Xanthones from Gentianella acuta (Michx.) Hulten Ameliorate Colorectal Carcinoma via the PI3K/Akt/mTOR Signaling Pathway.

Colorectal carcinoma (CRC) is a kind of malignant tumor closely related to ulcerative colitis. Xanthone derivatives are one of the most promising therapeutic drugs which have been used in phase I/II clinical trials for cancer therapy. Our previous study indicated that the aerial parts of Gentianella acuta Michx. Hulten (GA) was rich in xanthones and showed a good therapeutic effect on ulcerative colitis in mice, suggesting that GA xanthones might have some therapeutic or ameliorative effects on CRC. However, no relevant study has been reported. This study aims to find the effective substances of GA inhibiting CRC and clarify their mechanism. Solvent extraction, column chromatographic separation, and LC-MS analysis were used to characterize the 70% EtOH extract of GA and track xanthones abundant fraction XF. MTT assay was carried out to clarify the activity of GA fractions; the result showed XF to be the main active fraction. LC-MS analysis was executed to characterize XF, 38 xanthones were identified. Network pharmacology prediction, in vitro activity screening, and molecular docking assay were combined to predict the potential mechanism; the PI3K/Akt/mTOR signaling pathway was found to be most important. Western blot assay on the main active xanthones 1,3,5-trihydroxyxanthone (16), 1,3,5,8-tetrahydroxyxanthone (17), 1,5,8-trihydroxy-3-methoxyxanthone (18), and 1,7-dihydroxy-3,8-dimethoxyxanthone (19) was used to verify the above prediction; these xanthones were found to inhibit the PI3K/Akt/mTOR signaling pathway, and 17 played a significant role among them through Western blot assay using PI3K/AKT/mTOR agonist IGF-1. In conclusion, this study demonstrated that GA xanthones were effective compounds of GA inhibiting CRC by regulating PI3K/Akt/mTOR signaling pathway transduction, at least. Importantly, 1,3,5,8-tetrahydroxyxanthone (17), the most abundant active xanthone in GA, might be a candidate drug for CRC.

[Spatial and Temporal Distribution of Antibiotic Resistance Genes in Different River Sediments in Typical Cities: A Case Study of Shijiazhuang].

Rivers are important reservoirs of antibiotic resistance genes (ARGs). However, most current studies have focused on the temporal and spatial distribution, and data on the differences in the species and abundance of ARGs between urban and rural rivers is still lacking for certain areas. In view of this, two rural rivers and three urban rivers were selected in Shijiazhuang City. In both December 2020 and April 2021, sediments were collected at 15 sampling sites. Metagenomic sequencing technology was used to compare the differences in temporal-spatial variation for ARGs in sediments. The results showed that:① 162 and 79 ARGs were detected in urban (4 776 ±4 452) and rural rivers (1 043 ±632), respectively. The abundance and species of ARGs in urban rivers were higher than those in rural rivers. ② The relative abundances of sulfonamide (SAs,27 %), aminoglycoside (AGs,26 %), and multidrug (MDs,15 %) ARGs had the highest abundance in urban rivers, whereas the relative abundance of MDs ARGs was highest in rural rivers (65 %). On the whole, the complexity of ARGs in urban rivers was higher than that in rural rivers. ③ There was a significant positive correlation between SAs, AGs, MDs, tetracycline, phenicol, macrolides-lincosamids-streptogramins (MLS), ß-lactams, and diaminopyrimidine ARGs in urban rivers (P < 0.01); however, there was a significant negative correlation between glycopeptide ARGs and all types of ARGs (P < 0.05 and P < 0.01). There was a significant positive correlation between MDs and SAs ARGs in rural rivers (P < 0.05), but there was a significant negative correlation between amino aminocoumarin, peptide, rifamycin, and fosfomycin ARGs (P < 0.05 and P < 0.01). ④ For the temporal variation in urban rivers, 162 ARGs (4 776 ±4 452) and 148 ARGs (5 673 ±5 626) were detected in December and April, respectively. For the temporal variation in rural rivers, 79 species (1 043 ±632) and 46 species (467 ±183) were detected in December and April, respectively. ⑤ RDA analysis results showed that the spatial-temporal distributions of ARGs in urban and rural rivers were different. Correlation analysis showed that the ARGs in urban rivers were significantly correlated with the number of industrial enterprises, whereas the ARGs in rural rivers were significantly correlated with the output value of animal husbandry. In general, this study identified the main influencing factors for ARGs in different rivers and provided data support for ARGs risk management in different rivers.

