Anoectochilus roxburghii polysaccharides exhibit notable hepatoprotective effects, but the underlying substance basis and mechanisms remain unknown. In this study, four new polysaccharides named ARP-1a, ARP-1b, ARP-2a and ARP-2b, were isolated from A. roxburghii. Their structural characteristics were systematically analyzed using HPGPC, HPLC, GC–MS, IR and NMR analysis. ARP-1a, the leading polysaccharide isolated from A. roxburghii, was further evaluated for its hepatoprotective effects on acute liver injury mice induced by CCl4. ARP-1a significantly reduced the serum ALT, AST, TNF-α, IL-1β and IL-6 levels, liver MDA content, and increased the SOD and CAT activities and GSH level in liver. H&E staining revealed that ARP-1a pretreatment could markedly relieve liver injury. Further mechanism exploration indicated that ARP-1a could relieve CCl4-induced oxidative damage through activating the Nrf2 signaling. In addition, metabolomics, lipidomics and 16S rRNA amplicon sequencing were used to elucidate the underlying mechanisms of ARP-1a. Multi-omics analysis indicated that ARP-1a exerted hepatoprotective effect against CCl4-induced acute liver injury by regulating lipid metabolism and modulating the gut microbiota. In conclusion, the above results suggest that ARP-1a can be considered a promising and safe candidate for hepatoprotective drug, as well as a potential prebiotic for maintaining intestinal homeostasis and promoting human intestinal health.

Ethnopharmacological relevance: Anoectochilus elatus Lindl. was traditionally used for pain treatment and Gooderoside A (GA) was regarded as its principal constituent. Aim of the study: To investigate whether GA can be responsible for the antinociceptive activity of A. elatus and explore its underlying mechanism. Materials and methods: Acetic acid-induced abdominal writhing and tail flick tests were employed to evaluate the antinociceptive activity of ethanolic extract of A. elatus (EEA) and GA. Formalin test was used to ascertain the antinociceptive pattern of GA. Entobarbital sodium induced sleep test was adopted to exclude its hypnotic effect, while open-field test was performed to rule out its motor impairment effect. Chronic constriction injury (CCI)induced neuropathic pain in rats was developed to evaluate its efficacy on neuropathic pain, and BV-2 cells were used to explore the underlying mechanism. Results: EEA and GA, significantly inhibited chemical and thermal nociception. GA suppressed nociception in formalin test in both phase I and II, whereas methylene blue and L-NAME partially reversed its efficacy. GA located inner and slightly blocked sodium channel current, and did not show any hypnotic effect or motor impairment effect. Crucially, GA markedly attenuated chronic neuropathic pain in rats, inhibited the phosphorylation of IRAK4, IRAK1 and TAK1, and suppressed MAPKs pathway in BV-2 cells. Conclusion: GA relieved acute and chronic pains in vivo. The mechanism of action involves the blocking of NO/ cGMP and IRAK4/IRAK1/TAK1 pathways. These results suggested GA may be a promising candidate for antinociceptive drug development.

Anoectochilus roxburghii polysaccharides exhibit notable hepatoprotective effects, but the underlying substance basis and mechanisms remain unknown. In this study, four new polysaccharides named ARP-1a, ARP-1b, ARP-2a and ARP-2b, were isolated from A. roxburghii. Their structural characteristics were systematically analyzed using HPGPC, HPLC, GC-MS, IR and NMR analysis. ARP-1a, the leading polysaccharide isolated from A. roxburghii, was further evaluated for its hepatoprotective effects on acute liver injury mice induced by CCl4. ARP-1a significantly reduced the serum ALT, AST, TNF-alpha, IL-1 beta and IL-6 levels, liver MDA content, and increased the SOD and CAT activities and GSH level in liver. H&E staining revealed that ARP-1a pretreatment could markedly relieve liver injury. Further mechanism exploration indicated that ARP-1a could relieve CCl4-induced oxidative damage through activating the Nrf2 signaling. In addition, metabolomics, lipidomics and 16S rRNA amplicon sequencing were used to elucidate the underlying mechanisms of ARP-1a. Multi-omics analysis indicated that ARP-1a exerted hepatoprotective effect against CCl4-induced acute liver injury by regulating lipid metabolism and modulating the gut microbiota. In conclusion, the above results suggest that ARP-1a can be considered a promising and safe candidate for hepatoprotective drug, as well as a potential prebiotic for maintaining intestinal homeostasis and promoting human intestinal health.

