Gubi Zhitong formula alleviates osteoarthritis in vitro and in vivo via regulating BNIP3L-mediated mitophagy.

BACKGROUND: Osteoarthritis (OA) is characterized by degeneration of articular cartilage, leading to joint pain and dysfunction. Gubi Zhitong formula (GBZTF), a traditional Chinese medicine formula, has been used in the clinical treatment of OA for decades, demonstrating definite efficacy. However, its mechanism of action remains unclear, hindering its further application. METHODS: The ingredients of GBZTF were analyzed and performed with liquid chromatography-mass spectrometry (LC-MS). 6 weeks old SD rats were underwent running exercise (25 m/min, 80 min, 0°) to construct OA model with cartilage wear and tear. It was estimated by Micro-CT, Gait Analysis, Histological Stain. RNA-seq technology was performed with OA Rats' cartilage, and primary chondrocytes induced by IL-1ß (mimics OA chondrocytes) were utilized to evaluated and investigated the mechanism of how GBZTF protected OA cartilage from being damaged with some functional experiments. RESULTS: A total of 1006 compounds were identified under positive and negative ion modes by LC-MS. Then, we assessed the function of GBZTF through in vitro and vivo. It was found GBZTF could significantly up-regulate OA rats' limb coordination and weight-bearing capacity, and reduce the surface and sub-chondral bone erosions of OA joints, and protect cartilage from being destroyed by inflammatory factors (iNOS, IL-6, IL-1ß, TNF- α, MMP13, ADAMTS5), and promote OA chondrocytes proliferation and increase the S phage of cell cycle. In terms of mechanism, RNA-seq analysis of cartilage tissues revealed 1,778 and 3,824 differentially expressed genes (DEGs) in model vs control group and GBZTF vs model group, respectively. The mitophagy pathway was most significantly enriched in these DEGs. Further results of subunits of OA chondrocytes confirmed that GBZTF could alleviate OA-associated inflammation and cartilage damage through modulation BCL2 interacting protein 3-like (BNIP3L)-mediated mitophagy. CONCLUSION: The therapeutic effectiveness of GBZTF on OA were first time verified in vivo and vitro through functional experiments and RNA-seq, which provides convincing evidence to support the molecular mechanisms of GBZTF as a promising therapeutic decoction for OA.

Qi-Ju-Di-Huang-Pill delays the progression of diabetic retinopathy.

ETHNOPHARMACOLOGICAL RELEVANCE: Qi-Ju-Di-Huang-Pill (QJDH pill) is a Chinese decoction. Although it is commonly used to treat eye conditions, such as diabetic retinopathy (DR), its exact mechanism of action is unknown. AIM OF THE STUDY: To investigate the specific mechanism by which QJDH pill slows the progression of diabetic retinopathy (DR) based on animal and cellular experiments. MATERIAL AND METHODS: The major components of QJDH pill were characterized by ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLCMS/MS). C57BL/6J mice were randomly divided into five groups as follows: normal group (control group), model group (STZ group), low-dosage QJDH pill group (QJDH-L group), medium-dosage QJDH pill group (QJDH-M group) and high-dosage QJDH pill group (QJDH-H group). Changes in water intake, urination, food intake, and body mass were monitored weekly, while changes in blood glucose were monitored monthly. Fluorescein fundus angiography (FFA), optical coherence tomography angiography (OCTA), and optical coherence tomography (OCT) were utilized to analyze the changes in fundus imaging indications. Hematoxylin & eosin (H&E) and transmission electron microscopy (TEM) were employed to examine histopathologic and ultrastructural changes in retina. The levels of interleukin-6 (IL-6), interleukin-17 (IL-17), tumor necrosis factor-α (TNF-α), and vascular endothelial growth factor (VEGF) in peripheral blood were detected using Enzyme-linked immunosorbent assay (ELISA). The mouse retina apoptotic cells were labeled with green fluorescence via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (Tunel). The protein levels of Bcl-2-Associated X (Bax), B cell lymphoma 2 (Bcl-2), Caspase-3, PI3K, phosphorylated PI3K (p-PI3K), protein kinase B (AKT) and phosphorylated AKT (p-AKT) were quantified by Western blot (WB). The retinal pigment epithelium (RPE) cells were cultured and classified into five groups as follows: normal glucose group (NG group), high glucose group (HG group), high glucose + QJDH pill group (HG + QJDH group), high glucose + inhibitor group (HG + LY294002 group), and high glucose + inhibitor + QJDH pill group (HG + LY294002 + QJDH group). Cell viability and apoptosis were detected via Cell Counting Kit-8 (CCK8) and then analyzed by flow cytometry. RESULTS: In vivo experiments revealed that the QJDH pill effectively reduced blood glucose, symptoms of increased water intake, elevated urination, increased food intake and decreased body mass in DR mice. QJDH pill also slowed the development of a series of fundus imaging signs, such as retinal microangiomas, tortuous dilatation of blood vessels, decreased vascular density, and thinning of retinal thickness, downregulated IL-6, IL-17, TNF-α, and VEGF levels in peripheral blood, and inhibited retinal cell apoptosis by activating the PI3K/AKT signaling pathway. Moreover, in vitro experiments showed that high glucose environment inhibited RPE cell viability and activated RPE cell apoptosis pathway. In contrast, lyophilized powder of QJDH pill increased RPE cell viability, protected RPE cells from high glucose-induced damage, and decreased apoptosis of RPE cells by activating the pi3k pathway. CONCLUSION: QJDH pill induces hypoglycemic, anti-inflammatory effects, anti-VEGF and anti-retinal cell apoptosis by activating PI3K/AKT signaling pathway, and thus can protect the retina and slow the DR progression.

