Ajania pacifica (Nakai) K. Bremer and Humphries Extract Limits MYC Expression to Induce Apoptosis in Diffuse Large B Cell Lymphoma.

The proto-oncogene MYC is frequently dysregulated in patients with diffuse large B-cell lymphoma (DLBCL) and plays a critical role in disease progression. To improve the clinical outcomes of patients with DLBCL, the development of strategies to target MYC is crucial. The use of medicinal plants for developing anticancer drugs has garnered considerable attention owing to their diverse mechanisms of action. In this study, 100 plant extracts of flora from the Republic of Korea were screened to search for novel agents with anti-DLBCL effects. Among them, Ajania pacifica (Nakai) K. Bremer and Humphries extract (APKH) efficiently suppressed the survival of DLBCL cells, while showing minimal toxicity toward normal murine bone marrow cells. APKH suppressed the expression of anti-apoptotic BCL2 family members, causing an imbalance between the pro-apoptotic and anti-apoptotic BCL2 members. This disrupted mitochondrial membrane potential, cytochrome c release, and pro-caspase-3 activation and eventually led to DLBCL cell death. Importantly, MYC expression was markedly downregulated by APKH and ectopic expression of MYC in DLBCL cells abolished the pro-apoptotic effects of APKH. These results demonstrate that APKH exerts anti-DLBCL effects by inhibiting MYC expression. Moreover, when combined with doxorubicin, an essential component of the CHOP regimen (cyclophosphamide, doxorubicin, vincristine, and prednisone), APKH synergistically enhanced the therapeutic effect of doxorubicin. This indicates that APKH may overcome drug resistance, which is common in patients with refractory/relapsed DLBCL. To identify compounds with anti-DLBCL activities in APKH, the chemical profile analysis of APKH was performed using UPLC-QTOF/MSe analysis and assessed for its anticancer activity. Based on the UPLC-QTOF/MSe chemical profiling, it is conceivable that APKH may serve as a novel agent targeting MYC and sensitizing drug-resistant DLBCL cells to CHOP chemotherapy. Further studies to elucidate how the compounds in APKH exert tumor-suppressive role in DLBCL are warranted.

Structural divergence and phylogenetic relationships of Ajania (Asteraceae) from plastomes and ETS.

BACKGROUND: Ajania Poljakov, an Asteraceae family member, grows mostly in Asia's arid and semi-desert areas and is a significant commercial and decorative plant. Nevertheless, the genus' classification has been disputed, and the evolutionary connections within the genus have not been thoroughly defined. Hence, we sequenced and analyzed Ajania's plastid genomes and combined them with ETS data to assess their phylogenetic relationships. RESULTS: We obtained a total of six new Ajania plastid genomes and nine ETS sequences. The whole plastome lengths of the six species sampled ranged from 151,002 bp to 151,115 bp, showing conserved structures. Combined with publicly available data from GenBank, we constructed six datasets to reconstruct the phylogenetic relationships, detecting nucleoplasmic clashes. Our results reveal the affinities of Artemisia, Chrysanthemum and Stilpnolepis to Ajania and validate the early taxonomy reclassification. Some of the plastid genes with low phylogenetic information and gene trees with topological differences may have contributed to the ambiguous phylogenetic results of Ajania. There is extensive evolutionary rate heterogeneity in plastid genes. The psbH and ycf2 genes, which are involved in photosynthesis and ATP transport, are under selective pressure. Plastomes from Ajania species diverged, and structural aspects of plastomes may indicate some of the real evolutionary connections. We suggest the ycf1 gene as a viable plastid DNA barcode because it has significant nucleotide diversity and better reflects evolutionary connections. CONCLUSION: Our findings validate the early Ajania taxonomy reclassification and show evolutionary rate heterogeneity, genetic variety, and phylogenetic heterogeneity of plastid genes. This research might provide new insights into the taxonomy and evolution of Ajania, as well as provide useful information for germplasm innovation and genetic enhancement in horticultural species.

[Research progress on chemical constituents and pharmacological effects of Ajania plants].

Ajania belonging to the subtribe Artemisiinae of Anthemideae(Asteraceae) is a genus of semi-shrubs closely related to Chrysanthemum. There are 24 species of Ajania in northwestern China, most of which are folk herbal medicines with strong stress tolerance. Modern medical studies have demonstrated that the chemical constituents of Ajania mainly include terpenoids, flavonoids, phenylpropanoids, alkynes, and essential oils. These compounds endow the plants with antimicrobial, anti-inflammatory, antitumor, antimalarial, antioxidant, and insecticide effects. In this study, we reviewed the research progress in the chemical constituents and pharmacological activities of Ajania, aiming to provide reference for the further research and development of Ajania.

