate expression (++) (C) and strong expression (+++) (D). The expression was negative in non-neoplastic cells (the right area in the image) adjacent to CCR4 positive gastric cancer cells (E), and weak expression of membrane CCR4 was found in gastric preneoplastic lesions (F). Magnification, ×400 (A-D, F), ×200 (E).</p

Relationship between CCR4 expression and invasive behaviors of gastric cancer cells.

<p>CCR4 expression was low in less aggressive and well-differentiated cancer cells (A), whereascytoplasmic CCR4 expression was high in highly invasive cancer cells, including those with muscle invasion (B), vascular penetration (C) and nerve invasion (D). Magnification, ×400.</p

Aberrant CCR4 Expression Is Involved in Tumor Invasion of Lymph Node-Negative Human Gastric Cancer

<div><p>Cellular chemotaxis is the best-known function of chemokine receptors which are closely linked with tumor metastasis. In fact, positive expression of chemokine receptors could also be identified even in some patients without metastatic tumors, while the clinical relevance of this data has not been fully established. Our studies were designed to clarify the CCR4 expression profiles and to explore its potential role in histologically node-negative (pN0) gastric cancer (GC). Immunohistochemistry (IHC) or immunohistofluorescence (IHF) analysis was performed on specimens obtained from 108 patients with pN0 GC. We found that CCR4 was aberrantly over-expressed inpN0 GC tissues, with different expression patterns on tumor cells and being associated with T-stage (<i>P</i> = 0.002). The matrigel <i>in vitro</i> invasion assay revealed that over-expression of CCR4 in GC cell lines significantly enhanced the invasive capacity of these cells. Results from real-time RT-PCR and gelatinzymography showed a significant increase in matrix metalloproteinase (MMP)-9 production induced by the forced expression of CCR4 in GC cell lines. Our data suggest that the aberrant CCR4 expression is involved in tumor invasion of pN0 GC and, conceivably, antagonists of CCR4 might be useful candidates for controlling early events in tumor progression.</p></div

Over-expression of CCR4 up-regulated MMP-9 production in human GC cells.

<p>A, real time RT-PCR showed that MMP-9 mRNA expression was upregulated in CCR4-transfected GC cells compared to that in MOCK-transfected cells at 24h after transfection. B, zymograms showing gelatin digestion by pro-MMP-9 (92 kd) and active MMP-9 (85 kd). The secretion of MMP-9 were clearly up-regulated in the supernatants of CCR4-transfected cells compared with MOCK-transfected cells at 48h after transfection, with or without the addition of exogenous CCL22. Data are presented as representative images or mean ± SD and for three separate experiments from each group. *, <i>P</i>< 0.05; **, <i>P</i>< 0.01.</p

Distinct expression patterns of CCR4 in human gastric cancer.

<p>Green fluorescence represents positive cells with membrane and cytoplasmic distribution of CK, red fluorescence represents CCR4-positive cells, whereas yellow represents the merge of both. And both membrane and cytoplasm expression of CCR4 were observed most frequently in well-differentiated tumor nests of intestinal type, while the less-differentiated cells tended to display a cytoplasmic pattern of CCR4 expression in diffuse type gastric cancer. Magnification, ×400.</p

Relation between CCR4 expression and clinical characteristics of pN0 gastric cancer.

<p>¹Chi-squared test.</p><p>²Mean age of cases is 62 years.</p><p>Relation between CCR4 expression and clinical characteristics of pN0 gastric cancer.</p

Over-expression of CCR4 enhanced the invasion of human GC cells.

<p>SGC-7901 and BGC-823 cells were transiently transfected with empty (MOCK) or CCR4-expressing vector (EGFP-CCR4) using lipofectamine2000, and the transfection efficiency was confirmed by a fluorescence microscope (A), RT-PCR (B) and FCM (C) at 24 or 48 hours after transfection, respectively. D, cells were resuspended in 200μl serum-free RPMI-1640 with a concentration of 1.5×10⁵/ml after transfection at 24h, and then allowed to invade the Transwell inserts (8-μm pores) coated with Matrigel for another 24h. The cells that invaded through the inserts were counted and photographed under a light microscopy at 200× magnification. The data are presented as representative images or as the mean ± SD (n = 3 in each column) from more than three individual experiments. Gray-filled histogram, CCR4; empty histogram, isotype. Data are presented as representative images and mean ± SD and for three separate experiments from each group. *, <i>P</i>< 0.05; **, <i>P</i>< 0.01.</p

Schematic procedure of virus in-capsule-mucus penetration system.

<p>(1) 150 µl of mucus were brought at the bottom of a gelatin capsule. (2) 8 µl of SIV suspension were added on top of the surface of the mucus. (3) Mucus together with virus was embedded and snap-frozen. (4) Cryosections were made vertically to the mucus surface. (5) Immunofluorescence staining was performed to visualize the Muc5AC (representing the mucus) and viral particles. (6) Penetration depth (shown by yellow arrows) was measured from the surface of mucus to the furthest point of the viral signal.</p