Hispidin Increases Cell Apoptosis and Ferroptosis in Prostate Cancer Cells Through Phosphatidylinositol-3-Kinase and Mitogen-activated Protein Kinase Signaling Pathway.

BACKGROUND/AIM: Chemotherapy is mainly used in the clinical treatment of prostate cancer. Different anticancer mechanisms can induce cell death in various cancers. Reactive oxygen species (ROS) play crucial roles in cell proliferation, differentiation, apoptosis, and signal transduction. It is widely accepted that ROS accumulation is closely related to chemical drug-induced cancer cell death. MATERIALS AND METHODS: We utilized the MTT assay to detect changes in cell proliferation. Additionally, colony formation and wound healing assay were conducted to investigate the effect of hispidin on cell colony formation and migration ability. Fluorescence microscopy was used to detect intracellular and mitochondrial ROS levels, while western blot was used for detection of cell apoptosis. RESULTS: Hispidin treatment significantly decreased viability of PC3 and DU145 cancer cells but exhibited no cytotoxicity in WPMY-1 cells. Furthermore, hispidin treatment inhibited cell migration and colony formation and triggered cellular and mitochondrial ROS accumulation, leading to mitochondrial dysfunction and mitochondrion-dependent apoptosis. Moreover, hispidin treatment induced ferroptosis in PC3 cells. Scavenging of ROS with N-acetyl cysteine significantly inhibited hispidin-induced apoptosis by altering the expression of apoptosis-related proteins, such as cleaved caspase-3, 9, Bax, and Bcl2. Furthermore, hispidin treatment dramatically up-regulated MAPK (involving p38, ERK, and JNK proteins) and NF-kB signaling pathways while down-regulating AKT phosphorylation. Hispidin treatment also inhibited ferroptosis signaling pathways (involving P53, Nrf-2, and HO-1 proteins) in PC3 cells. In addition, inhibiting these signaling pathways via treatment with specific inhibitors significantly reversed hispidin-induced apoptosis, cellular ROS levels, mitochondrial dysfunction, and ferroptosis. CONCLUSION: Hispidin may represent a potential candidate for treating prostate cancer.

Peroxiredoxin I and II as novel therapeutic molecular targets in cervical cancer treatment through regulation of endoplasmic reticulum stress induced by bleomycin.

Cervical cancer, significantly affecting women worldwide, often involves treatment with bleomycin, an anticancer agent targeting breast, ovarian, and cervical cancers by generating reactive oxygen species (ROS) to induce cancer cell death. The Peroxiredoxin (PRDX) family, particularly PRDX1 and 2, plays a vital role in maintaining cellular balance by scavenging ROS, thus mitigating the damaging effects of bleomycin-induced mitochondrial and cellular oxidative stress. This process reduces endoplasmic reticulum (ER) stress and prevents cell apoptosis. However, reducing PRDX1 and 2 levels reverses their protective effect, increasing apoptosis. This research highlights the importance of PRDX1 and 2 in cervical cancer treatments with bleomycin, showing their potential to enhance treatment efficacy by managing ROS and ER stress and suggesting a therapeutic strategy for improving outcomes in cervical cancer treatment.

[Metabarcoding Profiling of Phytoplankton Communities Associated with Algal Blooms and Determining Related Drivers in Baiyangdian Lake].

