蚕沙叶绿素提取及合成脱镁叶绿酸工艺改进研究

目的探讨不同提取工艺对蚕沙中提取叶绿素及制取叶绿素衍生物脱镁叶绿酸的影响,以提高蚕沙叶绿素及叶绿素衍生物的制备量。方法单因素实验设计,以蚕沙软化时间(含水量)、不同提取溶剂为考察条件,优化蚕沙叶绿素提取工艺,并探讨浓盐酸脱镁时间对叶绿素合成脱镁叶绿酸制备率的影响。结果蚕沙软化时间2 h(含水量26%)、丙酮∶乙醇体积比1∶1做溶剂为提取叶绿素的最佳条件,提取率提高到1.43%。以蚕沙叶绿素粗品合成叶绿素衍生物脱镁叶绿酸,浓盐酸脱镁搅拌反应72 h制备率达670 mg.g 1。结论通过改良工艺,提高了蚕沙叶绿素及其衍生物脱镁叶绿酸的提取制备率。     

脱镁叶绿环类衍生物的分离,制备及鉴定

自蚕砂糊状叶绿素分得的叶绿素 a(1),于不同条件下经酸降解、脱羧焦化、甲酯化、氧化,在各步中可得脱镁叶绿素 a(2)、脱镁叶绿酸 a(3),焦脱镁叶绿酸 a(4)、甲基脱镁叶绿酸 a(5)、甲基焦脱镁叶绿酸a(6)、10-羟基甲基脱镁叶绿酸 a(7)和红紫素7-内酯二甲酯(8)。7和8的 ~1HNMR 和 MS 数据为首次报道。     

脱镁叶绿酸的制备及其光敏抗菌活性

以初步纯化的叶绿素为原料,利用叶绿素酶水解、选择性酸降解两种方法制备脱镁叶绿酸。高效液相色谱分析显示两种方法得到的产物图谱相似,酸降解产物中脱镁叶绿酸A的含量较高。抗菌实验表明脱镁叶绿酸对革兰阳性菌具有光敏抑菌效果,对革兰阴性菌和酵母无明显作用。     

蚕沙粗品叶绿素酸降解产物的分离与鉴定

从蚕沙粗品叶绿素酸降解产物中分离制得在肿瘤中有选择性潴留作用,光动力学活性强于癌光啉的脱镁叶绿甲酯一酸 a 类组分混合物。经 HPLC 制备分离,~1H-NMR 和 MS 证明,它们为脱镁叶绿甲酯一酸 a(58%),脱镁叶绿甲酯一酸 a′(11%)和焦脱镁叶绿酸 a(31%)三种成分。     

从蚕砂中制备脱镁叶绿酸的工艺研究

目的 :优化以蚕砂为原料制备脱镁叶绿酸工艺条件 ,提高产出率。方法 :以叶绿素提取率为指标 ,对脱镁叶绿酸制备的乙醇法和丙酮法工艺进行比较 ,用正交试验法对丙酮法的工艺条件进行优化。结果 :脱镁叶绿酸的最佳制备条件为蚕砂粉碎度60目 ,加1/10倍药材量的水软化5h以上 ,加3倍药材量的溶媒 ,提取温度70℃ ,回流时间60min ,提取次数2次 ,酸化时 pH调至2 5。结论 :软化水用量、丙酮量对叶绿素提取率影响较大 ,经优化后可提高丙酮法制备脱镁叶绿酸的产出率4 6倍     

亚硒酸锰（Ⅱ）—叶绿酸钠的制备及其对小鼠的急性毒性和小鼠肝癌H …

目的:制备亚硒酸锰(Ⅲ)-叶绿酸钠,并测定其对小鼠的急性毒性及对小鼠肝癌H22的抑制情况.方法:以蚕沙提取叶绿素,由叶绿素制备脱镁叶绿酸,与锰络合后制得锰(Ⅲ)叶绿酸,再以亚硒酸化合制得亚硒酸锰(Ⅲ)-叶绿酸钠;测定亚硒酸锰(Ⅲ)-叶绿酸钠的急性毒性及对小鼠肝癌H22的抑制情况.结果:作者首次合成了亚硒酸锰(Ⅲ)-叶绿酸钠.测得其LD50=1175 mg.kg-1(95%的可信区间为851～1621 mg.kg-1;其对小鼠肝癌H22的抑癌率为45～47%(灌胃剂量为500 mg.kg-1).结论:亚硒酸锰(Ⅲ)-叶绿酸钠常温下性质稳定,毒性小,对小鼠肝癌H22有明显的抑制作用,可进一步做临床试验.     

