Identification of immune-related biomarkers in peripheral blood of schizophrenia using bioinformatic methods and machine learning algorithms.

Schizophrenia is a group of severe neurodevelopmental disorders. Identification of peripheral diagnostic biomarkers is an effective approach to improving diagnosis of schizophrenia. In this study, four datasets of schizophrenia patients' blood or serum samples were downloaded from the GEO database and merged and de-batched for the analyses of differentially expressed genes (DEGs) and weighted gene co-expression network analysis (WCGNA). The WGCNA analysis showed that the cyan module, among 9 modules, was significantly related to schizophrenia, which subsequently yielded 317 schizophrenia-related key genes by comparing with the DEGs. The enrichment analyses on these key genes indicated a strong correlation with immune-related processes. The CIBERSORT algorithm was adopted to analyze immune cell infiltration, which revealed differences in eosinophils, M0 macrophages, resting mast cells, and gamma delta T cells. Furthermore, by comparing with the immune genes obtained from online databases, 95 immune-related key genes for schizophrenia were screened out. Moreover, machine learning algorithms including Random Forest, LASSO, and SVM-RFE were used to further screen immune-related hub genes of schizophrenia. Finally, CLIC3 was found as an immune-related hub gene of schizophrenia by the three machine learning algorithms. A schizophrenia rat model was established to validate CLIC3 expression and found that CLIC3 levels were reduced in the model rat plasma and brains in a brain-regional dependent manner, but can be reversed by an antipsychotic drug risperidone. In conclusion, using various bioinformatic and biological methods, this study found an immune-related hub gene of schizophrenia - CLIC3 that might be a potential diagnostic biomarker and therapeutic target for schizophrenia.

The SIK1/CRTC2/CREB1 and TWIST1/PI3K/Akt/GSK3ß signaling pathways mediated by microRNA-25-3p are altered in the schizophrenic rat brain.

Schizophrenia is a group of severe mental disorders. MiR-25-3p was shown to be involved in various neuropsychiatric diseases and can regulate SIK1 and TWIST1. The CRTC2/CREB1 and PI3K/Akt/GSK3ß signaling pathways are downstream pathways of SIK1 and TWIST1, respectively. This study investigated whether miR-25-3p-mediated SIK1/CRTC2/CREB1 and TWIST1/PI3K/Akt/GSK3ß signaling pathways are present in an animal model relevant to schizophrenia. A schizophrenic rat model was established by using sub-chronic MK-801 administration. An RNA-seq test was performed to examine the differentially expressed genes (DEGs) in the rat prefrontal cortex (PFC). The mRNA levels of miR-25-3p, SIK1, and TWIST in the PFC and caudate putamen (CPu) were assessed by qRT-PCR. Phosphorylation of the SIK1/CRTC2/CREB1 and TWIST1/PI3K/Akt/GSK3ß pathways in the two brain regions was examined by Western blots. The RNA-seq data revealed down-regulated miR-25-3p expression and up-regulated SIK1 and TWIST1 mRNA expression induced by MK-801. Additionally, SIK1 and TWIST1 were shown to be possible downstream responders of miR-25-3p in previous studies. qRT-PCR confirmed the changes of miR-25-3p, SIK1, and TWIST1 induced by MK-801 in both brain regions, which, however, was reversed by risperidone. Furthermore, the phosphorylation of the SIK1/CRTC2/CREB1 pathway was repressed by MK-801, whereas the phosphorylation of the TWIST1/PI3K/Akt/GSK3ß pathway was increased by MK-801 in either of the two brain regions. Moreover, the altered phosphorylation of these two signaling pathways induced by MK-801 can be restored by risperidone. In conclusion, this study suggests that altered SIK1/CRTC2/CREB1 and TWIST1/PI3K/Akt/GSK3ß signaling pathways mediated by miR-25-3p is very likely to be associated with schizophrenia, revealing potential targets for the treatment and clinical diagnosis of schizophrenia.

Dysfunctional microRNA-144-3p/ZBTB20/ERK/CREB1 signalling pathway is associated with MK-801-induced schizophrenia-like abnormalities.

Schizophrenia is a group of severe mental disorders. ZBTB20 is critical in brain development and corticogenesis and its dysfunction induces various neural disorders. ERK/CREB signalling is a potential downstream pathway of ZBTB20. Up-regulated microRNA-144-3p (miR-144-3p) were found in schizophrenic model rats in our previous study. This study investigated whether suppressed ZBTB20/ERK/CREB1 signalling caused by up-regulated miR-144-3p is associated with schizophrenia-like abnormalities in animals. A MK-801 rat model was established by 2-week MK-801 administration. An RNA-Seq test was performed to reveal differentially expressed genes in model rat hippocampus (HIP). MiR-144-3p-overexpressed SK-N-SH and 293T cell models were constructed by lentivirus, respectively. The in vitro and in vivo levels of miR-144-3p and ZBTB20, and the activation of the ERK/CREB1 signalling were examined by qRT-PCR, Western blots, or immunohistochemistry. The interaction between miR-144-3p and ZBTB20 was predicted and assessed by using bioinformatic methods and a luciferase reporter gene assay on 293T cells, respectively. The RNA-Seq test revealed that ZBTB20 was altered in the model rat HIP. Further experiments confirmed the reduced ZBTB20 mRNA and protein levels in the model rat HIP and caudate putamen (CPu), accompanied by increased miR-144-3p levels. Moreover, the ZBTB20 expression and ERK/CREB1 phosphorylation was decreased in the miR-144-3p-overexpressed 293T cells. These abnormal changes in the ZBTB20 expression and ERK/CREB1 phosphorylation levels were also observed in the model rat brain, but could be reversed by risperidone. In conclusion, this study revealed that dysfunctional miR-144-3p/ZBTB20/ERK/CREB1 signalling might be associated with schizophrenia-like abnormalities, suggesting potential therapeutic targets for future schizophrenia treatment.

