Urinary exosomal microRNA-145-5p and microRNA-27a-3p act as noninvasive diagnostic biomarkers for diabetic kidney disease

BACKGROUND Diabetic kidney disease(DKD),characterized by increased urinary microalbumin levels and decreased renal function,is the primary cause of end-stage renal di-sease.Its pathological mechanisms are complicated and multifactorial;Therefore,sensitive and specific biomarkers are needed.Urinary exosome originate from diverse renal cells in nephron segments and partially mirror the pathological changes in the kidney.The microRNAs(miRNAs)in urinary exosome are remark-ably stable and highly tissue-specific for the kidney.METHODS Type 2 diabetic mellitus(T2DM)patients were recruited from the Second Hospital of Hebei Medical University and were divided into two groups:DM,diabetic pa-tients without albuminuria[urinary albumin to creatinine ratio(UACR)<30 mg/g]and DKD,diabetic patients with albuminuria(UACR≥30 mg/g).Healthy subjects were the normal control(NC)group.Urinary exosomal miR-145-5p,miR-27a-3p,and miR-29c-3p,were detected using real-time quantitative polymerase chain reaction.The correlation between exosomal miRNAs and the clinical in-dexes was evaluated.The diagnostic values of exosomal miR-145-5p and miR-27a-3p in DKD were determined using receiver operating characteristic(ROC)analysis.Biological functions of miR-145-5p were investigated by performing RESULTS Urinary exosomal expression of miR-145-5p and miR-27a-3p was more upregulated in the DKD group than in the DM group(miR-145-5p:4.54±1.45 vs 1.95±0.93,P<0.001;miR-27a-3p:2.33±0.79 vs 1.71±0.76,P<0.05)and the NC group(miR-145-5p:4.54±1.45 vs 1.55±0.83,P<0.001;miR-27a-3p:2.33±0.79 vs 1.10±0.51,P<0.001).The exosomal miR-145-5p and miR-27a-3p positively correlated with albuminuria and serum creatinine and negatively correlated with the estimated glomerular filtration rate.miR-27a-3p was also closely related to blood glucose,gly-cosylated hemoglobin A1c,and low-density lipoprotein cholesterol.ROC analysis revealed that miR-145-5p had a better area under the curve of 0.88[95%confidence interval(CI):0.784-0.985,P<0.0001]in diagnosing DKD than miR-27a-3p with 0.71(95%CI:0.547-0.871,P=0.0239).Bioinformatics analysis revealed that the target genes of miR-145-5p were located in the actin filament,cytoskeleton,and extracellular exosome and were involved in the pathological processes of DKD,including apoptosis,inflammation,and fibrosis.CONCLUSION Urinary exosomal miR-145-5p and miR-27a-3p may serve as novel noninvasive diagnostic biomarkers or promising therapeutic targets for DKD.

Current early diagnostic biomarkers of prostate cancer

Prostate cancer （PCa） has become to have the highest incidence and the second mortality rate in western countries, affecting men＇s health to a large extent. Although prostate-specific antigen （PSA） was discovered to help diagnose the cancer in an early stage for decades, its specificity is relative low, resulting in unnecessary biopsy for healthy people and over-treatment for patients. Thus, it is imperative to identify more and more effective biomarkers for early diagnosis of PCa in order to distinguish patients from healthy populations, which helps guide an early treatment to lower disease-related mortality by noninvasive or minimal invasive approaches. This review generally describes the current early diagnostic biomarkers of PCa in addition to PSA and summarizes the advantages and disadvantages of these biomarkers.

Identification of potential diagnostic and prognostic biomarkers for breast cancer based on gene expression omnibus

