Simultaneous screening, identification, quantitation, and activity evaluation of six acetylcholinesterase (AChE) inhibitors in Coptidis Rhizoma by online UPLC-DAD coupled with AChE biochemical detection.

Acetylcholinesterase (AChE) inhibition is a common treatment in the early stages of dementia and neurodegenerative diseases, including Alzheimer's disease (AD). Coptidis Rhizoma (CR), a traditional Chinese medicine (TCM), contains numerous isoquinoline alkaloids that substantially inhibit AChE played neuroprotective effects in the treatment of cognitive diseases. We established a method using an ultra-high-performance liquid chromatography-diode array detector coupled with AChE biochemical detection (UPLC-DAD-AChEBCD) to screen and identify AChE inhibitors (AChEIs), measure AChEIs content and activity, and evaluate the quality of CR derived from different plant species and growth year. The chromatographic fingerprint and AChEIs activity profiles of CR were simultaneously obtained by UPLC-DAD-AChEBCD, and six alkaloids including groenlandicine, coptisine, epiberberine, jatrorrhizine, berberine, and palmatine, were identified by UPLC-electrospray ionization-tandem mass spectrometry. Data analysis based on AChEIs content and total activity of 12 batches of CR indicated differences among different species and growth year. Therefore, the online method could be used to rapidly identify AChEIs in complex matrixes and screen potential agents for neurodegenerative prevention and treatment, as well as provide information for the identification and quantitation of active markers directly associated with herbal medicine quality.

New steroidal alkaloid and furostanol glycosides isolated from Solanum lyratum with cytotoxicity.

Two previously undescribed steroidal compounds, 16, 23-epoxy-22, 26-epimino-cholest-22(N), 23, 25(26)-trien-3ß-ol-3-O-ß-D-glucopyranosyl-(1→2)-ß-D-glucopyranosyl-(1→4)-ß-D-galactopyranoside (1) and 26-O-ß-D-glucopyranosyl-(25R)-5α-furost-20(22)-en-3ß, 26-diol (2), together with 7 known ones including 26-O-ß-D-glucopyranosyl-(25R)-5, 20(22)-dien-furost-3ß, 26-diol (3), (25R)-5-en-spirost-3ß-ol-O-ß-D-glucopyranosyl-(1→4)-[α-L-rhmanopyranosyl-(1→2)]-ß-D-galactopyranoside (4), funkioside D (5), aspidistrin (6), tigogenin-3-O-ß-D-lucotrioside (7), desglucolanatigonin II (8), and degalactotigonin (9), were isolated from Solanum lyratum Thunb. Their cytotoxic activities were tested in two cancer cell lines by MTT method. One of the steroidal glycosides (6) showed significant cytotoxic activity against gastric cancer SGC7901 and liver cancer BEL-7402 cells.

The complete mitochondrial genome sequence of Erpobdella octoculata (Hirudinea: Arhynchobdellida: Erpobdellidae).

Erpobdella octoculata (Linnaeus, 1758; Hirudinea: Arhynchobdellida: Erpobdellidae) is a very common and morphologically variable macrophagous predators of aquatic invertebrates. Here we determined the complete mitochondrial DNA (mtDNA) sequence of this species, as the first representative of the suborder Erpobdelliformes. This genome is 14,407 bp in length with an A + T content of 71.55%, containing 37 typical animal mitochondrial genes and a non-coding region (NCR). It has high AT content and the same gene arrangement pattern as those of typical annelids. The complete mtDNA sequence of E. octoculata provides useful genetic markers for identification, ecological and evolutionary studies of leeches.

[Sequence analysis of rDNA-ITS gene in medicinal leeches and their closely related species (Euhirudinea:Arhynchobdellida)].

OBJECTIVE: To explore the phylogenetic relationships among medicinal leeches and their closely related species by comparing their ITS gene fragments. METHODS: 8 different leech species, including Whitmania pigra, Whitmania acranulata, Whitmania laevis, Hirudo nipponia, Poecilobdella manillensis, Poecilobdella javanica, Poecilobdella hubeiensis and Erpobdella octoculata were collected as the materials for study. After the total DNA of all samples was extracted, amplified by PCR and cloned, their ITS sequence, AT content, variable sites and evolutionary relationships were analyzed. RESULTS: The length of ITS was 948-981 bp with 210 variable sites and 43 inserts on/deletion. The average G + C content was high up to 56.0%-57.8%. The aligned ITS sequence demonstrated that their ITS genes were relatively conserved in species. Data analysis showed that the relationship between E. octoculata and P. javanica was closer than others, their genetic distance was only 0.007. Through the phylogenetic tree analysis, three species of Haemopidae and four species of Hirudinidae were obviously clustered into two branches. CONCLUSION: The results support the taxonomic status of Haemopidae and Hirudinidae, and provide sufficient evidences that Erpobdella octoculata and Poecilobdella javanica should not be assigned to two genera, of which the taxonomic system should be further researched.