Targeting the PHF8/YY1 axis suppresses cancer cell growth through modulation of ROS.

High levels of mitochondrial reactive oxygen species (mROS) are linked to cancer development, which is tightly controlled by the electron transport chain (ETC). However, the epigenetic mechanisms governing ETC gene transcription to drive mROS production and cancer cell growth remain to be fully characterized. Here, we report that protein demethylase PHF8 is overexpressed in many types of cancers, including colon and lung cancer, and is negatively correlated with ETC gene expression. While it is well known to demethylate histones to activate transcription, PHF8 demethylates transcription factor YY1, functioning as a co-repressor for a large set of nuclear-coded ETC genes to drive mROS production and cancer development. In addition to genetically ablating PHF8, pharmacologically targeting PHF8 with a specific chemical inhibitor, iPHF8, is potent in regulating YY1 methylation, ETC gene transcription, mROS production, and cell growth in colon and lung cancer cells. iPHF8 exhibits potency and safety in suppressing tumor growth in cell-line- and patient-derived xenografts in vivo. Our data uncover a key epigenetic mechanism underlying ETC gene transcriptional regulation, demonstrating that targeting the PHF8/YY1 axis has great potential to treat cancers.

Effects of fermented herbal tea residue on meat quality, rumen fermentation parameters and microbes of black goats.

Herbal tea residue (HTR) is generally considered to be a reusable resource which has still retains considerable proportion of nutrients and active substances. This study aimed to investigate the effects of substitution of whole corn silage with fermented herbal tea residue (FHTR) on meat quality, serum indices, rumen fermentation, and microbes in Chuanzhong black goats. Twenty-two female Chuanzhong black goats (4 months old) with similar weight (9.55 ± 0.95 kg) were selected and randomly divided into two groups. FHTR was used to replace 0% (CON group) and 30% (FHTR group) of whole corn silage in the diets and fed as a total mixed ration (TMR) for Chuanzhong black goats. The adaptation feeding period was 7 days, and the experimental period was 35 days. Results illustrated that the FHTR group had higher value of a* and concentrations of DM and CP and lower rate of water loss (P < 0.05) than the CON group. For the serum indices, goats fed with 30% FHTR had higher (P < 0.05) concentration of CR on day 35. For rumen fermentation, the pH and ratio of acetic acid/propionic acid (AA/PA) in the FHTR group were significantly lower than those in the CON group (P < 0.05). In addition, we studied the goats's rumen microbial community composition and found that the dominant phyla were Firmicutes, Bacteroidetes,and Tenericutes; and the dominant genera were Quinella, Candidatus_Saccharimonas, and Saccharofermentans. There was a significant difference in the beta diversity of the rumen microbiota between groups (P < 0.05). To sum up, the addition of FHTR can affect the meat quality, serum indices, improved rumen fermentation by adjusted the diversity and function of the rumen microbiota.