The seeds of Vitex negundo have been used for inflammation‐related disease treatment in traditional medicine. This study focused on the anti‐osteoarthritis (OA) effects of the total lignans of V. negundo seeds (TOV) in monosodium iodoacetate‐induced osteoarthritis rats and its pharmacokinetic properties, as well as the effects and potential mechanism of its main components VN1 (6‐hydroxy‐4‐(4‐hydroxy‐3‐methoxy‐phenyl)‐3‐hydro‐xymethyl‐7‐methoxy‐3,4‐dihydro‐2‐naphthaldehydeb) and VN2 (vitedoin A) on receptor activator of NF‐κB ligand (RANKL)‐induced osteoclast differentiation in bone marrow macrophages (BMMs). TOV significantly attenuated osteoarthritis, leading to an increase in pain thresholds, improvement of knee articular cartilages and chondrocytes loss, and decreased total joint scores and serum levels of TNF‐α, interleukin‐1β (IL‐1β), and prostaglandin E2 (PGE2) in osteoarthritis rats. The half‐time (T1/2) was 2.82 h and 1.33 h, and the bioavailability was 15.34%–21.89% and 16.29%–22.11%, for VN1 and VN2, respectively. VN2, rather than VN1, remarkably inhibited tartrate‐resistant acid phosphatase (TRAP) activity, reduced the number of TRAP‐positive multinuclear cells, diminished the formation of actin ring, and decreased mRNA levels of cathepsin K (CTSK), TRAP, nuclear factor of activated T cell 1 (NFATc1), and osteoclast‐associated receptor, as well as downregulated protein levels of p‐ERK (phosphorylated extracellular signal‐regulated kinase), TRAP, CTSK and NFATc1 in BMMs. These findings suggest TOV has promising therapeutic potential for OA treatment and VN2, in particular, attenuates osteoclast differentiation by suppressing ERK/NFATc1 signaling and actin ring, mainly accounting for the anti‐OA efficacy of TOV.

Background: Trichoderma species are rich source of bioactive secondary metabolites. In the past decades, a series of secondary metabolites were reported from different Trichoderma fungi, among which ter-penoids possessing versatile structural diversities and extensive pharmacological activities are one of the particularly important categories.Aim of Review: The review aims to summarize the terpenoids isolated from Trichoderma species regarding their structural diversities, biological activities, and promising biosynthetic potentials.Key Scientific Concepts of Review: So far, a total of 253 terpenoids, including 202 sesquiterpenes, 48 diter-penes, 2 monoterpenes and 1 meroterpenoid, were isolated and identified from Trichoderma species between 1948 and 2022. Pharmacological investigations of Trichoderma terpenoids mainly focused on their antibacterial activities, antifungal activities, inhibitory activities on marine plankton species and cytotoxic activities, indicating that Trichoderma species are important microbial agents for drug discovery and environmentally friendly agrochemicals development. Intriguing chemistry and enzymology involved in the biosynthesis of Trichoderma terpenoids were also presented to facilitate further precise genome mining-guided novel structure discovery. Taken together, the abundance of novel skeletons, bioactivities and biosynthetic potentials presents new opportunities for drug and agrochemicals discov-ery, genome mining and enzymology exploration from Trichoderma species. The work will provide refer-ences for the profound study of terpenoids derived from Trichoderma, and facilitate further studies on Trichoderma species in the areas of chemistry, medicine, agriculture and microbiology.& COPY; 2023 The Authors. Published by Elsevier B.V. on behalf of Cairo University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

实验教学在中药炮制课程中占据重要地位,是将理论知识转化为实践经验的重要手段.随着近年来互联网技术以及人工智能技术的迅速发展,虚拟仿真技术在实验教学方面应用广泛,其交互性、构想性、开放性等特点能够弥补传统教学的不足,优化教学资源.本文分析了中药炮制实验教学现状及存在的不足,研讨虚拟仿真技术在中药炮制实验教学中的优势,以期提高课程教学质量,为炮制实验教学改革提供参考.

