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.

目的 探索α-常春藤皂苷协同雷公藤红素对SMMC7721肝癌细胞增殖和周期的影响.方法 MTT法检测细胞存活率,CompuSyn软件获取相互作用指数(CI)并筛选出最佳协同组合.流式细胞仪检测细胞凋亡和周期分布,Western blot法检测细胞凋亡相关蛋白Bax和Bcl-2、周期蛋白依赖性激酶(CDKs)和细胞周期蛋白(Cyclins)的表达.结果 单用α-常春藤皂苷或雷公藤红素均能抑制细胞增殖,且呈剂量依赖性;作用48 h后,当α-常春藤皂苷质量浓度为10.0μg/mL,雷公藤红素质量浓度为0.30 μg/mL时,CI值为0.24,协同效应最佳.在以上质量浓度条件下,协同组早期凋亡率高于其余3组(P＜0.05).α-常春藤皂苷组及协同组晚期凋亡率高于对照组(P＜0.05);总凋亡率以协同组最高且高于对照组(P＜0.05).雷公藤红素及协同组G0/G,期比例较对照组升高(P＜0.05);α-常春藤皂苷组S期比例较对照组升高(P＜0.05);协同组G1+S期总比例最高且高于对照组(P＜0.05).与对照组比较,各给药组Bax、Bcl-2及Bax/Bcl-2比值无明显变化(P＞0.05);协同组CDK2和CDK4蛋白表达降低(P＜0.05);雷公藤红素组和协同组Cyclin D1表达均升高(P＜0.05);各给药组Cyclin E1表达均升高(P＜0.05).结论 α-常春藤皂苷协同雷公藤红素抑制SMMC7721细胞增殖效应确切,该效应可能与诱导G0/G,期周期阻滞、G1+S期总比例增加有关,而Cyclin D1可能是参与其中的关键分子.

目的 观察预知子联合雷公藤红素对肝癌荷瘤裸鼠的抑瘤作用及对核因子(NF)-κB通路的影响,并探讨其作用靶点.方法 皮下接种肝癌SMMC7721细胞建立荷瘤裸鼠模型,将裸鼠随机分为模型对照组、预知子提取物组、雷公藤红素组和联合组,将药物包载于壳聚糖/甘油磷酸钠温敏性凝胶系统,分别于分组后第1、6、11日经皮瘤内注射相应药物,共3次.每2日测量裸鼠体质量和瘤径,第16日称量瘤体及肝、脾、左肾脏器质量,计算瘤指数、抑瘤率及脏器指数;ELISA测定血清肿瘤坏死因子(TNF)-α含量;HE染色观察瘤组织形态变化;Western blot检测瘤组织NF-κB通路相关蛋白表达.结果 与模型对照组比较,各给药组裸鼠体质量、去瘤体质量、瘤体积、瘤质量和瘤指数均呈下降趋势,其中以雷公藤红素组最为显著(P＜0.05),联合组瘤体积和瘤质量显著下降(P＜0.05),各给药组裸鼠肝和左肾质量及指数均显著下降(P＜0.05),预知子提取物组、雷公藤红素组和联合组抑瘤率分别为6.68％、35.08％和22.00％;各给药组裸鼠血清TNF-α含量显著增加(P＜0.05);HE染色显示,各给药组裸鼠瘤组织均出现间质增多现象,且联合组部分区域出现炎症细胞浸润、堆积;Western blot结果显示,雷公藤红素组裸鼠瘤组织IκB激酶α(IKKα)、周期蛋白依赖性激酶2(CDK2)、周期蛋白D1(CyclinD1)、NF-κBp65蛋白表达显著降低(P＜0.05),联合组裸鼠瘤组织NF-κB抑制蛋白α(IκBα)和p-IκBα蛋白表达显著降低(P＜0.05).结论 预知子联合雷公藤红素能抑制肝癌荷瘤裸鼠肿瘤生长,NF-κB通路p-IκBα分子可能是二者联合发挥抑瘤作用的重要靶点.

肝癌属难治性恶性肿瘤,癌细胞增殖迅速且拥有抵御药物干预的复杂分子应激网络,是其难以治愈的重要原因.发展包含中医药在内的综合治疗手段,开发新药(如中药提取物或有效成分),开展联合用药研究,具有一定的应用前景.通过综述中药成分联用抑制肝癌细胞增殖及机制研究,以期为肝癌的综合治疗及中医药相关研究提供参考.

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.

