Diagnostic value of serum miR-141 in colorectal cancer and changes of serum miR-141 level after radical resection / 临床外科杂志

Objective To investigate the potential value of miR-141 as a diagnostic blood biomarker expression level in patients with colorectal cancer(CRC)and its change after radical CRC resection.Methods 75 CRC patients admitted to Affiliated to Medical College of Xi'an Jiaotong University 3201 Hospital from April 2019 to March 2021 were included in the experimental group,and 20 patients who received planned surgery for inguinal hernia during the same period were used as non-cancer control group.Surgical tissue and serum samples of these patients were collected.Microarray analysis was performed for miRNAs with significant expression differences in CRC tissues and serum.Real-time quantitative PCR(qRT-PCR)was used to verify the expression level of miR-141 in serum samples of the patients before surgery and at the 3rd day after surgery,and the relationship between miR-141 and clinicopathological characteristics of CRC patients was analyzed.Results By miRNA microarray analysis,we confirmed that 12 miRNAs were up-regulated simultaneously in tissue and serum samples of CRC patients,among which miR-141 was the most significantly up-regulated.Meanwhile,the relative expression level of miR-141 in serum of CRC group was significantly higher than that of non-cancer control group after qRT-PCR verification[2.50(1.06,3.12)vs.0.97(0.68,1.21),Z=-5.842,P＜0.05].According to ROC curve analysis,the AUC value of preoperative serum miR-141 for the diagnosis of CRC was 0.927(95％CI:0.862～0.992).When serum miR-141 ≥ 1.418,the accuracy of distinguishing between CRC and non-cancer control groups was 90.53％.Combined detection of serum miR-141 could increase the diagnostic AUC value of carcinoembryonic antigen and carbohydrate antigen-199 to 0.944(95％CI:0.899-0.998).For CRC patients,the relative expression level of serum miR-141 after radical resection was significantly lower than that before surgery[1.85(1.29,2.22)vs.2.55(2.07,3.18),Z=-5.416,P＜0.001].For those who did not receive radical resection,the relative expression level of serum miR-141 after surgery was slightly lower than that before surgery[2.28(1.72,2.74)vs.2.45(2.06,2.85)],but the difference was not statistically significant(Z=-1.917,P=0.055).The expression level of Mir-141 in serum of CRC patients was correlated with UICC stage and histological grade(P＜0.05).Conclusion Serum miR-141 reflects the pathological changes of CRC patients and can be used as a biomarker for non-invasive diagnosis of CRC.

Bioinformatics Analysis and Validation of Differential Expression of miRNAs in Plasma Exosomes from Patients with Active Rheumatoid Arthritis / 现代检验医学杂志

Objective To screen differentially expressed microRNAs(miRNAs)in plasma exosomes of active rheumatoid arthritis(RA)patients and healthy controls and conduct bioinformatics analysis for exploring the role and potential clinical application value of miRNAs in the pathogenesis of RA.Methods From January 2023 to April 2023,39 RA patients who visited the Rheumatology and Immunology Department of the Second Affiliated Hospital of Soochow University were selected as the study subjects,while 39 healthy individuals were selected as normal controls.The expression levels of miRNAs in plasma exosomes were detected by Illumina high-throughput sequencing technology,and the differentially expressed miRNAs were obtained by log2(Fold Change)absolute value>1 and P value<0.05.Six miRNAs were selected by the order from small to large P-value for bioinformatics analysis and validated using quantitative real-time fluorescence PCR(qRT-PCR).Results Compared with healthy controls,22 aberrantly expressed miRNAs were detected in plasma exosomes of RA patients,of which 4 were up-regulated and 18 were down-regulated.Among them,miR-30b-5p,miR-144-3p,miR-20a-5p,miR-223-5p,miR-425-3p,and miR-589-5p showed changed significantly.GO and KEGG enrichment analysis indicated that differentially expressed miRNAs may be involved in disease progression through regulation of signaling pathways such as TGF-β and PI3K/AKT,which were related to biological processes such as Th17 differentiation,intercellular interactions,and protein phosphorylation.The qRT-PCR validation results showed that the expression of miR-144-3p and miR-425-3p were significantly reduced in plasma exosomes of RA patients compared to healthy controls(t=3.617,3.595,all P<0.001),while the differences of miR-30b-5p,miR-223-5p,miR-589-5p,and miR-20a-5p expression were not statistically significant(t=1.956,1.331,1.662,1.861,all P>0.05).Conclusion The expression profile of plasma exosomal miRNAs changed in RA patients,which may be involved in disease progression through TGF-β and other signaling pathways.Exosome-derived miR-144-3p and miR-425-3p may be potential serological markers for RA diagnosis.

Si-Wu-Tang attenuates liver fibrosis via regulating lncRNA H19-dependent pathways involving cytoskeleton remodeling and ECM deposition / 中国天然药物

Liver fibrosis is a dynamic wound-healing response characterized by the agglutination of the extracellular matrix (ECM). Si-Wu-Tang (SWT), a traditional Chinese medicine (TCM) formula, is known for treating gynecological diseases and liver fibrosis. Our previous studies demonstrated that long non-coding RNA H19 (H19) was markedly upregulated in fibrotic livers while its deficiency markedly reversed fibrogenesis. However, the mechanisms by which SWT influences H19 remain unclear. Thus, we established a bile duct ligation (BDL)-induced liver fibrosis model to evaluate the hepatoprotective effects of SWT on various cells in the liver. Our results showed that SWT markedly improved ECM deposition and bile duct reactions in the liver. Notably, SWT relieved liver fibrosis by regulating the transcription of genes involved in the cytoskeleton remodeling, primarily in hepatic stellate cells (HSCs), and influencing cytoskeleton-related angiogenesis and hepatocellular injury. This modulation collectively led to reduced ECM deposition. Through extensive bioinformatics analyses, we determined that H19 acted as a miRNA sponge and mainly inhibited miR-200, miR-211, and let7b, thereby regulating the above cellular regulatory pathways. Meanwhile, SWT reversed H19-related miRNAs and signaling pathways, diminishing ECM deposition and liver fibrosis. However, these protective effects of SWT were diminished with the overexpression of H19 in vivo. In conclusion, our study elucidates the underlying mechanisms of SWT from the perspective of H19-related signal networks and proposes a potential SWT-based therapeutic strategy for the treatment of liver fibrosis.

