Biochar addition regulates soil phosphorus fractions and improves release of available phosphorus under freezing-thawing cycles.

Currently, the shortage of phosphorus resources is becoming more and more serious. In general, phosphorus fertilizer is poorly utilized in soil and tends to gradually accumulate. Freezing-thawing cycles (FT) are seasonal phenomenon occurring in high latitudes and altitudes regions, which have obvious influence on the form of phosphorus in soil. This study investigates the effect of biochar on soil physicochemical properties, phosphorus form and availability under FT and thermostatic incubation (TH) condition. Compared with treatment without biochar, 4 % biochar addition increased the soil pH value, electrical conductivity, organic matter and Olsen-P of soil by a maximum of 0.76, 285.55 µS/cm, 28.60 g/kg and 139.27 mg/kg, respectively. Moreover, according to Hedley-P classification results, under FT condition, the content of labile phosphorus pool is always higher than those under TH. FT may promote the conversion of phosphorus from other fractions to labile phosphorus pool. Redundancy analysis results show that biochar addition and FT can not only directly change the soil phosphorus pool, but also alter the soil physicochemical properties and microbial community, which further affect the adsorption and mineralization of phosphorus in soil. The results of this study will be devoted to understanding the changes in soil phosphorus fractions under the effects of biochar addition and FT, providing references for agricultural production in areas where FT occur.

Metal-based adsorbents for water eutrophication remediation: A review of performances and mechanisms.

Controlling eutrophication requires satisfying stringent phosphorus concentration standards. Metal-based adsorbents can effectively remove excess phosphorus from water bodies and achieve ultra-low phosphorus concentration control for wastewater. This review focuses on the material properties and phosphorus removal mechanism of metal-based adsorbents (Fe, Al, Ca, Mg, La). There are significant differences in physical and chemical properties of different metal materials, due to the different preparation methods and synthetic materials. The main factors affecting phosphorus removal performance include particle size, crystal structure and pHPZC. Smaller particle size, more disordered crystal structure and higher pHPZC are more favorable for phosphorus removal. The main mechanism of phosphorus removal by metal-based adsorbents is ligand exchange, which makes it exhibit excellent adsorption capacity, fast kinetics and well selectivity for phosphate. In addition, in order to improve the phosphorus removal performance, the surface properties of the adsorbent (e.g., surface charge, surface area, and functional groups) can be effectively improved by dispersion of biochar carriers or combination of multiple metal materials. In further studies, we should improve the absorption capacity of the adsorbent under high pH conditions and the resistance to coexisting ion interference. Finally, in order to ensure the effective application of metal-based adsorbents in the phosphorus removal field, experimental scale should be expanded in future work to suit the actual water body conditions.

Effects of biochar on transport and retention of phosphorus in porous media: Laboratory test and modeling.

Given the complexity of soil components, a detailed understanding of the effects of single factors on phosphorus transport and retention will play a key role in understanding the environmental effects of phosphorus. In this work, quartz sand columns (considering five factors: doping rate, pH, particle size, ionic strength and cation type), combined with a two-site nonequilibrium transport model (TSM), were used to investigate phosphate (P) transport behavior. The results show that changes in doping ratio (0.4%-1.6%) and pH (5-9) have a notable effect on the transport of P, while, particle size of quartz sand hardly impacts the transport. When biochar was added at 1.6%, the surface of biochar increased the P fixation rate by about 37% through direct interaction with phosphate and bridging action with metal ions. As the morphology of P changed under different pH conditions, a part of P was immobilized in the form of precipitation. The immobilization of P was further enhanced with the increase of ionic strength. Compared with the direct interaction of P with biochar in Na+ solution, Ca2+ and Mg2+ solutions are more likely to adsorb P. Meanwhile, the TSM model also fits the transport behavior well. This study provides a perspective for evaluating the environmental behavior of P in the porous media interaction with biochar.

[Scr eening of key genes and pathways of brain aging and prediction of potential effective Chinese medicinals: based on bioinformatics].