Phytoplankton are the main cause of algal blooms. To identify bloom algae and assess the risks of the algal blooms in Baiyangdian Lake, a survey on 373 sites was conducted in August 2020. The phytoplankton were studied via both morphological-based density counting and metabarcoding profiling. Then, the bloom degree was classed according to algae density, and the relationship between the community of bloom algae and environmental variables were modeled to determine key factors constraining spatial variation in bloom algae communities. The results showed that more than 95% of the sampling sites were free from the risk of algal blooms(phytoplankton density<2×106 cells·L-1), and only five sites had a slight risk of algal blooms. A total of 90 species with potential of algal blooming were detected, including 20 dominant species, which were mainly affiliated with Chlorophyta, Cyanophyta, and Euglenophyta. Communities of bloom algae significantly varied among different regions(P<0.05). Total phosphorus(TP), total nitrogen(TN), and ammonia nitrogen(NH4+-N) were the key factors significantly affecting the spatial variation in algal bloom communities. At the phylum level, these key factors were significantly positively correlated with Chlorophyta, whereas at the species level, species in Bacillariophyta and Chlorophyta responded significantly to these key factors. Thus, our findings suggested that nutrient levels were significantly related to bloom algae communities, and we proposed that controlling the input of nutrients such as nitrogen and phosphorus and regulating the hydrological process of the lake would be effective management techniques to prevent algal blooms in Baiyangdian Lake.

PRDX1 negatively regulates bleomycin-induced pulmonary fibrosis via inhibiting the epithelial-mesenchymal transition and lung fibroblast proliferation in vitro and in vivo.

BACKGROUND: Pulmonary fibrosis is a major category of end-stage changes in lung diseases, characterized by lung epithelial cell damage, proliferation of fibroblasts, and accumulation of extracellular matrix. Peroxiredoxin 1 (PRDX1), a member of the peroxiredoxin protein family, participates in the regulation of the levels of reactive oxygen species in cells and various other physiological activities, as well as the occurrence and development of diseases by functioning as a chaperonin. METHODS: Experimental methods including MTT assay, morphological observation of fibrosis, wound healing assay, fluorescence microscopy, flow cytometry, ELISA, western blot, transcriptome sequencing, and histopathological analysis were used in this study. RESULTS: PRDX1 knockdown increased ROS levels in lung epithelial cells and promoted epithelial-mesenchymal transition (EMT) through the PI3K/Akt and JNK/Smad signalling pathways. PRDX1 knockout significantly increased TGF-ß secretion, ROS production, and cell migration in primary lung fibroblasts. PRDX1 deficiency also increased cell proliferation, cell cycle circulation, and fibrosis progression through the PI3K/Akt and JNK/Smad signalling pathways. BLM treatment induced more severe pulmonary fibrosis in PRDX1-knockout mice, mainly through the PI3K/Akt and JNK/Smad signalling pathways. CONCLUSIONS: Our findings strongly suggest that PRDX1 is a key molecule in BLM-induced lung fibrosis progression and acts through modulating EMT and lung fibroblast proliferation; therefore, it may be a therapeutic target for the treatment of BLM-induced lung fibrosis.

Peroxiredoxin 5 protects HepG2 cells from ethyl ß-carboline-3-carboxylate-induced cell death via ROS-dependent MAPK signalling pathways.

Peroxiredoxin 5 (PRDX5) is the member of Prxs family, widely reported to be involved in various types of cell death. We previously found that PRDX5 knockdown increases the susceptibility of cell death upon oxidative stress treatment. Ethyl ß-carboline-3-carboxylate (ß-CCE), an alkaloid extracted from Picrasma quassioides, has been reported to play a role in neuronal disease, but its anti-cancer potential on liver cancers remains unknown. Here, we studied the effect of PRDX5 on ethyl ß-carboline-3-carboxylate (ß-CCE)-induced apoptosis of hepatomas. High expression level of PRDX5 was deeply related with the postoperative survival of patients with liver cancer, indicating that PRDX5 may be a biomarker of live cancer processing. Moreover, PRDX5 over-expression in HepG2 cells significantly inhibited ß-CCE-induced cell apoptosis and cellular ROS levels as well as mitochondrial dysfunction. Signalling pathway analysis showed that ß-CCE could significantly up-regulate the ROS-dependent MAPK signalling, which were in turn boosts the mitochondria-dependent cell apoptosis. Moreover, PRDX5 over-expression could reverse the anti-cancer effects induced by ß-CCE in HepG2 cells. Our findings suggest that PRDX5 has a protective role on ß-CCE-induced liver cancer cell death and provides new insights for using its anti-cancer properties for liver cancer treatment.

