Liuwei Dihuang pills attenuate ovariectomy-induced bone loss by alleviating bone marrow mesenchymal stem cell (BMSC) senescence via the Yes-associated protein (YAP)-autophagy axis.

CONTEXT: Liuwei Dihuang pill (LWDH) has been used to treat postmenopausal osteoporosis (PMOP). OBJECTIVE: To explore the effects and mechanisms of action of LWDH in PMOP. MATERIALS AND METHODS: Forty-eight female Sprague-Dawley rats were divided into four groups: sham-operated (SHAM), ovariectomized (OVX), LWDH high dose (LWDH-H, 1.6 g/kg/d) and LWDH low dose (LWDH-L, 0.8 g/kg/d); the doses were administered after ovariectomy via gavage for eight weeks. After eight weeks, the bone microarchitecture was evaluated. The effect of LWDH on the differentiation of bone marrow mesenchymal stem cells (BMSCs) was assessed via osteogenesis- and lipogenesis-induced BMSC differentiation. The senescence-related biological indices were also detected using senescence staining, cell cycle analysis, quantitative real-time polymerase chain reaction and western blotting. Finally, the expression levels of autophagy-related proteins and Yes-associated protein (YAP) were evaluated. RESULTS: LWDH-L and LWDH-H significantly modified OVX-induced bone loss. LWDH promoted osteogenesis and inhibited adipogenesis in OVX-BMSCs. Additionally, LWDH decreased the positive ratio of senescence OVX-BMSCs and improved cell viability, cell cycle, and the mRNA and protein levels of p53 and p21. LWDH upregulated the expression of autophagy-related proteins, LC3, Beclin1 and YAP, in OVX-BMSCs and downregulated the expression of p62. DISCUSSION AND CONCLUSIONS: LWDH improves osteoporosis by delaying the BMSC senescence through the YAP-autophagy axis.

To study the protective effect of Huangqi Baihe Granules on Radiation brain injury based on network pharmacology and experiment.

ETHNOPHARMACOLOGICAL RELEVANCE: Huangqi baihe Granules (HQBHG), which is a key Chinese medical prescription, has a remarkable efficacy in oxidative stress and inflammation. Nevertheless, the therapeutic effect on Radiation brain injury (RBI) has rarely been studied. AIM OF THE STUDY: The study aimed to verify the effect of HQBHG against RBI and explore its potential mechanism. METHODS: The potential targets and mechanisms of HQBHG against RBI were predicted by network pharmacology and verified by established rat model of RBI Firstly, the therapeutic effect of HQBHG in RBI was confirmed by water maze test, HE staining and Enzyme-linked immunosorbent assay (ELISA). Secondly, the potential critical anti-RBI pathway of HQBHG was further explored by water maze, HE staining, immunofluorescence assays, ELISA and western blot. RESULTS: A total of 43 HQBHG anti-RBI targets were obtained. Gene Ontology (Go) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional annotations showed that the treatment of HQBHG in RBI might be mainly related to oxidative stress, inflammation and PI3K/AKT pathway. Experimental studies have indicated that HQBHG can improve spatial learning and memory ability, alleviate pathological damage of brain tissue in RBI of rats. HQBHG also can down-regulate the levels of IL-1ß, TNF-α, ROS and MDA, meanwhile, GSH was significantly up-regulated. In addition, the HQBHG can increase the protein expression phosphorylations PI3K (p-PI3K), phosphorylations AKT(p-AKT) and Nrf2 in the brain tissue of RBI. CONCLUSION: HQBHG may alleviated RBI by regulated oxidative stress and inflammatory response through PI3K/AKT/Nrf2 pathway.

Deciphering the Active Compounds and Mechanisms of HSBDF for Treating ALI via Integrating Chemical Bioinformatics Analysis.

