New procedure for direct bioautographic TLC assay as applied to a tincture of Ranunculus bulbosus

A new procedure for TLC bioautographic assay using a flooding method to inoculate the developed plates is described. Details about several microbiological parameters allow good reproducibility for this method which appears rapid, easy to perform and requiring no specialized equipment. Significantly results obtained with a commercial tincture of Ranunculus bulbosus illustrate this method which can be extended to complex mixtures of natural products in order to isolate active fractions.     

猫爪草多糖RTG-Ⅲ的分离纯化及其生物活性

 目的 研究中药猫爪草水提取液中分离纯化得到一种多糖蛋白复合物——RTG-Ⅲ的生化特性、免疫和抑瘤作用。方法 采用DEAE-Cellulose-52和Superdex 200柱色谱得到均一的多糖组分,电泳和凝胶色谱等方法检测纯度,薄层色谱和气相色谱分析单糖组成,红外光谱确定糖苷键类型,噻唑蓝试验测定免疫活性。结果 RTG-Ⅲ为均一组分,单糖组成为鼠李糖、阿拉伯糖、葡萄糖、半乳糖、甘露糖,其摩尔比为1.00:1.17:51.71:2.16:0.46,总糖含量为80.1％,蛋白含量为12.6％,由谷氨酰胺、谷氨酸等16种氨基酸组成,糖肽键为非O-型。结论 RTG-Ⅲ是以β-D-葡萄吡喃糖为主的一种新的多糖-蛋白复合物,相对分子质量为2.3×10⁴,能够增强免疫细胞对肿瘤细胞HL-60的抑制作用。     

Wound healing and anti-inflammatory properties of Ranunculus pedatus and Ranunculus constantinapolitanus: a comparative study

Ethnopharmacological relevance

In Turkish folk medicine Ranunculus species are used for wound healing and for the treatment of rheumatism. The present study was conducted to evaluate in vivo wound healing and anti-inflammatory properties of Ranunculus pedatus and Ranunculus constantinapolitanus.

Material and methods

In vivo wound healing activity of the extracts prepared from Ranunculus pedatus and Ranunculus constantinapolitanus was evaluated by linear incision and circular excision wound models. Hydroxyproline content of the treated tissues was also assessed. We also studied the anti-inflammatory activity using Whittle method with some modifications.

Results

Methanolic extract of Ranunculus pedatus showed significant wound healing effect both in incision (31.4%) and excision (55.74%) wound models. Methanolic extract of both Ranunculus pedatus and Ranunculus constantinapolitanus demonstrated anti-inflammatory activity with the inhibition value of 26.2% and 23.3% respectively, at the dose of 100 mg/kg. Hydroxyproline content of the tissues treated with the methanolic and aqueous extracts of Ranunculus pedatus and methanolic extract of Ranunculus constantinapolitanus were found to be significantly higher than that of the other extracts.

Conclusion

The experimental data revealed that Ranunculus pedatus showed significant wound healing and anti-inflammatory effect.     

Antimicrobial compounds isolated from Haematoxylon brasiletto

ETHNOPHARMACOLOGICAL RELEVANCE: The decoction of the bark of Haematoxylon brasiletto Karst. (Leguminosae), commonly known as "Palo Brasil", is used in the Mexican traditional medicine to treat mouth and kidney infections, hypertension, stomach upsets, gastric ulcers and diabetes. AIM OF THE STUDY: The present study was performed to evaluate the antimicrobial effects of the methanolic extract of the bark of Haematoxylon brasiletto. MATERIALS AND METHODS: A panel of 12 bacteria and the yeast Candida albicans were used. Minimum inhibitory concentrations (MICs) were determined by the standard broth microdilution method. RESULTS: The results indicate that the extract of Haematoxylon brasiletto inhibited the growth of eight of the tested microorganisms at a concentration limit of 128mug/mL. For the tested compounds the MIC values ranged from 8.7 to 128mug/mL. CONCLUSIONS: The overall results provide promising baseline information for the potential use of the extracts from Haematoxylon brasiletto as well as some of the isolated compounds in the treatment of bacterial infections.     

《中草药》杂志被确认为允许刊载处方药广告的第一批医药专业媒体

目的 研究毛茛Ranunculus japonicus提取物的化学成分.方法 采用硅胶和Sephadex LH-20柱色谱进行成分分离,通过光谱分析鉴定所分离成分的结构.结果 分离获得一个油状样品.该样品经正、反相薄层色谱均为单一斑点,经GC-MS及反相HPLC柱色谱分析均为单一峰.但ESI-MS和1D、2D NMR谱学分析确定其为毛茛苷元(ranunculinin,Ⅰ)、异毛茛苷元(isoranunculinin,Ⅱ)和二氢毛茛苷元(dihydroranunculinin,Ⅲ)3个组分以2∶1∶1比率的混合物.结论 所有化合物均为从该植物中首次分离得到,其中化合物Ⅱ和Ⅲ为首次从天然中分离得到.     