Effects of icariin on alleviating schizophrenia-like symptoms by regulating the miR-144-3p/ATP1B2/mTOR signalling pathway.

Schizophrenia is a group of severe mental disorders. Icariin is a main active component of Epimedium, possessing therapeutic effects on various neurodegenerative diseases. The present study investigated whether icariin is effective in alleviating schizophrenia-like symptoms and explored its underlying molecular mechanism. A developmental schizophrenia rat model employing 2-week repeated MK-801 administration was established. Icariin was orally administrated 3 time per day for 2 weeks after the MK-801 administration. Open-field test (OFT), novel object recognition (NOR), rotarod, and Morris water maze (MWM) were performed to examine the therapeutic effects of icariin on behavioural abnormalities. Hematoxylin-eosin (HE) staining on hippocampus slices, and MTT assay and Calcein/PI staining on the SK-N-SH cells treated with MK-801 were carried out to assess the neuroprotective effects of icariin. Furthermore, the regulation of icariin on the miR-144-3p/ATP1B2/mTOR signalling pathway was examined by RT-PCR and Western blots. The results showed that icariin alleviated MK-801-induced anxiety and recognition memory deficits in the OFT and NOR, respectively. Additionally, weakened motor coordination caused by MK-801 was restored by icariin. The MWM test also showed that icariin can improve MK-801-induced impaired spatial memory and swimming ability. Furthermore, brain grey matter atrophy, cytotoxicity, and cell apoptosis caused by MK-801 can be eliminated by icariin. Lastly, icariin can regulate the expression of miR-144-3p and ATP1B2, and enhance the phosphorylation of PI3K, Akt, and mTOR. In conclusion, this study revealed that icariin may have therapeutic effects on schizophrenia-like disorders via regulating the miR-144-3p/ATP1B2/mTOR signalling, suggesting that icariin has potential to become an antipsychotic drug.

Efficacy and Safety of Single-Balloon Versus Double-Balloon Enteroscopy: A Single-Center Retrospective Analysis.

BACKGROUND Single-balloon endoscopy (SBE) has been introduced as a simplified endoscopy technique after the promotion of double-balloon endoscopy (DBE). The difference in clinical performance between DBE and SBE is still not very clear. In this study, we aimed to compare the efficacy and safety between these 2 endoscopic procedures. MATERIAL AND METHODS A total of 173 patients with suspected small bowel disease were enrolled into this study from January 2007 to December 2011. All cases were divided into DBE or SBE groups according to the endoscopic procedures they underwent. We then compared the diagnostic yield, the influence of DBE and SBE on the diagnostic/therapeutic course, the examination time, and post-procedure discomfort between DBE and SBE groups. RESULTS We observed no notable adverse events during or after the examinations. Additionally, SBE displays a significantly higher diagnostic rate (62.0%) than DBE (35.6%) via the anal approach (P=0.0137), while there was no difference in positive diagnostic rate between DBE and SBE via the oral route. Remarkably, it takes significantly less time to perform SBE examinations (38.86±5.64 minutes) than DBE procedures (41.80±6.50 minutes) via the oral route (P=0.048), although the average examination time for DBE is close to that for SBE via the anal route (P=0.952). However, DBE and SBE are similar in terms of their impact on the diagnostic/therapeutic course and complication rate. CONCLUSIONS Both SBE and DBE are very safe procedures to perform and SBE is a preferred choice for the evaluation of small bowel diseases in terms of diagnostic rate via the anal route compared with DBE.

MicroRNA-185 suppresses growth and invasion of colon cancer cells through inhibition of the hypoxia­inducible factor-2α pathway in vitro and in vivo.

MicroRNAs (miRs) are small non­coding RNAs with regulatory roles, which are involved in a broad spectrum of physiological and pathological processes, including cancer development and progression. However, the function of miR­185 in the development of human colon cancer has not yet been investigated. In this study, the association between miR­185 expression and the clinicopathological characteristics of patients with colon cancer was analyzed using quantitative polymerase chain reaction (qPCR). Using a gain­of­function approach, the effects of miR­185 overexpression on the expression of hypoxia­inducible factor­2α (HIF­2α), proliferating cell nuclear antigen (PCNA) and matrix metallopeptidase­2 (MMP­2) were investigated in SW620 colon cancer cells using qPCR and western blotting. Functional analysis of cellular proliferative activities, by MTT assay, and invasive potential, by Transwell assay, was conducted on SW620 cells expressing low levels of miR­185. miR­185 was found to be significantly downregulated in cancer tissues compared with adjacent non­cancerous tissues, and was negatively correlated with lymph node metastasis of colon cancer (P<0.001). miR­185 overexpression in vitro impeded cellular proliferation and invasive potential with reduced expression of HIF­2α, PCNA and MMP­2 in SW620 cells transfected with an miR­185 mimic. In addition, the tumor volumes in SW620 subcutaneous nude mouse models treated with miR­185 were significantly smaller than those of the control group. In conclusion, these findings indicate that miR­185 as a tumor suppressor may affect the development of colon cancer cells via inhibition of HIF­2α signaling, suggesting that miR­185 may serve as a potential therapeutic target in cancer treatment.