Additional file 7. Correlation of UPR-related genes and EIF5A2.

Additional file 6. Representative GO biological processes of down-regulated DEGs.

Context Lucialdehyde B (LB), an effective triterpenoid isolated from Ganoderma lucidum (Leyss. ex Fr.) Karst. (Polyproraceae), exerts cytotoxic activity against nasopharyngeal carcinoma CNE2 cells. Objective To investigate the antiproliferative and pro-apoptotic effects of LB on CNE2 cells and explore its underlying mechanisms. Materials and methods LB concentrations of 5-40 mu g/mL were used. Cell proliferation was determined using MTT, CFSE, and colony formation assays. LB-induced apoptosis and cell cycle arrest were measured by flow cytometry after 48-h LB treatments. Fluorescence microscopy and flow cytometry were performed to measure the alteration of MMP, mPTP opening, ROS level, and Ca2+ content in CNE2 cells. Western blotting was performed to evaluate the expression of mitochondrial apoptosis-related and Ras/ERK signaling proteins. Results IC50 values of LB against CNE2 cells for 24, 48, and 72 h were 25.42 +/- 0.87, 14.83 +/- 0.93, and 11.60 +/- 0.77 mu g/mL, respectively. The CFSE assay showed that the cell proliferation index was 12.70 in the LB treatment group and 31.44 in the control group. LB significantly reduced clonogenic capacity, promoted cell apoptosis and induced cell cycle arrest at the G2/M phase. Our observations also revealed that LB induced ROS and calcium aggregation, opening of mPTP, MMP reduction, upregulation of mitochondrial apoptosis-related protein expression and inhibition of Ras/ERK signaling cascades. Discussion LB suppresses proliferation and induces mitochondrial-dependent apoptosis in nasopharyngeal carcinoma CNE2 cells. Conclusions LB may have a potential use as a clinical drug candidate for nasopharyngeal carcinoma treatment.

Additional file 5: Table S5. Classification of all RASE events between sample groups.

目的:研究灵芝所含有的化学成分,并探究其对人的鼻咽癌CNE2细胞的细胞毒活性.方法:(1)利用实验室硅胶柱色谱、制备型高效液相色谱法将灵芝进行系统分离纯化,再由波谱解析以及化学方法等相关手段进行结构鉴定;(2)经MTT法检验人的鼻咽癌CNE2细胞被灵芝化学成分增殖抑制的作用,并筛选其中活性成分.结果:(1)本实验从灵芝里分离出6个三萜类的化合物和1个甾醇类的化合物,分别是ganodermanontriol、ergosterol peroxide、ganoderol B、ganodermanondiol、ganoderone A、lucialdehyde C 以及 lucialdehyde B.(2)其中浓度为 5～80 μg·mL-1的区间里,人鼻咽癌CNE2细胞生长受化合物1、3的抑制作用十分弱,而其他的化合物都显现出一定的细胞毒活性,并呈浓度依赖性抑制肿瘤细胞生长.人鼻咽癌CNE2细胞受化合物2、4、5、6和7作用48 h的IC50值分别是(42.80±1.16)μg·mL-1、(64.35±1.20)μg·mL-1、(55.30±1.32)μg·mL-1、(15.33±1.21)μg·mL-1、(14.17±1.10)μg·mL-1.结论:由灵芝分离出的三萜成分lucialdehyde C、lucialdehyde B对人鼻咽癌CNE2细胞生长具有较强的抑制作用,有希望成为灵芝治疗鼻咽癌的活性成分.

Two sterols and seven triterpenoids were isolated and identified from Ganoderma lucidum by silica gel column chromatography, preparative high-performance liquid chromatography and spectra analysis. Then, the multidrug resistance reversal activities of these compounds were assessed using MTT assay. Among these compounds, ganoderol B (3), ganoderone A (4), ganodermanondiol (6) and ganoderiol F (8) were shown to reverse the resistance of human oral epidermoid carcinoma cell line KBv200 to doxorubicin, and the reversal folds were 6.59, 4.70, 4.01 and 7.09, respectively. Ganoderiol F could increase the intracellular accumulation of doxorubicin in KBv200 cells through inhibiting P-glycoprotein transport function. Further mechanistic investigation found that ganoderiol F did not alter P-glycoprotein expression. In conclusion, ganoderiol F has potent effect in reversing P-glycoprotein mediated tumor multidrug resistance. Potential reversal agents against multidrug resistance in tumor may be found in triterpenoids from Ganoderma lucidum.[Formula: see text].

