Ang-1 promotes tumorigenesis and mediates the anti-cancer effects of Artesunate on Choroidal melanoma via the regulation of Akt/mTOR signaling pathway.

The impact of Ang-1 on tumors remains a subject of contention, with its mechanism of action exhibiting complexity in the progression of diverse tumor types. Ang-1 has been shown to promote the progression of glioma, glioma, esophageal and human cervical cancer, whereas it exerts inhibitory effects on the growth of breast and colon cancer. However, the specific function of Ang-1 in CM has not been clarified. This research aims to explore the function of Ang-1 on CM and the underlying mechanism. WB and qPCR were utilized to measure the expression levels of different factors in CM cells. Clonogenic, CCK-8 and Transwell migration assay were used to probe CM cells' proliferation and migration ability. Xenograft tumor model was used to testify the effect of Ang-1 and Artesunate (ART) on the growth of CM in vivo. We found Ang-1 promoted CM proliferation and migration, while it was inhibited by ART in vitro. Moreover, both ART treatment and Ang-1 knockdown had the effect of suppressing tumor growth in CM xenograft model. Mechanically, Ang-1 activated Akt/mTOR pathway and induced epithelial-mesenchymal transition (EMT) in CM cells. Furthermore, ART regulated Akt/mTOR pathway by decreasing the expression of Ang-1 in CM cells. Ang-1 promotes tumorigenesis of CM by regulating Akt/mTOR pathway, which can be inhibited by ART.

Integrating network pharmacology, molecular docking and experimental verification to reveal the mechanism of artesunate in inhibiting choroidal melanoma.

Background: Artesunate (ART), a natural compound derived from Artemisia annua, has shown promising clinical potentials in the treatment of various tumors, but the exact mechanism is unclear. Choroidal melanoma (CM) is a major malignant ocular tumor in adults, known for its significant malignancy and poor prognosis, with limited efficacy in current treatments. This study explored the anti-CM effects and mechanisms of ART using a combination of network pharmacology, molecular docking and experimental validation. Methods: Potential targets of ART were screened in PubChem, Swiss Target Prediction and Traditional Chinese Medicine Systems Pharmacology (TCMSP) Database Analysis Platform databases, while target genes related to CM prognosis were selected from Online Mendelian Inheritance in Man (OMIM), GeneCards and DisGeNET databases. The intersection of these two groups of datasets yielded the target genes of ART involved in CM. Protein-protein interaction (PPI) network analysis of the intersecting targets, as well as Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, were conducted to identify core targets and critical pathways. Molecular docking methods were performed to predict the binding interactions between ART and core targets. The effects of ART on CM were evaluated through CCK8, colony formation, transwell, as well as flow cytometry assays to detect apoptosis, cell cycle, reactive oxygen species (ROS). Western blot (WB) assays were conducted to investigate the impact of ART on key proteins and pathways associated with CM. Finally, in vivo assays were conducted to further validate the effects of ART on subcutaneous tumors in nude mice. Results: Research has shown that key pathways and core targets for ART in treating CM were identified through a network pharmacology approach. Molecular docking results verified the strong binding affinity between ART and these core targets. The analysis and predicted results indicated that ART primarily exerted its effects on CM through various tumor-related pathways like apoptosis. The assays in vitro confirmed that ART significantly inhibited the proliferation and migration of CM cells. This was achieved by promoting apoptosis through activation of the p53 signaling pathway, causing cell cycle arrest at the G0/G1 phase by inhibiting the PI3K/AKT/mTOR signaling pathway and increasing the intracellular level of ROS by activating the NRF2/HO-1 signaling pathway. Additionally, the assays in vivo further validated the significant proliferation-inhibitory effect of ART on CM. Conclusion: This study, making the initial exploration, illustrated through network pharmacology combined with molecular docking and in vitro/in vivo assays, confirmed that ART exerted potential anti-cancer effects on CM by promoting apoptosis, inducing cell cycle arrest and increasing intracellular levels of ROS. These findings suggested that ART held significant therapeutic potential for CM.

Artesunate inhibits vasculogenic mimicry in choroidal melanoma through HIF-1 α/ VEGF/PDGF pathway.

Choroidal melanoma (CM), a highly metastatic eye tumor, exhibits vasculogenic mimicry (VM) facilitated by hypoxia-induced angiogenesis. This study explored the inhibitory impact of the anti-malarial drug Artesunate (ART) on CM VM through modulation of the HIF-1α/VEGF/PDGF pathway. Immunohistochemistry (IHC) confirmed VM in CM with elevated VEGF and PDGF expression. Hypoxia promoted CM proliferation, upregulating HIF-1α, VEGF and PDGF. VEGF and PDGF enhanced CM migration, invasion and VM, with HIF-1α playing a crucial role. ART mitigated VM formation by suppressing the HIF-1α/VEGF/PDGF pathway, highlighting its potential as an anti-tumor agent in CM.