MicroRNA-148b secreted by bovine oviductal extracellular vesicles enhance embryo quality through BPM/TGF-beta pathway

Background Extracellular vesicles (EVs) and their cargoes, including MicroRNAs (miRNAs) play a crucial role in cell-to-cell communication. We previously demonstrated the upregulation of bta-mir-148b in EVs from oviductal fluid of cyclic cows. This miRNA is linked to the TGF-β pathway in the cell proliferation. Our aim was to verify whether miR-148b is taken up by embryos through gymnosis, validate its target genes, and investigate the effect of miR-148b supplementation on early embryo development and quality. Methods Zygotes were cultured in SOF + 0.3% BSA (Control) or supplemented with: 1 μM miR-148b mimics during: D1-D7 (miR148b) or D1-D4 (miR148b-OV: representing miRNA effect in the oviduct) or D4-D7 (miR148b-UT: representing miRNA effect in the uterus) or 1 μM control mimics was used during: D1-D7 (CMimic). Embryos at ≥ 16-cells and D7 blastocysts (BD7) were collected to examine the mRNA abundance of transcripts linked to the TGF-β pathway (TGFBR2, SMAD1, SMAD2, SMAD3, SMAD5, BMPR2, RPS6KB1, POU5F1, NANOG), total cell number (TC), trophectoderm (TE), and inner cell mass (ICM) were also evaluated. One-way ANOVA was used for all analyses. Results We demonstrated that miR-148b can be taken up in both 16-cell embryos and BD7 by gymnosis, and we observed a decrease in SMAD5 mRNA, suggesting it's a potential target of miR-148b. Cleavage and blastocysts rates were not affected in any groups; however, supplementation of miR-148b mimics had a positive effect on TC, TE and ICM, with values of 136.4 ± 1.6, 92.5 ± 0.9, 43.9 ± 1.3 for miR148b and 135.3 ± 1.5, 92.6 ± 1.2, 42.7 ± 0.8, for miR148b-OV group. Furthermore, mRNA transcripts of SMAD1 and SMAD5 were decreased (P ≤ 0.001) in 16-cell embryos and BD7 from miR148b and miR148b-OV groups, while POU5F1 and NANOG were upregulated (P ≤ 0.001) in BD7 and TGFBR2 was only downregulated in 16-cell embryos. pSMAD1/5 levels were higher in the miR148b and miR148b-OV groups. Conclusions Our findings suggest that supplementation of bta-miR-148b mimics during the entire culture period (D1 - D7) or from D1 - D4 improves embryo quality and influences the TGF-β signaling pathway by altering the transcription of genes associated with cellular differentiation and proliferation. This highlights the importance of miR-148b on embryo quality and development.

miR-145-5p and miR-203a-5p overcome imatinib resistance in myelogenous leukemic cells through metabolic reprogramming-Supplemenntary data

Imatinib is the most effective therapy for chronic myeloid leukemia (CML), but many patients eventually develop resistance to it after an initial satisfactory response. This study investigated the potential of three miRNAs (miR-106b-5p, miR-145-5p, miR-203a-5p) in overcoming imatinib resistance in leukemic cells. The imatinib-resistant K562 (IR-K562) cells were developed and transfected with one of the three miRNAs to evaluate their potency in overcoming imatinib resistance. The changes in the metabolic profile were studied using flux balance analysis (FBA) and the data was validated using qRT-PCR.Among the three miRNAs, the ectopic expression of either miR-145-5p or miR-203a-5p was able to sensitize the IR-K562 cells to imatinib. The concentration of key oncometabolites; glucose, lactate, and glutamine, in the culture media of the miR-transfected IR-K562 cells, reverted to the same levels as seen in imatinib-sensitive K562 cells. In addition, the FBA analysis revealed that the metabolism of lipid, fatty acids, and electron transport chain were significantly altered in resistant cells. The FBA data was also validated at the molecular level. Interestingly, the imatinib treatment coupled with the transfection of miR-145-5p or miR-203a-5p cells could reverse the metabolic flux of IR-K562 to the levels seen in imatinib-sensitive K562 cells. This study highlights the key metabolic changes that occur during development of imatinib resistance. It also identifies the specific miRNAs which can be targeted to overcome imatinib resistance in CML.

miR-145-5p and miR-203a-5p overcome imatinib resistance in myelogenous leukemic cells through metabolic reprogramming

