Targeting colorectal cancer with Herba Patriniae and Coix seed:Network pharmacology,molecular docking,and in vitro validation

BACKGROUND Herba Patriniae and Coix seed(HC)constitute a widely utilized drug combination in the clinical management of colorectal cancer(CRC)that is known for its diuretic,anti-inflammatory,and swelling-reducing properties.Although its efficacy has been demonstrated in a clinical setting,the active compounds and their mechanisms of action in CRC treatment remain to be fully elucidated.AIM To identify the active,CRC-targeting components of HC and to elucidate the mechanisms of action involved.METHODS Active HC components were identified and screened using databases.Targets for each component were predicted.CRC-related targets were obtained from human gene databases.Interaction targets between HC and CRC were identified.A“drug-ingredient-target”network was created to identify the core components and targets involved.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses were conducted to elucidate the key pathways involved.Molecular docking between core targets and key components was executed.In vitro experiments validated core monomers.RESULTS Nineteen active components of HC were identified,with acacetin as the primary active compound.The predictive analysis identified 454 targets of the active compounds in HC.Intersection mapping with 2685 CRC-related targets yielded 171 intervention targets,including 30 core targets.GO and KEGG analyses indicated that HC may influence the phosphoinositide 3-kinase(PI3K)/Akt signaling pathway.Molecular docking showed that acacetin exhibited an optimal interaction with AKT1,identifying PI3K,AKT,and P53 as key genes likely targeted by HC during CRC treatment.Acacetin inhibited HT-29 cell proliferation and migration,as well as promoted apoptosis,in vitro.Western blotting analysis revealed increased p53 and cleaved caspase-3 expression and decreased levels of p-PI3K,p-Akt,and survivin,which likely contributed to CRC apoptosis.CONCLUSION Acacetin,the principal active compound in the HC pair,inhibited the proliferation and migration of HT-29 cells and promoted apoptosis through the PI3K/Akt/p53 signaling pathway.

Two New Antibacterial Iridoids from Patrinia rupestris

Two new iridoids （1 and 2） were isolated from the roots of Patrinia rupestris （Pail.） Juss. Their structures were elucidated by spectroscopic methods. Compounds 1 and 2 exhibited strong antibacterial activities against Eschecichia coli and Staphylococcus aureus, respectively.

Lignans from Patrinia scabiosaefolia improve insulin resistance by activating PI-3K/AKT pathway and promoting GLUT4 expression

Patrinia scabiosaefolia,is used as wild vegetable in China for more than 2000 years,with a variety of pharmacological activities,including anti-inflammatory,anti-tumor and hypoglycemic.Based on our ongoing research on chemical constituents and hypoglycemic activity of P.scabiosaefolia,4 lignan compounds,(+)-isolariciresinol(1),7R,7’R,8S,8’S-(+)-neo-olivil-4-O-β-D-glucopyranoside(2),4-O-methylcedrusin(3)and patrinian A(4),were isolated and identifi ed.The hypoglycemic activity showed that compounds 2 and 3 could extremely signifi cantly improve insulin resistance at 100(P<0.001),50(P<0.001)and 25μmol/L(P<0.01)in IR 3T3-L1 cells.While compound 4 only promoted glucose uptake by IR 3T3-L1 cells at 100μmol/L(P<0.01).Western blotting experiments showed that compounds 2 and 4 up-regulated the protein expressions of p-IRS,PI-3K,p-AKT and glucose transporter 4(GLUT4),and promoted the transcription of GLUT4 mRNA.Therefore,the mechanisms of compounds 2 and 4 were presumed to improve IR by activating PI-3K/AKT signaling pathway.

