Role of TNF-related apoptosis-inducing ligand （TRAIL） in the pathogenesis of varicocele-induced testicular dysfunction

The higher frequency of varicocele in men with infertility has drawn attention and resulted in increased research at the molecular level towards treatments. The aim of this study was to investigate the role of tumor necrosis factor （TNF）-related apoptosis-inducing ligand （TRAIL） and its receptors in varicocele-induced testicular dysfunction in an experimental rat model. The rats were divided into three groups： control, sham and varicocele. Varicoceles in rats were induced by partial ligation of the left renal vein and left testes. The rats were analyzed 13 weeks after surgery. The degree of DNA fragmentation within cells in the testis was determined using terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling （TUNEL） assay. Tubule degeneration was evaluated using the Johnsen score. The expression of TRAIL and its receptors was detected by immunohistochemical and Western blotting techniques. The apoptotic index, Johnsen score and the expression of TRAIL and TRAIL receptors were examined. The data are presented as the mean-.＋s.d, and were analyzed using computer software. The KruskaI-Wallis and Dunn＇s multiple comparison tests were used in the statistical analyses. The germ cell apoptotic index was increased in rats with varicoceles when compared with the sham and control groups （P=0.0031）. The Johnsen score was significantly decreased in the varicocele group when compared with the sham and control groups （P〈O.O001）. Immunohistochemical and Western blotting analyses showed that after varicocele induction, the expression of TRAIL-R1 and TRAIL-R4 in germ cells was increased and the expression of TRAIL-R2 was decreased. There are no significant differences among the groups in terms of TRAIL and TRAIL-R3 receptor expression. The results of this study indicate that TRAIL and its receptors may have a potential role in the pathogenesis of varicocele-induced testicular dysfunction.

Comprehensive analysis of the potential pathogenesis of COVID-19 infection and liver cancer

BACKGROUND A growing number of clinical examples suggest that coronavirus disease 2019(COVID-19)appears to have an impact on the treatment of patients with liver cancer compared to the normal population,and the prevalence of COVID-19 is significantly higher in patients with liver cancer.However,this mechanism of action has not been clarified.Gene sets for COVID-19(GSE180226)and liver cancer(GSE87630)were obtained from the Gene Expression Omnibus database.After identifying the common differentially expressed genes(DEGs)of COVID-19 and liver cancer,functional enrichment analysis,protein-protein interaction network construction and scree-ning and analysis of hub genes were performed.Subsequently,the validation of the differential expression of hub genes in the disease was performed and the regulatory network of transcription factors and hub genes was constructed.RESULTS Of 518 common DEGs were obtained by screening for functional analysis.Fifteen hub genes including aurora kinase B,cyclin B2,cell division cycle 20,cell division cycle associated 8,nucleolar and spindle associated protein 1,etc.,were further identified from DEGs using the“cytoHubba”plugin.Functional enrichment analysis of hub genes showed that these hub genes are associated with P53 signalling pathway regulation,cell cycle and other functions,and they may serve as potential molecular markers for COVID-19 and liver cancer.Finally,we selected 10 of the hub genes for in vitro expression validation in liver cancer cells.CONCLUSION Our study reveals a common pathogenesis of liver cancer and COVID-19.These common pathways and key genes may provide new ideas for further mechanistic studies.

COPD患者血清TRAIL表达水平及与炎症因子和肺功能的相关性

目的探讨慢性阻塞性肺疾病(COPD)患者血清肿瘤坏死因子相关凋亡诱导配体(TRAIL)与炎症因子和肺功能的关系。方法选取2020年6月—2022年12月在本院治疗的200例COPD患者为观察组,200例健康体检志愿者为对照组。根据COPD诊治指南中疾病类型分为急性加重期组140例,稳定期组60例。分析血清TRAIL水平与炎症因子[超敏-C反应蛋白(hs-CRP)、降钙素原(PCT)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)]水平、肺功能[1秒用力呼气容积占比(FEV_(1)%)、FEV_(1)和用力肺活量(FVC)比值、肺一氧化碳弥散量占比(DLCO%)及呼气峰流速(PEF)]的关系。结果与对照组相比,稳定期组、急性加重期组患者血清TRAIL、hs-CRP、PCT、IL-6、TNF-α水平显著升高(P<0.05),FEV_(1)%、FEV_(1)/FVC、DLCO%、PEF明显降低(P<0.05);急性加重期组较稳定期组患者血清TRAIL、hs-CRP、PCT、IL-6、TNF-α水平明显升高(P<0.05),FEV_(1)%、FEV_(1)/FVC、DLCO%、PEF显著降低(P<0.05);多因素线性回归分析结果显示,TRAIL水平与hs-CRP、TNF-α、FEV_(1)/FVC和DLCO水平有关(P<0.05);经Pearson法分析,COPD患者血清TRAIL水平与hs-CRP、TNF-α均呈正相关(r=0.412,0.527,P<0.05),与FEV_(1)/FVC、DLCO%均呈负相关(r=-0.594,-0.476,P<0.05)。结论COPD患者血清TRAIL水平升高,其与患者炎症因子和肺功能密切相关。