Acinar Cell-Derived Extracellular Vesicle MiRNA-183-5p Aggravates Acute Pancreatitis by Promoting M1 Macrophage Polarization Through Downregulation of FoxO1.

Acute pancreatitis (AP) is a common cause of a clinically acute abdomen. Crosstalk between acinar cells and leukocytes (especially macrophages) plays an important role in the development of AP. However, the mechanism mediating the interaction between acinar cells and macrophages is still unclear. This study was performed to explore the role of acinar cell extracellular vesicles (EVs) in the crosstalk between acinar cells and macrophages involved in the pathogenesis of AP. EVs derived from caerulein-treated acinar cells induced macrophage infiltration and aggravated pancreatitis in an AP rat model. Further research showed that acinar cell-derived EV miR-183-5p led to M1 macrophage polarization by downregulating forkhead box protein O1 (FoxO1), and a dual-luciferase reporter assay confirmed that FoxO1 was directly inhibited by miR-183-5p. In addition, acinar cell-derived EV miR-183-5p reduced macrophage phagocytosis. Acinar cell-derived EV miR-183-5p promoted the pancreatic infiltration of M1 macrophages and increased local and systemic damage in vivo. Subsequently, miR-183-5p overexpression in macrophages induced acinar cell damage and trypsin activation, thus further exacerbating the disease. In clinical samples, elevated miR-183-5p levels were detected in serum EVs and positively correlated with the severity of AP. EV miR-183-5p might play an important role in the development of AP by facilitating M1 macrophage polarization, providing a new insight into the diagnosis and targeted management of pancreatitis. Graphical abstract of the present study. In our caerulein-induced AP model, miR-183-5p was upregulated in injured acinar cells and transported by EVs to macrophages. miR-183-5p could induce M1 macrophage polarization through downregulation of FoxO1 and the release of inflammatory cytokines, which could aggravate AP-related injuries. Therefore, a vicious cycle might exist between injured ACs and M1 macrophage polarization, which is fulfilled by EV-transported miR-183-5p, leading to sustainable and progressive AP-related injuries.

[Analysis of the Seasonal Changes in Planktonic Microbial Diversity in Urban River Supplied with Reclaimed Water: A Case Study of the North Canal River].

Planktonic microorganisms play an important role in urban aquatic ecosystems; however, environmental changes significantly affect their role in the degradation and transformation of pollutants. The highly artificial North Canal River was chosen as the research area in this study. Seasonal changes in planktonic microbial community structure were studied using 16S rRNA high-throughput sequencing. The seasonal change mechanism of planktonic microbial diversity in urban rivers supplied with reclaimed water and its response relationship with environmental parameters were examined. The results showed that there were significant seasonal changes in the diversity and structure of the planktonic microbial community. The alpha diversity in summer was significantly higher than that in spring, owing to the enhancement of water diffusion capacity caused by seasonal rainfall and physical disturbance of the reclaimed water supply. The beta diversity of the planktonic microbial community in summer was weakened compared to that in spring, also owing to the enhancement of water diffusion capacity. Seasonal runoff and temperature were the main driving factors of the seasonal variation in hydrology and water quality in the highly artificial urban river. The changes in NO2--N and TP caused by seasonal runoff and temperature change were the main reason for planktonic microbial diversity changes in the river. The reductive environment of the river was caused by static and discontinuous flow in the spring. Anaerobic bacteria such as Bacteroidetes related to the degradation of dissolved organic matter and Gracilibacteria related to the denitrification process were dominant in the river. Seasonal runoff and frequent rainfall in summer, as well as the increase in the opening and closing frequency of river sluice gates, enhanced the reoxygenation capacity of the river. This significantly alleviated nutrient pollution in the North River Cannel. Additionally, aerobic bacteria and facultative anaerobic bacteria were dominant species in the river during spring. Cyanobacteria with high temperature characteristics, Chloroflexi and other autotrophic microorganisms, as well as Acidobacteria and Gemmatimonadetes played an important role in the degradation and transformation of pollutants. The results of this study have practical significance for urban river pollution control and ecological restoration with reclaimed water as the recharge water source.