The Huashi Baidu Formula (HSBDF), a key Chinese medical drug, has a remarkable clinical efficacy in treating acute lung injury (ALI), and it has been officially approved by the National Medical Products Administration of China for drug clinical trials. Nevertheless, the regulated mechanisms of HSBDF and its active compounds in plasma against ALI were rarely studied. Based on these considerations, the key anti-inflammatory compounds of HSBDF were screened by molecular docking and binding free energy. The key compounds were further identified in plasma by LC/MS. Network pharmacology was employed to identify the potential regulatory mechanism of the key compounds in plasma. Next, the network pharmacological prediction was validated by a series of experimental assays, including CCK-8, EdU staining, test of TNF-α, IL-6, MDA, and T-SOD, and flow cytometry, to identify active compounds. Molecular dynamic simulation and binding interaction patterns were used to evaluate the stability and affinity between active compounds and target. Finally, the active compounds were subjected to predict pharmacokinetic properties. Molecular docking revealed that HSBDF had potential effects of inhibiting inflammation by acting on IL-6R and TNF-α. Piceatannol, emodin, aloe-emodin, rhein, physcion, luteolin, and quercetin were key compounds that may ameliorate ALI, and among which, there were five compounds (emodin, aloe-emodin, rhein, luteolin, and quercetin) in plasma. Network pharmacology results suggested that five key compounds in plasma likely inhibited ALI by regulating inflammation and oxidative damage. Test performed in vitro suggested that HSBDF (0.03125 mg/ml), quercetin (1.5625 µM), emodin (3.125 µM), and rhein (1.5625 µM) have anti-inflammatory function against oxidative damage and decrease apoptosis in an inflammatory environment by LPS-stimulation. In addition, active compounds (quercetin, emodin, and rhein) had good development prospects, fine affinity, and stable conformations with the target protein. In summary, this study suggested that HSBDF and its key active components in plasma (quercetin, emodin, and rhein) can decrease levels of pro-inflammatory factors (IL-6 and TNF-α), decrease expression of MDA, increase expression of T-SOD, and decrease cell apoptosis in an inflammatory environment. These data suggest that HSBDF has significant effect on anti-inflammation and anti-oxidative stress and also can decrease cell apoptosis in treating ALI. These findings provided an important strategy for developing new agents and facilitated clinical use of HSBDF against ALI.

Three-stage treatment for nitrogen and phosphorus recovery from human urine: Hydrolysis, precipitation and vacuum stripping.

Source separation of human urine has not been widely adopted because of scaling on urine collecting fixtures and lack of verified technologies for on-site utilization of waterless urine. This study investigated the effects of flushing liquid, temperature and urease amendment on hydrolysis of urea to ammonia, explored ammonia recovery via vacuum stripping in connection with phosphorus recovery via struvite precipitation in different sequences, and performed economic analysis of a proposed nutrient recovery strategy. It was found that acetic acid could be dosed at 0.05-0.07 N to flush urine-diverting toilets and urinals for hygiene and prevention of scaling. However, a high dosage of 0.56 N completely inhibited urea hydrolysis. Source-separated urine could be stored at 25 °C with ample urease for complete urea hydrolysis within approximately 20 h. Fully hydrolyzed waterless urine contained 9.0-11.6 g/L ammonia-N, 0.53-0.95 g/L phosphate-P and only 2.3-9.1 mg/L magnesium. When magnesium was supplemented to attain the optimum Mg2+: PO43- molar concentration ratio of 1.0 in hydrolyzed urine, batch operation of a pilot-scale air-lift crystallizer removed 93-95% of phosphate and produced 3.65-4.93 g/L struvite in 1-5 h. Batch operation of a pilot-scale vacuum stripping - acid absorption system for 12 h stripped 72-77% of ammonia and produced 37.6-39.7 g/L (NH4)2SO4. Compared with the ammonia → phosphorus recovery sequence, the struvite precipitation → vacuum stripping sequence produced more struvite and ammonium sulfate. The strategy of urea hydrolysis → struvite precipitation → vacuum stripping of ammonia is a sustainable alternative to the conventional phosphorus fertilizer production and ammonia synthesis processes.

