中药黑骨藤HPLC-ELSD指纹图谱研究

目的 建立中药黑骨藤HPLC-ELSD指纹图谱并结合聚类分析和主成分分析进行药材质量评价。方法 采用通用型ELSD检测器建立17个不同产地的黑骨藤药材的指纹图谱,采用HPLC-Q-TOF-MS技术对黑骨藤中化学成分进行表征,并通过指纹图谱相似度评价系统、层次聚类分析和主成分分析评价药材质量。结果 指纹图谱共标定了14个共有峰,采用液质联用技术共鉴定了58个化合物。除2批广西产地黑骨藤指纹图谱与对照图谱的相似度为0.663和0.826,其他15批产地的指纹图谱与对照图谱相似度均在0.9以上。指纹图谱相似度评价、层次聚类分析和主成分分析结果互相印证。结论 黑骨藤HPLC-ELSD指纹图谱研究结合聚类分析和主成分分析为该药材的真伪鉴别、质量控制提供了参考。

AcWRKY28 mediated activation of AcCPK genes confers salt tolerance in pineapple(Ananas comosus)

Unfavorable environmental cues severely affect crop productivity resulting in significant economic losses to farmers. In plants, multiple regulatory genes, such as the WRKY transcription factor (TF) family, modulate the expression of defense genes. However, the role of the pineapple WRKY genes is poorly understood. Here, we studied the pineapple WRKY gene, AcWRKY28, by generating AcWRKY28 over-expressing transgenic pineapple plants. Overexpression of AcWRKY28 enhanced the salt stress resistance in transgenic pineapple lines. Comparative transcriptome analysis of transgenic and wild-type pineapple plants showed that “plant-pathogen interaction” pathway genes, including 9calcium-dependent protein kinases (CPKs), were up-regulated in AcWRKY28 over-expressing plants. Furthermore, chromatin immunoprecipitation and yeast one-hybrid assays revealed AcCPK12, AcCPK3, AcCPK8, AcCPK1, and AcCPK15 as direct targets of AcWRKY28. Consistently, the study of AcCPK12 over-expressing Arabidopsis lines showed that AcCPK12 enhances salt, drought, and disease resistance. This study shows that AcWRKY28 plays a crucial role in promoting salt stress resistance by activating the expression of AcCPK genes.

Crossroads:Pathogenic role and therapeutic targets of neutrophil extracellular traps in rheumatoid arthritis

Rheumatoid arthritis(RA)is a prevalent autoimmune disease whose main features include chronic synovial inflammation,bone destruction,and joint degeneration.Neutrophils are often considered to be the first responders to inflammation and are a key presence in the inflammatory milieu of RA.Neutrophil extracellular traps(NETs),a meshwork of DNA-histone complexes and proteins released by activated neutrophils,are widely involved in the pathophysiology of autoimmune diseases,especially RA,in addition to playing a key role in the neutrophil innate immune response.NETs have been found to be an important source of citrullinated autoantigen antibodies and inflammatory factor release,which can activate RA synovial fibroblasts(FLS)and cause joint damage.This article reviews the role of NETs in the pathophysiology of RA,demonstrating the application of multiple molecules with various therapies,with a view to informing the discovery and development of novel biomarkers and therapeutic targets for RA.

基于HPLC指纹图谱和化学计量学的复方五凤草液的质量评价

目的建立复方五凤草液HPLC指纹图谱,结合化学计量学,测定成分含量,对不同产地泽漆的复方五凤草液进行质量评价。方法采用HiQ sil C_(18)W色谱柱(4.6 mm×250 mm,5μm),以甲醇-0.05%磷酸溶液为流动相,梯度洗脱;流速为1.0 mL·min^(-1);检测波长为270 nm;柱温为40℃;建立不同产地泽漆的复方五凤草液指纹图谱,进行相似度评价以及化学计量学分析,借助变量重要性投影值等方式筛选指认目标成分,进行含量测定。结果15批复方五凤草液指纹图谱含7个共有峰,所有样品与对照图谱相似度在0.795~0.999之间;经化学计量学分析,15批复方五凤草液聚为两类,复方五凤草液中没食子酸成分含量0.90~4.32 mg·g^(-1)。结论建立的指纹图谱分析和含量测定方法可靠、重复性好,可为复方五凤草液的质量控制提供参考。

