Recently, the incidence and mortality of heat stroke (HS) have risen catastrophically. However, postmortem diagnosis of HS is challenging owing to the lack of characteristic morphological markers. Liver damage can often serve as a direct cause of death in HS. Therefore, this study aimed to identify protein biomarkers in the liver and to evaluate their utility as diagnostic biomarkers for HS. The morphological and biochemical tests of HS rats models revealed hydropic degeneration, hepatocyte necrosis, and impaired liver function. 283 differentially expressed proteins between the HS and control groups were screened by proteomic analysis. Subsequently, HSPA4, GGCX, and CYP2B6 were selected as candidate biomarkers based on Proteomic results, Western blotting and immunohistochemistry results in HS rats. These three candidate biomarkers were further validated as diagnostic protein biomarkers for HS death based on the immunohistochemistry results of 29 human cases. Finally, receiver operating characteristic analysis indicated that the combination of HSPA4, GGCX and CYP2B6 provided optimal diagnostic efficacy for HS, with an area under the curve of over 0.999. In conclusion, we propose that the integrating morphological findings, liver function analysis, and protein biomarkers (HSPA4, GGCX, and CYP2B6) in liver could be used for HS diagnosis in forensic practices.

We report for the first time the phenomenon of continuously color-tunable ECL from individual Au NCs confined in a porous hydrogel matrix. The color-tunable ECL property arises from the dynamic surface reconstruction of Au NCs and co-reactants.

Quantum dots (QDs) have become promising electrochemiluminescence (ECL) emitters with high quantum yield and size-tunable luminescence. However, most QDs generate strong ECL emission at the cathode, developing anodic ECL-emitting QDs with excellent performance is challenging. In this work, low-toxic quaternary AgInZnS QDs synthesized by a one-step aqueous phase method were used as novel anodic ECL emitters. AgInZnS QDs exhibited strong and stable ECL emission and a low excitation potential, which could avoid the side reaction of oxygen evolution. Furthermore, AgInZnS QDs displayed high ECL efficiency (ΦECL) of 5.84, taking the ΦECL of Ru(bpy)32+/tripropylamine (TPrA) ECL system as 1. Compared to AgInS2 QDs without Zn doping and traditional anode luminescent CdTe QDs, the ECL intensity of AgInZnS QDs was 1.62 times and 3.64 times higher than that of AgInS2 QDs and CdTe QDs, respectively. As a proof-of-concept, we further designed an “on-off-on” ECL biosensor for detecting microRNA-141 based on a dual isothermal enzyme-free strand displacement reaction (SDR), which not only to achieve the cyclic amplification of the target and ECL signal, but also to construct a switch of the biosensor. The ECL biosensor had a wide linear range from 100 aM to 10 nM with a low detection limit of 33.3 aM. Together, the constructed ECL sensing platform is a promising tool for rapid and accurate diagnosis of clinical diseases.

The recognition efficiency of walking strands and track strands is one of the vital factors to ensure the walking rate of DNA walkers. Herein, a skillful strategy was proposed to enhance the recognition efficiency via the design of DNA ring-supported quadruped DNA walker and rigid T-type quenching probes (T-QPs) tracks, which achieved the bio-stability and walking continuity for highly sensitive determination of Dam methyltransferase (Dam MTase) activity. Notably, DNA ring performed as DNA scaffold to reduce the folding and entanglement of the walking strands, and the T-QPs with ordered and controllable spatial morphology were immobilized on solid electrodes through high affinity between poly-cytosine (poly-C) of T-QPs and silver nanoparticles (Ag NPs), which improved identification efficiency of walking strands on orbits. As a proof of concept, the walking rate of proposed quadruped DNA walker was about 2-fold faster than that of unipedal DNA walker under the same conditions. Based on the combination of DNA ring-supported quadruped DNA walker and the ternary electrochemiluminescence (ECL) system of zirconium-based metal-organic framework/tripropylamine/Ag NPs (ZrMOF/TPrA/Ag NPs), the sensitive ECL sensing platform was constructed to monitor the Dam MTase activity with the low limit of detection of 6.25 x 10-7 U/mL. This work provides an approach for rational structural design of DNA walkers and develops a novel route for DNA-anchored solid electrodes.