亚硒酸锰(Ⅲ)-叶绿酸钠的制备及其对小鼠的急性毒性和小鼠肝癌H22的初步试验

目的:制备亚硒酸锰(Ⅲ)-叶绿酸钠,并测定其对小鼠的急性毒性及对小鼠肝癌H22的抑制情况.方法:以蚕沙提取叶绿素,由叶绿素制备脱镁叶绿酸,与锰络合后制得锰(Ⅲ)叶绿酸,再以亚硒酸化合制得亚硒酸锰(Ⅲ)-叶绿酸钠;测定亚硒酸锰(Ⅲ)-叶绿酸钠的急性毒性及对小鼠肝癌H22的抑制情况.结果:作者首次合成了亚硒酸锰(Ⅲ)-叶绿酸钠.测得其LD50=1175 mg.kg-1(95%的可信区间为851～1621 mg.kg-1;其对小鼠肝癌H22的抑癌率为45～47%(灌胃剂量为500 mg.kg-1).结论:亚硒酸锰(Ⅲ)-叶绿酸钠常温下性质稳定,毒性小,对小鼠肝癌H22有明显的抑制作用,可进一步做临床试验.     

二氢卟吩e6衍生物的简便制备

蚕沙叶绿素粗提物经酸降解得到脱镁叶绿酸a粗品，不经分离，直接与亲核试剂如甲醇、乙胺发生开环反应，一步得到二氢卟吩e6衍生物。     

脱镁叶绿一酸a荧光特性研究

目的:研究中药蚕砂中脱镁叶绿一酸a的荧光性质。方法:采用丙酮提取,Al2O3柱层析和乙醚萃取自蚕砂中分离脱镁叶绿一酸a,采用荧光扫描确定其激发波长和发射波长,考察pH值、温度及光照时间对其荧光强度的影响。结果:脱镁叶绿一酸a的激发波长和发射波长分别为665和675 nm。pH碱性、提高温度可降低其荧光强度,光照则可提高其荧光强度。结论:脱镁叶绿一酸a在中性、酸性和低温条件下稳定,光照可加强其荧光效应。     

铬叶绿酸钠的合成及临床试用初探

采用脱镁叶绿酸与铬络合后制成铬叶绿酸钠口服降糖药,并对１１例糖尿病患者在使用降糖药同时加用铬叶绿酸钠口服进行初步临床验证,观察患者血糖、血胆固醇（ＴＣ）、血甘油三酯（ＴＧ）水平的变化。结果证明铬叶绿酸钠降低血糖、血脂有一定作用。     

氢氧化钙糊剂与樟脑酚液治疗乳牙根尖周炎的效果观察

目的观察对比氢氧化钙糊剂与樟脑酚液用于乳牙根尖周炎消毒的疗效比较。方法对268例乳牙根尖周炎进行根管治疗前的消毒,随机平均分成两组,分别封氢氧化钙糊剂与樟脑酚液棉捻一周,对比消毒效果。结果氢氧化钙糊剂组成功129例,成功率96.27%,樟脑酚液组成功102例,成功率76.12%。两者存在显著性差异（P〈0.01）。结论氢氧化钙糊剂是乳牙根尖周炎根管消毒的安全有效的理想药物,值得临床推广使用。     

Characterization of two isomeric beta-d-glucosiduronic acids derived from 1,2-diethyl-3-hydroxypyridin-4-one (CP94) in rat liver homogenate incubates

