BACKGROUND:Aspongopus chinensis Dallas holds both medicinal and culinary significance in China. Notably, in regions such as Guizhou and Yunnan, it has been traditionally used as an ethnic remedy for treating various conditions, including stomach coldness, pain, kidney deficiencies, and impotence, among other ailments. This study aims to explore the chemical constituents and anticoagulant activity of A. chinensis. METHODS:Ultraperformance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was employed for the UPLC fingerprint analysis. The isolated compounds underwent in vitro assays to their evaluate anticoagulant properties and effects on protein fibril activity. Network pharmacology was utilized to predict potential anticoagulant targets. Furthermore, animal experiments were conducted to measur coagulation factors and assess the in vivo anticoagulant activity. RESULTS:Our findings indicate that 10 compounds were identified through UPLC-Q-TOF-MS analysis, with four compounds (uracil; 6-hydroxyquinolinic acid; 1,4-dihydro-4-oxoquinoline-2-carboxylic acid; delicatuline B) were isolated and identified from the n-butanol extract of A. chinensis. Furthermore, principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (PLS-DA) of the UPLC fingerprint data revealed significant differences between A. chinensis and its similar insects. Notably, 1,4-dihydro-4-oxoquinoline-2-carboxylic acid has been identified as a potential standard reference substance for content determination. The isolated compounds showed anticoagulant properties, and the mRNA expression levels of MMP9 and PTGS2 were significantly reduced in LPS-induced RAW264.7 cells, further supporting the network pharmacology analysis. Animal experiments confirmed the potent anticoagulant effects of A. chinensis, likely associated with the intrinsic coagulation pathway and the inhibition of platelet aggregation. CONCLUSION:This research provides a quality assessment method for A. chinensis, and has also demonstrated its anticoagulant functions and substance basis.

ETHNOPHARMACOLOGICAL RELEVANCE:Selaginella moellendorffii Hieron. has been used as ethnic drug for chronic inflammation treatment. Biflavonoids represent a crucial class of bioactive compounds recognized for their potent anti-inflammatory activity in S. moellendorffii (SM). However, the effective components, targets, and pathways that SM in anti-inflammasome remain unclear. AIM OF THE STUDY:Therefore, this study initially evaluated the effective components of SM and explored the underlying mechanisms. MATERIALS AND METHODS:Firstly, a series of biflavonoids were isolated from SM, and then all compounds were evaluated for their anti-inflammatory ability in the THP-macrophages co-stimulated with lipopolysaccharide (LPS) and NLRP3 inflammasome inducers. Secondly, transcriptomic analysis and metabolomics analysis revealed the differential genes and metabolites associated with effective components treatment. Finally, molecular docking of effective components with NLRP3 was performed and western blotting was performed in order to determine the expression of related proteins. RESULTS:Overall, eleven biflavonoids were successfully isolated from SM. Particularly, F7 exhibited the most potent inhibitory effect against NLRP3 inflammasome-mediated cytokines levels, cell membrane integrity and Ca2+ influx. Transcriptomic studies demonstrated that the differential genes (DEGs) were mainly enriched in NF-κB signaling pathway and NOD-like receptor signaling pathway. Metabolomics studies that the metabolites were mainly involved the pyrimidine metabolites. Further validation analysis manifested that F7's significant downregulation of NLRP3 inflammasome-related genes and proteins expression (P < 0.05, P < 0.01), encompassing both priming (NLRP3, TNF-α, p-p65/p65) and activation stages (IL-1β, IL-18, Caspase-1, GSDMD-N/GSDMD). Moreover, NLRP3 knockdown attenuated F7-mediated inhibition of pyroptosis. Finally, in silico results showed that F7 exhibited promising predicted binding affinity towards NLRP3. CONCLUSIONS:Collectively, these findings revealed an anti-inflammatory material basis for SM and confirmed F7 as a potent inhibitor of pyroptosis by suppressing NF-κB/NLRP3 Pathway.