[Antibiotic Pollution and Its Effects on the Spatiotemporal Variation in Microbial Community Structure and Functional Genes in Sediment of Baiyangdian Lake].

Microbial communities play an important role in water quality regulation and biogeochemical cycles in lakes, and their community structure and function are affected by environmental factors. Therein, antibiotics affect the abundance, diversity, composition, and function of microbial communities. In this study, Baiyangdian Lake was selected as the study area. Sediment samples of 16 sites were collected in August 2018 and April 2019, respectively. Ultra-high performance liquid chromatography-mass spectrometry （HPLC-MS/MS） was used to determine the content of typical antibiotics-quinolones （QNs） in sediment. Through high-throughput sequencing technology, the structure and function of microbial communities was analyzed in the sediments to explore the spatiotemporal variation. Thereinto, redundancy analysis （RDA） was used to identify the key influencing factors of spatiotemporal variation of microbial communities. The results showed that： ① From August to April, the average ω（QNs） showed an increasing trend, and its mean value changed from 3.91 ng·g-1 to 6.34 ng·g-1, with significant seasonal differences in oxolinic acid （OXO） and total QNs content （P < 0.05）. ② In terms of temporal variation, the dominant bacteria were Proteobacteria and Chloroflexi. The relative abundance of Proteobacteria showed a decreasing trend, whereas Chloroflexi showed an increasing trend； at the genus level, the dominant bacteria genera in August were norank_ f__Anaerolineaceae and Thiobacilus, and the dominant bacteria genera in April were Acinetobacter and norank_ f_Anaerolineaceae, and the dominant bacteria genera had significant seasonal differences （P < 0.05）. ③ In terms of temporal variation, the index of Simpson, Chao, Ace, and OTU number all showed an increasing trend, and the seasonal differences were significant （P < 0.05）. ④ In terms of spatial variation, there were no significant spatial differences among functional genes of COG. In terms of temporal variation, there were significant seasonal differences in functional genes of energy production and conversion, carbohydrate transport and metabolism, transcription, cell wall/membrane/envelope biogenesis, and signal transduction mechanisms （P < 0.01 and P < 0.05）. ⑤ Microbial community structure and functional genes were significantly correlated with QNs （P < 0.01 and P < 0.05）, and QNs were the main influencing factors. Therefore, QNs were the main factor affecting the changes in microbial community structure and functional genes in sediments of Baiyangdian Lake. Thus, comprehensive control of antibiotic pollution in sediments should be further strengthened.

[Spatial-temporal Distribution and Risk Assessment of Quinolones Antibiotics in Soil of Shijiazhuang City].

Due to their importance in human medicine, quinolones (QNs), as a typical class of antibiotics, are considered to be the "highest priority critically important antimicrobials" by the World Health Organization (WHO). In order to clarify the spatial-temporal variation and risk of QNs in soil, 18 representative topsoil samples were respectively collected in September 2020 (autumn) and June 2021 (summer). The contents of QNs antibiotics in soil samples were determined using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), and the ecological risk and resistance risk were calculated using the risk quotient method (RQ). The results showed that:① the average content of QNs decreased from autumn to summer (the average contents of QNs were 94.88 µg·kg-1in autumn and 44.46 µg·kg-1 in summer); the highest values appeared in the middle area. ② The average proportion of silt was without change, whereas the average proportion of clay and sand was increased and decreased, respectively; the average contents of total phosphorus (TP), ammonia nitrogen (NH4+-N), and nitrate nitrogen (NO3--N) also decreased. ③ The content of QNs was significantly correlated with soil particle size, nitrite nitrogen (NO2--N), and nitrate nitrogen (NO3--N) (P<0.05). ④ The combined ecological risk of QNs showed high risk level (RQsum>1), whereas the combined resistance risk of QNs showed medium risk level (0.1

[Spatial-temporal Changes and Driving Factors of Soil Microbial Communities in a Typical City of North China Plain].

