Benzylpiperazinyl derivatives of alepterolic acid: Synthesis and cytotoxic evaluation.

Natural products play a significant role in the development of modern drugs. Alepterolic acid, a labdane-type diterpenoid firstly isolated from Aleuritopteris argentea (Gmél.) Fée, has been identified as a valuable template for the synthesis of potent anticancer agents by structural modification. In this study, a series of new derivatives was obtained by coupling alepterolic acid with benzylpiperazines. It was found that (3,4-dichlorobenzyl)piperazinyl alepterolic acid (compound 6p) displayed the most toxic against MCF-7 cell line, with IC50 value of 8.31±0.67 µM. Further investigations demonstrated that compound 6p induced morphological changes in MCF-7 cells, inhibited proliferation in a time- and dose-dependent manner. Furthermore, western blot analysis revealed that compound 6p induced a significant increase in cleaved caspase-9, cleaved caspase-3, cleaved poly (ADP-ribose) polymerase (PARP) and Bax/Bcl2 ratio in MCF-7 cells. All of these results confirmed that compound 6p induced endogenous apoptosis in MCF-7 cells. Conclusively, the findings suggest that the incorporation of benzylpiperazine to alepterolic acid represents a promising approach for the discovery of new drug candidates.

New Chlorine-Containing Sesquiterpenoid from Artemisia Blepharolepis.

One new chlorinated sesquiterpenoid (compound 1, ablepharolide) and twenty-one known compounds were obtained from the aerial parts of Artemisia blepharolepis. Their structures were established by spectroscopic methods and the absolute configuration was further determined by single-crystal X-ray diffraction analysis for ablepharolide. Ablepharolide is a rare sesquiterpenoid with a 4-methyl-7-isopropyl-9-ethyl-perhydroindene skeleton that incorporates a chlorine atom. It significantly inhibited the growth of MCF-7 cells with IC50 value of 8.34±0.77 µM. Further investigations demonstrated that ablepharolide induced morphological changes in MCF-7 cells, inhibited proliferation in a time- and dose-dependent manner. Furthermore, western blot analysis revealed that ablepharolide induced a significant increase in cleaved caspase-8, cleaved caspase-3 and cleaved poly(ADP-ribose) polymerase (PARP) in MCF-7 cells. All of these results supported that ablepharolide induced exogenous apoptosis in MCF-7 cells.

Reliability and validity of the Chinese version of the selective control assessment of the lower extremity in children with spastic cerebral palsy.

Objective: To assess the reliability and validity of the Chinese version of the Selective Control Assessment of the Lower Extremity (SCALE) in children with spastic cerebral palsy (CP). Methods: Forty-five children with spastic CP (mean age 7.29 years, SD 2.87 years, rang 4-16 years) were recruited. Internal consistency was measured using Cronbach's α, while test-retest and inter-rater reliability were evaluated using intra-class correlation coefficients (ICC). Construct validity was established through correlation and confirmatory factor analyses. Discriminative validity was assessed by comparing SCALE scores across varying GMFCS levels. Results: The Chinese version of SCALE demonstrates high internal consistency (Cronbach's α = 0.91) and good reliability with ICCs exceeding 0.76 for test-retest and inter-rater assessments. It shows significant correlations with GMFCS (r = -0.76, p < 0.001) and Fugl-Meyer scales (r = 0.79, p < 0.001), confirming its validity. Confirmatory factor analysis supports a well-fitting model (χ 2/df = 1.58, RMSEA = 0.08, SRMR <0.001, GFI = 0.98, AGFI = 0.90, CFI = 0.99, TLI = 0.98), with the latent variable's AVE at 0.59 and CR at 0.88. Discriminative validity is evident in significant differences across GMFCS levels (p < 0.001), notably between levels I and II, I and III, and I and IV (p < 0.05). Conclusion: The Chinese version of SCALE shows good reliability and validity for assessing lower limb selective movement control in children with spastic cerebral palsy in China. Clinical trial registration: https://www.chictr.org.cn/showproj.html?proj=205380, identifier ChiCTR2400083880.

Antibacterial effect of Cu2O/TiO2 visible-light photocatalytic composite on Xanthomonas campestris.

