Objective:To screen for microRNAs(miRNAs)highly expressed in the serum exosomes(Exo)of esophageal squamous cell carcinoma(ESCC)patients and analyze their relationship with the clinicopathological characteristics of the patients,and to explore the potential of Exo-derived miRNAs as clinical auxiliary diagnostic markers for ESCC.Methods:Serum and relevant clinical data of 50 healthy subjects and 45 newly diagnosed ESCC patients admitted to the Fourth Hospital of Hebei Medical University between December 2021 and June 2023 were collected,serving as the control group and the ESCC group respectively.The Gene Expression Omnibus(GEO)database and qPCR were used to screen and identify the candidate miRNA for increased expression in the serum of ESCC patients-miR-1246.The diagnostic efficacy of serum miR-1246 for ESCC was analyzed by the receiver operating characteristic curve.The relationship between miR-1246 and the clinical feature progression of ESCC patients was analyzed by Logistic regression,and the relationship between miR-1246 and the clinicopathological characteristics of ESCC patients was analyzed by the χ2 test.Exosomes in the serum of the subjects were isolated,purified and characterized for verification.The expression of miR-1246 in Exo was detected by qPCR.ESCC KYSE150 and KYSE30 cells were routinely cultured.mimics-NC and miR-1246 mimics were transfected respectively into KYSE150 cells using Lipofectamine 2000.Inhibitor-NC and miR-1246 inhibitor were transfected into KYSE30 cells,which were respectively denoted as the minics-NC,miR-1246 mimics,inhibitor-NC and miR-1246-inhibitor groups.KYSE150 and KYSE30 cells were treated with Exo derived from KYSE150 cells in the mimics-NC and miR-1246 mimics groups.The proliferation,migration and invasion abilities of cells in each group were detected by the CCK-8 assay,scratch wound healing assay and Transwell chamber assay respectively.The expressions of Exo markers,epithelial-mesenchymal transition-related proteins,TET family methylcytosine dioxygenase 2(TET2)and cell adhesion molecule 1(CADM1)proteins in each group of cells were detected by WB assay.The targeting binding relationship between miR-1246 and TET2 and CADM1 was verified by the dual-luciferase reporter gene assay.Results:Bioinformatics screening showed that the miRNA with the most significant differential expression in the serum of ESCC patients was miR-1246.The serum Exo extracted from the patients conformed to the typical Exo characteristics.The expression level of serum Exo-miR-1246 in ESCC patients at stages Ⅰ-Ⅱ was significantly higher than that in healthy subjects(P<0.01);the level of serum Exo-miR-1246 in ESCC patients at stages Ⅲ-Ⅳ was significantly higher than that in patients at stages Ⅰ-Ⅱ(P<0.01).ROC curve analysis showed that Exo-miR-1246 in serum had a high value for auxiliary differential diagnosis of ESCC(P<0.05),and the auxiliary diagnostic efficacy of Exo-miR-1246 for the clinical progression of ESCC patients was higher than that of CEA and SCC-Ag(P<0.05).The combined detection of the three could further improve the efficacy of auxiliary diagnosis of patient staging(P<0.01).Exo-miR-1246 might be an independent risk factor for the clinical progression of ESCC patients(P<0.05).The expression level of serum Exo-miR-1246 was associated with the T-stage,N-stage and clinical stage of ESCC(P<0.01).Overexpression of miR-1246 could promote the proliferation,migration,invasion,epithelial-mesenchymal transition and inhibit apoptosis of ESCC cells,while inhibition of miR-1246 had the opposite effect.Database data analysis found that TET2 and CADM1 were the target genes of miR-1246.The dual-luciferase reporter gene assay confirmed that miR-1246 could directly bind to TET2 and CADM1 mRNA and inhibit their expressions(P<0.01).Treatment of KYSE150 and KYSE30 cells with Exo derived from cells overexpressing miR-1246 had the same effect as overexpressing miR-1246 in these cells.Conclusion:Exo-derived miR-1246 has the potential to be a clinical auxiliary diagnostic marker for ESCC.It may affect the occurrence and development of ESCC by regulating the expression levels of TET2 and CADM1.