目的 从灵芝中制备总黄酮,并探讨其体外细胞毒活性.方法 回流提取法结合聚酰胺柱色谱法从灵芝中制备总黄酮提取物;紫外-可见分光光度法测定醇浸膏及总黄酮提取物中总黄酮含量;MTT法测试灵芝总黄酮提取物的细胞毒活性.结果 灵芝醇提浸膏与聚酰胺按质量比为1:1混匀后装柱,先用40％乙醇洗脱,再用无水乙醇洗脱,收集无水乙醇洗脱液,减压浓缩蒸干得灵芝总黄酮提取物.醇浸膏中含总黄酮含量为8.98％,灵芝总黄酮提取物中总黄酮含量为18.09％.灵芝总黄酮提取物在体外对人胰腺癌MIA-PACA-2和人乳腺癌MDA-MB-231肿瘤细胞的增殖有抑制作用,IC50分别为(123.7±4.1)μg·mL-1和(196.1±2.5)μg·mL-1.对HepG2、SMMC7721和HL-60细胞的增殖抑制作用较弱.结论 聚酰胺可富集灵芝中总黄酮,灵芝总黄酮提取物对人胰腺癌MIA-PACA-2细胞和人乳腺癌MDA-MB-231细胞有一定细胞毒活性.

Summary Background Coronavirus disease 2019 (COVID-19) is an emerging infectious disease.It was first reported in Wuhan, China, and then broke out on a large scale around the world.This study aimed to assess the clinical significance of two different nutritional indices in 245 patients with COVID-19. Methods In this retrospective single-center study, we finally included 245 consecutive patients who confirmed COVID-19 in Wuhan University Zhongnan Hospital from January 1 to February 29. Cases were classified as either discharged or dead. Demographic, clinical and laboratory datas were registered, two different nutritional indices were calculated: (i)the Controlling nutritional status (CONUT) score; (ii) prognostic nutritional index (PNI). We used univariate and multivariate logistic regression analysis to explore the relationship between nutritional indices and hospital death. Results 212 of them were discharged and 33 of them died. In-hospital mortality was signifcantly higher in the severe group of PNI than in the moderate and normal groups. It was also significantly worse in the severe-CONUT group than in the moderate-, mild-, and normal-CONUT groups. Multivariate logistic regression analysis showed the CONUT score (odds ratio3.371,95%CI (1.124–10.106), p = 0.030) and PNI(odds ratio 0.721,95% CI (0.581–0.896), P=0.003) were independent predictors of all-cause death at an early stage; Multivariate logistic regression analysis also showed that the severe group of PNI was the independent risk predictor of in-hospital death(odds ratio 24.225, 95% CI(2.147–273.327), p=0.010).The CONUT score cutoff value was 5.5 (56.00 and 80.81%; AUC 0.753; 95% CI (0.644–0.862); respectively). The PNI cutoff value was 40.58 (81.80 and 66.20%; AUC 0.778; 95% CI (0.686–0.809); respectively). We use PNI and the COUNT score to assess malnutrition, which can have a prognosis effect of COVID-19patients. Conclusion The CONUT score and PNI could be a reliable prognostic marker of all-cause deathin patients with COVID-19.

