螺旋藻多糖抗柯萨奇B3病毒的体外实验研究

在体外进行了钝顶螺旋藻多糖(polysaccharides from Spirulina platensis,PSP)抗柯萨奇病毒B3(Coxsackie virus B3,CVB3)活性的研究.以不同剂量的PSP作用于病毒复制周期的各个环节,以病毒半数感染量(TCID50),细胞病变效应(CPE),蚀斑形成单位(PFU),MTT染色细胞保护率(MTT法)作为评价指标,判断PSP的抗病毒效果.结果表明PSP对Vero细胞毒性极低(TC50为1750μg/mL),对CVB3无直接灭活作用,PSP可干扰病毒向宿主细胞的吸附,其抗病毒生物合成的活性最高(IC50为5.26,TI为237.6),是阳性对照药病毒唑的3.95倍,但不影响病毒的释放.提示PSP抗病毒靶位可能在于抑制病毒吸附和感染细胞内病毒的生物合成.     

一株红树林真菌抗单纯疱疹病毒Ⅱ型的研究

目的:对分离自海南红树林真菌菌株PH0016的胞外多糖(exopolysaccharide,EPS)抗单纯疱疹病毒Ⅱ型(Herpes simplex virus type 2,HSV-2)活性进行研究。方法:收集纯化菌株PH0016产生的EPS,体外采用细胞病变观察EPS的细胞毒性和抗HSV-2作用。为观察EPS对HSV-2体内感染的治疗效果,小鼠颅内注射0.02ml滴度为100LD50/0.1ml-1的HSV-2,24小时后以不同浓度EPS灌胃,持续7天,14天后计算小鼠存活率和存活时间。菌株分类采用形态学观察和ITS序列分析。结果:EPS可抑制HSV-2病毒活性,病毒感染的细胞病变的半抑制浓度(Inhibited concentration,IC50)为313μg/mL,SI为11.43。EPS 25mg.kg-1.d-1灌胃后,小鼠存活率为40.0%,存活时间为9.82±1.87天,相比模型组实验动物存活时间和存活率均有提高。菌株初步鉴定为淡紫色拟青霉。结论:从海南红树林分离一株淡紫色拟青霉,其产生的EPS具有抗HSV-2活性,值得进一步研究。     

青蒿提取物抗单纯疱疹病毒活性研究

用细胞病变效应(CPE)法证明了青蒿水提物具有抗单纯疱疹病毒Ⅱ型(HSV—2)活性,通过初步分离纯化得到抗HSV—2活性的有效成分。用MTT法研究了青蒿水提物和有效成分的细胞毒性和抗HSV—2活性,CC50分别为5．29mg／ml和4,94mg／ml,IC50分别为1．45mg／ml和0．128mg／ml,TI分别为3．65和38．6。以0．5mg／ml的无环鸟苷(ACV)作为阳性对照,结果显示有效成分在体外可以明显抑制HSV—2的致细胞病变作用,效果与ACV相当。     

灵芝多糖GLP的抗疱疹病毒作用机理

从灵芝菌丝体中分离得到的多糖GLP具有抑制单纯疱疹病毒I型感染的作用,并对GLP抑制疱疹病毒复制的作用机制进行了初步探讨。GLP对Vero细胞的CC50值大于2000μg/mL,GLP在疱疹病毒感染细胞前、感染后和病毒感染细胞时加入到细胞悬液中的EC50分别为4.6、50和17μg/mL;而如果将GLP与细胞共同孵育后再用病毒去感染,其EC50为11μg/mL。同时,GLP抑制病毒感染的选择指数分别为435、40、118和182。如果在病毒感染细胞后加入GLP,在病毒的生物大分子的合成完成后而子代病毒粒子还未释放出来前去掉GLP,这时GLP对病毒感染就没有抑制作用。定量PCR试验进一步证明GLP发挥抑制疱疹病毒感染作用是通过阻断病毒感染细胞早期与细胞表明蛋白的吸附来实现的。     