Effects of π-conjugation-substitution on ESIPT process for oxazoline-substituted hydroxyfluorenes

Excited-state intramolecular proton transfer(ESIPT) molecules are broadly applied to UV absorbers, fluorescence sensing, and lighting materials. In previous work, the fluorescence colors of oxazoline-substituted hydroxyfluorenes and hydroxylated benzoxazole were diversified by adding the π-conjugation. There is intriguing that the mechanism of diversified fluorescence colors induced by ESIPT. Here, the density functional theory(DFT) and time-dependent DFT(TDDFT)are advised to identify the effects of π-conjugation on ESIPT and photophysical properties. The stabilized geometrical configurations, frontier molecular orbitals(FMOs) isosurfaces, and O–H stretching vibration frequency analysis demonstrate that PT processes are more active in S1state. Constructing the minimum energy pathways of ESIPT processes, we find that the calculated peak of enol and keto fluorescence of naphthoxazole(NO–OH) is distinctly bathochromic-shift relative to the oxazoline-substituted hydroxyfluorenes(Oxa–OH) configuration when adding π-conjugation-substitution, and it means that π-conjugation-substitution can diversify the fluorescence color. We hope our studies can establish new channels to devise the ESIPT-based molecules.

DNA methylation as a mediator of epigenetic regulation in the pathogenesis and precision medicine of osteoarthritis:An updated review

The pathophysiology of osteoarthritis(OA)is multifactorial,with the primary risk factors being obesity,age,environmental variables,and genetic predisposition.The available evidence suggests that genetic diversity does not adequately account for all clinical characteristics and heterogeneity of OA.Genetics has emerged as a nascent and crucial area of research in OA.The epigenetic module presents a potential link between genetic and environmental risk factors and the susceptibility and pathogenesis of OA.As a critical epigenetic alteration,DNA methylation has been shown to have an important role in the etiology of OA and is a viable biomarker for predicting disease progression and medication response,as shown in this research.This review aims to update knowledge in the field of DNA methylation associated with OA to better identify the essential features of OA subtypes and pathological conditions,hence accelerating individualized treatment and precision medicine.

Epigenetic modification mechanisms of chloroplasts mutants in pineapple leaves during somatic regeneration

Somaclonal variation in tissue culture is a common phenomenon induced by various external or internal environmental conditions,resulting in heritable or non-heritable alterations in gene expression. One crucial mechanism involved in plant growth and development is epigenetic regulation. A highly dynamic epigenome can respond to environmental changes by regulating gene expression. DNA methylation is one of these epigenetic modifications that can alter gene expression in tissue-cultured pineapple plants. The underlying mechanism of such somaclonal variations in pineapple and the epigenetic regulation involvement in somaclonal variations has not been studied. This study performed DNA methylome and transcriptome sequencing of wild-type(WT) and mutant pineapple plants(WS, HW, and TW). We observed altered DNA methylation patterns in chlorophyll development in the mutants. Specifically, we noticed that the methylation levels in the CHG and CHH contexts were lower in the gene body regions compared to the upstream and downstream regions. We identified several thousand differentially methylated regions(DMRs) located at the gene body regions, some of which overlapped with the differentially expressed genes(DEGs). Functional enrichment analyses suggested that these genes were involved in chlorophyll metabolism. Thus, our results revealed that the transcriptional regulation of many chlorophyll metabolic essential genes could be regulated by DNA methylation caused by somaclonal variations and provided insights into epigenetic mechanisms underlying the regulation of chlorosis in pineapple plants.

