[Research progress in preparations of Panax ginseng].

Panax ginseng, known as the "king of herbs", is a highly valued medicinal plant, and its medicinal parts include roots, stems, leaves, flowers, and fruits, among which the roots are the most commonly used. The main active components of this medicinal plant include triterpenoid saponins, polysaccharides, peptides, and volatile oils. The chemical components and active metabolites endow this herb with a variety of pharmacological effects, and thus this herb is used to treat various diseases and play healthcare roles. Currently, a wide range of preparations of P. ginseng have been officially registered and marketed, including tablets, oral liquids, and injections, which demonstrate good clinical efficacy in regulating immunity, adjuvant treatment of tumors, alleviating fatigue, delaying the aging process, improving glucose and lipid metabolism, treating cardiovascular diseases, and relieving inflammation and pain. The production process and quality standards of these drugs are of great significance to ensure their efficacy. According to the theory that Ginseng Radix et Rhizoma can greatly replenish original Qi, tonify the spleen and lung, promote fluid production to quench thirst, tranquilize mind and enrich the intelligence, this paper systematically summarized the clinical application progress of P. ginseng and rela-ted preparations on the market and prospected the further development of preparations from P. ginseng, providing a reference for further exploring the medicinal value and healthcare function of this herb. The above contents, as an important basis for the in-depth development of P. ginseng and related drugs, increase the possibilities for the application of P. ginseng.

Procedurally Targeted Delivery of Antitumor Drugs Using FAPα-Responsive TPGS Dimer-Based Flower-like Polymeric Micelles.

To overcome the intestinal epithelium barrier and achieve a better antitumor effect, the procedurally targeting flower-like nanomicelles for oral delivery of antitumor drugs were designed based on FAPα-responsive TPGS1000 dimer (TPGS-Gly-Pro-TPGS) and L-carnitine linked poly(2-ethyl-2-oxazoline)-b-poly(D, l-lactide) (Car-PEOz-b-PLA). As expected, compared with unmodified polymeric micelles (TT-PMs) composed of TPGS-Gly-Pro-TPGS, L-carnitine conjugated polymeric micelles (CTT-PMs) formed from both TPGS-Gly-Pro-TPGS and Car-PEOz-b-PLA with favorable stability in simulated gastrointestinal fluid and FAPα-dependent release capability exhibited remarkably enhanced cellular uptake and transmembrane transport through OCTN2 mediation confirmed by fluorescence immunoassay, which was intuitively evidenced by stronger fluorescence within epithelial cells, and the basal side of small intestinal epithelium of mice being given intragastric administration of DiI-labeled micelles. The transport of CTT-PMs across the intestinal epithelium in an intact form was mediated by clathrin along the intracellular transport pathway of endosome-lysosome-ER-Golgi apparatus. Furthermore, both the increased uptake by FAPα-positive U87MG cells and unchangeable uptake by FAPα-negative C6 cells for coumarin-6 (C-6)/CTT-PMs compared with C-6/TT-PMs evidenced the targeting ability of CTT-PMs to FAPα-positive tumor cells. Both OCTN2-mediation and FAPα-responsiveness were beneficial for polymeric micelles to improve the delivery and therapeutic efficiency of antitumor agents, which was further supported by the remarkable enhancement in in vivo antitumor efficacy via promoting apoptosis of tumor cells for paclitaxel (PTX)-loaded CTT-PMs (PTX/CTT-PMs) with low toxicity compared with PTX/TT-PMs. Our findings offered an alternative design strategy for procedurally targeted delivery of chemotherapeutics by an oral route.

Investigation on the mechanism of Ginkgo Folium in the treatment of Non-alcoholic Fatty Liver Disease by strategy of network pharmacology and molecular docking.