采用体视显微镜对藏药独一味的根、茎、叶和果实微性状进行观察,利用生物显微镜观察切片组织构造及粉末显微鉴别特征,扫描电镜进一步观察叶片表面结构特征.藏药独一味药材的微性状特征包括根、茎、叶、果实,而茎粗短,果实细小且易脱落,地上部分鉴定特征主要为叶片和果序,叶片表皮细胞中可见气孔、非腺毛和腺鳞等.地上部分粉末鉴定特征包括非腺毛、叶肉细胞、气孔和纤维等,根的鉴别特征包括针晶和导管等.本研究通过对独一味药材不同部位微性状和显微进行系统研究,归纳了独一味药材性状和显微的鉴别要点,可以为独一味药材鉴别提供参考依据.

Ethnopharmacological relevance: Selaginella moellendorffii (SM) has been applied as an ethnic drug to treat conditions such as osteoporosis, idiopathic thrombocytopenic purpura, and chronic inflammation. It is known to be rich in flavonoids, including apigenin glycosides and unique elements of bioflavonoids.Aim of the study: To investigate estrogen-like constituents of SM and the possible mechanism.Materials and methods: We identified the main components in liquid chromatography and liquid chromatographymass spectrometry. The estrogenic effects were examined using a recombinant yeast screening assay, an E-screen cell proliferation assay, and an in vivo uterotrophic assay.Results: Flavonoid glycosides extract, some flavonoid glycosides, and apigenin showed estrogen agonistic activity in the yeast screening assay. They also induced cell proliferation in estrogen receptor-positive (ER+) cells but not in estrogen receptor-negative (ER-) cells. Consistently, the protein expression of ER alpha, phosphorylation protein kinase B (p-AKT), phosphatidylinositol 3 kinase (PI3K), phosphorylation mammalian target of rapamycin (pmTOR), phosphorylation 38,000-Da protein (p-P38), and phosphorylation extracellular-regulated kinase 1/2 (pERK1/2) elevated following treatment with flavonoid glycoside extract (P < 0.01 or P < 0.05). These effects could be blocked by ER antagonist or ER alpha antagonist but not be blocked by ER beta antagonist. In vivo assay, flavonoid glycoside extract could significantly increase body weight, serum estradiol level, uterine wet weight, alter uterine morphology, and promote ER alpha protein expression (P < 0.01 or P < 0.05). Conclusions: ER alpha induction via mitogen-activated protein kinases (MAPK) and PI3K/Akt/mTOR pathways might be the possible mechanism underlying the phytoestrogen effect of SM, and the flavonoid glycosides might be the critical estrogenic constituents.

基于文献整理,分析药用菊花的历史发展过程,通过地理分布特点及主要类型形态特征分析、比较,探讨药用菊花起源和演化规律.结果表明,药用菊花存在由野生到栽培的发展过程,宋代以前药用种类均来源于野生菊类.栽培菊出现于唐代,宋代是药用菊花形成、发展的关键时期,明清后逐渐发展成为现代药用类型.菊花是由毛华菊、野菊等多种菊属植物杂交后经人工选育、引种形成的不育类型,河南南阳及大别山区是菊花的重要起源地.不同种类菊花演化进程存在差异,小洋菊、大洋菊、福白菊及滁菊属原始平瓣菊类型;亳菊、济菊、祁菊源于怀菊,为后期进化类型;贡菊为过渡类型.菊花的种植类型与规模受商业影响在不断变化,需加强药材质量控制和资源保护,新品种的发现对扩充种质资源具有重要意义.

目的:比较顶空-固相微萃取法(HS-SPME)和水蒸气蒸馏法(SD)对宽叶山蒿叶挥发性成分的提取效果.方法:采用GC-MS技术分别对HS-SPME与SD的提取物进行分析.结果:HS-SPME法提取物共鉴别出46个化合物,主要为石竹烯氧化物、石竹烯等;SD法提取物共鉴别出31个化合物,主要为石竹烯氧化物、异丁子香烯等.两种方法共鉴定出挥发性成分54种,共有成分23种,SD法特有成分8个,HS-SPME法特有成分23个.两种方法提取的挥发性成分种类、数量和含量差异较大.结论:HS-SPME-GS-MS法可简便、高效地检测宽叶山蒿叶的挥发性成分,但不能完全替代传统的SD-GC-MS方法,将两种方法联用分析宽叶山蒿叶中挥发性成分能够得到更加全面的信息.

