蜜环菌菌丝体化学成分的研究——Ⅴ.蜜环菌辛素和蜜环菌壬素的分离与鉴定

从人工液体发酵培养的蜜环菌[Armillaria mellea(Vahl.ex Fr.)Quel.]菌丝体的石油醚提取物中,经硅胶柱层析,低压柱层析和制备性薄层层析分离得到两个新的原伊鲁烷型倍半萜芳香酸酯类化合物。命名为蜜环菌辛素(armillarilin)和蜜环菌壬素(armillarinia),根据光谱(UV,IR,¹HNMR,¹³CNMR和MS)分析,推定它们的化学结构分别为Ⅰ和Ⅱ。     

蜜环菌菌丝体化学成分的研究——Ⅶ.丙酮提取物中化学成分的分离与鉴定

自人工液体发酵生产的白磨科(Tricholomataceae)蜜环菌[Armiilaria meilea(Vahl.exFr.)Quel.]菌丝体的丙酮提取物中分离得到八个化合物,其中三个为原伊鲁烷型倍半萜醇芳香酸酯类化合物。根据光谱(UV,IR,¹H-NMR,¹³C-NMR,2D-NMR和MS)分析,其中一个为已知物4′-methylmelledonal(Ⅰ),两个为新化合物即密环菌丑素(armillaritin)和蜜环菌寅素(armillarivin),其结构分别为式Ⅱ和Ⅲ。另外五个为已知物赤藓醇(D-erythritol),甘露醇(Dmannitol),杜鹃花酸(azclaic acid)、苔藓酸(orsellinic acid)和甘油醇-α-单油酸酯(glycerin-α-monooleate)。     

蜜环菌菌丝体化学成分的研究——Ⅴ.蜜环菌辛素和蜜环菌壬素的分离与鉴定

从人工液体发酵培养的蜜环菌[Armillaria mellea(Vahl.ex Fr.)Quel.]菌丝体的石油醚提取物中,经硅胶柱层析,低压柱层析和制备性薄层层析分离得到两个新的原伊鲁烷型倍半萜芳香酸酯类化合物。命名为蜜环菌辛素(armillarilin)和蜜环菌壬素(armillarinia),根据光谱(UV,IR,~1HNMR,~(13)CNMR和MS)分析,推定它们的化学结构分别为Ⅰ和Ⅱ。     

蜜环菌菌丝体化学成分的研究——Ⅵ.蜜环菌癸素(armillaripin)的分离与结构

从人工发酵培养的蜜环菌[Armillaria mellea(Vahl.ex Fr.)Quel.属白蘑科(Tricholometaceae)真菌]菌丝体的石油醚提取物中,经硅胶层析分离得到一新的倍半萜醇芳香酸酯类化合物,命名为蜜环菌癸素(armillaripin)。根据光谱(UV,IR,~1HNMR,~(13)CNMR和MS)分析推测其化学结构,并进一步用X-线衍射晶体解析法确证其结构和其相对构型。     

蜜环菌酒的研制

目的蜜环菌酒富含多糖,具有天麻酒的相似药理活性,可作为天麻酒的替代品。方法将富含多糖的蜜环菌发酵液进行蜜环菌酒生产工艺的优化,制备蜜环菌酒。结果蜜环菌酒的主要成分为酒精16%、多糖0.301mg/mL、蛋白质44.9mg/L、氨基酸3.30g/L、总酸4.8g/L、总酯3.21g/L。结论蜜环菌酒色、香、味、体纯正,含有功能成分。     

蜜环菌菌丝体化学成分的研究——Ⅶ.丙酮提取物中化学成分的分离与鉴定

自人工液体发酵生产的白磨科(Tricholomataceae)蜜环菌[Armiilaria meilea(Vahl.exFr.)Quel.]菌丝体的丙酮提取物中分离得到八个化合物,其中三个为原伊鲁烷型倍半萜醇芳香酸酯类化合物。根据光谱(UV,IR,~1H-NMR,~(13)C-NMR,2D-NMR和MS)分析,其中一个为已知物4′-methylmelledonal(Ⅰ),两个为新化合物即密环菌丑素(armillaritin)和蜜环菌寅素(armillarivin),其结构分别为式Ⅱ和Ⅲ。另外五个为已知物赤藓醇(D-erythritol),甘露醇(Dmannitol),杜鹃花酸(azclaic acid)、苔藓酸(orsellinic acid)和甘油醇-α-单油酸酯(glycerin-α-monooleate)。     