NDUFAB1 confers cardio-protection by enhancing mitochondrial bioenergetics through coordination of respiratory complex and supercomplex assembly.

The impairment of mitochondrial bioenergetics, often coupled with exaggerated reactive oxygen species (ROS) production, is a fundamental disease mechanism in organs with a high demand for energy, including the heart. Building a more robust and safer cellular powerhouse holds the promise for protecting these organs in stressful conditions. Here, we demonstrate that NADH:ubiquinone oxidoreductase subunit AB1 (NDUFAB1), also known as mitochondrial acyl carrier protein, acts as a powerful cardio-protector by conferring greater capacity and efficiency of mitochondrial energy metabolism. In particular, NDUFAB1 not only serves as a complex I subunit, but also coordinates the assembly of respiratory complexes I, II, and III, and supercomplexes, through regulating iron-sulfur biosynthesis and complex I subunit stability. Cardiac-specific deletion of Ndufab1 in mice caused defective bioenergetics and elevated ROS levels, leading to progressive dilated cardiomyopathy and eventual heart failure and sudden death. Overexpression of Ndufab1 effectively enhanced mitochondrial bioenergetics while limiting ROS production and protected the heart against ischemia-reperfusion injury. Together, our findings identify that NDUFAB1 is a crucial regulator of mitochondrial energy and ROS metabolism through coordinating the assembly of respiratory complexes and supercomplexes, and thus provide a potential therapeutic target for the prevention and treatment of heart failure.

Highly Sensitive and Selective Determination of Tertiary Butylhydroquinone in Edible Oils by Competitive Reaction Induced "On-Off-On" Fluorescent Switch.

As one of most common synthetic phenolic antioxidants, tertiary butylhydroquinone (TBHQ) has received increasing attention due to the potential risk for liver damage and carcinogenesis. Herein, a simple and rapid fluorescent switchable methodology was developed for highly selective and sensitive determination of TBHQ by utilizing the competitive interaction between the photoinduced electron transfer (PET) effect of carbon dots (CDs)/Fe(III) ions and the complexation reaction of TBHQ/Fe(III) ions. This novel fluorescent switchable sensing platform allows determining TBHQ in a wider range from 0.5 to 80 µg mL(-1) with a low detection limit of 0.01 µg mL(-1). Furthermore, high specificity and good accuracy with recoveries ranging from 94.29 to 105.82% in spiked edible oil samples are obtained with the present method, confirming its applicability for the trace detection of TBHQ in a complex food matrix. Thus, the present method provides a novel and effective fluorescent approach for rapid and specific screening of TBHQ in common products, which is beneficial for monitoring and reducing the risk of TBHQ overuse during food storage.

[Mechanism of Tongsaimai tablet for atherosclerosis based on network pharmacology].

Network pharmacology method was adopted in this study to explore the active compounds and mechanism of Tongsaimai tablets for atherosclerosis. In molecular docking and molecular-target protein network analysis, 97 molecules in Tongsaimai tablets showed good interaction with the atherosclerosis-related target protein (docking score ≥ 7), and 37 molecules of them could act on more than 2 targets (≥ 2) with higher betweenness, suggesting that these 37 molecules might be the main active compounds group in Tongsaimai tablets for atherosclerosis treatment. Furthermore, the predicted active compounds contained more flavonoids and saponins, reminding more attention should be paid on flavonoids and saponins in study of effective compounds and quality standards of Tongsaimai tablets. Targets network analysis showed that, the active compounds of Tongsaimai tablets could regulate inflammation, stabilize plaque, protect vascular endothelial cell, regulate blood lipid and inhibit blood coagulation through acting on the main 22 target proteins, such as Toll-like receptors (TLR1, TLR2), matrix metalloproteinase (MMP1, MMP2, MMP3, MMP9), angiotensin converting enzyme (ACE), leukotriene A4 hydrolase (LTA4-H), 5-lipoxidase (5-LOX), peroxisome proliferators-activated receptors (PPARα, PPARγ). These active compounds can participate in regulating different pathologic stages of atherosclerosis and thus treat atherosclerosis finally. This study revealed the main active compounds and possible mechanism of Tongsaimai tablets for treatment of atherosclerosis and meanwhile, verified the characteristics of multi-components, multi-targets and integral regulation for Tongsaimai tablets, providing theoretical references for the following systematic laboratory experiments on effective compounds and action mechanism of Tongsaimai Tablet.

