Long expression of LY86-AS1 and HCG27_201 non-coding RNA in type 2 diabetes mellitus.

Long non-coding RNAs (LncRNAs) are non-coding RNAs. The potential roles of lncRNAs in type 2 diabetes mellitus (T2DM) are not well understood. In this study, our aim is to evaluate the expression levels of LY86-AS1 and HCG27_201 in patients with DM2 and a healthy control group. We obtained whole blood and serum samples from 100 T2DM and 100 non-diabetic subjects. Peripheral blood mononuclear cells (PBMC) were extracted from whole blood samples using Ficoll. Total RNA was isolated from PBMC obtained from type 2 diabetic patients and healthy control individuals using TRIzol LS reagent (GeneAll Biotechnology Co., LTD.). The extracted RNA was used for DNA synthesis (cDNA) with a reverse transcription kit (Takara). Real-time was performed with SYBR Green (Takara) and controlled by a Rotor-Gene system (Qiagen). We performed a quantitative PCR analysis of the lncRNA expression levels of LY86-AS1 and HCG27_201 in 200 samples. Here we found that the expression of LY86-AS1 and HCG27_201 was down-regulated in the T2DM group compared to the control group. We further determined that the expression of both lncRNAs was negatively correlated with serum glucose (FBS) levels. Receiver operating characteristic (ROC) analysis was used to evaluate the diagnostic value of LY86-AS1 and HCG27_201 as biomarkers of T2DM. ROC analysis showed that LY86-AS1 with a ROC area under the curve (AUC) 0.747 (P < 0.0001, sensitivity: 64.6 and specificity: 79.8) could be potential new prognostic biomarkers for DM2. The lower expression of the long-studied RNAs LY86-AS1 and HCG27_201 in patients with type 2 diabetes indicates their role in the pathogenesis of T2DM. Furthermore, LY86-AS1 can be used as a diagnostic marker for T2DM.

Downregulation of long non-coding RNAs LINC00523 and LINC00994 in type 2 diabetes in an Iranian cohort.

Long non-coding RNAs (lncRNAs) are a subclass within the repertoire of non-coding RNAs that have potential roles in type 2 diabetes mellitus (T2DM). However, the biological role and molecular mechanisms of lncRNAs in T2DM remain largely unknown. The purpose of this study is to investigate the relationship between LINC00523 and LINC00994 expressions and T2DM susceptibility in an Iranian cohort. In this case-control study, we obtained whole blood and serum samples from 100 T2DM patients and 100 healthy subjects. We extracted blood mononuclear cells (PBMC) from whole blood samples by Ficoll-Hypaque density gradient centrifugation. Total RNA was extracted from PBMC buffers using TRIzol-LS reagent (GeneAll). Finally, quantitative real-time PCR (qPCR) assay was used to detect the lncRNA expression levels of LINC00523 and LINC00994 in 200 samples. Expressions of LINC00523 and LINC00994 were significantly decreased in T2DM patients compared to healthy participants, with a change of 0.157 for LINC00523 and 0.159 for LINC00994. We observed a significant inverse relationship between the expressions of these lncRNAs with FBS. Receiver operating characteristic (ROC) curve analysis revealed that LINC00523 has a higher area under the ROC curve (AUC) of 0.7430 and a lower P-value (P<0.0001), as well as a sensitivity of 81.44% and a specificity of 61.11%. Therefore, LINC00523 can be considered as a potential prognostic biomarker for T2DM. It is likely that the lower expressions of the LncRNAs LINC00523 and LINC00994 in T2DM are related to the pathogenicity of T2DM in the Iranian population. Furthermore, LINC00523 can be considered as effective diagnostic biomarkers for T2DM.

Evaluation of DNA extraction methods and their clinical application for the direct detection of causal bacteria in culture fluids from continuous ambulatory peritoneal dialysis of patients with peritonitis by large-scale PCR.

The objectives of this study were to compare various DNA extraction methods and 16S rDNA primers and to assess the clinical utility of large-scale PCR in continuous peritoneal dialysis (CAPD) culture fluids.
Six strains of model organisms at dilutions of 10 (8) to 10 (0) colony-forming units (CFU)/mL were used to evaluate 5 DNA extraction methods and 5 primer-pair PCRs. Extensive PCR was applied to 100 CAPD culture fluids and the results were compared with those of conventional culture.
There were some differences between the different DNA extraction methods and primer sets with respect to detection limits. The InstaGene matrix (Bio-Rad Laboratories, USA) and the Exgene Clinic SV matrix (GeneAll Biotechnology Co. Ltd., Korea) seem to have higher sensitivity than others. Extended PCR results agreed with culture results in 97% of all cases (97/100). Two culture-positive cases that were PCR negative were identified as Candida albicans, and one culture-negative, PCR-positive specimen was identified as Bacillus circulans by sequencing. Two of the 54 positive broadband PCR products could not be sequenced.
There were differences in the analytical sensitivity of the various DNA extraction methods and primers for large-scale PCR. A broadband PCR assay for the detection of bacterial pathogens in CAPD culture fluid can be used as an adjunct to culture methods.


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Ying Yang 1 involved in brain pathology

Here we discuss data on the involvement of the transcription factor Yin Yang 1 (YY1) in the development of brain diseases, highlighting the mechanisms of its pathological actions. YY1 plays an important role in developmental and adult diseases of the nervous system. YY1 is essential for the maintenance and differentiation of neural and neuronal progenitors and oligodendrocytes that regulate neuronal and glial tissues in the brain. YY1 gene deficiency causes various anatomical and developmental abnormalities of the central nervous system (CNS). Once deregulated, YY1 exerts many neuropathological actions that are involved in the induction of several brain disorders such as stroke, epilepsy, Alzheimer’s disease, Parkinson’s disease, autism spectrum disorder, dystonia, and brain tumors. A better understanding of YY1 dysfunction in the nervous system may lead to the development of new therapeutic strategies related to the actions of YY1.
Keywords: YY1; Brain pathology gene regulation neurodegenerative disorders. Pediatric brain disorders.

Generation of CRISPR-Cas9-mediated epigenetic lines derived from human intestinal tissue by lentivirus transduction and single-cell cloning.

Human enteric tissue-derived intestinal tissues (HIEs; also called organelles) are a powerful ex vivo model for gastrointestinal research. Genetic modification of these non-transformed cultures allows new insights into gene function and biological processes associated with intestinal disease, as well as donor and gastrointestinal segment function. Here we present a detailed technical pipeline and protocol for using the CRISPR-Cas9 genome editing system to specifically delete a gene of interest in HIE via lentiviral transduction and single cell cloning. This protocol differs from a previously published variant that uses electroporation of the human colon to deliver transposons or CRISPR-Cas9 constructs, as this protocol uses LentiCRISPRv2 small guide RNAs modified and fused to express Cas9 and small guide RNAs in the lentivirus. . The protocol also includes steps for gene delivery and subsequent single-cell cloning, as well as transcriptional validation and sequencing of mutagenesis sites to create a knockout clone. An overview flowchart, step-by-step instructions, and troubleshooting tips are provided to help the investigator get on line for HIE removal in 2-3 months. In this protocol, we also describe how HIEs can be used as an ex vivo model to assess host restriction factors for viral replication (using human norovirus replication as an example) by removing host binding factors or immunity genes. . innate Other applications have been discussed to extend the utility of this system, e.g.

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