Exploration of the oxidative-inflammatory potential targets of Coicis Semen in osteoarthritis:Data mining and systematic pharmacology

On the basis of data mining,systematic pharmacology,molecular docking,and experiment validation,the oxidative-inflammatory molecular targets of Coicis Semen in the therapy of osteoarthritis(OA)were explored.Methods:The association rule analysis was effectively applied to highlight the correlation between Coicis Semen and oxidative inflammation indices.The random walk model was subsequently used to evaluate the clinical efficacy of Coicis Semen.Network pharmacology was used to predict network targets.The binding affinity of the active ingredient in Coicis Semen to the key target of OA was also successfully predicted.Results:Coicis Semen showed a significant reduction in oxidative-inflammatory indicators of OA.A total of 108 promising targets were predicted for the 24 bioactive compounds in Coicis Semen.Eight target genes were considered core target genes.The enrichment analysis predicts that Coicis Semen may activate the interleukin(IL)-17,mitogen-activated protein kinase(MAPK),and nuclear factor kappa B(NF-kappa B)signaling pathways.Molecular docking demonstrated that stigmasterol,2-monoolein,sitosterol,and sitosterol alpha1 had free binding energies to oxidative and inflammatory targets(MAPK1,Estrogen Receptor 1[ESR1],and Peroxisome Proliferator-Activated Receptor Alpha[PPARA]).Both clinical trials and in vitro cell experiments revealed that Coicis Semen could increase ESR1 and PPAR-αlevels while decreasing MAPK1 levels.Conclusions:Coicis Semen has a remarkable anti-OA effect.Precisely,the major components of Coicis Semen,including stigmasterol,sitosterol alpha1,sitosterol,and 2-monoolein,specifically inhibit MAPK1,ESR1,and PPARA to reduce the inflammatory response and oxidative damage in OA.

浅析叉车护顶架的定期检验以及设置

本文介绍了护顶架在叉车使用中的重要性,讲述了护顶架在检规中逐渐被重视的过程,着重介绍了《场(厂)内专用机动车辆安全技术规程》(TSG 81—2022)(以下简称新检规)在执行中的疑惑点;通过对国标的分析,阐述了叉车护顶架的设置要求,以及检规中护顶架的相关条款与国标的对应情况,提出了相应的检验方法和一些建议。

Cr基过渡层对CrWN涂层摩擦学性能的影响

本文探究了Cr过渡层及Cr-CrN过渡层对CrWN涂层耐磨性能的影响,制备具有良好的减磨耐磨性能的CrWN涂层。采用等离子增强磁控溅射工艺,在碳化钨基体表面制备CrWN涂层,Cr-CrWN涂层、Cr-CrN-CrWN涂层。通过扫描电子显微镜,X射线光电子能谱仪,X射线衍射仪,纳米压痕仪,洛氏硬度计和摩擦磨损实验机对涂层表面形貌、力学性能及摩擦磨损性能进行表征及分析。三种CrWN涂层均以面心立方结构的CrN相和W2N相构成。Cr-CrN-CrWN涂层的膜基结合强度最高,为HF1等级。CrWN涂层的硬度为21.5 GPa,引入Cr过渡层及Cr-CrN过渡层均未对涂层硬度及弹性模量造成显著影响。但通过摩擦磨损实验结果表明Cr过渡层及Cr-CrN过渡层均可降低涂层的摩擦系数并提高涂层的耐磨损性能,在稳定摩擦阶段CrCrWN涂层和Cr-CrN-CrWN涂层的摩擦系数分别为0.656,0.703,磨损率分别为1.366×10^(-6)mm^(3)/(N·m)、1.247×10^(-6)mm^(3)/(N·m);且磨损后磨痕表面无犁沟及裂纹,涂层未出现明显剥落。因此,Cr过渡层及Cr-CrN梯度过渡层可改善CrWN涂层的摩擦学性能及结合强度,并提高涂层的耐磨损性能。