BACKGROUND: Ginkgo Folium has a favorable effect on non-alcoholic fatty live disease (NAFLD), but its mechanism remains unclear. OBJECTIVE: The aim of this study is to reveal the underlying mechanism of Ginkgo Folium in the treatment of NAFLD. METHODS: Ingredients of Ginkgo Folium and ingredients-related genes were collected from TCMSP database and SwissTargetPrediction website, respectively. Genecards database was used to obtain NAFLD-related genes. Next, the protein-protein interaction network and key ingredients-genes network were constructed via Cytoscape3.7.0. Based on the Metascape website, gene ontology function analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were carried out for key genes. Finally, molecular docking was performed to present the interaction between components and genes using AutoDock Vina 1.1.2. RESULTS: Eighteen active ingredients and 10 target genes were screened from Ginkgo Folium. AKT1, TNF, EGFR, PTGS2, MAPK8, PPAγ, APP, ESR1, HIFα and PPAα were considered as potential therapeutic targets. These target genes were mainly enriched in insulin resistance, HIF-1, adipocytokine and AMPK signaling pathways. Molecular docking results suggested that Ginkgo Folium active ingredients including luteolin-4'-glucoside, sesamin, luteolin, chryseriol, isorhamnetin and laricitrin showed strong binding capacities with AKT1. CONCLUSION: The study showed that multi-components in Ginkgo Folium interacted with AKT1 and regulated AKT-AMPK/HIF pathway to alleviate NAFLD. Our findings provided an essential role and basis for new anti-NAFLD drug discovery and further research on Ginkgo Folium.

Transformation of dissolved organic matter at a full-scale petrochemical wastewater treatment plant.

Transformation of dissolved organic matter (DOM) in petrochemical wastewater (PCW) treatment has rarely been studied. In this work, low- and high-salinity PCW were collected from a treatment plant and the transformations of DOM at molecular level along the treatment processes of both PCW were comparatively investigated. By using Orbitrap MS, the polar DOM constituents were categorized into five molecular classes namely saturated compounds, aliphatics, highly unsaturated and phenolic compounds (Huph), polyphenols and condensed polycyclic aromatics (Cpla). Aliphatics (58.62%) with low molecular weight (150-250 Da) and O/C (0-0.2) were dominant in raw low-salinity PCW; while Huph (65.03%) with O/C at 0.2-0.8 were rich in raw high-salinity PCW. After full-scale treatment, differentiated DOM constituents in both raw PCWs were transformed into aliphatics and Huph with O/C at 0.3-0.5. Anoxic/Oxic treatment of low-salinity system (L-A/O) removed a high fraction of aliphatics (53.05%); while Huph with low O/C (0.1-0.3) (65.68%) in the effluent of L-A/O were further mineralized by ozonation of low-salinity system (L-ozonation). In comparison, anoxic/oxic treatment of high-salinity system (H-A/O) mainly removed unsaturated Huph (34.10%) and aliphatics (30.86%). This resulted in a decrease of dissolved organic carbon as indicated via Spearman correlation. Different from L-ozonation, ozonation of high-salinity system (H-ozonation) degraded aliphatics (26.09%) and Huph (41.85%) with a relatively high O/C (0.2-1.2). After L-A/O and L-ozonation treatments, remaining saturated compounds that were originated from raw low-salinity PCW, were removed by subsequent biological aerated filter. Comparatively, after H-A/O and H-ozonation treatments, residual Huph and aliphatics which were mainly bio-derivates and ozonated intermediates, were further removed by air flotation filter. Hence, DOM transformation of different PCWs along similar treatments varied significantly. This study provides in-depth insights on DOM transformation along a full-scale PCW treatment process.

Mechanisms of Zhenwu decoction for the treatment of renal fibrosis at various stages: What is the role of Corynebacterium?