As a noted medicinal mushroom, Ganoderma lucidum ( G. lucidum ) has been reported to have a number of pharmacological effects such as anti-tumor and liver protection. Compared with the common ethanol reflux method, supercritical CO 2 extraction has obvious advantages in obtaining antitumor extracts from G. lucidum fruiting body such as short extraction time, low temperature and no solvent residue. However, Using high-pressure supercritical CO 2 without entrainer to obtain the antitumor extracts from G. lucidum and studying their anti-hepatoma effect have not been reported. In this study, high-pressure supercritical CO 2 extracts obtained under 65, 85, and 105 MPa pressure named as G65, G85, G105 respectively and ethanol reflux extract (GLE) were used to investigate their anti-hepatoma activity and the underlying molecular mechanism. The total triterpenoid content of G85 was significantly higher than that of G65 and GLE, but did not differ significantly from that of G105 by UV and high-performance liquid chromatography. GLE, G65, and G85 could inhibit cell proliferation, arrest cell cycle in G2/M phase, and induce apoptosis in two liver cancer cell lines (QGY7703 and SK-Hep1), of which G85 had the strongest effect. The results showed that the potency of their cytotoxicity of the high-pressure supercritical CO 2 extracts on human hepatoma carcinoma cells in vitro was consistent with their total triterpenoid content. G85 exhibited significant anti-hepatoma effect with low toxicity In vivo . Further mechanistic investigation revealed that the anti-tumor effect of these extracts was associated with their inhibition of Ras/Raf/MEK/ERK signaling pathway. Our findings suggest that the high-pressure supercritical CO 2 extraction of G. lucidum fruiting body can be used to obtain a triterpenoid-rich anti-tumor agent, which may have potential clinical significance for the treatment of human hepatoma.

A new demethyl abietane diterpenoid, Triptotin K (3) together with three known compounds, friedelin (1), canophyllal (2), and triptonoterpene (4) were isolated from the roots of Tripterygium wilfordii Hook. f. by silica gel column and preparative high performance liquid chromatography. Their structures were determined by extensive NMR data and mass spectroscopic analysis. Triptotin K showed cytotoxic activities against KB, KBv200, HepG2, and MCF-7/ADM cells lines with IC50 values of 29.88, 36.50, 39.55, and 41.38 μM, respectively.

Overexpression of ATP-binding cassette (ABC) transporters, such as ABCB1 and ABCG2, has been proved to be a major trigger for multidrug resistance (MDR) in certain types of cancer. A promising approach to reverse MDR is the combined use of nontoxic and potent ABC transporters inhibitor with conventional anticancer drugs. We previously reported that FW-04-806 (conglobatin) as a novel Hsp90 inhibitor with low toxicity, capable of attenuating Hsp90/Cdc37 /clients interactions and producing antitumor action in vitro and in vivo. Our early activity screening found that FW-04-806 at non-cytotoxic concentration was able to enhance the cytotoxicity of chemotherapeutic agents on the ABCB1 overexpressing cells. Therefore, we speculated that FW-04-806 might be a promising MDR reversal agent. In the present study we further investigated its reversal effect of MDR induced by ABC transporters in vitro and in vivo. MTT assay in vitro and xenograftes in vivo were used to investigate reversal effect of FW-04-806 on MDR in ABCB1 or ABCG2 overexpressing cancer cells. To understand the mechanisms for the MDR reversal, we examined the effects of FW-04-806 on intracellular accumulation of doxorubicin (DOX, adriamycin, adr)/Rhodamine 123 (Rho 123), efflux of doxorubicin, expression levels of gene and protein of ABCB1 or ABCG2 and ATPase activity of ABCB1, and carried out molecular docking between FW-04-806 and human ABCB1. The results indicated that FW-04-806 significantly enhanced the cytotoxicity of substrate chemotherapeutic agents on the ABCB1 or ABCG2 overexpressing cells in vitro and in vivo suggesting its reversal MDR effects. FW-04-806 increased the intracellular accumulation of DOX or Rho123 by inhibiting the efflux function of ABC transporters in MDR cells rather than in their parental sensitive cells. However, unlike other ABC transporter inhibitors, FW-04-806 had no effect on the ATPase activity nor on the expression of ABCB1 or ABCG2 on either mRNA or protein level. Molecular docking suggested that FW-04-806 may have lower affinity to the ATPase site, which was consistent with its no significant effect on the ATPase activity of ABCB1; However FW-04-806 may bind to substrate binding site in TMDs more stably than substrate anticancer drugs therefore obstruct the anticancer drugs pumped out of the cell. FW-04-806 is a compound that has both anti-tumor and reversal MDR effects, and its antitumor clinical application is worth further study.

