BACKGROUND:Tumor-associated macrophages (TAMs) are crucial in hepatocellular carcinoma (HCC) progression and prognosis, making them promising immunotherapy targets. In traditional Chinese medicine (TCM), qi stagnation and blood stasis are linked to the HCC tumor microenvironment (TME), but few studies explore the effects of related TCM herbs on the TME. Calceolarioside B, a key phenylethanoid glycoside in Akebiae Fructus, has not been well studied for its pharmacological activities or molecular targets, and its role in HCC remains unclear. PURPOSE:This study aimed to investigate the effects of Calceolarioside B on TAMs in HCC and clarify its potential targets and regulatory mechanisms. METHODS:Murine intrahepatic transplantation HCC models and macrophage-HCC cell co-culture systems were used to investigate the effects of Calceolarioside B on M2-like TAMs polarization and infiltration, and tumor growth. Cellular thermal shift assay, small molecular pull-down assay and surface plasmon resonance were utilized to identify the potential targets regulating M2-like TAMs. Single-cell RNA sequencing and TCGA dataset analyses clarified the differential expression, prognosis, and TAMs association of the potential targets in HCC. RESULTS:Calceolarioside B reduces M2-like TAMs polarization and infiltration in the TME by binding to and inhibiting matrix metallopeptidase-12 (MMP12) form both macrophages and HCC cells, thereby preventing immunosuppressive effects. Public database analysis revealed that MMP12 overexpression promoted macrophage infiltration, with MMP12+ macrophages preferentially aggregating in primary and metastatic HCC tumors. CONCLUSION:Calceolarioside B is identified as a novel MMP12 inhibitor modulating TAMs in the TME, offering a potential TAM-targeting strategy for HCC therapy.

The toxicity and side effects of chemotherapeutic drugs remain a crucial obstacle to the clinical treatment of hepatocellular carcinoma (HCC). Identifying combination therapy from Chinese herbs to enhance the sensitivity of tumors to chemotherapeutic drugs is of particular interest. Astragalus polysaccharide (APS), one of the natural active components in Astragalus membranaceus, has been reported to exhibit anti-tumor properties in diverse cancer cell lines. The aim of this study was to determine the effect of APS on Doxorubicin (Dox)-induced apoptosis in HCC and the underlying mechanism. The results showed that APS dose-dependently promoted Dox-induced apoptosis and enhanced endoplasmic reticulum (ER) stress. Additionally, APS decreased the mRNA level and protein stability of O-GlcNAc transferase (OGT), and increased the O-GlcNAcase (OGA) expression. Furthermore, OGT lentiviral transfection or PugNAc (OGA inhibitor) treatment reversed the ER stress and apoptosis induced by the combination of Dox and APS. A xenograft tumor mouse model confirmed that the combination of APS and Dox showed an advantage in inhibiting tumor growth in vivo. These findings suggested that APS promoted Dox-induced apoptosis in HCC cells through reducing the O-GlcNAcylation, which led to the exacerbation of ER stress and activation of apoptotic pathways.

