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Abstract Endoplasmic reticulum (ER) stress can trigger various cell death mechanisms beyond apoptosis, providing promise in cancer treatment. Oncosis, characterized by cellular swelling and increased membrane permeability, represents a non-apoptotic form of cell death. In our study, we discovered that Arnicolide D (AD), a natural sesquiterpene lactone compound, induces ER stress-mediated oncosis in hepatocellular carcinoma (HCC) cells, and this process is reactive oxygen species (ROS)-dependent. Furthermore, we identified the activation of the PERK-eIF2α-ATF4-CHOP pathway during ER stress as a pivotal factor in AD-induced oncosis. Notably, the protein synthesis inhibitor cycloheximide (CHX) was found to effectively reverse AD-induced oncosis, suggesting ATF4 and CHOP may hold crucial roles in the induction of oncosis by AD. These proteins play a vital part in promoting protein synthesis during ER stress, ultimately leading to cell death. Subsequent studies, in where we individually or simultaneously knocked down ATF4 and CHOP in HCC cells, provided further confirmation of their indispensable roles in AD-induced oncosis. Moreover, additional animal experiments not only substantiated AD’s ability to inhibit HCC tumor growth but also solidified the essential role of ER stress-mediated and ROS-dependent oncosis in AD’s therapeutic potential. In summary, our research findings strongly indicate that AD holds promise as a therapeutic agent for HCC by its ability to induce oncosis.

Objective: To evaluate the synergistic effects and underlying mechanisms of WNT974 and artesunate (ART) combination therapy in the treatment of colorectal cancer (CRC). Methods: We conducted an initial assessment of the synergistic impact of WNT974 and ART on cell viability using the MTT assay across eight CRC cell lines with diverse mutation statuses.Subsequently, a 3D in vitro model was utilized to confirm the synergistic effect of the combination. Flow cytometry was employed to analyze cell cycle distribution. The molecular mechanisms were further investigated by examining the expression levels of cell cycle-related proteins (p21, p27, and CDK2) and AKT activity through PCR and Western blot analyses. The in vivo efficacy of the combination therapy was evaluated using a xenograft mouse model of CRC. Results: The combination of WNT974 and ART significantly reduced cell viability in all eight CRC cell lines, demonstrating a synergistic effect. Additionally, the combination exhibited synergy in inhibiting 3D spheroid growth. Cell cycle analysis revealed that the combination therapy induced a marked G1/G0 arrest through CDK2 inhibition, surpassing the efficacy of individual treatments. Molecular analysis indicated that the combination therapy upregulated p21 and p27 expression and reduced AKT activity. In vivo, the combination therapy significantly inhibited tumor growth in the xenograft mouse model, notably outperforming 5FU, the standard first-line drug for CRC. Conclusion: The study highlights the synergistic anti-CRC effects of combining WNT974 and ART. This combination induces G1/G0 phase cell cycle arrest by cooperatively regulating p21, p27, and AKT, presenting a promising alternative to current CRC treatments. These findings elucidate the molecular basis of this interaction and provide a scientific foundation for advancing ART combined with WNT94 as a novel therapy for CRC, offering a potential new avenue.

Comprehending the mechanisms governing metacommunity assembly holds pivotal importance for effective conservation strategies. Amidst diverse theories in explaining metacommunity assembly processes, ecologists are progressively delving into the interplay between ecological stochasticity and determinism in shaping species assemblages. Given the ease of measurement, three null model-based methodologies have been widely employed to measure the stochastic and deterministic process, i.e., dispersal-niche continuum index (DNCI), elements of metacommunity structure (EMS), and zeta diversity calculations. However, it is still in doubt which approach can better detect the dominant ecological process governing metacommunity assembly. In this study, we concurrently utilized these three approaches to reveal the fish metacommunity assembly processes in a transitional floodplain among different habitats between high- and low-water levels. In general, our results recognized functional zeta diversity as indicative measurement in this study case since it uniquely unveiled the significant assembly transitions from stochasticity to determinism in response to water receding. Specifically, fish metacommunity in high-water periods was governed by stochastic processes, which can be ascribed to connectivity and easier dispersal in the flooding period. In contrast, the deterministic process shaped fish metacommunity in low-water phase since the stringent environmental loading enhance environmental filtering. This phenomenon is more pronounced in lotic habitats due to the more intricate environments there. It is worth noting that EMS and DNCI approaches showed blind response to this shift. In consequence, our findings advocate for favoring zeta diversity in resolving stochastic and deterministic contributions in governing metacommunity assembly. In addition, our findings also highlight the superiority of functional diversity over traditional taxonomic diversity, since we recorded a more reasonable metacommunity assembly process revealed by the functional group approach, which emphasizes the significance of incorporating functional traits in community ecology.