In this study, a Cu2O/TiO2 (CuTi) visible-light photocatalytic composite was employed for the treatment of Xanthomonas campestris and X. campestris-infected Brassica napus seedlings. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values against X. campestris were determined to be 8 and 32 µg ml-1, respectively. Transmission electron microscopy analysis demonstrated a direct correlation between the extent of bacterial cell damage and the concentration of CuTi. Noteworthily, a bactericidal rate of 100% was achieved at a concentration of 150 µg ml-1 over a treatment duration of 120 min. Moreover, alterations in active oxidants and antioxidants, including reactive oxygen species, glutathione reductase, superoxide dismutase, peroxidase, and catalase within the bacterial cells, were examined to elucidate the underlying mechanism of inhibition by the CuTi. The B. napus infected by X. campestris was treated with CuTi, and the efficacy was validated through determination of plant resistance indexes. The combined data confirmed that the CuTi is characterized by a low dose, fast onset, good effect, and higher safety for killing X. campestris, and it is expected to be developed as an antimicrobial agent for vegetables.

Case Report: New presentation of CLIFAHDD syndrome with a novel variant in the NALCN gene and a literature review.

Background: Congenital contractures of the limbs and face, hypotonia, and developmental delay (CLIFAHDD) syndrome (OMIM #616266) is an autosomal dominant hereditary disease that can lead to the congenital contracture of the limbs and face, hypotonia, and developmental delay. In addition, it may result in growth retardation and present various clinical symptoms, such as brain atrophy, a small pituitary gland, musculoskeletal abnormalities, abnormal breathing, abdominal hernia, and abnormal facial features. Herein, we describe a novel de novo missense genetic variant in the sodium leak channel, non-selective (NALCN) gene that is associated with CLIFAHDD syndrome. Case description: This study describes a patient with varus deformities in both feet, deviation of the ulnar side of the fingers, and severe hypotonia. This patient was subsequently confirmed to have CLIFAHDD syndrome through genetic testing, which also revealed a novel missense de novo genetic variant in the NALCN gene (c.3553G > A, p.Ala1185Thr). Conclusions: Our findings further enrich the known variant spectrum of the NALCN gene and may expand the range of clinical options for treating NALCN-related disorders.

ent-8(14),15-Pimaradiene-2ß,19-diol, a diterpene from Aleuritopteris albofusca, inhibits growth and induces protective autophagy in hepatocellular carcinoma cells.

A new pimarane-type diterpene, ent-8(14),15-pimaradiene-2ß,19-diol (JXE-23), was isolated from the fern plant Aleuritopteris albofusca by our previous work; however, the biological activity of this diterpene remains unclear. In the present study, the anti-cancer potential of JXE-23 in various cancer cells was investigated. Among MCF-7 breast cancer cells, A549 lung cancer cells, and HepG2 liver cancer cells, JXE-23 displayed significant cytotoxicity to HepG2 cells with an IC50 value of 17.20 ± 1.73 µM, while showing no obvious toxicity in normal hepatocytes HL7702. JXE-23 inhibited cell growth and colony formation in HepG2 cells. A cell cycle distribution analysis showed that JXE-23 caused G2/M cell cycle arrest. Besides, JXE-23 also suppressed the migration of HepG2 cells. Interestingly, an increase of light chain 3 II (LC3II) and Beclin 1 and a decrease of P62 have occurred in JXE-23-treated cells, as well as the formation of GFP-LC3 dots, indicative of autophagy induction by JXE-23. When combined with autophagy inhibitor 3-methyladenine and chloroquine, the cell viability was significantly reduced, suggesting that JXE-23 triggered protective autophagy in hepatoma cells. Further study showed that JXE-23 inactivated the CIP2A/p-AKT/c-Myc signaling axis in HepG2 cells. Our data provided evidence that JXE-23 inhibited cell growth, arrested cells at the G2/M phase, and induced protective autophagy in HepG2 hepatocellular carcinoma cells. JXE-23 may be a potential lead compound for anti-cancer drug development, and autophagy inhibitor treatment may provide an effective strategy for improving its anti-cancer effect.

Discovery of a Pimaradiene that Decreases Viability of MDA-MB-468 Cells Through Inhibition of EGFR Signaling Pathway.

Triple-negative breast cancer (TNBC) is characterized by strong invasiveness, high relapse rates, and poor overall survival. It occurs in approximately 15-20 % of all breast cancer cases. Natural compounds are a promising option for managing breast cancer. ent-8(14),15-Pimaradiene-2ß,19-diol (JXE-23), is a pimaradiene isolated from the fern Aleuritopteris albofusca. However, the effects and molecular mechanisms of JXE-23 on cancer cells are still unknown. Thus, this study was designed to determine the potential of JXE-23 for its anticancer properties in TNBC cells. JXE-23 was evaluated for its antiproliferative activity in vitro against human breast cancer cell lines, and showed selectively cytotoxic activity against MDA-MB-468, an EGFR-overexpressing TNBC cancer cell line, with an IC50 value of 1.17±0.04 µM. Moreover, mechanistic investigations indicated that JXE-23 was significantly capable of inhibiting cell proliferation and viability in MDA-MB-468 cells. In addition, JXE-23 exerted an anticancer effect against MDA-MB-468 cells via restraining cell migration in a dose-dependent mode. Moreover, after treatment with JXE-23, the protein expressions of pEGFR, pERK, pAkt and p-p70S6K were significantly reduced in MDA-MB-468 cells. The results underscored that JXE-23 could be a potential lead compound for the treatment of EGFR-overexpressing TNBC cells.