天然药物化学是药学专业一门重要的必修课程.其实验课以培养学生的综合实验技能及提高学生的创新能力为主.针对天然药物化学实验教学中存在的教学方法单一的问题,从以问题为中心教学、应用比较实验、引入思维实验和增加互动讨论4个方面对其教学方法进行探讨与实践,探索多样化教学方式在天然药物化学实验教学中的应用效果.实施多样化方法的教学组相对于单一模式教学组学生实验技能掌握与提高较好;问卷调查结果显示大部分学生对于多样化教学方式比较满意.说明天然药物化学实验多样化教学激发了学生的学习兴趣,培养了学生的实验操作技能、科研素养以及创新精神,提高了天然药物化学实验教学效果和质量.

BACKGROUND:Casitas B-lineage lymphoma proto-oncogene-b (CBLB) influences the threshold of T cell activation and controlling peripheral T cell tolerance. In the present study, we hypothesize that potentially functional single nucleotide polymorphisms (SNPs) in CBLB are associated with clinical outcomes in patients advanced non-small cell lung cancer (NSCLC) treated with the first-line chemotherapy.METHODS:We genotyped three SNPs (rs2305035, rs3772534 and rs9657904) at CBLB in 116 advanced NSCLC patients with progression free survival (PFS) data and 133 advanced NSCLC patients with overall survival (OS) data, and we assessed their associations, 95% confidence interval (CI), with clinical outcomes by using Cox proportional hazards regression analyses. In silico functional analysis was also performed for the SNPs under investigation.RESULTS:We found that associations between the three SNPs and PFS/OS were not significant in the overall NSCLC patients. The rs2305035 AA genotype was associated with a worse PFS in female patients and those of non-smokers or light smokers (95% CI, 1.14-11.81, P=0.030; 95% CI, 1.42-10.24, P=0.008; and 95% CI, 1.39-9.93, P=0.009; respectively), compared with the GG+AA genotypes. We also found that the rs9657904 CC genotype was significantly associated with a worse OS than TT + TC genotypes in male advanced NSCLC patients. Further in silico functional analysis revealed that the rs965704 T allele was significantly associated with lower mRNA expression levels of the CBLB gene.CONCLUSIONS:Our findings identified two CBLB SNPs (rs2305035 and rs9657904) that were significantly associated with PFS and OS in several subgroups of Chinese advanced NSCLC patients after the first-line chemotherapy.

BACKGROUND Non-small cell lung carcinoma (NSCLC) mainly includes lung squamous cell carcinoma and adenocarcinoma. This study aimed to investigate the difference between the expression of Cbl-b in lung squamous cell carcinoma and adenocarcinoma. MATERIAL AND METHODS The clinical features and survival data of NSCLC patients and Cbl-b mRNA (FPKM) were obtained from the TCGA database. Then, lung squamous cell carcinoma and adenocarcinoma cell lines were transfected with lentivirus-mediated RNA interference vector to knockdown the expression of Cbl-b. Next, a Transwell assay was performed to study the effect of Cbl-b shRNA on migration and invasion of lung squamous cell carcinoma and adenocarcinoma cells. Finally, Western blot analysis was performed to measure the expressions of PI3K, p-PI3K, AKT, p-AKT, ERK1/2, p-ERK1/2, GSK3β, p-GSK3β, mTOR, and p-mTOR protein in lung adenocarcinoma and squamous cell carcinoma cells. RESULTS The correlation of Cbl-b expression and OS was different between NSCLC adenocarcinoma and squamous carcinoma. After transfection, the expression of Cbl-b was inhibited in A549, H1975, and SW900 cells. Cbl-b shRNA promoted the migration and invasion of lung adenocarcinoma A549 and H1975 cells, but it inhibited the invasion of lung squamous cell carcinoma SW900 cells. In addition, Cbl-b regulated the expression of PI3K and ERK1/2-GSK3β pathway proteins in A549 and SW900 cells. CONCLUSIONS The OS of Cbl-b mRNA low expression in lung adenocarcinoma and squamous cell carcinoma was different. The difference in signal pathways may be one of the reasons for the difference in the correlation between Cbl-b expression and the survival rate of these 2 pathological types of lung cancer.