The renin-angiotensin system (RAS) is the primary pathway for regulating blood pressure in the body, and angiotensin-converting enzymes (ACEs) play a crucial role in it. Hirudo nipponia is an invertebrate that contains a variety of active peptides; however, there are no studies on the ACE inhibitory activity of hirudo. In the present study, our aim was to identify the active peptides in hirudo based on active peptide database analysis, unexpectedly filling the gap in hirudo ACE inhibitory activity research. Prep-HPLC was used to separate the part below 3 kD from hirudo. The peptide composition of the isolates was obtained based on Orbitrap LC-MS. The activity of each group of peptides was predicted by the database and the activity was determined by bioassay. Peptides with validation activity were screened through the database. In total, 337 peptides and 18 peptides matching the NCBI leech protein database were identified. All four fractions showed ACE inhibitory activity, and the IC50 was 0.8266, 0.2708, 0.4432, and 0.1764 mg/mL, respectively. Six screened peptides showed good affinity for ACE. This work reveals for the first time that low-molecular-weight peptides from H. nipponia have ACE inhibitory activity, which can provide a new explanation for leech treatment of hypertension.

目的 基于网络药理学和睾丸间质细胞模型探究恩施巴戟对睾丸间质细胞睾酮分泌的影响和潜在作用机制.方法 利用网络药理学和分子对接预测恩施巴戟治疗睾酮缺乏症的主要作用靶点及通路;通过LC-MS/MS技术分析恩施巴戟主要化学成分;构建睾丸间质细胞模型,采用MTT法检测睾丸间质细胞的存活率,采用ELISA法检测睾酮分泌量,采用qRT-PCR检测睾酮合成和凋亡相关基因表达,采用Western blotting检测睾酮合成蛋白表达.结果 网络药理学和分子对接结果显示,恩施巴戟主要通过丝裂原活化蛋白激酶1(mitogen-activated protein kinase 1,MAPK1)、磷脂酰肌醇3-激酶调节亚基1(phosphatidylinositol 3-kinase regulatory subunit 1,PIK3R1)等靶点和环磷酸腺苷(cyclic adenosine monophosphate,cAMP)、MAPK、磷脂酰肌醇 3-激酶(phosphatidylinositol 3-kinase,PI3K)/蛋白激酶 B(protein kinase B,Akt)等信号通路发挥作用,水晶兰苷为其主要有效成分.LC-MS/MS分析和睾丸间质细胞实验结果验证了网络药理学预测分析,恩施巴戟提取物和水晶兰苷均具有促进睾丸间质细胞生长和睾酮分泌的作用,其作用机制与促进类固醇生成急性调控蛋白(steroidogenic acute regulatory protein,StAR)、胆固醇侧链裂解酶(cholesterol side-chain cleavage enzyme,Cyplla1)、B 淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)/Bcl-2 相关 X 蛋白(Bcl-2 associated X protein,Bax)mRNA 表达(P＜0.05、0.01)并促进 StAR 蛋白表达有关(P＜0.01).结论 恩施巴戟具有促进睾酮分泌的作用,水晶兰苷为其主要活性成分,其作用机制与作用于MAPK、PI3K/Akt通路调控睾酮合成有关.

位于大别山区的湖北麻城种植的以"麻城福白菊"为品牌的药用菊花,历史悠久,品质优良,占据全国较大市场份额.麻城福白菊是原农业部农产品地理标志产品、国家市场监督管理总局地理标志证明商标,2012年通过国家药监局中药材GAP认证,2020年入选中欧地理标志协定首批保护名录,成为首批"中欧100+100"地理标志互认互保产品之一,是中国药用菊花唯一入选品种.文章从历史文化溯源和本草考证角度证实,湖北麻城种菊、用菊历史悠久,有确切记载的历史至少可以追溯到南北朝时期(552年前).

