Enhancement of carbon nanotubes on microalgal-fungal consortium formation and wastewater treatment.

As a promising material with an efficient light capture capability, a low amount of carbon nanotubes can affect growth and photosynthesis by regulating microalgal cells, thereby enhancing the pollutant removal efficiency in wastewater. In this study, microalgal-fungal consortia of Chlorella vulgaris and Ganoderma lucidum were developed with different types and concentrations of carbon nanotubes. The treatment effect of microalgal-fungal consortia on simulated digestate was also studied. The results demonstrate that 1.5 mg/L of carboxylated multi-walled carbon nanotubes remarkably promoted the formation, growth and photosynthesis of consortium. The dry weight and chlorophyll a content reached 19.3 ± 0.53 mg/symbiont and 27.3 ± 0.52 µg/L, respectively. Moreover, the removal efficiency of chemical oxygen demand, total nitrogen, total phosphorus and tetracycline hydrochloride were 94.1%, 65.5%, 61.9% and 96.2%, respectively. Overall, these findings suggest a promising future for the use of carbon nanotubes in wastewater treatment by regulating microalgal-fungal consortia.

Paeonia lactiflora Pall. Polysaccharide alleviates depression in CUMS mice by inhibiting the NLRP3/ASC/Caspase-1 signaling pathway and affecting the composition of their intestinal flora.

ETHNOPHARMACOLOGICAL RELEVANCE: Paeonia lactiflora Pall. (PL) has been commonly used to de-stressing the liver and relieve depression in traditional Chinese medicine for over a thousand years. Recently, it has been widely used in studies on anti-depressant, anti-inflammatory and regulation of intestinal flora. However, the polysaccharide component has received less attention than the saponin component of PL. AIM OF THE STUDY: This study aimed to elucidate the effects of Paeonia lactiflora polysaccharide (PLP) on depressive behavior in mice in a chronic unpredictable mild stress (CUMS) model and its possible action mechanisms. MATERIALS AND METHODS: A model of chronic depression induced by the CUMS approach. Behavioral experiments were used to assess the success of the CUMS model and the therapeutic impact of PLP. Then the extent of damage to the colonic mucosa was assessed by H&E staining; the extent of neuronal damage was assessed by Nissler staining. Inflammatory factor expression was assessed at different sites in the mouse by enzyme-linked immunoassay (Elisa). The alterations of faecal microflora were detected by 16S rRNA gene sequencing. In the colonic tissues, NLRP3, ASC and Caspase-1 mRNA and protein levels detected by quantitative real-time PCR (qRT-PCR) and Western blot(WB). RUSULTS: PLP can improve depressive behavior in CUMS mice, and colonic mucosal and neuronal damage. Elisa assay showed that PLP could reduce interleukin-1ß (IL-1ß), interleukin-6 (IL-6), tumour necrosis factor-α (TNF-α) levels, and increase 5-Hydroxytryptamine(5-HT) levels in CUMS mice. 16S sequencing analysis showed that PLP could regulate the intestinal flora of CUMS mice and increase their species richness. In addition, PLP significantly inhibited NLRP3/ASC/Caspase-1 signalling pathways activation in the colonic tissues of CUMS mice. CONCLUSIONS: PLP modulates depression-related intestinal ecological dysregulation, increases species richness, and inhibits inflammatory factors levels and NLRP3 inflammasome activation to reduce colonic mucosal and neurons damage, thereby improving depression-like behavior and neurotransmitter release in CUMS mice.

Pharmacodynamic structure of deer antler base protein and its mammary gland hyperplasia inhibition mechanism by mediating Raf-1/MEK/ERK signaling pathway activation.