BACKGROUND Breast cancer is regarded as a highly malignant neoplasm in the female population,posing a significant risk to women’s overall well-being.The prevalence of breast cancer has been observed to rise in China,accompanied by an earlier age of onset when compared to Western countries.Breast cancer continues to be a prominent contributor to cancer-related mortality and morbidity among women,primarily due to its limited responsiveness to conventional treatment modalities.The diagnostic process is challenging due to the presence of non-specific clinical manifestations and the suboptimal precision of conventional diagnostic tests.There is a prevailing uncertainty regarding the most effective screening method and target populations,as well as the specificities and execution of screening programs.AIM To identify diagnostic and prognostic biomarkers for breast cancer.METHODS Overlapping differentially expressed genes were screened based on Gene Expression Omnibus(GSE36765,GSE10810,and GSE20086)and The Cancer Genome Atlas datasets.A protein-protein interaction network was applied to excavate the hub genes among these differentially expressed genes.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses,as well as gene set enrichment analyses,were conducted to examine the functions of these genes and their potential mechanisms in the development of breast cancer.For clarification of the diagnostic and prognostic roles of these genes,Kaplan–Mei-er and Cox proportional hazards analyses were conducted.RESULTS This study demonstrated that calreticulin,heat shock protein family B member 1,insulin-like growth Factor 1,interleukin-1 receptor 1,Krüppel-like factor 4,suppressor of cytokine signaling 3,and triosephosphate isomerase 1 are potential diagnostic biomarkers of breast cancer as well as potential treatment targets with clinical implications.CONCLUSION The screening of biomarkers is of guiding significance for the diagnosis and prognosis of the diseases.

Role of mi RNAs and their potential to be useful as diagnostic and prognostic biomarkers in gastric cancer

Alterations in epigenetic control of gene expression play an important role in many diseases, including gastric cancer. Many studies have identified a large number of upregulated oncogenic mi RNAs and downregulated tumour-suppressor mi RNAs in this type of cancer. In this review, we provide an overview of the role of mi RNAs, pointing to their potential to be useful as diagnostic and/or prognostic biomarkers in gastric cancer. Moreover, we discuss the influence of polymorphisms and epigenetic modifications on mi RNA activity.

Serum microRNA expression profiling revealing potential diagnostic biomarkers for lung adenocarcinoma

Background:Recent studies have demonstrated that microRNAs(miRNAs)in the blood circulation can serve as promising diagnostic markers for cancers.This four-stage study aimed at finding serum miRNAs as potential biomarkers for lung adenocarcinoma(LA)diagnosis.Methods:The study was carried out between 2016 and 2017.The Exiqon miRNA qPCR panel(3 LA vs.1 normal control[NC]pooled serum samples)was used for initial screening to acquire miRNA profiles.Thirty-five dysregulated miRNAs were further evaluated in the training(24 LA vs.24 NCs)and testing stages(110 LA vs.110 NCs)using quantitative real-time polymerase chain reaction assays.Results:Four serum miRNAs(miR-133a-3p,miR-584-5p,miR-10b-5p,and miR-221-3p)were significantly overexpressed in LA patients compared with NCs.The diagnostic value of the four-miRNA panel was validated by an external cohort(36 LA vs.36 NCs).The areas under the receiver operating characteristic curve of the four-miRNA panel in the training,testing,and external validation stages were 0.734,0.803,and 0.894 respectively.Meanwhile,the expression level of miR-221-3p was much higher in LA tumor samples than that in the adjacent normal tissues(19 LA vs.19 NCs).The expression level of miR-10b-5p was also elevated in the serum-derived exosomes samples(18 LA vs.18 NCs).The expression of miR-133a-3p,miR-584-5p,and miR-10b-5p was significantly elevated in LA patients with epidermal growth factor receptor mutation compared with NCs.Conclusion:The study established a four-miRNA signature in serum that could improve the diagnostic capability of LA.

Novel biomarkers for early detection of gastric cancer

Gastric cancer(GC)remains a leading cause of cancer-related death worldwide.Less than half of GC cases are diagnosed at an advanced stage due to its lack of early symptoms.GC is a heterogeneous disease associated with a number of genetic and somatic mutations.Early detection and effective monitoring of tumor progression are essential for reducing GC disease burden and mortality.The current widespread use of semi-invasive endoscopic methods and radiologic approaches has increased the number of treatable cancers:However,these approaches are invasive,costly,and time-consuming.Thus,novel molecular noninvasive tests that detect GC alterations seem to be more sensitive and specific compared to the current methods.Recent technological advances have enabled the detection of blood-based biomarkers that could be used as diagnostic indicators and for monitoring postsurgical minimal residual disease.These biomarkers include circulating DNA,RNA,extracellular vesicles,and proteins,and their clinical applications are currently being investigated.The identification of ideal diagnostic markers for GC that have high sensitivity and specificity would improve survival rates and contribute to the advancement of precision medicine.This review provides an overview of current topics regarding the novel,recently developed diagnostic markers for GC.