Artesunate attenuates the tumorigenesis of choroidal melanoma via inhibiting EFNA3 through Stat3/Akt signaling pathway.

PURPOSE: Choroidal melanoma (CM), a kind of malignant tumor, is the main type of Uveal melanoma and one half of CM patients develop metastases. As a member of Eph/ephrin pathway that plays vital role in tumors, EphrinA3 (EFNA3) has been proved to promote tumorigenesis in many tumors. But the effect of EFNA3 in CM has not been studied yet. Through inhibiting angiogenesis, inducing apoptosis and autophagy and so on, Artesunate (ART) plays a key anti-tumor role in many tumors, including CM. However, the exact mechanisms of anti-tumor in CM remain unclear. METHODS: The UALCAN and TIMER v2.0 database analyzed the role of EFNA3 in CM patients. Quantitative real time polymerase chain reaction (qPCR) and Western blot were used to detect the expression of EFNA3 in CM. The growth ability of CM was tested by clonogenic assay and Cell counting kit-8 assay, and the migration ability using Transwell assay. RESULTS: Our results found EFNA3 boosted CM cells' growth and migration through activating Stat3/Akt signaling pathway, while ART inhibited the tumor promoting effect of CM via downregulating EFNA3. In xenograft tumor model, EFNA3 knockdown and ART significantly inhibited tumor growth. CONCLUSION: EFNA3 could be a valuable prognostic factor in CM.

Yinhuang granule alleviates carbon tetrachloride-induced liver fibrosis in mice and its mechanism.

BACKGROUND: Liver fibrosis is a formidable global medical challenge, with no effective clinical treatment currently available. Yinhuang granule (YHG) is a proprietary Chinese medicine comprising Scutellariae Radix and Lonicerae Japonicae Flos. It is frequently used for upper respiratory tract infections, pharyngitis, as well as acute and chronic tonsillitis. AIM: To investigate the potential of YHG in alleviating carbon tetrachloride (CCl4)-induced liver fibrosis in mice. METHODS: To induce a hepatic fibrosis model in mice, this study involved intraperitoneal injections of 2 mL/kg of CCl4 twice a week for 4 wk. Meanwhile, liver fibrosis mice in the low dose of YHG (0.4 g/kg) and high dose of YHG (0.8 g/kg) groups were orally administered YHG once a day for 4 wk. Serum alanine/aspartate aminotransferase (ALT/AST) activity and liver hydroxyproline content were detected. Sirius red and Masson's trichrome staining assay were conducted. Real-time polymerase chain reaction, western-blot and enzyme-linked immunosorbent assay were conducted. Liver glutathione content, superoxide dismutase activity level, reactive oxygen species and protein carbonylation amount were detected. RESULTS: The administration of YHG ameliorated hepatocellular injury in CCl4-treated mice, as reflected by decreased serum ALT/AST activity and improved liver histological evaluation. YHG also attenuated liver fibrosis, evident through reduced liver hydroxyproline content, improvements in Sirius red and Masson's trichrome staining, and lowered serum hyaluronic acid levels. Furthermore, YHG hindered the activation of hepatic stellate cells (HSCs) and ameliorated oxidative stress injury and inflammation in liver from CCl4-treated mice. YHG prompted the nuclear accumulation of nuclear factor erythroid 2-related factor 2 (Nrf2) and upregulated the expression of Nrf2-dependent downstream antioxidant genes. In addition, YHG promoted mitochondrial biogenesis in liver from CCl4-treated mice, as demonstrated by increased liver adenosine triphosphate content, mitochondrial DNA levels, and the expression of peroxisome proliferator-activated receptor gamma coactivator 1 alpha and nuclear respiratory factor 1. CONCLUSION: YHG effectively attenuates CCl4-induced liver fibrosis in mice by inhibiting the activation of HSCs, reducing inflammation, alleviating liver oxidative stress damage through Nrf2 activation, and promoting liver mitochondrial biogenesis.

Intravitreal injection of conbercept for diabetic macular edema complicated with diabetic nephropathy.