The structure and function of microbial communities are affected by several environmental factors. To clarify the spatial-temporal changes and main influencing factors of soil microbial communities in a typical pharmaceutical city, it is urgent to study the spatial-temporal changes in microbial communities in soils for typical cities. Shijiazhuang City was selected as the study area, and 12 sampling sites were selected. The topsoil was collected in June (summer) and September (autumn) of 2021. The 16S rRNA high-throughput sequencing technology was used to study the structure and function of microbial communities in the soil and explore their spatial-temporal changes. Concurrently, Pearson correlation analysis was applied to establish the correlation between the microbial community and environmental factors, and identify the main driving factors of temporal and spatial changes in the microbial community. The results showed that:① Actinobaciota and Proteobateria were the main dominant bacteria in the surface soil of Shijiazhuang City; at the phylum level, the relative abundance of Actinobacteria and Proteobateria decreased from summer to autumn; at the genus level, the dominant genera were Arthrobacter and unknown genera in summer and Arthrobacter and Candidatus_Nitrocosmicus in autumn, which showed significant seasonal differences (P<0.05). ② For seasonal variation, the mean values of the Simpson, Ace, and Chao indices increased, whereas the mean values of OTU decreased; for spatial variation, the Shannon and Simpson indices showed significant spatial difference (P<0.01 and P<0.05). ③ There were no significant spatial-temporal differences in various functional genes; thereinto, the relative abundances of energy production and transformation functional genes were the highest (24.06%-24.84% in summer and 24.63%-25.98% in autumn, respectively). ④ The compositions of microbial community, diversity index, and functional genes were significantly correlated with quinolone antibiotics (QNs), total phosphorus (TP), and nitrate nitrogen (NO3--N), most significantly correlated with QNs (|r|>0.900), which indicated that antibiotics were the main driving factor of soil microbial communities. Therefore, to ensure the stability of microbial community structure and function in urban soil, the comprehensive management and control of antibiotic pollution in soil should be further strengthened.

[Source Analysis and Health Risk Assessment of Heavy Metals in the Groundwater of Shijiazhuang, a Typical City in North China Plain].

Increasing attention has been paid to the heavy metal pollution in groundwater. The source analysis and risk assessment of heavy metals will provide data and method support for the targeted control of heavy metal pollution in groundwater. In this study, 20 sampling sites were selected in Shijiazhuang City. The APCS-MLR model and health risk model were applied to analyze and evaluate the pollution sources and health risks of 10 types of heavy metals in the groundwater of Shijiazhuang. The results showed that ① the mean concentration of heavy metals in groundwater followed the order of Fe>Zn>Mn>Cu>Al>Pb>Cr>As>Cd>Hg, and the mean ρ(Fe) and ρ(Pb) were 260.3 µg·L-1 and 10.01 µg·L-1, respectively. According to the results of the single factor and Nemerow index, Pb, Fe, and Cd primarily contributed to the heavy metal pollution in the groundwater. ② The concentration of heavy metals ranged from 47.30 to 2560 µg·L-1. In terms of spatial distribution, the highest concentration appeared at S3 (2560 µg·L-1), whereas the lowest concentration was at S9 (47.30 µg·L-1). ③ Source analysis results showed that industrial and agricultural activities, transportation emission, and geological background were the major heavy metal sources, among which the contribution of industrial and agricultural activities was the highest (47.83%). ④ The industrial-agricultural activities posed a potential threat to adults (HI>1); however, the non-cancer and the cancer risks of other sources for both adults and children were at an acceptable level (HI<1) and potential threat level, respectively; industrial-agricultural activities were the major source of non-cancer (adults:52.46%, children:52.45%) and cancer risks (adults:65.22%, children:65.69%), among which Cd and As showed high cancer risk. Therefore, to ensure the safety of the groundwater environment, strictly controlling the pollution sources and further strengthening the risk control of heavy metal pollution in groundwater are necessary.

[Source Apportionment and Source-specific Risk of Typical Antibiotics in Baiyangdian Lake].

The current situation of antibiotic pollution in lakes is critical. At present, most of the previous studies on antibiotics in lakes have focused on the spatiotemporal distribution and risk assessment, while less attention has been paid to the source apportionment. Ultra-high performance liquid chromatography-mass spectrometry was used to determine the concentration of tetracyclines (TCs), sulfonamides (SAs), and quinolones (QNs) in the samples. The source apportionment and source-specific risk of typical antibiotics in the study area were analyzed using the combination of a PMF model and risk quotients (RQ). The results showed that ① the total concentrations of target antibiotics (Σ antibiotics) ranged from ND to 2635 ng·L-1 for surface water and from ND to 259.8 ng·g-1 for sediments. ② The spatial distribution of QNs in surface water decreased from west to east, SAs decreased from middle to north and south, and TCs increased from middle to north and south. In the sediment, QNs decreased from middle to east and west, whereas SAs and TCs increased from east to west. ③ Aquaculture was the major antibiotic source, accounting for the highest proportion (33.2%), followed by sewage treatment plants (29.2%), livestock activities (18.9%), and domestic sewage (18.7%). ④ The ecological risk assessment results showed that enrofloxacin and flumequine were at a medium-high risk level. ⑤ For the spatial distribution of source-specific risk, the results showed that the aquaculture at S1 was at a high risk level, whereas the source-specific risks for other sites were at a medium-low risk level. In terms of source types, aquaculture was at a medium-high risk level, whereas the other sources were at a medium-low risk level. Therefore, considering the major sources and source-specific risk level of antibiotics, more precise and scientific antibiotic risk control should be adopted in Baiyangdian Lake.