高效液相色谱-脉冲安培电化学检测器法替代微生物检定法测定硫酸新霉素效价的研究

目的建立用于测定硫酸新霉素效价的高效液相色谱-脉冲安培电化学检测器法(HPLC-PAD)。方法建立并验证HPLC-PAD法,测定硫酸新霉素各主要组分含量及相关物质;采用在线膜抑制脱盐技术辅助LC-IT-TOF法测定硫酸新霉素杂质谱,结合组分活性研究确定了硫酸新霉素的主要活性组分;采用半制备液相色谱-蒸发光散射检测器串联制备硫酸新霉素B及硫酸新霉素C精制品,采用波谱手段进行结构确证;采用质量平衡法测定二者的纯度;采用三剂量抗生素微生物检定法测定B、C精制品各自的效价值,推导公式计算出硫酸新霉素的量效转换系数,设计实验进行交互作用考察及实际样品的验证。结果建立的HPLC-PAD方法在分离能力及方法的稳定性上均优于《欧洲药典》(EP)方法,适用于硫酸新霉素各组分的准确定量及有关物质检查;在线膜抑制脱盐技术成功地去除了流动相中的三氟乙酸,对硫酸新霉素的杂质谱进行研究,结合活性研究结果,确定新霉素B和新霉素C为新霉素的主要活性组分;成功制备获得新霉素B、C精制品,通过实验和计算获得了硫酸新霉素的量效转换系数,并通过了验证。结论初步实现了仪器法对微生物生物检定法的替代,基本完成了多组分氨基糖苷类抗生素硫酸新霉素的量效一致性研究工作。     

猫爪草多糖及蛋白含量测定

目的测定猫爪草中多糖及蛋白的含量。方法使用o-T试剂法测定猫爪草中多糖的含量,UV法测定了蛋白含量。结果葡萄糖在0.2~1.0 mg范围内线性关系良好,回归方程为Y=0.782X 0.004 43(R=0.997,n=9)牛血清蛋白在0.05~1.5 mg/m l范围内线性良好,其标准曲线y=0.296 4x 0.027(R=0.998,n=6)。结论结果准确,重现性好,o-T试剂法测多糖含量,不仅可以测总糖的含量,还同时可以测定还原糖的含量,较其他方法操作简单,不必沉淀蛋白质,不受还原性物质干扰。而UV法测定蛋白含量简单,灵敏,快速,不消耗样品,低浓度盐类不干扰测定。多糖与蛋白含量可以同时得出而互相不影响。     

Antibacterial activity of flavanone isolated from Sophora exigua against methicillin-resistant Staphylococcus aureus and its combination with antibiotics

5,7,2′,4′-Tetrahydroxy-8-lavandulyl-flavanone, isolated from Sophora exigua, completely inhibited the growth of 21 strains of methicillin-resistant Staphylococcus aureus (MRSA) at concentrations of 3.13–6.25 μg/mL. The anti-MRSA effect was based on bactericidal action. In combinations of the flavanone with vancomycin, minocycline and rifampicin, the fractional inhibitory concentration indices were 0.90, 0.88 and 0.85, respectively, indicating partial synergistic effects with anti-MRSA antibiotics. The proposed flavanone would be a potent phytotherapeutic agent against MRSA infections.     

Antioxidant properties of scopoletin isolated from Sinomonium acutum

Scopoletin was isolated from Sinomonium acutum and studied using four experimental models designed to assess antioxidant properties. The results indicated that scopoletin scavenged superoxide anion in the xanthine/xanthine oxidase reaction system in a concentration-dependent manner, but did not inhibit xanthine oxidase. Scopoletin may therefore be responsible for the superoxide anion scavenging activity seen in Sinomonium acutum extracts and may be of use in preventing superoxide anion-induced damage in vivo.     

Antiproliferative properties of polyketides isolated from Virola sebifera leaves

An activity-guided fractionation of Virola sebifera Aubl. methylene chloride-soluble fraction provided novel 3,5-dihydro-2-(1'-oxo-3'-hexadecenyl)-2-cyclohexen-1-one (3), two known lignans (1, 2) and dehydro hexadecanoyl resorcinol (4). Isolation and purification were conducted with the application of column chromatography and structures were assigned by spetral analysis (1D and 2D NMR, HREIMS). Compounds 1-4 were evaluated for cytotoxic activities against human tumour cell lines UACC62 (melanoma), MCF-7 (breast), NCI 460 (lung, non-small cells), OVCAR03 (ovarian), PC-03 (prostate), HT-29 (colon), 786-0 (renal) and NCI-ADR (breast expressing phenotype multiple drugs resistance) in vitro. The new polyketide (3) showed selectivity against human OVCAR03 and NCI-ADR cell lines, ranging from 2 to 4 microg/mL.     