AIM: To observe the therapeutic effect of conbercept on diabetic macular edema (DME) complicated with diabetic nephropathy (DN). METHODS: In this retrospective study, 54 patients (54 eyes) that diagnosed as DME from January 2017 to October 2021 were collected. The patients were divided into two groups: DME patients with DN (25 eyes), and DME patients without DN (29 eyes). General conditions were collected before treatment, laboratory tests include fasting blood glucose, HbA1c, microalbumin/creatinine, serum creatinine. Optical coherence tomography (OCT) was used to check the ellipsoidal zone (EZ) and external limiting membrane (ELM) integrity. Central macular thickness (CMT), best corrected visual acuity (BCVA), and retinal hyperreflective foci (HF) as well as numbers of injections were recorded. RESULTS: There were significant differences between fasting blood glucose, HbA1c, serum creatinine, urinary microalbumin/creatinine, and estimated glomerular filtration rate (eGFR) between the two groups (all P<0.05). EZ and ELM continuity in the DME+DN group was worse than that in the DME group (P<0.05). BCVA (logMAR) in the DME group was significantly better than that in the DME+DN group at the same time points during treatment (all P<0.05). CMT and HF values were significantly higher in the DME+DN group than that in the DME group at the all time points (all P<0.05) and significantly decreased in both groups with time during treatment. At 6mo after treatment, the mean number of injections in the DME+DN and DME group was 4.84±0.94 and 3.79±0.86, respectively. CONCLUSION: Conbercept has a significant effect in short-term treatment of DME patients with or without DN, and can significantly ameliorate BCVA, CMT and the number of HF, treatment efficacy of DME patients without DN is better than that of DME patients with DN.

Modeling Spatio-Temporal Variations in the Habitat Utilization of Swordtip Squid (Uroteuthis edulis) in the East China Sea and Southern Yellow Sea.

Accurately modeling the distribution of keystone species is of utmost importance to gain a comprehensive understanding of their complex ecological dynamics and to develop effective strategies for sustainable scientific management. In the coastal China ecosystem, the swordtip squid (Uroteuthis edulis) stands out as a keystone species with significant commercial and ecological value. Despite its importance, research on the ecological dynamics of this species remains limited and requires further investigation. To investigate the spatial and temporal variability in the distribution of U. edulis and identify the key environmental drivers in the East China Sea (ECS) and southern Yellow Sea across different seasons, we generated ensemble models using oceanographic variables and fishery-independent scientific survey data collected from 2016 to 2018. Our results revealed that U. edulis predominantly inhabited the central and southern regions of the ECS throughout the year. The primary environmental variables driving its distribution varied by season, with the sea surface temperature being the most important in spring, sea surface height in summer and autumn, and depth in winter. During summer and autumn, the suitable habitats of U. edulis were found to be largest and extended northwards towards the coastline. However, they migrated southwards to the waters near the edge of the ECS continental shelf with smaller suitable areas in the spring and winter. These results suggested that U. edulis exhibited season-specific habitat preferences and responded to changing environmental conditions throughout the year. The observed seasonal distribution patterns were likely influenced by the fluctuating mixture of waters (ocean currents) from different sources, with varying physical and chemical characteristics throughout the year. Our study provides baseline data for comprehending the population dynamics of U. edulis and highlights the significance of considering species' habitat preferences in a dynamic environment.

[Spatiotemporal distribution of Lophius litulon in the southern Yellow Sea and East China Sea]. / 黄海南部及东海黄鮟鱇时空分布.

In order to investigate the temporal-spatial distribution of yellow goosefish in the open waters of the southern Yellow Sea (SYS) and East China Sea (ECS), according to the fishery data by bottom-trawl surveys and environmental data including sea bottom temperature (SBT), sea bottom salinity (SBS), bottom concentration of dissolved oxygen (BDO) and depth during 2018-2019, we established habitat suitability index (HSI) models by arithmetic mean (AMM) and geometric mean (GMM) methods, and compared the model outputs by cross validations. Especially, the weight of each environmental factor was evaluated by boosted regression tree (BRT). Results showed that the area with the highest habitat quality varied among seasons. The yellow goosefish mainly inhabited in the adjacent area of the Yangtze River Estuary and coastal waters of Jiangsu Province with depth ranged from 22 to 49 m in spring. The optimal inhabitation was located in the SYS, with the bottom temperature ranged from 8.9-10.9 ℃ in summer and autumn. Particularly, the optimal inhabitation extended from the SYS to the ECS with bottom temperature ranged from 9.2-12.7 ℃ in winter. Results of BRT models showed that depth was the most important environmental factor in spring and bottom temperature was the crucial one in the other three seasons. Results of cross validation showed that the weighted AMM-based HSI model performed better for yellow goosefish in spring, autumn and winter. The distribution of yellow goosefish was closely related to its biological traits and environmental factors in the SYS and ECS, China.

Trophic niche and gut microbiota of Dosidicus gigas in the eastern equatorial Pacific Ocean.