Research progress on chemical constituents and pharmacological effects of Ajania plants / 中国中药杂志

Ajania belonging to the subtribe Artemisiinae of Anthemideae(Asteraceae) is a genus of semi-shrubs closely related to Chrysanthemum. There are 24 species of Ajania in northwestern China, most of which are folk herbal medicines with strong stress tolerance. Modern medical studies have demonstrated that the chemical constituents of Ajania mainly include terpenoids, flavonoids, phenylpropanoids, alkynes, and essential oils. These compounds endow the plants with antimicrobial, anti-inflammatory, antitumor, antimalarial, antioxidant, and insecticide effects. In this study, we reviewed the research progress in the chemical constituents and pharmacological activities of Ajania, aiming to provide reference for the further research and development of Ajania.

Simultaneous determination of five constituents in Artemisia sieversiana and Ajania tenuifolia by HPLC / 中成药

AIM To establish an UPLC-MS/MS method for the simultaneous content determination of chlorogenic acid,umbellifolactone,rutin,isochlorogenic acid A and isochlorogenic acid C in Artemisia sieversiana Ehrhart ex Willd.and Ajania tenuifolia(Jacquem.ex DC.)Tzvelev in Schischk.&Bobrov by HPLC,and compare the differences between them.METHODS The analysis of 50%methanol extract of both was performed on a 30℃thermostatic CAPCELL PAK-C18 column(100 mm×2.1 mm,1.7 μm),with the mobile phase comprising of acetonitrile-0.2%phosphoric acid water,flowing at 1 mL/min in a gradient elution manner,and the detection wavelengths were set at 290 nm.A t-test was used to compare the contents differences of five constituents in the two crude drugs.RESULTS Five constituents showed good linear relationships within their own ranges(r≥0.999 5),whose average recoveries were 96.03%-103.11%,with the RSDs of 0.51%-2.26%.The average contents of chlorogenic acid,umbellifolactone,rutin,isochlorogenic acid A and isochlorogenic acid C in A.sieversiana and A.tenuifolia were 3.61,0.71,1.46,8.74,1.44 mg/g and 2.60,0.26,0.79,6.31,1.22 mg/g,respectively.CONCLUSION The method is accurate,reliable and reproducible.There was no significant difference in the five chemical constituents between A.sieversiana and A.tenuifolia.

Phytotoxic, insecticidal, and antimicrobial activities of Ajania tibetica essential oil.

The chemical profile of Ajania tibetica essential oil (EO) and its phytotoxic, insecticidal, and antimicrobial activities were assessed. Monoterpenes (79.05%) and sesquiterpenes (10.33%) were dominant in the EO, with camphor, (+/-)-lavandulol and eucalyptol being the major constituents, representing 55.06% of the total EO. The EO possessed potent phytotoxicity against Poa annua and Medicago sativa starting from 0.5 mg/mL, and when the concentration rose to 5 mg/mL, seed germination of both tested species was 100% suppressed. Ajania tibetica EO displayed significant pesticidal activity against Aphis gossypii with an LC50 value of 17.41 µg/mL; meanwhile, the EO also showed antimicrobial activity against Escherichia coli, Bacillus subtilis, Verticillium dahlia and Aspergillus niger using broth microdilution and disc diffusion methods. For the tested bacterial and fungal strains, the EO exhibited a repressing effect, with minimum inhibitory concentrations (MICs) ranging from 0.3125 to 1.25 mg/mL for bacteria and from 1.25 to 2.5 mg/mL for fungi, whereas the minimum microbicidal concentrations (MMCs) were 5 mg/mL for bacteria and 2.5 mg/mL for fungi. Our study is the first report on the chemical profile as well as the phytotoxicity, insecticidal and antimicrobic activity of A. tibetica EO, indicating its potential value as an alternative synthetic pesticide.

Ajania purpurea Extract Attenuates LPS-Induced Inflammation in RAW264.7 Cells and Peritonitis Mice.

Macrophages have important roles in the progression of inflammation. Ajania purpurea Shih. is a member of the Ajania Poljakor family that grows in Tibet (China). Extracts from plants in this genus have anti-bacterial and anti-inflammatory properties. However, there are few reports on the activity and mechanism of Ajania purpurea. Here, we confirmed the anti-inflammatory effect of Ajania purpurea Shih. ethanol extract (EAPS) by examining the levels of inflammatory factors in a mouse model of peritonitis and RAW264.7 cells. The main components of EAPS detected by LC-MS analysis included piperine and chlorogenic acid. In particular, in lipopolysaccharide (LPS)-induced RAW264.7 cells, EAPS inhibited the protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in LPS-induced RAW264.7 cells, lowered the levels of nitric oxide (NO) and prostaglandin E2 (PGE2), as well as the release of inflammatory factors such as tumor necrosis factor-alpha (TNF-α) and pro-inflammatory cytokines such as interleukin (IL)-1ß and IL-6. In addition, Western blot analysis and immunofluorescence staining verified that EAPS inhibited the activity of the nuclear factor-kappaB (NF-κB) pathway by reducing the nuclear translocation of the p65 subunit. Furthermore, in a mouse model of peritonitis, EAPS inhibited the release of inflammatory factors, as well as the recruitment of immune cells including neutrophils and macrophages. These findings indicated that EAPS suppressed LPS-induced inflammation via inhibiting the NF-κB pathway in RAW264.7 cells and mice with peritonitis. Thus, EAPS may be a viable therapeutic method for the treatment of inflammation and related disorders.