Knockdown of Peroxiredoxin V increased the cytotoxicity of non-thermal plasma-treated culture medium to A549 cells.

Administration of non-thermal plasma therapy via the use of plasma-activated medium (PAM) might be a novel strategy for cancer treatment, as it induces apoptosis by increasing reactive oxygen species (ROS) levels. Peroxiredoxin V (PRDX5) scavenges ROS and reactive nitrogen species and is known to regulate several physiological and pathological reactions. However, its role in lung cancer cells exposed to PAM is unknown. Here, we investigated the effect of PRDX5 in PAM-treated A549 lung cancer cells and determined the mechanism underlying its cytotoxicity. Cell culture medium was treated with low temperature plasma at 16.4 kV for 0, 60, 120, or 180 s to develop PAM. PRDX5 was knocked down in A549 cells via transfection with short hairpin RNA targeting PRDX5. Colony formation and wound healing assays, flow cytometry, fluorescence microscopy, and western blotting were performed to detect the effect of PRDX5 knockdown on PAM-treated A549 cells. PAM showed higher cytotoxicity in lung cancer cells than in control cells, downregulated the mitogen-activated protein kinase signaling pathway, and induced apoptosis. PRDX5 knockdown significantly inhibited cell colony formation and migration, increased ROS accumulation, and reduced mitochondrial membrane potential in lung cancer cells. Hence, PRDX5 knockdown combined with PAM treatment represents an effective option for lung cancer treatment.

Ethyl ß-Carboline-3-Carboxylate Increases Cervical Cancer Cell Apoptosis Through ROS-p38 MAPK Signaling Pathway.

BACKGROUND/AIM: Ethyl ß-carboline-3-carboxylate (ß-CCE) is one of the effective ingredients of Picrasma quassioides (P. quassioides). As a ß-carboline alkaloid, it can antagonize the pharmacological effects of benzodiazepines by regulating neurotransmitter secretion through receptors, thus affecting anxiety and physiology. However, its efficacy in cancer treatment is still unclear. MATERIALS AND METHODS: We explored the effect of b-CCE on SiHa cells using MTT assay, western blot, flow cytometry, LDH release, T-AOC, SOD, and MDA assays. RESULTS: We investigated the cytotoxicity of ß-CCE in SiHa cells and verified that ß-CCE could induce cell apoptosis in a time- and concentration-dependent manner. In this process, treatment with ß-CCE significantly increased the levels of cytoplasmic and mitochondrial reactive oxygen species (ROS), which disturb the oxidation homeostasis by regulating the total antioxidant capacity (T-AOC), superoxide dismutase (SOD) activity, and malondialdehyde (MDA) production. Notably, the addition of N-acetylcysteine (NAC) (ROS scavenger) effectively alleviated ß-CCE-induced apoptosis in SiHa cells. In addition, ß-CCE might activate the p38/MAPK signaling pathway, as the pre-treatment with SB203580 (p38 inhibitor) significantly reduced ß-CCE-induced apoptosis in SiHa cells. CONCLUSION: ß-CCE has an anti-tumor activity. It activates the p38/MAPK signaling pathway by increasing intracellular ROS levels, which subsequently induce SiHa cell apoptosis. Our results provide a novel therapeutic target for treatment of cervical cancer.

Reassessment of the Effect of Transcutaneous Auricular Vagus Nerve Stimulation Using a Novel Burst Paradigm on Cardiac Autonomic Function in Healthy Young Adults.