Imatinib is the most effective therapy for chronic myeloid leukemia (CML), but many patients eventually develop resistance to it after an initial satisfactory response. This study investigated the potential of three miRNAs (miR-106b-5p, miR-145-5p, miR-203a-5p) in overcoming imatinib resistance in leukemic cells. The imatinib-resistant K562 (IR-K562) cells were developed and transfected with one of the three miRNAs to evaluate their potency in overcoming imatinib resistance. The changes in the metabolic profile were studied using flux balance analysis (FBA) and the data was validated using qRT-PCR.Among the three miRNAs, the ectopic expression of either miR-145-5p or miR-203a-5p was able to sensitize the IR-K562 cells to imatinib. The concentration of key oncometabolites; glucose, lactate, and glutamine, in the culture media of the miR-transfected IR-K562 cells, reverted to the same levels as seen in imatinib-sensitive K562 cells. In addition, the FBA analysis revealed that the metabolism of lipid, fatty acids, and electron transport chain were significantly altered in resistant cells. The FBA data was also validated at the molecular level. Interestingly, the imatinib treatment coupled with the transfection of miR-145-5p or miR-203a-5p cells could reverse the metabolic flux of IR-K562 to the levels seen in imatinib-sensitive K562 cells. This study highlights the key metabolic changes that occur during development of imatinib resistance. It also identifies the specific miRNAs which can be targeted to overcome imatinib resistance in CML.

Mechanism of miRNA-related Signaling Pathway Against Diabetic Nephropathy and Intervention of Traditional Chinese Medicine： A Review / 中国实验方剂学杂志

Diabetic nephropathy （DN）， as one of the common chronic microvascular complications of diabetes， has become the main cause of renal failure in end-stage renal disease in China， increasing the risks of renal dialysis and kidney transplantation in diabetes patients. It is the leading cause of death in people with diabetes. The latest research on DN has focused on the gene level. microRNAs （miRNAs） are a family of endogenous accessible short-chain non-coding RNA molecules. By acting on a particular target， they activate or inhibit its mediated signaling pathways and related molecules， playing an important role in the occurrence and development of DN. They have become microeconomic factors for the prevention and treatment of DN. Traditional Chinese medicine （TCM） has a long history in the diagnosis and treatment of DN and has unique advantages such as significant curative effect and few side effects. A large number of studies have proved that TCM can target miRNA to affect multiple signaling pathways， participate in the regulation of inflammatory response， pyroptosis， mesenchymal transdifferentiation， and other pathological changes， and delay the further development of DN. Therefore， this study discusses the biogenesis mechanism of miRNA and its action mechanism in disease-related signaling pathways based on TCM diagnosis and treatment approaches from the perspective of miRNA， and summarizes the effect of TCM targeting miRNA on the disease-related signaling pathways and on DN. Thus， this study is expected to provide a theoretical reference for exploring the progress of TCM intervention in DN from the perspective of genes.

High-throughput screening of differential expression of exosomal miRNAs in DeBakey typeⅠacute aortic dissection patients / 中国胸心血管外科临床杂志

@#Objective    To evaluate the changes in the expression and significance of serum exosomal miRNAs in patients with DeBakey typeⅠacute aortic dissection (AAD). Methods    Twelve male patients with AAD and six healthy male medical examiners from our hospital were retrospectively included in this study. According to the time of chest pain, the AAD patients were divided into an AAD group within 24 h of chest pain onset, aged 47.00±8.79 years and an AAD group within 48 h of chest pain onset, aged 50.17±9.99 years. The healthy males were allocated to a control group, aged 49.17±4.26 years. Serum exosomal miRNAs were isolated, identified and quantified, and then differentially expressed exosomal miRNAs were screened. The bioinformatic analyses such as GO and KEGG were performed on the differentially expressed exosomal miRNAs. Results    High-throughput screening results revealed differential expression of AAD serum exosomal miRNAs. The upregulated miRNAs of AAD groups was hsa-miR-574-5p (P<0.05), and downregulated miRNAs were hsa-miR-223-3p, hsa-miR-146b-5p, hsa-miR-15b-5p, and hsa-miR-155-5p (P<0.05). Further bioinformatic analysis of the above miRNAs revealed that they were mainly enriched in signaling pathways such as transforming growth factor-β, cell cycle and endoplasmic reticulum protein synthesis. Conclusion    Differential expressions of serum exosomal miRNAs in AAD patients may be related to the pathogenesis of AAD, providing new ideas and clues for further exploration of AAD diagnostic markers and pathogenesis.

Exosomes secreted from human umbilical cord mesenchymal stem cells promote pancreatic ductal adenocarcinoma growth by transferring miRNAs / 中华肿瘤杂志

Objective: To observe the effects of exosomes derived from human umbilical cord mesenchymal stem cells on the proliferation and invasion of pancreatic cancer cells, and to analyze the contents of exosomes and explore the mechanisms affecting pancreatic cancer cells. Methods: Exosomes extracted from human umbilical cord mesenchymal stem cells were added to pancreatic cancer cells BxPC3, Panc-1 and mouse models of pancreatic cancer, respectively. The proliferative activity and invasion abilities of BxPC3 and Panc-1 cells were measured by cell counting kit-8 (CCK-8) and Transwell assays. The expressions of miRNAs in exosomes were detected by high-throughput sequencing. GO and KEGG were used to analyze the related functions and the main metabolic pathways of target genes with high expressions of miRNAs. Results: The results of CCK-8 cell proliferation assay showed that the absorbance of BxPC3 and Panc-1 cells in the hucMSCs-exo group was significantly higher than that in the control group [(4.68±0.09) vs. (3.68±0.01), P<0.05; (5.20±0.20) vs. (3.45±0.17), P<0.05]. Transwell test results showed that the number of invasion cells of BxPC3 and Panc-1 in hucMSCs-exo group was significantly higher than that in the control group (129.40±6.02) vs. (89.40±4.39), P<0.05; (134.40±7.02) vs. (97.00±6.08), P<0.05. In vivo experimental results showed that the tumor volume and weight in the exosomes derived from human umbilical cord mesenchymal stem cells (hucMSCs-exo) group were significantly greater than that in the control group [(884.57±59.70) mm(3) vs. (695.09±57.81) mm(3), P<0.05; (0.94±0.21) g vs. (0.60±0.13) g, P<0.05]. High-throughput sequencing results showed that miR-148a-3p, miR-100-5p, miR-143-3p, miR-21-5p and miR-92a-3p were highly expressed. GO and KEGG analysis showed that the target genes of these miRNAs were mainly involved in the regulation of glucosaldehylation, and the main metabolic pathways were ascorbic acid and aldehyde acid metabolism, which were closely related to the development of pancreatic cancer. Conclusion: Exosomes derived from human umbilical cord mesenchymal stem cells can promote the growth of pancreatic cancer cells and the mechanism is related to miRNAs that are highly expressed in exosomes.