目的 观察预知子醇提物联合雷公藤红素对SMMC7721细胞增殖和凋亡的影响,探讨其可能的作用机制.方法 将不同浓度预知子醇提物和雷公藤红素分别作用于SMMC7721细胞72 h,MTT法获得细胞存活率、72 h IC50后,计算药物相互作用指数(CDI).将SMMC7721细胞分为空白对照组、预知子醇提物组、雷公藤红素组和预知子醇提物+雷公藤红素组(联合组),光镜观察细胞形态,荧光显微镜观察细胞核形态,流式细胞术检测细胞凋亡率,Western blot检测线粒体凋亡途径、内质网应激途径、自噬相关和线粒体-内质网结构偶联(MAM)相关蛋白的表达.结果 预知子醇提物和雷公藤红素72 h IC50剂量CDI值为0.45,存在显著协同效应.联合组细胞存活率明显降低(P<0.05),细胞形态发生改变,且部分细胞核呈碎块状,细胞凋亡率较其余3组显著升高(P<0.05).与空白对照组比较,雷公藤红素组和联合组线粒体凋亡相关蛋白Caspase-3表达明显降低(P<0.05),联合组cleaved Caspase-3表达明显降低(P<0.05),预知子醇提物组内质网应激相关蛋白Grp78表达明显降低(P<0.05),雷公藤红素组和联合组Grp78蛋白表达明显升高(P<0.05);联合组Chop蛋白表达较其余3组均明显升高(P<0.05);与空白对照组比较,预知子醇提物组和联合组自噬相关蛋白p62、LC3BⅡ及LC3BⅡ/Ⅰ比值均显著升高(P<0.05),各给药组MAM标志物Mfn2表达均升高但差异无统计学意义(P>0.05).结论 预知子醇提物联合雷公藤红素可抑制SMMC7721细胞增殖、诱导细胞凋亡,预知子醇提物和雷公藤红素作用靶点不尽相同;二者的一致性效应机制方面,除Caspase-3和cleaved Caspase-3、Chop和Mfn2等分子外,尚需进一步扩大关注面.

目的 探讨预知子醇提物(AFE)与姜黄素(CUR)联用抑制SMMC7721肝癌细胞增殖协同效应及机制.方法 将不同浓度预知子醇提物AFE(0、0.4、0.8、1.2、1.6、2.0 g·L-1)和姜黄素CUR(25.0、20.0、15.0、10.0、5.0μmol·L-1)分别作用于SMMC7721肝癌细胞72h,噻唑蓝法(MTT)获得细胞存活率;在此基础上进一步设置不同浓度9种组合,以药物相互作用指数(Coefficient of Drug Interaction,CDI)为标准筛选最佳协同效应组合,同时设置空白以及各自等剂量单药对照组,进一步采用光镜观察细胞形态、流式细胞术(FCM)检测细胞凋亡,蛋白免疫印迹法(WB)检测线粒体和内质网应激凋亡、死亡受体途径相关蛋白表达量.结果 ①AFE 72h IC50为2.09 g·L-1;CUR 72h IC50为16.90μmol·L-1;②AFE 1.5 g·L-1+CUR 15.0μmol·L-1组合协同效应最佳,CDI值为0.74±0.06;③细胞形态方面,AFE作用后细胞增殖被抑制且出现ERS空泡样表现,CUR作用后细胞增殖被抑制但无相同形态变化,协同组类似二者效果的叠加;④AFE、CUR单独作用凋亡现象基本不出现,协同组诱导细胞凋亡现象则被显著增强,凋亡率显著高于其它3组(P<0.05);⑤协同组Bcl-2较对照和CUR组显著上升(P<0.05);AFE下调Caspase-9表达(P<0.05);Cleaved Caspase-9和Parp1均呈下降趋势,其中大片段Cleaved Parp1和小片段Cleaved Parp1均被CUR显著下调(P<0.05),但后者被AFE和协同组显著上调(P<0.05);AFE显著下调Bip表达(P<0.05);Chop在各用药组中表达量均被下调(P<0.05),且在协同组低于各单药组(P<0.05);p-JNK在AFE和协同组均被上调(P<0.05);IKBα在CUR和协同组均被上调(P<0.05),且协同组高于各单药组(P<0.05).结论 预知子醇提物和姜黄素联合抑制肝癌细胞增殖协同效应确切;线粒体和内质网应激凋亡、死亡受体途径上,Bcl-2,Parp1,Chop、IKBα等诸多标志性分子可能参与该过程.

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.

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].