HPLC法测定蜜环菌中麦角甾醇含量

目的建立蜜环菌中麦角甾醇含量的测定方法。方法采用高效液相色谱方法,采用Diamonsil(TM)C18分析柱(250mm×4.6mm,5μm),流动相为甲醇,流速1.0mL/min,检测波长283nm;柱温25℃。结果麦角甾醇进样量在2～8μg之间与峰面积呈良好的线性关系,r=0.9999;平均回收率为99.72%,RSD为0.19%。方法具有较高的专属性、准确性、重现性和可行性。结论该方法简便、准确、线性范围宽、灵敏度好,为蜜环菌质量评价提供了可靠依据。     

银杏蜜环口服溶液中单糖HPLC特征图谱及其质量评价

目的 利用HPLC建立银杏蜜环口服溶液中糖的特征图谱，并对不同来源的银杏蜜环口服溶液质量进行评价。方法 采用1-苯基-3-甲基-5-吡唑啉酮（1-phenyl-3-methyl-5-pyrazolone，PMP）柱前衍生HPLC，结合中药指纹图谱相似度分析软件对4批次市售银杏蜜环口服溶液和6批次自制口服溶液中单糖进行相似度分析，同时通过对组方中成分单糖组成进行测定，对共有峰进行了归属和确认。结果 通过对自制口服溶液与市售口服溶液的HPLC特征图谱进行研究，确定了5个共有峰，经过归属验证，分别为甘露糖、鼠李糖、葡萄糖、半乳糖和阿拉伯糖，相似度介于0.998~1.000之间；同时对口服溶液及其组方中原药材进行了单糖组成测定，发现水解口服液中单糖来源于蜜环菌粉、银杏叶提取物及矫味剂甜菊糖；进一步对不同厂家的银杏叶提取物和蜜环菌粉进行单糖组成测定，结果显示不同厂家的银杏叶提取物和蜜环菌粉单糖组成有明显差异。最后通过对口服液及其原辅料中多糖进行提取、测定，发现口服液中多糖主要来源于蜜环菌粉。结论 本研究建立的方法能较全面地体现银杏蜜环口服溶液组分中单糖的特征，将市售口服溶液、自制口服溶液及其原药材的水解单糖进行归属及溯源，可用于口服液及其原料药材、辅料的质量评价。     

蜜环菌多糖对小鼠免疫功能的影响

蜜环菌多糖(100mg/kg·d×5d,ig)能显著增加正常小鼠和注射环磷酰胺所致免疫功能抑制小鼠的溶血素生成。当剂量为50mg/kg·d×5d,ig时,它还能显著增加正常小鼠的空斑形成细胞数。体外试验,蜜环菌多糖(10、50μg/ml)显著增强刀豆素A诱导的小鼠淋巴细胞增殖反应,但对二硝基氯苯所致小鼠迟发型过敏反应无显著增强作用。此外,蜜环菌多糖还能增加小鼠静注碳粒廓清速率及腹腔巨噬细胞的吞噬百分数和吞噬指数。     

复方天麻蜜环菌片治疗紧张性头痛临床观察

目的：观察复方天麻蜜环菌片治疗紧张性头痛的疗效及安全性。方法：100例紧张性头痛患者随机分为两组各50例。治疗组予复方天麻蜜环菌片250 mg,po tid;对照组给予吲哚美辛肠溶片25 mg,po tid。疗程均为28 d。结果：治疗后两组患者头痛发作次数均较治疗前减少（P〈0.05）,疼痛持续时间缩短（P〈0.05）,疼痛积分和生活质量指数明显改善（P〈0.001）;两组间治疗后比较,头痛发作次数和持续时间、生活质量积分差值、疼痛缓解和生活质量改善的总有效率差异均有统计学意义（P〈0.05）。治疗组1例患者因不良反应而退出试验。两组不良反应发生率分别为20.0%和28.0%。结论：复方天麻蜜环菌片治疗紧张性头痛疗效显著,安全性高,与传统标准治疗相比某些指标尚有优势。     