Research progress of targeting miRNAs in prevention and treatment of ventricular remodeling after myocardial infarction / 中国药理学通报

Myocardial infarction is one of the leading causes of death worldwide, appearing at the last point of atherosclerotic plaque progression and unstable rupture. Impaired cellular signals after myocardial infarction leads to maladaptive changes, leading to ventricular remodeling and heart failure. MicroRNAs (miRNAs/miRs) are a kind of non-coding small RNAs that negatively regulate gene expression and regulate protein synthesis rate by changing the stability of targeted mRNA. This article reviews the latest research progress on the involvement of miRNAs in the progression of atherosclerotic plaques, the molecular mechanism of cardiac injury and subsequent remodeling during infarction, as well as the results of clinical studies, and puts forward the problems and limitations of targeted miRNAs in the treatment of cardiovascular diseases such as myocardial infarction and ventricular remodeling.

Peripheral origin exosomal microRNAs aggravate glymphatic system dysfunction in diabetic cognitive impairment

Cognitive dysfunction is one of the common central nervous systems (CNS) complications of diabetes mellitus, which seriously affects the quality of life of patients and results in a huge economic burden. The glymphatic system dysfunction mediated by aquaporin-4 (AQP4) loss or redistribution in perivascular astrocyte endfeet plays a crucial role in diabetes-induced cognitive impairment (DCI). However, the mechanism of AQP4 loss or redistribution in the diabetic states remains unclear. Accumulating evidence suggests that peripheral insulin resistance target tissues and CNS communication affect brain homeostasis and that exosomal miRNAs are key mediators. Glucose and lipid metabolism disorder is an important pathological feature of diabetes mellitus, and skeletal muscle, liver and adipose tissue are the key target insulin resistance organs. In this review, the changes in exosomal miRNAs induced by peripheral metabolism disorders in diabetes mellitus were systematically reviewed. We focused on exosomal miRNAs that could induce low AQP4 expression and redistribution in perivascular astrocyte endfeet, which could provide an interorgan communication pathway to illustrate the pathogenesis of DCI. Furthermore, the mechanisms of exosome secretion from peripheral insulin resistance target tissue and absorption to the CNS were summarized, which will be beneficial for proposing novel and feasible strategies to optimize DCI prevention and/or treatment in diabetic patients.

Could be FOXO3a, miR-96-5p and miR-182-5p useful for Brazilian women with luminal A and triple negative breast cancers prognosis and target therapy?

Abstract FOXO3a dysregulation is frequently implicated in tumorigenesis, and its inhibition can occur by several molecular mechanisms. Among these, post-transcriptional suppression by miRNAs has been associated with various cancers initiation. Here, we assessed the expression profiles of the most relevant miRNAs for breast tumorigenesis, using Luminal A (LA) and Triple-Negative (TN) breast cancer from Brazilian patients, by the quantitative real time-PCR method. Their potential prognostic role for the patients was also evaluated. We identified the miRNAs miR-96-5p and miR-182-5p, de-scribed as negative regulators of FOXO3A, with differential expression both in LA and TN tumors when compared to normal tissue. The miR-96-5p and miR-182-5p miRNAs were upregulated in LA (7.82 times, p < 0.005; 6.12 times, p < 0.005, respectively) and TN breast cancer samples (9.42 times, p < 0.0001; 8.51 times, p < 0.0001) compared to normal tissues. The samples with higher miR-96-5p and miR-182-5p expression (FR ≥ 4) were submitted for FOXO3a immunostaining. Reduced protein detection was observed in all of the tumors compared to normal tissues. The most prominent miRNA expression and FOXO3a protein suppression were observed in TN samples (p < 0.001), indicating the relevant role of these molecules in this tumor biology and clinical behavior. Our results corroborate the literature regarding to the relevance of FOXO3a in the breast cancer, and they open new perspectives for alternative target therapy options for Brazilian patients expressing both FOXO3a and its regulatory miRNAs.

microRNAS no líquido cefalorraquidiano associados à doença de Alzheimer: uma revisão sistemática e análise de vias usando a abordagem de aprendizado de máquina / microRNAs in Cerebrospinal Fluid Associated with Alzheimer's Disease: A systematic review and pathway analysis using a machine learning approach