BACKGROUND: Transcutaneous auricular vagus nerve stimulation (taVNS) may modulate cardiac autonomic function. However, the response rate of the traditional tonic paradigm is low, and the results remain inconsistent. A recent pilot study presented a novel burst paradigm to activate the cardiac parasympathetic system, which might offer a new approach to treat cardiac autonomic function. The present study reassessed the effect of burst taVNS on modulating heart rate variability and explored the difference between burst and traditional tonic paradigms. MATERIALS AND METHODS: Forty-two young adults were recruited for this study. Each participant underwent three types of taVNS with sham (30 sec of stimulation), tonic (25 Hz, 500 µsec), and burst (five pulses at 500 Hz every 200 msec) paradigms, respectively, with simultaneous electrocardiogram recording. One-way analysis of variance, multivariate analysis of variance, and linear regression were used for analysis. Multiple testing was performed using Bonferroni correction. RESULTS: Both burst and tonic paradigms induced a significant decrease in heart rate, which continued until poststimulation, and increased cardiac parasympathetic activity. Moreover, two parasympathetic system indicators showed significant increase only in burst taVNS. The response rates during burst (35.7%) and tonic (38.1%) stimulations were both higher than that during sham stimulation (11.9%). The response to taVNS showed parameter specificity with few nonresponders to the tonic paradigm responding to the burst paradigm. The overall response rate increased from 38.1% in tonic taVNS to 54.8% in taVNS using both burst and tonic paradigms. For both burst and tonic responders, baseline cardiac parasympathetic activity was found to be significantly negatively correlated with changes during stimulation. CONCLUSION: The burst parameter could be used as an alternative strategy for regulating cardiac parasympathetic function by taVNS, which has the potential to be used as a complementary paradigm to traditional tonic taVNS for promoting clinical treatment efficacy.

ATG7-enhanced impaired autophagy exacerbates acute pancreatitis by promoting regulated necrosis via the miR-30b-5p/CAMKII pathway.

The present study was performed to explore whether and how impaired autophagy could modulate calcium/calmodulin-dependent protein kinase II (CAMKII)-regulated necrosis in the pathogenesis of acute pancreatitis (AP). Wistar rats and AR42J cells were used for AP modeling. When indicated, genetic regulation of CAMKII or ATG7 was performed prior to AP induction. AP-related necrotic injury was positively regulated by the incubation level of CAMKII. ATG7 positively modulated the level of CAMKII and necrosis following AP induction, indicating that there might be a connection between impaired autophagy and CAMKII-regulated necrosis in the pathogenesis of AP. microRNA (miR)-30b-5p was predicted and then verified as the upstream regulator of CAMKII mRNA in our setting of AP. Given that the level of miR-30b-5p was negatively correlated with the incubation levels of ATG7 after AP induction, a rescue experiment was performed and indicated that the miR-30b-5p mimic compromised ATG7 overexpression-induced upregulation of CAMKII-regulated necrosis after AP induction. In conclusion, our results indicate that ATG7-enhanced impaired autophagy exacerbates AP by promoting regulated necrosis via the miR-30b-5p/CAMKII pathway.

Survival Benefit from Surgical Resection in Lung Cancer Patients with Brain Metastases: a Single-Center, Propensity-Matched Analysis Cohort Study.

BACKGROUND: Brain metastases (BMs) are the most serious complication of lung cancer, affecting the prognosis of lung cancer patients, and pose distinct clinical challenges. This study was designed to explore the prognostic factors related to lung cancer BM and the value of surgical resection in BMs from lung cancer. METHODS: A retrospective analysis was performed on 714 patients with lung cancer BMs screened between January 2010 and January 2018 at the Sun Yat-sen University Cancer Center. A 1:1 propensity score matching analysis was performed to reduce the potential bias between the surgery and the nonsurgery group. In both the raw and the propensity-score matched dataset, univariate and multivariate Cox proportional hazards regression analyses were used to evaluate risk factors for survival. RESULTS: After matching, 258 patients (129 surgery, 129 no surgery) were analyzed. Multivariate analyses after propensity score matching demonstrated that surgical resection was an independent protective factor for overall survival (OS), and older age, lower Karnofsky Performance Scale (KPS) score, and extracranial metastases were independent risk factors for worse OS. Patients without extracranial metastases, without synchronous BM and with a single BM had a better prognosis. CONCLUSIONS: The findings showed that surgical resection, age, KPS score, and extracranial metastases are independent prognostic factors for predicting the OS of patients with lung cancer BMs, and surgical resection for brain metastatic lesions could significantly improve the OS. However, only certain groups of patients with BMs can benefit from intracranial lesion resection, such as no extracranial metastases and metachronous metastases.