榆耳发酵液中新倍半萜——榆耳三醇的结构

从榆耳(Gloeostereum incarnatum S.Ito et Imai)的液体发酵物中经层析分离得一新倍半萜榆耳三醇(gloeosteretriol),根据红外光谱、核磁共振谱和质谱数据推定其结构为Ⅰ,并用X-射线晶体衍射进一步确证其结构和立体构型。     

Identification and characterization of nitric oxide synthase inSalmonella typhimurium

The presence of the nitric oxide synthase (NOS) enzyme from Salmonella typhimurium (S. typhimurium) was identified by measuring radiolabeled L-[3H]citrulline and NO, and Western blot analysis. NOS was partially purified by both Mono Q ion exchange and Superose 12HR size exclusion column chromatography, sequentially. The molecular weight of NOS was estimated to be 93.3 kDa by Western blot analysis. The enzyme showed a significant dependency on the typical NOS cofactors; an apparent Km for L-arginine of 34.7 mM and maximum activity between 37 degrees C and 43 degrees C. The activity was inhibited by NOS inhibitors such as aminoguanidine and N(G),N(G)-dimethyl-L-arginine. Taken together, partially purified NOS in S. typhimurium is assumed to be a different isoform of mammalian NOSs.     

温敏性吲哚美辛/β-环糊精包合物的制备及体外评价

目的合成兼具温敏性及药物包合能力的新型药物载体聚(N-异丙基丙烯酰胺)-β-环糊精(PNIPA-β-CD)，以吲哚美辛为模型药物，考察该载体的释药行为。方法末端带羧基的PNIPA与改性后的环糊精衍生物在1-(3-二甲氨基丙基)-3-乙基-碳二亚胺(EDC)的作用下缩合，得到PNIPA-β-CD，采用冻干法制备吲哚美辛/PNIPA-β-CD包合物。红外、¹H NMR和DSC表征载体的结构及包合物的形成；用分光光度法测定载体材料的LCST，并进行包合物体外释药研究。结果PNIPA-β-CD在35 ℃发生相转变，吲哚美辛/PNIPA-β-CD包合物的载药量为5.8％，药物与载体摩尔比为0.97∶1。体外释放研究表明吲哚美辛/PNIPA-β-CD包合物在37 ℃的释药比其在25 ℃的释放要慢，在LCST以上具有一定的缓释作用。结论该载体既具有温敏性，又具有药物包合作用，并且在体温条件下具有缓释作用，是一种新型的温敏性药物载体。     

异叶梁王茶甙Ⅲ和Ⅳ的结构

继前报后,又从异叶梁王茶[Nothopanax davidii(France)Harms]树皮中分离得到两种五环三萜皂甙。经化学和波谱(IR,¹HNMR,~(13)CHMR,¹H-¹HCOSY,¹³C-¹H COSY,MS)分析,分别鉴定为3-O-α-(2’,4’-O-二乙酰基)-L-吡喃阿拉伯糖-3β-羟基齐墩果-12-烯-28,29-双羧酸-28-O-[α-L-吡喃鼠李糖(1-4)-β-D-吡喃葡萄糖(1—6)-β-D-吡喃葡萄糖]酯甙(Ⅲ),命名为异叶梁王茶甙Ⅲ和3-O-β-(2-O-乙酰基)-D-吡喃木糖-3β-羟齐墩果-12-烯-28,29-双羧酸甙(Ⅳ),命名为异叶梁王茶甙Ⅳ。这两种皂甙均系首次从植物中分出的新化合物。     

Comparative analysis of methods to measure aerosols generated by a vibrating mesh nebulizer.