Objective:To screen for microRNAs(miRNAs)highly expressed in the serum exosomes(Exo)of esophageal squamous cell carcinoma(ESCC)patients and analyze their relationship with the clinicopathological characteristics of the patients,and to explore the potential of Exo-derived miRNAs as clinical auxiliary diagnostic markers for ESCC.Methods:Serum and relevant clinical data of 50 healthy subjects and 45 newly diagnosed ESCC patients admitted to the Fourth Hospital of Hebei Medical University between December 2021 and June 2023 were collected,serving as the control group and the ESCC group respectively.The Gene Expression Omnibus(GEO)database and qPCR were used to screen and identify the candidate miRNA for increased expression in the serum of ESCC patients-miR-1246.The diagnostic efficacy of serum miR-1246 for ESCC was analyzed by the receiver operating characteristic curve.The relationship between miR-1246 and the clinical feature progression of ESCC patients was analyzed by Logistic regression,and the relationship between miR-1246 and the clinicopathological characteristics of ESCC patients was analyzed by the χ2 test.Exosomes in the serum of the subjects were isolated,purified and characterized for verification.The expression of miR-1246 in Exo was detected by qPCR.ESCC KYSE150 and KYSE30 cells were routinely cultured.mimics-NC and miR-1246 mimics were transfected respectively into KYSE150 cells using Lipofectamine 2000.Inhibitor-NC and miR-1246 inhibitor were transfected into KYSE30 cells,which were respectively denoted as the minics-NC,miR-1246 mimics,inhibitor-NC and miR-1246-inhibitor groups.KYSE150 and KYSE30 cells were treated with Exo derived from KYSE150 cells in the mimics-NC and miR-1246 mimics groups.The proliferation,migration and invasion abilities of cells in each group were detected by the CCK-8 assay,scratch wound healing assay and Transwell chamber assay respectively.The expressions of Exo markers,epithelial-mesenchymal transition-related proteins,TET family methylcytosine dioxygenase 2(TET2)and cell adhesion molecule 1(CADM1)proteins in each group of cells were detected by WB assay.The targeting binding relationship between miR-1246 and TET2 and CADM1 was verified by the dual-luciferase reporter gene assay.Results:Bioinformatics screening showed that the miRNA with the most significant differential expression in the serum of ESCC patients was miR-1246.The serum Exo extracted from the patients conformed to the typical Exo characteristics.The expression level of serum Exo-miR-1246 in ESCC patients at stages Ⅰ-Ⅱ was significantly higher than that in healthy subjects(P<0.01);the level of serum Exo-miR-1246 in ESCC patients at stages Ⅲ-Ⅳ was significantly higher than that in patients at stages Ⅰ-Ⅱ(P<0.01).ROC curve analysis showed that Exo-miR-1246 in serum had a high value for auxiliary differential diagnosis of ESCC(P<0.05),and the auxiliary diagnostic efficacy of Exo-miR-1246 for the clinical progression of ESCC patients was higher than that of CEA and SCC-Ag(P<0.05).The combined detection of the three could further improve the efficacy of auxiliary diagnosis of patient staging(P<0.01).Exo-miR-1246 might be an independent risk factor for the clinical progression of ESCC patients(P<0.05).The expression level of serum Exo-miR-1246 was associated with the T-stage,N-stage and clinical stage of ESCC(P<0.01).Overexpression of miR-1246 could promote the proliferation,migration,invasion,epithelial-mesenchymal transition and inhibit apoptosis of ESCC cells,while inhibition of miR-1246 had the opposite effect.Database data analysis found that TET2 and CADM1 were the target genes of miR-1246.The dual-luciferase reporter gene assay confirmed that miR-1246 could directly bind to TET2 and CADM1 mRNA and inhibit their expressions(P<0.01).Treatment of KYSE150 and KYSE30 cells with Exo derived from cells overexpressing miR-1246 had the same effect as overexpressing miR-1246 in these cells.Conclusion:Exo-derived miR-1246 has the potential to be a clinical auxiliary diagnostic marker for ESCC.It may affect the occurrence and development of ESCC by regulating the expression levels of TET2 and CADM1.

[摘 要] 目的：检测食管鳞状细胞癌（ESCC）患者血清中高迁移率族蛋白B1（HMGB1）和吲哚胺-2，3-双加氧酶（IDO）的表达水平并探讨两者与临床病理特征及淋巴细胞亚群的相关性。方法：选取2021年3月至2022年8月在河北医科大学第四医院初次住院治疗的95例ESCC患者作为ESCC组，另选取40例健康体检人群作为对照组。ELISA法检测全部研究对象的血清HMGB1和IDO水平及不同组ESCC细胞培养上清中HMGB1、IDO和p65水平，流式细胞术检测全部研究对象外周血淋巴细胞亚群水平。WB法检测仅敲低HMGB1基因表达或敲低HMGB1后再加入NF-κB信号通路激活剂对ESCC细胞HMGB1、IDO和p65表达的影响。结果：ESCC组患者血清HMGB1和IDO水平明显高于对照组（均P<0.01）；血清HMGB1和IDO表达水平升高是ESCC临床进展的独立危险因素（均P<0.01），二者联合检测对ESCC临床进展预测价值更高（P<0.01）；血清HMGB1和IDO与ESCC患者的T分期、N分期和临床分期有明显关联（均P<0.05）； ESCC组患者血清HMGB1与外周血CD3+ T细胞、CD4+ T细胞、B细胞和NK细胞绝对计数值呈显著负相关，而与Treg细胞百分率呈显著正相关（均P<0.05），血清IDO与外周血CD3+ T细胞百分率和绝对计数值、CD4+ T细胞百分率和绝对计数值、CD8+ T细胞和B细胞绝对计数值呈显著负相关，而与Treg细胞百分率呈显著正相关（均P<0.05）；血清HMGB1和IDO表达水平呈显著正相关（P<0.01）。si-HMGB1组KYSE30和ECA109细胞及其培养上清液中IDO和p65表达水平明显低于si-NC组和si-HMGB1+PMA组（均P<0.05）。结论：血清HMGB1和IDO与ESCC临床进展和机体免疫功能密切相关，具有成为ESCC肿瘤标志物和免疫治疗新靶点的潜力。HMGB1可能通过NF-κB信号通路促进IDO表达，进行双靶点联合治疗可能会取得更好的疗效。