[Study on anti-inflammation and immunoloregulation effect of Guizhi Fuling capsule ingredients using high content screening].

The present study sought to investigate the anti-inflammation and immunoloregulation effect of 17 Guizhi Fuling capsule ingredients. The anti-inflammatory ingredients on LPS-induced RAW264. 7 cell injury were assessed with ELISA and immunofluorescence. The release of IL-1ß, TNF-α, PGE2 were detected with ELISA and the expression of COX-2 was detected with immunofluorescence. The effects of them on promoting splenic lymphocyte proliferation were assessed with MTT and Hoechst 33342 staining method. The results showed that 15 ingredients had obviously anti-inflammatory activity on LPS- induced injury and play the immunoloregulation roles. This study suggested that the 15 ingredients may be the active ingredients on pelvic infection.

Catecholaminergic-induced arrhythmias in failing cardiomyocytes associated with human HRCS96A variant overexpression.

The histidine-rich calcium binding protein (HRC) Ser96Ala polymorphism was shown to correlate with ventricular arrhythmias and sudden death only in dilated cardiomyopathy patients but not in healthy human carriers. In the present study, we assessed the molecular and cellular mechanisms underlying human arrhythmias by adenoviral expression of the human wild-type (HRC(WT)) or mutant HRC (HRC(S96A)) in adult rat ventricular cardiomyocytes. Total HRC protein was increased by ∼50% in both HRC(WT)- and HRC(S96A)-infected cells. The HRC(S96A) mutant exacerbated the inhibitory effects of HRC(WT) on the amplitude of Ca(2+) transients, prolongation of Ca(2+) decay time, and caffeine-induced sarcoplasmic reticulum Ca(2+) release. Consistent with these findings, HRC(S96A) reduced maximal sarcoplasmic reticulum calcium uptake rate to a higher extent than HRC(WT). Furthermore, the frequency of spontaneous Ca(2+) sparks, which was reduced by HRC(WT), was increased by mutant HRC(S96A) under resting conditions although there were no spontaneous Ca(2+) waves under stress conditions. However, expression of the HRC(S96A) genetic variant in cardiomyocytes from a rat model of postmyocardial infarction heart failure induced dramatic disturbances of rhythmic Ca(2+) transients. These findings indicate that the HRC Ser96Ala variant increases the propensity of arrhythmogenic Ca(2+) waves in the stressed failing heart, suggesting a link between this genetic variant and life-threatening ventricular arrhythmias in human carriers.

[Effects of soil factors on active component content of Chrysanthemum morifolium].

OBJECTIVE: To study the effects of soil factors on the active component content of Chrysanthemum morifolium and screen out the leading factors. METHOD: The active component of water soluble extracts, flavonoids, phenolic compounds and mineral elements were determined and chemical properties and mineral elements of soil were analyzed for studying the effects on Ch. morifolium through correlation, stepwise regression, path and grey correlation analysis. RESULT: Soil available P and K were the most important factors that affected the active component content of Ch. morifolium, followed by urease, phosphatase and invertase activities and organic matter. The mineral elements in Ch. morifolium and in soil correlated well, P and K were enriched in the plant mostly, followed by Cd, Ca, Zn, Cu. The main leading factors of mineral elements in soil were P and K, followed by Fe, Cu and Zn. CONCLUSION: Soil was one of the important factors which affected the active component content of Ch. morifolium.

