Analysis of the Therapeutic Effect of Sodium Copper Chlorophyllin Tablet in Treating60Cases of Leukopenia

Objective： To evaluate the efficacy and safety of sodium copper chlorophyllin （trademarked as ＂Yebaike Tablet （叶拜克） which is abbreviated as YBK in treating leukopenia. Methods： One hundred and five patients with leukopenia caused by various factors were randomized into 3 groups. The 60 patients in the YBK group took orally YBK Tablets at the dose of 40 mg, three times per day, the 30 patients in the leucogen group were treated with Leucogen Tablets （利血生片） at the dose of 20 mg, three times per day, and the 15 patients in the placebo group were administered with vitamin C tablets 100 mg, three times per day. All the subjects were treated for 1 month. The change of peripheral leucocytes count after treatment and adverse drug reaction that occurred in patients were studied. Results： In the 60 patients treated with YBK, the treatment proved to be markedly effective in 34 cases, effective in 17 and ineffective in 9, the total effective rate being 85%, which was significantly higher than that in the placebo group （26.7%, P〈0.01） and similar to that in the leucogen group （83.3 %, P〉0.05）. No adverse reaction was found in the treatment course. Conclusion： YBK can be used in the treatment of leukopenia caused by various factors, satisfactory in efficacy and safe in use.

ISOLATION OF HUMAN TRANSCRIPTS EXPRESSED IN 16HBE CELLS RELATED TO CHLOROPHYLLIN ANTITRANSFORMING ACTIVITY AGAINST ANTI-BPDE BY cDNA REPRESENTATIONAL DIFFERENCE ANALYSIS

Objective: To analyze the differentially expressed cDNA sequences related to chlorophyllin (CHL) mediated inhibition of malignant transformation of human bronchia1 epithelial cell line (16HBE). Methods: 16HBE cells treated with chlorophyllin and anti-BPDE were conducted as tester, 16HBE cells treated only with anti-BPDE were conducted as driver, and cDNA representational difference analysis (cDNA RDA) was used to compare the differential gene expression between the two kinds of cells. The cDNA fragments were ligated to pGEM-T vector and transformed into JM109 bacteria. The plasmid DNA was sequenced and compared with database in GenBank by BLASTN. Results: Among the 5 cloned cDNA sequences, three were novel and were registered in dbEST database, two showed sequence homology to alpha-enolase and a newly found gene ribosomal protein S18/S6-like. Conclusion: These 5 cDNA sequences might play important roles in antitransforming effect of chlorophyllin.

Study on the Photodynamic Efficiency of Sodium Chlorophyllin from Spirulina

[Objectives] This study was conducted to investigate the photodynamic technology(PDT) of water-soluble sodium chlorophyllin extract from Spirulina and its photodynamic sterilization efficiency on Gram bacteria and photodynamic antitumor effect on rat glioma C6 cells. [Methods]The absorption spectrum and fluorescence spectrum of sodium chlorophyllin were measured by an ultraviolet spectrophotometer and a fluorescence spectrophotometer;the plate count method was used to investigate the photodynamic sterilization efficiency of sodium chlorophyllin on Escherichia coli and Staphylococcus aureus;and the MTT method was used to determine the photodynamic antitumor effect of sodium chlorophyllin on rat glioma cell C6. [Results] The sterilization rates of sodium chlorophyllin with the 100 J/cm^2 photodynamic treatment were 98.96% 1.284 and 100% 0 on S. aureus and E. coli, respectively. The half maximal inhibitory concentration(IC50) of the sodium chlorophyllin photodynamic therapy with 5, 10 and 20 J/cm^2 on C6 were 69.9, 48.21 and 47.56 μg/ml respectively, and the dark toxicity was extremely low at 0 J/cm^2. [Conclusions]The photodynamic treatment mediated by the alcohol-extracted sodium chlorophyllin from Spirulina showed excellent inhibitory effects on bacteria and tumor cells. This study initially reveals its excellent photodynamic performance and provides a reference for its in-depth application in the field of photodynamic therapy.

