Low cycle number multiplex PCR: A novel strategy for the construction of amplicon libraries for next-generation sequencing.

Multiplex PCR is a critical step when preparing amplicon library for next-generation sequencing. However, there are several challenges related to multiplex PCR including poor uniformity, nonspecific amplification, and primer-dimers. To address these issues, we propose a novel solution strategy that involves using a low cycle number (<10 cycles) in multiplex PCR and then employing carrier DNAs and magnetic beads for the selection of targeted products. This technique improves the amplicon uniformity while also reducing primer-dimers and PCR artifacts. To evaluate our technique, we initially utilized 120 DNA fragments from mouse genome containing single nucleotide polymorphism (SNP) sites. Sequencing results demonstrated that with only 7 cycles of multiplex PCR, 95.8% of the targeted SNP sites were mapped, with a coverage of at least 1×. The average sequencing depth of all amplicons was 1705.79 ± 1205.30×; 87% of them reached a coverage depth that exceeded 0.2-fold of the average sequencing depth. Our method had a greater uniformity (87%) when compared to Hi-Plex PCR (53.3%). Furthermore, we validated our strategy by randomly selecting 90 primer pairs twice from the initial set of 120 primer-pairs. Next, we used the same protocol to prepare amplicon libraries. The two groups had an average sequencing depth of 1013.30 ± 585.57× and 219.10 ± 158.27×, respectively; over 84% of the amplicons had a sequencing depth that exceeded 0.2-fold of average depth. These results suggest that the use of a low cycle number in multiplex PCR is a cost-effective and efficient approach for the preparation of amplicon libraries.

Highly efficient activation of peroxymonosulfate by ZIF-67 anchored cotton derived for ciprofloxacin degradation.

Metal-organic framework (MOF) and MOF-derived materials have attracted extensive research interest as environmental catalysts. In this study, a composite material (ZIF-67/CCot-8) was successfully prepared using cotton fiber as a substrate and growing ZIF-67 in situ. This material exhibited excellent catalytic performance and significantly improved the efficiency of antibiotics degradation. ZIF-67/CCot-8 at a concentration of 0.05 g/L, combined with 0.2 mM peroxymonosulfate (PMS), removed approximately 97% of ciprofloxacin (CIP) and 99% of tetracycline and sulfamethoxazole within 15 min. The high catalytic efficiency of this catalyst is mainly attributed to the uniform distribution of ZIF-67-derived nanoparticles on the surface of the cotton fibers, providing abundant active sites and thereby significantly enhancing the efficiency of antibiotics degradation. Radical quenching experiments and electron paramagnetic resonance (EPR) analyses revealed that sulfate radicals (SO4•-) and singlet oxygen (1O2) were the main active species. Mass spectrometry (MS) was used to elucidate the CIP degradation pathway. The growth of the roots and stems of soybean sprouts in different water environments (tap water, treated water, and untreated water) was also observed. The results demonstrated a significant improvement in the inhibition of plant growth in the post-degradation CIP solution, indicating a substantial reduction in the toxicity of the degraded aqueous solution. To validate the practicality of the ZIF-67/CCot-8/PMS system, a continuous-flow water-treatment device was designed. This system removed 98% of the CIP solution within 180 min, demonstrating its excellent durability. This study presents a potential pathway for effective antibiotics removal using MOF-derived materials.

Independent and joint associations of monocyte to high-density lipoprotein-cholesterol ratio and body mass index with cardiorenal syndrome: insights from NHANES 2003-2020.

BACKGROUND: With the development of pathophysiology, cardiorenal syndrome (CRS), a complex and severe disease, has received increasing attention. Monocyte to high-density lipoprotein-cholesterol ratio (MHR) and body mass index (BMI) are independent risk factors for cardiovascular diseases, but their association with CRS remains unexplored. This study aims to explore the independent and joint effects of MHR and BMI on CRS. METHODS: We included 42,178 NHANES participants. The determination of CRS referred to the simultaneous presence of cardiovascular disease (identified through self-report) and chronic kidney disease (eGFR < 60 mL/min per 1.73 m²). We employed multivariate weighted logistic regression to evaluate the odds ratio (OR) and 95% confidence interval (CI) for the independent and joint associations of MHR and BMI with CRS. We also conducted restricted cubic spines to explore nonlinear associations. RESULTS: The prevalence of CRS was 3.45% among all participants. An increase in both MHR and BMI is associated with a higher risk of CRS (MHR: OR = 1.799, 95% CI = 1.520-2.129, P < 0.001, P-trend < 0.001; BMI: OR = 1.037, 95% CI = 1.023-1.051, P < 0.001). Individuals who simultaneously fall into the highest quartile of MHR and have a BMI of 30 or more face the highest risk of CRS compared to those in the lowest MHR quartile with a BMI of less than 25 (OR = 3.45, 95% CI = 2.40-4.98, P < 0.001). However, there is no interactive association between MHR and BMI with CRS. CONCLUSIONS: Higher MHR and BMI are associated with higher odds of CRS. MHR and BMI can serve as tools for early prevention and intervention of CRS, respectively.