5-Methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) are two of the most abundant epigenetic marks in mammalian genomes, and it has been proven that these dual epigenetic marks give a more accurate prediction of recurrence and survival in cancer than the individual mark. However, due to the similar structure and low expression of 5mC and 5hmC, it is challenging to distinguish and quantify the two methylation modifications. Herein, we employed the ten-eleven translocation family dioxygenases (TET) to convert 5mC to 5hmC via a specific labeling process, which realized the identification of the two marks based on a nanoconfined electrochemiluminescence (ECL) platform combined with the amplification strategy of a recombinase polymerase amplification (RPA)-assisted CRISPR/Cas13a system. Benefiting from the TET-mediated conversion strategy, a highly consistent labeling pathway was developed for identifying dual epigenetic marks on random sequence, which reduced the system error effectively. The ECL platform was established via preparing a carbonized polymer dot embedded SiO2 nanonetwork (CPDs@SiO2), which exhibited higher ECL efficiencies and more stable ECL performance compared to those of the scattered emitters due to the nanoconfinement-enhanced ECL effect. The proposed bioanalysis strategy could be employed for the identification and quantification of 5mC and 5hmC in the range from 100 aM to 100 pM, respectively, which provides a promising tool for early diagnosis of diseases associated with abnormal methylation.

In spite of the DNA walkers executing the signal accumulation task in the process of moving along the predetermined paths, the enhancement of walking dynamics and walking path controllability are still challenging due to the unprogrammed arrangements of DNA orbits. Taking these dilemmas into account, a bipedal DNA walker was designed skillfully by the virtue of wireframe orbits assembled by DNA cubes in order, which improved the efficiency and the continuity of walking. It could be attributed to the fact that both the contact chance and the dynamic interaction between walking strands and designated orbits were beneficial to minimize the possibility of derailment and improve the accumulation of signal. In addition, the hollow titanium dioxide nanospheres coated with rubrene (Rub@TiO2 NSs) were prepared by the etching of inner silicon dioxide nanoparticles (SiO2 NPs) to regulate the distribution pattern of rubrene (Rub) molecules and expose more electrochemically active sites for high-efficient electrochemiluminescence (ECL). Benefiting by the pore confinement-enhanced ECL, the electron and mass transfer was significantly accelerated because of the hollow structure of Rub@TiO2 NSs. Subsequently, endogenous dissolved oxygen as the coreactant and palladium nanoparticles (Pd NPs) as the coreaction accelerator were employed to constitute a ternary ECL system with explosive signal response. Combining with this ECL platform, the bipedal walker activated by the target can autonomously and directionally move on the DNA wireframe orbits to release the quenching probes continuously. In this way, the biosensor displayed a low detection limit (2.30 × 10-8 U·mL-1) and a wide linear range (1.0 × 10-7 to 1.0 × 10-1 U·mL-1) for the sensitive detection of Dam methyltransferase (Dam MTase) activity. Therefore, a novel strategy for the accurate quantification of epigenetic targets was developed by virtue of improving the walking dynamics of DNA walker and amplifying the ECL of Rub molecules.

Despite the intrinsic programmability and flexible biological functions of DNA walkers, it remains challenging for traditional DNA walkers to precisely control mechanical running paths in nanoscale spaces. In this work, an elegant dual engine-triggered DNA walker was elaborated based on precise orientation protocol, applying the DNA cube as the scaffold for anchoring the dual swing arms as engines and tracks as signal switches. The order arrangement and dual engines enhanced the operating efficiency and controllability of the DNA walker markedly. Moreover, the electrochemiluminescence (ECL) molecule perylene doped-titanium dioxide (Pe-TiO2) organic-inorganic nanospheres with low cost and high reproducibility were synthesized as the ECL emitters by one-step sol-gel method. It was worth noting that TiO2 gel not only restricted the stacking mode of Pe molecules in vibration and rotation, but also enhanced the interfacial charge transfer ability in organic-inorganic hybrid mode, resulting in the improvement of ECL efficiency and structural stability. Based on the luminous Pe-TiO2 nanospheres, a ternary ECL system was proposed with S2O82- as coreactant, and silver nanoparticles (Ag NPs) as coreaction accelerator. Ultimately, by combing the dual engine-triggered DNA walker with the ternary ECL system, an efficient ECL bioassay platform was developed for microRNA let 7a (let-7a) detection with a wide linear range from 10 fmol/L to 100 nmol/L and a low detection limit of 7.0 fmol/L. Hence, this work offered a universal strategy to predesign walking paths and improve loading capacity in virtue of DNA nanostructures, as well as to expand the biology research of planar organic luminophores in an organic-inorganic hybrid manner.