[Distribution Coefficient of QNs in Urban Typical Water and Its Main Environmental Influencing Factors].

Sediment is the main storage medium of antibiotics in a water environment, and a growing body of research has focused on the distribution behavior of antibiotics in water-sediment. However, most of the previous studies were based on laboratory simulation, and less attention was paid to the distribution behavior of antibiotics in a natural water environment and its correlation with environmental factors. Thus, the surface water and sediment in Shijiazhuang were taken as the research object for this study. The temporal and spatial distribution characteristics of quinolone (QNs) antibiotics in Shijiazhuang water were analyzed by using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS), calculating the distribution coefficients of quinolone (QNs) antibiotics in water and sediment, and confirming the main environmental factors influencing the distribution coefficient in natural water. The results showed that:① the content of ΣQNs in water and sediment ranged from 8.0 to 4.4×103 ng·L-1 and 16 to 2.2×103 ng·g-1 in Shijiazhuang water, whereas the primary QNs in water and sediment were enrofloxacin (ENR) and ofloxacin (OFL), respectively. ② The total concentrations of ΣQNs in Shijiazhuang water showed a tendency of being higher in December (1.0×104 ng·L-1) than in April (5.5×103 ng·L-1), and QNs in sediment were also higher in December (7.8×103 ng·g-1) than in April (6.2×103 ng·g-1). ③ The distribution coefficient of QNs in Shijiazhuang water varied from 34 to 2.9×105 L·kg-1 and showed a trend of being greater in December than in April. ④ The results of correlation analysis showed that total nitrogen (TN), nitrate nitrogen (NO3--N), nitrite nitrogen (NO2--N), and ammonia nitrogen (NH4+-N) were significantly correlated with most distribution coefficients of QNs[OFL, norfloxacin (NOR), ENR, difloxacin (DIF), and oxolinic acid (OXO)], whereas temperature (T), total organic carbon (TOC), and total dissolved solids (TDS) were significantly correlated with individual distribution coefficients of QNs[marbofloxacin (MAR) and DIF]. Therefore, the eutrophication level of water affects the distribution behavior of antibiotics in water-sediment.

[Spatial Distribution of Quinolone Antibiotics and Its Correlation Relationship with Microbial Community in Soil of Shijiazhuang City].

Microbial communities are an important component of soil ecosystems. Long-term low content antibiotic pollution will affect the structure and function of microbial communities in soil. Therefore, Shijiazhuang City was selected as the study area, in which twelve sample points were set up in September 2020. These sample sites were divided into four areas (S1, S2, S3, and S4) according to spatial orientation. Ultra-high performance liquid chromatography-mass spectrometry (HPLC-MS/MS) was applied to determine the content of typical antibiotic-quinolones (QNs) in the soil. 16S rRNA high-throughput sequencing technology was used to study the microbial community structure and diversity in the soil. The results showed that:① the total detected contents of QNs in the four areas were S3 (313.5 µg·kg-1)>S4 (65.54 µg·kg-1)>S1 (46.19 µg·kg-1)>S2 (12.63 µg·kg-1). The content of norfloxacin (NOR) was the highest (91.99 µg·kg-1), whereas the content of oxolinic acid (OXO) was the lowest (0.4486 µg·kg-1). ② For grain size, the proportion of powder (2-50 µm) was the highest (66.7%-93.2%), whereas the proportion of clay (less than 2 µm) was the lowest (2.50%-9.10%). For physical and chemical parameters, total phosphorus (TP) and ammonia nitrogen (NH4+-N) showed non-significant spatial differences, whereas nitrate nitrogen (NO3--N), nitrite nitrogen (NO2--N), and grain size showed significant spatial differences. ③ For microbial community composition, there were six dominant bacteria phyla and five dominant bacteria genera, among which Actinobacteriota (18.3%-34.6%) and Proteobacteria (13.6%-34.1%) were the dominant bacteria phyla, and Arthrobacter (3.24%-8.61%) and Nitrososphaeraceae (2.93%-9.46%) were the dominant bacteria genera. The diversity results showed the highest value in the S2 area (6.48) and the lowest value in the S3 area (5.89). ④ QNs and soil physical and chemical parameters significantly changed the structural composition of microbial communities, and OXO, NO3--N, and soil particle size affected the diversity of microbial communities. FLU, NH4+-N, NO2--N, and soil particle size affected the function of the microbial community. Therefore, it is necessary to further strengthen the risk control of antibiotics in the soil of Shijiazhuang City.