岩鸽排泄物对壁画损害特征研究

鸟类排泄物是造成石窟寺壁画、彩塑污染的主要因素之一,科学剖析鸟类排泄物对壁画的损害特征是此类污染物有效清除的前提。本研究利用傅里叶变换红外光谱(FTIR)、气相色谱-质谱联用(GC-MS)、X射线衍射(XRD)等方法分析了炳灵寺石窟壁画表面岩鸽排泄物成分,采用扫描电镜-能谱仪(SEM-EDS)、便携式显微镜、偏光显微镜(PLM)表征了鸟类排泄物对模拟壁画的微观损害特征。结果表明,岩鸽排泄物成分主要由尿酸、纤维素、多肽类有机物,以及以石英为主的无机盐组成。排泄物造成壁画覆盖污染、颜料层和地仗层侵蚀、颜料颗粒间原始粘附结构破坏,导致颜料层起甲、脱落,可溶盐活动及其他生物侵蚀。研究结果对于古代壁画的科学保护与修复具有指导意义。

败酱草水提物通过调控circ_0000936/miR-665抑制胶质瘤细胞增殖、迁移及侵袭

目的:探讨败酱草水提物对胶质瘤细胞生物行为的影响及其可能作用机制。方法:收集2019年2月至2020年1月期间于本院接受手术切除并经病理证实的31例胶质瘤组织,qRT-PCR法检测正常脑组织与胶质瘤组织中circ_0000936、miR-665的表达量;Pearson法分析胶质瘤组织中circ_0000936和miR-665表达量的相关性;体外培养人胶质瘤细胞LN229,不同浓度的败酱草(0.5 mg/mL、1.0 mg/mL、2.0 mg/mL)处理LN229细胞,si-NC、si-circ_0000936分别转染至LN229细胞,pcDNA-circ_0000936转染至LN229细胞后加入2.0 mg/mL败酱草处理细胞;CCK-8、平板克隆形成实验、Transwell小室实验与流式细胞术分别检测细胞增殖、克隆形成、迁移、侵袭及凋亡能力;双荧光素酶报告基因实验检测circ_0000936与miR-665的靶向关系;Western blot法检测Bax、Bcl-2蛋白表达量。结果:胶质瘤组织中circ_0000936的表达量高于正常脑组织(P<0.05),而miR-665的表达量低于正常脑组织(P<0.05);circ_0000936和miR-665呈负相关(r=-0.980,P<0.001);败酱草能够以浓度依赖性方式降低细胞活力、细胞克隆形成率、Bcl-2蛋白水平和circ_0000936的表达量(P<0.05),迁移及侵袭细胞数减少(P<0.05),而凋亡率、Bax蛋白水平和miR-665的表达量升高(P<0.05);转染si-circ_0000936后miR-665的表达量升高(P<0.05),细胞活力、细胞克隆形成率和Bcl-2蛋白水平降低(P<0.05),迁移及侵袭细胞数减少(P<0.05),凋亡率和Bax蛋白水平升高(P<0.05);转染pcDNA-circ_0000936能够逆转败酱草对LN229细胞生物学行为的作用。结论:败酱草水提物可通过调控circ_0000936/miR-665诱导胶质瘤细胞凋亡,并阻碍增殖、迁移及侵袭。

新疆一枝蒿不同部位水提物的体外抗炎效果

为了探究新疆一枝蒿不同部位水提物的体外抗炎效果,本试验在体外用脂多糖(LPS)诱导小鼠单核巨噬白血病细胞(RAW264.7)构建炎症模型,采用水提法对新疆一枝蒿不同部位(叶、茎、花)进行提取,运用噻唑蓝(MTT)法检测新疆一枝蒿不同部位水提物对细胞活力度的影响,实时荧光定量PCR(RT-qPCR)检测其对炎症相关基因TNF-α、IL-1β和IL-6 mRNA表达的影响,Western blot检测其对TNF-α、IL-1β和IL-6蛋白表达的影响。结果显示,新疆一枝蒿叶、茎、花水提物分别在0.078~1.250、0.078~5.000和0.078~1.250 mg/mL的浓度范围内对RAW264.7细胞活力度无显著抑制作用。与LPS模型组相比,新疆一枝蒿叶、茎、花水提物浓度范围在0.313~1.250、0.625~2.500和0.625~1.250 mg/mL时,细胞活力度分别提高了23.12%~28.14%(P<0.01)、10.88%~18.62%(P<0.01)和16.13%~16.76%(P<0.05)。浓度为1.250 mg/mL的叶水提物可以极显著降低TNF-α、IL-1β和IL-6的mRNA和蛋白表达(P<0.01);浓度为0.625和1.250 mg/mL的茎水提物可以极显著降低IL-1β的mRNA和蛋白表达(P<0.01),以及IL-6和TNF-α的mRNA表达(P<0.01);浓度为0.625 mg/mL的花水提物可以极显著降低IL-1β和IL-6的mRNA和蛋白表达(P<0.01)。结果表明,新疆一枝蒿不同部位(叶、茎、花)水提物均有抗炎效果,本试验结果可为新疆一枝蒿的兽医临床应用提供参考依据。