Despite the various synthesis approachs to obtain luminous carbon dots(CDs),it is still quite challenging to construct the efficient electrochemiluminescence(ECL)owing to their low ECL reactivity and easy ag-glomeration.Herein,an efficient and concise ECL system was skillfully constructed by taking advantage of the nitrogen and sulfur co-doped CDs(N,S-CDs)with surfaces rich in hydrazide groups as luminophors to emit intense ECL,and metal-organic framework(MOF)as the matrix to confine CDs in its nanospace.Sur-prisingly,the proposed CDs assembled MOF(CDs/ZIF-8)enhanced anodic ECL signal up to 250％of pure CDs under the exogenous coreactant-free condition.As a proof of concept,the highly sensitive detection of uric acid(UA)was realized by the constructed ECL platform with a low detection limit of 3.52 nmol/L ranging from 10 nmol/L to 50 μmol/L.This work expanded ideas for the application of pore confinement effect,and provided references for the detection of disease biomarkers of gout and hyperuricemia.

目的 探讨肝病患者血清抗核抗体谱(ANAs)分布情况.方法 回顾性分析2020年8月至2021年4月在首都医科大学附属北京地坛医院就诊的2148例门诊或住院的肝病患者,应用免疫印迹法检测血清ANAs.结果 2148例肝病患者中,ANAs阳性683例,阳性率31.80％,特异性自身抗体阳性率>10％的抗体有抗Ro-52抗体(25％)、抗AMA-M2抗体(14％)、抗SSA抗体(11％).不同病因所致肝脏疾病均可出现ANAs阳性,免疫性、药物性、混杂因素、其他、病毒性、酒精性、脂肪性肝病和不明原因肝功能异常组ANAs阳性率依次为74.87％、36.37％、36.02％、30.36％、27.63％、27.21％、25.56％和20.67％.自身免疫性肝病组ANAs阳性率(74.87％)高于其余病因组,不明原因肝功能异常组ANAs阳性率(20.67％)低于免疫性、药物性、混杂因素、其他、病毒性肝病组,差异均具有统计学意义(P<0.05).药物性、混杂因素、其他肝病与病毒性肝病组阳性率差异无统计学意义(P>0.05).酒精性肝病、脂肪肝与不明原因肝功能异常阳性率差异无统计学意义(P>0.05).自身免疫性肝病抗SSA抗体(16.23％)、抗SSB抗体(8.90％)、抗CENP-B抗体(20.94％)、抗Ro-52抗体(37.70％)、抗AMA-M2抗体(47.12％)阳性率高于其余病因组,差异具有统计学意义(P<0.01).结论 抗核抗体谱检测对不同病因肝病患者的诊断和鉴别诊断具有一定价值.

Glioma is the most frequent primary malignant brain tumor, which is characterized by high incidence and mortality, with a poor prognosis. Numerous studies have revealed the abnormal expression of long non-coding RNAs in gliomas. This study explored the effects and potential mechanism of LINC00663 in glioma. The LINC00663 levels and their prognostic values were analyzed from the GEO databases using bioinformatics. Also, LINC00663 expression in tissue samples and cell lines was measured using qRT-PCR. The roles of LINC00663 in glioma were confirmed using CCK8, EdU assay as well as Transwell tests. Moreover, the influences of LINC00663 on the AKT/mTOR signal cascades were detected using western blotting assay. LINC00663 expression was higher in both glioma tissues and cell lines than that in the normal brain tissues and human astrocytes. High expression of LINC00663 led to the low overall survival rate of patients with glioma. LINC00663 knockdown notably restrained cell proliferation, migration, and invasion abilities by decreasing the activation of AKT and mTOR. This study indicated that LINC00663 might have a cancer-promoting role in accelerating glioma development and progression through regulating AKT/mTOR pathway.