O aumento da expectativa de vida e o envelhecimento populacional têm contribuído para o crescimento da prevalência da doença de Alzheimer (DA), uma vez que o envelhecimento é um dos fatores de risco para o desenvolvimento da forma esporádica da doença. O diagnóstico da DA é essencialmente baseado na avaliação clínica e requer a experiência de profissionais altamente treinados para um diagnóstico conclusivo. No entanto, devido à complexidade da doença e à necessidade de métodos mais precisos, a pesquisa por biomarcadores tem ganhado uma crescente importância. Nos últimos tempos, tem-se observado um aumento significativo no interesse pelo papel dos microRNAs (miRNAs) na regulação da expressão gênica e sua associação com a DA. Os miRNAs são pequenas moléculas de RNA não codificantes que desempenham um papel crucial na regulação de processos celulares. Vários miRNAs foram identificados como potenciais marcadores para o diagnóstico e progressão da DA. O presente estudo objetivou realizar uma busca por miRNAs diferencialmente expressos em líquor cefalorraquidiano (LCR) de pacientes com DA comparados a indivíduos cognitivamente saudáveis, a partir de banco de dados públicos e usando ferramentas de aprendizado de máquina, com validação dos resultados em uma revisão sistemática e a proposição de vias biológicas reguladas. Para isso, a primeira busca foi realizada na plataforma GEO Database e aplicado o algoritmo LightGBM. Posteriormente, a revisão sistemática foi realizada utilizando o PECO ­ população (P): indivíduos idosos, exposição (E): doença de Alzheimer (C): indivíduos cognitivamente saudáveis, desfeixo ou outcome (O): miRNAs diferencialmente expressos no líquor, usando os repositórios eletrônicos: MEDLINE/PubMed (Medical Literature Analysis and Retrieve System Online), Scopus, Cinahl, Web of Science e Embase. A análise de vias foi feita no miRTarBase. Após a sobreposição dos resultados obtidos pelo algoritmo e pela revisão sistemática, foram identificados sete miRNAs mais diferencialmente expressos no líquor de indivíduos com DA: miRNA-1274a, miRNA-193a-5p, miRNA-28-3p, miRNA-30a-3p, miRNA-145, miRNA-19b e miRNA-143. Na análise de enriquecimento de vias, foram identificadas: resposta ao dano no DNA por ATM, sinalização ERBB, sinalização de mensageiro secundário intracelular, sinalização MAPK e sinalização TGF-beta, as quais possuem participação na fisiopatologia da DA. Os resultados sugerem que os miRNA-1274a, miRNA-193a-5p, miRNA-28-3p, miRNA-30a-3p, miRNA-145, miRNA-19b e miRNA-143 podem estar envolvidos nos mecanismos moleculares e nas vias biológicas envolvidas na doença e podem ser no futuro alvos terapêuticos para a DA.

The increase in life expectancy and the aging population have contributed to the growth in the prevalence of Alzheimer's disease (AD), as aging is one of the risk factors for the development of the sporadic form of the disease. The diagnosis of AD is primarily based on clinical evaluation and requires the expertise of highly trained professionals for a conclusive diagnosis. However, due to the complexity of the disease and the need for more precise methods, research on biomarkers has gained increasing importance. In recent times, there has been a significant increase in interest in the role of microRNAs (miRNAs) in gene expression regulation and their association with AD. MiRNAs are small non-coding RNA molecules that play a crucial role in regulating cellular processes. Several miRNAs have been identified as potential markers for the diagnosis and progression of AD. This study aimed to search for differentially expressed miRNAs in the cerebrospinal fluid (CSF) of AD patients compared to cognitively healthy individuals using public databases and machine learning tools, with the validation of the results in a systematic review and the proposal of regulated biological pathways. To do this, the initial search was conducted on the GEO Database platform, and the LightGBM algorithm was applied. Subsequently, the systematic review was performed using the PECO framework ­ Population (P): elderly individuals, Exposure (E): Alzheimer's disease, Control (C): cognitively healthy individuals, Outcome (O): differentially expressed miRNAs in CSF, utilizing the electronic repositories: MEDLINE/PubMed, Scopus, Cinahl, Web of Science, and Embase. Pathway analysis was conducted using miRTarBase. After overlapping the results obtained by the algorithm and the systematic review, seven miRNAs were identified as the most differentially expressed in the CSF of individuals with AD: miRNA-1274a, miRNA-193a-5p, miRNA-28-3p, miRNA-30a-3p, miRNA-145, miRNA-19b, and miRNA-143. In the pathway enrichment analysis, the following pathways were identified: DNA damage response by ATM, ERBB signaling, intracellular second messenger signaling, MAPK signaling, and TGF-beta signaling, all of which have a role in the pathophysiology of AD. The results suggest that miRNA-1274a, miRNA-193a-5p, miRNA-28-3p, miRNA-30a-3p, miRNA-145, miRNA-19b, and miRNA-143 may be involved in the molecular mechanisms and biological pathways associated with the disease and could potentially serve as therapeutic targets for AD in the future.

Screening and verification of differentially expressed miRNAs in PBMCs of patients with silicosis / 中国职业医学