目的 研究从金线莲Anectochilus roxburhii中分离的一株曲霉属内生真菌Aspergillus sp.J218的次生代谢产物.方法 利用Sephadex LH-20凝胶、硅胶和高效液相制备等色谱法从J218固体发酵产物的乙酸乙酯部位分离化合物,采用波谱方法对化合物进行结构鉴定.结果 从金线莲内生真菌Aspergillus sp.J218的次生代谢产物中共分离得到 10个化合物,分别鉴定为 kotanin(1)、flavasperone(2)、aurasperone B(3)、fonsecinone B(4)、fonsecinone D(5)、tensidol A(6)、fonsecinone A(7)、fonsecinone C(8)、aurasperone A(9)和fonsecinone F(10).结论 从金线莲内生真菌曲霉属Aspergillus sp J218次生代谢产物分离鉴定多为二聚萘并吡喃酮类化合物,提示该菌株含有丰富的二聚化合成酶,可为后续探索该菌株中二聚萘并吡喃酮类化合物的生物合成途径提供线索.

A new species of Anoectochilus (Orchidaceae) from Guangxi, China, A.zhongshanensis, is described here, which was identified based on phylogenetic studies adopting combined plastid markers (rbcL-matK-trnL-F), morphological observation and chemical analysis. Molecular phylogenetic results support the systematic status of A.zhongshanensis as a new species in Anoectochilus genus. Morphologically, this new species is similar to A.zhejiangensis and A.malipoensis, but differs by its characteristic labellum and column, including the hastate or scalpel-shaped lobes of epichile, forward curved and pinnately divided cristate lobes at both sides of the mesochile and inverted triangle column wings. Furthermore, HPLC-ELSD analysis of these three species revealed the interesting chemotaxonomic difference that the principle and characteristic lactone glycoside in this new species was kinsenoside, rather than its diastereoisomer, goodyeroside A, a major glycoside in A.zhejiangensis and A.malipoensis.

Anoectochilus elatus is a new record species from Yunnan province in China discovered by our group in 2018, used in folk as the most popular Anoectochilus species A. roxburghii for medicinal and culinary purposes. The crude polysaccharide of Anoectochilus elatus (AEP) exhibited significant antinociceptive effects against both chemical and thermal nociception in vivo. Bio-guided isolation identified GJXL-1 as the leading analgesic polysaccharide in AEP. Detailed structural analyses rationalized GJXL-1 (molecular weight: 10.3 kDa) as an α-D-1,4-linked glucan unexpectedly branched at O-3, and O-6 position. GJXL-1 dose-dependently suppressed acetic acid-induced writhing in mice and decreased the serum levels of NO, IL-6 and TNF-α, which also repressed the licking times in both the first and second phases in formalin test. Furthermore, only L-nitroarginine partly reversed the analgesic activity of GJXL-1, indicating that GJXL-1's efficacy was partially mediated by NO regulation, possibly through inhibiting IRAK4/TAK1/NF-κB signaling pathway, and modulating gut microbiota and short-chain fatty acids production. In addition, the motor impairment and hypnotic effects of GJXL-1 were excluded. Our study suggests that GJXL-1 can be regarded as a promising and safe drug candidate for diverse pain disorders, and also a promising prebiotic candidate to maintain intestinal homeostasis and promote human gut health.

中药鉴定学与中药炮制学是中药学专业的两门必修主干课程,将这两门课程进行联合教学,通过设置教学内容、教学实施及教学评价等,把中药鉴定与炮制的知识与技能相互引用、相互印证,优化了课程教学与实验资源,增强了学员中药鉴定与炮制的相关技能以及运用鉴定知识分析炮制变化的能力.联合教学具有博采教学资源、贴近中药炮制实用需求、扩展中药鉴定知识范围、增强学员学习兴趣、提高学员专业技能的现实意义.