Effects of Sodium Copper Chlorophyllin on Mesenchymal Stem Cell Function in Aplastic Anemia Mice

Objective： To investigate the effects of sodium copper chlorophyllin （SCC） on the proliferation, differentiation and immunomodulatory function of mesenchymal stem cells （MSCs） from mice with aplastic anemia. Methods： A mouse model of aplastic anemia was established by exposure of BALB/c mice to sublethal doses of 5.0 Gy Co60 γ radiation, followed by transplantation of 2 × 108 lymph node cells from DBA/2 donor mice within 4 h after radiation. Aplastic anemic BALB/c mice were randomly divided into six groups： the treated groups, which received 25, 50, or 100 mg/kg/day SCC, respectively; a positive control group treated with cyclosporine A （CsA）; and an untreated model control group （model group）; while, the non-irradiated mice as the normal control group. SCC or CsA were administered by gastrogavage for 20 days, starting on day 4 after irradiation. Peripheral blood cells were counted and colony-forming fibroblasts （CFU-F） in the bone marrow were assayed. The ability of MSCs to form calcium nodes after culture in osteoinductive medium was also observed. The immunosuppressive effect of MSCs on T lymphocytes was analyzed by enzyme-linked immunosorbent assay and flow cytometry, to evaluate the efficacy of SCC in mice with aplastic anemia. Results： Peripheral blood white cell and platelet counts were increased by medium and high SCC doses, compared with the untreated control. CFU-Fs were also increased compared with the untreated control, and the numbers of calcium nodes in MSCs in osteoinductive medium were elevated in response to SCC treatment. The percentage of Forkhead box protein 3 （FOXP3^＋） T cells was increased in T celI-MSC cocultures, and the cytokine transforming growth factor 131 was up-regulated in SCC-treated groups. Conclusion： The results of this study suggest that SCC not only promotes the proliferation and differentiation of MSCs, but also improves their immunoregulatory capacity in mice with apiastic anemia.

顶空进样-气相色谱法测定着色剂叶绿素铜钠盐中溶剂残留量

建立顶空进样-气相色谱测定着色剂叶绿素铜钠盐中溶剂残留量的分析方法。对平衡温度、分流比及色谱柱型号等参数进行考察,氢火焰离子化检测器(Flame Ionization Detector,FID)检测,内标法定量。结果表明,通过CD-624毛细管色谱柱分离,平衡温度设为80℃时效果最优,6种残留溶剂可实现完全分离,线性关系良好,r均大于0.999,加标回收率为84.50%~98.83%,相对标准偏差(n=6)均低于10%。该方法简单、快速、准确,可满足着色剂叶绿素铜钠盐中溶剂残留量的检测。

First report on chlorophyllin to protect mammalian and fish muscle cells from pesticide toxicity via activation of p53 and PARP

Objectives:Pesticide toxicity has become one of the major environmental menaces affecting all types of life forms of the ecosystem.Pesticides get washed off from agricultural fields into nearby water bodies and enter the aquatic organisms.Their bio-accumulated form finally reaches the human race,through consumption of pesticide infested aquatic animals,causing several physiological dysfunctions.Hence it becomes necessary to find a therapeutic cure/a preventive measure to stop the health hazard issues of pesticide.With this projection a search for a phyto-based-product was made whose primary objective would be to lower the pesticidal toxicity in fish and simultaneously in the human race.Methods:In this study we tried to check whether the phyto-chemical,Chlorophyllin(CHL),known for its anti-genotoxic,anti-oxidant activities,could render any kind of protection against Cypermethrin(CM)induced-toxicity in fish model and mammalian cell line L6.Both the model L6 and fish were pre-treated with CHL prior to exposure of CM.Different scientific parameters like%cellular cytotoxicity,reactive oxygen species(ROS)generation,nuclear condensation,etc were checked to validate the possibility of CHL in protecting CM-induced toxicity.Results:The overall results revealed that pre-treatment with CHL could restrict the ROS generation leading to modulation in associated cytokine proteins expression NFkβand IFNγ.Further,CHL lowered nuclear condensation and elevated expression of DNA repair proteins p53 and PARP,showing a kind of pre-activation of signalling cascades for overall protection against the severity of pesticidal toxicity.Conclusion:Thus,this phyto-based preventive approach would possibly solve many areas of human health issues related to pesticide toxicity in future.