Understanding the adaptation of important marine species to environmental changes is critical for evaluating the effects of fisheries and climate change on marine services. The jumbo squid, Dosidicus gigas, is a keystone species in the eastern Pacific, which plays an intermediate role in the marine food web. Better understanding of their responses to climate change would be a big step to understand their population dynamics. In this study, stable isotope and high-throughput 16S rRNA sequencing were used to compare the variation of trophic niche, gut length, and gut microbiota of D. gigas in the eastern equatorial water during normal and El Niño periods. The results showed a significant variation in δ13C values for D. gigas in different periods, indicating differences in their food sources. The main phylum-level gut microbiome included Actinobacteria, Bacteroidetes, Cyanobacteria, Firmicutes, Proteobacteria, and Tenericutes. There was no significant difference in the gut microbial composition during normal and El Niño periods, but differences in gut microbial diversity and relative abundance of some phyla bacteria. El Niño events could decrease the trophic niche breadth of D. gigas, and positively impact gut length and gut microbial diversity and richness. Firmicutes, Bacteroidetes, Spirochaetes, WPS-2, and Kiritimatiellaeota had a significant increase in the gut microbiota of D. gigas combined with a more concentrated intraspecific rank of distance during El Niño, suggesting that the changes of habitat and food sources caused by El Niño events could limit the distribution range of D. gigas. D. gigas might change their digestive system to improve the digestive and absorption capacity and enhance their immunocompetence. Such a climate-driven alteration might help D. gigas rapidly adapt to the changes of environmental conditions and food resources under El Niño.

An evaluation of observer monitoring program designs for Chinese tuna longline fisheries in the Pacific Ocean using computer simulations.

This paper evaluates the performance of different observer coverage rates and 9 possible sampling designs to estimate via computer simulation the total catch of target and non-target species for Chinese tuna longline fisheries in the Pacific Ocean. The stratified random samplings include different stratification schemes (based on target species or fishing areas) with different strategies for allocating observers. The observer data from 103 vessels between 2010 and 2019 were assumed to be the "true" sampling population. We concluded that the accuracy of catch estimates had a significant positive relationship with species detectability and observer coverage rate. On average, the accuracy improved by 50% when the coverage rate increases from 5 to 20%. Current simple random sampling in Chinese tuna longline fisheries is less efficient for monitoring many species. Stratified sampling designs based on the target species tended to yield the most accurate estimates of the total catch. Allocating the observers based on the scale of the fleets in different stratum seemed to be less efficient. The proportion of observers between different fleets should be adjusted according to different monitoring objectives. In general, a large proportion of observers are recommended to be allocated onboard vessels targeting bigeye tuna (Thunnus obesus). This study has the potential to have a significant contribution to future designs of the observer monitoring programs in Chinese tuna longline fishery and many other fisheries.

Design and application of an open tubular capillary reactor for solid-phase permethylation of glycans in glycoprotein.

The permethylation derivatization method for structural analysis of glycans is important for characterizing glycoproteins in the study of glycomics. An open tubular capillary reactor coated with NaOH was designed and constructed for solid-phase permethylation of glycans in glycoproteins. The flow rate, capillary length, and inner diameter of the reactor were optimized. The permethylation rate of the model sample ß-cyclodextrin reached 81% with a flow rate of 1 µL min(-1) in a 32 cm long capillary reactor (i.d. 500 µm). A trace amount of mucin O-glycans was permethylated by the open tubular reactor under low pressure without interference from the freezing of DMSO. Analysis indicates that using this open tubular reactor is a fast, convenient, and efficient method for the permethylation of protein O-glycan.

[Protein enrichment based on large volume recycling injection].

Biological proteins have great differences in size, hydrophobicity, pH and relative abundance. Especially, the relative contents of high-abundance proteins and low abundance proteins can reach 11 orders of magnitude or more in blood. However, the contents of proteins which have an important impact on the biological function and have related to major diseases are usually very low in body fluids or tissues. So, it is very meaningful to detect the low abundance proteins in biological samples. A method of large volume recycling injection was developed as a new sample enrichment technology in this study. The low absolute contents of pro- teins in a sample were improved by increasing the volume of sample, and the enrichment efficiency was also increased with the addition of injection times. With the pork liver protein as an example, under the optimized conditions, the representative contents which were no peaks or little peaks and more peaks with low signals were studied. The contents of peaks at the retention times of 11.38 min and 12.58 min were enriched by 11 time injections of 500 µL each time. The enrichment factors respectively reached 52 and 61 which were closed to theoretical values. This method can be used for the enrichment of biological protein samples.