目的 制定刺梨药材质量标准,用于刺梨的生产、监督、流通及使用等各环节的质量控制.方法 参照《中国药典》通用检测方法及相关指导原则分别制定刺梨的水分、总灰分、浸出物限度;采用特征图谱以及薄层色谱法控制刺梨整体质量.结果 刺梨特征图谱中应呈现6个特征峰,并应与对照药材参照物色谱峰中的5个特征峰保留时间相对应,另有1峰应与加入的异槲皮苷对照品参照物峰保留时间相对应.水分不得过14.0％,总灰分不得过5.0％,醇溶性浸出物不少于20.0％.结论 实验所建立的刺梨质量标准符合国家有关中药质量标准制定要求,能够以标准的形式对刺梨的质量进行控制.

目的 研究中药海龙对D-半乳糖诱导衰老小鼠学习记忆损伤的影响及其作用机制.方法 采用高效液相色谱(HPLC)技术测定海龙中抗学习记忆损伤有效成分二十二碳六烯酸(DHA)含量;腹腔注射D-半乳糖(D-gal)制备衰老小鼠模型,以Morris水迷宫实验及蛋白免疫印迹法等,对小鼠学习记忆能力、海马组织氧化损伤相关生化指标及蛋白表达进行检测,探究海龙对衰老小鼠学习记忆损伤的保护作用及其机制.结果 HPLC结果显示,海龙中DHA含量为7.7613 mg/g(以生药计),约占总成分的47％;Morris水迷宫实验结果显示,海龙可减少学习记忆损伤小鼠逃避潜伏期时间,增加小鼠目标象限游泳时间、游泳路程占比、穿台次数,改善小鼠学习记忆损伤;此外,海龙可激活衰老小鼠海马组织AKT/FOXO1/SOD2信号通路,上调氧化应激通路相关蛋白表达,降低衰老小鼠海马组织氧化损伤程度改善小鼠学习记忆损伤.结论 本研究发现海龙富含二十二碳六烯酸,具有改善D-半乳糖诱导衰老小鼠学习记忆损伤的作用,并初步阐明其作用机制与抗氧化相关,为中药海龙抗学习记忆损伤研究提供了实验依据.

Two new polysaccharides, AZP-1a and AZP-1d, with molecular weights of 3.41 × 104 and 4568 Da, respectively, were extracted from Anoectochilus zhejiangensis and purified by column chromatography. Their structural characteristics were systematically explored and results indicated AZP-1a and AZP-1d shared a similar backbone consisted of→4)-Galp-(1→, →4)-Glcp-(1→, and →4,6)-Glcp-(1→, with a different terminal residue of Manp-(1 → and Glcp-(1→, respectively. In vivo experiments showed that the crude polysaccharide of A. zhejiangensis (AZP) exhibited significant hepatoprotective effects, decreasing the serum levels of ALT, AST and LDH in CCl4-treated mice, reducing MDA content, promoting SOD and CAT activities, and increasing GSH level in liver. Further in vitro investigation exhibited that AZP, AZP-1a and AZP-1d effectively protected liver cells against CCl4-stimulated oxidative damage, while AZP-1a and AZP-1d functioned mainly through the activation of Nrf2 signaling pathway. Our results suggest that A. zhejiangensis polysaccharides can be applied as a potential resource for the development of hepatoprotective drugs.

目的 对目前市场上流通的中药材海马及其混淆品微性状鉴别研究,为海马的鉴别研究提供依据.方法 利用性状鉴别法对收集到的海马及其混淆品进行鉴别;运用中药微性状鉴定法对11种不同品种海马的鳃盖和冠应用体式显微镜拍摄,运用景深合成技术合成1张高清微性状照片进行鉴别研究.结果 不同品种海马在鳃盖的嵴纹以及头冠有较明显的区别.结论 微性状鉴别法能够有效将市场上流通的不同品种的海马进行区分,并且具有简单快捷的特点.