1,2-Diethyl-3-hydroxypyridin-4-one (CP94) is an orally active iron chelator with potential for use in photodynamictherapy. This investigation reports the formation and characterization of two isomeric glucuronides of CP94 in rat liver homogenate incubates. To assign the glucuronidation sites in the CP94 molecule, two O-methylated derivatives of CP94 have been synthesized. By comparing the spectral characteristics of the CP94 3-O- and 4-O-methyl derivatives with CP94 and the CP94 glucuronides formed during incubation, evidence was obtained which enabled the assignment of these two isomeric glucuronides to the 3-O-glucuronide and 4-O-glucuronide of CP94. It was found that the 3-O-glucuronide was the dominant CP94 metabolite under in-vitro conditions. In an attempt to understand the potential influence of structural variation on the glucuronidation of CP94 analogues, the 1-and 2-monoethyl derivatives of CP94 were investigated. The 2-monoethyl derivative of CP94 yielded only the 3-O-glucuronide in rat liver homogenate incubate, while no glucuronide was formed from the 1-monoethyl derivative. In addition, no glucuronide from the 3-O-methyl or 4-O-methyl derivatives of CP94 could be detected. The relevance of these findings to the development of new 3-hydroxypyridin-4-one iron chelators is discussed.     

脑瘫患儿血钙、磷、碱性磷酸酶和骨矿物质含量的研究

目的：探讨脑瘫患儿血Ca P碱性磷酸酶（ALP）及骨矿物质含量的变化。方法 40名脑瘫患儿和30名正常对照用全自动血液分析仪测血清Ca P ALP用单光子BMD400E型骨密度仪测量桡骨远端1／3处骨矿物质含量（BMC／BW）。结果：脑瘫患儿组血清Ca P与对照组无差异（P＞0．05），ALP则明显高于对照组（P＜0．01），桡骨远端1／3处骨矿物质含量（BMC／BW）低于对照组（P＜0．05）。结论：脑瘫患儿血清Ca P通过机体自身的调节可基本维持正常；而ALP升高和骨密度减低则提示脑瘫患瘫患儿骨矿化明显低于正常对照组。     

不同根管治疗方法引起根管治疗期间痛的差异性分析研究

目的通过对比传统逐步后退法与改良逐步后退法根管预备后采用氢氧化钙与甲醛甲酚（FC）根管封药后对根管治疗期间痛（EIP）的控制及影响情况,评价不同根管预备方法和根管封药对于EIP的影响。方法 120例需要进行根管治疗的病人,随机分入A、B、C、D四组。A、C组均使用逐步后退法预备根管后,A组封FC,C组封氢氧化钙。B、D组均使用改良逐步法预备根管后,B组封FC,D组封氢氧化钙糊剂。根管充填时评价约诊间急症情况。结果在使用相同的根管药物情况下,不同的根管预备方法对于EIP的影响没有显著性差异。但是对于相同的根管预备方法,不同的根管封药对EIP影响有显著性差异（P〈0.05）。结论氢氧化钙比甲醛甲酚能更有效降低根管治疗期间疼痛和肿胀的程度。     

Enhanced clearance of Escherichia coli and Staphylococcus aureus in mice treated with cyclophosphamide and lactoferrin

Previous studies on cyclophosphamide (CP)-immunocompromised mice showed accelerated reconstitution of immune system function following oral treatment with lactoferrin (LF). The aim of this investigation was to evaluate the ability of mice, treated with a sublethal dose of CP and given LF, to combat bacterial infections. Mice were injected with a single, intraperitoneal dose of CP (350 mg/kg body weight). One group of CP-treated mice was also given LF in drinking water (0.5% solution) for 14 days. Untreated and LF-treated mice served as controls. On day 15 following CP administration, mice were infected intravenously with 10(8) Escherichia coli or 5 x 10(7) Staphylococcus aureus. Twenty-four hours later, the number of colony-forming units (CFU) in spleens and livers were determined. Phenotypic analysis of blood leukocytes was determined, as well as the ability of splenic and peritoneal cells to produce IL-6 spontaneously and in the presence of lipopolysaccharide (LPS). Treatment with CP, or with CP and LF, led to profound reduction of E. coli CFU in the liver and the spleen; treatment with LF alone had significant inhibitory effects on organ enumerated CFU. S. aureus CFUs were also significantly reduced in spleens of mice treated with CP or CP/LF and, to a lesser degree, after LF alone. These effects were also significantly reduced in the livers. Analysis of blood cellular phenotype revealed total number of peripheral leukocytes was lower in the CP-treated group (52.6%) but not significantly different from control values in CP/LF and LF-treated groups (90.7% and 104.6%, respectively). Conversely, percentage of blood neutrophils was markedly elevated in CP and CP/LF groups--62% and 42.5% vs. 18.4% in controls. These findings were accompanied by production of IL-6 by splenic and peritoneal cells which was significantly increased in CP- and CP/LF-treated groups. It was concluded that the increased clearance of bacteria in the organs of mice treated with CP and CP/LF may result from a rise in the number of neutrophils infiltrating the organs and contributing to accelerated clearance of bacteria. The study also suggests that the ability of cells from CP- and CP/LF-treated mice to produce significantly more IL-6 may also contribute to increased resistance to infections. Lastly, together with our previous data, this study indicates that LF used to reconstitute the antigen-specific immune response in CP-treated mice does not impair their resistance to infection.     