Alzheimer's disease (AD) is the first degenerative disease of the nervous system, but no drugs have been found to reverse AD progression. Starting from 2 major pathogenesis of AD, namely amyloid beta (Aβ) cascade and Tau protein, this study systematically reviews anti-Aβ or Tau protein AD new drugs that have entered clinical research; this study also expounds their clinical trial findings and mechanisms, and analyzes the reasons for their success or failure to provide a theoretical basis for AD drug exploitation.

Objective:To assess the differences of the 2016 and 2009 edtion guidelines on diastolic dysfunction in sepsis patients.Methods:A single-center, prospective study was conducted. The relevant information of sepsis patients in Intensive Care Unit (ICU) were analyzed from October 2016 to January 2019. Patients’ transthoracic echocardiography at the first 24 h and 3rd day of their admission and left ventricular diastolic dysfunction were stratified according to the 2009 and 2016 edition guidelines. Patients’ characteristics, arterial blood gas analysis, and blood biochemical indexes were recorded at the first 24 h of the ICU admission. Additionally, the following information were retrieved during ICU stay: site of infection, frequency of adrenaline and dobutamine, maximal dose of norepinephrine, use of hydrocortisone, invasive mechanical ventilation and renal replacement therapy. The rank-sum test of two independent samples was used to compare the differences in the diagnosis of left ventricular diastolic dysfunction.Results:A total of 196 patients with sepsis or septic shock were screened, and 86 patients were excluded. Finally, clinical data of 110 patients were included in the analysis.The median time of the first ultrasound examination in ICU was 17 h. Among the patients with different diastolic function severity in baseline data analysis, only age was significantly different. According to the 2016 edition guidelines, 43 (39%) of 110 patients had diastolic dysfunction and another 30 (27%) had indeterminate diastolic dysfunction within 24 h of ICU admission. According to the 2009 edition guidelines, 40 (36%) patients had diastolic dysfunction and 58 (53%) patients had indeterminate diastolic dysfunction. The diagnosis of left ventricular diastolic dysfunction of different grades was significantly different between the 2016 and 2009 edition guidelines ( Z=4.92, P<0.01). According to the 2016 edition guidelines at the 3rd day of ICU admission, 52 (47%) patients were diagnosed with diastolic dysfunction and 18 (18%) were diagnosed with indeterminate diastolic dysfunction; According to the 2009 edition guidelines, 50 (46%) of these patients were diagnosed with diastolic dysfunction and 45 (41%) had indeterminate diastolic function. Similarly, there was a large difference in diagnosis ( Z=4.60, P<0.01). Subgroup analysis of patients with normal systolic function (ejection fraction > 50%) showed that the diagnosis of left ventricular diastolic dysfunction were significantly different at the first 24 h and the 3rd day of ICU admission ( Z=4.34, P<0.01 and Z=5.71, P<0.01). Conclusions:The 2016 edition guidelines identify a significantly higher incidence of dysfunction in patients with severe sepsis and septic shock compared to the 2009 edition guidelines. Although the 2016 edition guidelines seem to be an improvement, issues remain with the application of guidelines using traditional measures of diastolic dysfunction in this cohort.