Application of a three dimensional polyethyleneimine functionalized graphene oxide aerogel as an adsorbent for the determination of phytohormones in ginseng.

Aerogels have attracted considerable attention in sample pretreatment for their outstanding properties, such as the unique porous structure, large surface area and abundant modifiable active sites. The present research reports a three-dimensional interconnected porous network aerogel (PEI-AGO) manufactured based on graphene oxide (GO), polyethyleneimine (PEI) and agar as basic materials through a vacuum freeze-drying treatment. The PEI-AGO aerogel exhibits great potential as a solid phase extraction adsorbent for the selective purification of six endogenous plant hormones in conjunction with high performance liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS). Several factors affecting the extraction efficiency were investigated. Under the optimized extraction conditions, a wide linear range of 0.5-100 ng mL-1 with a good linearity (r > 0.9934) was observed. Low limits of detection (LODs) and limits of quantification (LOQs) were obtained in the range of 0.032-0.155 ng mL-1 and 0.107-0.518 ng mL-1, respectively. Furthermore, the relative recoveries for spiked ginseng samples exhibited remarkable consistency, ranging from 90.2% to 117.6%, with a relative standard deviation (RSD) of ≤9.4% (n = 3). In summary, PEI-AGO has proven to be an effective adsorbent for the pretreatment and enrichment of phytohormones which can be used for the determination of trace endogenous acidic plant hormones in ginseng leaves.

Neferine ameliorates nonalcoholic steatohepatitis through regulating AMPK pathway.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD), peculiarly nonalcoholic steatohepatitis (NASH), has become the main cause of liver transplantation and liver-related death. However, the US Food and Drug Administration has not approved a specific medication for treating NASH. Neferine (NEF), a natural bisbenzylisoquinoline alkaloid separated from the traditional Chinese medicine Nelumbinis plumula, has a variety of pharmacological properties, especially on metabolic diseases. Nevertheless, the anti-NASH effect and mechanisms of NEF remain unclear. PURPOSE: This study aimed to investigate the amelioration of NEF on NASH and the potential mechanisms. STUDY DESIGN: HepG2 cells, hepatic stellate cells (HSCs) and high-fat diet (HFD)+carbon tetrachloride (CCl4) induced C57BL/6 mice were used to observe the effect of NEF against NASH and investigate the engaged mechanism. METHODS: HSCs and HepG2 cells stimulated by oleic acid (OA) were treated with NEF. C57BL/6 mice were fed with HFD+CCl4 to induce NASH mouse model and treated with or without NEF (5 mg/kg or 10 mg/kg, once daily, i.p) for 4 weeks. RESULTS: NEF significantly attenuated the accumulation of lipid droplets, intracellular triglyceride (TG) levels and hepatocytes apoptosis in OA-exposed HepG2 cells. NEF not only enhanced the AMPK and ACC phosphorylation in OA-stimulated HepG2 cells, but also reduced inflammatory response and fibrosis in lipopolysaccharide (LPS)-stimulated HepG2 and in LX-2, respectively. In HFD+CCl4-induced NASH mice, pathological staining confirmed NEF treatment mitigated hepatic lipid deposition, inflammatory cell infiltration as well as hepatic fibrosis. Furthermore, the liver weight, serum and hepatic TG and total cholesterol (TC) and aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were decreased compared with the model group. HFD+CCl4 also induced the upregulation of specific proteins and genes associated to inflammation (ILs, TNF-α, NLRP3, ASC, CCL2 and CXCL10) and hepatic fibrosis (collagens, α-SMA, TGF-ß and TIPM1), which were also suppressed by NEF treatment. CONCLUSION: Our results demonstrated that NEF played a protective role in hepatic steatosis via the regulation of AMPK pathways, which may serve as an attractive candidate for a potential novel strategy on prevention and treatment of NASH.