Elephantopus scaber, globally acknowledged for its edible and medicinal uses, remains underexplored despite the identification of active compounds. This research explores Elephantopus scaber extract's therapeutic capabilities in hepatocellular carcinoma (HCC). Initial in vitro screenings using the MTT assay revealed that ethanol extract (EEES) inhibited HCC cell proliferation while exhibiting low toxicity to normal cells. Further explorations showed EEES induced cell cycle arrest, apoptosis, and inhibited cell metastasis. A bioinformatics analysis using a network pharmacology approach identified the PI3K/Akt pathway as a key modulation for EEES's anti-HCC activities, validated through western blot analysis. In animal studies, EEES demonstrated the ability to inhibit HCC tumor growth in vivo with a high level of safety. In summary, our research positions Elephantopus scaber as a potent herb with anti-HCC activities and remarkable safety, offering promising prospects for the prevention and treatment of liver diseases, including HCC.

Objective: To evaluate the anticancer activity of the natural diterpenoid lasiokaurin (LAS) against nasopharyngeal carcinoma (NPC) both in vitro and in vivo, along with investigating its underlying mechanism. Methods: MTT and colony formation assays were used to assess cell viability. Flow cytometry was utilized for the analysis of cell cycle arrest and apoptosis. Additionally, wound healing and transwell invasion assays were conducted separately to investigate cell migration and invasion. To uncover potential targets and pathways of LAS within NPC, a network pharmacological study based on RNA-sequencing (RNA-seq) was performed. Immunoblotting was subsequently used to determine protein levels. Furthermore, the anti-NPC activity of LAS was evaluated in vivo using a xenograft mouse model. Results: Based on in vitro investigations, it is evident that LAS exerted substantial inhibitory effects on NPC cells. Notably, LAS demonstrated significant reductions in cell viability, migration, and invasion capabilities, while simultaneously inducing apoptosis and G2/M cell cycle arrest. Furthermore, LAS suppressed the activation of MAPK, mTOR, STAT3, and NF-κB pathways within NPC cells. Additionally, in vivo experiments underscored LAS's capacity to attenuate NPC tumor growth without eliciting any discernible impact on body weight. Conclusion: Our data clearly demonstrate the anticancer activity of LAS against NPC, both in vitro and in vivo. These findings firmly position LAS as a potential candidate for the treatment of NPC.

Stock index is an indicator that describes the changes in the total price level of the stock market, and it is susceptible to many dynamic factors, with such characteristics as high dimension, uncertainty, non-linearity, time delay, complexity, etc., resulting in abnormal and missing values in stock index data, which will lead to instability or unreliability of the stock index tracking model. In order to solve these problems, we take the historical stock index as the input, model the internal dynamic changes of features, and learn the change rule. Firstly, we introduce an attention mechanism, that is, to assign different weights to the implicit state of the long short term memory network (LSTM) through mapping weights and learning parameters. We further propose a stock index data preprocessing model of the LSTM based on the attention mechanism. Secondly, the group method of data handling type neural networks (GMDH-NN) is a self-organizing data mining technology, which is especially suitable for modeling complex systems. So we choose a discrete form of Kolmogorov-Gabor ( $K-G$ ) polynomial of the first-order as the reference function of GMDH-NN to establish the general relationship between input and output variables. We further present a deep evolutionary GMDH polynomial neural network (DGMDH) to perform stock index tracking. Moreover, for a high-dimensional stock index dataset, the traditional external criterion can no longer meet the needs of reality, so we propose a tracking error external criterion (TEEC) for stock indices, which is based on the difference between allocation yield and target yield. The TEEC provides better information for selecting the optimal complex DGMDH model. Our experiments clearly show the effectiveness of our methodology.