Objective:To evaluate the teaching effect of moral education materials implied in scientific research cases in the teaching of "Experimental Traditional Chinese Medicine (TCM)".Methods:The moral education materials implied in scientific research cases of "Experimental TCM" were extracted and skillfully integrated into the teaching of professional knowledge. A questionnaire survey was conducted with questions as "whether it is good to demonstrate the potential humanistic spirit by case teaching, whether this helps improve your interest in science and whether this teaching form affects your study, life and work attitude in the future" "Ten specific items from scientists' moving deeds that touch students and their recognitions" to assess the teaching effect.Results:95.8 percent of students affirmed this teaching form and thought it helped improve their interest in scientific exploration. 87.5 percent of students considered the humanistic spirit would affect their study, life and work in the future. 77.1-89.6 percent of students held a positive attitude to the ten items derived from the scientists' moving stories. Among these items, the percentages of the two items, "the spirit of being able to endure loneliness, work hard to make contributions selflessly without seeking for rewards or reputations" and "having respect forpeople and their scientific research achievements with courage to challenge the authorities of scientific research" are the lowest and highest, respectively.Conclusion:It is good to apply the moral education materials implied in the scientific research cases in the teaching of "Experimental TCM", which basically achieves the teaching goal, but there is still room for improvement.

Experimental Traditional Chinese Medicine is an emerging discipline that plays an important role in cultivating innovative talents of traditional Chinese medicine (TCM). In recent years, with the rapid development of TCM and the new requirements of positioning, and also combined with the college students' cognitive characteristics, we have revised and republished the "Experimental TCM" (Third Edition) textbook, which focuses on introducing knowledge by adopting relevant scientific research cases. This test book was used in the teaching of undergraduates of batch 2013 in the eight-year program in Shanghai University of Traditional Chinese Medicine. After-class questionnaires showed that this teaching mode, guided by scientific research case, is not only helpful for students to develop their quality and ability of adopting modern experimental methods initially in the study and development of TCM, but also able to spread the great achievements of TCM researches. The teaching mode is also conducive to enhancing students' sense of responsibility for the modernization of TCM. Therefore, it is suggested that the course of Experimental TCM should be promoted in the colleges and universities of TCM.

ETHNOPHARMACOLOGICAL RELEVANCE:Hepatocellular carcinoma (HCC) is an aggressive malignancy with increasing mortality in China. Screening and identifying effective anticancer compounds from active traditional Chinese herbs for HCC are in demand. Akebia trifoliata (Thunb) Koidz, with pharmacological anti-HCC activities in clinical, has been shown in previous research. In the present research, we elucidated a potential anticancer effect of Akebia saponin E (ASE), which is isolated from the immature seeds of Akebia trifoliata (Thunb.) Koidz, and revealed that ASE could induce severe expanded vacuoles in HCC cells. But the potential mechanism of vacuole-formation and the anti-HCC effects by ASE remain uncover. AIM OF THIS STUDY:To elucidate the potential mechanism of vacuole-formation and the proliferation inhibition effects by ASE in HCC cell lines. MATERIALS AND METHODS:MTT assay, colony formation assay and flow cytometry were performed to detect cell viability. Immunofluorescence analysis was used to examine the biomarkers of endomembrane. Cells were infected with tandem mRFP-GFP-LC3 lentivirus to assess autophagy flux. RNA-seq was conducted to analyze the genome-wide transcriptional between treatment cell groups. In vitro PIKfyve kinase assay is detected by the ADP-GloTM Kinase Assay Kit. RESULTS:ASE could inhibit the proliferation of HCC with severe expanded vacuoles in vitro, and could significantly reduce the size and weight of xenograft tumor in vivo. Further, the vacuoles induced by ASE were aberrant enlarged lysosomes instead of autophagosome or autolysosomes. With cytoplasmic vacuolation, ASE induced a mTOR-independent TFEB activation for lysosomal biogenesis and a decrement of cholesterol levels in HCC cells. Furthermore, ASE could reduce the activity of PIKfyve (phosphoinositide kinase containing a FYVE-type finger), causing aberrant lysosomal biogenesis and cholesterol dyshomeostasis which triggered the expanded vacuole formation. CONCLUSION:ASE can prospectively inhibit the kinase activity of PIKfyve to induce lysosome-associated cytoplasmic vacuolation, and may be utilized as an alternative candidate to treat human HCC.

