The Bioactive Compounds of Epimedium and Their Potential Mechanism of Action in Treating Osteoporosis: A Network Pharmacology and Experimental Validation Study.

Osteoporosis is a global health challenge characterized by bone loss and microstructure deterioration, which urgently requires the development of safer and more effective treatments due to the significant adverse effects and limitations of existing drugs for long-term treatment. Traditional Chinese medicine, like Epimedium, offers fewer side effects and has been used to treat osteoporosis, yet its active compounds and pharmacological mechanisms remain unclear. In this study, 65 potential active compounds, 258 potential target proteins, and 488 pathways of Epimedium were identified through network pharmacology analysis. Further network analysis and review of the literature identified six potential active compounds and HIF-1α for subsequent experimental validation. In vitro experiments confirmed that 2″-O-RhamnosylIcariside II is the most effective compound among the six potential active compounds. It can promote osteoblast differentiation, bind with HIF-1α, and inhibit both HIF-1α gene and protein expression, as well as enhance COL1A1 protein expression under hypoxic conditions. In vivo experiments demonstrated its ability to improve bone microstructures and reduce bone loss by decreasing bone marrow adipose tissue, enhancing bone formation, and suppressing HIF-1α protein expression. This study is the first to describe the therapeutic effects of 2-O-RhamnosylIcariside II on osteoporosis, which was done, specifically, through a mechanism that targets and inhibits HIF-1α. This study provides a scientific basis for the clinical application of Epimedium and offers a new candidate drug for the treatment of osteoporosis. Additionally, it provides new evidence supporting HIF-1α as a therapeutic target for osteoporosis.

The Bacterial and Fungal Compositions in the Rhizosphere of Asarum heterotropoides Fr. Schmidt var. mandshuricum (Maxim.) Kitag. in a Typical Planting Region.

Asarum is a traditional Chinese medicinal plant, and its dried roots are commonly used as medicinal materials. Research into the traits of the bacteria and fungus in the Asarum rhizosphere and how they relate to the potency of medicinal plants is important. During four cropping years and collecting months, we used ITS rRNA gene amplicon and sequencing to assess the population, diversity, and predominant kinds of bacteria and fungus in the rhizosphere of Asarum. HPLC was used to determine the three bioactive ingredients, namely asarinin, aristolochic acid I, and volatile oil. The mainly secondary metabolites of Asarum, relationships between microbial communities, soil physicochemical parameters, and possible influences on microbial communities owing to various cropping years and collecting months were all statistically examined. The cropping years and collecting months affected the abundance and diversity of rhizosphere bacteria and fungi, but the cropping year had a significant impact on the structures and compositions of the bacterial communities. The rhizosphere microorganisms were influenced by both the soil physicochemical properties and enzyme activities. Additionally, this study revealed that Trichoderma was positively correlated with the three bioactive ingredients of Asarum, while Tausonia showed entirely opposite results. Gibberella and Leptosphaeria demonstrated a significantly negative correlation with asarinin and violate oil, but they were weakly correlated with the aristolochic acid I content. This study revealed variations in the Asarum rhizosphere microorganism population, diversity, and dominant types across four cropping years and collecting months. The relationship between Asarum secondary metabolites, the soil physicochemical properties, enzyme activities, and rhizosphere microorganisms was discussed. Our results will guide the exploration of the soil characteristics and rhizosphere microorganisms' structures by regulating the microbial community to enhance Asarum quality.

Beyond a Spray: Pesticide Application Management in Rural China Based on Quadrilateral Evolutionary Game.

Organic pesticides (OP) produce very little environmental contamination compared with conventional pesticides, but their use is low in rural China. Interest conflicts among participants are analysed for the first time to improve pesticide application management (PAM). Retailers, whose roles are usually little-mentioned, were found to be irreplaceable and so were included in the model as players. A quadrilateral evolutionary game is constructed for PAM and used data from field research in servey representative areas in rural China to estimate the future situation. It demonstrates that OP cannot be chosen by most farmers under the current policy and market environment. The simulation showed that: (i) The probability and economic loss of retailers when providing high-concentration pesticide recommendations positively impact OP application. (ii) The organic certification cost and the successful application probability both benefit environmental recovery in the short term, while the advantages outweigh the disadvantages in the long run. (iii) The cost of strict regulations negatively correlate with OP application; while the purchase price and the corresponding premium provided by intermediates positively/negatively correlates with OP application. This suggests that the environment would be better protected by increasing farmers' pesticide knowledge, reducing retailers' monopoly of influence, and providing subsidies and guidance for organic certification. Moreover, shortening the food supply chain and reducing regulatory costs would also help.

Critical roles of Rad54 in tolerance to apigenin-induced Top1-mediated DNA damage.