Mammary gland hyperplasia (MGH) is a common mammary disease whose main pathogenesis is the disruption of estradiol (E2) and progesterone (P) secretion, thereby causing overproliferation of mammary epithelial cells and mammary gland tissue hyperplasia. Deer antler base is a traditional Chinese medicine that has been used for many years to treat MGH. However, its pharmacological mechanism and pharmacodynamic material basis are unclear. In this study, we for the first time used the graded salting method to classify deer antler base protein (CNCP) as CNCP-A, CNCP-B, and CNCP-C and explored the pharmacological mechanism of the anti-MGH properties of CNCP. We found that CNCP could regulate the hormonal levels of E2, P, and follicle stimulating hormone (FSH) and improve the histopathological condition. The potential mechanism might be related to the recombinant C-Raf proto oncogene serine/threonine protein kinase/mitogen-activated protein/extracellular regulated protein kinase (Raf-1/MEK/ERK) signaling pathway. By upregulating the protein expression of the follicle stimulating hormone receptor (FSHR), cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) inhibited the activation of the downstream Raf-1/MEK/ERK signaling pathway, which in turn inhibited the proliferation of mammary epithelial cells. We analyzed the physicochemical properties of CNCP-A, CNCP-B, and CNCP-C and obtained CNCP-C-I by column chromatographic purification of the best pharmacophore protein CNCP. Using high-performance liquid gel filtration chromatography (HPGFC), we determined the molecular weight of CNCP-C-I and identified it by high-performance liquid tandem mass spectrometry (LC-MS/MS) to obtain the first match for a high confidence protein KRT1. This study provides a theoretical basis for the development of effective traditional Chinese medicines with low toxicity levels for the prevention and treatment of mammary gland diseases.

Effect of GR24 concentrations on tetracycline and nutrient removal from biogas slurry by different microalgae-based technologies.

A biogas slurry composed of carbon, nitrogen, phosphorus, and antibiotics was generated. Investigations into the nutrient and tetracycline removal performance of four microalgae-based contaminant removal technologies, including Chlorella vulgaris, C. vulgaris co-cultured with endophytic bacteria, C. vulgaris co-cultured with Ganoderma lucidum, and C. vulgaris co-cultured with G. lucidum and endophytic bacteria, were conducted. The algal-bacterial-fungal consortium with 10-9 M strigolactone (GR24) yielded the maximum growth rate and average daily yield for algae at 0.325 ± 0.03 d-1 and 0.192 ± 0.02 g L-1 d-1, respectively. The highest nutrient/ tetracycline removal efficiencies were 83.28 ± 7.95 % for chemical oxygen demand (COD), 82.62 ± 7.97 % for total nitrogen (TN), 85.15 ± 8.26 % for total phosphorus (TP) and 83.92 ± 7.65 % for tetracycline. Adding an algal-bacterial-fungal consortium with an optimal synthetic analog GR24 concentration is seemingly an encouraging strategy for enhancing pollutant removal by algae, possibly overcoming the challenges of eutrophication and antibiotic pollution.

The mechanism of action of Fangji Huangqi Decoction on epithelial-mesenchymal transition in breast cancer using high-throughput next-generation sequencing and network pharmacology.

ETHNOPHARMACOLOGICAL RELEVANCE: Fangji Huangqi Decoction (FHD) is widely used in traditional Chinese medicine (TCM). FHD has been hypothesized to inhibit the epithelial-mesenchymal transition (EMT) process, which may positively impact breast cancer prevention and treatment. However, its exact mechanism of action is still unknown. AIM OF THE STUDY: This study aimed to screen potential targets of FHD for the treatment of EMT in breast cancer through network pharmacology, and to verify their therapeutic effects in vitro experiments and high-throughput second-generation sequencing. MATERIALS AND METHODS: The data sets of effective components and targets of FHD were established through the Traditional Chinese Medicine Systems Pharmacology database. The GeneCards and OMIM databases were used to establish breast cancer-related target datasets, which were then matched with the TCM target data. The interaction between key target proteins was analyzed using the STRING database; the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases were used to identify the associated biological processes and enriched signal pathways, respectively. The active ingredient disease target network was analyzed using Cytoscape. Finally, next generation sequencing was used to verify the related pathways of FHD intervention in EMT in breast cancer. High-content screening was used to identify the genes/pathways affected by FHD. MDA-MB-231 and HCC-1937 breast cancer cell lines were used to evaluate the impact of FHD on migration, invasion, and EMT. RESULTS: Eighty possible significant targets were identified for the treatment of breast cancer EMT with FHD; GO and KEGG were used to identify 173 cell biological processes associated with breast cancer (P < 0.05), including the NF-κB and PI3K-Akt signaling pathways. The high-throughput sequencing and network pharmacology results were highly consistent. The migration and invasion ability of MDA-MB-231 cells was reduced and their EMT status could be reversed by DSHR2 knockdown. The results of morphology and scratch assays showed that FHD could improve the EMT status of HCC-1973. CONCLUSIONS: This study provides more evidence to support the clinical application of FHD, which has reliable interventional effects on breast cancer EMT. Its therapeutic effects may involve a multi-target, multi-pathway, and multi-mechanism effect.