在中国,水蛭是一种珍贵的中药,具有很强的药用价值和研究价值.在世界范围内,水蛭的药用价值同样得到了认可,欧洲及埃及、印度都有水蛭治疗疾病的应用.对国内4种和国外9种主要药用水蛭进行介绍,对已发现的30种具有生物活性的水蛭多肽进行综述.这些多肽活性以抗凝活性为主,还涉及蛋白酶抑制、抗炎、水解酶等活性,以期为水蛭研究、应用提供参考.

目的:建立一种能够准确鉴别九香虫及混伪品的特异性PCR方法,解决形态相似动物药材的鉴定问题.方法:收集不同产地九香虫及其常见混伪品8种123份,基于九香虫及其混伪品的细胞色素C氧化酶亚基I(COI)序列差异,利用Clustal W软件进行同源比对并且筛选出特异性单核苷酸多态性(SNP)变异位点,根据变异位点设计特异性鉴别引物,采用3步法进行PCR扩增,并对影响PCR反应体系的反应条件进行考察,实测鉴定了九香虫及混伪品.结果:当模板量30～100 ng,退火温度为55℃,循环次数为35次时,九香虫能扩增出301 bp的单一明亮条带,而其混伪品均不具有此特异性条带.结论:使用特异性引物对九香虫及其混伪品进行验证的结果良好,与传统形态鉴定结果基本一致.因此,通过特异性PCR方法可以快速准确地鉴定正品九香虫.

目的:对不同种类药用菊花的植物形态特征进行比较鉴别,并对其药材质量进行评价.方法:采用形态鉴定法对不同种类叶型、叶长宽比、花序直径、舌状花瓣型及管状花数等指标进行观测;采用HPLC法测定不同类型菊花药材中绿原酸、木犀草苷及3,5-O-二咖啡酰基奎宁酸的含量.结果:小亳菊、济菊、祁菊及小怀菊植株形态特征相近,而小洋菊、大洋菊、福白菊差异明显,叶长宽比范围为1.16～1.54,福白菊最小,而小洋菊最大.福白菊、小洋菊、大洋菊及滁菊舌状花平瓣,管状花多数,花盘明显;小怀菊、小亳菊、济菊及祁菊舌状花为混合瓣型,花序内具多数膜质小苞片,管状花少见或无;贡菊管状花少,舌状花平瓣,未见混合瓣,花序内苞片偶见.不同类型药材中3种成分总含量由高至低依次为:福白菊、滁菊、济菊、贡菊和杭白菊、祁菊、亳菊、怀菊;3,5-O-二咖啡酰基奎宁酸及绿原酸成分为各类型中主要不合格指标,10批次样品中3,5-O-二咖啡酰基奎宁酸含量低于标准限度值,占比22.7％;5批次样品中绿原酸含量低于限度值,占比11.4％.结论:不同种类菊花植物形态特征及内在质量存在差异,该研究为菊花种类鉴定及药材质量评价提供了必要的数据支撑.

