白花前胡丙素的结构修饰研究

目的对白花前胡丙素［(+)-praeruptorin A］进行结构修饰,寻找活性化合物。方法首先从白花前胡(Peucedanum praeruptorum)根中分离得到白花前胡丙素,从白花前胡丙素出发,用碱水解及各种酰化反应,半合成各种结构修饰产物。结果得到18个(2～19)凯林内酯类化合物,通过IR,¹HNMR和MS等方法确定其结构。结论其中14个(5～18)为新化合物,这些新化合物均有不同程度的钙离子拮抗活性,但作用强度均较白花前胡丙素弱。     

缺氧条件下白花丹醌对肝癌HepG2细胞增殖、凋亡与侵袭及HIF-1α表达的影响

目的 研究白花丹醌在缺氧条件下对人肝癌细胞HepG2增殖、凋亡、侵袭和低氧诱导因子1α(hypoxia induced factor1α，HIF-1α)及其下游基因表达的影响。方法 采用二氯化钴(CoCl₂)化学诱导法建立HepG2细胞体外缺氧模型，经不同浓度(2，4，8 μmol·L^–1)白花丹醌处理24 h后，MTT、平板克隆形成试验观察HepG2细胞增殖水平变化；使用Annexin V/PI双染流式细胞术检测HepG2细胞凋亡情况；使用Transwell试验观察HepG2细胞侵袭能力的变化；使用qRT-PCR检测HepG2细胞内HIF-1A mRNA转录水平变化；使用Western blotting检测细胞内HIF-1α及其下游基因c-Myc、VEGFA、MMP9和TWIST1的蛋白表达水平。结果 当CoCl₂处理浓度为150 μmol·L^–1时，HepG2细胞增殖水平未见显著变化，而HIF-1α蛋白表达水平显著升高(P<0.01)，提示体外缺氧模型建立成功。与常氧对照组相比，MTT、平板克隆形成试验结果显示不同浓度白花丹醌作用HepG2细胞后，其增殖水平受到显著抑制(P<0.05或P<0.01)； Transwell试验结果显示白花丹醌可显著抑制HepG2细胞侵袭(P<0.05或P<0.01)；流式细胞术结果表明白花丹醌能显著诱导HepG2细胞凋亡(P<0.05或P<0.01)；Western blotting及qRT-PCR检测结果显示，白花丹醌能显著下调HIF-1α蛋白及HIF-1A mRNA表达水平(P<0.05或P<0.01)。Western blotting检测结果显示HIF-1α及其下游基因c-Myc、VEGFA、MMP9和TWIST1的蛋白表达水平均显著下调(P<0.05或P<0.01)。结论 CoCl₂体外模拟肝癌HepG2细胞缺氧的适宜浓度为150 μmol·L^–1；在缺氧条件下，白花丹醌可显著抑制肝癌HepG2细胞的增殖与侵袭，同时诱导细胞凋亡，其作用机制可能与下调HIF-1α蛋白及其下游靶基因蛋白表达有关。     

白花前胡中前胡香豆素D和前胡香豆素E的分离和鉴定

从中药白花前胡(Peucedanum Praeruptorum Dunn)根中分到5个化合物,经理化常数、波谱数据及化学反应分别鉴定为前胡香豆素D(I),Pd-Ib(II),前胡香豆素E(III),nodakenetin(IV)和scopoletin(V)。其中化合物I和III为两个新化合物,与巳知化合物凯林内酯的化学沟通确定了其绝对构型,其化学结构分别为3'(S),4'(S)-二乙酰氧基-3',4'-二氢邪蒿内酯(I)和3'(R)-惕各酰氧基-4'-酮基-3',4'-二氢邪蒿内酯(III)。化合物IV和V为首次从该植物中分离得到。通过DEPT,¹H-¹HCOSY和¹³C-¹HCOSY等实验归属了II的碳氢信号。     