Contribution of vitamin B12 to biogas upgrading and nutrient removal by different microalgae-based technology.

The algae-based technology has a positive effect on the treatment of biogas slurry and the purification of biogas, while vitamin B12 (VB12) is one of the important regulatory substances in the algae-based cultivation system. In this study, different concentrations of VB12 were used in three microalgal treatment technologies to assess their effect on simultaneous removal of nutrients from biogas slurry and removal of CO2 from raw biogas. Results showed that Chlorella vulgaris exhibited higher growth rate, mean daily productivity, chlorophyll a content, carbonic anhydrase activity and better photosynthetic properties when co-cultivated with Ganoderma lucidum, rather than when co-cultivated with activated sludge or under mono-cultivation. Maximum mean chemical oxygen demand, total nitrogen, total phosphorus and CO2 removal efficiencies were found to be 84.29 ± 8.28%, 83.27 ± 8.14%, 85.27 ± 8.46% and 65.71 ± 6.35%, respectively when microalgae were co-cultivated with Ganoderma lucidum under 100 ng L-1 of VB12. This study shows the potential of microalgae and fungi co-cultivation supplemented with VB12 for the simultaneous upgradation of biogas production as well as for the purification of biogas slurry.

Effects of exogenous GR24 on biogas upgrading and nutrient removal by co-culturing microalgae with fungi under mixed LED light wavelengths.

To realize the synchronous purification of raw biogas and biogas slurry, the algal-fungal symbiont pellets were cultivated by supplementing strigolactone (GR24) under different mixed LED light wavelengths. The optimal light intensity was proved to be red and blue in the ratio of 5:5. The symbionts treated with 10-9 M GR24 had the highest growth rate and mean daily productivity. The extracellular carbonic anhydrase activity and the content of chlorophyll were also affected by GR24 concentrations and mixed light wavelengths. With the induction of 10-9 M GR24, the maximum removal efficiency of chemical oxygen demand, total nitrogen, and total phosphorus reached 76.35 ± 6.87%, 78.77 ± 7.13% and 79.49 ± 7.43%, respectively. Besides, the CO2 removal efficiency reached 59.32 ± 5.19% when the concentration of GR24 was 10-7 M. This work will be beneficial for large-scale biogas slurry purification and biogas upgrading using co-cultivation of microalgae and fungi.

Co-culturing microalgae with endophytic bacteria increases nutrient removal efficiency for biogas purification.

Endophytic bacteria were isolated from Chlorella vulgaris and co-cultured with its host microalgae to determine whether this symbiotic system is suitable for purifying biogas and biogas slurry. Results showed that endophytic bacteria S395-1 and S395-2 belonged to different genera. Both strains promoted microalgae growth while improving photosynthetic performance, carbonic anhydrase activity, nutrient removal efficiency, and CO2 fixation. The optimal bacteria (S395-2)-to-microalgae ratio and co-culture duration were 10:1 and 7 days. Under this condition, the growth rate and carbonic anhydrase activity were 0.196 ± 0.06 d-1 and 31.24 ± 0.28 EU/cell, respectively. The symbiotic system had removal efficiencies of 88.29 ± 5.03%, 88.31 ± 4.29%, 88.21 ± 4.51%, and 68.13 ± 1.69% for chemical oxygen demand, nitrogen, phosphorus, and CO2, respectively. These results will provide a framework for constructing a microalgal-bacterial consortium that can improve wastewater treatment and enhance biogas quality.

Effect of GR24 concentrations on biogas upgrade and nutrient removal by microalgae-based technology.

Freshwater microalgae Chlorella vulgaris was cultured and induced with strigolactone (GR24) to simultaneously eliminate nutrients in biogas slurry and purify biogas. Treatment with 10-7 M GR24 yielded maximum growth rate and mean daily productivity for algae at 0.187 ± 0.06 d-1 and 0.097 ± 0.008 g L-1 d-1, respectively. Results from chlorophyll fluorescence transients method demonstrated that moderate concentration of GR24 could enhance the photosynthetic performance of microalgae. In addition, GR24 affected intracellular carbonic anhydrase activity and chlorophyll-a content. Maximum chemical oxygen demand, total nitrogen, and CO2 removal efficiencies were 78.62 ± 2.36%, 76.47 ± 1.53% and 64.05 ± 1.15% with 10-7 M GR24 induction, respectively. Further, highest total phosphorus removal efficiency (80.27 ± 1.93%) was observed at 10-9 M. The optimal GR24 concentration range was determined to be between 10-9 and 10-7 M in consideration with nutrient and CO2 removal efficiencies.