Identification of hub genes associated with Helicobacter pylori infection and type 2 diabetes mellitus:A pilot bioinformatics study

BACKGROUND Helicobacter pylori(H.pylori)infection is related to various extragastric diseases including type 2 diabetes mellitus(T2DM).However,the possible mechanisms connecting H.pylori infection and T2DM remain unknown.AIM To explore potential molecular connections between H.pylori infection and T2DM.METHODS We extracted gene expression arrays from three online datasets(GSE60427,GSE27411 and GSE115601).Differentially expressed genes(DEGs)commonly present in patients with H.pylori infection and T2DM were identified.Hub genes were validated using human gastric biopsy samples.Correlations between hub genes and immune cell infiltration,miRNAs,and transcription factors(TFs)were further analyzed.RESULTS A total of 67 DEGs were commonly presented in patients with H.pylori infection and T2DM.Five significantly upregulated hub genes,including TLR4,ITGAM,C5AR1,FCER1G,and FCGR2A,were finally identified,all of which are closely related to immune cell infiltration.The gene-miRNA analysis detected 13 miRNAs with at least two gene cross-links.TF-gene interaction networks showed that TLR4 was coregulated by 26 TFs,the largest number of TFs among the 5 hub genes.CONCLUSION We identified five hub genes that may have molecular connections between H.pylori infection and T2DM.This study provides new insights into the pathogenesis of H.pylori-induced onset of T2DM.

Factors determining changes in the network of marked hiking trails in the Sudetes

Hiking trails are a basic type of tourist infrastructure,which,on the one hand,make areas available for tourist traffic,and on the other hand,can contribute to the protection of the natural environment(if they are well designed and maintained).Owing to the variety of performed functions,their designation is determined by several factors:natural,technical,economic,social.Networks of trails change constantly.The aim of this article is to determine exactly what factors influence transformations within the hiking trail networks and what is their significance.To this end,three study areas in the Sudetes were analysed:one on the Polish side of the Sudetes-the Table(Stołowe)Mountains,and two on the Czech side-the rock town near the village of Sloup vČechách and the central part of Zlatohorskávrchovina.An analysis of changes in the shape of the networks over time was carried out,as well as surveys of institutions that were responsible for or influenced these transformations.These areas are characterised by a significant level of changes in the trail network.Among the factors influencing these changes,the tourist attractiveness of the area,the resilience of the environment,the intensity of tourism traffic,the environmental transformations associated with it,the history of tourism development and land ownership changes should be considered the most important.At each stage of forming networks,the key factor should be tourists’needs,including the desire to escape the urbanised environment.For this reason,trails should avoid roads with artificial(hard)surfaces and heavy automobile traffic.

Key genes and regulatory networks for diabetic retinopathy based on hypoxia-related genes:a bioinformatics analysis

