虎耳草及其组培快繁技术

综述了虎耳草的生物、生态学特性,用途,研究和开发利用现状等。组培繁殖试验结果表明,以虎耳草成熟植株的茎段为外植体,在无激素培养基MS上诱导出芽;在MS 6-BA0．2mg／L(单位下同) IBA0．1培养基上继代和增殖培养,繁殖系数可达4～5;在培养基1／2MS 15g/L糖的培养基上进行根系诱导及生长,生根率可达100％,开始生根时间3d,根系生长完成时间14～15d,生根整齐,同步性高。上述培养务件为本试验的最佳配方。虎耳草组培再生植株的移栽成活率可达85％以上。     

Effect of ethanol extract from Chrysanthemum cinerariifolium leaves on Ki-67 proliferation and dysplasia severity in a rat model of oral squamous cell carcinoma

Background:Oral squamous cell carcinoma (OSCC) is a malignant tumor that can rapidly infiltrate the oral epithelial tissue and cause high mortality worldwide because the available therapies are less effective. Chrysanthemum cinerariifolium leaf contains secondary metabolites as anti-inflammatory, antioxidant, anticancer, and antimutagenic. Aims:The study aimed to analyze the ethanolic extract of C. cinerariifolium leaf in reducing proliferation (Ki-67) and the degree of dysplasia in OSCC rats. Methods:This study used male Sprague Dawley induced by 7,12-dimethylbenz(a)anthracene (DMBA) 0.5% and divided into five treatment groups, namely positive control/C+ (sick), negative control/C- (healthy), and treatment group induced with DMBA and given extract C. cinerariifolium leaf with successive doses of T1, T2, and T3 (50, 100, and 200 mg/kg bw). The oral epithelium was stained with hematoxylin and eosin and immunohistochemically stained with a Ki-67 monoclonal antibody. The statistical analysis utilizes the one-way analysis of variance test. Results:The results showed that T1 at a dose of 200 mg/kg bw could significantly reduce Ki-67 expression and the degree of oral epithelial dysplasia (OED; p < 0.05) close to healthy controls. Conclusion:The conclusion shows that C. cinerariifolium leaf extract can be a therapy against OSCC by decreasing cell proliferation and the degree of OED.     

紫锥菊多糖对 IEC-6细胞增殖的影响

[目的]研究紫锥菊多糖对大鼠小肠上皮细胞株 IEC-6细胞增殖的影响。[方法]以IEC-6细胞为研究对象,采用 MTT方法检测细胞增殖率,观察紫锥菊多糖对该细胞增殖的影响。[结果]紫锥菊不同剂量与不同的培养时间对 IEC-6细胞的生长均有促进作用,但是随着处理时间的延长,不同浓度的促增殖效果也不尽相同,表现为在浓度为50、200μg/ml 时增殖率随着处理时间的延长先增加后逐渐减弱,但在培养24 h后表现显著促增殖作用;浓度100μg/ml 在72 h、浓度500μg/ml 在48 h表现明显促增殖作用;经48 h处理的EPS其增殖率随浓度提高而增强。[结论]紫锥菊多糖具有促进 IEC-6细胞增殖的作用,通过对小肠上皮细胞增殖的影响而发挥对肠道黏膜吸收和免疫功能的调整作用。     

In vitro embryo germination and proliferation of pistachio (Pistacia vera L.)

In vitro development of isolated embryos and axillary bud proliferation were studied in Pistacia vera L. Different regulator-free nutrient media were compared to determine the effects of the mineral solution, agar and sucrose concentrations on seedling development from mature embryos. Nutrient-rich MS [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tabacco tissue cultures. Physiol. Plant. 15, 473–479] and DKW [Driver, J.A., Kuniyuki, A.M., 1984. In vitro propagation of Paradox walnut rootstock. HortScience 19, 507–509] mineral solutions were more efficient for the development of aerial parts than nutrient-poor KN [Knop, W., 1884. Bereitung einer concentrierten nährstofflosung für pflanzen. Landwersuhssat 30, 292–294] and WT [Withe, P.R., 1936. Plant tissue cultures. Bot. Rev. 2, 419–437] solutions. Reducing the agar concentration enhanced fresh matter production and balanced seedling development. When tested at different concentrations, sucrose was found to orient mature embryo development, with the best results obtained at concentrations of 2–4%, whereas high concentrations (6 and 12%) mainly inhibited elongation of the aerial parts. Plantlets obtained previously from in vitro cultured embryos were propagated by axillary budding. High bud proliferation (six shoots per explant) was achieved when using 17.8 μM benzyladenine (BA) combined with 0.5 μM indole-3-butyric acid (IBA). The addition of 2.9 μM gibberellic acid (GA₃) to the propagation medium did not improve axillary shoot yields and on average, media with GA₃ produced 2.3–2.6 elongated stems per cultured explant. Shoots were rooted in vitro in half-strength MS medium containing 12.3 μM IBA.     