白花前胡中白花前胡甙和Pd-C-I的分离和鉴定

从白花前胡(Peucedanum,praeruptorum)根中分得7个化合物,经化学方法和光谱分析分别鉴定为Pd-C-I(I),白花前胡甙(II),香草酸(III),没食子酸(IV),nodakenin(V),rutarin(VI)和isorutarin(VII)。II为新化合物,其化学结构为4-O-β-D-吡喃葡萄糖基-3-甲氧基苯丙酮,命名为白花前胡甙。I为首次从白花前胡中分得的线型二氢吡喃香豆素类化合物,这对前胡属植物化学分类学有一定意义。还利用2DNMR纠正了文献中关于化合物I和VII的个别碳信号归属的错误。     

白花前胡丙素抗哮喘模型小鼠Th17/Treg失衡作用

目的 研究白花前胡丙素对哮喘模型小鼠体内Th17/Treg失衡的干预作用。方法 小鼠腹腔注射和雾化吸入卵清蛋白以建立气道炎性反应模型,并灌胃不同剂量的白花前胡丙素干预。模型结束后,动物肺功能仪分析气道高反应性;细胞计数器和Diff-Quick染色计数支气管灌洗液中白细胞总数及分类;酶联免疫吸附法检测血清或BALF中细胞因子和炎性介质的水平;苏木精-伊红(HE)染色法观察气道病理改变;流式细胞检测脾脏CD4+细胞数及分类;RT-PCR分析脾脏Foxp3 mRNA的表达。结果 与对照组比较,模型组呈现明显的气道炎性反应和气道高反应(P<0.05或P<0.01);IgE、IL-17水平增加(P<0.01),IL-10、TGF-β1水平降低(P<0.01);CD4⁺CD25⁺Foxp3⁺细胞占总CD4⁺细胞的比例降低(P<0.01);Foxp3 mRNA的表达减少(P<0.01)。与模型组比较,白花前胡丙素组气道炎症显著减轻,中、高剂量组气道反应在氯乙酰胆碱剂量>10 mg·kg^-1均显著受到抑制(P<0.05或P<0.01);白花前胡丙素组IgE水平显著降低(P<0.01),IL-10、TGF-β1水平显著增加(P<0.05或P<0.01),中、高剂量组IL-17显著降低(P<0.01);白花前胡丙素组显著增加CD4⁺CD25⁺Foxp3⁺细胞的比例(P<0.05或P<0.01),并增加Foxp3 mRNA的表达(P<0.05或P<0.01)。结论 白花前胡丙素具有抗哮喘鼠Th17/Treg失衡的作用。     

白花曼陀罗悬浮培养细胞转化对羟基苯甲醛生成天麻素

目的研究天麻素在植物细胞的转化合成。方法利用白花曼陀罗细胞悬浮培养转化外源对羟基苯甲醛合成天麻素，并应用多种色谱技术进行分离纯化，根据转化产物的理化性质和光谱数据分析鉴定结构。结果利用白花曼陀罗细胞成功地将对羟基苯甲醛转化为天麻素(II)，并得到由对羟基苯甲醛生成天麻素的转化中间体对羟基苯甲醇(I)。另外，从细胞培养物中分离得到β-D-吡喃阿洛酮糖(III)和5-正丁氧基-β-D-吡喃阿洛酮糖(IV)两种化合物。结论白花曼陀罗细胞悬浮培养能转化对羟基苯甲醛合成天麻素。     

白花前胡丙素C-3′和C-4′反式结构类似物的半合成白花前胡丙素C-3′和C-4′反式结构类似物的半合成

目的对白花前胡丙素［(+)-praeruptorin A］进行结构修饰,半合成C-3′和C-4′反式结构类似物,寻找活性化合物。方法首先从白花前胡(Peucedanum praeruptorum)根中分离得到白花前胡丙素,从白花前胡丙素出发,运用碱水解及各种酰化反应,半合成各种结构修饰产物。结果首次合成了17个白花前胡丙素C-3′和C-4′反式结构类似物,通过IR,¹HNMR,MS等方法确定它们的结构。结论17个化合物均为新化合物,其中一些新化合物有显著的钙离子拮抗活性,首次证明C-3′和C-4′反式结构的这类化合物同样具有活性。     