Tumor-associated macrophage (TAM) affects the intrinsic properties of tumor cells and the tumor microenvironment (TME), which can stimulate tumor cell proliferation, migration, and genetic instability, and macrophage diversity includes the diversity of tumors with different functional characteristics. Macrophages are now a central drug target in various diseases, especially in the TME, which, as “tumor promoters” and “immunosuppressors”, have different responsibilities during tumor development and accompany by significant dynamic alterations in various subpopulations. Remodelling immunosuppression of TME and promotion of pre-existing antitumor immune responses is critical by altering TAM polarization, which is relevant to the efficacy of immunotherapy, and uncovering the exact mechanism of action of TAMs and identifying their specific targets is vital to optimizing current immunotherapies. Hence, this review aims to reveal the triadic interactions of macrophages with programmed death and oncotherapy, and to integrate certain relationships in cancer treatment.

Abstract Electrochemical water splitting coupled with renewable energy offers a promising avenue for energy conversion and storage, but it is also extremely suppressed by the sluggish kinetics of oxygen evolution reaction (OER) and hydrogen evolution reaction (HER). Herein, a multi‐layer architecture strategy was utilized by the in situ growth for cobalt phosphate of Hopeite‐like phase (Co‐H) on NiCo‐layered double hydroxide (NiCo‐LDH) to prepare a self‐supported electrode (Co‐H/NiCo@NF). The Co‐H formed over the NiCo‐LDH shows a unique sea urchin‐like morphology. The Co‐H/NiCo@NF displays excellent HER and OER activity, requiring only overpotentials of 180 mV and 350 mV to deliver 100 mA cm−2, consequently, be capable to generate an appealing cell potential of 1.76 V for overall water splitting at the corresponding current density. In addition, the prepared Co‐H/NiCo@NF has long‐term stability against 500 mA cm−2 and exhibits a trend of increased activity, which may associate with the structural reconstructions to form new phases and the strong bonding between layers. These findings demonstrate that the multi‐layer architecture with fine‐component modulation is a promising strategy for the development of robust and efficient cobalt phosphate electrocatalysts, and the role of multi‐layer in the evolution of structural reconstructions deserves further investigation.

Due to its intricate heterogeneity, high invasiveness, and poor prognosis, triple-negative breast cancer (TNBC) stands out as the most formidable subtype of breast cancer. At present, chemotherapy remains the prevailing treatment modality for TNBC, primarily due to its lack of estrogen receptors (ERs), progesterone receptors (PRs), and human epidermal growth receptor 2 (HER2). However, clinical chemotherapy for TNBC is marked by its limited efficacy and a pronounced incidence of adverse effects. Consequently, there is a pressing need for novel drugs to treat TNBC. Given the rich repository of diverse natural compounds in traditional Chinese medicine, identifying potential anti-TNBC agents is a viable strategy. This study investigated lasiokaurin (LAS), a natural diterpenoid abundantly present in Isodon plants, revealing its significant anti-TNBC activity both in vitro and in vivo. Notably, LAS treatment induced cell cycle arrest, apoptosis, and DNA damage in TNBC cells, while concurrently inhibiting cell metastasis. In addition, LAS effectively inhibited the activation of the phosphatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway and signal transducer and activator of transcription 3 (STAT3), thus establishing its potential for multitarget therapy against TNBC. Furthermore, LAS demonstrated its ability to reduce tumor growth in a xenograft mouse model without exerting detrimental effects on the body weight or vital organs, confirming its safe applicability for TNBC treatment. Overall, this study shows that LAS is a potent candidate for treating TNBC.

The terminal reservoirs of water transfer projects directly supply water for domestic, agricultural, and industrial applications, and the water quality of these reservoirs produce crucial effects on the achievement of project targets. Typically, fish assemblages are monitored as indicators of reservoir water quality, and can also be regulated for its improvement. In the present study, we compared traditional fish landing (TFL) and environmental DNA (eDNA) metabarcoding methods for monitoring fish assemblages in three terminal reservoirs of the East Route of the South-to-North Water Transfer Project, China. Results of TFL and eDNA showed similar assemblage structures and patterns of diversity and spatial distribution with obvious differences in fish composition across three examined reservoirs. Demersal and small fish were dominant in all reservoirs. In addition, a strong association between water transfer distance and assemblages and distribution of non-native fish was found. Our findings highlight the necessity of the fish assemblage monitoring and managing for water quality and revealed the impact of water diversion distance on the structure of fish assemblages and dispersal of alien species along the water transfer project.