Anti-inflammatory activities of several diterpenoids isolated from Hemionitis albofusca.

Hemionitis albofusca (Baker) Christenh is a plant that grows in various regions of China. Although it is not recognized as a traditional medicine, it is often mistakenly labelled and used as Aleuritopteris argentea (S. G. Gmél.) Fée to alleviate menstruation-related issues. Recently, several diterpenoids such as ent-16-oxo-17-norkauran-19-oic acid (Compound A), 14-oxy-7ß,20-dihydroxycyath-12,18-diene (Compound B), ent-8(14),15-pimaradiene-2ß,19-diol (Compound C), ent-kaurane-16-ene-2ß,18α-diol (Compound D), ent-kaurane-2ß,16α,18α-triol (Compound E), and onychiol B have been extracted from H. albofusca. In this study, we investigated the anti-inflammatory activity of these diterpenes. We confirmed that compounds A ~ D suppressed the amount of cellular NO production by inhibiting the expression and transcription of iNOS protein. They also significantly inhibited the expression and transcription of inflammatory factors TNF-α and IL-6. Additionally, Compounds A and C suppressed the activation of the NF-κB signaling pathway and inhibited the phosphorylation level of p38, ultimately down-regulating inflammation. Compound B suppressed the activation of the NF-κB signaling pathway, while Compound D inhibited the phosphorylation level of p38 and down-regulated the activation of the p38 MAPK signaling pathway. In a word, our investigation supports the potential application of natural diterpenes as lead compounds for developing anti-inflammatory agents.

Identification of an Alepterolic Acid Derivative as a Potent Anti-Breast-Cancer Agent via Inhibition of the Akt/p70S6K Signaling Pathway.

Alepterolic acid is a diterpene occurring in the fern Aleuritopteris argentea with potential biological activity that warrants further structural modification. In the present work, sixteen alepterolic acid derivatives were synthesized and evaluated for their anticancer activities. Among them, N-[m-(trifluoromethoxy)phenyl] alepterolamide displayed comparable activity (IC50 =4.20±0.21 µM) in MCF-7 cells. Moreover, mechanistic investigations indicated this compound was significantly capable of diminishing cell proliferation and viability of MCF-7 cells. After treatment with N-[m-(trifluoromethoxy)phenyl] alepterolamide, a significant increase in cleaved caspase-9, cleaved caspase-3, cleaved poly (ADP-ribose) polymerase (PARP) and Bax/Bcl2 ratio were observed in MCF-7 cells, leading to caspase-dependent apoptotic pathways. Further studies showed this compound promoted cellular apoptosis and inhibited migration in MCF-7 cells via modulation of the Akt/p70S6K signaling pathway. All these results revealed the potential of N-[m-(trifluoromethoxy)phenyl] alepterolamide as an appealing therapeutic drug candidate for breast cancer.

Stepwise changes in flavonoids in spores/pollen contributed to terrestrial adaptation of plants.

Protecting haploid pollen and spores against UV-B light and high temperature, 2 major stresses inherent to the terrestrial environment, is critical for plant reproduction and dispersal. Here, we show flavonoids play an indispensable role in this process. First, we identified the flavanone naringenin, which serves to defend against UV-B damage, in the sporopollenin wall of all vascular plants tested. Second, we found that flavonols are present in the spore/pollen protoplasm of all euphyllophyte plants tested and that these flavonols scavenge reactive oxygen species to protect against environmental stresses, particularly heat. Genetic and biochemical analyses showed that these flavonoids are sequentially synthesized in both the tapetum and microspores during pollen ontogeny in Arabidopsis (Arabidopsis thaliana). We show that stepwise increases in the complexity of flavonoids in spores/pollen during plant evolution mirror their progressive adaptation to terrestrial environments. The close relationship between flavonoid complexity and phylogeny and its strong association with pollen survival phenotypes suggest that flavonoids played a central role in the progression of plants from aquatic environments into progressively dry land habitats.

Antitumor Cembrane Diterpenoids from the South China Sea Soft Coral Lobophytum sp.