Combined exposure to manganese and iron decreases oxidative stress-induced nerve damage by increasing Nrf2/HO-1/NQO1 expression.

BACKGROUND: Manganese (Mn) and iron (Fe) are essential trace elements for humans, yet excessive exposure to Mn or Fe can accumulate in the central nervous system (CNS) and cause neurotoxicity. The purpose of this study was to investigate the effects of Mn and Fe exposure, alone or in combination, on inducing oxidative stress-induced neurological damage in rat cortical and SH-SY5Y cells, and to determine whether combined exposure to these metals increases their individual toxicity. METHODS: SH-SY5Y cells and male Sprague-Dawley rats were used to observe the effects of oxidative stress-induced neurological damage induced by exposure to manganese and iron alone or in combination. To detect the expression of anti-oxidative stress-related proteins, Nrf2, HO-1, and NQO1, and the apoptosis-related proteins, Bcl2 and Bax, and the neurological damage-related protein, α-syn. To detect reactive oxygen species generation and apoptosis. To detect the expression of the rat cortical protein Nrf2. To detect the production of proinflammatory cytokines. RESULTS: We demonstrate that juvenile developmental exposure to Mn and Fe and their combination impairs cognitive performance in rats by inducing oxidative stress causing neurodegeneration in the cortex. Mn, Fe, and their combined exposure increased the expression of ROS, Bcl2, Bax, and α-syn, activated the inflammatory factors IL-6 and IL-12, inhibited the activities of SOD and GSH, and induced oxidative stress-induced neurodegeneration both in rats and SH-SY5Y cells. Combined Mn-Fe exposure attenuated the oxidative stress induced by Mn and Fe exposure alone by increasing the expression of antioxidant factors Nrf2, HO-1, and NQO1. CONCLUSION: In both in vivo and in vitro studies, manganese and iron alone or in combination induced oxidative stress, leading to neuronal damage. In contrast, combined exposure to manganese and iron mitigated the oxidative stress induced by exposure to manganese and iron alone by increasing the expression of antioxidant factors. Therefore, studies to elucidate the main causes of toxicity and establish the molecular mechanisms of toxicity should help to develop more effective therapeutic modalities in the future.

Enhanced Light-Harvesting and Energy Transfer in Carbon Dots Embedded Thylakoids for Photonic Hybrid Capacitor Applications.

Nanohybrid photosystems have advantages in converting solar energy into electricity, while natural photosystems based solar-powered energy-storage device is still under developed. Here, we fabricate a new kind of photo-rechargeable zinc-ion hybrid capacitor (ZHC) benefiting from light-harvesting carbon dots (CDs) and natural thylakoids for realizing solar energy harvesting and storage simultaneously. Under solar light irradiation, the embedded CDs in thylakoids (CDs/Thy) can convert the less absorbed green light into highly absorbed red light for thylakoids, besides, Förster resonance energy transfer (FRET) between CDs and Thy also occurs, which facilitates the photoelectrons generation during thylakoids photosynthesis, thereby resulting in 6-fold photocurrent output in CDs/Thy hybrid photosystem, compared to pristine thylakoids. Using CDs/Thy as the photocathode in ZHCs, the photonic hybrid capacitor shows photoelectric conversion and storage features. CDs can improve the photo-charging voltage response of ZHCs to ≈1.2 V with a remarkable capacitance enhancement of 144 % under solar light. This study provides a promising strategy for designing plant-based photonic and electric device for solar energy harvesting and storage.

Environmental oestrogens disrupt testicular descent and damage male reproductive health: Mechanistic insight.