Prostate cancer is the second most common malignant cancer in males. It involves a complex process driven by diverse molecular pathways that closely related to the survival, apoptosis, metabolic and metastatic characteristics of aggressive cancer. Prostate cancer can be categorized into androgen dependent prostate cancer and castration-resistant prostate cancer and cure remains elusive due to the developed resistance of the disease. Natural compounds represent an extraordinary resource of structural scaffolds with high diversity that can offer promising chemical agents for making prostate cancer less devastating and curable. Herein, those natural compounds of different origins and structures with potential cytotoxicity and/or in vivo anti-tumor activities against prostate cancer are critically reviewed and summarized according to the cellular signaling pathways they interfere. Moreover, the anti-prostate cancer efficacy of many nutrients, medicinal plant extracts and Chinese medical formulations were presented, and the future prospects for the application of these compounds and extracts were discussed. Although the failure of conventional chemotherapy as well as involved serious side effects makes natural products ideal candidates for the treatment of prostate cancer, more investigations of preclinical and even clinical studies are necessary to make use of these medical substances reasonably. Therefore, the elucidation of structure-activity relationship and precise mechanism of action, identification of novel potential molecular targets, and optimization of drug combination are essential in natural medicine research and development.

Chemical investigation on the solid rice culture of Chaetomium globosum D38, an endophytic fungus derived from Salvia miltiorrhiza, has afforded two new 19,20-seco-chaetoglobosins, salchaetoglobosins A (1) and B (2), along with three known analogues, chaetoglobosins E (3), Fex (4), and Vb (5). Their structures and absolute configurations were elucidated by a set of spectroscopy and single-crystal X-ray crystallography. Compounds 1-5 were evaluated for their cytotoxic activities against HCT-116 (colorectal carcinoma) and PC3 (prostate cancer) cells, as well as the NO production inhibitory effects in LPS-stimulated RAW264.7 cells.

Ethnopharmacological relevance: Osteoporosis and Alzheimer's disease (AD) are both senile diseases, which are closely related to oxidative stress. Bajitianwan (BJTW) is a classic Chinese formulation consisting of seven herbal drugs: the root of Morinda officinalis F.C.How., root and rhizome of Acorus tatarinowii Schott, the root bark of Lycium chinense Mill., the sclerotium of Poria cocos (Schw.) Wolf, the root of Polygala tenuifolia Willd., sclerotium with host wood of Poria cocos (Schw.) Wolf and root and rhizome of Panax ginseng C. A. Mey. BJTW has been used for the treatment of osteoporosis and AD for hundreds of years. Aim of the study: This study aimed to investigate the protective effects of BJTW in the amelioration of memory impairment and bone loss induced by D-galactose and to explore the underlying mechanism. Materials and methods: The aging model was established in male Wistar rats by subcutaneous injection of D-galactose (100 mg/kg), and the rats were treated with huperzine-A, alendronate sodium, or the aqueous extract of BJTW for 4 months. Cognitive performance was evaluated with the Morris water maze. Rat femurs were scanned using microcomputed tomography to obtain three-dimensional imagery of bone microstructure. The impact of D-galactose on the expression of Forkhead box O1 and superoxide dismutase 2 in femur tissue was also evaluated. Results: For the model group, BJTW treatment significantly reduced the latency time for finding the target platform in the directional swimming test and increased time spent swimming in the target quadrant with the probe test. Additionally, BJTW treatment alleviated D-galactose-induced bone loss through regulation of levels of alkaline phosphatase, osteocalcin, osteoprotegerin, and receptor activator of nuclear factor kappa B ligand. Furthermore, BJTW treatment increased catalase and glutathione peroxidase levels in serum, reduced malondialdehyde content in hippocampus, and upregulated expression of Forkhead O1, which upregulated superoxide dismutase 2 in the femur. Conclusions: BJTW had positive effects on age-related memory impairments and bone loss. It may be a promising antioxidant candidate for treatment of Alzheimer's disease and osteoporosis.