[Analysis on the Spatial Variability Mechanism of the Characteristic Water Quality Factors of Urban River Channel Reclaimed Water].

Due to the limitations of the treatment process of urban sewage treatment plants and the complexity of water sources, the rich inorganic nitrogen and trace persistent organic matter in the reclaimed water cause potential human health risks through lateral leakage or bioaccumulation during the replenishment process of rivers and lakes. Exploring the distribution law of different types of reclaimed water characteristic water quality factors and their formation in reclaimed water replenishment river channels is of great significance to river and lake management. This study takes the Beijing-Tianjin-Hebei section of the North Canal as the research area and explores the spatial variation characteristics of conventional physical parameters, full index, inorganic nitrogen, and salinity hydronium antibiotics in river water quality with the help of clustering, discrimination, principal components, and variance decomposition. The results showed that, although the spatial distribution patterns of different types of water quality factors were consistent, they all showed significant mid-upstream and downstream distribution; however, there were big differences in the degree of variation and the mechanism of variation. The spatial variation of inorganic nitrogen and antibiotics was the most obvious, whereas the variation in conventional physical parameters and the full index was the weakest, and the salinity hydronium showed moderate variation. The spatial variation mechanism of conventional physical parameters was mainly reflected in microbial degradation. The full index was the result of the combined effect of microorganisms, diffusion, the synergy of the two, and a certain degree of source-sink homogeneity. Diffusion was the main mechanism affecting the spatial variation in salinity hydronium. The spatial variation mechanism of inorganic nitrogen was mainly reflected in the source-sink homogenization and microbial degradation; as a secondary mechanism of the spatial variation of inorganic nitrogen, diffusion had a synergistic mode with microbial degradation. Antibiotics, which have great differences in chemical structural stability and biodegradability, showed high spatial variability and had the highest diffusion and microbial synergy mechanism. This research provides a quantitative analysis of the spatial variability mechanism of water quality based on variance decomposition, which has practical guiding significance for the causes of the spatial variability of river pollutants and river management.

[Analysis on Diversity of Plankton Microbial Community in the Beijing-Tianjin-Hebei Section of the North Canal River].

Reclaimed water plays an important role in alleviating the shortage of urban water resources; however, the trace pollutants and pathogens in reclaimed water have an effect on the plankton community in the receiving water. This study investigated the spatial variation mechanism of microbial community diversity in the Beijing-Tianjin-Hebei reach of the Nordkanal River based on the OTUs and phylum level fragment number and fragment abundance data matrix. The results showed that the physical and chemical disturbance caused by the frequent inflow of reclaimed water changed the hydrology and water quality of the water body, and the plankton community could be divided into two different groups along the geographical scale:the medium and upstream clustering (MUC) and the downstream clustering (DC). The analysis of diversity index based on the OTUs data matrix showed that the species diversity of the DC group was significantly higher than that of the MUC group, and the abundance distribution and evenness showed the opposite trend. The species richness was mainly determined by the fragment diversity of the occasional microflora; the evenness was mainly determined by the variation of the abundance of the dominant microflora; the sensitivity of the subcommunity structure with different abundance levels to spatial change was in the order of non-dominant microflora > occasional microflora > dominant microflora. The diversity analysis of the data matrix based on phylum level also showed that the species diversity of the DC group was significantly higher than that of the MUC group, and the change trend of abundance was the opposite; the most sensitive microflora group was the non-dominant phyla, followed by the occasional phyla, and the dominant phyla group was the least sensitive. The data matrix based on the number of level segments of the gate was more sensitive to environmental changes than the multi-degree data matrix based on the level of the gate. The environmental factors significantly related to microbial community were turbidity; permanganate index; oxidation-reduction potential (ORP); macrolide (MLs); tetracycline antibiotic (TCs); and regional response factors of salt ions, carbon, and inorganic nitrogen. In the aspect of abundance and diversity, these phylas that the DC group was significantly more than the MUC group were more significantly negatively correlated with MLs, whereas they were positively correlated with TCs, and these phylas that the MUC group was significantly more than the DC group was more significantly positively correlated with MLs. The research results can provide a theoretical basis and technical guidance for the ecological rehabilitation of urban river courses with reclaimed water as their main water supply source.