Environmental oestrogens (EEs) as environmental pollutants have been paid much attention due to their impact on congenital malformation of male genitourinary system. Exposure to EEs for prolonged time could hinder testicular descent and cause testicular dysgenesis syndrome. Therefore, it is urgent to understand the mechanisms by which EEs exposure disrupt testicular descent. In this review, we summarize recent advances in our understanding of the process of testicular descent, which is regulated by intricate cellular and molecular networks. Increasing numbers of the components of these networks such as CSL and INSL3 are being identified, highlighting that testicular descent is a highly orchestrated process that is essential to human reproduction and survival. The exposure to EEs would lead to the imbalanced regulation of the networks and cause testicular dysgenesis syndrome such as cryptorchidism, hypospadias, hypogonadism, poor semen quality and testicular cancer. Fortunately, the identification of the components of these networks provides us the opportunity to prevent and treat EEs induced male reproductive dysfunction. The pathways that play an important role in the regulation of testicular descent are promising targets for the treatment of testicular dysgenesis syndrome.

Pyridinic Nitrogen Doped Carbon Dots Supply Electrons to Improve Photosynthesis and Extracellular Electron Transfer of Chlorella pyrenoidosa.

Optimizing photosynthesis is imperative for providing energy and organics for all life on the earth. Here, carbon dots doped with pyridinic nitrogen (named lev-CDs) are synthesized by the one-pot hydrothermal method, and the structure-function relationship between functional groups on lev-CDs and photosynthesis of Chlorella pyrenoidosa (C. pyrenoidosa) is proposed. Pyridinic nitrogen plays a key role in the positive effect on photosynthesis caused by lev-CDs. In detail, lev-CDs act as electron donors to supply photo-induced electrons to P680+ and QA+ , causing electron transfer from lev-CDs to the photosynthetic electron transport chain in the photosystems. In return, the recombination efficiency of electron-hole pairs on lev-CDs decreases. As a result, the electron transfer rate in the electron transport chain, the activity of photosystem II, and the Calvin cycle are enhanced. Moreover, the electron transfer rate between C. pyrenoidosa and external circumstances enhanced by lev-CDs is about 50%, and electrons exported from C. pyrenoidosa can be used to reduce iron(III). This study is of great significance for engineering nanomaterials to improve photosynthesis.

The effect and safety of probiotics on depression: a systematic review and meta-analysis of randomized controlled trials.

PURPOSE: With the escalating social pressures, there has been a continuous rise in the prevalence of depression among the population, leading to substantial healthcare burdens. Moreover, conventional pharmacological interventions still exhibit certain limitations. Therefore, the primary objective of this study is to systematically evaluate the clinical efficacy of probiotics in the treatment of depression. METHODS: Randomized controlled trials of probiotics in treating depressive symptoms were retrieved from Pubmed, Cochrane Library, Web of Science, Wan Fang database, and CNKI between the establishment of the database and March 2022. The primary outcome was Beck's depression rating scale (BDI) scores, while the secondary outcomes were depression scores on the DASS-21 scale, biochemical indicators (IL-6, NO, and TNF-α levels), and adverse events. In addition, Revman 5.3 was used for Meta-analysis and quality evaluation, and Stata 17 was used for the Egger test and Begg's test. A total of 776 patients, including 397 and 379 patients in the experimental and control groups, respectively, were included. RESULTS: The total BDI score of the experimental group was lower than that of the control group (MD = - 1.98, 95%CI - 3.14 to - 0.82), and the score of DASS (MD = 0.90, 95%CI - 1.17 to 2.98), the IL-6 level (SMD = - 0.55, 95%CI - 0.88 to - 0.23), the NO level (MD = 5.27, 95% CI 2.51 to 8.03), and the TNF-α level (SMD = 0.19, 95% CI - 0.25 to 0.63). CONCLUSION: The findings substantiate the therapeutic potential of probiotics in mitigating depressive symptoms by significantly reducing Beck's Depression Inventory (BDI) scores and alleviating the overall manifestation of depression.

Frailty index trajectories in Chinese older adults with diverse levels of social participation: findings from a national population-based longitudinal study.