[Effects of light intensity on physiological and biochemical characteristics of Chrysanthemum morifolium at vegetative stage].

OBJECTIVE: To study the effect of light intensity on physiological and biochemical characteristics of Chrysanthemum morifolium at the vegetative stage. METHOD: The dynamic response of physiological and biochemical indexes of Ch. morifolium were measured under different treatments (100%, 80%, 60%, 40% and 20% of the full sunlight) at the vegetative stage. RESULT: The physiological and biochemical indexes of Ch. morifolium showed dynamic changes with the progress of growth and the increase of the treatment time. The soluble sugar content decreased when the light intensity reduced, and had a significant positive correlation with the light intensity. Soluble protein content rose firstly and fell later, malondialdehyde content increased, superoxide dismutase and catalase activity decreased initially and increased afterwards. CONCLUSION: Proper shading benefits the nitrogen accumulation of Ch. morifolium at the vegetative stage, and reduces the strength of stress condition. The suitable light environment for growth of Ch. morifolium at the vegetative stage is about 80%-60% of full sunlight and the optimum treatment time is 20-40 days.

[Determination of quercetin, luteolin, apigenin and acacetin in Flos Chrysanthemi Indici by RP-HPLC].

OBJECTIVE: To develop a RP-HPLC method for the determination of quercetin, luteolin, apigenin and acacetin in Flos Chrysanthemi indici. METHOD: An Eclipse XDB-C18 column (4.6 mm x 250 mm, 5 microm) was used at 25 degrees C with the mobile phases of methanol-0.2% phosphatic acid in a gradient manner. The flow rate was set at 1.0 mL x min(-1). The detection wavelength was 350 nm. RESULT: The linear response ranged from 1.02-20.48 mg x L(-1) for quercetin (r = 0.9994, n = 5), 1.03-20.54 mg x L(-1) for luteolin (r = 0.9992, n = 5), 1.12-22.40 mg x L(-1) for apigenin (r = 0.9995, n = 5), 1.01-20.22 mg x L(-1) for acacetin (r = 0.9998, n = 5), respectively. Recoveries were 101.3% with RSD 1.3% for quercetin, 100.62% with RSD 1.4% for luteolin, 98.42% with RSD 1.7% for apigenin and 99.02% with RSD 0.8% for acacetin. A significant difference (alpha = 0.01) among the contents of four flavonoids and total flavonoids was found. CONCLUSION: The method is quick, simple and repeatable for simultaneous determination of quercetin, luteolin, apigenin and acacetin in Flos Chrysanthemi Indici.

[Effects of light intensity on growth and photosynthetic characteristics of Chrysanthemum morifolium].

OBJECTIVE: To investigate the effect of light intensity on growth and photosynthetic of Chrysanthemum morifolium. METHOD: The growth characteristics of C. morifolium were measured under different treatments (100%, 80%, 60%, 40% and 20% of full sunlight). The photosynthetic characteristics and chlorophyll fluorescence parameters of leaves under different light intensity were determined by a LI-6400 photosynthesis system and a PAM-2100 chlorophyll fluorescence system. RESULT: With the reduction of irradiance, the diameter of the stem reduced, plant height, leaf length, leaf width and length/width raised, assimilation product increased; Content of photosynthetic pigment increased between light intensity 100%-40% reduced under 20% treatment, chlorophyll a/b decreased. Light compensation point (LCP), apparent quantum yield (AQY) increased first and reduced later, photosynthesis rate (P(n)), stomatal conductance (G(s)), intercellular CO2 concentration (C(i)) and transpiration rate (T(r)) decreased, stomatal limitation value (L(s)) rose. Chlorophyll fluorescence parameters F(v)/F(m) increased, phiPS II, F(v)'/F(m)', ETR and qP increased between irradiance 100%-60%, NPQ decreased, such as phiPS II decreased and NPQ increased when irradiance was lower than 40%. CONCLUSION: Weak light condition was unfavorable to the growth of C. morifolium and the light conditions of culture should be control between 80%-60% of full sunlight.