Here, we described a novel swing arm location-controllable DNA walker based on the DNA tetrahedral nanostructures (DTNs) for nucleic acid detection using the polycyclic aromatic hydrocarbon (PAH) microcrystals (TAPE-Pe MCs) consisting of the nonplanar molecular tetrakis(4-aminophenyl)ethene (TAPE) and planar molecular perylene (Pe) as electrochemiluminescence (ECL) luminophores. Specifically, the swing arm strands and track strands were fixed simultaneously on the DTNs to obtain the location-controllable DNA walker, which possessed an improved reaction efficiency compared to that of a fixed swing arm-based DNA walker due to the quantitative and orderly swing arm on the DTNs. On the other hand, the Pe microcrystals doped by TAPE molecules could decrease the π-π stacking of Pe molecules for the ECL efficiency enhancement, achieving a blue-shifted and intense ECL emission. Therefore, we defined this enhanced and blue-shifted ECL phenomenon as "inhibition of conjugation-driven ECL (IC-ECL)". To prove these principles, a location-controllable DNA walker-based ECL biosensor was developed with microRNA let-7a as target molecules. The ECL biosensor achieved a low detection limit of 4.92 fM within a wide linear range from 10 fM to 100 nM. This approach offers a new insight for ECL efficiency increase and location-controllable strategies with improved reaction efficiency, demonstrating potential in diagnostic analysis.

A highly sensitive L-Cys (L-cysteine) biosensor was constructed via silver nanoclusters loaded on the surface of silica nanoparticles (Ag NCs-SiO2 NPs) as an electrochemiluminescence (ECL) nanocomposite modified on the surface of glassy carbon electrode. The surface of silica nanoparticles (SiO2 NPs) was functionalized with amino and sulfhydryl groups by bovine serum protein (BSA) , and then silver nanoclusters (Ag NCs) were in situ reduced on the surface of SiO2 NPs by using BSA as a template and a reducing agent, which realized the stable load of silver nanoclusters. The Ag NCs-SiO, NPs electrochemiluminescence nanocomposites with the enhanced cathodic ECL signal were successfully obtained. With the existence of L-Cys in the solution, it covalently binds to Ag NCs forming Ag-S bond and quenching the ECL signal. Based on the above principle, a biosensor with the on-off signal response mode is constructed for the detection of L-Cys. The concentration range of L-Cys detected by the biosensor is 50 nmol/L-50 mu mol/L. At the same time, the detection limit is 13.7 nmol/L, which can realize the sensitivity and specificity analysis of L-Cys. The biosensor constructed above is expected to be extensively applied in biological , medical and other fields.

Objective:To explore the clinical value of autoantibodies in patients with liver disease.Methods:We retrospectively analyzed the data of 1 495 outpatients or inpatients with liver disease in Beijing Ditan Hospital of Capital Medical University from August 2020 to April 2021. Indirect immunofluorescence and Western blot were used to detect antinuclear antibody (ANA) and antinuclear antibodies (ANAs).Results:ANA and ANAs were positive in patients with liver diseases of various etiologies. Among 1 495 patients with liver disease, 494 cases were ANA positive, the positive rate was 33.04%; 573 cases were positive for ANAs, the positive rate was 38.33%. The positive rate of ANA in the immune liver disease group (63.37%) was higher than that in the viral, alcoholic, fatty liver, confounding factors and other liver disease groups, and the difference was statistically significant ( P<0.01). The ANA positive rate between the viral, alcoholic, fatty liver, and confounding factor groups was statistically significant ( χ2=19.823, P<0.01), the positive rate of ANAs in the immune liver disease group (80.23%) was higher than that in other liver disease groups, and the difference was statistically significant ( P<0.05). The antibody titer of immune liver disease group was mainly 1∶1000, and other liver disease etiology groups was mainly 1∶100. The two most common fluorescent karyotypes in liver disease groups of different etiologies are cytoplasmic and nuclear granular types. The most common specific antibody in the immune liver disease group was anti-mitochondrial antibody type 2 (anti-AMA-M2) antibody, the most common anti-Ro-52 antibody in viral, drug-induced, complex etiology, and other etiological groups, and the most common anti-SSA antibody in alcoholic liver disease. Anti-SSA antibody (17.44%), anti-SSB antibody (9.30%), anti-CENP-B antibody (22.09%), anti-Ro-52 antibody (41.28%), anti-AMA-M2 antibody (51.74%) were positive in immune liver disease group, The rate was higher than that of other liver disease etiology groups, and the difference was statistically significant ( P<0.01). When the ANA fluorescence karyotype is nuclear granule type, the positive rate of anti-CENP-B antibody, anti-Ro-52 antibody, and anti-AMA-M2 antibody in the immune liver disease group was higher than that in the viral liver disease group ( P<0.01), The positive rate of anti-Ro-52 antibody was higher than that of drug-induced liver disease group ( P<0.05). Conclusions:The ANA titer of autoimmune liver disease was mainly (1∶1 000). ANAs were mainly positive for anti-SSA antibody, anti-SSB antibody, anti-CENP-B antibody, anti-Ro-52 antibody, and anti-AMA-M2 antibody, especially anti-AMA-M2 antibody. When combined with ANA fluorescent karyotype and ANAs for analysis, if the fluorescent karyotype is nuclear particle type, the positive anti-Ro-52 antibody in ANAs is more valuable in distinguishing immunity from viral and drug-induced liver diseases.