@#Abstract: Objective To screen and validate the differentially expressed microRNAs (miRNAs) in peripheral blood Methods - mononuclearcells(PBMCs)ofpatientswithsilicosis. Forty eightpatientswithoccupationalsilicosisatstageⅠ(case group)and45healthycontrols(controlgroup)wereselectedasresearchsubjectsbyrandomnumbertablemethod.PBMCswere-separatedbyFicollPaquegradientcentrifugationfromperipheralblood.Threepeoplefromeachgroupwererandomlyselected for miRNAs transcriptome sequencing. R Studio software was used to screen differentially expressed miRNAs, and FunRich software was used to predict the upstream transcription factors related to the differentially expressed miRNAs in PBMCs. The - Results differentially expressed miRNAs were verified by real time quantitative polymerase chain reaction. A total of 124 - - differentiallyexpressedmiRNAswerescreened,amongthem,97miRNAswereupregulatedand27miRNAswere down regulated.- The 67 targetgenes predicted by differentialmiRNAs were mainly involved in intracellularprocesses and nucleic acid binding transcriptionfactoractivities,includingcelladhesionmolecules,ratsarcoma,osteoclastdifferentiationandotherpathways.The-------------topfivemiRNAsdifferentiallyupregulatedwerehsamiR1373p,hsamiR500b3p,hsamiR190a5p,hsamiR1413pand------------hsamiR223p.ThetopfivedifferentiallydownregulatedmiRNAswerehsamiR2025p,hsamiR548ai,hsamiR55873p,------hsamiR5705pandhsamiR103983p.ThechangeofthesemiRNAswereconsistentwiththeresultspredictedaccordingto the miRNAs transcriptome sequencing. The transcription factors including specificity protein 1, early growth response 1, zinc Conclusion finger protein 161, etc., were obtained according to the differentially expressed miRNAs. The differentially

Construction of endogenous RNA regulatory network for colorectal cancer based on bioinformatics / 中南大学学报(医学版)

OBJECTIVES@#The high morbidity and mortality of colorectal cancer (CRC) have posed great threats to human health. Circular RNA (circRNA) and microRNA (miRNA), acting as competing endogenous RNAs (ceRNAs), have been found to play vital roles in carcinogenesis. This paper aims to construct a circRNA/miRNA/mRNA regulatory network so as to explore the molecular mechanism of CRC.@*METHODS@#The sequencing data of circRNA from CRC were obtained from Gene Expression Omnibus (GEO). The differential circRNA was screened and its structure was identified by Cancer-specific CircRNA Database (CSCD); the sequencing data of miRNA and messenger RNA (mRNAs) were downloaded from The Cancer Genome Atlas (TCGA) database and the differentially expressed genes were screened; the corresponding miRNA of differential circRNAs were predicted by CircInteractome database; DIANA, Miranda, PicTar, and TargetScan databases were used to predict the target genes of different miRNAs; the target genes from Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were enriched by R language; String database combined with Cytoscape 3.7.2 software was used to construct protein-protein interaction (PPI) network and hub genes were screened; the expressions of mRNAs in the Top10 hub genes were verified in CRC. The network diagrams of circRNAs/miRNAs/mRNAs and circRNAs/miRNAs/Top10 hub mRNAs were constructed by Cytoscape3.7.2. Real-time PCR was used to examine the expression levels of hsa_circRNA_0065173, hsa-mir-450b, hsa-mir-582, adenylate cyclase 5 (ADCY5), muscarinic acetylcholine receptor M2 (CHRM2), cannabinoid receptor 1 (CNR1), and lysophosphatidic acid receptor 1 (LPAR1) in the CRC tissues and the adjacent normal tissues.@*RESULTS@#A total of 14 differential circRNAs were identified, and 8 were found in CSCD; 34 miRNAs targeted by circRNAs were obtained. The PPI network was constructed, and the Top10 hub genes were identified, which were CHRM2, melanin concentrating hormone receptor 2 (MCHR2), G-protein gamma 3 subunit (GNG3), neuropeptide Y receptor Y1 (NPY1R), CNR1, LPAR1, ADCY5, adenylate cyclase 2 (ADCY2), gamma 7 (GNG7) and chemokine 12 (CXCL12), respectively. The expressions of Top 10 hub genes were also verified, and the results showed that the Top 10 hub genes were down-regulated in CRC; the constructed network diagram showed that hsa_circRNA_0065173 may regulate ADCY5, CHRM2, and Hsa-mir-450b by modulating hsa-mir-450b and hsa-mir-582. CNR1 and LPAR1 genes might serve as potentially relevant targets for the treatment of CRC. Real-time PCR results showed that the expression levels of hsa_circRNA_0065173, ADCY5, CHRM2, CNR1 and LPAR1 in the CRC tissues were significantly reduced compared with the adjacent normal tissues (all P<0.05); the expression levels of hsa-mir-450b and hsa-miR-582 were significantly increased (both P<0.05).@*CONCLUSIONS@#In this study, a potential circRNAs/miRNAs/mRNAs network is successfully constructed, which provides a new insight for CRC development mechanism through ceRNA mediated by circRNAs.

MiRNA expressions in peripheral blood plasma of workers in a certain mine / 中华放射医学与防护杂志

Objective:To investigate the effect of radon on the expressions of miR-16, miR-106b, miR-449a, let-7g, miR-21, miR-221 and miR-34a in peripheral blood plasma of miners.Methods:A total of 46 randomly selected miners worked underground(the underground group)and 38 miners worked aboveground (the control group). MiRNA levels in the underground and control groups were detected by qRT-PCR and their relationship with cumulative effective dose was further analyzed.Results:The levels of miR-106b, miR-21, miR-221 in plasma of the study group were significantly higher than those in the control group( Z=-2.32, -2.47, -2.79, P<0.05), the corresponding Fc values were 1.61, 1.75, 1.30, respectively. The levels of miR-16, miR-449a, let-7g and miR-34a were slightly higher than those in the control group ( P>0.05). After controlling of confounding factors such as age, BMI and smoking, the alteration of miR-16, miR-106b, let-7g, miR-21 and miR-221 in plasma of the underground group were positively correlated with the cumulative effective dose( t=2.50, 3.31, 2.60, 2.95, 3.25, P<0.05). No significant difference was observed in the plasma levels of miR-449a and miR-34a between the two groups ( P>0.05). Conclusions:miR-106b, miR-21 and miR-221 could be used as potential biomarkers of radon exposure.