BACKGROUND: Aging and frailty pose significant challenges globally, placing a substantial burden on healthcare and social services due to their adverse consequences. AIM: The primary objective of this study was to investigate the relationship between social participation and development of frailty transition and trajectory. METHODS: This study utilized data from the CLHLS Cohort, a 10-year follow-up study involving 6713 participants, to investigate the association between social participation and development of frailty. Frailty reflects a comprehensive decline in various body functions. The study employed a group-based trajectory model to analyze the development trajectory of the frailty index and used logistic regression to assess the odds ratio (OR) of frailty risk. RESULTS: We identified two distinct groups of frailty progression trajectories: the "stable development group" and the "rapid growth group." Individuals who engaged in social activities at least once a month, but not daily, exhibited a significant association with an increased risk of transitioning into the "rapid growth group" (OR 1.305, 95% CI 1.032-1.649). Those with social participation less than once a month had an even greater risk (OR 1.872, 95% CI 1.423-2.463). Moreover, low social participation frequency (occasionally/never) has a more pronounced impact on frailty progression in males. CONCLUSION: A higher frequency of social participation is associated with a lower risk of being classified into the "rapid growth group" and a slower rate of frailty index progression. Preventing the progression of frailty can contribute to enhanced support for healthy aging among older adults.

The effects of polyethylene microplastics on the growth, reproduction, metabolic enzymes, and metabolomics of earthworms Eisenia fetida.

The existing data regarding the effects of polyethylene (PE) microplastics (MPs) smaller than 5 mm in size on earthworms are insufficient to fully comprehend their toxicity. In this study, earthworms Eisenia fetida were exposed to artificially added PE at a concentration ranging from 0.05 to 20 g/kg soil (0.005%-2%) for 60 days to determine the concentration range causing negative effects on earthworms and to uncover the potential toxic mechanisms. The individual growth, reproduction, and metabolic enzyme activities, including phase I enzymes (cytochrome P450 [CYP] 1A2, 2B6, 2C9, and 3A4), and phase II metabolic enzymes (superoxide dismutase (SOD), catalase (CAT), and glutathione sulfotransferase (GST)), and metabolomics were measured. The observed variations in responses of multiple cross-scale endpoints indicated that individual indices are less responsive to PE MPs than metabolic enzymes or metabolomics. Despite the absence of significant alterations in growth inhibition based on body weight, PE MPs at concentrations equal to or exceeding 2.5 g/kg were found to exert a toxic effect on earthworms, which was evidenced by significant changes in metabolic enzyme activities (CYP1A2, 2B6, 2C9, and 3A4, SOD, CAT, and GST) and important small molecule metabolites screened based on metabolomics, likely due to the bioaccumulation of PE. The toxicity of PE MPs to earthworms is inferred to be associated with neurotoxicity, oxidative damage, decreased detoxification capacity, energy metabolism imbalance, and impaired amino acid and purine metabolism due to bioaccumulation. The findings of this study will enhance our understanding of the molecular toxicity mechanisms of PE MPs and contribute to a more accurate assessment of the ecological risks posed by PE MPs in soil.

Exploring the impact of water, sanitation and hygiene (WASH), early adequate feeding and access to health care on urban-rural disparities of child malnutrition in China.

To explore the effects of UNICEF-suggested modifiable factors, that is, water, sanitation and hygiene (WASH), early adequate feeding and health care on child malnutrition, and to examine the extent to which each factor contributes to urban-rural disparities of child malnutrition in China. Pooling two waves of regionally representative survey data from Jilin, China, in 2013 and 2018, we report on urban-rural relative risks (RRs) in the prevalence of child stunting, wasting and overweight. We employ Poisson regression to examine the effects of urban-rural setting and the three modifiable factors on the prevalence of each malnutrition outcome, that is, stunting, wasting and overweight. We perform mediation analyses to estimate the extent to which each modifiable factor could explain the urban-rural disparities in each malnutrition outcome. The prevalence of stunting, wasting and overweight were 10.9%, 6.3% and 24.7% in urban, and 27.9%, 8.2% and 35.9% in rural Jilin, respectively. The rural to urban crude RR was 2.55 (95% confidence interval [CI]: 1.92-3.39) for stunting, while the corresponding RRs for wasting and overweight were 1.31 (95% CI: 0.84-2.03) and 1.45 (95% CI: 1.20-1.76), respectively. The rural to urban RR for stunting reduced to 2.01 (95% CI: 1.44-2.79) after adjusting for WASH. The mediation analyses show that WASH could mediate 23.96% (95% CI: 4.34-43.58%) of the urban-rural disparities for stunting, while early adequate feeding and health care had no effects. To close the persistent urban-rural gap in child malnutrition, the specific context of rural China suggests that a multi-sectoral approach is warranted that focuses on the sanitation environment and other wider social determinants of health.