Effectiveness of various wetland vegetation species on mitigating water pollution from highway runoff.

This study selected and tested five submerged aquatic vegetation-based (SAV) wetlands to improve highway runoff treatment in best management practices. The removal efficiencies of suspended solid (SS), chemical oxygen demand (COD), total nitrogen (TN), ammonia nitrogen ( NH 4 + - N ), and total phosphorus (TP) in the five SAV wetlands were analyzed. Furthermore, the lead (Pb) and zinc (Zn) accumulation capabilities of five submerged macrophytes were determined. The obtained results show that Ceratophyllum demersum wetlands achieved the highest nutrient removal and had the heavy metal accumulation property. Vallisneria natans showed the highest bioaccumulation of Pb among all tested species. Ceratophyllum demersum wetlands showed the highest average removal efficiencies of SS (82.97%), COD (62.08%), TN (77.63%), NH 4 + - N (76.24%), TP (77.55%), Pb (96.24%), and Zn (91.23%). The tendencies of contaminant removal showed seasonal variation, and SAV wetlands performed better in summer than in spring and autumn. Consequently, SAV wetlands showed selectivity for contaminant removal. PRACTITIONER POINTS: Chemical oxygen demand (COD), total nitrogen (TN), and total phosphorus (TP) in highway runoff were removed by submerged aquatic vegetation (SAV). Ceratophyllum demersum and Myriophyllum spicatum wetlands performed well on heavy metal removing. Ceratophyllum demersum showed the highest removal efficiencies of TSS, COD, TN, NH 4 + - N , and TP. The SAV wetlands performed better in summer than in other seasons.

Protocatechualdehyde synergizes with aspirin at the platelet cyclooxygenase-1 level.

Polyphenol-aspirin interactions were recently identified; however, the interaction mode and underlying mechanisms remained elusive. Here, we quantitatively assessed the potential interactions among two important polyphenolic compounds, caffeic acid (CA) and protocatechualdehyde (Pro), and aspirin in the AA-induced platelet aggregation model by applying the isobologram and universal response surface approach (URSA) methods. A molecular docking approach and an originally developed platelet-associated aspirin clearance approach (PAACA) were then applied to explore the potential interaction mechanisms. Although Pro and CA themselves exhibited weak inhibitory effect on arachidonic acid (AA)-induced platelet aggregation and the production of thromboxane B2 (TXB2), both Pro and CA potentiated aspirin action in a synergistic manner. The most prominent synergism was found between Pro and aspirin. Pro formed a stable complex into the cyclooxygenase-1 (COX-1) channel by in silico docking and significantly promoted the platelet-associated aspirin clearance, suggesting that the Pro interaction with COX-1 was favorable to the binding of aspirin with COX-1. Taken together, our findings suggest that the capacity of Pro and potentially other structurally similar polyphenolic compounds on promoting the binding of aspirin on platelet COX-1 might be the main mechanism of their synergism with aspirin.

Mechanism-based pharmacokinetic-pharmacodynamic modeling of the estrogen-like effect of ginsenoside Rb1 on neural 5-HT in ovariectomized mice.

We sought to develop a mechanism-based pharmacokinetic-pharmacodynamic (PK-PD) model to characterize the effects of ginsenoside Rb1 (Rb1) and estradiol (E(2)) on neural 5-hydroxytryptamine (5-HT) concentration in ovariectomized mice. PK data of Rb1 and E(2) were obtained in plasma and brain. Brain levels of 5-HT, tryptophan (TRP), 5-hydroxytryptophan (5-HTP), and 5-hydroxyindoleacetic acid (5-HIAA) were determined after a single intravenous injection of Rb1 (20mg/kg) and E(2) (0.2mg/kg) in ovariectomized mice. The activities of tryptophan hydroxylase (TPH), aromatic amino acid decarboxylase (AAAD), and monoamine oxidase (MAO) were also evaluated. Rb1 and E(2) elevated neural 5-HT levels via TPH activation and MAO inhibition, respectively. Effects were well described by the mechanism-based PK-PD model. The net effect of increased 5-HT induced by MAO inhibition is greater than TPH activation. The increased brain levels of 5-HT induced by Rb1 and E(2) were well described by the present PK-PD model, suggesting the use and further development of this mechanism-based model for the effects of ginsenoside on brain 5-HT levels.