Microarray data of hippocampal tissue(HC) of the cognitively intact elderly(60-99 years old) were compared with those of the middle-aged and the young(20-59 years old) by bioinformatics techniques to initially screen out differentially expressed genes(DEGs) and then predict potential effective Chinese medicinals for the treatment of brain aging. The gene expression profile(accession: GSE11882) was downloaded from the Gene Expression Omnibus(GEO) and DEGs were screened based on R package. The key DEGs were identified by STRING, Cytoscape and the plug-in, followed by Gene Ontology(GO) term and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analysis. Then the key genes and the medical ontology information retrieval platform(Coremine Medical) were mapped against each other to single out the Chinese medicinals for the treatment of brain aging and construct the " Chinese medicinal-active constituent-target" network. Among the resultant 268 DEGs(246 down-regulated and 22 up-regulated), the 15 key genes were mainly involved in biological processes such as leukocyte migration, neutrophil activation, cell chemotaxis, microglia activation and response to external stimulus, and pathways such as inflammatory process, immune response, cytokine-cytokine receptor interaction, PI3 K-Akt signaling pathway, Rap1 signaling pathway, chemokine signaling pathway and Toll-like receptor signaling pathway. The potential effective Chinese medicinals were Notoginseng Radix et Rhizoma, Ginseng Radix et Rhizoma, Salviae Miltiorrhizae Radix et Rhizoma and Astragali Radix. The analysis of DEGs and key genes enhances the understanding of the mechanisms of brain aging. This study provides potential gene targets and ideas for the development of Chinese medicine for brain aging.

[Mechanism of extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma on SIRT1 autophagy pathway of endothelial cell senescence induced by hydrogen peroxide].

This study aims to explore the effect of extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma, and Chuanxiong Rhizoma(hereinafter referred to as GNS) on the SIRT1-autophagy pathway of endothelial cell senescence induced by hydrogen peroxide(H_2O_2). To be specific, vascular endothelial cells were classified into the blank control group(control), model group(model), model + DMSO group(DMSO), resveratrol group(RESV), and GNS low-dose(GNS-L), medium-dose(GNS-M), and high-dose(GNS-H) groups. They were treated with H_2O_2 for senescence induction except the control. After intervention of cells in each group with corresponding drugs for 24 h, cell growth status was observed under an inverted microscope, and the formation of autophagosome under the transmission electron microscope. In addition, the changes of microtubule-associated protein 1 light chain 3ß(LC3 B) were detected by immunofluorescence staining. The autophagy flux was tracked with the autophagy double-labeled adenovirus(mRFP-GFP-LC3) fusion protein. Dansylcadaverine(MDC) staining was employed to determine the autophagic vesicles, and Western blot the expression of sirtuin 1(SIRT1), ubiquitin-binding protein p62, and LC3Ⅱ. After H_2O_2 induction, cells demonstrated slow growth, decreased adhesion ability, raised number of SA-ß-gal-stained blue ones, a certain number of autophagosomes with bilayer membrane and secondary lysosomes in the cytoplasm, and slight rise of autophagy flux level. Compared with the model group, GNS groups showed improved morphology, moderate adhesion ability, complete and smooth membrane, decreased SA-ß-gal-stained blue cells, many autophagosomes, autophagic vesicles, and secondary lysosomes in the cytoplasm, increased autophagolysosomes, autophagy flux level, and fluorescence intensity of LC3 B and MDC, up-regulated expression of SIRT1 and LC3Ⅱ, and down-regulated expression of p62, suggesting the improvement of autophagy level. GNS can delay the senescence of vascular endothelial cells. After the intervention, the autophagy flux and related proteins SIRT1, LC3Ⅱand p62 changed significantly, and the autophagy level increased significantly. However, EX527 weakened the effect of Chinese medicine in delaying vascular senescence. GNS may delay the senescence of vascular endothelial cells through the SIRT1 autophagy pathway.

Sustainable advances on phosphorus utilization in soil via addition of biochar and humic substances.

The intervention of human in phosphorus pool seems to be a vicious circle. The rapid population growth leads to the global food shortage, which leads to the massive use of phosphate fertilizer and the continuous exploitation of phosphate rocks. With the massive loss and fixation of phosphate fertilizer in the soil, the unavailable phosphorus in the soil becomes superfluous, while the phosphate mineral resources turn to scarce. Interestingly, exogenous carbonaceous materials, notably, biochar and humic substances, have been widely used as soil conditioners in agricultural production up to date, among other actions to interfere with the balance between the different phosphate species, which offer effective roles for increasing soil available phosphorus. This article reviews the regulation mechanisms of biochar and humic substances on phosphorus availability and circulation, including improving soil physicochemical characteristics, regulating microbial community structure, and directly interacting with phosphorus to affect the fate of phosphorus in soil. Finally, the prospects for future research directions are made, and it is hoped that the review of this article can arouse people's attention to the current plight of agricultural production and provide some methods for improving the efficiency of phosphate fertilizer use in the future.