Knock-down of NEDD1 expression inhibits lung adenocarcinoma cell proliferation and migration

AIM: To explore the role of neural precursor cell expression developmentally down-regulated protein 1 (NEDD1) in the development and progression of lung cancer. METHODS: The differences of NEDD1 expression levels between lung cancer tissues and tumor-adjacent tissues were analyzed by the method of immunohistochemistry and TCGA database. Kaplan-Meier curve was used to analyze the correlation between lung cancer prognosis and the expression level of NEDD1. The proliferation of A549 cells was tested by plate colony formation experiment after knock-down of NEDD1 expression. The apoptosis rate and cell cycle distribution were examined by flow cytometry. The migration ability of the A549 cells was detected by Transwell assay. The protein levels of cell cycle-related molecules were determined by Western blot. Database analysis was performed to evaluate the relationship between the expression of NEDD1 and cyclin-dependent kinases 2 (CDK2). RESULTS: Compared with the tumor-adjacent tissues, the expression level of NEDD1 in the lung cancer tissues was increased, so as the database analysis, and the higher expression of NEDD1 showed a poorer prognosis. Under light microscope, the A549 cells showed a low proliferation rate after silencing the NEDD1 expression, and the colony formation ability of the cells was also reduced; knock-down of NEDD1 expression induced the apoptosis and inhibited the cell migration; knock-down of NEDD1 expression blocked the cells in G₁/S phase, and the protein levels of p-Rb and cyclinD1 were decreased, while the protein levels of p-Chk1, p-Chk2 and p-p53 were increased (P<0.05). A positive correlation between the expression of NEDD1 and CDK2 was noted by database analysis. CONCLUSION: NEDD1 plays an crucial role in promoting cell proliferation via inhibiting apoptosis and accelerating cell cycle, high expression of NEDD1 in lung adenocarcinoma tissue is related to poor prognosis, thus NEDD1 may be used as a candidate marker molecule for the diagnosis and prognosis of lung cancer.     

N-糖基化对乙型脑炎病毒感染与增殖的影响

N-糖基化对于病毒的感染与增殖中均具有重要作用,切除病毒表面糖链或抑制病毒糖基化修饰的方法已成为登革热病毒等的潜在治疗方法.为阐明日本乙型脑炎病毒(Japanese encephalitis virus,JEV)表面的糖链在其感染和增殖过程中的作用,本研究用肽N-糖苷酶F(peptide N-glycosidase F,PNGase F)和内切糖苷酶H(endoglycosidase H,Endo H)处理乙型脑炎病毒,将酶切后的病毒用Western blot验证糖基化酶切的效果,将酶切的JEV接种BHK-21细胞进行蚀斑试验,观察蚀斑的大小及病毒滴度的变化情况.同时用衣霉素处理BHK-21细胞并同步接种JEV,进行蚀斑试验,观察蚀斑大小的变化与病毒增殖情况.结果显示,用Endo H和PNGase F处理JEV后,病毒形成的蚀斑均显著小于未处理的病毒对照,且病毒增殖滴度显著降低,说明病毒粒子表面的糖基化修饰对于病毒的侵染具有重要作用.衣霉素处理细胞后,病毒产生的蚀斑大小没有明显差异,但增殖滴度却显著降低,说明糖基化在JEV增殖过程中也具有重要的功能.本研究说明JEV的糖基化对于其感染和增殖均具有重要的作用,为开发抑制乙型脑炎病毒药物研究提供一个新的思路.     

兔眼蓝莓组织培养与快繁技术研究

[目的]研究兔眼蓝莓的组织培养与快繁技术。[方法]以蓝莓品种的幼嫩茎段为试材,研究不同取材时间、不同品种对茎段诱导效果及不同培养基及激素组合对灿烂继代增殖及生长的影响,并开展瓶外驯化生根试验。[结果]4月下旬到6月上旬为蓝莓茎段最佳取材时期,萌芽率为62.2%;灿烂品种在改良WPM+2.0 mg/L ZT+0.05 mg/L IBA+30 g/L蔗糖+6.5 g/L琼脂培养基中生长最好,茎段增殖率可达3.21;继代40～50 d的茎段用1 000 mg/L ABT快速浸蘸处理后扦插于装有苔癣的穴盘上,生根率达85.33%;生根组培苗在草炭∶园土∶珍珠岩=2∶1∶0.5的基质中移栽成活率为85.67%。[结论]为蓝莓的南方适栽品种的繁殖提供了技术支持。     

蝴蝶兰类原球茎的诱导

以蝴蝶兰优良杂交组合NoⅡ(♀)×No2(♂)和优良品种EA2003无菌苗的叶片为试材,对影响类原球茎(PLB)的发生、增殖等因子进行系统研究。结果表明:1/4MS+6-BA10 mg·L-1+椰子汁100 g·L-1是蝴蝶兰PLB诱导最适宜的培养基;100 g·L-1椰子汁对PLB诱导有明显的促进作用;适宜PLB增殖的培养基是1/4MS+2,4-D1.0 mg·L-1+6-BA2.0 mg·L-1;活性炭可以有效防止NoⅡ(♀)×No2(♂)叶片的褐化死亡,但活性炭对蝴蝶兰优良品种EA2003 PLB的诱导有阻碍作用;叶片基部PLB的发生优于叶片的其他部位。