pH-时滞型新疆一枝蒿结肠定位片的制备及体外释放评价

目的:制备pH-时滞型新疆一枝蒿结肠定位片并对其体外释放性能进行评价。方法:采用压制包衣、喷雾包衣的方法制备结肠定位片,以总黄酮累积释放度为指标,分别考察压制包衣时滞材料处方比例、包衣用量、上下层填料比例,pH控释层处方比例与包衣增重。结果:经过考察,最优工艺处方为EC-PEG 6 000(4∶1),压制包衣0.1 g,压制包衣上下层填料(1.5∶1),Eudragit FS 30 D-Eudragit L30 D-55(2∶1),p H控释层包衣增重为7%时,制得结肠定位制剂在0.1 mol/L HCl溶液中2 h内不释药,在pH 6.8磷酸盐缓冲液中4 h累积释放度<10%,在p H 7.5磷酸盐缓冲液中6 h累积释放度>80%。结论:经过筛选后的结肠定位片能够实现结肠定位释放的目的且体外释放稳定。

新疆一枝蒿口服液的制备工艺优化研究

为了探究新疆一枝蒿口服液的最佳制备工艺,采用水提醇沉法提取新疆一枝蒿中的总黄酮,经单因素、响应面试验获取最佳的制备工艺参数,并通过薄层层析与含量限度测定对新疆一枝蒿口服液建立初步的质量控制。结果表明:新疆一枝蒿最佳制备工艺为料液比1∶14,煎煮时间为1.5 h,醇沉浓度为50%。供试品色谱中,在与对照品色谱相应的位置上,显相同颜色的斑点,含量限度测定黄酮含量范围在28.67~29.59 mg/mL,平均值为29.10 mg/mL,最低限度为23.28 mg/mL。

白花败酱草化学成分与药理活性研究进展

白花败酱草是败酱属植物,分布广泛,植物资源量大。在我国,白花败酱草具有悠久的药用历史,作为一味传统中药在多种方剂中出现。在中医临床应用中,白花败酱草在治疗呼吸系统、消化系统、泌尿生殖系统、妇科、皮肤科等多学科疾病的治疗实践中得到了广泛应用。目前,对于白花败酱草的化学成分研究进展颇丰,已提取分离出81种单体成分,包括苯丙酸类、黄酮类、萜类等多种结构类型。同时,药理学研究发现白花败酱草具有多种明确的药理活性,包括抗肿瘤、消炎镇痛、抑菌以及保护肝脏等,并对其作用机制进行了初步研究探讨。该文在临床功效、化学成分、药理作用等方面对白花败酱草进行了综述,归纳总结现有已发表的研究成果,为其临床应用和进一步研究提供依据和基础。

败酱草质量研究

目的:建立败酱草的薄层鉴别方法(TLC)及HPLC测定东莨菪内酯含量测定方法,为败酱草的质量研究提供依据。方法:采用TLC鉴别法,HPLC作为含量测定方法,色谱柱:Ultimate Plus C_(18)(4.6 mm×250 mm,5μm);流动相:乙腈-0.5%冰醋酸(15∶85);柱温:25℃;流速:1 mL·min^(-1);检测波长:343 nm。结果:薄层鉴别中检出与对照药材相同颜色的荧光斑点;东莨菪内酯在0.8384~20.96μg·mL^(-1)范围内线性关系良好(函数关系为Y=39.233X+7.0215,r=0.9999),回收率为91.0%~97.9%,均值为95.4%,RSD(n=6)为3%。结论:本方法鉴别专属性较好,含量测定方法专属性、稳定性、重现性、准确度良好,可用于败酱草的质量控制。

A New Alkaloid from Patrinia scabra

A new alkaloid has been isolated from the root of Patrinia scabra. Its structure was elucidated as 2'-acetamido-3'-phenyl propyl 2-benzamido-3-phenyl propionate by extensive spectroscopic analysis.