Rats bred for differences in preference to cocaine: other behavioral measurements.

Cocaine has repeatedly been shown to produce conditioned place preference (CPP) in the rat. The present study employed the heterogenous N/Nih rat stock to produce a selectively bred rat line determined by individual place preference to a conditioning dose of 2.5 mg/kg cocaine. As each of three generations of rats were exposed to the CPP task, cocaine-preferring (CP) males were mated with CP females whereas cocaine-nonpreferring (CNP) male rats were paired with their female counterparts. Rats in litters of the third generation of these selectively bred rats were used in two collateral studies: one involving the discriminative stimulus properties of cocaine and the other to investigate the ability of cocaine to stimulate activity. Results indicate that the continued breeding of CP animals has resulted in rats that prefer cocaine, whereas the breeding of CNP rats is defining a line of rats that actually find cocaine aversive. In testing the discriminative stimulus performance of five male CP and five male CNP rats, the learning rates and dose-response relationship to cocaine were not significantly different between these two groups. In contrast, administration of 5.0 and 7.5 mg/kg cocaine to male and female CP and CNP rats indicated that, although all groups were stimulated by cocaine when compared to vehicle administration, male CNP rats showed a significantly decreased reaction to these two doses of cocaine. The possibility that conditioned place preference and locomotor stimulation are subserved by the same neural substrates, that is, most probably the dopaminergic systems in the nucleus accumbens of the brain, is discussed.     

Non-cell Corynebacterium parvum generated by nanotechnology: a promising immunomodulator with less side effects

Corynebacterium parvum (CP), a kind of immunomodulator, has been well documented in immunotherapy to tumor. However, severe side effects, such as intrahepatic granulomas and scleromas in injected areas, restrict its clinical application. To minimize side effects of CP, a non-cell Corynebacterium parvum product (NCPP) was prepared by disposing CP with Nanotechnology. In present study, we compared effect of NCPP with that of CP and found: (1) NCPP with non-formaldehyde residue was easy to be absorbed without swelling and scleroma in local injected areas; (2) NCPP caused no obvious liver injury in murine and macaques; (3) NCPP maintained powerful anti-tumor activity, increased splenic index, elevated macrophage number, phagocytosis and production of hydrogen peroxide (H(2)O(2)), and nitric oxide (NO); (4) Importantly, unparallel CP, NCPP could stimulate macrophages to produce low level of tumor necrosis factor-alpha(TNF-alpha) but high level of interferon-gamma (IFN-gamma), an inhibitor to fibrosis. Our study has led to the view that NCPP will evolve into a new valuable immunomodulator for clinical application.     

Permeation of 4-cyanophenol and methyl paraben from powder and saturated aqueous solution through silicone rubber membranes and human skin

The objectives were to compare permeation from neat powder and saturated aqueous solution of two model compounds into homogeneous silicone rubber (polydimethylsiloxane) membranes (SRM) and human skin, which is heterogeneous, and to test the common assumption that solid chemicals do not absorb unless liquid is present. The steady-state flux of 4-cyanophenol (CP) through SRM from the powder (0.0684 +/- 0.0040 mg/cm2 x h) was almost the same as from a saturated solution (0.0789 +/- 0.0064 mg/cm2 x h, indicating that solid chemicals can absorb without the presence of liquids. The steady-state flux of CP through skin of a single subject was much smaller from the powder (0.0118 +/- 0.0064 mg/cm2 x h) than from the saturated solution (0.168 +/- 0.033 mg/cm2 x h). The average flux for powder relative to the saturated aqueous solution was 7.24% in skin compared with 87.2% in SRM for CP and 9.02% in skin compared with 99.9% in SRM for methyl paraben. It is evident that absorption into SRM and skin can occur from powdered chemicals and that surface oils or moisture are unnecessary. However, SRM proved to be a poor surrogate for dermal permeation from powders of CP and MP.     