Many studies demonstrated that Zhenwu decoction (ZWD) is effective in the treatment of kidney fibrosis, whereas the mechanism remains unclear. In this work, a microbiomics-based strategy was used to investigate the mechanism of protective effects of ZWD on kidney fibrosis. Unilateral ureteral obstruction was used to replicate a rat model of renal fibrosis, and rats were divided into prophylactic, early, and progression stages according to the timing of administration. Feces was collected to perform microbiota evaluation by high-throughput 16S DNA sequencing. The results indicated that Corynebacterium, Alistipes, Dorea, and Lactonifactor were highlighted as key targeted flora of ZWD in the treatment of renal fibrosis, and their biological functions were related to inflammation, immunity, and renal excretion. Especially, Corynebacterium presented a significant positive correlation with the concentration of Cys-C, Scr, and BUN. The studies on the changes in inflammatory cytokines (INF-γ, IL-1ß, IL-4, and TNF-α) and immunoglobulin (IgA, IgM, and IgG) confirmed the beneficial effects of ZWD on kidney fibrosis. Therefore, this study confirmed the protective effect of ZWD against renal fibrosis at various disease stages, and its mechanism was associated with re-establishing dysbiosis of the intestinal microbiota, reducing inflammation, as well as regulating immune functions. In particular, Corynebacterium may be a key flora in the treatment of renal fibrosis.

Enhanced Oral Absorption and Liver Distribution of Polymeric Nanoparticles through Traveling the Enterohepatic Circulation Pathways of Bile Acid.

The intestinal epithelium is known to be a main hindrance to oral delivery of nanoparticles. Even though surface ligand modification can enhance cellular uptake of nanoparticles, the "easy entry and hard across" was frequently observed for many active targeting nanoparticles. Here, we fabricated polymeric nanoparticles relayed by bile acid transporters with monomethoxy poly(ethylene glycol)-poly(D,l-lactide) and deoxycholic acid-conjugated poly(2-ethyl-2-oxazoline)-poly(D,l-lactide) based on structural characteristics of intestine epithelium and the absorption characteristics of endogenous substances. As anticipated, deoxycholic acid-modified polymeric nanoparticles featuring good stability in simulated gastrointestinal fluid could notably promote the internalization of their payload by Caco-2 cells through mediation of apical sodium-dependent bile acid transporter (ASBT) and transmembrane transport of the nanoparticles across Caco-2 cell monolayers via relay-guide of ASBT, ileal bile acid-binding protein, and the heteromeric organic solute transporter (OSTα-OSTß) along with multidrug resistance-associated protein 3 (MRP3) evidenced by competitive inhibition and fluorescence immunoassay, which was further visually confirmed by the stronger fluorescence from C6-labeled nanoparticles inside enterocytes and the basal side of the intestinal epithelium of mice. The transcellular transport of deoxycholic acid-modified nanoparticles in an intact form was mediated by caveolin/lipid rafts and clathrin with intracellular trafficking trace of endosome-lysosome-ER-Golgi apparatus and bile acid transport route. Furthermore, the increased uptake by HepG2 cells compared with unmodified nanoparticles evidenced the target ability of deoxycholic acid-modified nanoparticles to the liver, which was further supported by ex vivo imaging of excised major organs of mice. Thus, this study provided a feasible and potential strategy to further enhance transepithelial transport efficiency and liver-targeted ability of nanoparticles by means of the specific enterohepatic circulation pathways of bile acid.

miR-93-5p suppresses ovarian cancer malignancy and negatively regulate CCND2 by binding to its 3'UTR region.

Ovarian cancer is the most fatal gynecological cancer worldwide, yet the fundamental mechanism of malignancy acquisition in ovarian cancer remains unknown. miRNA has been implicated to a variety of diseases, including cancer initiation and progression. Cyclin-D2 (CCND2) is ubiquitously implicated in cancer uncontrol cell proliferation. Bioinformatic research revealed that CCND2 is a candidate gene for miR-93-5p with a binding site in its 3'UTR region in the current study. Using our ovarian cancer sample, we verified that miR-93-5p is negatively correlated with CCND2 mRNA and protein levels. Luciferase report assay revealed miR-93-5p inhibits CCND2 production through binding to the 3'UTR region. The expression of miR-93-5p in ovarian cancer patient samples was then determined, and a survival analysis was performed. Our findings showed that miR-93-5p is downregulated in ovarian cancer and is a favorable predictive factor in ovarian cancer patient. CCK8 assay, wound healing assay and flow cytometry-based cell cycle and apoptotic cell analyses were employed here. We found that miR-93-5p suppresses ovarian cancer cell proliferation and migration while enhances cell death. Our research certified that miR-93-5p reduces ovarian cancer malignancy by targeting CCND2.