Urban buildings are essential components of cities and an indispensable source of urban geographic information. While there are many research efforts focused on urban buildings extraction, there are few studies on large-scale urban building mapping based on satellite images. In this research, a large-scale urban building mapping scheme based on Gaofen-2 satellite (GF-2) images is proposed based on a hierarchical approach. In this hierarchical approach, urban buildings are regarded as a mixture of dense low-rise buildings (DLBs) and sparse independent buildings (SIBs) stacked in space, which are extracted by a semantic segmentation model and an instance segmentation model, respectively. In this study, GF-2 images and OpenStreetMap data were used to extract DLB using $U^2$-Net with focal loss. GF-2 images were used to extract SIB using an improved CenterMask model with a deformable convolution network and a spatial coordinate attention module. The main urban area within the 5th ring road of Beijing was selected as the study area. With the trained model, the GF-2 image tiles of Beijing input into the models to first derive coarse maps of DLB and SIB. Postprocessing optimization was performed after combining the maps. The accuracy assessment shows that the overall accuracy of large-scale urban building mapping using the hierarchical approach proposed in this article reaches 91.5%, which is 4.8% higher than that with a traditional method. Overall, the hierarchical approach proposed in this article is effective in large-scale urban building mapping and provides new application opportunities.

High-arsenic water limits the utilization and development of water resources in Tibet, and high-arsenic groundwater is one of the major sources of arsenic input to surface water in the area. In this work, the characteristics of groundwater and the source and formation of arsenic in a typical watershed in Tibet (the lower tributaries of the Angqu River) were investigated using systematic surveys, ionic ratios, Gibbs diagrams, in combination with isotopic and heat storage calculation methods. The studies show that the chemical composition of the water in the study area is mainly determined by the rock weathering of carbonate and silicate rocks. The average recharge elevation levels of hot spring water are 4874.1 m, 4058.1 m, and 4745.0 m, respectively. Deep hot water is the main source of arsenic in the spring water, and its arsenic flux accounts for 98.44–99.77% of the measured flux in the spring water.

Inflammatory bowel disease (IBD) is an idiopathic inflammatory disease affecting the gastrointestinal tract. IBD is characterized by courses of relapse and remission, and remains incurable. Although multiple factors are related to the pathogenesis of IBD, disruption of intestinal mucosa homeostasis has been proposed to be a major contributor to IBD, and abnormal activation of immune cells is key for initiation of the inflammatory response. Macrophages are the most abundant immune cells in the intestine. Once activated, they are responsible for secretion of pro-inflammatory cytokines and chemokines to attract circulating monocytes to inflammatory sites, exacerbating the inflammatory response, and leading to tissue damage. Therefore, the suppression of activated macrophages, cytokine/chemokine production, and subsequent monocyte chemotaxis possesses great potential for the treatment of IBD. In our study, we have demonstrated the inhibitory effect of Centipeda minima total extract (CME) on the activation of NF-κB, STAT3, and MAPK signaling in LPS-stimulated RAW264.7 macrophages. In addition, we identified the significant suppressive effect of CME on CCL8 expression in activated macrophages, which potentially contributed to inhibition of monocyte chemotaxis. In the DSS-induced acute colitis mouse model, we have demonstrated the suppressive effect of CME on intestinal macrophage infiltration and its ameliorative effect in IBD. Altogether, we have provided evidence of the therapeutic effect of CME in IBD and the potential of CME for the treatment of IBD.