为提升实验中医学学习成果的探究性和个性化,课程组对课程的考核模式进行了改革,在原来单一的闭卷考核中引入探究性学习的考核方式(占总分的25％).由教师根据课程的教学内容和培养目标要求学生提出一个假说并设计实验进行验证,学生根据自己兴趣,开展探究性的学习,并形成学习报告用于成绩评定.分析发现,探究性学习的成绩与闭卷考核成绩具有显著的正相关性;在探究性学习中,学生重点关注的是失眠、抑郁等精神神经相关疾病,更倾向于开展中药复方作用于相关疾病动物的药效药理实验.通过探究性学习,学生的探究能力、知识点融会贯通等能力得到了提高,培养并开拓了研究的思维,同时也暴露出一些问题,有待后续进一步完善和优化.

在"大思政"的格局下,为了更深入地挖掘本门课程的育人职能,从德育素材的收集、融合方式的设计、教师育人意识的提高等方面探索课程德育的教学方式,并从教师和学生两方面进行评价和分析.结果表明实验中医学课程德育对学生正确人生观的树立产生积极的影响,对提高中医专业学生思想道德水平和专业自信发挥了正向促进作用,提高了学生的学习效率.同时,实施课程德育使专业课教师的思政育人能力得到锻炼和提升.

Ethnopharmacological relevance: The high recurrence rate postoperative and extensive metastases have become the obstacle of Hepatocellular Carcinoma (HCC) efficacy improvements, which contribute to most of the patient mortality. Akebia trifoliata (Thunb.) Koidz has been shown pharmacological activities in clinical and anti-HCC biological activity in previous research, but its potential function of anti-metastasis remains unknown. Aim of this study: To make sure whether ATKSE inhibits migration and invasion in HCC cell lines in vitro and the potential mechanism. Materials and methods: A UHPLC-HRMS analysis was adopted to identify and control the quality of the ethanol extract of Akebia trifoliata (Thunb.) Koidz Seed (abbreviated ATKSE). Cell viability of three kinds of HCC cell lines (HEPG2, HUH7, and SMMC7721) was detected using MTT assay and Flow cytometry. Adhesion capacity was measured by cell-matrigel adhesion assay. Wounded healing and Matrigel-transwell invasion assays were performed to assess cell migration and invasion, respectively. Western blot assay was used to detect several metastasis-related protein molecules, including FAK adhesion signaling, cadherin molecules, and MMPs. ELISA assay was used to evaluate the secreted MMP9 level. Results: ATKSE significantly suppressed HCC cells viability and proliferation (from 0.9 up to 3.0 mg/ml); then under sub-lethal concentration (from 0.25 up to 1.0 mg/ml), ATKSE inhibited cell adhesion, migration, and invasion in a way of dose-dependent. Several metastatic-related molecules or pathway, including FAK adhesion signaling, cadherin molecules, and MMPs, took part in this process. There are both differences and commonalities in various cell lines: typically such as p-FAK was down-regulated by ATKSE in both HEPG2 and SMMC7721, while was raised in HUH7; Further attempts on the combination of ATKSE and FAK inhibitors, provide us with the enhanced inhibitory effects of invasion and migration in HEPG2 and HUH7 cells, as well as antagonistic effects in SMMC7721. As a target or potential mechanism, it may be more valuable to concern FAK inhibition by ATKSE in HEPG2 cells than in the other two cells. Conclusions: These results suggest that ATKSE has anti-metastasis potency in HCC cells.

Psoralen is a coumarin-like and coumarin-related benzofuran glycoside, which is a commonly used traditional Chinese medicine to treat patients with kidney and spleen-yang deficiency symptom. Psoralen has been reported to show estrogen-like activity, antioxidant activity, osteoblastic proliferation accelerating activity, antitumor effects and antibacterial activity. However, the antitumor mechanism of psoralen is not fully understood. This study aimed to investigate the therapeutic efficacy of psoralen in human hepatoma cell line SMMC7721 and the mechanism of antitumor effects. Psoralen inhibited proliferation of SMMC7721 in a dose- and time-dependent manner, and promoted apoptosis. Further, psoralen activated the ER stress signal pathway, including the expansion of endoplasmic reticulum, increasing the mRNA levels of GRP78, DDIT3, ATF4, XBP1, GADD34 and the protein levels of GDF15, GRP78, IRE1α, XBP-1s in a time-dependent manner. Psoralen induces cell cycle arrest at G1 phase by enhancing CyclinD1 and reducing CyclinE1 expression. Moreover, TUDC couldn’t inhibit the psoralen-induced ER stress in SMMC7721 cells. Psoralen can inhibit the proliferation of SMMC7721 cells and induce ER stress response to induce cell apoptosis, suggesting that psoralen may represent a novel therapeutic option for the prevention and treatment hepatocellular carcinoma.