The complete sequence of the chloroplast genome of Chrysanthemum rupestre, a diploid disciform capitula species of Chrysanthemum.

In this study, we analyzed the complete sequence of the chloroplast genome of Chrysanthemum rupestre Matsum. et Koidz., 1910, a diploid disciform capitula species of Chrysanthemum endemic to Japan, formerly classified as Ajania rupestris (Matsum. & Koidz.) Muldashev, Bot. Zhurn. (Moscow & Leningrad), 1983. The chloroplast genome of C. rupestre has a typical conserved quadripartite structure of 151,061 bp in length, comprising a large single-copy region (82,846 bp), a small single-copy region (18,301 bp), and a pair of inverted repeat regions (each 24,957 bp). Phylogenetic analysis indicated that C. rupestre clustered with other Chrysanthemum species, including another former Ajania species, Chrysanthemum pacificum Nakai, 1928. However, Ajania variifolia (C.C.Chang) Tzvelev, 1961, which is a synonym of Phaeostigma variifolium (C.C.Chang) Muldashev, 1981, was placed outside the Chrysanthemum clade, thereby implying that the former genus Ajania includes heterogeneous species.

Clarifying Recent Adaptive Diversification of the Chrysanthemum-Group on the Basis of an Updated Multilocus Phylogeny of Subtribe Artemisiinae (Asteraceae: Anthemideae).

Understanding the roles played by geography and ecology in driving species diversification and in the maintenance of species cohesion is the central objective of evolutionary and ecological studies. The multi-phased orogenesis of Qinghai-Tibetan Plateau (QTP) and global climate changes over late-Miocene has profoundly influenced the environments and evolution of organisms in this region and the vast areas of Asia. In this study, we investigate the lineage diversification of Chrysanthemum-group in subtribe Artemisiinae (tribe Anthemideae, Asteraceae) likely under the effects of climate changes during this period. Using DNA sequences of seven low-copy nuclear loci and nrITS and the coalescent analytical methods, a time-calibrated phylogeny of subtribe Artemisiinae was reconstructed with emphasis on Chrysanthemum-group. The monophyletic Chrysanthemum-group was well resolved into two major clades corresponding to Chrysanthemum and Ajania, two genera which can be well identified by capitulum morphology but have been intermingled in previous plastid and ITS trees. Within Chrysanthemum, a later divergence between Ch. indicum-complex and Ch. zawadskii-complex can be recognized. The time frames of these sequential divergences coincide with the late Cenozoic uplift of the Northern QTP and the concomitant climatic heterogeneity between eastern and inland Asia. Reconstruction of historical biogeography suggested the origin of Chrysanthemum-group in Central Asia, followed by eastward migration of Chrysanthemum and in situ diversification of Ajania. Within Chrysanthemum, Ch. indicum-complex and Ch. zawadskii-complex exhibited contemporary distributional division, the former in more southern and the latter in more northern regions. The geographic structure of the three lineages in Chrysanthemum-group have been associated with the niche differentiation, and environmental heterogenization in Asia interior.

Dysfunction of CYC2g is responsible for the evolutionary shift from radiate to disciform flowerheads in the Chrysanthemum group (Asteraceae: Anthemideae).

Evolutionary shifts among radiate, disciform and discoid flowerheads have occurred repeatedly in a number of major lineages across the Asteraceae phylogeny; such transitions may also appear within evolutionarily young groups. Although several studies have demonstrated that CYC2 genes partake in regulating floral morphogenesis in Asteraceae, the evolution of capitulum forms within a recently diverging lineage has remained poorly understood. Here, we study the molecular regulation of the shift from a radiate to a disciform capitulum within the Chrysanthemum group. This is a recently radiating group mainly comprising two genera, Chrysanthemum and Ajania, that are phylogenetically intermingled but distinct in flowerhead morphology: Chrysanthemum spp. with radiate capitula and Ajania spp. with disciform capitula. We found that the morphogenesis of zygomorphy in the marginal floret in Ajania was disrupted soon after floral primordium emergence; CYC2g, one of the CYC2 copies that was expressed prominently in the ray floret of Chrysanthemum was not expressed in flowerheads of Ajania. Weakening the expression of ClCYC2g in Chrysanthemum lavandulifolium led to the gradual transition of a ray flower toward the disc-like form. Molecular evolutionary analyses indicated that the disciform capitulum might have evolved only once, approximately 8 Mya, arising from dysfunction of the CYC2g orthologs. A 20-nt deletion, including a putative TATA-box of the Ajania-type CYC2g promoter, appeared to inhibit the expression of the gene. Considering the divergent habitats of Chrysanthemum and Ajania, we propose that the shift from radiate to disciform capitulum must have been related to changes in pollination strategies under selective pressure.