目的:为白前的深入研究以及开发利用提供本草学依据.方法:实地走访湖北新洲、团风和江西庐山白鹿镇梅溪村等当前白前主产地以及中药材市场进行调查,结合文献资料,对白前进行本草考证,梳理总结白前的名称、基原、产地、品质评价、加工及炮制方法、性味功效等方面的沿革.结果 与结论:白前始载于《名医别录》,为常用中药.该药材别名石蓝,又名嗽药等,但历代本草中均以"白前"为正名.古代医家常将白前与白薇混淆使用,缘于宋《本草图经》所附图示之误;后北宋《重修政和经史证类备用本草》完全沿用《本草图经》中对白前和白薇的描述和图示,故从宋代开始,各白薇产地以"叶如柳"的白前误为白薇;至明《救荒本草》开始,医家对前人的错误进行了修正,从叶形态上将"白薇"和"柳叶白前"区分开.至民国,鹅绒藤属及其基原植物分类有较大争议,《植物学大辞典》与《中国药学大辞典》都将白前归类到夹竹桃科白前属下.现代本草书籍对于白前基原记载趋于一致.白前资源从江苏、浙江一带的长江流域沿岸逐步往长江流域中部、南部转移,其与白前生境特点的生长环境发生地理位置变化有关,并与今白前主要栽培产区湖北新洲、团风一带环境一致.白前以根茎粗壮、须根长、无杂质泥土者为佳,但其内在质量还需有效成分分析和药理学证据来佐证.性味功效方面,元代以前,白前皆"味甘",但"微温"还是"微寒"的记载不同,现多记载为"辛、苦,微温".可见,有必要在厘清白前古今基原植物、产地、加工炮制的基础之上,开展化学成分、品质、药效及分子鉴定方面的研究,制定白前商品规格等级标准,以满足其药材交易和流通中按质论价的需求,从而保证临床用药安全.

目的:探讨湿润伤口灵对大鼠创面修复的影响及可能的机制.方法:选取30只大鼠,每只大鼠背部打3个孔来建立皮肤创伤模型,采用自体对照研究,3个孔分别设定为给药组(湿润伤口灵,n=30)、阳性对照组(重组牛碱性成纤维细胞生长因子凝胶,n=30)及模型组(生理盐水,n=30).分别在建模用药后的第1、3、5、7、10、14天,观察各组大鼠创面愈合情况,同时取创面组织样本进行苏木精-伊红染色(hematoxylin-eosin staining,HE染色)观察病理变化,采用实时荧光定量聚合酶链式反应(RT-qPCR)和免疫组化法比较各组创面血管内皮生长因子(vascular endothelial growth factor,VEGF)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)的表达水平.结果:第3、5、7、10、14天,阳性对照组和给药组的创面愈合率均高于模型组(P<0.05);第7、10、14天,给药组创面愈合率均高于阳性对照组(P<0.05).第14天,模型组创面形成肉芽组织,胶原纤维排列不规整,给药组创面明显形成肉芽组织,胶原纤维及大量汗腺细胞和导管增生、毛囊规整排列.给药组和阳性对照组VEGF与TGF-β1平均累计光密度值均高于模型组(P<0.05).第3、7天,给药组和阳性对照组VEGF和TGF-β1的mRNA相对表达量均高于模型组(P<0.05).第14天,给药组和阳性对照组VEGF和TGF-β1的mRNA相对表达量均低于模型组(P<0.05),且给药组和阳性对照组VEGF和TGF-β1的mRNA相对表达量,差异均无统计学意义(P>0.05).结论:湿润伤口灵能通过促进大鼠创面中VEGF和TGF-β1的表达,显著加快大鼠创面组织的愈合.

目的 研究常见矿物类中药粉末的微性状鉴别法.方法 将36种矿物类中药粉末按白色、黄色、灰褐色或黑褐色、红色或紫色分为4类,利用体视显微镜的立体图像处理方法,分别对这几类矿物中药粉末进行微性状观察,比较和总结其鉴别特征,并制作矿物药粉末微性状鉴别检索表.结果 大多数矿物类中药粉末能通过体视显微镜观察到较明显的微性状特征.结论 该方法为矿物类中药粉末的鉴别提供了直观的微性状特征,可作为矿物药的鉴别参考依据.