AIM:To prevent neovascularization in diabetic retinopathy(DR)patients and partially control disease progression.METHODS:Hypoxia-related differentially expressed genes(DEGs)were identified from the GSE60436 and GSE102485 datasets,followed by gene ontology(GO)functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.Potential candidate drugs were screened using the CMap database.Subsequently,a protein-protein interaction(PPI)network was constructed to identify hypoxia-related hub genes.A nomogram was generated using the rms R package,and the correlation of hub genes was analyzed using the Hmisc R package.The clinical significance of hub genes was validated by comparing their expression levels between disease and normal groups and constructing receiver operating characteristic curve(ROC)curves.Finally,a hypoxia-related miRNA-transcription factor(TF)-Hub gene network was constructed using the NetworkAnalyst online tool.RESULTS:Totally 48 hypoxia-related DEGs and screened 10 potential candidate drugs with interaction relationships to upregulated hypoxia-related genes were identified,such as ruxolitinib,meprylcaine,and deferiprone.In addition,8 hub genes were also identified:glycogen phosphorylase muscle associated(PYGM),glyceraldehyde-3-phosphate dehydrogenase spermatogenic(GAPDHS),enolase 3(ENO3),aldolase fructose-bisphosphate C(ALDOC),phosphoglucomutase 2(PGM2),enolase 2(ENO2),phosphoglycerate mutase 2(PGAM2),and 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3(PFKFB3).Based on hub gene predictions,the miRNA-TF-Hub gene network revealed complex interactions between 163 miRNAs,77 TFs,and hub genes.The results of ROC showed that the except for GAPDHS,the area under curve(AUC)values of the other 7 hub genes were greater than 0.758,indicating their favorable diagnostic performance.CONCLUSION:PYGM,GAPDHS,ENO3,ALDOC,PGM2,ENO2,PGAM2,and PFKFB3 are hub genes in DR,and hypoxia-related hub genes exhibited favorable diagnostic performance.

Identification and Validation of Vascular-Associated Biomarkers for the Prognosis and Potential Pathogenesis of Hypertension Using Comprehensive Bioinformatics Methods

Background: Hypertension, also known as increased blood pressure, is a phenomenon in which blood flows in blood vessels and causes persistently higher-than-normal pressure on the vessel wall. The identification of novel prognostic and pathogenesis biomarkers plays a key role in the management of hypertension. Methods: The GSE7483 and GSE75815 datasets from the gene expression omnibus (GEO) database were used to identify the genes associated with hypertension that were differentially expressed genes (DEGs). The functional role of the DEGs was elucidated by gene body (GO) enrichment analysis. In addition, we performed an immune infiltration assay and GSEA on the DEGs of hypertensive patients and verified the expression of novel DEGs in the blood of hypertensive patients by RT-qPCR. Results: A total of 267 DEGs were identified from the GEO database. GO analysis revealed that these genes were associated mainly with biological processes such as fibroblast proliferation, cell structural organization, extracellular matrix organization, vasculature development regulation, and angiogenesis. We identified five possible biomarkers, Ecm1, Sparc, Sphk1, Thbsl, and Mecp2, which correlate with vascular development and angiogenesis characteristic of hypertension by bioinformatics, and explored the clinical expression levels of these genes by RT-qPCR, and found that Sparc, Sphk1, and Thbs1 showed significant up-regulation, in agreement with the results of the bioinformatics analysis. Conclusion: Our study suggested that Sparc, Sphk1 and Thbs1 may be potential novel biomarkers for the diagnosis, treatment and prognosis of hypertension and that they are involved in the regulation of vascular development and angiogenesis in hypertension.

Mining elite loci and candidate genes for root morphology-related traits at the seedling stage by genome-wide association studies in upland cotton(Gossypium hirsutum L.)

Root system architecture plays an essential role in water and nutrient acquisition in plants,and it is significantly involved in plant adaptations to various environmental stresses.In this study,a panel of 242 cotton accessions was collected to investigate six root morphological traits at the seedling stage,including main root length(MRL),root fresh weight(RFW),total root length(TRL),root surface area(RSA),root volume(RV),and root average diameter(AvgD).The correlation analysis of the six root morphological traits revealed strong positive correlations of TRL with RSA,as well as RV with RSA and AvgD,whereas a significant negative correlation was found between TRL and AvgD.Subsequently,a genome-wide association study(GWAS)was performed using the root phenotypic and genotypic data reported previously for the 242 accessions using 56,010 single nucleotide polymorphisms(SNPs)from the CottonSNP80K array.A total of 41 quantitative trait loci(QTLs)were identified,including nine for MRL,six for RFW,nine for TRL,12 for RSA,12 for RV and two for AvgD.Among them,eight QTLs were repeatedly detected in two or more traits.Integrating these results with a transcriptome analysis,we identified 17 candidate genes with high transcript values of transcripts per million(TPM)≥30 in the roots.Furthermore,we functionally verified the candidate gene GH_D05G2106,which encodes a WPP domain protein 2in root development.A virus-induced gene silencing(VIGS)assay showed that knocking down GH_D05G2106significantly inhibited root development in cotton,indicating its positive role in root system architecture formation.Collectively,these results provide a theoretical basis and candidate genes for future studies on cotton root developmental biology and root-related cotton breeding.