目的 探讨实时荧光核酸恒温扩增检测技术(simultaneous amplification and testing,SAT)检测人巨细胞病毒(human cytomegalovirus,HCMV)基质表层蛋白pp67(phosphoprotein67,pp67)mRNA在人类免疫缺陷病毒(human immunodeficiency virus,HIV)合并HCMV感染患者临床诊疗中的价值.方法 利用SAT技术构建检测HCMV pp67 mRNA的方法,评估SAT法检测HCMV pp67 mRNA重复性、特异性以及检测下限;收集HIV阳性HCMV活动性感染患者69例、HIV阳性HCMV非活动性/潜伏感染154例、HIV阴性HCMV非活动性/潜伏感染79例患者的血清、肺泡灌洗液、脑脊液等样本进行HCMV pp67 mRNA检测,评估特异度、灵敏度、正确率、阴性预测值、阳性预测值以及约登指数,评估HCMV pp67 mRNA在辨别HCMV活动性感染中的临床价值.结果 SAT法检测HCMV pp67 mRNA的精密度(变异系数<10％)可满足临床检测要求,与其他病原体无交叉反应,特异性良好,试剂检测下限为400 copies/mL;特异度87.55％,灵敏度60.87％,正确率81.46％,阴性预测值0.88,阳性预测值0.59以及约登指数0.48;SAT法检测HCMV pp67 mRNA结果阳性率与临床诊断结果差异无统计学意义(P=0.894).结论 SAT法检测HCMV pp67 mRNA可用于HCMV活动性感染的辅助诊断.

Copper nanoclusters (Cu NCs) as emerging luminescent metal NCs are gaining increasing attention owing to the comparatively low cost and high abundance of the Cu element in nature. However, it remains challenging to manipulate the optical properties of Cu NCs. Unlike most dispersed Cu NCs, whose luminescence efficiency was restricted by nonexcited relaxation, the Cu NCs confined in a porous poly-l-cysteine (poly-l-Cys) film were generated controllably with enhanced electrochemiluminescence (ECL) by in situ electrochemical reduction. Specifically, poly-l-Cys provided a porous structure to regulate the generation of Cu NCs within its holes, which not only increased the restriction on the intramolecular vibration and rotation of the ligands but also expedited the electron transfer near the electrode surface, reflecting in an enhancement of the ECL signal and efficiency. As an application of the confined Cu NCs, an ECL biosensor with high performance was constructed skillfully for highly sensitive detection of alkaline phosphatase (ALP), which adopted Cu NCs as the ECL luminophore and poly-l-Cys as a coreaction accelerator in a novel ECL ternary system (Cu NCs/S2O82-/poly-l-Cys). Furthermore, an ingenious target amplification based on the combination of a DNA walker and click chemistry was developed to convert ALP to DNA strands efficiently, achieving great improvement in the recognition efficiency. As a result, the biosensor had a low detection limit (9.5 × 10-7 U·L-1) and a wide linear range (10-8-10-2 U·L-1) for ALP detection, which showed great promise for the detection of non-nucleic acid targets and the diagnosis of diseases.