Anti-inflammation effect of moxibustion for rats with diarrhea-predominant irritable bowel syndrome based on multiple miRNAs regulating NF-κB signal pathway / 中国针灸

OBJECTIVE@#To observe the effect of moxibustion on the regulation of nuclear factor-kappa B (NF-κB) and inflammatory factors by multiple microRNAs (miRNAs) in rats with diarrhea-predominant irritable bowel syndrome (IBS-D), and to explore the anti-inflammatory mechanism of moxibustion on IBS-D.@*METHODS@#Twelve of 52 newborn rats were randomly selected into a normal group. The remaining rats were made into IBS-D model. A total of 36 rats with successful model were randomly divided into a model group, a medication group and a moxibustion group, 12 rats in each group. The rats in the medication group were intraperitoneally injected with pyrrolidine dithiocarbamate (PDTC). The rats in the moxibustion group were treated with moxibustion at "Tianshu" (ST 25) and "Shangjuxu" (ST 37) for 20 min each time. All the intervention was given once a day for 7 days. Before and after modeling as well as after intervention, the body mass, loose stool rate and the minimum volume threshold of abdominal withdrawal reflex (AWR) were measured. After intervention, the contents of serum tumor necrosis factor α (TNF-α), interleukin (IL)-1β and IL-8 were detected by ELISA method; the morphology of colon tissues was observed by HE staining, and the expressions of miR-155, miR-125b, miR-29b, miR-31, miR-18a and NF-κB p65 mRNA in colon tissues were detected by real-time PCR. The expressions of NF-κB p65, TNF-α, IL-1β and IL-8 protein in colon tissues were detected by immunofluorescence.@*RESULTS@#After modeling, the body mass and the minimum volume threshold of AWR in the model group were lower than those in the normal group (P<0.01); the rates of loose stool in the model group were higher than those in the normal group (P<0.01); after intervention, in the model group, the inflammatory infiltration of colon tissues was obvious, and the serum levels of TNF-α, IL-1 β, IL-8 were higher than those in the normal group (P<0.05); the expression of miR-155, miR-125b, miR-29b, miR-31, miR-18a and NF-κB p65 mRNA in colon tissues was higher than that in the normal group (P<0.05); the protein expression of NF-κB p65, TNF-α, IL-1β, IL-8 was also higher than that in the normal group (P<0.01). After intervention, the body mass and the minimum volume threshold of AWR in the medication group and the moxibustion group were both higher than those in the model group (P<0.05); the loose stool rate in the medication group and the moxibustion group were lower than those in model group (P<0.05); the inflammatory cells infiltration in the colon tissues was less, the serum levels of TNF-α, IL-1β and IL-8 as well as the protein expression of NF-κB p65, TNF-α, IL-1β and IL-8 in the colon tissues in the medication group and the moxibustion group were lower than those in the model group (P<0.05, P<0.01). The expression of miR-125b, miR-31, miR-18a and NF-κB p65 mRNA in the medication group were lower than those in the model group (P<0.05). The expression of miR-155, miR-125b, miR-29b, miR-31, miR-18a and NF-κB p65 mRNA in the moxibustion group were lower than those in the model group (P<0.05). The miR-155, miR-125b, miR-29b, miR-31, miR-18a were positively correlated with NF-κB p65 mRNA (0<r<1, P<0.01).@*CONCLUSION@#The anti-inflammatory mechanism of moxibustion at "Tianshu" (ST 25) and "Shangjuxu" (ST 37) for IBS-D rats may be related to regulating multiple miRNAs to inhibit NF-κB signal pathway and reduce the expression of inflammatory factors.

Research progress of exosomal miRNA in skin wound healing and regeneration / 中国临床药理学与治疗学

Skin is the largest organ of the human body, and maintaining the integrity of the skin is very important to maintain the normal function of the body. Repair and regeneration after skin trauma is a complex dynamic process, which involves biological processes such as cell proliferation and migration, extracellular matrix synthesis and deposition, angiogenesis and remodeling. Recently, exocrine miRNA is increasingly considered as a potential therapy for the treatment of skin trauma. Exosomes, as intercellular messengers, can affect the function of target cells through fusion, endocytosis and receptor-ligand interaction. MiRNA, the main component of exosomes, plays an important role in all stages of wound repair. Therefore, understanding the specific role of exocrine miRNA in skin wound repair and regeneration may provide a powerful basis for the development of new treatments for skin trauma in the future. In this review, we summarized the role of exocrine miRNA in the three stages of inflammation, proliferation and remodeling of skin wound repair and regeneration, and expounded its mechanism. In addition, we also discussed the current limitations and future prospects of the study of exocrine miRNA in skin wound repair and regeneration, in order to provide ideas for follow-up research.