Mechanism of extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma on SIRT1 autophagy pathway of endothelial cell senescence induced by hydrogen peroxide / 中国中药杂志

This study aims to explore the effect of extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma, and Chuanxiong Rhizoma(hereinafter referred to as GNS) on the SIRT1-autophagy pathway of endothelial cell senescence induced by hydrogen peroxide(H_2O_2). To be specific, vascular endothelial cells were classified into the blank control group(control), model group(model), model + DMSO group(DMSO), resveratrol group(RESV), and GNS low-dose(GNS-L), medium-dose(GNS-M), and high-dose(GNS-H) groups. They were treated with H_2O_2 for senescence induction except the control. After intervention of cells in each group with corresponding drugs for 24 h, cell growth status was observed under an inverted microscope, and the formation of autophagosome under the transmission electron microscope. In addition, the changes of microtubule-associated protein 1 light chain 3β(LC3 B) were detected by immunofluorescence staining. The autophagy flux was tracked with the autophagy double-labeled adenovirus(mRFP-GFP-LC3) fusion protein. Dansylcadaverine(MDC) staining was employed to determine the autophagic vesicles, and Western blot the expression of sirtuin 1(SIRT1), ubiquitin-binding protein p62, and LC3Ⅱ. After H_2O_2 induction, cells demonstrated slow growth, decreased adhesion ability, raised number of SA-β-gal-stained blue ones, a certain number of autophagosomes with bilayer membrane and secondary lysosomes in the cytoplasm, and slight rise of autophagy flux level. Compared with the model group, GNS groups showed improved morphology, moderate adhesion ability, complete and smooth membrane, decreased SA-β-gal-stained blue cells, many autophagosomes, autophagic vesicles, and secondary lysosomes in the cytoplasm, increased autophagolysosomes, autophagy flux level, and fluorescence intensity of LC3 B and MDC, up-regulated expression of SIRT1 and LC3Ⅱ, and down-regulated expression of p62, suggesting the improvement of autophagy level. GNS can delay the senescence of vascular endothelial cells. After the intervention, the autophagy flux and related proteins SIRT1, LC3Ⅱand p62 changed significantly, and the autophagy level increased significantly. However, EX527 weakened the effect of Chinese medicine in delaying vascular senescence. GNS may delay the senescence of vascular endothelial cells through the SIRT1 autophagy pathway.

Effect of Ginseng Radix et Rhizoma， Notoginseng Radix et Rhizoma， and Chuanxiong Rhizoma Extract on Mitochondrial Oxidative Stress in Hydrogen Peroxide-induced Endothelial Cell Aging / 中国实验方剂学杂志