The evolving Gleason grading system

The Gleason grading system for prostate adenocarcinoma has evolved from its original scheme established in the 1960s–1970s, to a significantly modified system after two major consensus meetings conducted by the International Society of Urologic Pathology(ISUP) in 2005 and 2014, respectively. The Gleason grading system has been incorporated into the WHO classification of prostate cancer, the AJCC/UICC staging system, and the NCCN guidelines as one of the key factors in treatment decision. Both pathologists and clinicians need to fully understand the principles and practice of this grading system. We here briefly review the historical aspects of the original scheme and the recent developments of Gleason grading system, focusing on major changes over the years that resulted in the modern Gleason grading system, which has led to a new "Grade Group" system proposed by the 2014 ISUP consensus, and adopted by the 2016 WHO classification of tumours of the prostate.

Intraductal carcinoma of prostate(IDC-P): from obscure to significant

The concept of intraductal carcinoma of prostate(IDC-P) has evolved over the years and its clinicopathologic significance has come to be more clearly appreciated. In contrast to morphologically malignant intraductal lesions that represent earlier stages of the malignant process in other anatomic sites such as the breast, IDC-P has now been generally recognized as a prognostically unfavorable manifestation of later stage spreading of its invasive counterpart. We here briefly review the evolution of the IDC-P concept, the histological diagnostic criteria and differential diagnosis, the clinical significance, as well as recent molecular data of IDC-P.

Effects of chemotherapeutic agent bendamustine for nonhodgkin lymphoma on spermatogenesis in mice

Non-Hodgkin lymphoma(NHL) is one of the most common cancers affecting men of reproductive age. The high response rate of bendamustine as first-line treatment for NHL, coupled with young age of patients, makes elucidation of the impact of treatment on male reproduction important. Our aim was to determine the effects of bendamustine on male reproduction by animal model. Male mice were treated with bendamustine(40 mg/kg) through tail vein injection while cisplatin was given as a standard(3 mg/kg) through intraperitoneal injection. After 3 weeks, bendamustine induced weight loss and sperm morphology abnormalities were compared to the control. Additionally, sperm with folded tails were the most frequent abnormality in bendamustine-treated mice. But the mechanism of sperm abnormality induced by bendamustine remains to be elucidated. These results indicate bendamustine may affect spermatozoa of patients who have been treated for NHL.

Theoretical investigation of fluorescence changes caused by methanol bridge based on ESIPT reaction

The different fluorescence behavior caused by the excited state proton transfer in 3-hydroxy-4-pyridylisoquinoline(2a)compound has been theoretically investigated.Our calculation results illustrate that the 2a monomer in tetrahydrofuran solvent would not occur proton transfer spontaneously,while the 2a complex in methanol(MeOH)solvent can undergo an asynchronous excited state intramolecular proton transfer(ESIPT)process.The result was confirmed by analyzing the related structural parameters,infrared vibration spectrum and reduced density gradient isosurfaces.Moreover,the potential curves revealed that with the bridging of single MeOH molecular the energy barrier of ESIPT was modulated effectively.It was distinctly reduced to 4.80 kcal/mol in 2a-MeOH complex from 25.01 kcal/mol in 2a monomer.Accordingly,the ESIPT process induced a fluorochromic phenomenon with the assistant of proton-bridge.The elucidation of the mechanism of solvent discoloration will contribute to the design and synthesis of fluorogenic dyes as environment-sensitive probes.