Avaliação do perfil de microRNAs exossomais em portadores de hipercolesterolemia familial / Evaluation of the profile of exosomal microRNAs in patients with familial hypercholesterolemia

A Hipercolesterolemia Familial (HF) é uma doença hereditária do metabolismo lipídico que causa aos portadores alta incidência de aterosclerose prematura. A HF pode ser diagnosticada clínica e geneticamente, entretanto, apenas cerca de 40% podem ter confirmados pelo diagnostico molecular. Assim, outros sistemas de diagnóstico devem ser avaliados. Ultimamente devido a estabilidade em fluidos biológicos, os exossomos circulantes apresentam grande potencial, pois carreiam um número variado de compostos e são considerados veículos de intercomunicação entre os tecidos. Sabe-se que vários RNAs são carreados nos exossomos, incluindo miRNAs, lncRNA e uma variedade de proteínas. Estes componentes podem ser marcadores de diagnóstico para várias doenças inclusive a HF e suas complicações cardiovasculares. Foram utilizadas amostras de exossomos plasmáticos provenientes de 54 pacientes HF sem uso de estatina por, no mínimo, seis semanas, e 38 indivíduos normolipidêmicos para sequenciamento de miRNAs e estudo da proteômica. Os exossomos foram isolados utilizando dois métodos precipitação química e cromatográfica de exclusão de tamanho e caraterizados utilizando: dispersão de luz dinâmica, Western blotting, rastreamento de nanopartículas (NanoSight), imunomarcação e microscopia eletrônica de transmissão. Os miRNAs e proteínas foram extraídos dos exossomos e analisados por sequenciamento de nova geração e espectrometria de massa, respectivamente. Os dados clínicos, biodemográficos e laboratoriais dos pacientes HF e controles indicaram diferenças significativas esperadas entre os grupos, indicando que foram selecionados adequadamente. A caracterização físico-química dos exossomos mostrou resultados com tamanho de ˜90nm e imunorreação positiva para tetraspaninas. O resultado do sequenciamento identificou acima 2000 miRNAs. Os miR-122- 5p e miR-21-5p apresentaram expressão aumentada no grupo HF (log2FC=1,79 e log2FC=1,27, respectivamente), e o miR-122-5p pós normalização em relação ao controle manteve significativo comparados ao controle (p=0,034). A análise comparativa entre exossomos e plasma total mostrou diferença significativa, pois foram identificadas 239 proteínas (p <0,05) diferentes entre exossomos e plasma. Em exossomos, 17 proteínas foram aumentadas e 21 diminuídas em pacientes com HF em comparação com o controle (p <0,05). Destas, seis proteínas foram mais abundantes em HF e sete proteínas foram menos abundantes em exossomos de pacientes com HF em comparação com o controle. A análise de enriquecimento por bioinformática mostrou que a maior parte dessas moléculas (miRNAs e proteínas) foram relacionadas com metabolismo lipídico, dislipidemia, aterosclerose, doença arterial coronariana, adipogênese. Assim, na busca de novos alvos como potenciais biomarcadores de diagnóstico da HF, nossos resultados da análise integrativa entre os miRNAs e as proteínas exossomais abre novas frentes de pesquisa mais bem direcionadas, para a validação desses miRNAs e proteínas exossomais

Familial Hypercholesterolemia (FH) is an inherited disease of lipid metabolism that causes a high incidence of premature atherosclerosis in patients. FH can be diagnosed clinically and genetically, however, only about 40% can be confirmed by molecular diagnosis. Thus, other diagnostic systems should be evaluated. Lately, due to stability in biological fluids, circulating exosomes have great potential, as they carry a varied number of compounds and are considered vehicles of intercommunication between tissues. Several RNAs are known to be carried on exosomes, including miRNAs, lncRNA, and a variety of proteins. These components can be diagnostic markers for several diseases including FH and its cardiovascular complications. Plasma exosome samples from 54 FH patients without statin use for at least six weeks and 38 normolipidemic individuals were used for miRNA sequencing and proteomics studies. Exosomes were isolated using two methods chemical precipitation and size exclusion chromatography and characterized using: dynamic light scattering, Western blotting, nanoparticle tracking (NanoSight), immunostaining and transmission electron microscopy. MiRNAs and proteins were extracted from exosomes and analyzed by next-generation sequencing and mass spectrometry, respectively. Clinical, biodemographic and laboratory data of FH patients and controls indicated significant expected differences between the groups, indicating that they were appropriately selected. The physicochemical characterization of exosomes showed results with a size of ˜90nm and positive immunoreaction for tetraspanins. The sequencing result identified above 2000 miRNAs. miR-122-5p and miR-21-5p showed increased expression in the FH group (log2FC=1.79 and log2FC=1.27, respectively), and miR122-5p after normalization in relation to the control remained significant compared to the control (p=0.034). The comparative analysis between exosomes and total plasma showed a significant difference, as 239 different proteins (p < 0.05) were identified between exosome and plasma. In exosomes, 17 proteins were increased and 21 decreased in FH patients compared to control (p < 0.05). Of these, six proteins were more abundant in FH and seven proteins were less abundant in exosomes from patients with FH compared to the control. Bioinformatics enrichment analysis showed that most of these molecules (miRNAs and proteins) were related to lipid metabolism, dyslipidemia, atherosclerosis, coronary artery disease, adipogenesis. Thus, in the search for new targets as potential diagnostic biomarkers of FH, our results of the integrative analysis between miRNAs and exosomal proteins opens new and better-directed research fronts for the validation of these miRNAs and exosomal proteins

Research progress in miRNAs mediated radiation-induced by stander effect / 中华放射医学与防护杂志

Radiation-induced bystander effect (RIBE) refers to that irradiated cells release signaling factors and induce responses in nonirradiated cells.In other words, it is the communication between irradiated and nonirradiated cells by intracellular signals. RIBE could influence the efficacy of tumor radiotherapy, but also has potential risk to the normal tissues outside of radiation field. Studies have found that ionizing radiation can induce the alteration of miRNA expression not only in the irradiated cells but also in adjacent nonirradiated tissues, and miRNAs may play an important role in the regulation of signaling pathways between irradiated and nonirradiated bystander cells. This article reviewed the roles of miRNAs in RIBE.