Objective:To observe the effect of Ginseng Radix et Rhizoma， Notoginseng Radix et Rhizoma， and Chuanxiong Rhizoma extract （GNC） on mitochondrial oxidative stress in hydrogen peroxide （H₂O₂）-induced aging of human umbilical vein endothelial cells （HUVECs）， and explore the therapeutic mechanism of GNC on aging HUVECs. Method:The HUVECs were classified into the control group （control）， H₂O₂ model group （H₂O₂）， H₂O₂ + DMSO group （DMSO， 1 mL·L^-1）， resveratrol group （Resv， 8 μmol·L^-1）， and low- （200 mg·L^-1）， medium- （300 mg·L^-1）， and high-dose （400 mg·L^-1） GNC （GNC-L， GNC-M， and GNC-H） groups. Except control group and H₂O₂ group， the other groups were intervened with corresponding agents. Subsequently， 300 μmol·L^-1 H₂O₂ was given to other groups except the control group for 4 h to induce aging， and then the cells were cultured in normal media for 24 h. The aging degree， cell cycle， and mitochondrial reactive oxygen species （mtROS） level were determined by SA-<italic>β</italic>-galactosidase （SA-<italic>β</italic>-Gal） staining， flow cytometry， and MitoSox red fluorescence staining， respectively. JC-10 was used as a fluorescent probe to detect the changes in mitochondrial membrane potential， and Western blot was performed to detect the expression of manganese superoxide dismutase （MnSOD） and p-p66 proteins. Result:The SA-<italic>β</italic>-gal staining results showed that H₂O₂ group had increased blue-stained cells compared with other groups （<italic>P</italic><0.01）. Compared with those in the control group， the ratio of G₀/G₁ phase cells significantly increased （<italic>P</italic><0.05） and that of G₂/M phase cells decreased （<italic>P</italic><0.05） in the H₂O₂ group. Compared with those in the H₂O₂ group， the proportion of G₀/G₁ cells decreased （<italic>P</italic><0.05） while that of G₂/M cells increased （<italic>P</italic><0.05） in GNC-H groups and Resv group. The fluorescence staining for determining mitochondrial ROS level showed that the H₂O₂ group had weakened fluorescence intensity than the control， GNC-H， and GNC-M groups （<italic>P</italic><0.05）. The mitochondrial membrane potential fluorescence intensity of the H₂O₂ group was weaker than that of the control， GNC-H， GNC-M， and GNC-L groups （<italic>P</italic><0.01）， as well as the Resv group （<italic>P</italic><0.05）. Western blot showed that the protein level of MnSOD was significantly lower in the H₂O₂ group than in the control， GNS-H， and GNS-M groups （<italic>P</italic><0.05）， whereas the protein level of p-p66 showed an opposite trend （<italic>P</italic><0.01）， indicating that the medication can alleviate the intracellular mitochondrial oxidative stress. Conclusion:GNC can delay the H₂O₂-induced aging of vascular endothelial cells. The GNC intervention significantly regulated the mitochondrial ROS， mitochondrial membrane potential， and related proteins MnSOD and p-p66 to alleviate oxidative stress. Chinese medicinal materials may delay the aging of vascular endothelial cells by inhibiting mitochondrial oxidative stress.

Ginseng-Sanqi-Chuanxiong (GSC) Extracts Ameliorate Diabetes-Induced Endothelial Cell Senescence through Regulating Mitophagy via the AMPK Pathway.

Vascular endothelial senescence induced by high glucose and palmitate (HG/PA) contributes to endothelial dysfunction, which leads to diabetic cardiovascular complications. Reduction of endothelial senescence may attenuate these pathogenic processes. This study is aimed at determining whether Ginseng-Sanqi-Chuanxiong (GSC) extracts, traditional Chinese medicine, can ameliorate human aortic endothelial cell (HAEC) senescence under HG/PA-stressed conditions and further explore the underlying mechanism. We found that GSC extracts significantly increased antisenescent activity by reducing the HG/PA-induced mitochondrial ROS (mtROS) levels in senescent HAECs. GSC extracts also induced cellular mitophagy formation, which mediated the effect of GSC extracts on mtROS reduction. Apart from this, the data showed that GSC extracts stimulated mitophagy via the AMPK pathway, and upon inhibition of AMPK by pharmacological and genetic inhibitors, GSC extract-mediated mitophagy was abolished which further led to reverse the antisenescence effect. Taken together, these data suggest that GSC extracts prevent HG/PA-induced endothelial senescence and mtROS production by mitophagy regulation via the AMPK pathway. Thus, the induction of mitophagy by GSC extracts may provide a novel therapeutic candidate for cardiovascular protection in metabolic syndrome.

Potential treatment methods targeting 2019-nCoV infection.

The novel coronavirus, 2019-nCoV, has quickly spread across the world and pose serious threat to public health because it can infect people very easily. The major clinical symptoms of 2019-nCoV infection include fever, dry cough, myalgia, fatigue, and diarrhea. The 2019-nCoV belongs to the betacoronavirus family, and gene sequencing results demonstrate that it is a single-stranded RNA virus, closely related to Severe Acute Respiratory Syndrome CoV (SARS-CoV) and Middle East Respiratory Syndrome CoV (MERS-CoV). It has been observed that the virus invades human body mainly through binding to angiotensin-converting enzyme 2 (ACE2) receptors similar to SARS-CoV and the main protease (Mpro) acts as a critical protease for digesting the polyprotein into functional polypeptides during the replication and transcription process of 2019-nCoV. In this review, we summarized the real-time information of 2019-nCoV treatment methods and mainly focused on the chemical drugs including lopinavir/ritonavir, chloroquine, hydroxychloroquine, arbidol, remdesivir, favipiravir and other potential innovative active molecules. Their potential targets, activity, clinical status and side effects are described. In addition, Traditional Chinese Medicine (TCM), Convalescent plasma therapy (CPT) and biological reagents available, as well as the promising vaccine candidates against 2019-nCoV are also discussed.