前胡香豆素A的分离和结构鉴定

从中药白花前胡(Peucedanum praeruptorum Dunn)根中分离得到7个化合物,经理化常数和波谱数据的测定以及化学反应,鉴定为补骨脂素(Ⅱ)、5-甲氧基补骨脂素(Ⅲ)、8-甲氧基补骨脂素(Ⅳ)、北美芹素(Ⅴ)、peucedanocoumarin Ⅱ(Ⅵ)、β-谷甾醇(Ⅶ)和前胡香豆素A(Ⅷ)。Ⅷ为一新化合物,通过和凯林内酯的化学沟通确定了其绝对构型,化学结构为3′(R)-羟基-4′(E)-惕各酰氧基-3′,4′-二氢邪蒿内酯。Ⅱ～Ⅳ和Ⅶ为首次从白花前胡中分离得到。     

中药白花前胡根中香豆素白花前胡素(E)的分离鉴定

从白花前胡(Peucedanum praeruptorum Dunn)根的醚提取物中除分得已知香豆素白花前胡丙素和另一种结晶外,还分得一种具有生理活性的香豆素单体,命名为(+)白花前胡素(E)[(+)praeruptorin E]。经元素分析、紫外、红外、质谱、核磁、气相及水解产物的鉴定,证明白花前胡素(E)为(+)3′-当归酰基-4′-异戊酰基-顺式-凯林酮(+)3′-angeloyl-4′-isovalerylcis-khellactone,此香豆素对小白鼠具有耐缺氧作用。关于其他成分,将另有报告。     

前胡香豆素B和前胡香豆素C的分离和鉴定

从中药白花前胡(Peucedanum praeruptorum)根中分离和鉴定了5个化合物,经理化常数测定,波谱数据解析及化学反应,分别鉴定为3′-angeloyloxykhellactone(Ⅸ)、前胡香豆素B(Ⅹ)、前胡香豆素C(Ⅺ)、Pd-Ⅲ(ⅩⅢ)和peucedanocoumarin Ⅲ(ⅩⅤ)。其中化合物Ⅹ和Ⅺ为两个新化合物,通过和凯林内酯的化学沟通确定了其绝对构型。其化学结构分别为3′(S)-乙酰氧基-4′(S)-羟基-3′,4′-二氢邪蒿内酯(Ⅹ)和3′(S)-羟基-4′(S)-乙酰氧基-3′,4′-二氢邪蒿内酯(Ⅺ)。Ⅸ为首次报道自天然物中分得。     

刺梨酸的分离与结构研究

从刺梨中分离出一种新的五环三萜酸,命名为刺梨酸。通过光谱分析和衍生物的制备,确定其化学结构为2β,3α,7β,19α-四羟基乌苏-12-烯-28-羧酸。     

陵水暗罗活性成分研究

从陵水暗罗(polyalthia nemoralis A.etDC)根的乙醇提取物中分离得到五个化合物。经光谱(UV,IR,¹H-NMR,¹³C-NMR,DEPT和MS)解析和化学反应,分别鉴定为暗罗素(zincpolyanemine,PN1),2-巯基吡啶-N-氧化物铜盐(PN₂),β-谷甾醇(PN₃),β-谷甾醇-β-D-吡喃葡萄糖甙(PN₄)和2-巯基吡啶-N-氧化物-2-S-β-D-吡喃葡萄糖甙(PN₅),其中PN₅为新化合物,PN2为新天然产物。PN1和PN2有较强抗疟、防霉杀菌作用。PN5的抗疟、防霉试验尚在进行中。     