Two New Dihydroflavanoids from Patrinia villosa Juss

Two new dihydroflavonoids were obtained from the traditional Chinese medicinal herb Patrinia villosa Juss. Their structures were elucidated as （2S）-5, 7, 2＇, 6＇-tetrahydroxy-6, 8-di （γ, γ-dimethylallyl） flavanone （1） and （2S）-5, 7, 2＇, 6＇-tetrahydroxy-6-1avandulylated flavanone （2） by spectroscopic methods including UV, IR, HR-TOF-MS, 1D NMR and 2D NMR techniques.

Inhibitory effect of Patrinia scabiosaefolia on acute pancreatitis

瞄准：在缩胆囊素(CCK ) 上调查 Patrinia scabiosaefolia (PS ) 的效果在老鼠的导致 octapeptide 的尖锐胰腺炎(AP ) 。方法：称 240-260 g 的 Wistar 老鼠被划分成三个组：(1 ) 正常的盐对待的组；(2 ) 在哪个 PS 口头上地被管理，有在 100 mg/kg 组的 PS 的处理在 1， 3 和 5 h，和这个全部以后由 75 microg/kg CCK octapeptide 的下的管理列在后面三次过程为 5 d 被重复；(3 ) 与盐的组，协议与在处理一样在是一起的处理与 PS 组织。我们决定了胰腺的重量 / 身体重量比率，胰腺的 HSP60 的层次， HSP72 和支持 inflammatory cytokines 的分泌物。重复 CCK octapeptide 处理导致了试验性地导致的胰腺炎的典型实验室调查结果。结果：PS 在 CCK 导致 octapeptide 的 AP 减少了胰腺的重量 / 身体重量比率，浆液淀粉酶和脂肪分解酵素的层次，和支持 inflammatory cytokines 的禁止的表情。而且， PS 预告的处理增加了 HSP60 和 HSP72 的胰腺的层次。结论：有 PS 的预告的处理在 CCK 导致 octapeptide 的 AP 上有反煽动性的效果。

Differences in Morphological, Physiological and Growth Traits between Two Endemic Subspecies of <i>Brassica rupestris</i>Raf.: Implications for Their Conservation

The goal of this work was to analyze the phenological and physiological responses of Brassica rupestris Raf. subsp. hispida Raimondo & Mazzola and Brassica rupestris Raf. subsp. rupestris to environmental factors also in consideration of global change. We used seedlings of the two subspecies originated from wild plants from the natural habitats in Sicily and cultivated in the Botanical Garden of Rome. Leaf morphological and physiological traits and growth dynamic were analyzed as well as the response to an imposed water stress experiment. The results underlined a higher relative growth rate in plant height (RGRH, cm·cm-1·d-1) in B. rupestris subsp. rupestris also attested by the highest plant height (H, cm), leaf area (LA, cm2), specific leaf area (SLA, cm·g-1), total leaf area per plant (TLA, cm2) and total plant dry mass (PDM, g plant-1). Moreover, the significantly highest net photosynthetic rates (A, μmol CO2 m-2·s-1) during the study period of this subspecies was related to the high Chlorophyll content (Chl, SPAD units). B. rupestris subsp. rupestris was also the most tolerant subspecies to imposed water stress showing the highest relative water content (RWC, %), A and water use efficiency (WUE, μmol·mmol-1).