Tumor-to-tumor metastasis of clear cell renal cell carcinoma to contralateral synchronous pheochromocytoma:A case report

BACKGROUND Tumor-to-tumor metastasis(TTM)is an uncommon condition.Only a few cases of renal cell carcinoma(RCC)as donor tumor of TTM have been reported in literature,and none of these studies have described RCC metastasizing to synchronous pheochromocytoma(PCC).CASE SUMMARY The patient was a 54-year-old woman who presented with recurrent dull abdominal pain for six months,which was further aggravated for one more month.Enhanced computed tomography revealed a tumor mass in the right kidney and another mass in the left retroperitoneum/adrenal gland.Histopathology and immunochemistry of resected specimens confirmed the diagnosis of clear cell renal cell carcinoma(CCRCC)of the right kidney,and the left retroperitoneum revealed a typical PCC with CCRCC metastasis.Whole exome sequencing revealed the presence of a c.529A>T somatic mutation of the Von Hippel Lindau(VHL)gene in the metastasized CCRCC,which was also present in the primary right kidney CCRCC,as confirmed by Sanger sequencing.No VHL mutation was detected in the PCC or in normal right kidney tissue.Fluorescence in situ hybridization revealed loss of chromosome 3p in both the primary right kidney CCRCC and CCRCC metastasized to PCC in the left kidney.CONCLUSION This is the first case showing metastasis of CCRCC to PCC,thus leading to tumorto-tumor metastasis.

Theoretical study on the mechanism for the excited-state double proton transfer process of an asymmetric Schiff base ligand

Excited-state double proton transfer(ESDPT)in the 1-[(2-hydroxy-3-methoxy-benzylidene)-hydrazonomethyl]-naphthalen-2-ol(HYDRAVH_(2))ligand was studied by the density functional theory and time-dependent density functional theory method.The analysis of frontier molecular orbitals,infrared spectra,and non-covalent interactions have crossvalidated that the asymmetric structure has an influence on the proton transfer,which makes the proton transfer ability of the two hydrogen protons different.The potential energy surfaces in both S_(0)and S_1 states were scanned with varying O-H bond lengths.The results of potential energy surface analysis adequately proved that the HYDRAVH_(2)can undergo the ESDPT process in the S_1 state and the double proton transfer process is a stepwise proton transfer mechanism.Our work can pave the way towards the design and synthesis of new molecules.

Effects of electroacupuncture of acupoints of the neiguan and ximen on ischemic myocardium energy metabolism with acute myocardial ischemia rats

Objective:To observe the effect of Electroacupuncture of Acupoints of the Neiguan(PC6)and Ximen(PC4)on its myocardial energy metabolism with acute myocardial ischemia rats caused by coronary artery ligation.Methods:Forty standard Spargue-Dawely(SD)rats were divided into four groups randomly,namely,group of Neiguan,group of Ximen,model group,sham group.The group of rats of myocardial ischemia were induced by coronary ligation,and the model rat of acute myocardial ischemia was prepared.The sham group had no ligation of the needle.After anesthesia was awakened,acupuncture was applied bilateral at Neiguan and Ximen of the group of Neiguan and Ximen,once a day at a fixed time for 14 days.The rats of model group and the sham group were not given acupunctured,but use the samely methods of Neiguan and Ximen groups to grap the rats at the same time everyday.After 14 days the experiment was over,observed the pathological changes of myocardial tissue by HE staining,detected the levels of free fatty acid(FFA),adenosine triphosphate(ATP)content,and the activities of Na^(+)-K^(+)-ATPase according to the reagents´demand.Results:Pathological results showed that acupuncuted at Neiguan and Ximen could improve the ischemic cardiomyocyte injury.Compared with the sham group,the levels of FFA in model group were increased(p<0.01),the levels of ATP,the activities of Na^(+)-K^(+)-ATPase were all decreased(p<0.01);Compared with the model group,the levels of FFA were decreased(p<0.05,p<0.01),the levels of ATP,the activities of Na^(+)-K^(+)-ATPase were improved.Conclusion:Acupuncturing at the Pericardium Meridian can alleviate the cell injury by myocardial ischemia,and also can improve the myocardial energetic metabolism in acute myocardial ischemia.