[Effect of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts on intestinal flora of vascular aging mice induced by high glucose and high lipid].

The aim of this paper was to observe the changes of intestinal flora in vascular aging mice, in order to explore the relationship between vascular aging and intestinal flora and the effects of extracts of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma on intestinal flora of vascular aging mice. A model of vascular aging in mice was induced through intrape-ritoneal injection with streptozotocin(STZ) combined with high-fat diet. Biochemical detection was performed on serum cholesterol(CHO), triglyceride(TG), high-density liptein cholesterol(HDL-C), low-density liptein cholesterol(LDL-C) and blood glucose(GLU). HE staining was used to detect mice thoracic aorta morphology, and the expressions of cyclin-dependent kinase inhibitor 2 A(p16) and cyclin-dependent kinase inhibitor 1 A(p21) protein in mice thoracic aorta were detected by Western blot. The 16 S rDNA gene of mice intestinal flora was detected by Illumina MiSeq high-throughput sequencing technology to explore the changes of intestinal flora in each group. The results demonstrated that the GLU level in low-dose and high-dose TCM groups decreased, but with unobvious changes in blood lipid indexes. Metformin could significantly decrease the levels of GLU(P<0.01), CHO and LDL-C in mice(P<0.05). Intravascular injury was not obvious in each drug group, and the expressions of p16 and p21 protein were significantly decreased(P<0.05). The intestinal flora of each group was mainly composed of Firmicutes(F) and Bacteroidetes(B) at the level of the phylum, but the B/F ratio was different from that of the youth group and the blank control group. The B/F ratio of the model group was significantly lower(P<0.01), and compared with the model group, the B/F ratio of the high-dose group and the metformin group was signi-ficantly higher(P<0.05). There were dominant and differential floras in the intestine of each group of mice. The results showed that extracts of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma could improve the intestinal flora structure and create a good intestinal environment by increasing the B/F ratio, which provides a new possible pathway for lowering blood glucose and blood lipids and delaying vascular aging.

Research Progress of Traditional Chinese Medicine for Yiqi Huoxue Huatan in Treatment of Atherosclerosis / 中国实验方剂学杂志

The application of clinical medication and basic research progress of traditional Chinese medicine （TCM） for Yiqi Huoxue Huatan in the treatment of atherosclerosis （AS） were summarized. According to the different pathogenic sites of AS， the clinical research progress of TCM for Yiqi Huoxue Huatan in the treatment of AS and the commonly used TCM for the treatment of AS were summarized. Astragali Radix， Salviae Miltiorrhizae Radix， Quinquefolium Panax， Cocos Wolf Poria， Atractylodis Rhizoma， Rosea Rhodiola， which were Yiqi herbs， were mostly used for the treatment of AS. Wallichii Ligusticum， Salviae Miltiorrhizae Radix， Notoginseng Radix， Paeoniae Rubra Radix， Paeoniae Alba Radix， Angelicae Sinensis Radix， Semen Persicae， Tinctorius Carthamus， Achyranthis Bidentatae Radix， tea root， which were Huoxue herbs， were mostly used for the treatment of AS. Huatan herbs， including Kirilowii Maxim Trichosanthes， Pinelliae Rhizama， Acorus Tatarinowii Schott， Citri Reticulatae Pericarpium， Cum Bile Arisaema， Silicea Bambusae Concretio， Aurantii Immaturus Fructus， Bamboo Juice， were commonly used for the treatment of AS. According to the findings， TCM for Yiqi Huoxue was mostly combined with insect medicine and rattan medicine for the treatment of carotid atherosclerosis， combined with TCM for promoting Qi， relieving pain， dissipating blood stasis and reducing phlegm for the treatment of coronary heart disease， and combined with TCM for relaxing tendons and activating collaterals， resolving phlegm to benefit orifices， and invigorating spleen to remove dampness combined for the treatment of lower extremity sclerosis. In addition， the medication time， drug combination and improvement indexes were summarized. In basic studies， the experimental progress of this kind of medicine for the treatment of AS were summed up in the aspect of reducing inflammatory reaction， improving the abnormal lipid metabolism and improving the damage of inner membrane. At present， it was found that tanshinone， total saponins of stem and leaf of Panax Quinquefolium， extract of Trichosanthis Pericarpium. Qishen Yiqi dropping pill， Huxinkang tablet， Danlou tablet， Buyang Huanwutang combined with Gualou Xiebaitang， Huazhuo Tongmai powder were the main drugs for basic research， and the animal model， model characteristics and the mechanism of action were summarized. In order to provide a reference for the rational application of TCM for Yiqi Huoxue Huatan in the treatment of AS， the application law， the mechanism and characteristics of action and the future research directions of TCM for Yiqi Huoxue Huatan were summarized and reviewed.