ObjectiveTo study the pathway of cis-dichlorodiamineplatinum (DDP) inhibiting the synthesis of steroid hormones in mice, and to observe the intervention effect of dehydroepiandrosterone (DHEA). MethodsSixty adult ICR mice were randomly divided into three groups: control group, DDP modeling group, and DHEA group, with 10 male and 10 female mice in each group. The DDP modeling group mice were intraperitoneally injected with DDP solution at a dose of 2.5 mg·kg-1·d-1, once every 3 days, a total of 7 times. On the same day of modeling, the control group mice were injected with an equal amount of physiological saline intraperitoneally. The DEHA treatment group mice were treated with DDP and given a dose of 8.3 mg·kg-1·d -1 of DHEA by gavage for 21 consecutive days. The changes of fatigue indexes of mice were observed by open field, grip and rod rotation tests. The morphology changes of adrenal gland, testicular and ovarian tissue were observed by pathological section and HE staining. The levels of serum steroid hormones were detected by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The mRNA and protein expression levels of the related genes of the hypothalamus, hypophysis, adrenal, testis and ovary were tested by real-time fluorescent quantitative PCR (RT-qPCR) and Western blotting. ResultsCompared with control group, both male and female mice in DDP modeling group were significantly losing weight (P＜0.05), their abilities in horizontal movement and vertical movement decreased (all P＜0.05), and the stay time and grip also significantly decreased (all P＜0.05) in female mice. Indexes of fatigue were improved after DHEA supplement (all P＜0.05). In the DDP modeling group, the arrangement of spermatogenic cells at all levels in the testicular tissue was disordered and the testicular interstitial edema was observed, and a large number of primordial follicles in the ovarian tissue were activated, the number of atresia follicles increased, and the number of granulosa cells in the follicles decreased; while in the DHEA group, the damaged phenotype of testicles and ovaries was significantly improved. Compared with control group, the levels of serum testosterone and dihydrotestosterone in both male and female DDP modeling mice significantly decreased (P＜0.01), the pregnenolone was down-regulated but corticosterone was up-regulated significantly (P＜0.05) in male mice, the corticosterone was down-regulated significantly (P＜0.05) in female mice. Compared with the DDP group, after DHEA supplement, the pregnenolone in male mice and the progesterone in female mice increased significantly (P＜0.05), but the pregnenolone in female mice and the progesterone in male mice decreased significantly (P＜0.05). Compared with control group, the expression levels of Cyp21a1 and Cyp11a1 genes in the adrenal gland and Gnrh gene in the hypothalamus of male and female mice in the DDP modeling group significantly decreased (all P＜0.05); the expression levels of Hsd3b2 gene in the adrenal gland, Star, Cyp11a1, and Lhr genes in the ovaries, Crh, Pomc, and Lhb genes in the hypothalamus, pituitary, and pituitary of female mice significantly decreased (all P＜0.05); the expression levels of Star gene and StAR protein in the testicles of male mice, as well as Fshb and Lhb genes in the pituitary gland, were significantly down-regulated (all P＜0.05). After DHEA supplement, compared with the DDP modeling group, the mRNA expression levels of Cyp17a1 in the adrenal gland of male mice and Cyp17a1, Lhr and Fshr genes in testis were down-regulated significantly (P＜0.05); the expression level of Cyp11a1 gene in the adrenal gland of female mice was also decreased (P＜0.05); while the expression levels of Hsd3b2 gene in the adrenal gland, Star, Cyp11a1, Hsd3b2 and Lhr gene in the ovary, and Lhb gene in the pituitary gland were all up-regulated ( P＜0.05). ConclusionThe function of hypothalamus-pituitary-adrenal/gonadal axis was inhibited by DDP intermittent injection, especially in female. Supplementation of DHEA can help regulate the homeostasis of steroid hormone levels.