Blood microRNAs as potential diagnostic and prognostic markers in cerebral ischemic injury

MicroRNAs are a family of small, genome-encoded endogenous RNAs that are transcribed but are not translated into proteins. They serve essential roles in virtually every aspect of brain function, including neurogenesis, neural development, and cellular responses leading to changes in synaptic plasticity. They are also implicated in neurodegeneration and neurological disorders, in responses to hypoxia and ischemia, and in ischemic tolerance induced by ischemic preconditioning. In recent developments, miRNA expres- sion profiling has been examined in stroke, and these studies indicate that miRNAs have emerged as key mediators in ischemic stroke biology. Both increased and decreased miRNA levels may be needed either as prevention or treatment of stroke. Novel approaches are being developed to get miRNA related therapeu- tics into the brain across an intact blood-brain barrier, including chemical modification, use of targeting molecules and methods to disrupt the blood-brain barrier.

Blood microRNAs as potential diagnostic markers for hemorrhagic stroke

Proper medical treatment of a stroke victim relies on accurate and rapid differentiation between ischemic and hemorrhagic stroke,which in current practice is performed by computerized tomography（CT） or magnetic resonance imaging（MRI） scans.A panel of micro RNAs could be an extremely useful clinical tool for distinguishing between hemorrhagic and ischemic stroke.This review has shown that blood miRNA profile can distinguish hemorrhagic from ischemic stroke in patients and in experimental animal models.It also seems likely they can differentiate between intracerebral and subarachnoid hemorrhage stroke.The miRNA profile in cerebrospinal fluid could be a useful diagnostic tool for subarachnoid hemorrhagic stroke.Decreased or increased miRNA levels may be needed either as prevention or treatment of stroke.Administration in vivo of miR-130 a inhibitor or miRNA mimic（miR-367,miR-223） in an intracerebral hemorrhage animal model improved neurological outcomes.

Alterations in the human oral microbiome in cholangiocarcinoma

Dear Editor,Alterations in the human microbiome are closely related to various hepatobiliary diseases.Gut microbial dysbiosis has been found in patients with cholangiocarcinoma(CCA)[1].However,the characteristics of oral microbiome in patients with CCA have not been studied.

血清免疫球蛋白GN-糖基的高通量分析——一种消化道癌症的非侵入性生物标志物

免疫球蛋白G(Immunoglobulin G,IgG)的N-糖基化在炎症性疾病的发展中起着重要作用。本研究旨在评价IgG N-糖基在消化道癌症亚型中的诊断效能。从中国医学科学院肿瘤医院招募749名消化道癌症患者,包括食管癌(esophageal cancer,EC)、胃癌(gastric cancer,GC)、结直肠癌(colorectal cancer,CRC)和胰腺癌(pancreatic cancer,PC)患者。采用亲水交互高效液相色谱-超高效液相色谱(hydrophilic interaction liquid chromatography using ultra-performance liquid chromatography,HILIC-UPLC)分析血浆中IgG的N-糖基构成。采用Bio-Plex悬液芯片系统检测方法(Bio-Rad)进行炎症因子检测。采用典型相关分析(canonical correlation analysis,CCA)探索糖基和炎症因子之间的相关性。采用LASSO回归和logistic回归模型,基于检测到的糖基谱建立可用于区分胃肠癌症患者和健康人群诊断模型。与健康对照组相比,EC、GC、CRC和PC患者的唾液酸化和半乳糖基化水平降低,而二等分乙酰葡萄糖胺基化水平在消化道癌症患者中升高。此外,只有胰腺癌患者具有低水平的岩藻糖基化。消化道癌症组的IL-1β、IL-31和sCD40L水平均高于对照组。IgG N-糖基的组成与炎症因子相关(r=0.556)。基于糖基的模型表现出良好的诊断效能,EC、GC、CRC和PC的受试者工作特征曲线下面积(AUC)分别为0.972、0.871、0.867和0.907。这些研究结果表明,IgG N-糖基在调节消化道肿瘤的发病机制中发挥了重要作用。血清IgG N-糖基可以作为潜在的非侵入性辅助消化道癌症临床诊断的方法。