Two new highly functionalized cembrane diterpenoids named ximaolobophytolides A (1) and B (2) as minor components, together with seven related known compounds (3-9), have been isolated and identified from the Ximao soft coral Lobophytum sp. They were characterized by the presence of an α-methylene-γ-lactone moiety. Based on the comprehensive analyses of 1D and 2D NMR spectroscopic data, the absolute configurations of these two new compounds were elucidated by the combination of quantum mechanical (QM)-NMR and time-dependent density functional theory/electronic circular dichroism (TDDFT-ECD) calculation approaches. In the anti-tumor bioassays, compounds 3-9 showed moderate to significant inhibitory effects (IC50 values ranging from 29.66 to 0.39 µM) against the proliferations of five tumor cells HEL, A549, H1975, MDA-MB-231, and H1299. It might be worthy to point out that compounds 4, 7, and 8 exhibited better anti-tumor activities than that of the positive control Doxorubicin.

Synthesis and Evaluation of Piperazine-Tethered Derivatives of Alepterolic Acid as Anticancer Agents.

Alepterolic acid is a natural diterpenoid isolated from Aleuritopteris argentea with potential anti-cancer activity. In this study, alepterolic acid was modified to construct a series of arylformyl piperazinyl derivatives (3a-3p). The synthesized derivatives were fully characterized with HRMS, NMR, and IR. Four compounds with inhibition rate higher than 30 % at 10 µM (3f, 3n, 3g and 3k) were further measured to obtain the IC50 values against four cancer cell lines, including hepatoma cell lines HepG2, lung cancer cell lines A549, estrogen receptor-positive cell lines MCF7, and triple-negative breast cancer (TNBC) cell lines MDA-MB-231 by MTT assay. It was found that these compounds were more effective to HepG2 and MDA-MB-231 cells, while less toxic to A549 and MCF7 cells, and compound 3n as the most toxic derivatve against MDA-MB-231 cell lines, with IC50 value of 5.55±0.56 µM. Trypan blue staining and colony formation assay showed that compound 3n inhibited the growth of MDA-MB-231 cells and prevented colony formation. Hoechst staining, flow cytometry and western blot analysis revealed that compound 3n induced caspase-dependent apoptosis in MDA-MB-231 cells. Conclusively, compound 3n was demonstrated to be a potential anti-cancer lead compound for further investigation.

Structurally Diverse Diterpenes from the South China Sea Soft Coral Sarcophyton trocheliophorum.

The present investigation of the South China Sea soft coral Sarcophyton trocheliophorum resulted in the discovery of six new polyoxygenated diterpenes, namely sartrocheliols A-E (1, 3, 5-8) along with four known ones, 2, 4, 9, and 10. Based on extensive spectroscopic data analysis, sartrocheliol A (1) was identified as an uncommon capnosane diterpene, while sartrocheliols B-E (3, 5-8) were established as cembrane diterpenes. They displayed diverse structural features not only at the distinctly different carbon frameworks but also at the various types of heterocycles, including the epoxide, γ-lactone, furan, and pyran rings. Moreover, their absolute configurations were determined by a combination of quantum mechanical-nuclear magnetic resonance (QM-NMR) approach, modified Mosher's method, and X-ray diffraction analysis. In the anti-tumor bioassay, compound 4 exhibited moderate cytotoxic activities against A549, H1975, MDA-MB-231, and H1299 cells with the IC50 values ranging from 26.3 to 47.9 µM.

Elucidating Flavonoid and Antioxidant Activity in Edible and Medicinal Herbs Woodwardia japonica (L.f.) Sm. Based on HPLC-ESI-TOF-MS and Artificial Neural Network Model: Response to Climatic Factors.

Woodwardia japonica is a kind of great potential edible and medicinal fern. In a previous study, it was found that flavonoid and antioxidant activity of W. japonica from different sites were different. However, the cause of the differences has still been unclear, which has restricted the utilization of W. japonica. In this paper, flavonoid and antioxidant activity of W. japonica from nine different regions were determined with the method of a colorimetric assay with UV-VIS spectrophotometry and HPLC-ESI-TOF-MS, and the effects of climate factors on flavonoids and antioxidant activities were evaluated by mathematical modeling and statistical methods. The results showed: (1) total flavonoid content (TFC) of W. japonica from Wuyi Mountain (Jiangxi) was the highest, which might be related to the low temperature; (2) the differences of antioxidant activities of W. japonica might be related to precipitation; (3) five flavonols, two flavones and one isoflavone were tentatively identified in W. japonica; (4) flavonol and isoflavone might be affected by sunshine duration, and flavones were probably related to temperature. In conclusion, the effects of climate factors on flavonoids and antioxidants are significant, which would provide an important basis for further exploring the mechanism of climate affecting secondary metabolites.