目的 研究顺铂诱发药源性虚证性小鼠类固醇激素合成功能的改变.方法 72只雄性ICR小鼠分为对照组32只和顺铂组40只,顺铂组予10mg/kg顺铂腹腔注射,每周1次,共5次(d1、d8、d15、d22、d29).每次造模后1周各组取8只检测相应指标,以ELISA检测睾酮和皮质酮含量、RT-qPCR检测下丘脑、垂体、肾上腺和睾丸类固醇激素合成酶相关基因表达.Western Blot 检测 肾上腺 StAR、CYP11A1、CYP21A2、CYP11B1;睾丸 StAR、CYP11A1、CYP17A1、HSD3B2 蛋白表达.结果 ①顺铂给药4次(d28)后显著下降小鼠体重,胸腺与肾脏萎缩,脾肿大.②小鼠血清皮质酮在顺铂给药2次(d14)及4次(d28)显著上升,末次(d30)显著下降.血清睾酮含量在给药1次后(d7)明显上升,给药4次和末次(d28、d30)明显下降.③随顺铂累积肾上腺皮质束状带明亮细胞增多.④随顺铂累积,曲细精管改变、生精细胞紊乱加重,给药4次(d28)后曲细精管内改变显著,支持细胞减少,间质增厚,间质细胞水肿明显.⑤顺铂组肾上腺在给药1次(d7)后Cyp21a1基因表达和CYP11A1蛋白表达上调,但Cyp11b2基因表达下调;给药2次(d14)后Star、Cyp11a1、Cyp11b1、Cyp11b2和Cyp21a1基因表达均下调;给药3次(d21)Cyp11a1和Cyp21a1基因表达下调;给药4次(d28)Cyp11b1基因表达上调.⑥顺铂组睾丸在给药2次(d14)后Star、Cyp11a1、Cyp17a1、Hsd17b3和Ar基因表达均下调;给药3次(d21)Hsd3b2和Ar基因表达上调;给药4次(d28)Star、Cyp11a1、Cyp17a1、Hsd3b2、Hsd17b3和Ar基因表达均显著上调,StAR蛋白表达上调;末次给药(d30)Star、Cyp11a1、Cyp17a1、Hsd3b2、Hsd17b3和Ar基因表达均上调,StAR、CYP11A1 和 CYP17A1蛋白表达显著增强.⑦顺铂组下丘脑在给药2次(d14)GnRH基因表达增强,给药1次、4次和末次(d7、d28、d30)CRH基因表达均增强.⑧顺铂组垂体在给药1次(d7)Pou1f1基因表达上调而Fshb和Lhb基因表达均下调;给药4次(d28)Pomc基因表达上调.结论 顺铂可损害类固醇激素合成的肾上腺皮质轴与睾丸轴,尤其对睾丸各细胞的累积性损害可能是诱发肾精不足证的主要机制.

木通属Akebia Decne.目前发现5种,其植物在中医药治疗上应用广泛,药用品种主要是木通(五叶木通)Akebia quinate、三叶木通A.trifoliata和白木通A.trifoliate subsp.australis;主要药用部位为其藤茎和果实,具有利尿通淋和疏肝理气等功效.近年来木通属植物也常用于中医药的肿瘤防治,许多研究表明其提取物对胃癌、肝癌等肿瘤细胞的增殖产生抑制作用,而来自木通属植物不同部位提取物的抗癌作用也不尽相同,其抗肿瘤机制包括诱导凋亡、抑制侵袭与转移、阻滞细胞周期、抑制肿瘤细胞血管生成等.对木通属植物的藤茎、果实、种子提取物和其天然产物的抗肿瘤作用研究进行归纳综述,为木通属植物在中医药肿瘤防治方面的应用提供新思路.

Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease around the world. However, no specific medicine has been approved for NAFLD treatment. Our study was conducted to explore the role and mechanism of TRIM59 in NAFLD, aiming to provide a novel target for NAFLD treatment. Here, the expression of TRIM family members was detected in 10 mild and severe NAFLD tissues as well as 10 normal tissues. TRIM59 expression was verified in 10 normal tissues and 25 mild and severe NAFLD tissues. Palmitic acid and high-fatty diet were used for the construction of NAFLD models. Oil Red O staining was used to detect the level of steatosis. The content of TNF-α, IL-6, and IL-8 was measured to reflect the level of inflammation. Lipid reactive oxygen species was estimated by flow cytometry. We found that TRIM59 was highly expressed in NAFLD tissues compared with normal liver tissues. The inhibition of TRIM59 could inhibit the steatosis and inflammation in NAFLD, whereas its overexpression exhibited reversed effects. The application of ferroptosis inhibitor, deferoxamine, could markedly ameliorate steatosis and inflammation, which was mediated by overexpressed TRIM59. Besides, TRIM59 was demonstrated to interact with GPX4 and promoted its ubiquitination. The overexpression of GPX4 could significantly reverse the pathogenic effects of TRIM59 in NAFLD. Additionally, the inhibition of TRIM59 appeared to be a promising strategy to ameliorate NAFLD in mice model. In summary, our study revealed that TRIM59 could promote steatosis and ferroptosis in NAFLD via enhancing GPX4 ubiquitination. TRIM59 could be a potential target for NAFLD treatment.