目的 评价全自动核酸检测平台在高敏HCV RNA定量检测的性能及检测临床样本的能力.方法 采用WHO国际标准品、HCV RNA假病毒样本、基因型血浆盘及临床样本,评价Pre-NATⅡ全自动核酸检测体系的正确度、精密度、线性范围、最低检出限/最低定量限、抗干扰能力等指标.结果 Pre-NATⅡ全自动核酸检测体系的精密度<5％,最低检出限为15 IU/mL,线性范围为30～1.0×109 IU/mL.检测WHO国际标准品的实测对数值和理论对数值偏差小于±0.20(-0.04～0.19),对临床常见的内源性干扰物、抗病毒药物和自身免疫疾病样本具有良好的抗干扰能力,且检测临床样本结果与临床常用试剂检测结果一致性(符合率为100％)和相关性(r>0.98)强.结论 Pre-NATⅡ全自动核酸检测体系准确性高、精密性好、定量范围宽、重复性好、抗干扰能力强,能够满足临床应用需求.

目前,国内新冠疫情防控取得阶段性胜利,但全球疫情形势愈发严峻,已进入全球大流行阶段.临床检验实验室检查在新型冠状病毒肺炎患者的诊疗中发挥重要的作用,对于有效控制疫情至关重要.本文对临床检验实验室在传染病疫情防控中发挥的作用进行讨论,提出建立平战结合的检验体系,提高应对突发传染性疾病的防控能力.

目的 验证人巨细胞病毒核酸定量检测试剂的分析性能是否能达到临床检测的需求.方法 依照《医学实验室质量和能力认可准则及其在分子诊断领域的应用说明》(CNAS-CL02-A009)文件要求,使用人巨细胞病毒核酸定量检测试剂盒质控品、标准品和卫生部临检中心室间质量评价质控品,对其可比性、精密度、检出下限以及线性进行性能验证和评估.结果 精密度验证,低浓度标本和高浓度标本的CV值(8.02％,1.19％)<10％,精密度符合临床要求.检出下限验证,重复检测浓度为500 copies/mL的样品10次,阳性检出率为100％,符合临床要求.可比性验证,通过率达100％,可比性符合要求.线性评估,R2=0.9954>0.95,验证合格.结论 通过验证,巨细胞病毒核酸定量检测可以满足巨细胞病毒筛查以及临床治疗效果的监测.

Background: Glioma is the most frequent primary malignant brain tumor, characterized by high morbidity, high mortality and dismal prognosis. Numerous analyses have revealed the abnormal expression of long non-coding RNAs (lncRNAs) in glioma cells. This study aims to explore its role in glioma development and prognosis. Methods: The gene expression in cell lines were measured by qRT-PCR. The role of LINC00663 in glioma was confirmed by CCK8, EdU assay, transwell and western blot as well as by in vivo experiments. Besides, Pearson's correlation analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were also performed as needed. Results: Firstly, data from our preliminary work (NSFC NO. 81572474) showed that LINC00663 might be largely implicated in glioma. Meanwhile, LINC00663 upregulation confirmed in glioma predicted poor clinical outcomes. Functionally, LINC00663 knockdown restrained cell proliferation, migration and invasion in vitro. Mechanistic investigations validated that LINC00663 silencing decreased AKT activation in glioma cells. Conclusions: LINC00663 promotes glioma development and progression through regulating AKT pathway, suggesting LINC00663 as a probable target for glioma treatment.

Aim: To investigate the anticancer effects of Jinlong capsule (JLC) against human glioblastoma cells and the possible underlying mechanism. Methods: Cell Counting Kit-8 and colony formation assay were adopted for the analysis of cell viability. Cell invasion and migration were evaluated by transwell and wound healing assays. Then, the expression level of mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), S6 and phosphorylated S6 (p-S6) were determined by western blotting. Results: The results showed that JLC significantly inhibited human glioblastoma cell proliferation, invasion and migration in a dose-dependent manner. The expressions of p-mTOR and p-S6 were dramatically suppressed by JLC. Furtherly, inhibition of mTOR reduced the cell migration and invasion, while the mTOR agonist (MHY1485) could partially reverse the anti-migration and anti-invasion activity of JLC. Conclusion: The above results suggested that JLC would be a potential candidate for the treatment of glioblastoma.