灵芝入药在我国有着悠久的历史.灵芝所含的化学成分复杂,研究表明灵芝多糖是其主要的活性成分,具有广泛的药理作用.本文主要介绍灵芝多糖类化合物的现代药理学研究进展,为灵芝多糖的开发利用提供前沿情报参考.

目的 探讨玫瑰茄不同提取物对小鼠醉酒的预防作用.方法 采用预防醉酒实验,观察玫瑰茄不同提取物对小鼠醉酒的影响.结果玫瑰茄水提醇沉上清部分能明显缩短小鼠平均醉酒时长,与对照组相比,有显著性差异(P<0.05).结论 玫瑰茄水提醇沉上清部分对小鼠醉酒具有较显著的预防作用.

Radix et Rhizoma Sophorae tonkinensis is a traditional Chinese herb and endangered medical plant,which has rich medical functions and toxic effects so that it has been paid extensive attention.This article aims at reviewing the recent literatures on chemical constituents,pharmacological activities,toxicological effects as well as quality control so as to provide reference for its integrated exploitation and utilization and promote the reasonable combination of its chemical constituents,bioactivity,quality control with clinical application.

Objective To study the enhancement effect of paclitaxel (PTX) induced apoptosis in HER2+ breast cancer cells by the triterpene component of Ganoderma lucidum (GLE) and its mechanism . Methods Cell proliferation was analyzed by SRB and trypan blue exclusion staining method . The ex‐pression of proteins was detected by Western blot . Antitumor activity in vivo was studied in SKBR‐3 xenograft model . Results In vitro ,GLE and PTX had synergistic effect on the proliferation and induced apoptosis in SKBR‐3 cells ,and on the upregulation of proteins related to cell apoptosis and blocking trans‐duction of HER2 signaling pathway . In vivo ,GLE enhanced antitumor activity of PTX against HER2+breast cancer . Conclusion GLE and PTX have synergistic antitumor effect against the HER2+ breast cancer cells SKBR‐3 both in v itro and in v ivo .

Phytochemical studies on the ethyl acetate extract of the fruiting body of Ganoderma lucidum afforded a new lanostanoid named 24(S),26-dihydroxy-5 α -lanosta-7,9,25-trien-3-one ( 1 ), along with two known compounds, ganoderic acid DM ( 2 ) and 5 α -lanosta-7,9(11),24-triene-15 α ,26-dihydroxy-3-one ( 3 ). Their structures were established on the basis of spectroscopic analysis and chemical evidence. The cytotoxicity of the isolated compounds was evaluated in vitro against three human cancer cell lines (HCT-116, KB, and OE-19). Compound 3 showed moderate cytotoxic activity.

目的:观察灵芝三萜胶囊对动物神经、呼吸和心血管系统的影响.方法:灌胃给药后,旷野法观察灵芝三萜胶囊对小鼠自发活动的影响;金属棒法和倾斜板法观察灵芝三萜胶囊对小鼠机能协调功能的影响;采用入睡率和睡眠时间观察灵芝三萜胶囊与戊巴比妥钠联用对小鼠的睡眠等神经系统方面的影响;使用生理机能记录仪记录给药前后对麻醉大鼠的呼吸频率、血压、心率等呼吸、心血管系统方面的影响.结果:灵芝三萜胶囊(250～ 1000mg/kg)对小鼠的自主活动、机能协调功能、戊巴妥钠阁下催眠剂量及戊巴比妥钠睡眠时间均无明显的影响;灵芝三萜胶囊(125～500mg/kg)对麻醉大鼠的呼吸和心血管系统无影响.结论:灵芝三萜胶囊对动物的神经、呼吸和心血管系统均无明显的影响作用,在药效学有效剂量范围内有较高的安全性.