[Research on magneto-acoustic-electrical tomography method based on liquid metal contrast agent and M sequence coded excitation].

Magneto-acoustic-electric tomography (MAET) boasts high resolution in ultrasound imaging and high contrast in electrical impedance imaging, making it of significant research value in the fields of early tumor diagnosis and bioelectrical monitoring. In this study, a method was proposed that combined high conductivity liquid metal and maximum length sequence (M sequence) coded excitation to improve the signal-to-noise ratio. It was shown that, under rotational scanning, the liquid metal significantly improved the signal-to-noise ratio of the inter-tissue magneto-acoustic-electric signal and enhanced the quality of the reconstructed image. The signal-to-noise ratio of the signal was increased by 5.6, 11.1, 21.7, and 45.7 times under the excitation of 7-, 15-, 31-, and 63-bit M sequence code, respectively. The total usage time of 31-bit M sequence coded excitation imaging was shortened by 75.6% compared with single-pulse excitation when the same signal-to-noise ratio was improved. In conclusion, the imaging method combining liquid metal and M-sequence coding excitation has positive significance for improving MAET image quality.

The therapeutic potential of diet on immune-related diseases: based on the regulation on tryptophan metabolism.

Tryptophan (TRP), as an essential amino acid, plays crucial roles in maintaining immune homeostasis due to its complex metabolism pathway, including the microbial metabolism, 5-hydroxytryptamine and kynurenine pathways (KP). Metabolites from these pathways can act antioxidant and endogenous ligand of aryl hydrocarbon receptor (including microbiota metabolites: indole, indole aldehyde, indole acetic acid, indole acrylic acid, indole lactate, indole pyruvate acid, indole propionic acid, skatole, tryptamine, and indoxyl sulfate; and KP metabolites: kynurenine, kynurenic acid, 3-hydroxyanthranilic acid, xanthurenic acid, and cinnabarinic acid) for regulating immune response. In immune-related diseases, the production of pro-inflammatory cytokine activates indoleamine-2,3-dioxygenase, a rate-limiting enzyme of KP, leading to abnormal TRP metabolism in vivo. Many recent studies found that TRP metabolism could be regulated by diet, and the diet regulation on TRP metabolism could therapy related diseases. Accordingly, this review provides a critical overview of the relationships among diet, TRP metabolism and immunity with the aim to seek a treatment opportunity for immune-related diseases.

Effects of multi-walled carbon nanotubes in soil on earthworm growth and reproduction, enzymatic activities, and metabolomics.

Increased production and environmental release of multi-walled carbon nanotubes (MWCNTs) increase soil exposure and potential risk to earthworms. However, MWCNT toxicity to earthworms remains unclear, with some studies identifying negative effects and others negligible effects. In this study, to determine whether exposure to MWCNTs negatively affects earthworms and to elucidate possible mechanisms of toxicity, earthworms were exposed to sublethal soil concentrations of MWCNTs (10, 50, and 100 mg/kg) for 28 days. Earthworm growth and reproduction, activities of cytochrome P450 (CYP) isoforms (1A2, 2C9, and 3A4) and antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), and glutathione-s-transferase (GST)), and metabolomics were determined. Effects of MWCNTs on earthworms depended on exposure concentration. Exposure to MWCNTs did not significantly affect growth and reproduction of individual earthworms. Exposure to 50 mg/kg MWCNTs significantly increased activities of CYP2C9, CYP3A4, SOD, CAT, and GST but clearly reduced levels of L-aspartate, L-asparagine, and glutamine. With exposure to 100 mg/kg MWCNTs, toxic effects on earthworms were observed, with significant inhibition in activities of CYP isoenzymes and SOD, significant reductions in L-aspartate, L-asparagine, glutamine, and tryptophan, and simultaneous accumulations of citrate, isocitrate, fumarate, 2-oxoglutarate, pyruvate, D-galactose, carbamoyl phosphate, formyl anthranilate, hypoxanthine, and xanthine. Results suggest that toxicity of MWCNTs to earthworms is associated with reduced detoxification capacity, excessive oxidative stress, and disturbance of multiple metabolic pathways, including amino acids metabolism, the tricarboxylic acid cycle, pyruvate metabolism, D-galactose metabolism, and purine metabolism. The study provides new insights to better understand and predict the toxicity of MWCNTs in soil.