Comparative study on the main active components of Baoyuan decoction in plasma and urine of normal and heart failure rats.

The global morbidity and mortality of heart failure has been increasing in recent years. Traditional Chinese medicine (TCM) was increasingly used to treat cardiovascular diseases. Baoyuan decoction (BYD) was a famous classical prescription in China. Modern pharmacological studies showed that it had obvious therapeutic effects on cardiovascular diseases, but its pathological pharmacokinetic studies were unclear. In this research, the absorption of 16 bioactive components in plasma and the excretion of 9 representative components in urine of control rats and isoproterenol (ISO)-induced heart failure rats were studied using the large-volume direct-injection LC-MS method established by our research group. The results indicated that flavonoid constituents exhibited quicker absorption and elimination than saponin constituents after oral administration of BYD. The half-life period of some bioactive compounds in the model group was increased, which contributed to the longer therapeutic effect. The cumulative excretion rate of major flavonoid components of BYD decreased significantly in the ISO-induced heart failure rats.

A Novel System for Evaluating the Inhibition Effect of Drugs on Cytochrome P450 Enzymes in vitro Based on Human-Induced Hepatocytes (hiHeps).

Cytochrome P450 (CYP) is the most important phase I drug-metabolizing enzyme, and the effect of drugs on CYP enzymes can lead to decreased pharmacological efficacy or enhanced toxicity of drugs, but there are many deficiencies in the evaluation models of CYP enzymes in vitro. Human-induced hepatocytes (hiHeps) derived from human fibroblasts by transdifferentiation have mature hepatocyte characteristics. The aim was to establish a novel evaluation system for the effect of drugs on CYP3A4, 1A2, 2B6, 2C9, and 2C19 in vitro based on hiHeps. Curcumin can inhibit many CYP enzymes in vitro, and so the inhibition of curcumin on CYP enzymes was compared by human liver microsomes, human hepatocytes, and hiHeps using UPLC-MS and the cocktail method. The results showed that the IC50 values of CYP enzymes in the hiHeps group were similar to those in the hepatocytes group, which proved the effectiveness and stability of the novel evaluation system in vitro. Subsequently, the evaluation system was applied to study the inhibitory activity of notoginseng total saponins (NS), safflower total flavonoids (SF), and the herb pair of NS-SF on five CYP enzymes. The mechanism of improving efficacy after NS and SF combined based on CYP enzymes was elucidated in vitro. The established evaluation system will become a powerful tool for the research of the effect of drugs on the activity of CYP enzymes in vitro, which has broad application prospects in drug research.

[Contrasting Analysis of Microbial Community Composition in the Water and Sediments of the North Canal Based on 16S rRNA High-Throughput Sequencing].

As the bridge of pollutant exchange between sediments and aquatic ecosystems, microorganisms play an important role in material circulation. However, there are few comparative studies of microorganisms in water and sediment of urban rivers with unconventional water supply, sluice dam, and lining closure. The highly artificial area of Beijing-Tianjin-Hebei section of the North Canal was chosen for this study. We analyze the differences of microbial community composition in water and sediment using high-throughput sequencing. The results show that the microbial communities in the sediments of the North Canal have higher α-diversity than those in the water. With regards to ß-diversity, the similarity of microbial communities in the water is higher than that in the sediment. There is no significant difference in the abundance of Proteobacteria between water and sediments. The abundance of α-Proteobacteria, Actinobacteria, Cyanobacteria, and Verrucomicrobia was higher in water than that of sediment, while the abundance of γ-Proteobacteria, δ-Proteobacteria, Chloroflexi, Firmicutes, and Acidobacteria was higher in sediments than that of water. Aerobic or facultative anaerobes dominated the microbial aquatic system, while anaerobes dominated the sediments. The risk of bacteria releasing pathogens from the sediment into the water habitat is high. The research results provide a scientific basis for revealing the mechanism of microbial community change under river pollution risk in highly artificial reclaimed water.