制川乌配方颗粒与标准煎剂对二甲苯致小鼠耳肿胀抑制作用的比较研究

目的 对制川乌配方颗粒与标准煎剂抗炎药效等效性进行研究。方法 应用二甲苯致小鼠耳肿胀法比较制川乌标准煎剂及配方颗粒对其抑制作用。结果 在抗炎药效等效性研究中,在小鼠耳廓肿胀抑制率22.51%～38.65%共同效应范围内,1/2、1、2倍临床等效剂量（195.00、390.00、780.00 mg/kg）制川乌标准煎剂等效于0.07、0.20、0.46倍临床等效剂量（27.36、77.67、178.29 mg/kg）制川乌配方颗粒所产生的抗炎效应,制川乌配方颗粒等效剂量DEE_{制川乌配方颗粒}=-22.950＋0.258× DEE_{制川乌标准煎剂}（R²=0.958,r=0.979,P<0.01）,回归系数b₀=-22.950,95% CI为（4.549,41.351）,与总体均值0相比,差异有显著性（P<0.05）,常数项b₁=0.258,95% CI为（-0.218,0.299）,与总体均值1相比,差异有显著性（P<0.05）。制川乌配方颗粒与制川乌标准煎剂药剂量在1/8～1倍临床等效应剂量范围内（48.75～390.00 mg/kg）具有可比性,制川乌配方颗粒的等剂量效应EED_{制川乌配方颗粒}=2.963＋1.573×EED_{制川乌标准煎剂}（R²=0.923,r=0.961,P<0.01）,回归系数b₀=2.963,95% CI为（-5.373,11.300）,与总体0比较差异有显著性（P<0.05）,常数项b₁=1.573,95% CI为（1.230,1.915）,与总体均值1相比差异有显著性（P<0.05）。结论 在抗炎方面制川乌配方颗粒和制川乌标准煎剂的等效应剂量不具有等效性,制川乌配方颗粒等效应剂量较制川乌标准煎剂小。在抗炎方面制川乌配方颗粒和制川乌标准煎剂的等剂量效应不具有等效性,制川乌配方颗粒等剂量效应较制川乌标准煎剂大。     

Use of in Vitro Critical Inhibitory Concentration, a Novel Approach to Predict in Vivo Synergistic Bactericidal Effect of Combined Amikacin and Piperacillin Against Pseudomonas aeruginosa in a Systemic Rat Infection Model

Purpose This study was undertaken to explore the use of in vitro critical inhibitory concentration (CIC) as a surrogate marker relating the pharmacokinetic (PK) parameters to in vivo bactericidal synergistic effect [pharmacodynamic (PD)] of amikacin + piperacillin combination against Pseudomonas aeruginosa in a systemic rat infection model. Methods The in vitro antibacterial activities of amikacin and piperacillin, alone and in combinations at various ratios of the concentrations, were tested against a standard [5 × 10⁵ colony-forming units (CFU)/ml] and a large (1.5 × 10⁸ CFU/ml) inoculum of P. aeruginosa ATCC 9027 using a modified survival-time method. The CIC of each individual antibiotic for the different combinations was determined using a cup-plate method. In vivo studies were performed on Sprague-Dawley rats using a systemic model of infection with P. aeruginosa ATCC 9027. PK profiles and in vivo killing effects of the combination at different dosing ratios were studied. Results An inoculum effect was observed with the antibiotics studied. Synergy was seen against both the inocula at the following concentration ratios: 70% C_ami + 30% C_pip and 75% C_ami + 25% C_pip, where C_ami and C_pip are the concentrations of amikacin and piperacillin to produce a 1000-fold decrease in bacterial population over 5 h, respectively. The CIC values determined corroborated with the order of in vitro bacterial killing observed for the antibiotic combinations. The dosing ratio of 12.6 mg/kg amikacin + 36 mg/kg piperacillin (a 70:30 ratio of the individual doses) exhibited the greatest killing in vivo when compared to the other ratios. The PK–PD relationships were described by simple, linear regression equations using the area under the in vivo killing curve as a PD marker and the AUCIC_ami/CIC_ami + AUCIC_pip/CIC_pip, AUC_ami/CIC_ami + AUC_pip/CIC_pip, C_max,ami/CIC_ami + C_max,pip/CIC_pip, and AUCIC_ami/MIC_ami + AUCIC_pip/MIC_pip as PK markers for the amikacin + piperacillin combination. Conclusion The combination of amikacin and piperacillin exhibited synergistic killing effect on P. aeruginosa that could be modeled using CIC as a surrogate marker relating the PK parameters to in vivo bactericidal effect.     