毛茛中白头翁素的制备工艺研究

目的:研究白头翁素的制备工艺,以适应工业大生产的需要.方法:单因素考察NaCl浓度、萃取次数对原白翁素粗品收率及干燥温度对白头翁素纯度的影响,并根据理化性质和多种波谱分析(UV,IR,1HNMR,13CNMR,1H-1H COSY,HMQC,HMBC)对其进行结构鉴定及利用高效液相色谱进行纯度测定.结果:最佳制备工艺为NaCl浓度为15%,萃取次数为4次,60℃减压(0.08 Mpa)干燥.白头翁素纯度达99%,收率达0.72‰,质量可控.结论:本制备工艺较为简便,对白头翁素大工业生产提供参考.     

微生物检验在感染性疾病患者预防和诊断治疗中的价值

目的 探讨在感染性疾病患者预防和诊断治疗中应用微生物检验法的价值.方法 对实验中心2019年4月-2020年4月进行检验的80例感染性疾病患者资料进行回顾性分析调查.对照组40例没有经过微生物检验,根据医师临床经验结合患者病情直接给药治疗,观察组则先对患者进行微生物检验,针对患者的检验结果再给予相应的抗生素进行治疗.观...     

猫爪草的无性繁殖

本文报道了在猫爪草的宿根长出幼苗时,把它挖起,然后掰开新苗和单个块根分别进行无性繁殖的方法。此法可较高地提高繁殖指数。     

Anti-cancer properties of terpenoids isolated from Rhizoma Curcumae - A review

Ethnopharmacological relevance

Rhizoma Curcumae is a popular type of traditional Chinese medicine whose essential oils are widely used in the treatment of cancer in China. This review aims to systematically summarize and analyze the anti-cancer properties of terpenoids, the main components of essential oils in Rhizoma Curcumae, and thus enable the development of new anti-cancer drugs.

Materials and methods

Information on the recent progress of anti-cancer studies on terpenoids isolated from Rhizoma Curcumae, including β-elemene, δ-elemene, furanodiene, furanodienone, curcumol, and germacrone, was gathered and analyzed.

Results

Among these terpenoids, β-elemene is the most widely studied, whereas δ-elemene, furanodiene, furanodienone, curcumol, and germacrone have just recently attracted the attention of researchers. The anti-cancer effects of these terpenoids are related to the retardation of cell cycle arrest, the induction of apoptosis, and the inhibition of metastasis or tissue invasion, among others.

Conclusions

Most studies have focused on the in vitro data, and in vivo data is urgently needed. Further insight into the anti-cancer activity and the molecular basis of these compounds, combined with efforts in pharmaceutical chemistry and/or pharmaceutics, will potentially enable the development of new anti-cancer agents.     

云南民间习用水杨梅─—茴茴蒜的生药学研究

茴茴蒜为云南部分地区水杨梅的习用品种，来源于毛茛科毛茛属植物茴茴蒜Ｒａｎｕｎｃｕｌｕｓｃｈｉｎｅｎｓｉｓ的全草。本文对其原植物、性状、显微及理化特征作了初步研究。     

猫爪草化学成分的研究Ⅰ

 目的研究猫爪草的化学成分。方法 经柱色谱分离,并通过理化常数和光谱分析鉴定化合物结构。结果 分离得到6个化合物,分别为棕榈酸乙酯(Ⅰ),肉豆蔻酸(Ⅱ),棕榈酸(Ⅲ),β-谷甾醇(Ⅳ),豆甾醇(Ⅴ),菜油甾醇(Ⅵ)。结论 Ⅰ,Ⅱ,Ⅲ,Ⅵ为首次从猫爪草中分离得到。     

Anti-inflammatory properties of phenolic lactones isolated from Caesalpinia paraguariensis stem bark

Ethnopharmacological relevance

Caesalpinia paraguariensis (D. Parodi) Burkart stem bark infusion (CPBI) is traditionally used in Argentina because their “vulnerary” properties.

Aim of the study

CPBI was studied throughout bio-guided purification procedures conducted by in vitro biological assays in order to isolate the main bioactive compounds.