Stromal Barrier-Dismantled Nanodrill-Like and Cancer Cell-Targeted pH-Responsive Polymeric Micelles for Further Enhancing the Anticancer Efficacy of Doxorubicin.

Cancer-associated fibroblasts (CAFs) were believed to establish a tight physical barrier and a dense scaffold for tumor cells to make them maintain immunosuppression and drug resistance, strongly hindering nanoparticles to penetrate into the core of tumor tissues and limiting the performance of tumor cell-targeted nanoparticles. Here, we fabricated the substrate Z-Gly-Pro of fibroblast activation protein α (FAPα) and folic acid-codecorated pH-responsive polymeric micelles (dual ligand-modified PEOz-PLA polymeric micelles, DL-PP-PMs) that possessed nanodrill and tumor cell-targeted functions based on Z-Gly-pro-conjugated poly(2-ethyl-2-oxazoline)-poly(D,l-lactide) (ZGP-PEOz-PLA), folic acid (FA)-conjugated PEOz-PLA (FA-PEOz-PLA), and PEOz-PLA for cancer therapy. The micelles with about 40 nm particle size and a narrow distribution exhibited favorable pH-activated endo/lysosome escape induced by their pH responsibility. In addition, the enhancement of in vitro cellular uptake and cytotoxicity to folate receptors or FAPα-positive cells for doxorubicin (DOX)/DL-PP-PMs compared with DOX/PP-PMs evidenced the dual target ability of DOX/DL-PP-PMs, which was further supported by in vivo biodistribution results. As expected, in the human oral epidermal carcinoma (KB) cells xenograft nude mice model, the remarkable enhancement of antitumor efficacy for DOX/DL-PP-PMs with low toxicity was observed compared with DOX/FA-PP-PMs and DOX/ZGP-PP-PMs. The possible mechanism was elucidated to be the dismantling of the stromal barrier by nanodrill-like DOX/DL-PP-PMs via the deletion of CAFs evidenced by the downregulation of α-SMA and inhibition of their functions proved by the decrease in the microvascular density labeled with CD31 and the reduction in the extracellular matrix detected by the collagen content, thereby promoting tumor penetration and enhancing their uptake by tumor cells. The present research offered an alternative approach integrating anticancer and antifibrosis effects in one delivery system to enhance the delivery efficiency and therapeutic efficacy of anticancer drugs.

Integrated metabolomics and 16S rDNA sequencing to investigate the mechanism of immune-enhancing effect of health Tonic oral liquid.

Health Tonic oral liquid (HT) is a popular functional food in China and is used to enhance host immune response. However, its mechanisms of action are still poorly understood. In this work, we combined ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF MS) serum metabolomics with 16S rDNA sequencing to evaluate the effects of HT on metabolomics profiling and microbial community signatures. Short-chain fatty acids (SCFAs) contents in fecal were quantified through gas chromatography-mass spectrometry (GC-MS). Results indicated that HT use leads to a significant increase in IgG, IgM and IgA. Thirty-four metabolites were identified and quantified using metabolomics, most were aromatic amino acids and metabolites involved in glucose metabolism. HT intervention significantly increased the abundance of Alloprevotella, which may contribute to intestinal barrier integrity and inflammatory response inhabitation. Most SCFAs were highly expressed following HT intake. In summary, HT use maintains glucose and lipid metabolism balance, promotes high expressions of beneficial bacteria, and exerts promising immunomodulatory effects.

Combined Lipidomics and Network Pharmacology Study of Protective Effects of Salvia miltiorrhiza against Blood Stasis Syndrome.