Background: Activation of nucleotide oligomerization domain-like receptor protein 3 (NLRP3) inflammasome plays a crucial role in gout. Selaginella moellendorffii has been confirmed effective for the treatment of gout in hospital preparations. Flavonoids, such as amentoflavone (AM), are the main active components of this medicine. Purpose: We aimed to investigate the flavonoid extract (TF) and AM's effects on NLRP3 inflammasome in vitro and their preventive effects on gout in vivo. Methods: LC-MS method was employed to investigate the chemical profile of TF. The cellular inflammation model was established by lipopolysaccharide (LPS) or monosodium urate (MSU) stimulation. The cell membrane integrality and morphological characteristics were determined by using Lactate dehydrogenase (LDH) assay kits, propidium iodide (PI) stain, and scanning electron microscopy (SEM). The inflammatory cytokines and NLRP3 inflammasome activation were determined using enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (RT-PCR), immunofluorescence staining, and western blotting. The acute gout mouse model was induced by MSU injection into footpads, and then the paw edema, inflammatory mediators, and histological examination (HE) were analyzed. Results: The main constituents in TF are AM and robustaflavone. In the cellular inflammation model, TF down-regulated the levels of nitric oxide (NO), TNF-α, and LDH, suppressed NLRP3 inflammasome-derived interleukin-1β (IL-1β) secretion, decreased caspase-1 activation, repressed mature IL-1β expression, inhibited ASC speck formation and NLRP3 protein expression. In an acute gout mouse model, oral administration of TF to mice effectively alleviated paw edema, reduced inflammatory features, and decreased the levels of IL-1β in mouse foot tissue. Similarly, the characteristic constituent AM was also able to down-regulated the levels of NO, TNF-α, and LDH, down-regulate the mRNA expression of IL-1β, TNF-α, caspase-1, and NLRP3. Besides, the foot thickness, lymphocyte infiltration, and IL-1β level were also prevented by AM. Conclusion: The results indicated that TF and its main constituent AM alleviate gout arthritis via NLRP3/ASC/Caspase-1 axis suppression.

目的 对灵芝习用品进行生药学鉴别.方法 系统地观察8份灵芝习用品的性状,包括质地、形状、大小、颜色、断面、气味等以及其显微特征,包括皮壳、菌肉、菌管、孢子特征.采用薄层色谱对其化学成分进行初步分析.结果 8份习用品均鉴定为灵芝科灵芝属真菌Ganoderma sp.,外形有一定差异,又具有共同特征.共同特征包括无菌柄,皮壳硬,皮壳菌丝构造呈纵向交织纹理,含多种三萜类成分.薄层色谱显示8份样品的化学成分一致,提示其生物亲缘关系密切.结论 所用方法为灵芝习用品的鉴别和开发利用提供了实验依据.

BACKGROUND:Evodiamine (EVO), an alkaloid extracted from the traditional Chinese medicine Euodia rutaecarpa, plays an important role in the treatment of cancer. This study was performed to clarify the effects of evodiamine in mice tumor model studies.METHODS:Electronic databases and search engines involved China Knowledge Resource Integrated Database (CNKI), Wanfang Database, Chinese Scientific Journal Database (CSJD-VIP), China Biomedical Literature Database (CBM), PubMed, Embase, Web of Science, and ClinicalTrials.gov databases, which were searched for literature related to the antitumor effects of evodiamine in animal tumor models (all until 1 October 2021). The evodiamine effects on the tumor volume and tumor weight were compared between the treatment and control groups using the standardized mean difference (SMD).RESULTS:Evodiamine significantly inhibited tumor growth in mice, as was assessed with tumor volume [13 studies, n=267; 138 for EVO and 129 for control; standard mean difference (SMD)= -5.99; 95% (CI): -8.89 to -3.10; I2 = 97.69%, p ≤ 0.00], tumor weight [6 studies, n=89; 49 for EVO and 40 for control; standard mean difference (SMD)= -3.51; 95% (CI): -5.13 to -3.90; I2 = 83.02%, p ≤ 0.00].CONCLUSION:EVO significantly suppresses tumor growth in mice models, which would be beneficial for clinical transformation. However, due to the small number of studies included in this meta-analysis, the experimental design and experimental method limitations should be considered when interpreting the results. Significant clinical and animal studies are still required to evaluate whether EVO can be used in the adjuvant treatment of clinical tumor patients.