灵芝多糖GLP的抗疱疹病毒作用机理

从灵芝菌丝体中分离得到的多糖GLP具有抑制单纯疱疹病毒Ⅰ型感染的作用,并对GLP抑制疱疹病毒复制的作用机制进行了初步探讨.GLP对Vero细胞的CC50值大于2 000μg/mL,GLP在疱疹病毒感染细胞前、感染后和病毒感染细胞时加入到细胞悬液中的EC50分别为4.6、50和17μg/mL;而如果将GLP与细胞共同孵育后再用病毒去感染,其EC50为11μg/mL.同时,GLP抑制病毒感染的选择指数分别为435、40、118和182.如果在病毒感染细胞后加入GLP,在病毒的生物大分子的合成完成后而子代病毒粒子还未释放出来前去掉GLP,这时GLP对病毒感染就没有抑制作用.定量PCR试验进一步证明GLP发挥抑制疱疹病毒感染作用是通过阻断病毒感染细胞早期与细胞表明蛋白的吸附来实现的.     

生命泉膏滋抗实验小鼠阴道单纯疱疹病毒2型感染的研究

目的：研究中药生命泉膏滋（FESOL）抗小鼠阴道HSV－2感染的药效学作用,方法：以不同剂量FESOL作用于阴道感染HSV－2实验小鼠,观察用药后动物的发病率,死亡率和病变发展速度及程度。结果：FESOL高,中剂量组,[8.33,1.67,0.34g/(kg.d)]能显著降低感染鼠的发病率,死亡率,同时显著减轻病变程度,减慢病变发展速度。结论：中药FESOL能有效抗小鼠阴道HSV－2感染。     

甘草甜素抗单纯疱疹病毒I的实验研究

目的旨在观察甘草甜素对HSV-1的抑制作用,为临床更为有效的治疗单纯疱疹病毒性脑炎探索新的药物。方法分体外及体内实验两个部分。体外培养Vero细胞,在病毒吸附Vero细胞后加入甘草甜素,观察空斑形成单位来评价甘草甜素对HSV-1的抑制作用;建立单纯疱疹病毒性脑炎小鼠模型,于小鼠腹腔内给药。通过观察实验组和对照组的小鼠死亡率来评价甘草甜素在体内对HSV-1的抑制作用。结果甘草甜素能明显抑制病毒所致的空斑形成,IC50为0．56mM;实验组小鼠的死亡率明显低于对照组。结论甘草甜素能明显的抑制单纯疱疹病毒I型的复制,能明显降低单纯疱疹病毒性脑炎小鼠的死亡率。     

甘草甜素抗单纯疱疹病毒Ⅰ的实验研究

目的 旨在观察甘草甜素对HSV-1的抑制作用,为临床更为有效的治疗单纯疱疹病毒性脑炎探索新的药物.方法 分体外及体内实验两个部分.体外培养Vero细胞,在病毒吸附Vero细胞后加入甘草甜素,观察空斑形成单位来评价甘草甜素对HSV-1的抑制作用;建立单纯疱疹病毒性脑炎小鼠模型,于小鼠腹腔内给药.通过观察实验组和对照组的小鼠死亡率来评价甘草甜素在体内对HSV-1的抑制作用.结果 甘草甜素能明显抑制病毒所致的空斑形成,IC50为0.56 mM;实验组小鼠的死亡率明显低于对照组.结论 甘草甜素能明显的抑制单纯疱疹病毒Ⅰ型的复制,能明显降低单纯疱疹病毒性脑炎小鼠的死亡率.     

抗单纯疱疹病毒的天然产物研究现况

单纯疱疹病毒(HSV)是一种发病率极高,在全世界分布广泛的疱疹病毒,易引起多种人类皮肤疾病,严重危害人类的身体健康和生命安全。目前临床上常用核苷类抗病毒药物对单纯疱疹病毒感染进行治疗,但存在着许多明显的缺点,如：生物利用度低,溶解度小,临床多见不良反应且容易导致病毒耐药等。与核苷抗病毒药物相比,天然产物具有作用时间长、低毒、病毒不易产生耐药性等显著优点。随着对天然产物研究的深入,大量研究和实验证明：从植物或动物中提取的天然产物对于单纯疱疹病毒感染的治疗有着广阔的前景,天然产物成了寻找新型抗疱疹病毒药物的重要来源。将对近年来天然产物抗HSV研究状况进行整理、总结和分析,为将来天然产物抗HSV感染研究提供参考和对比。     

Application of Arthrospira (Spirulina) platensis biomass for silver removal from aqueous solutions