Breast cancer is the most commonly diagnosed cancer in females worldwide. Estimates from the World Health Organization (WHO) International Agency for Research on Cancer, suggest that globally, there were around 2.1 million new breast cancer cases and 627,000 deaths due to breast cancer in 2018. Among the subtypes of breast cancer, triple negative breast cancer (TNBC) is the most aggressive and carries the poorest prognosis, largest recurrence, and lowest survival rate. Major treatment options for TNBC patients are mainly constrained to chemotherapy, which can be accompanied by severe side effects. Therefore, development of novel and effective anti-cancer drugs for the treatment of TNBC are urgently required. Centipeda minima is a well-known traditional Chinese herbal medicine that has historically been used to treat rhinitis, sinusitis, relieve pain, and reduce swelling. Recent studies have shown that Centipeda minima exhibited efficacy against certain cancers, however, to date, no studies have been conducted on its effects in breast cancer. Here, we aimed to investigate the anti-cancer activity of the total extract of Centipeda minima (CME), and its underlying mechanism, in TNBC. In MDA-MB-231, we found that CME could significantly reduce cell viability and proliferation, induce apoptosis and inhibit cancer cell migration and invasion, in a dose and time-dependent manner. We showed that CME may potentially act via inhibition of multiple signaling pathways, including the EGFR, PI3K/AKT/mTOR, NF-κB, and STAT3 pathways. Treatment with CME also led to in vitro downregulation of MMP-9 activity and inhibition of metastasis. Further, we demonstrated that CME could significantly reduce tumor burden in MDA-MB-231 xenograft mice, without any appreciable side effects. Based on our findings, CME is a promising candidate for development as a therapeutic with high efficacy against TNBC.

The hypoglycaemic target of empagliflozin (EMP), as a novel inhibitor of sodium-glucose cotransporter (SGLT2), is clear. However, recent studies have shown that EMP also has an important role in lipid metabolism and cardiovascular diseases. The liver plays an important role in the development of type 2 diabetes (T2D), although whether EMP affects liver glucose metabolism is currently not reported. This study was designed to evaluate the effect of EMP on hepatic glucose metabolism in T2D and the underlying mechanism. A model of T2D was established by a high-fat and glucose diet (HFD) combined with streptozotocin (30 mg/kg) in male Wistar rats. Serum samples were collected to measure biochemical indicators, and liver samples were extracted for RNA-seq assay. Quantitative real-time PCR (qPCR) was used to further verify the gene expression levels detected by the RNA-seq assay. The EMP group showed significantly decreased blood glucose, triglyceride, cholesterol, non-esterified fatty acid and low-density lipoprotein cholesterol levels, and increased high-density lipoprotein cholesterol levels in serum compared with the type 2 diabetes model (MOD) group. Furthermore, EMP decreased the levels of inflammatory factors IL-1β, IL-6, and IL-8 in the serum compared to the MOD. Liver transcriptome analysis showed EMP affects a large number of upregulated and downregulated genes. Some of these genes are novel and involve in the metal ion binding pathway and the negative regulation of transcription from the RNA polymerase II promoter pathway, which are also closely related to glucolipid metabolism and insulin signaling. Our study provides new knowledge about the mechanism through which SGLT inhibitor can offer beneficial effects in T2D and especially in the hepatic metabolism. These genes found in this study also laid a solid foundation for further research on the new roles and mechanisms of EMP.

Nasopharyngeal carcinoma (NPC) is one of the most common malignant cancers in Southeast Asia and Southern China. Centipeda minima extract (CME) had previously demonstrated anti-cancer effects in human NPC. Brevilin A, a sesquiterpene lactone isolated from C. minima, has been reported to exhibit biological activities. In this study, we investigated its anti-NPC effect and further explored its molecular mechanisms. The effects of brevilin A were tested in the NPC cell lines CNE-1, CNE-2, SUNE-1, HONE1, and C666-1. Effects of brevilin A on cell viability were determined by MTT assay, and cell cycle and apoptosis were detected by flow cytometry. The molecular mechanism of cell cycle regulation and apoptosis were investigated via Western blot. Results showed that brevilin A inhibited NPC cell viability in a concentration- and time-dependent manner. Brevilin A induced cell cycle arrest at G2/M and induced apoptosis. Western blot results demonstrated that brevilin A could down-regulate cyclin D3, cdc2, p-PI3K, p-AKT, p-mTOR, and p-STAT3, while up-regulating cleaved PARP, cleaved caspase 9, and Bax. Regulation of cyclin B1, cdk6, and Bcl-2 expression by brevilin A showed dynamic changes according to dose and time. In the tumor xenograft model, brevilin A could reduce tumor growth, at a similar magnitude to cisplatin. However, notably, whereas cisplatin treatment led to significant weight loss in treated mice, treatment with brevilin A did not, indicating its relative lack of toxicity. Taken together, brevilin A regulated cell cycle, activated the caspase signaling pathway, and inhibited PI3K/AKT/mTOR and STAT3 signaling pathways in vitro, and exhibited similar efficacy to the common chemotherapeutic cisplatin in vivo, without its associated toxicity. These findings provide a framework for the preclinical development of brevilin A as a chemotherapeutic for NPC.