Apigenin (APG), a flavone sub-class of flavonoids, possesses a diverse range of biological activities, including anti-cancer and anti-inflammatory effects. Previous studies identified the genotoxicity of APG in certain cancer cells, which may be associated with its anticancer effect. However, the DNA damage repair mechanism induced by APG has remained elusive. In order to clarify the molecular mechanisms, the present study determined the toxicity of APG to the wild-type (WT) DT40 chicken B-lymphocyte cell line, as well as to DT40 cells with deletions in various DNA repair genes, and their sensitivities were compared. It was demonstrated that cells deficient of Rad54, a critical homologous recombination gene, were particularly sensitive to APG. Cell-cycle analysis demonstrated that APG caused an increase in the G2/M-phase population of Rad54- / - cells that was greater than that in WT cells. Furthermore, it was demonstrated by immunofluorescence assay that Rad54- / - cells exhibited significantly increased numbers of γ-phosphorylated H2AX variant histone foci and chromosomal aberrations compared to the WT cells in response to APG. Of note, the in vitro complex of enzyme assay indicated that APG induced increased topoisomerase I (Top1) covalent protein DNA complex in Rad54- / - cells compared to WT cells. Finally, these results were verified using the TK6 human lymphoblastoid cell line and it was demonstrated that, as for DT40 cells, Rad54 deficiency sensitized TK6 cells to APG. The present study demonstrated that Rad54 was involved in the repair of APG-induced DNA damage, which was associated with Top1 inhibition.

Degradation of trimethoprim in aqueous by persulfate activated with nanosecond pulsed gas-liquid discharge plasma.

The persulfate activation by nanosecond pulsed gas-liquid discharge (NPG-LD) is employed to degrade the trimethoprim (TMP) in water. The results show that persulfate addition enhances the degradation of TMP by NPG-LD through an obvious synergetic effect. With treatment time of 50 min, the high removal efficiency and energy yield reach 94.6% and 0.57 gkWh-1 in air NPG-LD with the addition of persulfate, respectively, which is 13.5% and 0.09 gkWh-1 higher than that in solo air NPG-LD, respectively. Correspondingly, the calculated synergetic factor achieves 1.62, indicating the synergetic effect is established. The activation mechanism of persulfate by NPG-LD is analyzed by the measurement of reactive species and the effects of radical scavenger addition on TMP removal. It is found that the synergetic effect between NPG-LD and persulfate is attributed to the increased production of OH, H2O2, and . Besides, the TMP degradation by NPG-LD and persulfate synergetic system is influenced by discharge working gas, pulse voltage, addition dosage of persulfate, initial TMP concentration, and initial pH value. Subsequently, the degradation pathway of TMP is analyzed using LC-MS/MS.

Ultra-high synergetic intensity for humic acid removal by coupling bubble discharge with activated carbon.

Humic acid (HA) removal research focuses on the global water treatment industry. In this work, efficient HA degradation with an ultra-high synergetic intensity is achieved by combined bubble discharge with activated carbon (AC). Adding AC to the discharge greatly improves HA removal efficiency and degradation speed; the synergetic intensity reaches 651.52% in the combined system, and the adsorption residual on AC is 4.52%. After 90 min of treatment, the HA removal efficiency reaches 98.90%, 31.29%, and 7.61% in the plasma-AC combined, solo bubble discharge, and solo AC adsorption systems, respectively. During the plasma process, the number of pore structures and active sites and the amount of oxygen-containing functional groups on the AC surface increase, resulting in a higher adsorption capacity to reactive species (H2O2 and O3) and HA and promoting interactions on the AC surface. For HA mineralization, the presence of AC greatly promotes the destruction of aromatic structures and chromophoric HA functional groups.

Critical roles of tyrosyl-DNA phosphodiesterases in cell tolerance to carnosol-induced DNA damage.

Carnosol is a natural compound with pharmacological action due to its anti-cancer properties. However, the precise mechanism for its anti-carcinogenic effect remains elusive. In this study, we used lymphoblastoid TK6 cell lines to identify the DNA damage and repair mechanisms of carnosol. Our results showed that carnosol induced DNA double-strand breaks (DSBs). We also found that cells lacking tyrosyl-DNA phosphodiesterase 1 (TDP1), an enzyme related to topoisomerase 1 (TOP1), and tyrosyl-DNA phosphodiesterase 2 (TDP2), an enzyme related to topoisomerase 2 (TOP2), were supersensitive to carnosol. Carnosol was found to induce the formation of the TOP1-DNA cleavage complex (TOP1cc) and TOP2-DNA cleavage complex (TOP2cc). When comparing the accumulation of γ-H2AX foci and the number of chromosomal aberrations (CAs) with wild-type (WT) cells, the susceptivity of the TDP1-/- and TDP2-/- cells were associated with an increased DNA damage. Our results provided evidence of carnosol inducing DNA lesions in TK6 cells and demonstrated that the damage induced by carnosol was associated with abnormal topoisomerase activity. We conclude that TDP1 and TDP2 play important roles in the anti-cancer effect of carnosol.