Blood stasis syndrome (BSS) is one of the most common symptoms of cardiovascular diseases (CVDs) in traditional Chinese medicine (TCM) theory. Previous studies have identified that Salvia miltiorrhiza (Danshen) has beneficial effects on BSS, but there is no relevant research from the perspective of lipidomics to study the mechanism of Danshen against BSS since hyperlipidemia has been the widely accepted risk factor of CVDs. In this study, lipidomics technology combined with network pharmacology was applied to investigate the pathological mechanism of BSS and the protective effects of Danshen. The lipidomics profiling based on the UPLC-QTOF-MS analysis method was applied to identify the differential metabolites in the plasma of blood stasis rats. The related pathway and potential targets involved in the anti-BSS effects of Danshen were predicted by pathway analysis and network pharmacology. The biochemical results showed that Danshen intervention significantly reduced whole blood viscosity (WBV) at all the shear rates and fibrinogen concentration (FIB) (p < 0.01) and increased activated partial thromboplastin time (APTT) effectively (p < 0.01). We also found that 52 lipid metabolites, including glycerophospholipid, sphingolipid, glycerolipid, plasmalogen, cholesterol ester, and testosterone, were associated with blood stasis. Moreover, Dgka, Hsd17b3, Hsd3b1, Inppl1, Lpl, Pik3ca, Pik3r1, Pla2g1b, Pla2g2a, Soat1, and Soat2 were predicted as potential targets, while glycerophospholipid metabolism, glycerolipid metabolism, steroid and steroid hormone biosynthesis, phosphatidylinositol signaling system, and ether lipid metabolism were involved as shared critical pathways of lipidomics analysis and network pharmacology. Collectively, this study offered a new understanding of the protection mechanism of Danshen against BSS, which provided new insight to explore the protective effects of Danshen.

Influences of rice straw biochar and organic manure on forage soybean nutrient and Cd uptake.

This pot experiment aimed to investigate the influence of rice straw biochar (BC 0, 1, and 3%, w/w) and organic manure (OM 0, 1, and 2%, w/w) addition on the growth, nutrient and cadmium (Cd) uptake of forage soybean in 10 mg Cd kg-1 contaminated soils. Compared with non-biochar treatments, biochar decreased shoot biomass, height and nitrogen (N) contents. Organic manure markedly increased the shoot biomass, shoot phosphorus (P), potassium (K), calcium (Ca) and magnesium (Mg) concentration, and root N, P, Ca contents without biochar addition treatments, while in the case of 3% biochar, there were no significant effects on N, K, Ca, and Mg contents of shoot and root among organic manure treatments. In comparison with other treatments, the minimum Cd content of shoots and roots both occurred in the treatment of BC3%+OM2%, while shoot Cd content reached the maximum value in OM2% treatment. Thus, these results suggested that organic manure addition can elevate forage soybean yield and nutrient content, while biochar had no positive effects. High biochar (3%) addition in combination with highest dose of organic manure (2%) can decline the Cd content of soybean and contribute to the agricultural product safety.

Multi pH-sensitive polymer-drug conjugate mixed micelles for efficient co-delivery of doxorubicin and curcumin to synergistically suppress tumor metastasis.