Building extraction from high spatial resolution remote sensing images is a hot spot in the field of remote sensing applications and computer vision. This paper presents a semantic segmentation model, which is a supervised method, named Pyramid Self-Attention Network (PISANet). Its structure is simple, because it contains only two parts: one is the backbone of the network, which is used to learn the local features (short distance context information around the pixel) of buildings from the image; the other part is the pyramid self-attention module, which is used to obtain the global features (long distance context information with other pixels in the image) and the comprehensive features (includes color, texture, geometric and high-level semantic feature) of the building. The network is an end-to-end approach. In the training stage, the input is the remote sensing image and corresponding label, and the output is probability map (the probability that each pixel is or is not building). In the prediction stage, the input is the remote sensing image, and the output is the extraction result of the building. The complexity of the network structure was reduced so that it is easy to implement. The proposed PISANet was tested on two datasets. The result shows that the overall accuracy reached 94.50 and 96.15%, the intersection-over-union reached 77.45 and 87.97%, and F1 index reached 87.27 and 93.55%, respectively. In experiments on different datasets, PISANet obtained high overall accuracy, low error rate and improved integrity of individual buildings.

Image segmentation has made great progress in recent years, but the annotation required for image segmentation is usually expensive, especially for remote sensing images. To solve this problem, we explore semi-supervised learning methods and appropriately utilize a large amount of unlabeled data to improve the performance of remote sensing image segmentation. This paper proposes a method for remote sensing image segmentation based on semi-supervised learning. We first design a Consistency Regularization (CR) training method for semi-supervised training, then employ the new learned model for Average Update of Pseudo-label (AUP), and finally combine pseudo labels and strong labels to train semantic segmentation network. We demonstrate the effectiveness of the proposed method on three remote sensing datasets, achieving better performance without more labeled data. Extensive experiments show that our semi-supervised method can learn the latent information from the unlabeled data to improve the segmentation performance.

The reproductive biology of three yellow catfish congeners was studied in the Three Gorges Reservoir of the Yangtze River, China. We compared reproductive traits among the lentic, transitional, and lotic zones. A total of 4502 individuals of the three species was collected, and the sex ratio, size at 50% maturity, spawning season, fecundity, and egg size were determined. Results showed that populations inhabiting the lotic zone spawned earlier than those inhabiting the lentic zone. For the three species, fecundities were significantly higher for populations in the lotic zone than for those in the lentic and transitional zones (P < 0.05). Pelteobagrus vachelli (Richardson) and P. fulvidraco (Richardson) displayed an obvious trade-off between egg size and fecundity, whereas P. nitidus (Sauvage et Dabry) produced the largest eggs in the lotic zone. Sex ratios were significantly different among zones (P < 0.05, for each species), but the bias patterns were different. Sizes at 50% maturity of female P. nitidus and P. vachelli were the largest in the lotic zone and the smallest in the transitional zone, but was similar among zones for P. fulvidraco. Overall results suggest that the three yellow catfish species developed different reproductive traits among the three habitats in the TGR, whereas the variations reflected further interspecific differences. Our study indicates the importance of riverine habitats for the conservation of species of fish, even for species such as these eurytopic catfish inhabiting the upper reach of the Yangtze River. This study further suggests that species-specific responses should be considered when evaluating the influences of new hydropower projects, even for such closely related species of fish.