Temperature sensitivity of SOM decomposition is linked with a K-selected microbial community.

Temperature sensitivity (Q10 ) of soil organic matter (SOM) decomposition is a crucial parameter to predict the fate of soil carbon (C) under global warming. Nonetheless, the response pattern of Q10 to continuous warming and the underlying mechanisms are still under debate, especially considering the complex interactions between Q10 , SOM quality, and soil microorganisms. We examined the Q10 of SOM decomposition across a mean annual temperature (MAT) gradient from -1.9 to 5.1°C in temperate mixed forest ecosystems in parallel with SOM quality and bioavailability, microbial taxonomic composition, and functional genes responsible for organic carbon decomposition. Within this temperature gradient of 7.0°C, the Q10 values increased with MAT, but decreased with SOM bioavailability. The Q10 values increased with the prevalence of K-strategy of soil microbial community, which was characterized by: (i) high ratios of oligotrophic to copiotrophic taxa, (ii) ectomycorrhizal to saprotrophic fungi, (iii) functional genes responsible for degradation of recalcitrant to that of labile C, and (iv) low average 16S rRNA operon copy number. Because the recalcitrant organic matter was mainly utilized by the K-strategists, these findings independently support the carbon quality-temperature theory from the perspective of microbial taxonomic composition and functions. A year-long incubation experiment was performed to determine the response of labile and recalcitrant C pools to warming based on the two-pool model. The decomposition of recalcitrant SOM was more sensitive to increased temperature in southern warm regions, which might attribute to the dominance of K-selected microbial communities. It implies that climate warming would mobilize the larger recalcitrant pools in warm regions, exacerbating the positive feedback between increased MAT and CO2 efflux. This is the first attempt to link temperature sensitivity of SOM decomposition with microbial eco-strategies by incorporating the genetic information and disentangling the complex relationship between Q10 and soil microorganisms.

Controllable Synthesis of Two Isomers 4H-Chromene and 2,8-Dioxabicyclo[3.3.1]nonane Derivatives under Catalyst-Free Conditions.

A one-pot method for the selective synthesis of two isomers 4H-chromene and 2,8-dioxabicyclo[3.3.1]nonane derivatives was developed without a catalyst and using EtOH/H2O (4:1, v/v) as the solvent. The reaction was conducted under mild conditions, with forming multiple chemical bonds in one pot and high atom economy, and only a stoichiometric amount of H2O is produced as the byproduct. Its selectivity was controlled by thermodynamics and kinetics, and the reasons for the transformation of the two structures are also discussed.

The responses of the growth, cytochrome P450 isoenzymes activities and the metabolomics in earthworms to sublethal doses of dichlorvos in soil.

In this paper, earthworms (Eisenia fetida) were exposed to sublethal doses of dichlorvos (spiked concentration of 0.1, 1.0, 10 mg/kg) in soil for 14 days, the metabolomics and activities of cytochrome P450 (CYP) isoenzymes (CYP1A2, CYP2C9 and CYP3A4) of earthworms were analyzed aiming to identify sensitive biomarkers and reveal possible mode of toxic action. The results showed that CYP1A2 and CYP2C9 activity appeared to be more sensitive than CYP3A4 activity in response to dichlorvos, and that metabolic responses based on the metabolomics depended on both of the length of exposure and exposure dose. Malate, ornithine, glucose, inosine, myo-inositol and some amino acids (glutamine, tryptophan, phenylalanine, tyrosine, leucine, histidine, glutamate, lysine) and CYP isozenzymes may be biomarkers to reveal the toxic effect of dichlorvos on earthworms. Compared to controls, when dichlorvos dose reached 1.0 and 10 mg/kg on day 14, glucose and ornithine increased significantly, malate and some amino acids (glutamine, tryptophan, phenylalanine, tyrosine, leucine) decreased significantly, and activities of CYP1A2 and CYP2C9 were inhibited significantly. The current results suggested that 1.0 and 10 mg/kg dichlorvos for 14 days of exposure blocked energy metabolism, disordered Krebs cycle, interfered amino acids metabolism and evoked toxic effects on earthworms.