Nonhomologous end joining and homologous recombination involved in luteolin-induced DNA damage in DT40 cells.

Luteolin (3',4',5,7-tetrahydroxyflavone), a naturally occurring flavonoid, has been shown to have anticancer activity in many types of cancer cell lines. The anticancer capacity of luteolin may be related to its ability to induce DNA double-strand breaks (DSBs). Here, we used DT40 cells to determine whether nonhomologous end joining (NHEJ) and homologous recombination (HR) are involved in the repair mechanism of luteolin-induced DNA damage. Cells defective in Ku70 (an enzyme associated with NHEJ) or Rad54 (an enzyme essential for HR) were hypersensitive and presented more apoptosis in response to luteolin. Moreover, the sensitivity and apoptosis of Ku70-/- and Rad54-/- cells were associated with increased DNA damage when the numbers of γ-H2AX foci and chromosomal aberrations (CAs) were compared with those from WT cells. Additionally, after treatment with luteolin, Ku70-/- cells presented more Top2 covalent cleavage complexes (Top2cc). These results indicated that luteolin induced DSBs in DT40 cells and demonstrated that both NHEJ and HR participated in the repair of luteolin-induced DSBs, which might be related to the inhibition of topoisomerases. These results imply that simultaneous inhibition of NHEJ and HR with luteolin treatment would provide a powerful protocol in cancer chemotherapy.

Discharge Regimes Transition and Characteristics Evolution of Nanosecond Pulsed Dielectric Barrier Discharge.

Discharge regime transition in a single pulse can present the breakdown mechanism of nanosecond pulsed dielectric barrier discharge. In this paper, regime transitions between streamer, diffuse, and surface discharges in nanosecond pulsed dielectric barrier discharge are studied experimentally using high resolution temporal-spatial spectra and instantaneous exposure images. After the triggering time of 2-10 ns, discharge was initiated with a stable initial streamer channel propagation. Then, transition of streamer-diffuse modes could be presented at the time of 10-34 ns, and a surface discharge can be formed sequentially on the dielectric plate. In order to analyze the possible reason for the varying discharge regimes in a single discharge pulse, the temporal-spatial distribution of vibrational population of molecular nitrogen N2 (C3Πu, v = 0,1,2) and reduced electric field were calculated by the temporal-spatial emission spectra. It is found that at the initial time, a distorted high reduced electric field was formed near the needle electrode, which excited the initial streamer. With the initial streamer propagating to the dielectric plate, the electric field was rebuilt, which drives the transition from streamer to diffuse, and also the propagation of surface discharge.

Brca1 is involved in tolerance to adefovir dipivoxil­induced DNA damage.

Nucleos(t)ide analogues (NAs) are currently the most important anti­viral treatment option for patients with chronic hepatitis B (CHB). Adefovir dipivoxil (ADV), a diester pro­drug of adefovir, has been widely used for the clinical therapy of hepatitis B virus infection. It has been previously reported that adefovir induced chromosomal aberrations (CAs) in the in vitro human peripheral blood lymphocyte assay, while the genotoxic mechanism remains elusive. To evaluate the possible mechanisms, the genotoxic effects of ADV on the TK6 and DT40 cell lines, as well as DNA repair­deficient variants of DT40 cells, were assessed in the present study. A karyotype assay revealed ADV­induced CAs, particularly chromosomal breaks, in wild­type DT40 and TK6 cells. A γ­H2AX foci formation assay confirmed the presence of DNA damage following treatment with ADV. Furthermore, Brca1­/­ DT40 cells exhibited an increased sensitivity to ADV, while the knockdown of various other DNA damage­associated genes did not markedly affect the sensitivity. These comprehensive genetic studies identified the genotoxic capacity of ADV and suggested that Brca1 may be involved in the tolerance of ADV­induced DNA damage. These results may contribute to the development of novel drugs against CHB with higher therapeutic efficacy and less genotoxicity.

Nanosecond pulsed uniform dielectric barrier discharge in atmospheric air: A brief spectroscopic analysis.