Beneficial estrogen-like effects of ginsenoside Rb1, an active component of Panax ginseng, on neural 5-HT disposition and behavioral tasks in ovariectomized mice.

Decreased 5-hydroxytryptamine (5-HT) concentration in the brain has been linked to central nervous system dysfunctions, especially in menopausal women. Ginsenoside Rb1, a potential phytoestrogen, has been shown to improve central nervous system dysfunctions, comparable to the estrogen treatment. To investigate the estrogen-like effects of ginsenoside Rb1 on neural 5-HT disposition and behavioral tasks, we quantified the concentrations of 5-HT and other related endogenous substances in the frontal cortex and striatum of ovariectomized mice. The activities of tryptophan hydroxylase (TPH), aromatic amino acid decarboxylase (AAAD) and monoamine oxidase (MAO) were also measured to evaluate the synthesis and metabolism of neural 5-HT. Our work shows that both ginsenoside Rb1 and estradiol increased the neural 5-HT concentration. Ginsenoside Rb1 and estradiol administration resulted in elevated TPH and depressed MAO activities, indicating that modulating the synthesis and metabolism of neural 5-HT successfully elevated 5-HT concentration. Ginsenoside Rb1 and estradiol also improved object recognition and decreased immobility time in the forced swimming test. However, a pretreatment with clomiphene (an estrogen receptor antagonist) blocked the beneficial effects of ginsenoside Rb1 and estradiol, suggesting that the estrogen-like effects of ginsenoside Rb1 were estrogen receptor-dependent.

Effects of astragaloside IV on IFN-gamma level and prolonged airway dysfunction in a murine model of chronic asthma.

Astragaloside IV (AST) is the main active constituent of Radix Astragali, a Chinese herb traditionally used to prevent asthma attack from chronic asthma patients. Its efficacy and action mechanisms in asthma attack prevention remain nonetheless to be further explored. In this study, chronic asthma was induced exposing ovalbumin (OVA) sensitized mice to repeated OVA challenges twice every two weeks for 12 weeks. Mice were treated with AST for 4 weeks just after the final challenge. In this murine model of chronic asthma, the airway dysfunction and remodeling remained severe and was accompanied with suppression of the IFN-gamma level in the bronchoalveolar lavage fluid (BALF) even four weeks after the final challenge, indicating that the airway structural changes continued to develop even after interruption of OVA challenges. However, after AST treatment, the airway hyperresponsiveness was sharply relieved, accompanied by the reduction of collagen deposition and mucus production, meanwhile the inflammatory cells were decreased but the IFN-gamma level increased in BALF. In conclusion, AST could prevent the development of chronic asthma, thus reducing asthma attacks. Our results indicated that it should be used as a supplementary therapy on preventing asthma attacks from chronic asthma patients.

Integral pharmacokinetics of multiple lignan components in normal, CCl4-induced hepatic injury and hepatoprotective agents pretreated rats and correlations with hepatic injury biomarkers.

Although pharmacokinetic alternations by hepatic injury have been extensively studied, little is known about the potential influence of hepatoprotective agent's treatment. This study was aimed to investigate the holistic pharmacokinetics of multiple lignans, CYP3A regulations, and their correlations with hepatic injury biomarkers, in hepatic injured rats pretreated with or without schisandra lignan extract (SLE) and dimethyl-diphenyl-bicarboxylate (DDB). Integral pharmacokinetics of multiple lignans based on an AUC-weighting approach was determined in normal, CCl4 induced hepatic injury rats pretreated with or without SLE and DDB. Protein expression and activities of CYP3A were determined. Pharmacokinetic parameters and CYP3A activities were correlated with serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. CCl4 induced acute hepatic injury resulted in a nearly 8-fold enhancement of integral plasma exposures of multiple lignans, which was caused by the significant down-regulation of CYP3A. SLE and DDB pretreatment exhibited potent hepatoprotective effects, accompanied with the restored expression and activity of CYP3A, and the recovery of the respective and integral pharmacokinetics of lignans components. The integral AUC(0-tn) and CYP3A activities correlated well with ALT and AST. This study suggested that the pharmacokinetic regulating effects of hepatoprotective agent's on themselves and co-prescribed drugs should be of concern, and hepatic injury biomarkers may serve as good predictors.