Combination therapy has been proved to be an effective strategy to inhibit metastasis, however, its efficacy is always compromised by the poor delivery efficiency of drugs. In this study, multi pH-sensitive polymer-drug conjugate mixed micelles were fabricated by the self-assembly of PEOz-PLA-ace-Cur, a conjugate of curcumin (Cur) with poly(2-ethyl-2-oxazoline)-poly(d,l-lactide) (PEOz-PLA) through the linkage of the pH-cleavable acetal bond, and PEOz-PLA-imi-DOX, a conjugate of doxorubicin (DOX) with PEOz-PLA through the linkage of the pH-cleavable benzoic imine bond. The mixed conjugate micelles (PP-Cur/PP-DOX-Mix-PMs) with accurately and conveniently controlled mass ratio of the two drugs were demonstrated to have a small particle size (40-128 nm), high drug loading capacity and pH-dependent drug release behavior. Notably, PP-Cur5/PP-DOX1-Mix-PMs exhibited slower DOX release under physiological conditions compared with PEOz-PLA-imi-DOX micelles, resulting in deeply reduced side effects in vivo. Furthermore, the mixed conjugate micelles showed synergistically enhanced inhibition of MDA-MB-231 cell growth and metastasis evidenced by the results of in vitro anti-invasion, wound healing and anti-migration assessment, and in vivo bioluminescence imaging in nude mice, and significant reduction of the side effects of DOX compared with dual drug physically loaded polymeric micelles. Mechanistic studies demonstrated that the possible inhibitory mechanism of PP-Cur5/PP-DOX1-Mix-PMs on tumor metastasis could be assigned to their inhibition of the invasion, migration, intravasation and extravasation of tumor cells. In conclusion, the multi pH-sensitive polymer-drug conjugate mixed micelles with synergistically enhanced anti-tumor and anti-metastasis activity are potential candidates for safe and effective cancer combination therapy.

Influences of arbuscular mycorrhizae, phosphorus fertiliser and biochar on alfalfa growth, nutrient status and cadmium uptake.

The objective of the study was to explore the influences of arbuscular mycorrhizae (AM), phosphorus (P) fertiliser, biochar application (BC) and their interactions on Medicago sativa growth, nutrient, Cd content and AM fungi-plant symbioses. Applications of both P fertiliser and BC significantly increased total biomass and P and potassium (K) uptake, regardless of AM. When no P fertiliser or BC was used, the shoot biomass and nitrogen (N), P, and K contents in the +AM treatments were 1.39, 1.54, 4.53 and 2.06 times higher than those in the -AM treatments, respectively. AM fungi only elevated the total P uptake by 44.03% when P fertiliser was applied at a rate of 30 mg P kg-1 in the absence of BC addition. With BC application or high-P fertiliser input (100 mg P kg-1), the soil available P was significantly higher than that in the other treatments, and AM fungi significantly reduced the shoot biomass. The minimum Cd concentration occurred in the shoots of alfalfas treated with BC and high-P fertiliser inputs; this concentration was lower than the maximum permitted concentration in China. Although the BC and high-P inputs could eliminate the positive mycorrhizal response, the results suggested that BC application in combination with high-P fertiliser input could not only increase forage yields but also lower Cd concentrations to meet the forage safety standards by the dilution effect.

Growth and elemental uptake of Trifolium repens in response to biochar addition, arbuscular mycorrhizal fungi and phosphorus fertilizer applications in low-Cd-polluted soils.

The aim of this study was to examine the effects of arbuscular mycorrhizal (AM) fungi, biochar (BC) addition and phosphorus (P) fertilizer applications on the mycorrhizal response, biomass and elemental uptake of Trifolium repens in cadmium (Cd)-polluted soils. The results showed that mycorrhizal colonization were significantly decreased by 100 mg P kg-1 fertilizer input. Moreover, AM fungi, BC addition and P fertilizer significantly increased shoot biomass accumulation at all treatments. In the absence of BC, the nitrogen (N), potassium (K), calcium (Ca) and magnesium (Mg) contents in the shoots were not affected by AM fungi after P fertilizer application, but the P content in the shoots significantly increased in response to AM fungi. In the absence of BC, both AM fungi and P fertilizer significantly reduced the Cd concentrations in the plant tissues as well as the soil diethylenetriaminepentaacetic acid (DTPA)-Cd concentration. These results indicated that the translocation factors (TFs) were influenced only by BC addition and that the roots could accumulate greater amounts of Cd than the shoots. On the basis of the hygienic standard for feed in China, the shoot Cd concentration in white clover was below the maximum permitted Cd concentration (1 µg g-1) across all treatments. Therefore, it is suggested that no negative mycorrhizal-white clover symbiotic relationships were observed and T. repens could be a suitable forage species for planting in soils with low concentrations of Cd contamination when BC and P fertilizer are applied.