Material and methods

Anti-inflammatory activity was assessed by enzyme inhibition assays of Hyaluronidase (Hyal) and inducible Nitric Oxide Synthase (iNOS). The antioxidant properties were evaluated by DPPH free radical scavenging assay, lipid peroxidation inhibition assay on erythrocyte membranes, and a cell-based assay that included the fluorescent probe (DCFH-DA) for indicating reactive oxygen species (ROS) generation. Bioactive compounds were purified by chromatographic methods and their structures elucidated using spectroscopic methods (ESI-MS and 1D/2D-¹H/¹³C-NMR).

Results

Four main bioactive compounds were isolated from CPBI: ellagic acid (1), 3-O-methylellagic acid (2), 3,3'-di-O-methylellagic acid (3) and 3,3'-di-O-methylellagic-4-β-D-xylopyranoside (4). These were bioactive at concentrations in which are present in CPBI, being compounds 2 and 3 the best enzyme inhibitors of Hyal and iNOS, reaching the 90% inhibitory concentration (IC₉₀) values ranging from 2.8 to 16.4 μM, that are better than that of the positive controls, aspirin (IC₉₀: no reached) and aminoguanidine (IC₉₀: 20.2 μM) respectively. Compounds 2 and 3 were also better scavengers for lipoperoxides than butylated hydroxytoluene (BHT), reaching the 90% effective concentration (EC₉₀) at 1.2–4.5 μg/ml, and for DPPH radical (2.5–7.3 μg/ml); moreover compounds were able to exert its scavenging action on intracellular ROS. Structural features relevant to the biological activities are discussed.

Conclusions

This work provides scientific validity to the popular usage of CPBI.     

中药创新药物的研发探讨

作者围绕着中药创新药物的研制的相关问题进行了探讨,认为中药创新药物研制的切入点应重点放在中医临床;组分组合和成分组合创新药物的研究是一个重要的研究方向,但组分组合和成分组合不能替代饮片配伍;组分配伍以中医学理论为指导,其目标是能够按照中医理论辨证用药,并且具有较高的安全性,临床适应证明确且针对性好,成分及作用机制相对清楚,质量稳定可控,能够产业化推广。组分配伍模式有从有效方剂的饮片配伍深入到组分配伍,直接组分配伍及单味药的标准组分配伍几类,组分配伍的作用方式仍然是多组分、多靶点、整体综合调节。中药应该制定中药特色的质量标准,不应该一味追求和迎合国际标准;中药创新药物的服务对象应重点放在中国的老百姓。     

Antibacterial compounds from the mushroom Ganoderma colossum from Nigeria

Three colossolactones (colossolactone E, colossolactone B and 23-hydroxycolossolactone E) were isolated and characterized from an n-hexane:dichloromethane (2:7) extract of Ganoderma colossum using chromatographic techniques. The antimicrobial activity of the three compounds was then tested against Gram-positive and Gram-negative bacteria. The activity was evaluated by the thin-layer chromatography agar overlay method. The results showed that colossolactone E and 23-hydroxycolossolactone E were active against Bacillus subtilis and Pseudomonas syringae. Colossolactone B was not active against the bacteria. Their structures were elucidated by spectroscopic methods. Potency of the compounds against bacteria tested supports the use of this mushroom in therapeutic medicine.     

Characterization of the anticancer properties of monoglycosidic cardenolides isolated from Nerium oleander and Streptocaulon tomentosum

Aim of the study

For identification of the active constituents we investigated the anticancer activity of cardenolides from Streptocaulon tomentosum Wight & Arn. (Asclepiadaceae) and from Nerium oleander L. (Apocynaceae) which are both used against cancer in the traditional medicine in their region of origin.

Material, methods and results

The antiproliferative activity of cardenolides isolated from roots of Streptocaulon tomentosum (IC₅₀ < 1-15.3 μM after 2 days in MCF7) and of cardenolide containing fractions from the cold aqueous extract of Nerium oleander leaves (“Breastin”, mean IC₅₀ 0.85 μg/ml in a panel of 36 human tumor cell lines), their influence on the cellular viability and on the cell cycle (block at the G2/M-phase or at the S-phase in tumor cells, respectively) were determined using different cell lines. The murine cell line L929 and normal non-tumor cells were not affected. Bioactivity guided fractionation of Breastin resulted in the isolation of the monoglycosidic cardenolides oleandrine, oleandrigeninsarmentoside, neritaloside, odoroside H, and odoroside A (IC₅₀-values between 0.010 and 0.071 μg/ml).

Conclusions

The observed anticancer activities of extracts and isolated cardenolides are in agreement with the ethnomedicinal use of Streptocaulon tomentosum and Nerium oleander. The most active anticancer compounds from both species are monoglycosidic cardenolides possessing the 3β,14β-dihydroxy-5β-card-20(22)-enolide structure with or without an acetoxy group at C-16. The results indicate that the cytotoxic effects are induced by the inhibition of the plasma membrane bound Na⁺/K⁺-ATPase.