Development and Evaluation of a Promising Biomarker for Diagnosis of Latent and Active Tuberculosis Infection

Background:Diagnosing latent tuberculosis(TB)infection(LTBI)and active TB(ATB)is crucial for preventing disease progression and transmission.However,current diagnostic tests have limitations in terms of accuracy and sensitivity,making it challenging to diagnose these different infection states.Therefore,this study intends to develop a promising biomarker for LTBI and ATB diagnosis to overcome the limitations of the current diagnostic tests.Methods:We developed a novelmultiepitope-based diagnostic biomarker(MEBDB)fromLTBI region of differentiation antigens using bioinformatics and immunoinformatics.Immune responses induced byMEBDM were detected using enzyme-linked immunosorbent spot and cytometric bead assays.This study was conducted from April 2022 to December 2022 in the SeniorDepartment of Tuberculosis at the 8thMedical Center of PLA General Hospital,China.Blood samples were collected from participants with ATB,individuals with LTBI,and healthy controls(HCs).The diagnostic efficacy of MEBDB was evaluated using receiver operating characteristic curves.Results:A novel MEBDB,designated as CP19128P,was generated.CP19128P comprises 19 helper T lymphocyte epitopes,12 cytotoxic T lymphocyte epitopes,and 8 B-cell epitopes.In silico simulations demonstrated that CP19128P possesses strong affinity for Toll-like receptors and elicits robust innate and adaptive immune responses.CP19128P generated significantly higher levels of tumor necrosis factor(TNF-α),interleukin 4(IL-4),and IL-10 in ATB patients(n=7)and LTBI(n=8)individuals compared with HCs(n=62)(P<0.001).Moreover,CP19128P-induced specific cytokines could be used to discriminate LTBI and ATB from healthy subjects with high sensitivity and specificity.Combining IL-2 with IL-4 or TNF-α could differentiate LTBI from HCs(the area under the receiver operating characteristic curve[AUC],0.976[95% confidence interval[CI],0.934-1.000]or 0.986[0.956-1.000]),whereas combining IL-4 with IL-17A or TNF-α could differentiate ATB from HCs(AUC,0.887[0.782-0.993]or 0.984[0.958-1.000]).Conclusions:Our study revealed that CP19128P is a potential MEBDBfor the diagnosis of LTBI andATB.Our findings suggest a promising strategy for developing novel,accurate,and sensitive diagnostic biomarkers and identifying new targets for TB diagnosis and management.

混配农药中毒防治研究

首次在我国开展大规模的混配农药中毒流行病学调查,发现接触混配农药是当前发生农药中毒的重要危险因素,为针对性的预防提供了科学依据。毒理学研究证实多数农药混剂呈增毒效应或协同作用,且与其毒代动力学变化有密切关系,为今后开发高效、低毒的混配农药提供了毒理学依据。在国内外首次建立甲基对硫磷单克隆抗体酶联免疫测定法,并成功研制便携式血胆碱酯酶快速测定仪;这些检测接触、效应生物标志物的灵敏、特异、便捷的方法已试用于现场,并具有开发为产品的前景。同时还发现对氧磷酶基因在192位点的单核苷酸多态性,可能是甲基对硫磷免疫毒性作用的易感性生物标志物之一。在总结不同类别农药混配后所致中毒的临床表现特点和规律的基础上,修订现行的农药中毒诊断标准,增添了有机磷单剂及混剂中毒所致“中间期肌无力综合征”的新内容,将提高我国农药中毒的诊治水平。本研究主要成果达国内领先和国际先进水平。