[Monitoring and driving force analysis of net primary productivity in native grassland: A case study in Xilingol steppe, China]. / 天然草原净初级生产力变化监测与驱动力分析­­ä»¥é”¡æž—郭勒草原为例.

Grassland is an important type of terrestrial ecosystem. Using remote sensing technology to study the change and driving force of native grassland productivity at large scale is an important way to understand the ecological status of grassland. In this study, potential and actual net primary productivity (NPP) of Xilingol steppe from 2000 to 2018 were examined based on climatic model and light-use efficiency model, respectively. NPP damage value driven by human activities was calculated from the difference between potential and actual NPP. The least square method was used to analyze the temporal and spatial variation of NPP in Xilingol and the driving role of climate and human activities on NPP. The results showed that NPP in Xilingol increased from west to east, with mean annual NPP being 271.54 g C·m-2·a-1, the area with increased NPP (grassland restoration) being 36500 km2, and the area with decreased NPP (grassland degradation) being 59900 km2. The potential NPP tended to rise under the driving force of temperature and precipitation, with an average annual increase of 6.5 g C·m-2·a-1, which indicated that regional climate played a positive role in the improvement of NPP in Xilingol steppe, and that human activities were the main driving force for grassland degradation. The value of NPP damage driven by human activities decreased from east to west and from south to north, with the highest value in Wuzhumuqin meadow and southern steppe. Human activities, such as mining and reclamation, had the most obvious negative impact on grassland NPP.

Protective effect of human umbilical cord mesenchymal stem cell exosomes on preserving the morphology and angiogenesis of placenta in rats with preeclampsia.

AIM: This study aims to investigate the effect of human umbilical cord mesenchymal stem cell exosomes (hucMSC-Ex) on placental tissue and angiogenesis in rats with preeclampsia (PE). METHOD: The expression of MSC surface markers were identified by flow cytometry. Alizarin red staining and oil red O staining were used to examine osteogenic and adipogenic differentiation of hucMSCs. Western blotting was used to determine expressions of CD63 and CD81 in hucMSC-Ex. PE rat models were established using endothelial nitric oxide synthase, eNOS)NG-Nitro-l-arginine Methyl Ester, which were then treated with exosome (Exo) of low dosage (L-Exo), Exo of medium dosage (M-Exo) and Exo of high dosage (H-Exo). The blood pressure at the 15d, 17d and 19d of pregnancy and 24-h urinary protein were measured. TUNEL staining and immunohistochemistry were applied to detect the cell apoptosis and micro-vascular density (MVD) in placental tissues, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to examine serum levels of vascular endothelial growth factor (VEGF) and soluble fms-like tyrosine kinase receptor-1 (sFlt1). RESULTS: In vitro cultured hucMSCs showed expression of MSC surface markers (CD29, CD90 and CD105), and no expression of CD34 and CD45. Besides, the isolated exosomes expressed the exosome markers (CD63 and CD81). In response to the treatment of L-Exo, M-Exo and H-Exo, the blood pressure of PE rat models on the 17 d and the 19 d as well as the 24-h urinary protein were substantially decreased. Moreover, at the 21 d, PE rat models treated with L-Exo, M-Exo and H-Exo exhibited an increase in the number and quality of fetuses, placenta quality, MVD and VEGF expression, but substantial decreased cell apoptosis and expression of sFlt1. The influence of Exos was exerted in a dosage dependent manner. CONCLUSION: hucMSC-Ex, in a dose-dependent manner, can improve the morphology of placental tissue in rats with PE, by inhibiting cell apoptosis and promoting angiogenesis in placental tissue.