A two-dimensional (2D) glycomaterial for targeted delivery of maytansine to liver cancer cells was developed. Host-guest interaction between a galactosyl dye and human serum albumin (HSA) produces supramolecular galactoside-HSA conjugates, which are then used to coat 2D MoS2. The 2D glycomaterial was shown to be capable of the targeted delivery of maytansine to a liver cancer cell line that highly expresses a galactose receptor, resulting in greater cytotoxicity than maytansine alone.

Metastasis remains a crucial obstacle to the clinical treatment of hepatocellular carcinoma (HCC). Investigating the potential anti-tumor compounds from medicinal herb against HCC metastasis is of particular interest. As a triterpenoid saponin, α-Hederin has been reported to exhibit cytotoxicity for diverse cancer cell lines by inducing mitochondrial related apoptosis or autophagic cell death. Nevertheless, little is known about the inhibitory effect of α-Hederin on the metastasis of HCC and its underlying mechanisms. Here, we integrated well-established target prediction webtool and molecular docking methods to predict the potential targets for α-Hederin, and finally focused on PTAFR, the receptor for platelet-activating factor (PAF). Activation of PAF/PTAFR pathways has been reported to be contribution to the initiation and progression of cancer. We showed for the first time that non-cytotoxic concentration of α-Hederin inhibited cell migration and invasion induced by PAF in HCC cells, as well as lung metastasis in vivo. Moreover, we demonstrated α-Hederin reduced the PAF-induced matrix metalloproteinase-2 expression through inhibiting the activation of STAT3 in PAF stimulated HCC cells. These findings suggest that α-Hederin functions as a prospective inhibitor of PTAFR and may be utilized as an optional candidate for treatment of HCC.

天癸是《黄帝内经》认识人类生育能力的物质,基于男性与女性的差异,提出了天癸兴衰在女子表现为"七七理论"、在男子表现为"八八理论",且天癸兴衰理论对临床具有重要的指导价值.传统中医理论对天癸的认识以定性表述为主,而现代医学善于以量化方式精确认识人类生命现象.基于此,以定性与定量相结合的方式,阐述全生命周期中天癸的相关物质动态变化,并认为:①女子卵巢、男子睾丸及肾上腺皮质是天癸发生的重要源泉,女子雌激素与孕酮、男子脱氢表雄酮与睾酮能够较客观地反映全生命周期中天癸的物质基础;②性激素在女性"七七"阶段急骤下降,而在男性"八八"阶段仅为渐进式下降,因而临床中女性围绝经期综合征特征显著,而男性几乎不表现出相应的综合征;③肾气是调控天癸发生的主要因素,类似于人体高级中枢大脑皮质、下丘脑、垂体功能;④以现代科学术语表达与解析传统中医理论概念将有助于初学者更客观、理性地认识中医天癸的学术内涵.