PbrARF4 contributes to calyx shedding of fruitlets in ‘Dangshan Suli’ pear by partly regulating the expression of abscission genes

Fruitlet calyx shedding in pear plants is apparently regulated via numerous pathways that involve both environmental triggers and phytohormones cues such as auxin. In this study, we found at 10 days after full bloom (DAFB) higher levels of indoleacetic acid (IAA) and tryptophan (Trp) in calyx persistence fruitlet (CPF) than calyx shedding fruitlet (CSF) ofDanshan Suli’ pear (Pyrus bretschneideri Rhed.). Consisting with this, the activity of indolealdehyde oxidase (IAAIdO), which promotes IAA synthesis, was remarkably increased, and that of peroxidase(POD), which degrades IAA, dropped markedly in CPF but not in CSF. Further, qRT-PCR results revealed that most of 31 PbrARFs (encoding auxin response factors) in Pyrus bretschneideri were highly expressed in CPF, whereas PbrARF4, PbrARF24 and PbrARF26 were significantly downregulated in CPF vis-a-vis CSF. Phylogenetic analysis revealed that 6 PbrARFs clustered in the group III, where PbrARF4 showed the closest affinity with AtARF1 that promotes organ abscission, indicating a putative role of PbrARF4 in mediating the process of calyx shedding in pear. In fact, the ectopic overexpression of PbrARF4 in Solanum lycopersicum resulted in an earlier-formed and deeper abscission layer (AL) in the transgenic plants, whose calyxes were more prone to wilt at the mature red stage (MR) compared with the control plants (wild-type). More importantly, expression levels of the abscission genes SILS and Sl Cel2 in transgenic plants overexpressing PbrARF4 were significantly upregulated in comparation with the WT, whereas those of Sl BI and Sl TAPG2 were considerably inhibited. Further, PbrJOINTLESS and PbrIDA,the two genes related to calyx shedding in pear, were up-regulated more in CSF than CPF. The findings contribute to a better understanding of PbrARFs involved in fruitlet calyx shedding of pear, which could prove beneficial to improving the quality of pear fruit.

子痫前期患者血浆中sTRAIL、胎盘组织TRAIL及TRAIL受体的变化及意义

目的:研究子痫前期(PE)患者血浆s TRAIL及胎盘滋养细胞上TRAIL/TRAIL-Rs的变化及其与PE发病的关系。方法:选取56例PE患者为研究对象,其中符合重度PE诊断标准者31例(研究组1),未及重度PE 25例(研究组2)。选取同期正常妊娠妇女为对照组,其中28~36周30例(对照组1)、36^(+1)~38周30例(对照组2),因胎膜早破、早产等于28~36周分娩者27例(对照组3),36^(+1)~38周分娩者30例(对照组4)。研究组孕妇留取静脉血及胎盘组织,对照组1、2留取静脉血,对照组3、4留取胎盘组织。ELISA法检测血浆中TRAIL含量,免疫组化法检测胎盘组织中TRAIL和TRAIL-Rs(DR4、DR5、DcR1、DcR2)含量。结果:研究组血浆中sTRAIL含量明显低于对照组(研究组1 vs对照组1,P<0.001;研究组2 vs对照组2,P<0.005);随着病情加重,s TRAIL含量明显下降(P<0.05)。研究组胎盘组织中TRAIL表达明显减少,随着病情加重而加重。研究组中死亡受体DR4、DR5的含量明显高于对照组,而诱骗受体DcR1、DcR2含量明显低于对照组。PE重度组中DR4、DR5含量高于轻度组(P<0.05),DcR1和DcR2含量则无显著差异。结论:PE患者血浆中sTRAIL及胎盘组织中TRAIL/TRAIL-Rs含量发生了明显变化,TRAIL/TRAIL-Rs的不平衡性参与了PE的病理过程。