Two New Dihydroflavanoids from Patrinia vittosa Juss (II)

Two new dihydroflavonoids named villosin A and viUosin B were obtained from the traditional Chinese medicinal herb Patrinia viUosa Juss. Their structures were elucidated as （2S）-5, 2＇, 6＇-trihydroxy-2＂, 2＂-dimethylpyrano [5＂, 6＂： 6, 7]flavanone 1 and （2S, 3＂S）-5, 2＇, 6＇- trihydroxy-3＂-γ, γ-dimethylallyl-2＂, 2＂-dimethyl-3＂, 4＂-dihydropyrano [5＂, 6＂： 6, 7] flavanone 2 by spectroscopic methods including UV, IR, HR-TOF- MS, 1D NMR and 2D NMR techniques.

Scabrous patrinia root inhibits circulating tumor cells in differentiated thyroid carcinoma–a clinical observation

Objective To observe the clinical inhibition of circulating tumor cells(CTCs) in differentiated thyroid carcinoma(DTC) by the extract of scabrous patrinia root(Huikangling).Methods Eighty-seven DTC patients tested positive for CTCs were randomly divided into two groups; all patients were treated with oral levothyroxine sodium in accordance with the DTC endocrine inhibition treatment criteria. Patients(n = 45) in the treatment group were provided the standard endocrine therapy along with oral Huikangling(0.4 g/tablet, 0.4 g × 3/time, 3 times/day, 12 weeks). Patients(n = 42) in the control group were only provided the standard therapy. After 4 and 12 weeks, CTCs in the blood were detected by flow cytometry.Results After 4 weeks of oral Huikangling treatment, CTCs were detected in 18(40%) and 29(69%) patients in the treatment and control groups, respectively; the difference was statistically significant(χ2 = 8.49, P < 0.05). After 12 weeks, CTCs were detected in 7(15.6%) and 17(44.7%) patients in the treatment and control groups, respectively; the difference was statistically significant(χ2 = 5.68, P < 0.05). Follow-up evaluation revealed two patients with lung metastasis and one patient with bone metastasis in the control group; one patient showed lateral neck lymph node metastasis without local recurrence in the treatment group.Conclusion Huikangling treatment reduces the number of CTC-positive DTC cases; however, further studies are needed to elucidate the underlying mechanisms.

Elimination of grapevine fleck virus and grapevine rupestris stem pitting-associated virus from Vitis vinifera 87-1 by ribavirin combined with thermotherapy

Vitis vinifera 87-1 plants infected by grapevine fleck virus(GFkV) and grapevine rupestris stem pitting-associated virus(GRSPaV) were used as the plant materials for virus elimination treatment. This study evaluated the effects of ribavirin at different concentrations(15 and 25 μg mL^(–1);R15 and R25, respectively), thermotherapy(37°C;T), and the combination of ribavirin and thermotherapy(R15+T and R25+T) on eliminating viruses from grapevine plants in vitro. Both R15 and R25 had phytotoxic effects and weakened plant growth. Thermotherapy positively affected the growth of grapevine plants. Plant height was significantly greater in T, R15+T, and R25+T than in CK, R15 and R25. The proportion of dead plants after T, R15+T, and R25+T was 51.4, 11.4, and 8.6%, respectively. The survival rates of regenerated plants after all treatments were >68.0%. Ribavirin concentration and treatment time were related to the regeneration of shoot tips and elimination efficiencies of the two viruses. The survival rates of plants after R15+T for 30, 40, and 50 days were 97.3, 90.7, and 74.4%, respectively. The elimination rates of GRSPaV from plants in the three time quantum were 55.6, 84.6, and 93.8%, respectively. The elimination rate of GFkV was 23.9% higher in R25(35/44) than in R15(25/45), and that of GRSPaV was 7.0% higher in R25 than in R15. The combination of thermotherapy and chemotherapy was found to have a positive effect on the eradication of GFkV and GRSPaV, and R25+T for 50 days was able to completely eliminate the two viruses from in vitro grapevines.