Effect of oral Xiao-xian decoction combined with acupoint application therapy on pediatric adenoid hypertrophy: A randomized trial.

BACKGROUND: This study aimed to observe the clinical effects of Xiao-xian decoction combined with acupoint application therapy (AAT) for treating pediatric adenoid hypertrophy (AH). METHODS: We randomly divided 93 AH children into 3 groups: AAT alone; Xiao-xian decoction + AAT; control: Montelukast oral therapy. All participants were treated for a month. We used the traditional Chinese medicine syndrome score to evaluate the clinical efficacy and the obstructive sleep apnea-18 scale to evaluate the quality of life. RESULTS: The major symptoms (nasal congestion, open mouth breathing, snoring, and tongue image) and secondary symptoms of patients treated with Xiao-xian decoction + AAT significantly improved compared to before treatment. The pairwise comparison between groups showed that snoring, tongue, secondary symptoms, and total effective rate of the combined treatment group were better than the control and AAT alone. Additionally, the open-mouth breathing, quality of life, and recurrence rate did not differ after treatment. CONCLUSION: Oral Xiao-xian decoction combined with AAT significantly improved the symptoms and signs of nasal congestion, open-mouth breathing, snoring, tongue, and quality of life of AH children and may be used as a long-term treatment for AH.

Sinomenine Hydrochloride Can Ameliorate Benign Prostatic Hyperplasia by Lowering the 5α-Reductase 2 Level and Regulating the Balance between the Proliferation and Apoptosis of Cells.

Benign prostatic hyperplasia (BPH) is a chronic disease that affects the quality of life of older males. Sinomenine hydrochloride (SIN) is the major bioactive alkaloid isolated from the roots of the traditional Chinese medicinal plant Sinomenium acutum Rehderett Wilson. We wondered if the SIN administration exerted a regulatory effect on BPH and its potential mechanism of action. Mice with testosterone propionate-induced BPH subjected to bilateral orchiectomy were employed for in vivo experiments. A human BPH cell line (BPH-1) was employed for in vitro experiments. SIN administration inhibited the proliferation of BPH-1 cells (p < 0.05) by regulating the expression of androgen-related proteins (steroid 5-alpha reductase 2 (SRD5A2), androgen receptors, prostate-specific antigen), apoptosis-related proteins (B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax)) and proliferation-related proteins (proliferating cell nuclear antigen (PCNA), mammalian target of rapamycin, inducible nitric oxide synthase) in vitro. SIN administration decreased the prostate-gland weight coefficient (p < 0.05) and improved the histological status of mice suffering from BPH. The regulatory effects of SIN administration on SRD5A2, an apoptosis-related protein (Bcl-2), and proliferation-related proteins (PCNA, matrix metalloproteinase-2) were consistent with in vitro data. SIN exerted a therapeutic effect against BPH probably related to lowering the SRD5A2 level and regulating the balance between the proliferation and apoptosis of cells. Our results provide an important theoretical basis for the development of plant medicines for BPH therapy.

Composition and antioxidant activity of anthocyanins from Aronia melanocarpa extracted using an ultrasonic-microwave-assisted natural deep eutectic solvent extraction method.

A time-saving, efficient, and environmentally friendly ultrasonic-microwave-assisted natural deep eutectic solvent (UMAE-NADES) extraction method was developed for the extraction of anthocyanins from Aronia melanocarpa. Eight different natural eutectic solvents were screened initially, and choline chloride-glycerol was selected as the extraction solvent. The extraction conditions were optimized using the response surface methodology, and the extraction rate of anthocyanins was higher than those achieved using the traditional ethanol method, natural deep eutectic solvent extraction method, and ultrasonic-microwave-assisted ethanol method. Six anthocyanins, including cyanidin-3-O-galactoside, cyanidin-3-O-glucoside, cyanidin-3-O-arabinoside, cyanidin-3-O-xyloside, cyanidin-3,5-O-dihexoside, and the dimer of cyanidin-hexoside were identified and extracted at a purity of 448.873 mg/g using high performance liquid chromatography-mass spectrometry (HPLC-MS). The compounds extracted using UMAE-NADES had higher antioxidant capacities than those extracted by the other three methods. The UMAE-NADES demonstrated significant efficiency toward the extraction of bioactive substances and has potential utility in the food and pharmaceutical industries.