OBJECTIVE:To investigate the efficacy of Yuzhizi seed extract (FAQSE) on inhibiting the proliferation of hepatocellular carcinoma (HCC) cells in vitro and to explore the anti-HCC action mechanism of FAQSE.METHODS:Human HCC HepG2 and Huh7 cells were used to investigate the anti-HCC effect of FAQSE. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) method was used to measure cell viability. Affymetrix microarray was adopted to detect the expression of transcriptome. The differentially expressed genes (DEGs) of each cell line were identified. For co-DEGs of both cell lines, the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were enriched using the Database for Annotation, Visualization and Integrated Discovery (DAVID), and the network analysis of protein-protein interaction (PPI) was mapped using the Retrieval of Interacting Genes/Proteins (STRING) and Cytoscape software. Some important genes in the PPI network of co-DEGs were selected to verify by quantitative real-time reverse transcription-polymerase chain reaction, Western blot and enzyme-linked immunosorbent assay.RESULTS:FAQSE decreased the viability of HepG2 and Huh7 cells. There were 211 co-upregulated and 86 co-downregualted genes in both cell lines after FAQSE treatment. The enriched GO terms of co-upregulated DEGs were primarily involved cell-cell adhesion, viral process, transcription initiation from RNA polymerase II promoter, positive regulation of transcription from RNA polymerase II promoter and actin cytoskeleton organization. The GO terms of co-downregulated DEGs were mainly enriched in the processes of SRP-dependent cotranslational protein targeting to membrane, viral transcription, nuclear-transcribed mRNA catabolic process, nonsense-mediated decay, translational initiation and rRNA processing. Main KEGG pathways of co-upregulated DEGs were endocytosis, glutathione metabolism, protein processing in endoplasmic reticulum, synaptic vesicle cycle and lysosome. The major KEGG pathways of co-downregulated DEGs were ribosome, biosynthesis of amino acids, arginine and proline metabolism, systemic lupus erythematosus and complement and coagulation cascades. The top 10 co-DEGs with high hub nodes in STRING analysis were ribosomal protein S27a, transferrin, ribosomal protein S20, ribosomal protein L9, protein phosphatase 2 regulatory subunit B alpha, transthyretin, thioredoxin reductase 1, ribosomal protein L3, ribophorin I and ribosomal protein L24. Alpha-fetoprotein (AFP) was also co-downregulated and contained in the PPI network. The mRNA and protein expression of most verified genes was consistent with the results of co-DEGs analysis. And the AFP level was significantly reduced after FAQSE treatment.CONCLUSIONS:A series of genes and pathways of HepG2 and Huh7 cells were changed after FAQSE treatment, which might be the targets of FAQSE against HCC and worthy of further study. AFP might be important one of them.

本文介绍方肇勤在中医辨证论治理论方面的学术成果.从辨证论治理论研究及其计算机程序化模拟研究、辨证论治动物实验方法学研究、肝癌同病异证证候特征及其物质基础研究、中药复方及其拆方后不同治法的研究予以简要梳理.方肇勤在学术上的突出成就体现在辨证中提出了"新值"和"潜值""显值"等概念,创建了大鼠/小鼠辨证论治实验方法学,率先提出与践行按照不同治法拆方的思路研究复方药效,领先开展大规模临床病例的证候流行病学调查研究,同步从神经内分泌免疫组织基因表达谱全方位探索证候物质基础,所取得的研究成果丰富与发展了当代中医基础理论.

目的 研究"补气生津"人参-黄芪与"填精补肾"熟地-山茱萸对皮质激素生成的影响.方法 采用0.25-8 g·L-1人参-黄芪与0.25-8 g·L-1熟地-山茱萸水煎醇沉液分别干预小鼠肾上腺皮质瘤细胞(Y1 adrenocortical cell,Y1细胞)48 h,采用MTT比色法(MTT assay,MTT)检测细胞增殖,运用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测细胞分泌皮质酮含量,实时荧光定量PCR(Real-time Quantitative polymerase chain reaction,qPCR)检测类固醇激素合成酶基因表达,免疫蛋白印迹法(Western blot,WB)检测蛋白表达.结果 与空白组(Control,Con)比较,0.25-8 g·L-1人参-黄芪和熟地-山茱萸均抑制Y1细胞增殖(P<0.01);4 g·L-1和8 g·L-1人参-黄芪促进皮质酮分泌(P<0.01),4 g·L-1和8 g·L-1熟地-山茱萸抑制皮质酮分泌(P<0.01);8 g·L-1人参-黄芪增强CYP11A1、CYP21A2、CYP11B1蛋白表达(P<0.05),8 g·L-1熟地-山茱萸抑制CYP11A1、CYP21A2、CYP11B1蛋白表达(P<0.01);8 g·L-1人参-黄芪抑制Star基因表达(P<0.05),而8 g·L-1熟地-山茱萸促进Star基因表达(P<0.01),4 g·L-1与8 g·L-1人参-黄芪和0.25-8 g·L-1熟地-山茱萸均能显著抑制Cyp11a1基因表达(P<0.01),8 g·L-1人参-黄芪和1-8 g·L-1熟地-山茱萸都抑制Cyp21a1基因表达(P<0.05),1-8 g·L-1人参-黄芪和0.25-8 g·L-1熟地-山茱萸均抑制了Cyp11b1基因表达(P<0.05).与佛司可林(Forskolin,FSK)组比较,人参-黄芪与熟地-山茱萸预处理Y1细胞后,FSK不能诱导Star、Cyp11a1、Cyp21a1、Cyp11b11基因表达(P<0.01).结论 人参-黄芪促进皮质酮分泌,熟地-山茱萸抑制皮质酮分泌,主要通过调节类固醇激素合成酶表达发挥药效.