Different approaches have been employed for in vitro assessment of the aerosol particle size generated by inhalation devices. In this study, aerosols from the Omron MicroAir vibrating mesh (VM) nebulizer were measured by cascade impaction (CI) using the MSP Next Generation Pharmaceutical Impactor (NGI), the ThermoAndersen Cascade Impactor (ACI), and by time-of-flight (TOF) analysis with the TSI 3321 Aerodynamic Particle Sizer Spectrometer (APS). The VM nebulizer was evaluated with sodium fluoride (NaF; 2.5%) and with generic albuterol (0.083%). Aerosol particle size (MMAD), respirable fractions (RF < 5 microm), and fine particle fractions (FPF < 3.3 microm) were determined with each method at room temperature (RT) and 4 degrees C using 50% average relative humidity. By NGI at either RT or 4 degrees C, aerosol particle sizes were similar for both NaF and albuterol (4.3-4.5 microm MMAD) with 55-61% RF and 27-43% FPF. With ACI, the distribution of particles at RT was similar except at the extremes of the dispersion and the MMAD was smaller (3.3 microm MMAD; p = 0.03). At 4 degrees C, particle sizes determined by ACI results were similar to the NGI (MMAD 4.1 microm; p > 0.05). TOF analysis by APS with albuterol gave significantly larger calculated MMAD (cMMAD) than either CI method (7.2 microm; p < 0.001). TOF measurements of nebulized albuterol at RT and 4 degrees C were equivalent. In summary, the results of VM nebulized NaF and albuterol were more consistent and generally equivalent when determined by NGI (at RT and 4 degrees C) and ACI analysis (at 4 degrees C). In contrast, aerosol particle sizes measured by TOF in the APS at both RT and 4 degrees C were larger than results obtained by CI. Differences in aerosol particle distribution obtained by different analysis methods should be considered while evaluating the in vitro performance of VM nebulizers.     

Characterization and Modulation of the Transferrin Receptor on Brain Capillary Endothelial Cells

PURPOSE: The expression level of the transferrin receptor (TfR) on brain capillary endothelial cells (BCECs) and the endocytosis of 125I-transferrin (125I-Tf) by this receptor was investigated. Furthermore, the influence of iron, the iron scavenger deferoxamine mesylate (DFO), astrocytic factors, a GTP-ase inhibitor (tyrphostin-A8, T8), lipopolysaccharide (LPS), and the radical scavenger N-acetyl-L-cysteine (NAC) on the TfR expression was studied to gain insight in the use and optimization of the TfR for drug targeting to the brain. METHODS: Experiments were performed with primary cultured bovine BCECs that were incubated with 125I-Tf at 4 degrees C (to determine binding) or at 37 degrees C (to determine endocytosis) in the absence or presence of the modulators. For full saturation curves in the absence or presence of iron or DFO, analysis was performed with a population approach using NONMEM, allowing us to estimate a single value for affinity (Kd, concentration of 50% receptor occupancy) and separate values for maximum receptor occupancy (B(max). RESULTS: On BCECs, the TfR is expressed extracellularly (B(max) of 0.13 fmol/microg cell protein), but also has a large intracellular pool (total B(max) of 1.37 fmol/microg cell protein), and is actively endocytosing Tf via clathrin-coated vesicles. At 4 degrees C, a Kd of 2.38 microg/ml was found, whereas the Kd at 37 degrees C was 5.03 microg/ml. Furthermore, DFO is able to increase both the extracellular as well as the total binding capacity to 0.63 and 3.67 fmol/microg cell protein, respectively, whereas it had no influence on Kd. B(max) at 37 degrees C after DFO preincubation was also increased from 0.90 to 2.31 fmol/microg cell protein. Other modulators had no significant influence on the TfR expression levels, though LPS increased cellular protein concentrations after 2-h preincubation. CONCLUSIONS: The TfR is expressed on BCECs and actively endocytoses Tf, making it a suitable target for drug delivery to the bloodbrain barrier and the CNS. DFO up-regulates the TfR expression level, which may influence targeting efficiency.     