TMT-Based Quantitative Proteomics Analyses Reveal the Antibacterial Mechanisms of Anthocyanins from Aronia melanocarpa against Escherichia coli O157:H7.

Aronia melanocarpa anthocyanins (AMAs), as natural plant extracts, can control pathogens and are attracting increasing attention. In this study, a tandem mass tag (TMT) quantitative proteomics method combined with multiple reaction monitoring (MRM) was used to explore the antibacterial mechanism of AMAs against Escherichia coli at the protein level. The results showed that 1739 proteins were identified in E. coli treated with AMAs, of which 628 were altered, including 262 downregulated proteins and 366 upregulated proteins. Bioinformatics analysis showed that these differentially expressed proteins have different molecular functions and participate in different molecular pathways. AMAs can affect E. coli protein biosynthesis, DNA replication and repair, oxidative stress response, peptidoglycan biosynthesis, and homeostasis. These pathways induce morphological changes and cell death. The results of this study help understand the molecular mechanism of the inhibitory effect of AMAs on food-borne pathogens and provide a reference for further development of plant-derived antimicrobial agents.

Anti-inflammatory and intestinal microbiota modulation properties of high hydrostatic pressure treated cyanidin-3-glucoside and blueberry pectin complexes on dextran sodium sulfate-induced ulcerative colitis mice.

This study investigated the anti-inflammatory effects of cyanidin-3-glucoside (C3G) and blueberry pectin (BP) complexes on mice with dextran sodium sulfate (DSS)-induced colitis before and after high hydrostatic pressure (HHP) treatment. Real-time polymerase chain reaction (RT-PCR), western blotting, and 16S rDNA sequencing were used to study the expression of inflammation-related factors, activation of signal pathway-related proteins, and changes in the intestinal flora in ulcerative colitis (UC) mice. The results showed that HHP-treated C3G-BP complexes significantly relieved diarrhea and blood loss in the stool of UC mice and alleviated colon shortening. The potential mechanism of action involved reduction in intestinal oxidative stress mRNA expression of pro-inflammatory factors, improvement in anti-inflammatory factor levels, inhibition of the NF-κB signaling pathway, increased protein levels of Bcl-2/Bax and caspase-3/cleaved caspase-3 genes, and improved gut microbiota composition. Compared with other experimental groups, the HHP-treated C3G-BP complexes group exhibited the best anti-inflammatory effect on DSS-induced UC mice. The results may provide new ideas for using C3G-BP complexes for treating UC and help develop better processing methods.

Preparation of Naringenin Nanosuspension and Its Antitussive and Expectorant Effects.

Naringenin (NRG) is a natural flavonoid compound abundantly present in citrus fruits and has the potential to treat respiratory disorders. However, the clinical therapeutic effect of NRG is limited by its low bioavailability due to poor solubility. To enhance the solubility, naringenin nanosuspensions (NRG-NSps) were prepared by applying tocopherol polyethylene glycol succinate (TPGS) as the nanocarrier via the media-milling method. The particle size, morphology, and drug-loading content of NRG-NSps were examined, and the stability was evaluated by detecting particle size changes in different physiological media. NRG-NSps exhibited a flaky appearance with a mean diameter of 216.9 nm, and the drug-loading content was 66.7%. NRG-NSps exhibited good storage stability and media stability. NRG-NSps presented a sustainable release profile, and the cumulative drug-release rate approached approximately 95% within 7 d. NRG-NSps improved the antitussive effect significantly compared with the original NRG, the cough frequency was decreased from 22 to 15 times, and the cough incubation period was prolonged from 85.3 to 121.6 s. Besides, NRG-NSps also enhanced expectorant effects significantly, and phenol red secretion was increased from 1.02 to 1.45 µg/mL. These results indicate that NRG-NSps could enhance the bioavailability of NRG significantly and possess a potential clinical application.