Biochar shifts biomass and element allocation of legume-grass mixtures in Cd-contaminated soils.

Biochar amendments have been considered to increase the competitive abilities of legumes in mixed cultures. However, little is known about how biochar affects the nutrient and Cd allocation within legume-grass mixtures. Therefore, we conducted a pot experiment to explore the effects of biochar addition rate (0, 1, 2.5, and 5%) on four monocultures, the legume Trifolium repens (Tr), Lolium perenne (Lp), Dactylis glomerata (Dg) and Festuca arundinacea (Fa), and three mixed cultures, i.e., Tr + Lp, Tr + Lp + Dg, and Tr + Lp + Dg + Fa. Regardless of biochar addition, Tr plants showed the lowest aboveground Cd concentration among the monoculture treatments. Compared with non-biochar addition treatment, the 1% biochar addition significantly promoted aboveground biomass accumulation and P, K, Ca, and Mg uptake in the aboveground parts of the Tr monoculture treatments by 39.32, 39.88, 88.27, 69.68, and 51.96%, respectively. Nevertheless, the aboveground biomass and P, K, Ca, and Mg uptake as well as the proportion of these parameters in Tr plants in all plant species mixture treatments decreased after biochar application. Maximum aboveground P and Mg uptake occurred in the four-species mixture treatments without biochar addition, whereas maximum values of these parameters occurred in the three-species mixture treatments with 5% biochar addition. Shoot Cd uptake was not decreased by biochar addition at all plant species treatments. Based on the results, it was suggested that biochar could not reduce Cd uptake by increasing the proportion of legumes in the legume-grass mixtures. The complementarity effects on nutrient uptake in the plant species mixtures depended on the amount of biochar added.

Further Enhancement in Intestinal Absorption of Paclitaxel by Using Transferrin-Modified Paclitaxel Nanocrystals.

The intestinal epithelium is considered to be a major obstacle to the gastrointestinal administration for water-insoluble drugs. To enhance the intestinal absorption of paclitaxel by improving its solubility and overcoming the intestinal epithelium barrier, transferrin-modified paclitaxel nanocrystals were prepared based on the specific transferrin receptor expressed on the apical membrane of the intestinal epithelium and examined to exhibit a mean size of around 178 nm, a rod-like morphology, a sustained release property, and an enhanced in vitro antitumor effect. The in situ intestinal perfusion study proved that the intestinal absorption of transferrin-modified paclitaxel nanocrystals was remarkably enhanced compared with that of Taxol and unmodified paclitaxel nanocrystals, which was further evidenced by the result of pharmacokinetic study. Their transcytosis pathway and intracellular trafficking track were disclosed using Caco-2 cell monolayers. The transcytosis of transferrin-modified paclitaxel nanocrystals and unmodified paclitaxel nanocrystals was principally mediated by clathrin and lipid rafts. The colocalization of both paclitaxel nanocrystals with the organelles observed under confocal microscopy suggested that the late endosomes, lysosomes, ER, and Golgi apparatus played a part in the transcellular transport of both paclitaxel nanocrystals during their transcytosis. Therefore, the designed transferrin-modified drug nanocrystals might have a great potential in the enhancement of intestinal absorption of water-insoluble drugs.

Intestinal oligopeptide transporter PepT1-targeted polymeric micelles for further enhancing the oral absorption of water-insoluble agents.