黄褐毛忍冬化学成分的研究

从我省商品药材金银花的主要品种之一,黄褐毛忍冬(Lonicera fulvotomentosa Hsu et S.C.Cheng)的花蕾中分离到五种以常春藤皂甙元构成甙元的皂甙(Ⅰ～Ⅴ),本文报道化合物Ⅰ,Ⅱ和Ⅴ的结构测定。Ⅴ为新的三萜酸双糖链甙,命名为黄褐毛忍冬皂甙甲(fulvotomentoside A),Ⅰ和Ⅱ为已知化合物,分别被鉴定为α-常春藤皂甙(α-hederin)和无患子皂甙B(sapindoside B)。     

Characteristics of chitosanases from<Emphasis Type="Italic">Aspergillus fumigatus</Emphasis> KB-1

Two chitosanases produced by Aspergillus fumigatus KB-1 were purified by ion exchange and size exclusion chromatographies. Molecular weights of chitosanases were 111.23 kDa (chitosanase I) and 23.38 kDa (chitosanase II). The N-terminal amino acid sequence of chitosanase II was determined as follows: YNLPNNLKQIYDKHKGKXSXVLAKGFTN. The optimum pH of the chitosanase I and II was 6.5 and 5.5, respectively. The optimum temperatures were 60 degrees C for chitosanase land 70 degrees C for chitosanase II. Hydrolysis products of two chitosanases were analyzed by HPLC and GPC. Chitosanase I hydrolyzed substrate to glucosamine. Chitosanase II produced chitooligosaccharides.     

Effect of storage on the plasma concentration of chlorpromazine and six of its metabolites.

The concentrations of chlorpromazine (CPZ) and six of its metabolites in patient plasma samples that had been stored for 24 h at -20 degrees C, for 1 week at -20 degrees C, and for 4 weeks at -70 degrees C were compared. The concentrations of CPZ and six of its metabolites in human plasma spiked with known concentrations and stored at -70 degrees C for either 3 or 12 months were also compared. No significant difference was seen in the concentrations after storage under nitrogen and analysis by a high-performance liquid chromatography technique.     

Ambient temperature effects on 3,4-methylenedioxymethamphetamine-induced thermodysregulation and pharmacokinetics in male monkeys.

Changes in ambient temperature are known to alter both the hyperthermic and the serotonergic consequences of 3,4-methylenedioxymethamphetamine (MDMA). Metabolism of MDMA has been suggested to be a requisite for these neurotoxic effects, whereas the hyperthermic response is an important contributing variable. The aim of the present study was to investigate the interaction between ambient temperature, MDMA-induced thermodysregulation, and its metabolic disposition in monkeys. MDMA (1.5 mg/kg i.v.) was administered noncontingently at cool (18 degrees C; n = 5), room (24 degrees C; n = 7), and warm (31 degrees C; n = 7) ambient temperatures. For 240 min following MDMA administration, core temperature was recorded and blood samples were collected for analysis of MDMA and its metabolites 3,4-dihydroxymethamphetamine (HHMA), 3,4-dihydroxyamphetamine, and 3,4-methylenedioxyamphetamine (MDA). A dose of 1.5 mg/kg MDMA induced a hypothermic response at 18 degrees C, a hyperthermic response at 31 degrees C, and did not significantly change core temperature at 24 degrees C. Regardless of ambient temperature, plasma MDMA concentrations reached maximum within 5 min, and HHMA was a major metabolite. Curiously, the approximate elimination half-life (t(1/2)) of MDMA at 18 degrees C (136 min) and 31 degrees C (144 min) was increased compared with 24 degrees C (90 min) and is most likely because of volume of distribution changes induced by core temperature alterations. At 18 degrees C, there was a significantly higher MDA area under the concentration-time curve (AUC) and a trend for a lower HHMA AUC compared with 24 degrees C and 31 degrees C, suggesting that MDMA disposition was altered. Overall, induction of hypothermia in a cool environment by MDMA may alter its disposition. These results could have implications for MDMA-induced serotonergic consequences.