Lower and upper motor neuron involvement and their impact on disease prognosis in amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis is a fatal neurodegenerative disease characterized by progressive muscle wasting,breathing and swallowing difficulties resulting in patient’s death in two to five years after disease onset.In amyotrophic lateral sclerosis,both upper and lower motor neurons of the corticospinal tracts are involved in the process of neurodegeneration,accounting for great clinical heterogeneity of the disease.Clinical phenotype has great impact on the pattern and rate of amyotrophic lateral sclerosis progression and overall survival prognosis.Creating more homogenous patient groups in order to study the effects of drug agents on specific manifestations of the disease is a challenging issue in amyotrophic lateral sclerosis clinical trials.Since amyotrophic lateral sclerosis has low incidence rates,conduction of multicenter trials requires certain standardized approaches to disease diagnosis and staging.This review focuses on the current approaches in amyotrophic lateral sclerosis classification and staging system based on clinical examination and additional instrumental methods,highlighting the role of upper and lower motor neuron involvement in different phenotypes of the disease.We demonstrate that both clinical and instrumental findings can be useful in evaluating severity of upper motor neuron and lower motor neuron involvement and predicting the following course of the disease.Addressing disease heterogeneity in amyotrophic lateral sclerosis clinical trials could lead to study designs that will assess drug efficacy in specific patient groups,based on the disease pathophysiology and spatiotemporal pattern.Although clinical evaluation can be a sufficient screening method for dividing amyotrophic lateral sclerosis patients into clinical subgroups,we provide proof that instrumental studies could provide valuable insights in the disease pathology.

The significance of long non-coding RNAs in the pathogenesis,diagnosis and treatment of inflammatory bowel disease

Inflammatory bowel diseases(IBD)are a group of chronic relapsing gastrointestinal inflammatory diseases with significant global incidence.Although the pathomechanism of IBD has been extensively investigated,several aspects of its pathogenesis remain unclear.Long non-coding RNAs(lncRNAs)are transcripts with more than 200 nucleotides in length that have potential protein-coding functions.LncRNAs play important roles in biological processes such as epigenetic modification,transcriptional regulation and posttranscriptional regulation.In this review,we summarize recent advances in research on IBD-related lncRNAs from the perspective of the overall intestinal microenvironment,as well as their potential roles as immune regulators,diagnostic biomarkers and therapeutic targets or agents for IBD.

Biomarkers of resistance to immune checkpoint inhibitors in non-small-cell lung cancer:myth or reality?

Immune checkpoint inhibitors represent a major therapeutic advance in non-small-cell lung cancer with several approved anti-programmed death-1 and anti-programmed death-L1 immunotherapies.A majority of patients however,will not respond to immune checkpoint inhibitors and display primary resistance while a subset of initially responsive patients will present secondary resistance.Thus,there is a crucial need for biomarkers to enable better prediction and diagnosis,and to overcome such resistance.Along with improvement in the understanding of immune escape,new biomarkers are being developed,including large scale proteomic,genomic and transcriptomic approaches in tumor and blood samples.We review the novel biomarkers that have been investigated in non-small-cell lung cancer and discuss how they can rationalize therapeutic strategies.

Gut Microbiome in Colorectal Cancer:Clinical Diagnosis and Treatment

Colorectal cancer(CRC)is one of the most frequently diagnosed cancers and the leading cause of cancer-associated deaths.Epidemiological studies have shown that both genetic and environmental risk factors contribute to the development of CRC.Several metagenomic studies of CRC have identified gut dysbiosis as a fundamental risk factor in the evolution of colorectal malignancy.Although enormous efforts and substantial progresses have been made in understanding the relationship between human gut microbiome and CRC,the precise mechanisms involved remain elusive.Recent data have shown a direct causative role of the gut microbiome in DNA damage,inflammation,and drug resistance in CRC,suggesting that modulation of gut microbiome could act as a powerful tool in CRC prevention and therapy.Here,we provide an overview of the relationship between gut microbiome and CRC,and explore relevant mechanisms of colorectal tumorigenesis.We next highlight the potential of bacterial species as clinical biomarkers,as well as their roles in therapeutic response.Factors limiting the clinical translation of gut microbiome and strategies for resolving current challenges are further discussed.

Role of microRNAs in embryo–endometrial interactions:biological functions and clinical applications

The human endometrium and embryo both produce microRNAs(miRNAs),which are involved in various physiological activities.Accumulating evident suggested that endometrial miRNAs are regulated by steroid hormones that modulate endometrial functions,whereas embryo-derived miRNAs play vital roles in maternal–embryo communication.The aberrant expression of endometriumor embryo-derived miRNAs can cause early pregnancy and gestational disorders,including repeated implantation failure,recurrent miscarriage,and pathological conditions such as endometriosis.miRNAs are,therefore,ideal candidates for conducting clinicopathological tests.Potential diagnostic and prognostic tests using miRNAs are under development;however,the therapeutic applications of miRNAs in regulating gestational disorders require further investigation.