Performance Comparison of Computational Methods for the Prediction of the Function and Pathogenicity of Non-coding Variants

Non-coding variants in the human genome significantly influence human traits and complex diseases via their regulation and modification effects.Hence,an increasing number of computational methods are developed to predict the effects of variants in human non-coding sequences.However,it is difficult for inexperienced users to select appropriate computational methods from dozens of available methods.To solve this issue,we assessed 12 performance metrics of 24 methods on four independent non-coding variant benchmark datasets:(1)rare germline variants from clinical relevant sequence variants(ClinVar),(2)rare somatic variants from Catalogue Of Somatic Mutations In Cancer(COSMIC),(3)common regulatory variants from curated expression quantitative trait locus(eQTL)data,and(4)disease-associated common variants from curated genomewide association studies(GWAS).All 24 tested methods performed differently under various conditions,indicating varying strengths and weaknesses under different scenarios.Importantly,the performance of existing methods was acceptable for rare germline variants from ClinVar with the area under the receiver operating characteristic curve(AUROC)of 0.4481–0.8033 and poor for rare somatic variants from COSMIC(AUROC=0.4984–0.7131),common regulatory variants from curated eQTL data(AUROC=0.4837–0.6472),and disease-associated common variants from curated GWAS(AUROC=0.4766–0.5188).We also compared the prediction performance of 24 methods for non-coding de novo mutations in autism spectrum disorder,and found that the combined annotation-dependent depletion(CADD)and context-dependent tolerance score(CDTS)methods showed better performance.Summarily,we assessed the performance of 24 computational methods under diverse scenarios,providing preliminary advice for proper tool selection and guiding the development of new techniques in interpreting non-coding variants.

Identification of methylated differentially expressed hub genes in systemic lupus erythematosus patients by WGCNA-based epigenome-wide and transcriptome-wide analysis

Systemic lupus erythematosus(SLE)is an autoimmune disease,characterized by the production of autoantibodies and the involvement of multi-systems.In order to explore its molecular mechanism,we,using bioinformatics analysis and wet lab experiments,identified two key genes in SLE patients.

参松养心胶囊多组分同时测定及方法学验证

目的:建立高效液相色谱法同时测定参松养心胶囊中8个主要成分莫诺苷、马钱苷、芍药苷、丹酚酸B、五味子酯甲、丹参酮Ⅰ、五味子甲素和丹参酮Ⅱ_A的含量并对方法学进行验证。方法:以L_9(3~4)正交试验优化提取条件,采用50%甲醇超声提取75 min,料液比为0.4 g∶10 mL。液相色谱分析采用ODS-2 C_(18)色谱柱(4.6 mm×250 mm,5μm),柱温30℃,以甲醇-乙腈(1∶1)及甲酸溶液(pH3)为流动相,梯度洗脱,流速1.1 mL·min^(-1),检测波长230 nm。结果:莫诺苷、马钱苷、芍药苷、丹酚酸B、五味子酯甲、丹参酮Ⅰ、五味子甲素和丹参酮Ⅱ_A分别在19.84～198.40μg·mL^(-1)(r=0.999 0)、10.20～101.98μg·mL^(-1)(r=0.999 3)、26.76～267.62μg·mL^(-1)(r=0.999 4)、94.32～943.24μg·mL^(-1)(r=0.999 6)、4.50～45.01μg·mL^(-1)(r=0.999 5)、4.95～49.45μg·mL^(-1)(r=0.999 9)、3.11～31.12μg·mL^(-1)(r=0.999 5)和2.96～29.61μg·mL^(-1)(r=0.999 5)范围内呈良好线性关系,平均加样回收率在100.3%～109.1%之间。10批样品中上述8个成分的含量范围分别为1.548～2.113、1.455～1.641、2.523～3.706、5.535～13.255、0.654～0.866、0.168～0.429、0.329～0.496和0.216～0.407 mg·g^(-1)。结论:本研究建立的方法可以用于中成药参松养心胶囊的多组分同时测定。