Nasopharyngeal carcinoma (NPC) is a high morbidity and mortality cancer with an obvious racial and geographic bias, particularly endemic to Southeast China. Our previous studies demonstrated that Centipeda minima extract (CME) exhibited anti-cancer effects in human NPC cell lines. Arnicolide C and arnicolide D are sesquiterpene lactones isolated from Centipeda minima. In this study, for the first time, we investigated their anti-NPC effects and further explored the related molecular mechanisms. The effects of both arnicolide C and arnicolide D were tested in NPC cells CNE-1, CNE-2, SUNE-1, HONE1, and C666-1. The results showed that the two compounds inhibited NPC cell viability in a concentration- and time-dependent manner. As the inhibitory effect of arnicolide D was the more pronounced of the two, our following studies focused on this compound. Arnicolide D could induce cell cycle arrest at G2/M, and induce cell apoptosis. The molecular mechanism of cell cycle regulation and apoptosis induction was investigated, and the results showed that arnicolide D could downregulate cyclin D3, cdc2, p-PI3K, p-AKT, p-mTOR, and p-STAT3, and upregulate cleaved PARP, cleaved caspase 9, and Bax. Regulation of cyclin B1, cdk6, and Bcl-2 expression by arnicolide D showed dynamic changes according to dose and time. Taken together, arnicolide D modulated the cell cycle, activated the caspase signaling pathway, and inhibited the PI3K/AKT/mTOR and STAT3 signaling pathways. These findings provide a solid base of evidence for arnicolide D as a lead compound for further development, and act as proof for the viability of drug development from traditional Chinese medicines.

Pyrrosiae Folium (PF) is a commonly used Chinese herb medicine originating from three Pyrrosia species for the treatment of urinary infection and urolithiasis. According to Chinese medicine practice, different specie origins led to some variations in the therapeutic effects of PF. To ensure the safety and efficacy of PF in clinical practice, it is necessary to establish a reliable and integrative method to distinguish PF occurring from the three species. In the present paper, a HPLC–DAD method was developed and applied to simultaneously analyze five major compounds in PF. Afterwards, multivariate statistical analyses including principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were applied for specie discrimination and integrative quality evaluation based on quantitative data. The chemical determination and pattern recognition results of 35 batches of PF samples indicated that PF samples from three species showed different chemical profiles and could be discriminated clearly. In conclusion, the present method is rapid and reliable for the quality assessment and species discrimination of PF.

Bufalin (BFL) has excellent physiological activities such as defending tumors, improving cardiac function, and so on. However, due to its poor water-solubility and bioavailability, the clinical application of BFL remains limited. In order to improve bioavailability of BFL, in our previous research, a novel peptide-dendrimer (PD) was synthesized and applied to encapsulate BFL. In the present study, we investigate the absorption property and mechanism of BFL in free form and BFL-peptide-dendrimer inclusion (BPDI) delivery system by using the Caco-2 cell monolayer model in vitro. The apparent permeability coefficient (Papp) values of BFL in free or BPDI form were over 1.0 × 10−6 cm/s. Meanwhile, their almost equal bi-directional transport and linear transport percentage with time and concentration course indicated that BFL in both forms was absorbed mainly through passive diffusion. The most important result is that the Papp values of BFL increased about three-fold more BPDI than those of its free form, which indicated the intestinal permeability of BFL could be improved while BFL was encapsulated in BPDI form. Therefore, PD encapsulation may be a potential delivery system to increase the bioavailability of BFL.

A simple, accurate and reliable high performance liquid chromatography coupled with photodiode array detection (HPLC-DAD) method was developed and then successfully applied for simultaneous quantitative analysis of eight compounds, including chlorogenic acid (1), (R/S)-flavanomarein (2), butin-7-O-β-d-glucopyranoside (3), isookanin (4), taxifolin (5), 5,7,3′,5′-tetrahydroxyflavanone-7-O-β-d-glucopyranoside (6), marein (7) and okanin (8), in 23 batches of snow chrysanthemum of different seed provenance and from various habitats. The results showed total contents of the eight compounds in the samples with seed provenance from Keliyang (Xinjiang, China), are higher than in samples from the other five provenances by 52.47%, 15.53%, 19.78%, 21.17% and 5.06%, respectively, which demonstrated that provenance has a great influence on the constituents in snow chrysanthemum. Meanwhile, an ultra performance liquid chromatography coupled with electrospray ionization and quadrupole time-of-flight-mass spectrometry (UPLC-ESI-QTOF-MS) was also employed to rapidly separate and identify flavonoids and phenolic acids in snow chrysanthemum from Keliyang. As a result, a total of 30 constituents, including 26 flavonoids and four phenolic acids, were identified or tentatively identified based on the exact mass information, the fragmentation characteristics, and retention times of eight reference standards. This work may provide an efficient approach to comprehensively evaluate the quality of snow chrysanthemum.