Metabolic activation and cytotoxicity of cyclophosphamide in primary cultures of postnatal rat hepatocytes

We have developed a model of primary cultures of postnatal rat hepatocytes to characterize the metabolic activation of xenobiotics to toxic intermediates and to study the mechanism(s) by which these chemicals produce cellular injury. This model was employed to investigate the cytochrome P-450 mediated biotransformation of cyclophosphamide (CP) to cytotoxic metabolites that nonspecifically alkylate DNA and cellular proteins. The parenchymal cells were isolated by an in situ collagenase perfusion technique and cultured for 24 hr prior to drug treatment. The cultures were then exposed to CP concentrations ranging from 1 × 10^?4 M to 1 × 10^?3 M for 24 hr. Initial studies indicated minimal toxicity to non-replicating parenchymal hepatocytes maintained in ornithine-supplemented, arginine-deficient medium. The addition of arginine permitted the overgrowth of fibroblasts in the same culture system. These fibroblasts then became the target of alkylating CP metabolites produced by the par-enchymal cells. By day 3 after CP administration, cell number and total protein per dish decreased by over 40 percent. The morphology of the cultures changed dramatically because of fibroblast destruction. The cytotoxicity to dividing fibroblasts was eliminated by administering 2-diethylaminoethyl-2, 2-diphenylvalerate hydrochloride (SKF 525-A), an inhibitor of the cytochrome P-450 monooxygenase system, to the co-cultures treated with CP. The alkylating metabolites of CP produced by the parenchymal cells and released into the culture medium were quantitated by reacting aliquots of medium from CP-treated cells with 4-(p-nitrobenzyl)pyridine. These results provide both direct and indirect evidence of drug metabolism in cultured cells and suggest that this co-culture system can be utilized to evaluate the metabolic activation of xenobiotics.     

The estrogenic proliferative effects of two alkylphenols and a preliminary mechanism exploration in MCF‐7 breast cancer cells

Bisphenol A (BPA) and 4‐cumylphenol (4‐CP), as estrogen‐like chemicals, are ubiquitous in the environment media and associated with the occurrence and development of hormone‐dependent tumors. However, the combinatorial effects of these two structurally similar alkylphenols are not well informed. In the present study, the classic breast cancer cell line MCF‐7 was used as in vitro model to estimate the estrogenic proliferative effects of BPA and 4‐CP. MTT assay, reactive oxygen species, cell apoptosis, cell cycle, and real‐time fluorescent quantitative Step One Plus Real‐time PCR System (Applied Biosystems, CA, USA) were applied to explore their proliferative mechanisms. MTT results showed that both BPA and 4‐CP ranging from 10^?9 to 10^?5 M stimulated cell proliferation in a nonmonotonic dose‐response manner. Along with the proliferative effects, cell cycle was progressed from G0/G1 to S and G2/M phase. Meanwhile, the expression levels of ERα, pS2, and Bcl‐2 mRNA were also upregulated. In contrast, 4‐CP and BPA at high dose (10^?4 M) obviously displayed antiproliferative effects in MCF‐7 cells via inducing cell apoptosis and blocking cell cycle in G0/G1 phase. As expected, the relative expression levels of ERα, pS2, and Bcl‐2 mRNA were decreased, whereas Bax mRNA was increased. Interestingly, the proliferative or antiproliferative effects of 4‐CP were higher than that of BPA. Moreover, coexposure of lower concentrations BPA and 4‐CP significantly induced cell proliferation in a synergistic manner. These findings indicated that the potential environmental risks of coexposure of BPA and 4‐CP were greater than either of them.