7α和7β-甲基-10β,17β-二乙酰氧基-4-雌甾烯-3-酮的合成

由于7α-甲基或10β-乙酰氧基4(5)烯-3-酮雌(雄)甾化合物具有显著的抗着床或抗蜕膜活性,我们合成了既具有7α-甲基或7β-甲基又具有10β-乙酰氧基的两个新甾族化合物(1_a)和(1_b)。经药理试验表明(1_a)和(1_b)对孕鼠均有抗早孕作用。     

Inhibitory Effect of Fluoranthene on Photosynthetic Processes in Lichens Detected by Chlorophyll Fluorescence

The effect of increasing concentration (0.01, 1 and 5 mg l⁻¹) of fluoranthene (FLT) and the duration of exposure (12, 24, 36, 48 and 60 h) on the chlorophyll fluorescence parameters (F ₀, F _V/F _M, Φ_II and (F _M⁻ F _S)/F _S) of symbiotic algae in thalli of two foliose lichens Lasallia pustulata and Umbilicaria hirsuta was investigated. Also the FLT concentration in thalli of both lichen species and the proportion of symbiotic algae (photobionts) in thalli was studied. The results obtained demonstrated that the applied concentration of FLT (1 and 5 mg l⁻¹) affected primary photochemical processes of photosynthesis of algae in both lichen species. The F ₀ values increased and the F _V/F _M and Φ_II values decreased. The fluoranthene content in thallus of both lichen species was increased with increasing FLT concentration in the environment. Chlorophyll fluorescence parameters showed different sensitivity to the content of FLT in Lasallia pustulata and Umbilicaria hirsuta, respectively. Higher sensitivity, detected in U. hirsuta, can be related to its diverse anatomical structure (higher portion of symbiotic algae in thalli) and to the different (nearly 2.5 times higher) content of FLT after exposure.     

明目蒺藜丸联合聚乙烯醇滴眼液治疗干眼症的临床研究

目的 制备他达拉非片并考察其体内外释药特性。方法 以溶出度为评价指标,筛选他达拉非片各辅料用量及包衣增重。用相似因子（f₂）法比较自制制剂与参比制剂在0.5% SDS溶液、含0.5% SDS的0.1 mol/L的盐酸溶液、含0.5% SDS的pH 4.5醋酸钠缓冲液、含0.5% SDS的pH 6.8磷酸盐缓冲液中溶出曲线的相似性。比较二者在Beagle犬体内的药动学特征。结果 他达拉非片处方为他达拉非20 mg、乳糖50M 227.625 mg、羟丙基纤维素L 10.5 mg、交联羧甲基纤维素钠19.6 mg、SDS 0.525 mg、微晶纤维素M102 70 mg、硬脂酸镁1.75 mg,包衣增质量范围2%～4%。自制与参比制剂在4种溶出介质中的f₂均大于65,二者体外溶出行为相似。2种制剂在Beagle体内的药动学参数AUC_0~t、C_max、t_max均无显著性差异,自制他达拉非片的相对生物利用度为（101.67±8.99）%。结论 成功制备他达拉非片,其体外溶出和体内药动学行为与参比制剂相似。     