The intestinal epithelium is the main barrier for nanocarriers to orally deliver poorly water-soluble and absorbed agents. To further improve the transmembrane transport efficiency of polymeric micelles, intestinal oligopeptide transporter PepT1-targeted polymeric micelles were fabricated by Gly-Sar-conjugated poly(ethylene glycol)-poly(d,l-lactic acid). The functionalized polymeric micelles with about 40 nm diameter, uniform spherical morphology and favorable cytocompatibility with Caco-2 cells were demonstrated to distinctly enhance the cellular uptake and transmembrane transport of the loaded agents. The results of intestinal absorption strongly evidenced the higher accumulation of the micelles inside the epithelial cells, at the apical and basolateral sides of the epithelium within the villi in mice. Furthermore, the interaction of Gly-Sar decorated polymeric micelles with PepT1 was explored to promote the internalization of the micelles through fluorescence immunoassay, and the PepT1 level on the membrane of Caco-2 cells treated with the micelles appeared to change in a distinctly time-dependent manner. Both clathrin- and caveolae-mediated pathways were involved in the transcellular transport for undecorated polymeric micelles, while the transcellular transport pathway for Gly-Sar decorated ones was changed to be mainly mediated by clathrin and lipid rafts. The colocalization of Gly-Sar decorated micelles with the organelles observed by confocal laser scanning microscopy indicated that late endosomes, lysosomes, endoplasmic reticulum and Golgi apparatus appeared to participate in the intracellular trafficking progression of the micelles. These results suggested that PepT1-targeted polymeric micelles might have a strong potential to greatly promote the oral absorption of poorly water-soluble and absorbed agents.

Rice straw biochar and phosphorus inputs have more positive effects on the yield and nutrient uptake of Lolium multiflorum than arbuscular mycorrhizal fungi in acidic Cd-contaminated soils.

The purpose of the study was to examine biochar amendment, phosphorus (P) fertilizer and arbuscular mycorrhizal fungi (AMF) on the yield, nutrient and cadmium (Cd) absorption of Lolium multiflorum in acidic soil. It was shown that mycorrhizal inoculation had no positive influence on the plant shoot biomass and the contents of nitrogen (N), P, potassium (K), calcium (Ca) and magnesium (Mg) in plants at all biochar and P level treatments. Irrespective of mycorrhizal inoculation and P level, biochar amendments markedly elevated the soil available P and K uptake in plant tissues. In contrast, biochar significantly decreased the translocation factor of plants, soil exchangeable Cd, and acid and neutral phosphatase activities, regardless of the mycorrhizal inoculation and P fertilizer. Without P fertilization, biochar amendments significantly promoted shoot P content, while biochar amendments significantly reduced shoot P content when P fertilizer was applied. Without biochar application, P fertilizer application significantly promoted the biomass and N uptake of shoots in both AMF inoculation treatments, while P fertilizer increased these only in the presence of biochar and mycorrhizal inoculation. The increased N content induced by the biochar amendment elevated the shoot N:P ratio and alleviated the N deficiency with P fertilizer input. Thus, we concluded that the addition of biochar and P fertilizer showed more positive effects on the promotion of growth and nutrient uptake of L. multiflorum than AMF grown in acidic Cd-contaminated soils.

A novel RON splice variant lacking exon 2 activates the PI3K/AKT pathway via PTEN phosphorylation in colorectal carcinoma cells.

Abnormal expression of the Recepteur d'Origine Nantais (RON) receptor tyrosine kinase is accompanied by the generation of multiple splice or truncated variants, which mediate many critical cellular functions that contribute to tumor progression and metastasis. Here, we report a new RON splice variant in the human colorectal cancer (CRC) cell line HT29. This variant is a 165 kda protein generated by alternative pre-mRNA splicing that eliminates exon 2, causing an in-frame deletion of 63 amino acids in the extracellular domain of the RON ß chain. The deleted transcript was a single chain expressed in the intracellular compartment. Although it lacked tyrosine phosphorylation activity, the RONΔ165E2 variant could phosphorylate phosphatase and tensin homolog (PTEN), thereby activating the PI3K/AKT pathway. In addition, in vitro and in vivo experiments showed that the RONΔ165E2 promoted cell migration and tumor growth. Finally, in an investigation of 67 clinical CRC samples, the variant was highly expressed in about 58% of the samples, and was positively correlated with the invasive depth of the tumor (P < 0.05). These results demonstrate that the novel RONΔ165E2 variant promoted tumor progression while activating the PI3K/AKT pathway via PTEN phosphorylation.