The cyanobacterium Arthrospira (Spirulina) platensis was used to study the process of silver biosorption. Effects of various parameters such as contact time, dosage of biosorbent, initial pH, temperature, and initial concentration of Ag(I) were investigated for a batch adsorption system. The optimal biosorption conditions were determined as pH 5.0, biosorbent dosage of 0.4 g, and initial silver concentration of 30 mg/L. Equilibrium adsorption data were analyzed by the Langmuir and Freundlich models – however, the Freundlich model provided a better fit to the experimental data. The kinetic data fit the pseudo-second-order model well, with a correlation coefficient of 0.99. The analysis of thermodynamic parameters (ΔG°, ΔH° and ΔS°) revealed that the adsorption process of silver ion by spirulina biomass was exothermic and spontaneous (ΔG° < 0), and exothermic (ΔH° < 0) process. The biosorption capacity of biomass A. platensis serves as a basis for the development of green technology for environmental remediation.     

Antiviral and virucidal activities against herpes simplex viruses of umesu phenolics extracted from Japanese apricot

Umesu phenolics were obtained from the salt extracts of Japanese apricot (Nanko‐mume cultivar of Prunus mume Sieb. et Zucc.) as purified phenolics. The antiviral activities of umesu phenolics obtained were then examined against herpes simplex virus type 1 (HSV‐1) and type 2 (HSV‐2), enveloped DNA viruses. The phenolics inhibited the multiplication of these viruses when added to the culture media of the infected cells. This inhibition occurred at phenolic concentrations at which they showed no severe cytotoxicity. One‐step growth experiments showed that the eclipse period in the HSV‐1 multiplication process was extended in the presence of umesu phenolics and that the addition of phenolics after the completion of viral DNA replication did not affect their multiplication. More drastic effects were observed on virucidal activities against HSV‐1 and HSV‐2; the infectivity decreased to 0.0001 when infected cells were incubated with 3 mg/ml phenolics at 30°C for 5 min. These results demonstrate the antiviral and virucidal activities of umesu phenolics and suggest a potential pharmacological use for these phenolics as a sanitizing or preventive medicine against superficial HSV infections.     

钝顶螺旋藻（Spirulina platensis）藻胆体Langmuir-Blodgett膜

从钝顶螺旋藻中分离制备完整藻胆体 ,然后滴加于空气 水界面上 ,应用LB膜技术制备藻胆体LB膜。结果表明 ,藻胆体在空气 水界面上具有很好的成膜性能。将藻胆体LB单层膜转移到刚揭开的云母表面 ,喷一层金 ,然后用扫描隧道显微镜观察。结果表明 ,藻胆体在Langmuir Blodgett膜中的排列方式与其在体内类囊体膜表面的排列方式类似 ,一排排聚集在一起 ,然后排列成膜。藻胆体的“核”吸附在云母表面 ,而藻胆体的“杆”伸向外面。由于钝顶螺旋藻易于规模化培养 ,藻胆体容易批量制备 ,加之藻胆体具有的独特的光物理、光化学特性和良好的成膜性能 ,以及本身就是纳米量级的颗粒 (5 0 70nm) ,预示着藻胆体在纳米光电子器件中具有很好的应用前景。     

Antiviral activity of Basidiomycete mycelia against influenza type A(serotype H1N1) and herpes simplex virus type 2 in cell culture

In this study, we investigated the in vitro antiviral activity of the mycelia of higher mushrooms against influenza virus type A(serotype H1N1) and herpes simplex virus type 2(HSV-2), strain BH. All 10 investigated mushroom species inhibited the reproduction of influenza virus strain A/FM/1/47(H1N1) in MDCK cells reducing the infectious titer by 2.0–6.0 lg ID50. Four species, Pleurotus ostreatus, Fomes fomentarius, Auriporia aurea, and Trametes versicolor, were also determined to be effective against HSV-2 strain BH in RK-13 cells, with similar levels of inhibition as for influenza. For some of the investigated mushroom species—Pleurotus eryngii, Lyophyllum shimeji, and Flammulina velutipes—this is the first report of an anti-influenza effect. This study also reports the first data on the medicinal properties of A. aurea, including anti-influenza and antiherpetic activities. T. versicolor 353 mycelium was found to have a high therapeutic index(324.67), and may be a promising material for the pharmaceutical industry as an anti-influenza and antiherpetic agent with low toxicity. Mycelia with antiviral activity were obtained in our investigation by bioconversion of agricultural wastes(amaranth flour after CO2 extraction), which would reduce the cost of the final product and solve some ecological problems.     