Mechanism of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma Extracts in Delaying High Glucose-induced Vascular Calcification in Mice / 中国实验方剂学杂志

Objective::To investigate the protective effect of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts on vascular calcification induced by high glucose in mice by observing the expression of osteopontin (OPN) and smooth muscle 22α (SM22α) as well as vascular calcium deposition in the common carotid artery and thoracic aorta of mice. Method::Totally 130 male C57BL/6 mice were randomly divided into normal control group and high glucose group. The mice in high glucose group were intraperitoneally injected with streptozotocin(STZ), and fed on a high-fat diet for 7 months. Then, the mice were randomly divided into model group, low-dose and high-dose Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts groups (0.819, 1.638 g·kg-1), and metformin group (150 mg·kg-1). Each group was intragastrically administered once a day for 9 weeks. The changes in blood glucose were measured. Seven days before the end of the administration, a group of 4-week old male C57BL/6 mice were purchased and fed normally for one week as a youth group. At the end of the administration, the common carotid artery and thoracic aorta tissues of the mice were collected. Von Kossa staining was used to determine the degree of calcium deposition in the common carotid artery and thoracic aorta. The expression levels of OPN and SM22α protein in the common carotid artery and thoracic aorta were detected by immunohistochemistry. The expression of OPN and SM22α protein in the common carotid artery of mice was determined by Western blot. Result::As compared with the young group, the blood glucose of the normal control group was slightly increased without statistical difference, the common carotid artery and thoracic aorta were uniformly stained, and no black granular precipitate was observed. As compared with the normal control group, the blood glucose of the model group was increased (P<0.01), with a large amount of brown-black particles deposited in the intimal elastic fibers, showing obvious calcium salt deposition. As compared with the model group, blood glucose was significantly decreased in each administration group (P<0.05, P<0.01), and the degree of vascular calcium salt deposition was significantly reduced. There were no significant changes in expression levels of OPN protein and SM22α protein in the common carotid artery and thoracic aorta between the youth group and normal control group. As compared with the normal control group, the expression of intimal OPN protein in the common carotid artery and thoracic aorta of the model group was positive, SM22α protein expression was weakly positive, and the gray value of OPN protein expression in the common carotid artery was significantly increased (P<0.01), while the gray value of SM22α protein was decreased significantly (P<0.01). As compared with the model group, the expression levels of intimal OPN protein and SM22α protein in the common carotid artery and thoracic aorta of each administration group were significantly improved, and the gray value of OPN protein expression in the common carotid artery was reduced (P<0.05, P<0.01), while SM22α protein expression was significantly increased (P<0.01). Conclusion::High glucose can induce calcification of common carotid artery and thoracic aorta in mice and accelerate vascular aging. This formation process may be related to the expression of OPN and SM22α. Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts can reduce vascular calcification and delay vascular aging by regulating the expression of OPN and SM22α.