TRAIL受体在肿瘤细胞系上的表达及意义

目的 检测ＴＮＦ相关凋亡诱导配体（ＴＲＡＩＬ）的受体，在来源于血液系统、肝脏、肺脏和大肠的８个肿瘤细胞系中的表达，并探讨其意义。方法采用半定量ＲＴ－ＰＣＲ，对ＴＲＡＩＬ受体的表达进行半定量检测。结果 ＴＲＡＩＬ凋亡通路中，能够诱导凋亡反应的死亡受体ＤＲ４和ＤＲ５，在所检测的肿瘤细胞系中都有表达，其中ＤＲ５在所有肿瘤细胞系中的表达水平均显著高于ＤＲ４（Ｐ＜０．０５）。而能够竞争性阻断ＴＲＡＩＬ诱导的凋亡反应的诱骗受体ＤｃＲ１和ＤｃＲ２，在所有的肿瘤细胞中都呈低水平表达或不表达。结论 ＤＲ５可能在ＴＲＡＩＬ诱导凋亡的通路中发挥最重要的作用。ＴＲＡＩＬ死亡受体和诱骗受体在肿瘤细胞系中的表达具有差异性，这种差异性可在一定程度上解释不同细胞对ＴＲＡＩＬ诱导凋亡的敏感度。

全人源抗TRAIL-R1／-R2单克隆抗体肿瘤治疗的研究进展

目前全人源TRAIL受体（TRAIL-Rs）的单克隆抗体（单抗）对肿瘤靶向治疗已进入临床Ⅰ～Ⅳ期研究。TRAIL-Rs单抗可分别与死亡受体TRAIL-R1和TRAIL-R2结合，形成死亡诱导信号复合体（DISC），诱导肿瘤细胞凋亡，成为抗肿瘤抗体药物研发的热点之一。全人源TRAIL-Rs单抗与放疗、化疗药物或其它制剂以及免疫治疗方法联合应用在肿瘤治疗中取得了显著的效果，为临床抗肿瘤治疗提供了新方案。

Fate and Behavior of Tetracycline Resistance Genes in Activated Carbon Adsorption

The accessibility of tetracycline resistance gene (tetG) into the pores of activated carbon (AC), as well as the impact of the pore size distribution (PSD) of AC on the uptake capacity of tetG, were investigated using eight types of AC (four coal-based and four wood-based). AC showed the capability to admit tetG and the average reduction of tetG for coal-based and wood-based ACs at the AC dose of 1 g·L-1 was 3.12 log and 3.65 log, respectively. The uptake kinetic analysis showed that the uptake of the gene followed the pseudo-second-order kinetics reaction, and the uptake rate constant for the coal-based and wood-based ACs was in the range of 5.97 × 10-12 - 4.64 × 10-9 and 7.02 × 10-11 - 1.59 × 10-8 copies·mg-1·min-1, respectively. The uptake capacity analysis by fitting the obtained experiment data with the Freundlich isotherm model indicated that the uptake constant (KF) values were 1.71 × 103 - 8.00 × 109 (copies·g-1)1-1/n for coal-based ACs and 7.00 × 108 - 3.00 × 1010 (copies·g-1)1-1/n for wood-based ones. In addition, the correlation analysis between KF values and pore volume as well as pore surface at different pore size regions of ACs showed that relatively higher positive correlation was found for pores of 50 - 100 Å, suggesting ACs with more pores in this size region can uptake more tetG. The findings of this study are valuable as reference for optimizing the adsorption process regarding antibiotic resistance-related concerns in drinking water treatment.

The Application of Nicotiana benthamiana as a Transient Expression Host to Clone the Coding Sequences of Plant Genes

Coding sequences (CDS) are commonly used for transient gene expression, in yeast two-hybrid screening, to verify protein interactions and in prokaryotic gene expression studies. CDS are most commonly obtained using complementary DNA (cDNA) derived from messenger RNA (mRNA) extracted from plant tissues and generated by reverse transcription. However, some CDS are difficult to acquire through this process as they are expressed at extremely low levels or have specific spatial and/or temporal expression patterns in vivo. These challenges require the development of alternative CDS cloning technologies. In this study, we found that the genomic intron-containing gene coding sequences (gDNA) from Arabidopsis thaliana, Oryza sativa, Brassica napus, and Glycine max can be correctly transcribed and spliced into mRNA in Nicotiana benthamiana. In contrast, gDNAs from Triticum aestivum and Sorghum bicolor did not function correctly. In transient expression experiments, the target DNA sequence is driven by a constitutive promoter. Theoretically, a sufficient amount of mRNA can be extracted from the N. benthamiana leaves, making it conducive to the cloning of CDS target genes. Our data demonstrate that N. benthamiana can be used as an effective host for the cloning CDS of plant genes.