miRNA expression profiles in cerebrospinal fluid and blood of patients with Alzheimer’s disease and other types of dementia-an exploratory study

Background:MicroRNAs(miRNAs)are small non-coding RNA molecules that function as posttranscriptional regulators of gene expression.Measurements of miRNAs in cerebrospinal fluid(CSF)and blood have just started gaining attention as a novel diagnostic tool for various neurological conditions.The purpose of this exploratory investigation was to analyze the expression of miRNAs in CSF and blood of patients with Alzheimer’s disease(AD)and other neurodegenerative disorders in order to identify potential miRNA biomarker candidates able to separate AD from other types of dementia.Methods:CSF was collected by lumbar puncture performed on 10 patients diagnosed with AD and 10 patients diagnosed with either vascular dementia,frontotemporal dementia or dementia with Lewy bodies.Blood samples were taken immediately after.Total RNA was extracted from cell free fractions of CSF and plasma,and a screening for 372 known miRNA sequences was carried out by real time quantitative polymerase chain reactions(miRCURY LNA™Universal RT miRNA PCR,Polyadenylation and cDNA synthesis kit,Exiqon).Results:Fifty-two miRNAs were detected in CSF in at least nine out of ten patients in both groups.Among these,two miRNAs(let-7i-5p and miR-15a-5p)were found significantly up-regulated and one miRNA(miR-29c-3p)was found significantly down-regulated in patients with AD compared to controls.One hundred and sixty-eight miRNAs were frequently detected in the blood,among which miR-590-5p and miR-142-5p were significantly up-regulated and miR-194-5p was significantly down-regulated in AD patients compared to controls.Conclusions:Detection of miRNA expression profiles in blood and in particular CSF of patients diagnosed with different types of dementia is feasible and it seems that several expressional differences between AD and other dementia types do exist when measured in a clinically relevant setup.In this explorative pilot study,the deregulated miRNAs in CSF of AD patients may be associated with relevant target genes related to AD pathology,including APP and BACE1,which suggests that miRNAs are interesting candidates for AD biomarkers in the future.

Mice-lacking LMP2, immuno-proteasome subunit, as an animal model of spontaneous uterine leiomyosarcoma

Uterine tumors are the most common type of gynecologic neoplasm.Uterine leiomyosarcoma(LMS)is rare,accounting for 2%to 5%of tumors of the uterine body.Uterine LMS develops more often in the muscle tissue layer of the uterine body than in the uterine cervix.The development of gynecologic tumors is often correlated with female hormone secretion;however,the development of uterine LMS is not substantially correlated with hormonal conditions,and the risk factors are not yet known.Radiographic evaluation combined with PET/CT can be useless in the diagnosis and surveillance of uterine LMS.Importantly,a diagnostic biomarker,which distinguishes malignant LMS and benign tumor leiomyoma(LMA)is yet to be established.Accordingly,it is necessary to analyze risk factors associated with uterine LMS in order to establish a method of treatment.LMP2-deficient mice spontaneously develop uterine LMS,with a disease prevalence of~40%by 14 months of age.It is therefore of interest whether human uterine LMS shows a loss of LMP2 expression.We found LMP2 expression is absent in human LMS,but present in human LMA.Therefore,defective LMP2 expression may be one of the risk factors for LMS.LMP2 is potentially a diagnostic biomarker for uterine LMS,and gene therapy with LMP2-encording DNA may be a new therapeutic approach.

Recent advances in extracellular vesicles enriched with non-coding RNAs related to cancers

As membrane-bound structures that could be shedded by a parental cell,and fuse with others after shedding,and then release its contents,extracellular vesicles(EVs)are considered as an indispensable part of intercellular communication system.The EV contents might be all kinds of bioactive molecules including non-coding RNAs(ncRNAs),a large and complex group of RNAs with various subtypes that function to regulate biological events but classically do not code for proteins.In this review we covered the recently published works that validated the underlying molecular mechanisms regulating EV-associated ncRNAs’biogenesis,signaling,and particularly the systemic bio-effects related mostly to any stage of cancer progression,and the clinical potential of ncRNA-carrying EVs as diagnostic biomarkers and drugdelivery system that is being engineered for better loading and targeting capacity.Our views on the future direction of basic research and applications of EVs containing ncRNAs have also been shared.