Identification and characterization of quinoline alkaloids from the root bark of Dictamnus dasycarpus and their metabolites in rat plasma, urine and feces by UPLC/Qtrap-MS and UPLC/Q-TOF-MS.

Quinoline alkaloids are the main bioactive and potentially toxic constituents in the root bark of Dictamnus dasycarpus Turcz. (BXP), a widely used traditional Chinese medicine for the treatment of skin inflammation, eczema and rubella. However, the comprehensive analysis of the chemical components and metabolites of quinoline alkaloids remain unclear. In this study, an integrated strategy by combining UPLC/Q-TOF-MS and UPLC/Qtrap-MS was established to comprehensively profile the quinoline alkaloids from BXP and their metabolites in rat plasma, urine and feces. Q-TOF-MS (MSE mode), Qtrap-MS (EMS, MIM, pMRM and NL mode) were performed for acquiring more precursor ions and clearer precursor product ions. A step-by-step manner based on the diagnostic fragment ions (DFIs), in-house database, ClogP value and dipole moment (µ) was proposed to overcome the complexities due to the similar fragmentation behaviors of the quinoline alkaloids. As a result, a total of 73 quinoline alkaloids were unambiguously or tentatively identified. Among them, 4 furoquinolines, 10 dihydrofuroquinolines, 2 pyranoquinolinones, 4 dihydropyranoquinolinones and 9 quinol-2-ones were characterized in BXP for the first time. Moreover, a total of 98 BXP-related constituents (including 57 prototypes and 41 metabolites) were detected in rat plasma, urine and feces. The metabolic pathways included phase I reactions (O-demethylation, hydroxylation and 2,3-olefinic epoxidation) and phase II reactions (conjugation with glucuronide, sulfate and N-acetylcysteine). In conclusion, the integrated strategy with the proposed stepwise manner is suitable for rapid identifying and characterizing more extensive quinoline alkaloids of BXP in vitro and in vivo. Moreover, the results will be helpful for revealing the pharmacological effective substances or toxic substances of BXP and provide a solid basis for further research.

Evaluation of the effects of notoginseng total saponins (NS), safflower total flavonoids (SF), and the combination of NS and SF (CNS) on the activities of cytochrome P450 enzymes using a cocktail method in rats.

Notoginseng total saponins (NS), safflower total flavonoids (SF), and the combination of NS and SF, namely CNS, are used for the treatment of cardiovascular diseases in clinic. This study developed a cocktail assay involving seven cytochrome P450 (CYP) enzymes to elucidate the effect of NS, SF, and CNS on CYP enzymes and to explore the synergistic effect of CNS in terms of CYP enzymes. Ultra-performance liquid chromatography-MS and reverse-transcription polymerase chain reaction were applied to detect the activities and mRNA expression levels of CYP enzymes. SF exhibited inhibitory effects on CYP1A2, 2B1, 2E1, and 2C11 and induction effects on CYP2C19 and 2D4. NS exhibited induction effects on CYP1A2, 2B1, 2E1, 2C11, 2C19, and 2D4. CNS exhibited induction effects on CYP1A2, 2B1, 2E1, 2C19, and 2D4 and inhibitory effects on CYP3A1 in vivo. Moreover, mRNA expression results were consistent with pharmacokinetic results. Potential herb-drug interactions should be studied closely when SF, NS, or CNS with clinical drugs are metabolized by CYP1A2, 2B1, 2E1, 2C11, 2C19, 2D4, and 3A1. CNS could change the inhibition or induction effects of CYP compared to the NS group, which might be one of the causes for the synergistic effects of the combination of NS and SF.