酪蛋白胶束与多元活性分子的相互作用及其复合物特性

以酪蛋白胶束(casein micelles,MC)为基质,以单一原花青素(proanthocyanidin,PC)、儿茶素(catechin,Cat)、叶绿素铜钠(chlorophyllin sodium copper salt,Chl)或其二元组合为配体,通过热处理或热处理联合超声处理制备二元、三元复合物,系统分析了单一PC、Cat、Chl或其二元组合物与MC的相互作用,解析MC对多元分子的同时结合能力,表征复合物结构、微观形貌及热稳定性,探究三元复合物的抗氧化性以及体外模拟消化特性。结果表明,单一PC、Cat、Chl通过疏水相互作用与MC形成二元复合物,其中MC对Chl表现出较高的亲和力。热处理条件下,当MC以疏水作用结合一个分子后,另一个分子则以范德华力、氢键结合于MC上,且298 K时第1个分子的结合提升了第2个分子与MC的结合常数和结合位点数。PC、Cat、Chl单一分子或其二元组合物的结合对MC空间结构及微观形貌无显著影响。热处理条件下,活性分子的结合增加了MC的热稳定性。就抗氧化性而言,热处理改善了三元复合物的2,2-联氮双(3-乙基苯并噻唑啉-6-磺酸)阳离子自由基清除力,而热处理联合超声处理改善了1,1-二苯基-2-三硝基苯肼自由基清除活性。此外,多元活性分子的同时负载改善了MC在模拟肠液中的消化性。本研究结果可为MC在多元活性分子负载中的应用提供一定参考。

模拟叶片反射的光散射仿生膜光谱分析

为应对高光谱成像侦察,设计了2种模拟植物叶片反射谱的光散射膜并分别命名为TiO_(2)/Chl和TiO_(2)/SCC仿生膜。两类膜的基体均为亲水的聚乙烯醇/氯化锂薄膜,散射微粒均为二氧化钛(TiO_(2)),色素分别为叶绿素(Chl)和叶绿素铜钠盐(SCC)。采用分光光度计分别测量了样品和桂花树叶片的反射率及桂花树叶片色素提取液、Chl和SCC溶液的吸光度。结果表明,在430~2500 nm内,TiO_(2)具有强散射特性,增强了基体膜的反射辐射。在TiO_(2)的散射作用下,由于Chl和SCC具有与植物叶片色素相似的吸收特性,且基体膜含水,仿生膜反射谱均呈现与绿色叶片反射谱相似的4个基本特征,即“绿峰”“红边”“近红外高原”和水吸收带。TiO_(2)/Chl和TiO_(2)/SCC仿生膜的反射谱与桂花树叶片的相关系数分别为0.90和0.95。

生血宁治疗缺铁性贫血的临床研究

蚕砂中提取叶绿素、经皂化、铁代精制成铁叶绿酸钠,并以此为原料制成生血宁片,治疗缺铁性贫血180例,以硫酸亚铁片作对照。结果成人治疗组120例,治愈108例占90％,显效12例占10％;成人对照组30例治愈10例,占33.33％,显效8例占26.67％,有效6例占20％,无效6例占20％。儿童治疗组60例,治愈52例占86.67％,显效8例占13.33％。儿童对照组30例,治愈13例占43.33％,显效5例占16.67％,有效4例占13.33％,无效8例占26.67％,生血宁片疗效明显优于硫酸亚铁片,且无明显毒副反应,易为广大病人接受值得推广应用。