钝顶螺旋藻FtsZ在大肠杆菌中的表达和定位

FtsZ是与真核微管蛋白类似的原核骨架蛋白,能在细胞分裂位点聚合组装成环状结构而调控细胞分裂过程。为了研究钝顶螺旋藻（Spirulina platensis）FtsZ蛋白的功能,构建了钝顶螺旋藻FtsZ与绿色荧光蛋白GFP融合表达的质粒,并在大肠杆菌中进行了表达和定位研究,结果发现,表达融合蛋白GFP-FtsZ的大肠杆菌细胞由短杆状变为长丝状,且菌丝体长度与融合蛋白的表达量呈正比。在荧光显微镜下观察到融合蛋白GFP-FtsZ在长丝状体细菌中呈有规律的点状分布,这说明FtsZ蛋白功能高度保守,钝顶螺旋藻FtsZ蛋白能识别大肠杆菌分裂位点并装配成环状结构调控大肠杆菌细胞分裂,FtsZ蛋白的过量表达能抑制大肠杆菌正常的细胞分裂而导致长丝状体细胞的形成。     

鄂尔多斯高原碱湖的钝顶螺旋藻光合生理研究

鄂尔多斯高原碱湖的钝顶螺旋藻光合色素含量高低排列为藻胆素>叶绿素a>类胡萝卜素;各色素具有特定的吸收光谱,活体吸收光谱体现出了各色素的吸收;各色素的荧光发射主峰波长约长于活体的13￣35nm,相对荧光强度约是活体的11倍。其光合速率的日变化呈单峰曲线,13:00时达到最高;光补偿点为28￣30μmol.m-2.s-1;光饱和点为220￣235μmol.m-2.s-1;光合作用的最适温度为35℃。呼吸速率日变化随温度的升高呈缓慢上升的趋势。     

Antiviral Activity of Trichothecene Mycotoxins (Deoxynivalenol,Fusarenon-X,and Nivalenol) against Herpes Simplex Virus Types 1 and 2

The effect of trichothecene mycotoxins, deoxynivalenol (DON), fusarenon-X (FX) and nivalenol (NIV), on plaque formation of herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) in HEp-2 cells was examined. The 50% effective concentrations (EC₅₀) of DON, FX, and NIV for HSV-1 plaque formation were 160, 56, and 120 ng/ml, respectively. Those for HSV-2 plaque formation were 94, 26, and 50 ng/ml, respectively. These three mycotoxins showed about 2-fold higher selectivity to HSV-2 than to HSV-1. Plaque formation of HSV-1 was not inhibited with trichothecenes at concentrations completely inhibiting plaque formation when cells were treated during virus adsorption period or 15 hr before infection. These results indicate that trichothecenes affect replication of HSV-1 after virus adsorption, but not before or during virus adsorption to the host cells.     

Antiviral activity of Basidiomycete mycelia against influenza type A (serotype H1N1) and herpes simplex virus type 2 in cell culture

In this study, we investigated the in vitro antiviral activity of the mycelia of higher mushrooms against influenza virus type A(serotype H1N1) and herpes simplex virus type 2(HSV-2), strain BH. All 10 investigated mushroom species inhibited the reproduction of influenza virus strain A/FM/1/47(H1N1) in MDCK cells reducing the infectious titer by 2.0–6.0 lg ID50. Four species, Pleurotus ostreatus, Fomes fomentarius, Auriporia aurea, and Trametes versicolor, were also determined to be effective against HSV-2 strain BH in RK-13 cells, with similar levels of inhibition as for influenza. For some of the investigated mushroom species—Pleurotus eryngii, Lyophyllum shimeji, and Flammulina velutipes—this is the first report of an anti-influenza effect. This study also reports the first data on the medicinal properties of A. aurea, including anti-influenza and antiherpetic activities. T. versicolor 353 mycelium was found to have a high therapeutic index(324.67), and may be a promising material for the pharmaceutical industry as an anti-influenza and antiherpetic agent with low toxicity. Mycelia with antiviral activity were obtained in our investigation by bioconversion of agricultural wastes(amaranth flour after CO2 extraction), which would reduce the cost of the final product and solve some ecological problems.