Electrochemical sensing platform based on covalent organic framework materials and gold nanoparticles for high sensitivity determination of theophylline and caffeine.

A new covalent organic framework (COF) has been prepared with 1,3,6,8-tetra(4-formyl phenyl) pyrene (TFPPy) and 2,6-diaminopyridine (DP) as building units through a Schiff base reaction by a simple tube oven heating procedure and the structure of the COF has been characterized in detail. The obtained DP-Py COF is employed to fabricate a novel electrochemical sensing platform for sensitive and selective determination of theophylline (TP) and caffeine (CAF) simultaneously through compounding with AuNPs; the peak positions of TP and CAF are 0.95 V and 1.28 V, respectively. The synergistic effect between DP-Py COF and AuNPs effectively enhances the analytical sensitivity for the target analytes. Under the optimized experimental conditions, the electrochemical sensing platform shows a sensitive voltammetric response and wide linear range to both TP and CAF, and the detection limits are 0.19 µM and 0.076 µM (S/N = 3), respectively. This method has been successfully used for the determination of TP and CAF in compound paracetamol capsules and black tea samples. The recovery and relative standard deviations (RSD) of TP are 99.3~101% and 97.6~101% and 1.3~2.0% and 1.3~2.1%, respectively, and the recovery and RSD of CAF are 96.1~102% and 99.4~104% and 2.8~3.9% and 1.7~3.2%, respectively. Compared with traditional detection methods, the constructed sensing platform has better performance and is expected to be widely used also in other real sample analyses.

Isoorientin exerts a urate-lowering effect through inhibition of xanthine oxidase and regulation of the TLR4-NLRP3 inflammasome signaling pathway.

Isoorientin (ISO), a natural flavonoid compound, has been identified in several plants and its biological activity is determined and the study on lowering uric acid has not been reported. In view of the current status of treatment of hyperuricemia, we evaluated the hypouricemic effects of ISO in vivo and in vitro, and explored the underlying mechanisms. Yeast extract-induced hyperuricemia animal model as well as hypoxanthine and xanthine oxidase (XOD) co-induced high uric acid L-O2 cell model and enzymatic experiments in vitro were selected. The XOD activity and uric acid (UA) level were inhibited after the treatment of ISO in vitro and in vivo. Furthermore, serum creatinine (CRE) and blood urea nitrogen (BUN) levels were also significantly reduced and liver damage was recovered in pathological histology after the ISO administration in hyperuricemia animal model. The results of mechanism illustrated that protein expressions such as XOD, toll-like receptor 4 (TLR4), cathepsin B (CTSB), NLRP3, and its downstream caspase-1 as well as interleukin-18 (IL-18) were markedly downregulated by ISO intervention in vitro and in vivo. Our results suggest that ISO exerts a urate-lowering effect through inhibiting XOD activity and regulating TLR4-NLRP3 inflammasome signal pathway, thus representing a promising candidate therapeutic agent for hyperuricemia. Both animal models and in vitro experiments suggested that ISO may effectively lower uric acid produce. The mechanism might be the inhibition of XOD activity and NLRP3 inflammasome of upregulation.

Therapeutic Duration and Extent Affect the Effect of Moxibustion on Depression-Like Behaviour in Rats via Regulating the Brain Tryptophan Transport and Metabolism.

Moxibustion has been widely accepted as an alternative therapy for major depressive disease (MDD). However, the efficacy of moxibustion treatment on MDD is highly variable because of its irregular operation. This study was designed to investigate how therapeutic duration and extent influence the anti-depression effect of moxibustion and the underlying mechanism involved. Rats with lipopolysaccharide-induced depression-like behavior were treated by moxibustion treatment. The anti-depression effect was determined by forced swimming test and open field test. Tryptophan (Trp) transport and its metabolism to serotonin (5-HT) and kynurenine (Kyn) were evaluated to explore the anti-depression mechanism. The results showed that moxibustion treatment could alleviate the depression-like behavior in rats. Trp transport and 5-HT generation were significantly increased, and the Trp-Kyn pathway was moderately inhibited by moxibustion. Prolonged therapy could be beneficial to the anti-depression effect by promoting the brain uptake of Trp and shifting the Trp metabolism to 5-HT. An enhanced therapeutic extent could increase 5-HT generation. In conclusion, this study determined that the anti-depression effect of moxibustion involves improved Trp transport and metabolism. The therapeutic duration benefits antidepressant effects, but the complex influence of the therapeutic extent on moxibustion efficacy requires further studies.