Explore Protective Mechanism of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma Extract on Cardiac Aging in Diabetic Mice Through AMPK/mTOR Pathway / 中国实验方剂学杂志

Objective::To explore the protective mechanism of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma (GNC) extracts on cardiac aging in diabetic mice by observing the activation of AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway, changes of cardiac pathomorphological and related senescent proteins. Method::C57BL/6 male mice, SPF level, were randomly divided into normal control group and high-glucose group. The mice in high-glucose group were intraperitoneally injected with streptozotocin (STZ) and fed with high-fat diet. After successful modeling, they were randomly divided into model group, low-dose GNC group (0.819 g·kg-1), high-dose GNC group (1.638 g·kg-1) and metformin group (150 mg·kg-1). The drug was administered by gavage once a day for a continuous period of 9 weeks. 4-week-old male C57BL/6 mice were normally fed for 1 week as a youth group. General conditions of mice were observed. Hematoxylin-eosin (HE) staining combined with transmission electron microscope (TEM) was used to observe the cardiac pathomorphology in mice. Von Kossa staining was used to determine the degree of calcium salt deposition in cardiac micro vessels. Western blot was used to detect the activation of signaling pathways in myocardial tissue of mice, as well as the expression levels of matrix metalloproteinases-2 (MMP-2), tumor suppressor p53 (p53), and phospho-tumor suppressor p53 (p-p53). Result::As compared with the normal group, the blood glucose in the model group increased (P<0.01), as compared with the model group, the blood glucose in each administration group decreased significantly (P<0.05, P<0.01). The results of three pathological morphology experiments (HE, TEM, and Von Kossa) showed that as compared with the normal control group, the mice in model group showed cardiomyocytes hypertrophy, disordered arrangement of myocardial fibers, focal dissolving and necrosis, mitochondria swelling, degeneration, crest fracture, vacuolar alteration, disordered microvascular structure of the heart, uneven staining, and a large amount of calcium deposition in tunica media and intima. As compared with the model group, the pathomorphological changes of mice in each administration group were improved in varying degrees. Compared with the normal group, the expression levels of MMP-2, p53 and p-p53 protein in the model group were significantly increased (P<0.05, P<0.01), the protein ratios of p-liver kinase B2(LKB1)/LKB1, p-AMPK/AMPK were significantly decreased (P<0.05, P<0.01), and the average gray level of p-mTOR/mTOR and p-p70S6 kinase(p70S6k)/p70S6k protein was significantly increased (P<0.05, P<0.01), while the protein ratios of p-mTOR/mTOR, p-p70S6k/p70S6k were increased (P<0.01). As compared with the model group, the expression levels of MMP-2, p53 and p-p53 protein in each administration group were significantly decreased (P<0.05, P<0.01), the protein ratios of p-LKB1/ LKB1, p-AMPK/AMPK were significantly increased (P<0.05, P<0.01), while the protein ratios of p-mTOR/mTOR and p-p70S6k/p70S6k were decreased (P<0.05, P<0.01). Conclusion::STZ combined with high-fat diet can induce cardiac aging in mice, and GNC can improve cardiac aging in diabetic mice, which may be related to the inhibition of AMPK/mTOR pathway related protein expression.

Effect of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts on intestinal flora of vascular aging mice induced by high glucose and high lipid / 中国中药杂志

The aim of this paper was to observe the changes of intestinal flora in vascular aging mice, in order to explore the relationship between vascular aging and intestinal flora and the effects of extracts of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma on intestinal flora of vascular aging mice. A model of vascular aging in mice was induced through intrape-ritoneal injection with streptozotocin(STZ) combined with high-fat diet. Biochemical detection was performed on serum cholesterol(CHO), triglyceride(TG), high-density liptein cholesterol(HDL-C), low-density liptein cholesterol(LDL-C) and blood glucose(GLU). HE staining was used to detect mice thoracic aorta morphology, and the expressions of cyclin-dependent kinase inhibitor 2 A(p16) and cyclin-dependent kinase inhibitor 1 A(p21) protein in mice thoracic aorta were detected by Western blot. The 16 S rDNA gene of mice intestinal flora was detected by Illumina MiSeq high-throughput sequencing technology to explore the changes of intestinal flora in each group. The results demonstrated that the GLU level in low-dose and high-dose TCM groups decreased, but with unobvious changes in blood lipid indexes. Metformin could significantly decrease the levels of GLU(P<0.01), CHO and LDL-C in mice(P<0.05). Intravascular injury was not obvious in each drug group, and the expressions of p16 and p21 protein were significantly decreased(P<0.05). The intestinal flora of each group was mainly composed of Firmicutes(F) and Bacteroidetes(B) at the level of the phylum, but the B/F ratio was different from that of the youth group and the blank control group. The B/F ratio of the model group was significantly lower(P<0.01), and compared with the model group, the B/F ratio of the high-dose group and the metformin group was signi-ficantly higher(P<0.05). There were dominant and differential floras in the intestine of each group of mice. The results showed that extracts of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma could improve the intestinal flora structure and create a good intestinal environment by increasing the B/F ratio, which provides a new possible pathway for lowering blood glucose and blood lipids and delaying vascular aging.