The main constituents of a typical medicinal herb,Polygonum muMflorum (Heshouwu in Chinese),that induces idiosyncratic liver injury remain unclear.Our previous work has shown that cotreatment with a nontoxic dose of lipopolysaccharide (LPS) and therapeutic dose of Heshouwu can induce liver injury in rats,whereas the solo treatment cannot induce observable injury.In the present work,using the constituent "knock-out" and "knock-in" strategy,we found that the ethyl acetate (EA) extract of Heshouwu displayed comparable idiosyncratic hepatotoxicity to the whole extract in LPS-treated rats.Results indicated a significant elevation of plasma alanine aminotransferase,aspartate aminotransferase,and liver histologic changes,whereas other separated fractions failed to induce liver injury.The mixture of EA extract with other separated fractions induced comparable idiosyncratic hepatotoxicity to the whole extract in LPS-treated rats.Chemical analysis further revealed that 2,3,5,4'-tetrahydroxy trans-stilbene-2-O-β-glucoside (trans-SG) and its cis-isomer were the two major compounds in EA extract.Furthermore,the isolated cis-,and not its trans-isomer,displayed comparable idiosyncratic hepatotoxicity to EA extract in LPS-treated rats.Higher contents of cis-SG were detected in Heshouwu liquor or preparations from actual liver intoxication patients associated with Heshouwu compared with general collected samples.In addition,plasma metabolomics analysis showed that cis-SG-disturbing enriched pathways remarkably differed from trans-SG ones in LPS-treated rats.All these results suggested that cis-SG was closely associated with the idiosyncratic hepatotoxicity of Heshouwu.Considering that the cis-trans isomerization of transSG was mediated by ultraviolet light or sunlight,our findings serve as reference for controlling photoisomerization in drug discovery and for the clinical use of Heshouwu and stilbene-related medications.

The study was conducted to investigate the role of vitamin E in the high altitude hypoxia-induced damage to the intestinal barrier in rats. Sprague-Dawley rats were divided into control (Control), high altitude hypoxia (HH), and high altitude hypoxia+vitamin E (250 mg/kg BW*d) (HV) groups. After the third day, the HH and HV groups were placed in a hypobaric chamber at a stimulated elevation of 7000 m for 5 days. The rats in the HV group were given vitamin E by gavage daily for 8 days. The other rats were given equal volume saline. The results showed that high altitude hypoxia caused the enlargement of heart, liver, lung and kidney, and intestinal villi damage. Supplementation with vitamin E significantly alleviated hypoxia-caused damage to the main organs including intestine, increased the serum superoxide dismutase (SOD) (p< 0.05), diamino oxidase (DAO) (p< 0.01) levels, and decreased the serum levels of interleukin-2 (IL-2) (p< 0.01), interleukin-4 (IL-4) (p<0.001), interferon-gamma (IFN-gamma) (p<0.01) and malondialdehyde (MDA) (p<0.001), and decreased the serum erythropoietin (EPO) activity (p<0.05). Administration of vitamin E significantly increased the S-IgA (p<0.001) in ileum and significantly improved the expression levels of occludin and IkappaBalpha, and decreased the expression levels of hypoxia-inducible factor 1 alpha and 2 alpha (HIF-1alpha and HIF-2alpha), Toll-like receptors (TLR4), P-IkappaBalpha and nuclear factor-kappaB p65(NF-kappaB P65) in ileum compared to the HH group. This study suggested that vitamin E protectis from intestinal injury caused by high altitude hypoxia environment. These effects may be related to the HIF and TLR4/NF-kappaB signaling pathway.
Altitude* ; Animals ; Anoxia* ; Erythropoietin ; Heart ; Hypoxia-Inducible Factor 1 ; Ileum ; Interferon-gamma ; Interleukin-2 ; Interleukin-4 ; Intestinal Mucosa ; Intestines ; Kidney ; Liver ; Lung ; Malondialdehyde ; Occludin ; Oxidoreductases ; Rats* ; Rats, Sprague-Dawley ; Superoxide Dismutase ; Toll-Like Receptors ; Vitamin E* ; Vitamins*

To investigate chemical constituents contained in cytotoxic petroleum ether extractive fractions from ethanol extracts of Cirsium setosum. The constituents were separated and purified by a combination of various chromatographic methods including silica gel, Sephadex LH-20, and preparative HPLC. Structures of the isolates were elucidated by spectroscopic methods including 1D, 2D NMR and MS methods. The compound structures were also determined by reference to literature. Twelve compounds were separated from the petroleum ether fraction of ethanolic extract and elucidated as lupenyl acetate (1), lupeol (2), lupenone (3), beta-amyrin (4), psi-taraxasterol (5), psi-taraxasteryl acetate (6), taraxasteryl acetate (7), marsformoxide B (8), alpha-amyrenone (9), beta-amyrenone (10), taraxasterone (11) and psi-taraxasterone (12). Of them, compounds 3, 5, 7-12 were separated from this genus for the first time.
Chromatography, High Pressure Liquid ; Cirsium ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; Oleanolic Acid ; analogs & derivatives ; chemistry ; Pentacyclic Triterpenes ; chemistry ; Sterols ; chemistry ; Triterpenes ; chemistry