目的:研究顺铂短期给药诱发小鼠药源性证候的属性及对类固醇激素合成功能的影响.方法:40只雄性ICR小鼠随机分为对照组及顺铂不同剂量(3、5、10 mg·kg-1·d-1)组,每组10只.各药物干预组小鼠腹腔注射相应剂量的顺铂,连续5 d.分别于干预5 d后和停药5 d后,观察小鼠体征(体质量、摄食量、腋温、爪色泽),并进行行为学测试(抓力、旷场试验和转棒试验).观察期后,取血清;分离肾脏、心脏、胸腺和脾脏,计算脏器指数;收集肾上腺和睾丸.ELISA检测血清皮质酮和睾酮含量;HE染色后光镜下观察肾上腺与睾丸组织形态学变化;PCR、Western blot分别检测肾上腺皮质类固醇激素合成酶和睾丸睾酮合成酶相关的mRNA、蛋白表达.结果:①干预5 d后,顺铂10 mg·kg-1·d-1组小鼠体质量、摄食量、腋温、水平运动及垂直运动较对照组均显著降低(P＜0.05,P＜0.01);顺铂3、5mg·kg-1·d-1组小鼠腋温明显低于对照组,而爪色r值高于对照组(P＜0.05,P＜0.01).停药5 d后,顺铂10 mg·kg-1·d-1组小鼠体质量、摄食量、在棒时间、抓力、腋温、爪色r值、水平运动及垂直运动较对照组均显著降低(P＜0.05,P＜0.01);顺铂3、5 mg·kg-1·d-1组小鼠垂直运动较对照组亦明显减少(P＜0.01).②与对照组相比,10 mg·kg-1·d-1顺铂作用显著抑制小鼠脾脏、胸腺、心脏,脏器指数明显降低(P＜0.05,P＜0.01),而肾脏指数显著升高(P＜0.01);5 mg·kg-1·d-1顺铂作用亦抑制心脏(P＜0.05).③形态学观察显示,顺铂10mg·kg-1·d-1组小鼠肾上腺皮质球状带细胞肿胀明显,顺铂5、10mg·kg-1·d-1组小鼠可见各级生精细胞排列略紊乱,睾丸间质细胞收缩明显.④与对照组相比,顺铂10 mg·kg-1·d-1组小鼠血清皮质酮含量显著升高(P＜0.01),血清睾酮含量无明显变化.⑤与对照组相比,顺铂3 mg·kg-1·d-1组小鼠肾上腺Star、Cyp21a1的mRNA表达受到抑制(P＜0.05),而顺铂10mg·kg-1·d-1组小鼠肾上腺Star、Cyp11b1、Cyp21a1的mRNA表达及StAR、CYP21A2、CYP11B1蛋白表达显著上调(P＜0.05,P＜0.01).⑥与对照组相比,10 mg·kg-1·d-1顺铂作用显著抑制睾丸 StAR、CYP1 1A1、CYP17A1、HSD3B2的 mRNA 与蛋白表达(P＜0.05,P＜0.01);5 mg·kg-1·d-1 顺铂干预显著抑制 Star 和Hsd3b2的 mRNA 表达(P＜0.05,P＜0.01);3 mg·kg-1·d-1 顺铂作用亦显著抑制 Cyp17a1 和Hsd3b2的mRNA表达(P＜0.01).结论:顺铂短期给药可诱发小鼠虚证表现,以精气亏虚为主,其物质基础与类固醇激素合成及储备下降有关.