Combinatorial effect of blueberry extracts and oxaliplatin in human colon cancer cells.

The use of natural compounds to potentiate the effect of drugs and lower their adverse effects is an active area of research. The objective is to determine the effect of combined blueberry extracts (BE) and oxaliplatin (OX) in colon cancer cells. The results demonstrated that treatments of BE/OX showed inhibitory effects on HCT-116 cell and nontoxic effect on CCD-18Co normal colon cells. Flow cytometry analysis indicated that treatment with the BE, OX or in combination could induce G0/G1 cell cycle arrest, apoptosis, increase of reactive oxygen species, and induce loss of mitochondrial membrane potential in HCT-116 cells. Furthermore, after treatments, the expression of inflammatory cytokines was decreased, cyclin D1 and CDK4 were decreased; caspases-3 and 9 were activated; the Akt/Bad/Bcl-2 pathway was modulated. Moreover, the combination treatment had a considerably higher growth inhibitory effect on human colon cancer HCT-116 cells than that of BE or oxaliplatin alone. Our results showed that BE increased the anticolon cancer effect of OX making it an attractive strategy as adjuvant therapy to potentially reduce the adverse side effects associated with chemotherapeutic drugs.

Sulfur cycling in freshwater sediments: A cryptic driving force of iron deposition and phosphorus mobilization.

Sulfur cycling in freshwater ecosystems has been previously considered minor, and the direct evidence of its impacts on iron and phosphorus cycles in freshwater sediments remains unclear. In this study, mesocosms with amended acetate and various sulfate concentrations (1.5-3.0 mmol L-1) were set up to investigate sulfur cycling and its influences on iron-rich freshwater sediments. Acetate addition induced hypoxia and provided substrates, which stimulated the sulfur cycling with evidence of SO42- decline, ΣS2-, S0 increase and corresponding variations of sulfate-reducing bacteria (SRB) and sulfur-oxidizing bacteria. Meanwhile, the growth of iron-reducing bacteria (IRB) was suppressed, and lower Fe(II) release was correspondingly related to larger SRB abundance at higher sulfate level, indicating that microbial iron reduction might be blocked by SRB activities. However, continuous dissolution of Fe(III) oxides and generation of iron sulfides were observed, suggesting that sulfide-mediated chemical iron reduction (SCIR) became the dominant iron-reducing pathway, and Fe(II) was buried as iron sulfides instead of released to water column, which resulted in a transition of iron cycling into unidirectional SCIR. Consequently, continuous dissolution of Fe(III) oxides led to significant increase of PO43- concentration in the water column and sediment pore-water, revealing the phosphorus mobility in sediments derived from the SCIR process. To note, sustained accumulation of iron sulfides was observed even without ΣS2- presence, suggesting that ΣS2- precipitation occurred prior to diffusion. Thus, ΣS2--missing sulfur cycling seemed "cryptic" in this study. To highlight, the transition of the iron-reducing pathway and resulting PO43- release can be induced even under current sulfate level of Lake Taihu, and elevated sulfate levels could significantly intensify SCIR and phosphorus mineralization. Thus, the stimulated iron deposition and the resulting phosphorus release derived from the sulfur cycling should be paid more attention to in the treatment of eutrophic freshwater ecosystems.

Combined treatment with Dendrobium candidum and black tea extract promotes osteoprotective activity in ovariectomized estrogen deficient rats and osteoclast formation.