Remodeling tumor microenvironment using pH-sensitive biomimetic co-delivery of TRAIL/R848 liposomes against colorectal cancer

Background:Despite significant advancements in the development of anticancer therapies over the past few decades,the clinical management of colorectal cancer remains a challenging task.This study aims to investigate the inhibitory effects of cancer-targeting liposomes against colorectal cancer.Materials and Methods:Liposomes consisting of 3β-[N-(N′,N′-dimethylamino ethane)carbamoyl]-cholesterol(DC-CHOL),cholesterol(CHOL),and dioleoylphosphatidylethanolamine(DOPE)at a molar ratio of 1:1:0.5 were created and used as carriers to deliver an apoptosis-inducing plasmid encoding the tumor necrosis factor-related apoptosis-inducing ligand(pTRAIL)gene,along with the toll-like receptor(TLR7)agonist Rsiquimod(R848).The rationale behind this design is that pTRAIL can trigger cancer cell apoptosis by activating the DR4/5 receptor,while R848 can stimulate the immune microenvironment.Results:Experimental results demonstrated the synergistic effects of R848 and pTRAIL encapsulated by liposomes(RTL)in suppressing the proliferation of colorectal cancer cells.Moreover,further in vivo investigations revealed the strong anti-tumor efficacy of RTL in xenograft and orthotropic in situ models of colorectal cancer.Conclusions:These findings collectively highlight the therapeutic potential of R848/pTRAIL-loaded liposomes in the treatment of colorectal cancer.

Dissemination of Resistance Integrons and Genes Coding for Blse and Cabapenemases in the Urban Drainage Network in Cote d’Ivoire

Antibiotic resistance has become a major threat to human health worldwide. Environment, particularly the water environment, has long been overlooked as a player in the antibiotic resistance cycle, although its role remains unclear. These can provide an ideal setting for the acquisition and dissemination of antibiotic resistance, as they are frequently affected by anthropogenic activities. The objective of this study was to establish a diffusion map of resistance integrons used as genetic markers of resistance associated with antibiotic resistance conferring genes (ARGs). Total DNA extracts from non-cultivable bacterial communities were used for the analyses. These communities were obtained from wastewater samples from 14 sites upstream and downstream of drainage channels or effluents in the cities of Abidjan, Bouaké, and Yamoussoukro. The results obtained correspond to the number of positives among the treated samples (n = 39). Among the genetic markers of dissemination, class 1 integrons were the most evident in 94.8% of samples in Abidjan (93.3%), Bouaké (100%) and Yamoussoukro (91.6%). Class 2 integrons and class 3 integrons were found respectively in 41% and 51% of all samples. Genes coding for β-lactamases and blaTEM was identified in almost all samples at a rate of 97.4%. A co-presence of the three genes blaTEM, blaSHV and blaCTX-M is also remarkable in the sites of the city of Yamoussoukro. Among the genes coding for carbapenemases, only blaKPC 17.94%, blaNDM 30.76% and blaOXA48 38.46% were detected in the samples.

J-family genes redundantly regulate flowering time and increase yield in soybean

Soybean(Glycine max)is a short-day crop whose flowering time is regulated by photoperiod.The longjuvenile trait extends its vegetative phase and increases yield under short-day conditions.Natural variation in J,the major locus controlling this trait,modulates flowering time.We report that the three J-family genes influence soybean flowering time,with the triple mutant Guangzhou Mammoth-2 flowering late under short days by inhibiting transcription of E1-family genes.J-family genes offer promising allelic combinations for breeding.