睾酮的合成与男子生殖功能及衰老相关,睾丸间质细胞是合成与分泌睾酮的主要场所,多种酶与转录因子参与睾酮合成过程.温补肾阳中药及其主要活性成分,包括淫羊藿、菟丝子和肉苁蓉等被证明具有调节生殖内分泌水平,改善氧化应激、抑制炎症发生、调节睾酮合成相关酶mRNA及蛋白表达,影响睾酮合成与分泌.结合文献就影响睾丸间质细胞睾酮合成与分泌的主要因素,及温补肾阳中药对睾酮合成与分泌的调节研究进行综述.

目的 研究黄柏酮对肾上腺皮质激素合成与分泌的影响.方法 体外培养Y1小鼠肾上腺皮质瘤细胞,采用2.5～160μmol/L黄柏酮干预细胞24h,运用MTT法检测细胞活性;以80μmol/L黄柏酮处理细胞24h后采用流式细胞术检测细胞周期,共聚焦显微镜检测细胞线粒体膜变化;以ELISA法检测细胞分泌液皮质酮含量;以40～160μmol/L黄柏酮干预细胞24h,80μmol/L黄柏酮干预细胞24～48h,运用qPCR法检测皮质酮合成酶mRNA表达,Western blot检测其蛋白表达.结果 与对照组比较,2.5～40μmol/L黄柏酮对细胞的抑制率约为35％(P<0.05),160μmol/L黄柏酮对细胞的抑制率为60％以上(P<0.01).黄柏酮导致G1期细胞显著增加并促进线粒体膜亮度增强(P<0.05),同时显著抑制线粒体膜Mfn1、Mfn2蛋白表达以及皮质酮分泌(P<0.05).进而发现,40～160μmol/L黄柏酮显著抑制Cyp11a1、Cyp11b1、Hsd3b2、SF-1基因表达(P<0.01),160μmol/L黄柏酮显著增强Star、Cyp21a1、Nr4a1、Nr4a2基因表达(P<0.01);80μmol/L黄柏酮持续给药48h内,均显著抑制Cyp11a1、Cyp21a1、Cyp11b1、Hsd3b2基因表达(P<0.01),并抑制StAR、CYP11A1蛋白表达,然而促进Star基因表达(P<0.01).结论 黄柏酮抑制肾上腺皮质细胞皮质酮合成过程,可能与其阻滞细胞周期及影响线粒体膜上类固醇激素合成酶表达有关.

目的 研究顺铂不同给药方案对小鼠类固醇激素合成及储备功能的影响.方法 40只雌性ICR小鼠分为4组,顺铂连续和间隔造模组以及两个对照组.顺铂连续造模组给药7 d,每天注射3 mg/kg顺铂;间隔造模组每周注射1次10 mg/kg顺铂,共4次.末次造模前进行旷场实验、转棒疲劳实验、抓力测试;次日处死前称体重,摘取小鼠心脏、脾、胸腺和双侧肾并称重;RT-qPCR检测下丘脑GnRH和CRH、垂体Pomc、Fshb、Lhb、Pou1f1基因表达,以及肾上腺和卵巢类固醇合成酶(StAR、Cyp11a1、Cyp21a1、Cyp17a1、Cyp11b1、Hsd3b2、Cyp19a1、Hsd17b1)及受体(Esr1、Esr2和Gper1)基因表达;Western Blot检测肾上腺和卵巢StAR、CYP11A1、CYP21A2和CYP11B1蛋白表达.结果 与对照组比较:(1)不同频次顺铂造模均可抑制小鼠体重增长,顺铂累积后可以诱导小鼠气虚证表现,并抑制脏器质量.(2)不同频次顺铂造模均能加速激活原始卵泡分化,产生大量闭锁卵泡,诱导膜细胞及颗粒细胞凋亡,健康的成熟卵泡减少;但对肾上腺组织和细胞形态影响不大.(3)连续造模后肾上腺StAR蛋白表达下调;间隔造模后StAR升高,但下调CYP11B1蛋白表达.(4)连续造模后,卵巢Cyp11a1、Hsd3b2、Esr1和Gper1基因表达上调,Star和Cyp17a1基因表达下调;间隔造模后StAR蛋白表达和Hsd3b2基因表达下调.(5)顺铂连续造模后下丘脑GnRH和CRH基因表达明显下调.(6)顺铂连续造模后垂体Pomc、Fshb、Pou1f1基因表达升高,而间隔造模组Lhb表达下降.结论 顺铂能够诱发小鼠气虚、精气不足证的相关表现;合成类固醇激素的卵巢受到选择性损伤大于肾上腺,短期连续给药也进一步抑制HPA轴和HPG轴的高级中枢下丘脑和垂体功能.