Evaluation of the combined toxicity of multi-walled carbon nanotubes and cadmium on earthworms in soil using multi-level biomarkers.

The coexistence of multi-walled carbon nanotubes (MWCNTs) with cadmium (Cd) in soil may cause the combined biological effects, but few study reported about their joint toxic effects on earthworms. Therefore, this study investigated the effects of sub-lethal levels of MWCNTs (10, 50, 100 mg/kg) and Cd (2.0, 10 mg/kg) on earthworms Eisenia fetida for 14 days. The changes in multi-level biomarkers of growth inhibition rate, cytochrome P450 isoenzymes (CYP1A2, 2C9 and 3A4), and small molecular metabolites (metabolomics) were determined. The toxic interaction between MWCNTs and Cd was characterized by the combination of the biomarker integration index (BRI), joint effect index concentration addition index (CAI), and the effect concentration addition index (EAI). The results showed that the single MWCNTs exposure caused insignificant change in most biomarkers, while the combined exposure of MWCNTs (50-100 mg/kg) and 10 mg/kg Cd led to significant changes in ten most important metabolites identified by metabolomics and activities of CYP1A2, 2C9, and 3A4. Compared with the toxicity of Cd alone, the combined toxicity of the mixture was significantly reduced. According to the integration of BRI and CAI/EAI, a clearly antagonistic interaction at relatively low effects was observed between MWCNTs and Cd. The responses of multiple biomarkers suggest the toxic action mode of the mixture on earthworms was related to the oxidative injury, and the disruption of amino acid, purine, and pyrimidine metabolism, and the urea cycle.

Immunomodulatory activity of puerarin in RAW264.7 macrophages and cyclophosphamide-induced immunosuppression mice.

CONTEXT: Puerarin, a natural isoflavone extracted from Radix puerariae, is famous for treating various cardiovascular and cerebrovascular diseases. However, little is known about its direct immunomodulatory activity. OBJECTIVE: This study was designed to investigate the in vitro and in vivo immunomodulatory effects of Radix puerariae by using the murine monocyte-macrophage cell line RAW264.7 and immunosuppressed cyclophosphamide-induced mice. METHODS: MTT and neutral red phagocytosis assays were conducted to evaluate the in vitro immunomodulatory activities of puerarin on cell viability and phagocytosis by measuring the proliferation, phagocytic, nitric oxide (NO) ability, and TNF-α production ability of stimulated and lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Immunosuppressed cyclophosphamide-induced mice were used to evaluate the in vivo immunomodulatory activities of puerarin by measuring IL-4 and IFN-γ, the serum half hemolysis value, spleen and thymus index, and proliferation assay for splenic lymphocytes. RESULTS AND DISCUSSION: Results showed that puerarin improves immunomodulatory activity by increasing cell proliferation, cell phagocytosis, and NO secretion in RAW264.7 macrophages and reduces the abnormal immunologic activity by decreasing cell phagocytosis and NO secretion in LPS-stimulated RAW264.7 macrophages. In addition, puerarin enhanced the immunologic activity of cyclophosphamide-induced immunosuppression mice by increasing the secretion of NO, IFN-γ, and IL-4, the serum half hemolysis value (HC50), the spleen and thymus index, and proliferation for splenic lymphocytes. CONCLUSION: Puerarin exhibited an upregulated immunomodulatory effect on RAW264.7 macrophages and immunosuppression mice. In addition, puerarin had a downregulated immunomodulatory effect on RAW264.7 macrophages. The results suggest that puerarin could be a promising immunomodulator to assist in the treatment of tumors.

Substituent-controlled chemoselective synthesis of multi-substituted pyridones via a one-pot three-component cascade reaction.

An efficient and concise one-pot strategy for the synthesis of multisubstituted pyridones via a one-pot three-component cascade reaction catalyzed by Cs2CO3 under solvent-free conditions has been developed. The substituent-controlled chemoselective cycloaddition process involved steps including a Michael addition/ethanol elimination/intermolecular cyclization sequence utilizing anilines, diethyl acetylenedicarboxylate, and diethyl ethoxymethylenemalonate. In doing so, various 2-pyridone and 4-pyridone species (41 examples) could be obtained in good to excellent yields.