目的:观察预知子种子提取物抑制HepG2肝癌细胞的增殖及黏附作用,探讨其相关机制.方法:将中药预知子种子醇提物(1.5,1.8,2.1,2.4,2.7,3.0 g·L-1)和整合素抑制剂西仑吉肽(Cilengitide)(0.5,1.0,2.0,4.0,8.0,32 μmol·L-1)作用于HepG2肝癌细胞,噻唑蓝(MTT)比色法获得各药物作用24 h后的细胞存活率检测结果,并依据该结果设置组别和用药`浓度,中药组(预知子醇提物1.8 g·L-1),Cilengitide组(Cilengitide,1.5 μmol·L-1),半量联合用药组(0.9 g·L-1+0.75 μmol·L-1),作用24 h后观察对细胞-基质黏附能力的影响,以及相关黏附分子蛋白相对表达量的变化.结果:MTT实验结果显示,不同药物处理HepG2细胞24 h后,中药1.8 g·L-1组细胞黏附率在86.8％左右,Cilengitide1.5 μmol· L-1组细胞存活率在89.4％左右.细胞-基质黏附实验中,空白组细胞-基质黏附率为(100.0±11.5)％,中药组为(32.3±14.0)％,Cilengitide组为(33.8±5.7)％,半量联合用药组为(33.4±6.4)％,各用药组细胞存活率显著低于空白组(P＜0.01).与空白组比较,中药组小亚基钙蛋白酶1(CAPNS1),硒蛋白P(SEPP1)和整合素α2(ITGA2)蛋白的表达量均上升(P＜0.05);Cilengitide组SEPP1蛋白表达量上升(P＜0.05),整合素αV(ITGAV)蛋白的表达量下降(P＜0.01);半量联合组CAPNS1,SEPP1蛋白表达量均上升(P＜0.05),ITGAV蛋白表达量下降(P＜0.05).结论:预知子种子醇提物能抑制HepG2肝癌细胞的增殖及黏附作用;和Cilengitide联合用药后存在减量增效潜能;以上黏附分子蛋白表达量的变化,可能是预知子的部分作用机制.

目的 探讨微信辅助《中医学综合实验》基于问题学习(problem-based learning,PBL)教学模式的教学效果.方法 在2011级中医七年制学生的《中医学综合实验》PBL教学中引入微信作为一种辅助教学工具.课程结束时发放并回收调查问卷,统计问卷调查中的客观选项,并分析学生的主观感想.结果 (1)所有学生都认可微信辅助PBL教学的模式,课内和课外互动非常活跃.(2)除有一位学生对“是否有助于激发学生的好奇心、调动主观能动性”持否定态度外,其他所有学生认为该教学模式对其他各项能力的培养均有帮助.(3)与以往该课程单独PBL教学比较,微信辅助教学后课程的教学内容和互动在课外均得到了有效的延伸,反过来进一步提高了PBL课堂的教学效果.结论 微信辅助《中医学综合实验》PBL教学的教学模式达到了预期的教学目标,但是否将学生在微信上的表现作为该课程的评价内容之一,还需进一步探索.

Objective To observe the effects of Akebia Fruit seed (AFS) extract on morphology of different hepatocellular carcinoma cells.Methods By the comprehensive application of cell biology,scanning electron microscopy(SEM) and transmission electron microscopy(TEM),the effects of alcohol extract of AFS on the morphological characteristics of HepG2,Huh 7,and SMMC7721 were observed.Results Untreated HepG2 cells show overlapping growth and the surfaces are fully covered with villus.After the treatment with AFS extract,the partial cell villus atrophied and disappeared,the cells became irregular and even collapsed,and rough endoplasmic reticulum(ER) expanded to different degrees.Spontaneous apoptosis could form in Huh 7 cells.After the treatment with AFS extract,the irregular villus,cell shrinking,a lot of apoptotic bodies,and numerous vacuoles caused by endoplasmic reticulum stress were observed in cells.Untreated SMMC-7721 cells are fully covered with longer villus.After the treatment with AFS extract,abnormal morphology could be detected.The cell body expansion with irregular round shape were observed,some regions on the surface of cells collapsed or upheaved,the surface of some cells collapsed due to severe endoplasmic reticulum stress,and a large number of heavily expanded endoplasmic reticulum were commonly found.Conclusion These three kinds of hepatocellular carcinoma cell lines exhibit various morphological characteristics,but serious ER stress with different degrees can be induced after the treatment with Akebia Fruit seed extract,which may be one way of Akebia Fruit seed acting on liver cancer.