The paper proposes a simple and convenient approach to represent the discharge uniformity of nanosecond-pulse dielectric barrier discharge (DBD) in air by observation of the ratio of N2+ (B3Σu+ → X3Σg+, 0-0, 391.4 nm) to N2 (C3Πu → B3Πg, 2-5, 394.3 nm) intensities. The DBDs at different pulse peak voltages, discharge gap distances, dielectric materials and thicknesses were investigated by recording their single-pulse-shot discharge images and N2+/N2 ratios to verify the feasibility of the above innovative approach. The results show that the ratios of N2+/N2 are in the range of 0.18-0.6within our experimental parameters, which is respect to the reduced electric field (E/N, where E is the field strength and N is gas number density) strength of 260-440 Td (1 Td = 10-17 V·cm2). And it is indicated that a lower N2+/N2 ratio would be found in a higher pulse peak voltage or/and a lower discharge gap distance, which benefits for improving the discharge uniformity of nanosecond-pulse DBD. The thickness and permittivity of dielectric material also affect the E/N strength and discharge uniformity to a certain extent, but the effects are ambiguous due to additional factors of dielectric materials. In addition, the theoretical basis and application scope of this approach were also discussed.

Genistein-induced DNA damage is repaired by nonhomologous end joining and homologous recombination in TK6 cells.

Genistein (GES), a phytoestrogen, has potential chemopreventive and chemotherapeutic effects on cancer. The anticancer mechanism of GES may be related with topoisomerase II associated DNA double-strand breaks (DSBs). However, the precise molecular mechanism remains elusive. Here, we performed genetic analyses using human lymphoblastoid TK6 cell lines to investigate whether non-homologous DNA end joining (NHEJ) and homologous recombination (HR), the two major repair pathways of DSBs, were involved in repairing GES-induced DNA damage. Our results showed that GES induced DSBs in TK6 cells. Cells lacking Ligase4, an NHEJ enzyme, are hypersensitive to GES. Furthermore, the sensitivity of Ligase4-/- cells was associated with enhanced DNA damage when comparing the accumulation of γ-H2AX foci and number of chromosomal aberrations (CAs) with WT cells. In addition, cells lacking Rad54, a HR enzyme, also presented hypersensitivity and increased DNA damages in response to GES. Meanwhile, Treatment of GES-lacking enhanced the accumulation of Rad51, an HR factor, in TK6 cells, especially in Ligase4-/- . These results provided direct evidence that GES induced DSBs in TK6 cells and clarified that both NHEJ and HR were involved in the repair of GES-induced DNA damage, suggesting that GES in combination with inhibition of NHEJ or HR would provide a potential anticancer strategy.

Needle-array to Plate DBD Plasma Using Sine AC and Nanosecond Pulse Excitations for Purpose of Improving Indoor Air Quality.

In this study, needle-array to plate electrode configuration was employed to generate an atmospheric air diffuse discharge using both nanosecond pulse and sine AC voltage as excitation voltage for the purpose of improving indoor air quality. Different types of voltage sources and electrode configurations are employed to optimize electrical field distribution and improve discharge stability. Discharge images, electrical characteristics, optical emission spectra, and plasma gas temperatures in both sine AC discharge and nanosecond pulse discharge were compared and the discharge stability during long operating time were discussed. Compared with the discharge excited by sine AC voltage, the nanosecond pulsed discharge is more homogenous and stable, besides, the plasma gas temperature of nanosecond pulse discharge is much lower. Using packed-bed structure, where γ- Al2O3 pellets are filled in the electrode gap, has obvious efficacy in the production of homogenous discharge. Furthermore, both sine AC discharge and nanosecond pulse discharge were used for removing formaldehyde from flowing air. It shows that nanosecond pulse discharge has a significant advantage in energy cost. And the main physiochemical processes for the generation of active species and the degradation of formaldehyde were discussed.

Spectroscopic and electrical characters of SBD plasma excited by bipolar nanosecond pulse in atmospheric air.

In this paper, an atmospheric surface barrier discharge (SBD) generated by annular electrodes in quartz tube is presented through employing bipolar nanosecond pulse voltage in air. The discharge images, waveforms of pulse voltage and discharge current, and optical emission spectra emitted from the discharges are recorded and calculated. A spectra simulation method is developed to separate the overlap of the secondary diffraction spectra which are produced by grating in monochromator, and N2 (B(3)Πg→A(3)Σu(+)) and O (3p(5)P→3s(5)S2(o)) are extracted. The effects of pulse voltage and discharge power on the emission intensities of OH (A(2)Σ(+)→X(2)Пi), N2(+) (B(2)Σu(+)→X(2)Σg(+)), N2 (C(3)Πu→B(3)Πg), N2 (B(3)Πg→A(3)Σu(+)), and O (3p(5)P→3s(5)S2(o)) are investigated. It is found that increasing the pulse peak voltage can lead to an easier formation of N2(+) (B(2)Σu(+)) than that of N2 (C(3)Πu). Additionally, vibrational and rotational temperatures of the plasma are determined by comparing the experimental and simulated spectra of N2(+) (B(2)Σu(+)→X(2)Σg(+)), and the results show that the vibrational and rotational temperatures are 3250±20K and 350±5K under the pulse peak voltage of 28kV, respectively.