[Study on seed quality test and quality standard of Prunella vulgaris].

OBJECTIVE: To study seed quality test, and establish seed quality classification standard of Prunella vulgaris. METHOD: The seed quality and planting quality of P. vulgaris from different population were measured and these indexes were analyzed by K-clustering. RESULT AND CONCLUSION: The seed purity, 1000 seeds weight, seed moisture content, seed viability of the seed of P. vulgaris from different population were measured, and the seed quality classification standard of P. vulgaris.

[Effects of seed priming on vigor of Prunella vulgaris seeds].

OBJECTIVE: To select an effective way to enhance vigor of Prunella vulgaris seeds. METHOD: Three population seeds were treated at the 20 degrees C and dark enviroment. RESULT: Priming with 20% - 30% PEG and 200 - 400 mg x L(-1) GA3 could enhance seeds germination and vigor. Germination percentage of three population seeds treated with 0. 6% - 3.0% NaCl reduced, but they started to germinate in advance. Treated with 0.6% - 2.4% KNO3-KH2PO4, germination rate and vigor of seeds in Zijinshan and Pan' an both increased and the one in Bozhou decreased. CONCLUSION: Vigor of P. vulgaris seed treated with PEG and GA3 under proper concentration increases, while treated with KNO3-KH2PO, and NaCl low vigor seeds germination rate reduces.

Ultrasonic treatment on activated sewage sludge from petro-plant for reduction.

In this essay, the influences of low frequency ultrasound (20 kHz) on dewater ability and anaerobic digestion behaviors of activated sewage sludge, obtained from Yangzi Water Treatment Plant, Yangzi Petrochemical Corporation, were discussed. Ultrasound pretreatment could enhance the filtration progress and decrease the moisture content of the sludge from 99% to 80%. Together with flocculant, the ultrasound pretreatment decreased the specific filtration resistance (SFR) of the sludge from 3.59 x 10(12) m/kg to 1.18 x 10(12) m/kg and saved about 25-50% of the flocculant dosage. Bound water of the sludge was measured by dilatometer. After 2-4 min treatment of ultrasound under intensity of 400 W/m(2), the bound water of sludge decreased from 16.7 g/g (dry base) to above 2.0 g/g (dry base). Ultrasound pretreatment could also enhance digestion and reduce digestion time. To the same resolution ratio, such as 49%, the digestion time of sludge with ultrasound pretreatment was 7 days less than that without ultrasound. Proper ultrasound pretreatment could also improve the dewater ability of the digested sludge, the final moisture of which was 73.7%.

Modulation of secretory functions in epithelia by adenovirus capsid proteins.

To evaluate the safety of adenovirus-derived capsid proteins for ocular gene delivery, we have investigated their effects on the morphology and function of the acinar epithelial cells of the lacrimal gland. These cells are responsible for basal and stimulated release of proteins and electrolytes into ocular fluid, a process essential in maintaining the health of the ocular surface. Acinar epithelial cells from rabbit lacrimal gland were exposed to one of two adenovirus serotype 5 capsid proteins, penton or knob (the carboxy-terminal fragment of the fiber capsid protein). Sustained (16-18 h) exposure to the penton at 20 microg/ml was associated with major changes in the organization of the regulated secretory pathway and cytoskeleton. These changes included an apparent loss of mature secretory vesicles enriched in rab3D around the apical lumen as well as a depletion of apical actin. The microtubule array in penton-treated acini also exhibited bundling and disorganization. None of these effects were elicited by exposure to knob protein. Penton treatment also caused a significant (p < or = 0.05) increase and decrease in basal and carbachol-stimulated release, respectively, of bulk protein. Competition studies showed that RGD peptide partially prevented the penton-induced changes in rab3D-enriched secretory vesicles and actin filaments. These findings suggest that the adenovirus penton protein compromises normal acinar secretory compartment organization and function and that these changes are due at least partly to penton-integrin interactions.