AIMS: Dendrobium candidum (DC) and black tea, are traditional chinese drinks, which contain multiple active ingredients. However, whether or not the combination of these two ingredients can improve osteoporosis remains unknown. This study therefore aimed to examine the effects of the combination of DC and black tea extract (BTE) on osteoporosis. MAIN METHODS: Ovariectomy (OVX)-induced osteoporosis in vivo as well as receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis in vitro was selected. KEY FINDINGS: Results showed that OVX rats that were treated orally with a DC and BTE combination for 12 weeks maintained their calcium (Ca) and phosphorus (P) homeostasis and exhibited significantly enhanced estradiol (E2) and OPG levels. This combination treatment also simultaneously reduced levels of interleukin (IL)-1ß, IL-6 and improved the organ coefficients of the uterus and femur as well as BMD and BMC in OVX rats. In addition, this DC and BTE combination suppressed osteoclast differentiation in the RANKL-stimulated osteoclastogenesis of RAW 264.7 cells and effectively inhibited the expression of osteoclast-associated genes and proteins. The results of this study further highlight the fact that a combination of DC and BTE improved ovariectomy-induced osteoporosis in rats and suppressed RANKL-stimulated osteoclastogenesis in RAW 264.7 cells. SIGNIFICANCE: This combination also significantly alleviated osteoporosis when compared to the alternative sole treatments above, due to synergistic effects among components. One partial mechanism of this combination might be the inhibition of osteoclast proliferation and the regulation of NFATC1/c-Fos expression.

Climate warming and cyanobacteria blooms: Looks at their relationships from a new perspective.

Climate warming and eutrophication are regarded as two important contributors to the occurrence of cyanobacteria blooms in aquatic ecosystems. However, the feedback of cyanobacteria blooms to climate warming and eutrophication is not fully clear. In this study, a microcosm system was established to simulate the decomposition processes of cyanobacteria blooms. It was observed that a large amount of nitrogen and phosphorus was released into the overlying water, and the concentrations of nitrogen and phosphorus were increased with the amount of added cyanobacteria bloom biomass addition. Subsequently, these released nutrients became available for primary production and intensified the eutrophic state of freshwater lakes. During the decomposition of cyanobacteria blooms, the microenvironment acquired low DO, low pH, and reductive conditions. Together with abundant organic matter in the water column and sediment, a large amount of CH4 and CO2 produced through organic matter mineralization, in which CH4 was the dominant fraction, occupied 50%-92% in mass of emitted carbon. Furthermore, a certain amount of N2O, probably underestimated, was produced with a strong greenhouse effect, even though its magnitude was small. These observations clarify that the feedbacks among cyanobacteria blooms formation and climate warming as well as the eutrophication of freshwater lakes are not unidirectional, but bidirectional. Given that climate warming enhanced the occurrence of cyanobacteria blooms, it was proposed that there are two vicious loops between cyanobacteria blooms, lake eutrophication and climate warming, which should be considered in the future management of aquatic ecosystems.

Physiological and proteomic responses of diploid and tetraploid black locust (Robinia pseudoacacia L.) subjected to salt stress.

Tetraploid black locust (Robinia pseudoacacia L.) is adaptable to salt stress. Here, we compared morphological, physiological, ultrastructural, and proteomic traits of leaves in tetraploid black locust and its diploid relatives under salt stress. The results showed that diploid (2×) plants suffered from greater negative effects than those of tetraploid (4×) plants. After salt treatment, plant growth was inhibited, photosynthesis was reduced, reactive oxygen species, malondialdehyde content, and relative electrolyte leakage increased, and defense-related enzyme activities decreased in 2× compared to those in 4×. In addition, salt stress resulted in distorted chloroplasts, swollen thylakoid membranes, accumulation of plastoglobules, and increased starch grains in 2× compared to those in 4×. However, 4× developed diverse responses under salt stress. A comparative proteomic analysis revealed that 41 and 37 proteins were differentially expressed in 2× and 4×, respectively. These proteins were mainly involved in photosynthesis, stress and defense, energy, metabolism, transcription/translation, and transportation. Distinct patterns of protein changes between 2× and 4× were analyzed. Collectively, our results suggest that the plants showed significantly different responses to salt stress based on ploidy level of the plant. The 4× possessed a better salt protection mechanism than that of 2×, suggesting salt tolerance in the polyploid plant.