The complete chloroplast genome sequence of Ajania pacifica (Nakai) Bremer & Humphries.

The complete chloroplast (cp) genome sequence of Ajania pacifica, called as golden and silver chrysanthemum, was newly analyzed in this study. It was 151,059 bp in length and was a typical circular structure composed of and comprised of a large single copy region (82,857 bp) and a small single copy region (18,294 bp) which were separated by two inverted repeat regions (24,954 bp). The molecular phylogenetic analyses of A. pacifica and its related taxa was conducted based on the complete chloroplast genome sequences, and it was proved that the genus Ajania is embedded in the genus Chrysanthemum clade as well as a monotypic genus Opisthopappus. In the other hand, the genus Artemisia was divided into two group in the tribe Anthemideae.

Insecticidal Activities and Chemical Composition of the Essential Oils of Ajania nitida and Ajania nematoloba from China.

In this work, we investigated insecticidal and repellent activities of the essential oils extracted from Ajania nitida and Ajania nematoloba against Tribolium castaneum and Lasioderma serricorne adults. The components of essential oils were analyzed by GC-MS. The main components of A. nitida oil were camphor (20.76%), thujone (18.64%), eucalyptol (13.42%), borneol (8.32%) and those of A. nematoloba oil were ß-pinene (34.72%), eucalyptol (24.97%) and verbenol (20.39%). The results showed that the two essential oils possessed insecticidal and repellent activities against two species of insects. A. nitida oil possessed contact and fumigant toxicity against T. castaneum (LD50 = 30.10 µg/adult and LC50 = 21.07 mg/L air) and L.serricorne (LD50 = 17.51 µg/adult and LC50 = 11.23 mg/L air). A.nematoloba oil showed contact and fumigant toxicity against T. castaneum (LD50 = 102.29µg/adult and LC50 = 69.45 mg/L air) and contact toxicity against L.serricorne (LD50 = 53.43 µg/adult), but no obvious fumigant effect was observed against L.serricorne. Both of essential oils possessed strong repellent activity against T. castaneum and certain repellent activity against L.serricorne. Especially, A. nematoloba oil showed the same level percentage repellency as DEET(the positve control) against T. castaneum. The results indicated that the essential oils of A. nitida and A. nematoloba had the potential to be developed as natural insecticides and repellents for the control of T. castaneum and L.serricorne.

Chemical Constituents and Insecticidal Activities of Ajania fruticulosa Essential Oil.

The insecticidal activity and chemical constituents of the essential oil from Ajania fruticulosa were investigated. Twelve constituents representing 91.0% of the essential oil were identified, and the main constituents were 1,8-cineole (41.40%), (+)-camphor (32.10%), and myrtenol (8.15%). The essential oil exhibited contact toxicity against Tribolium castaneum and Liposcelis bostrychophila adults with LD50 values of 105.67 µg/adult and 89.85 µg/cm(2) , respectively. The essential oil also showed fumigant toxicity against two species of insect with LC50 values of 11.52 and 0.65 mg/l, respectively. 1,8-Cineole exhibited excellent fumigant toxicity (LC50  = 5.47 mg/l) against T. castaneum. (+)-Camphor showed obvious fumigant toxicity (LC50  = 0.43 mg/l) against L. bostrychophila. Myrtenol showed contact toxicity (LD50  = 29.40 µg/cm(2) ) and fumigant toxicity (LC50  = 0.50 mg/l) against L. bostrychophila. 1,8-Cineole and (+)-camphor showed strong insecticidal activity to some important insects, and they are main constituents of A. fruticulosa essential oil. The two compounds may be related to insecticidal activity of A. fruticulosa essential oil against T. castaneum and L. bostrychophila.

Quinones and coumarins from Ajania salicifolia and their radical scavenging and cytotoxic activity.

1,4-Naphthoquinone (1) and a new coumarin (3) were isolated from Ajania salicifolia, together with two known compounds (2, 4). The structures and stereochemistry of new compounds were elucidated using spectroscopic methods. Two compounds exhibited potent ABTS cation radical scavenging activities with IC50 values ranging 7.97-8.44 µM. Two quinones (1, 2) exhibited moderate cytotoxic activity against the human cancer cell lines (Hela, HepG2, and K562) with IC50 values of 11.24-35.15 µM in vitro. This is the first report of naphthoquinone in the genus Ajania.