五味子乙素抑制M146L细胞Aβ42生成的机制研究

目的研究五味子乙素抑制M146L细胞Aβ₄₂生成的机制。方法体外培养高效表达Aβ42的M146L细胞株，分别加入不同浓度的五味子乙素(1.67，5.00和15.00 μg·mL^-1)、 β分泌酶抑制剂(S4562，100.00 μg·mL^-1)和γ分泌酶抑制剂(S2188，13.68 μg·mL^-1)。用CCK-8(cell counting kit-8)比色法检测不同处理对M146L细胞活性的影响；用ELISA法测定M146L细胞所分泌的Aβ₄₂的变化；用Western blotting检测APP的β分泌酶剪切产物C99蛋白的含量变化，结合用β和γ分泌酶活性试剂盒，检测五味子乙素对这两种酶活性的影响。结果不同处理因素对M146L细胞的存活率均无影响，不具有细胞毒作用。中、高剂量的五味子乙素均不同程度地抑制M146L细胞分泌Aβ₄₂及γ分泌酶的活性，但都不改变M146L细胞C99蛋白的含量及β分泌酶的活性。结论五味子乙素可抑制γ分泌酶活性，其降低Aβ₄₂生成是通过抑制γ分泌酶的活性来实现的。     

The efficacy of Prrrikweg? gel in the treatment of insect bites: a double-blind, placebo-controlled clinical trial

Objective: This study was conducted to examine the efficacy of Prrrikweg^® gel, a homeopathic after-bite gel, in relleving the effects of mosquito bites, in particular itching and erythema. Design: A double-blind, randomized, placebo-controlled clinical trial. Setting: London School of Hygiene and Tropical Medicine. Subjects: 100 healthy volunteers. Methods: All subjects were bitten under laboratory conditions by Aedes aegypti mosquitoes at one spot on the ventral aspect of the left forearm and another on a corresponding position on the right forearm. One spot was treated with the homeopathic after-bite gel and the other with a placebo gel. Main outcome measures: Itching was assessed on a 5-point discrete rating scale at 0, 0.5, 1, 26.5, and 48 h post-bite to compare the itch-relieving efficacy of the two treatments. Erythema development was assessed by photographing the bite sites, measuring length and width of the erythema with at baseline (T₀) to the mean erythema surface at 0.5, 1, 26.5, and 48 h post-bite (T_mean) for the two treatments. Results: Testing erythema development by comparing the ratio T₀/T_{mean, after-bite gel} and the ratio T₀/T_{mean, placebo gel} gave a two-tailed p=0.098 (95% Cl,–0.031–0.361) in favour of the after-bite gel. There was not a statistically significant difference between the itch relief provided by the two treatments (two-tailed p=0.424; 95% Cl,–0.541–0.191). The correlation between itching and erythema was significant (r=0.46; p<0.001). Conclusions: There are strong indications that the homeopathic after-bite gel reduces erythema development following mosquito bites. The homeopathic mother tinctures of Echinacea angustifolia DC., Ledum palustre L, Urtica urens L. as well as the Hamamelis extract in this gel, whether alone or in combination, are the biologically active ingredients. The homeopathic after-bite gel was not demonstrated to relieve itching, however, based on the correlation between erythema and itching, an effect on itching is not inconceivable.     

款冬花多糖抗氧化能力测定

目的用流动注射化学发光法探讨中药款冬花多糖的体外抗氧化作用。方法将款冬花多糖提取液加入3种化学发光体系,测量其发光强度,根据系统化学发光被抑制的程度评价款冬花对活性氧自由基的清除能力,并以抗坏血酸（Vc）为阳性对照。结果款冬花多糖对.OH有很好的清除作用,对O2.、H2O2也有一定的清除作用,且在0.25～4.4 mg.L 1内清除作用随浓度的增加而增强;对自由基的清除能力大小：.OH〉H2O2〉O2.。结论在一定浓度范围内,款冬花多糖具有一定的抗氧化活性。