KCTD10 functions as a tumor suppressor in hepatocellular carcinoma by triggering the Notch signaling pathway.

OBJECTIVE: Our previous study found KCTD10 negatively regulates Notch signaling, but whether KCTD10 regulates human hepatocellular carcinoma (HCC) carcinogenicity was uncertain. METHODS: We used lentivirus infection to regulate KCTD10 expression in HCC cell lines, then monitored tumor sphere formation rate, cell migration, in vitro and in vivo tumorigenicity, cancer stem cell (CSC) biomarkers and Notch signaling variation. RESULTS: Down-regulation of KCTD10 in HCC cell lines (Hep3B and MHCC97H) enhanced the expression of CSC marker genes, promoted self-renewal and tumorigenic ability, and increased the CD133+ cell population. Further molecular studies showed that the transmembrane/intracellular region (NTM) of Notch1 decreased when KCTD10 was knocked down in HCC cell lines, and that the balance between P53 and Notch activity was regulated. CONCLUSIONS: The results demonstrated that KCTD10 can act as a tumor suppressor in HCC cells through Notch signaling.

Commentary: Machine learning for autism spectrum disorder diagnosis - challenges and opportunities - a commentary on Schulte-Rüther et al. (2022).

The commentary cites a study by Schulte-Rüther et al. (Journal of Child Psychology and Psychiatry, 2022) that proposed a machine learning model to predict a clinical best-estimate diagnosis of ASD when existing other co-occurring diagnoses. We discuss the valuable contribution of this work to developing a reliable computer-assisted diagnosis (CAD) system for ASD and point out that related research can be integrated with other multimodal machine learning methods. For future studies on developing the CAD system for ASD, we propose problems that need to be solved and potential research directions.

Onychiol B attenuates lipopolysaccharide-induced inflammation via MAPK/NF-κB pathways and acute lung injury in vivo.

Acute lung injury (ALI) is a devastating respiratory disorder characterized by rapid alveolar injury, uncontrolled inflammatory response, etc. Onychiol B is a cyathane diterpene originally isolated from fern plants. In this study, onychiol B can inhibit the production and secretion of pro-inflammatory cytokines such as NO, iNOS, IL-6 and TNF-α in LPS-stimulated RAW264.7 cells by restraining the NF-κB and the p38 MAPK pathway. In addition, it prevents the production of ROS and reduces the loss of mitochondrial membrane potential in LPS-stimulated RAW264.7 cells. Furthermore, in the acute lung injury mouse model induced by LPS injected into the trachea, onychiol B alleviates pulmonary edema, reverses inflammatory mediator TNF-α, IL-6, and IL-ß secretion in lung. In general, our data show that significant anti-ALI effects of onychiol B would render it a potential candidate for the treatment of inflammatory diseases.

Casticin Attenuates Stemness in Cervical Cancer Stem-Like Cells by Regulating Activity and Expression of DNMT1.

OBJECTIVE: To explore whether casticin (CAS) suppresses stemness in cancer stem-like cells (CSLCs) obtained from human cervical cancer (CCSLCs) and the underlying mechanism. METHODS: Spheres from HeLa and CaSki cells were used as CCSLCs. DNA methyltransferase 1 (DNMT1) activity and mRNA levels, self-renewal capability (Nanog and Sox2), and cancer stem cell markers (CD133 and CD44), were detected by a colorimetric DNMT activity/inhibition assay kit, quantitative real-time reverse transcription-polymerase chain reaction, sphere and colony formation assays, and immunoblot, respectively. Knockdown and overexpression of DNMT1 by transfection with shRNA and cDNA, respectively, were performed to explore the mechanism for action of CAS (0, 10, 30, and 100 nmol/L). RESULTS: DNMT1 activity was increased in CCSLCs compared with HeLa and CaSki cells (P<0.05). In addition, HeLa-derived CCSLCs transfected with DNMT1 shRNA showed reduced sphere and colony formation abilities, and lower CD133, CD44, Nanog and Sox2 protein expressions (P<0.05). Conversely, overexpression of DNMT1 in HeLa cells exhibited the oppositive effects. Furthermore, CAS significantly reduced DNMT1 activity and transcription levels as well as stemness in HeLa-derived CCSLCs (P<0.05). Interestingly, DNMT1 knockdown enhanced the inhibitory effect of CAS on stemness. As expected, DNMT1 overexpression reversed the inhibitory effect of CAS on stemness in HeLa cells. CONCLUSION: CAS effectively inhibits stemness in CCSLCs through suppression of DNMT1 activation, suggesting that CAS acts as a promising preventive and therapeutic candidate in cervical cancer.