生血宁治疗缺铁性贫血的临床研究

目的观察生血宁治疗缺铁性贫血的药效及对铁代谢平衡调节的分子机制。方法 1 0 0例缺铁性贫血患者随机分为生血宁治疗组和葡萄糖酸亚铁对照组各 5 0例 ,治疗组用生血宁 0 1 g ,每天 3次口服 ;对照组用葡萄糖酸亚铁 0 1 g ,每天 3次口服 ;均连续服用 3 0天。治疗前后各检测 1次血清铁 (Fe)、总铁结合力 (totalironbindingcapacity ,TIBC)、转铁蛋白饱和度 (transferrinsaturation ,TS)、铁蛋白 (serumferritin ,SF)、转铁蛋白 (transferrin ,Tf)、血清可溶性转铁蛋白受体 (solubletransferrinreceptor ,sTfR)、血常规 ,并进行中医气血两虚证候评分。结果治疗组总有效率为 92 %;表明生血宁能改善铁代谢 ,提高Fe、TS、SF ,降低TIBC、Tf、sTfR ;促进红细胞生成作用明显 ;尚能促进白细胞及血小板生成。对照组总有效率为 3 2 %。两组比较差异有显著性 (P <0 0 1 )。结论生血宁治疗缺铁性贫血及气血两虚证效果明显 ,能改善铁代谢 ,提高Fe、TS、SF ,降低TIBC、Tf、sTfR。

由竹叶制取叶绿素铜钠盐的研究

本文研究了以竹叶、杨树叶等为原料制取叶绿素铜钠盐的工艺步骤和工艺条件,结果表明,用95％乙醇为主要溶剂萃取,经过5％NaOH溶液皂化,再用CuSO_4进行铜化反应,并以NaOH成盐等步骤。所得叶绿素铜钠盐产品符合国家食品添加剂标准GB3262-82,得率0.25％～0.3％(以鲜竹叶为基准)。

桑叶叶绿素锌钠盐的制备及其稳定性研究

利用桑叶制备叶绿素锌纳盐溶液,并对其稳定性进行研究。经过研究得到结论:叶绿素锌钠盐水溶性较好,耐热性、耐氧化还原性、pH为4.00～11.00时的稳定性较好,但光照条件下,稳定性较差。常见的食品添加剂(蔗糖、葡萄糖、食盐、柠檬酸)和一些金属离子(Al^(3+)、Ca^(2+)、Na^+、Mg^(2+))对叶绿素锌钠盐的稳定性无影响,而Cu^(2+)、Fe^(2+)对叶绿素锌钠盐的稳定性有较大的影响。

大叶黄杨叶制备叶绿素铜钠盐的工艺研究

以鲜大叶黄杨叶为原料 ,用 95 %酒精为溶剂提取叶绿素 ,再通过化学法制得叶绿素铜钠盐。工艺条件试验证明 :选用 5 % Na OH溶液进行皂化 ,再用 1 0 % Cu SO4 溶液进行铜代 ;最后用丙酮溶解 ,5 % Na OH乙醇溶液成盐 ,制得叶绿素铜钠盐达到国家标准。

海藻酸钠为囊材锐孔法制备铁叶绿酸钠微囊

研究了采用海藻酸钠为囊材,CaCl2溶液为固化液,锐孔法制备铁叶绿酸钠微囊的工艺条件。在实验中,探讨了固化液的浓度、囊材浓度、针头孔径、下滴高度、搅拌速度、乳化剂浓度等实验参数对微囊形态及包封率的影响。结果表明:以海藻酸钠为囊材锐孔法制备铁叶绿酸钠微囊的最佳工艺条件是海藻酸钠的浓度为1%,CaCl2溶液的浓度为2%,海藻酸钠溶液与铁叶绿酸钠溶液的浓度比为1:1,针头孔径0.45mm,转速800r/min,下滴高度10cm。铁叶绿酸钠微囊产品为墨绿色小颗粒,外形颗粒圆整,大小均匀,赋有青草味。