Development of a Total Flavonoids Extract of Artemisia Rupestris L.via Nanotechnology and Its Antiviral Effect In Vitro

Objective:We aimed to develop a novel Artemisia rupestris L.flavonoid nano-encapsulation(AFN)preparation and evaulate its anti-hepatitis B virus(HBV)activityin vitro.Methods:First,the AFN was prepared using polylactic-co-glycolic acid(PLGA).Then,after verification of the AFN,in vitro anti-virus assays were conducted by:(1)assessing the inhibitory effect of AFN on the secretion of hepatitis B surface antigens(HBsAg),hepatitis B e-antigens(HBeAg),and the replication of HBV DNAin HepG2.2.15 cells;(2)analyzing the influence of AFN on the activation rate of NF-κB positive cells;and(3)evaluating the effect of AFN on the function of glutathione peroxidase(GSH-PX)enzymes located onthe HepG2.2.15 cell membrane.Results:Compared to the original total flavonoids extract of Artemisia rupestris L.(withoutnano-encapsulation),AFN preparation under the maximum nontoxic concentration effectively inhibited the secretion of HBsAg,HBeAg,and HBV DNA from HepG2.2.15 cells.At the same time,AFN preparation promoted not only the activation rate of NF-κB positive cells,but also antiviral GSH-PX enzyme function.In conclusion,nano-encapsulation of the flavonoids extract of Artemisia rupestris L.showed an enhanced anti-HBV effect in vitro compared to the original total flavonoids extract(without nanoencapsulation);therefore,nano-encapsulation has great potential for the development of a novel antiviral herbal medicine preparation with improved efficacy.

Network pharmacology-based screening of the active ingredients and potential targets of the Patrinia villosa for application to Alzheimer’s disease

Alzheimer’s disease(AD)is a progressive neurodegenerative disorder associated with global deterioration of intellectual function,including memory,learning,orientation,language,comprehension and judgment.Given the prevalence of AD and the lack of effective long-term therapies,there is a pressing need to discover viable leads that can be developed into clinically approved drugs with disease-modifying effects.The analysis of current reported literatures confirms the importance of Patrinia villosa J.as candidate against AD.Hence,the compound-disease target network based on the methods of network pharmacology prediction was established in this paper.The aim is to explore potential compounds with marked effect on AD in P.villosa.The result showed that the most effective compound in this plant might be threo-(7R,8R)-guaiacyl-glycerol-β-O-4′-dihydroconiferyl ether,and the most potential target might be Maltose-binding periplasmic.

栽培一枝蒿粗多糖混合口蹄疫疫苗乳化方法及稳定性分析

旨在探究栽培一枝蒿粗多糖(cultivated Artemisia rupestris L.crude polysaccharides,CARCP)协同ISA-206油乳剂通过比较不同乳化方法与口蹄疫灭活抗原(inactivated foot-and-mouth disease viral antigen,FMD-Ag)制备FMD疫苗的效果异同,并观察不同储存条件对FMD疫苗稳定性的影响。通过搅拌和超声乳化分别制备不同疫苗组合,皮下免疫ICR小鼠后检测FMD特异性抗体及T细胞亚群比例。将搅拌乳化的不同疫苗组合在4、25℃条件下放置30、180 d后免疫ICR小鼠,检测体液和细胞免疫水平分析其稳定性。结果显示:与商品化FMD疫苗相比,搅拌和超声乳化制备的CARCP单独或协同ISA-206油乳剂的FMD疫苗组的特异性IgG水平和CD3^(+)CD4^(+)、CD3^(+)CD8^(+)T细胞比例无显著性影响(P>0.05)。4℃放置30、180 d后,与商品化FMD疫苗相比,CARCP单独或协同ISA-206油乳剂的FMD疫苗组的IgG水平和CD4^(+)、CD8^(+)、CD4^(+)CD44^(+)、CD8^(+)CD44^(+)T细胞比例无显著性差异(P>0.05)。25℃放置30、180 d后,与商品化FMD疫苗相比,免疫动物后FMD-Ag+ISA-206抗体水平及T细胞亚群比例显著降低(P<0.05),而FMD-Ag+ISA-206+CARCP抗体水平及T细胞亚群比例无显著性差异(P>0.05)。搅拌和超声乳化制备的不同FMD疫苗组合的抗体水平和T细胞水平的免疫效果没有差异;不同储存条件下,CARCP能够增强FMD疫苗的稳定性,这些研究结果为CARCP多糖佐剂的研究提供试验依据。