通过在实验中医学课程考核方式中引入探究性学习实践,发现学生在理论向科研实践过渡的过程中,暴露出的一系列问题.由此,在学生探究性学习基础上,加入个性化辅导的教学环节,针对每名学生的探究性学习报告,评价其科研设计的优势及薄弱环节、分析问题原因、找到解决思路,帮助学生构建完整的科研思维框架.结果 显示,个性化辅导有助于学生对本课程知识点的巩固,学生探究性学习报告的成绩及科研设计的各方面能力均有显著提升.据此形成的"理论学习—探究性学习(暴露问题)—个性化辅导(解决问题)"教学模式,更有助于学生科研思维的形成,从而实现本课程的教学目标.

目的:对糖皮质激素诱发的小鼠药源性证候模型建立规范化的造模方法及评价标准.方法:对模型动物、糖皮质激素类药物及其干预的剂量、时间等进行考察,采用小鼠辨证论治实验方法学以及反映肾上腺皮质功能的实验指标综合评价小鼠证候属性.结果:对糖皮质激素诱发的小鼠药源性证候模型,建议首选ICR小鼠,常规造模采用3.3 g·kg-1·d-1氢化可的松连续灌胃14 d;大剂量短期造模采用25 g ?kg-1·d-1氢化可的松连续灌胃5 d;小剂量长期造模采用0.33 g·kg-1·d-1氢化可的松连续灌胃28 d;小剂量减停干预造模采用0.66 g·kg-1·d-1氢化可的松连续灌胃给药28 d、再剂量减半7 d、停药14 d.较理想的评价指标包括:体质量、脾脏指数、胸腺指数、腋温、体表红外温度、自主行为活跃度、四肢抓力、血液皮质酮、肾上腺类固醇激素合成酶特异性分子(StAR、CYP11A1、CYP11B1、CYP21A1、SRBI、LDLR)、肾上腺皮质形态及超微结构.结论:对糖皮质激素诱发的小鼠药源性证候模型建立了规范化的造模方法及可量化的疗效评价标准,为中药及其复方的药效研究提供理想的小鼠证候模型.

原发性肝癌(PLC)是我国难治性恶性肿瘤,病死率高[1].PLC的治疗,目前仍以早期局部手术切除作为首选方案.但是,所有促进早期手术得以实现的努力和各种疗法的实施,使得术后1、3、5年生存率已接近高限[2].此外,国际上多采纳巴塞罗那临床肝癌分期标准,对B期(单发或多发的大肿瘤)推荐经导管肝动脉化疗栓塞(TACE),对C期(伴大血管/胆管癌栓)仅推荐多靶向性药物索拉非尼[3,4].有研究表明,我国约80％的肝癌患者都不同程度地接受过中医药治疗[5,6].而中医药在预防和降低肝癌复发转移率、改善症状、提高生存质量,甚至延长患者生存期等方面具有独特优势[5,6].