姜叶叶绿素的提取及叶绿素铜钠的制备

研究姜叶叶绿素的提取工艺,并进行叶绿素铜钠的制备及其稳定性的初步分析。结果表明,提取姜叶叶绿素的最佳工艺条件为:以2:1的丙酮-95%乙醇混合溶液作提取溶剂,料液比1:30、浸提温度60℃、浸提时间5h;按先皂化后铜代工艺制备的叶绿素铜钠产品,具一定耐酸性,90℃以下热稳定性较好。

叶绿素铜钠盐的制备

用正交法考察了温度、ＮａＯＨ用量、溶剂Ａ和溶剂Ｂ的体积比，以及溶剂总体积（ｍｌ）和糊状叶绿素重量（ｇ）比等四因素对叶绿素铜酸制备过程中的皂化工序的影响；对铜代过程中硫酸铜用量和铜代条件进行试验；探索了酸化结晶工序的工艺条件；采用了反萃结晶法精制叶绿素铜酸；测定了叶绿素铜酸在溶剂中的溶解度；研究了反萃结晶过程的工艺条件；考察了蒸发干燥过程中温度和时间的影响．在对叶绿素铜钠盐制备过程的总结和分析的基础上，提出了优化的工艺流程和条件，并对其进行了实验验证．全过程叶绿素铜钠盐的收率可达５９．８％．

叶绿素镁钠盐对液态食品中Staphylococcus aureus的光动力杀菌研究

以叶绿素镁钠盐为光敏剂,研究了其对几种液体食品中金黄色葡萄球菌的光动力杀菌效果。结果表明,叶绿素镁钠盐对几种液态食品中的金黄色葡萄球菌具有很强的杀菌效果,其中以10-5mol/L的叶绿素镁钠盐杀菌效果最佳,10 min金黄色葡萄球菌减少了4.5个对数;随着光照时间的延长,液态食品中的金黄色葡萄球菌的致死率明显增加,特别是在起始10 min内;一般透光性好的食品包装材料对液态食品中金黄色葡萄球菌光动力杀菌效率没有影响,但液态食品的pH对其杀菌效率有一定的影响。对于不同的液态食品,在澄清或酸性的食品中叶绿色镁钠盐的光动力杀菌效果较好,但当食品中含固体颗粒或较浑浊时,其杀菌效率显著降低。

新型茶绿色素叶绿素锌钠盐的研究

利用低档茶叶及其废弃物制备新型纯天然水溶性茶绿色素叶绿素锌钠盐,并对其稳定性进行研究。结果表明:该色素水溶性极好,呈鲜艳的绿色,稳定性强,是一种优质的天然营养保健食用色素。

复配叶绿素铜钠对草莓的保鲜研究

探讨了复配叶绿素铜钠保鲜涂膜在常温下对草莓的货架期的影响,通过草莓腐烂度、总酸度、VC和可溶性糖的含量变化来评定保鲜效果。结果表明:用叶绿素铜钠、NOCC和丙二醇-1,2复配制成的保鲜涂膜具有较好的水溶性和成膜性,在贮藏期间能有效降低草莓的呼吸强度,减少水分及营养物质的损失,推迟草莓的衰老,使草莓的货架期达到5d,比未处理的草莓货架期延长4d。

叶绿素铁钠盐的制备

以苦菜中提取的叶绿素为原料,经皂化、酸化、铁代、成盐制备叶绿素铁钠盐,用紫外-分光光度法检测其质量。单因素试验和响应曲面法结合优化工艺试验条件为:先在pH≈12的体系中,59℃下皂化90 min;再于pH≈2～3体系中,65℃下酸化60 min后,加入42 mL质量分数10%FeSO4溶液,铁代时间97 min;最后用10%NaOH-乙醇调节溶液pH≈12而成盐。