Research on Developing an Assessment Scale for Tourism Experience Elements of Ancient Shu Road Heritage Trail

Cultural ancient roads,known in Chinese as gudao,serve as heritage trails that carry historical exchanges across various regions in China.Due to their extensive preservation,wide geographical distribution,diverse thematic variations,and considerable tourist appeal,these paths have emerged as representative heritage trails,increasingly transforming into a novel tourism product experience that is highly favored by tourists and recognized by government authorities.However,research on ancient roads for tourism in China currently lacks a systematic theoretical framework,as well as relevant policies,regulations,and standards to guide their practical development.Therefore,there is a pressing need to draw upon international best practices and conduct foundational research to develop an experience element system that aligns with the perceptions,behaviors,and consumption characteristics of Chinese tourists,thereby advancing theoretical exploration in this field.This study focuses on the representative Ancient Shu Road as a case study and employs a mixed-method approach that integrates qualitative and quantitative research.It aims to construct a tourist-centric scale for the experience elements of ancient road tourism while analyzing the interactive relationship between these experience elements and tourist needs.This study addresses a significant gap in the development of indicator systems for domestic studies of ancient road tourism experiences.Ultimately,the study establishes a comprehensive scale that encompasses three core categories—trail resources and environment,facilities and services,and modes of tourism activities—along with eight primary dimensions:core resources,surrounding cultural environment,surrounding natural environment,tourism reception facilities and services,infrastructure and support services,information facilities and information services,and outdoor and recreational activities.This scale consists of thirty-two specific items,providing a robust reference for future research endeavors.Additionally,the study proposes specific development strategies related to key mechanisms,spatial configuration,and facility construction to enhance the overall development of ancient road tourism.

Functional investigation and two-sample Mendelian randomization study of primary biliary cholangitis hub genes

BACKGROUND The identification of specific gene expression patterns is crucial for understanding the mechanisms underlying primary biliary cholangitis(PBC)and finding relevant biomarkers for diagnosis and therapeutic evaluation.AIM To determine PBC-associated hub genes and assess their clinical utility for disease prediction.METHODS PBC expression data were obtained from the Gene Expression Omnibus database.Overlapping genes from differential expression analysis and weighted gene coexpression network analysis(WGCNA)were identified as key genes for PBC.Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analyses were performed to explore the potential roles of key genes.Hub genes were identified in protein-protein interaction(PPI)networks using the Degree algorithm in Cytoscape software.The relationship between hub genes and immune cells was investigated.Finally,a Mendelian randomization study was conducted to determine the causal effects of hub genes on PBC.RESULTS We identified 71 overlapping key genes using differential expression analysis and WGCNA.These genes were primarily enriched in pathways related to cytokinecytokine receptor interaction,and Th1,Th2,and Th17 cell differentiation.We utilized Cytoscape software and identified five hub genes(CD247,IL10,CCL5,CCL3,and STAT3)in PPI networks.These hub genes showed a strong correlation with immune cell infiltration in PBC.However,inverse variance weighting analysis did not indicate the causal effects of hub genes on PBC risk.CONCLUSION Hub genes can potentially serve as valuable biomarkers for PBC prediction and treatment,thereby offering significant clinical utility.

Direct somatic embryogenesis and related gene expression networks in leaf explants of Hippeastrum ‘Bangkok Rose’

Hippeastrum, a highly diverse genus in the Amaryllidaceae family, is a valuable ornamental bulbous flowering plant. Somatic embryogenesis(SE) is an efficient method for mass production of Hippeastrum plantlets. Previous studies have been devoted to the in vitro propagation of Hippeastrum, but the SE and its regulatory networks are rarely reported. In this study, we established a direct SE method of Hippeastrum Bangkok Rose' using leaf bases as explants. MS supplemented with 1.00 mg·L^(-1)NAA +1.00 mg·L^(-1)KT + 0.25 mg·L^(-1)TDZ was the optimal medium for SE. Histological observations showed that the bipolar somatic embryo originated from the epidermal cell layer and underwent initiation,globular, scutellar and coleoptile stages. During SE, endogenous hormones of IAA, CTK, ABA, and SA were highly accumulated. Transcriptomic analysis revealed the genes encoding auxin biosynthesis/metabolic enzymes and efflux carriers were induced, while the auxin receptor of TIR1 and ARF transcriptional repressor of Aux/IAA were down-regulated and up-regulated, respectively, leading to suppression of auxin signaling. In contrast, cytokine signaling was promoted at the early stage of SE, as biosynthesis, transport, and signaling components were up-regulated.Various stress-related genes were up-regulated at the early or late stages of SE. Chromatin remodeling could also be dynamically regulated via distinct expression enzymes that control histone methylation and acetylation during SE. Moreover, key SE regulators, including WOXs and SERKs were highly expressed along with SE. Overall, the present study provides insights into the SE regulatory mechanisms of the Hippeastrum.