Controlled Release of Naringin in GelMA-Incorporated Rutile Nanorod Films to Regulate Osteogenic Differentiation of Mesenchymal Stem Cells.

Naringin, a Chinese herbal medicine, has been demonstrated to concentration-dependently promote osteogenic differentiation of mesenchymal stem cells (MSCs). However, it remains a challenge to load naringin on coatings for osteogenesis and further control the release kinetics. Here, we demonstrated that the release behavior of naringin on rutile nanorod films could be controlled by either mixing naringin with gelatin methacryloyl (GelMA) before spinning onto the films or soaking the obtained GelMA-incorporated films with the naringin solution to achieve the distinct degradation-type release and diffusion-type release, respectively. We further revealed that the naringin-loaded coatings facilitated adhesion, proliferation and late differentiation, and mineralization of MSCs. Our findings provided a novel strategy to engineer the coatings with controlled release of naringin and emphasized the bioactivity of naringin for the osteogenic differentiation of MSCs.

Structure-activity relationship study and biological evaluation of SAC-Garlic acid conjugates as novel anti-inflammatory agents.

A series of S-allyl-l-cysteine (SAC) with garlic acid conjugates as anti-inflammatory agents were designed and synthesized. Among the 40 tested compounds, SMU-8c exhibited the most potent inhibitory activity to Pam3CSK4-induced nitric oxide (NO) in RAW264.7 macrophages with IC50 of 22.54 ±â€¯2.60 µM. The structure-activity relationship (SAR) study suggested that the esterified carboxyl group, carbon chain extension and methoxylation phenol hydroxy could improve the anti-inflammatory efficacy. Preliminary anti-inflammatory mechanism studies showed that SMU-8c significantly down-regulated the levels of Pam3CSK4 triggered TNF-α cytokine in human THP-1 cells, mouse RAW 264.7 macrophages, as well as in ex-vivo human peripheral blood mononuclear cells (PBMC) with no influence on cell viability. SMU-8c specifically blocked the Pam3CSK4 ignited secreted embryonic alkaline phosphatase (SEAP) signaling with no influence to Poly I:C or LPS triggered TLR3 or TLR4 signaling. Moreover, SMU-8c suppressed TLR2 in HEK-Blue hTLR2 cells and inhibited the formation of TLR1-TLR2, and TLR2-TLR6 complex in human PBMC. In summary, SMU-8c inhibited the TLR2 signaling pathway to down-regulate the inflammation cytokines, such as NO, SEAP and TNF-α, to realize its anti-inflammatory activity.

Interventional Effect and Mechanism of Extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma on Vascular Endothelial Cellular Senescence Induced by High Glucose and High Fat / 中国实验方剂学杂志

Objective: To explore the mechanism of extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma in delaying the senescence of vascular endothelial cells induced by high glucose and high fat. Method: The 40 mmol·L-1 glucose and 100 μmol·L-1 palmitate were used to induce endothelial cell senescence. The experiment was divided into control group, model group and low, medium and high-dose traditional Chinese medicine groups (50,100,200 mg·L-1). The intervention lasted for 48 h. Cell proliferation was detected by cell counting kit-8(CCK-8); cell senescence was detected by senescence β-galactosidase staining; p16 and p21 protein expression levels were detected by Western blot; p-H2A. X(Ser139) expression, mitochondria ROS(mtROS) production and changes in mitochondrial membrane potential(MMP) were detected by immunofluorescence. Result: Compared with the control group, in model group, the cell proliferation ability and the number of SA-β-gal blue-stained cells decreased(PPPPβ-gal blue-stained cells, the mtROS production, and expression levels of p16, p21 and p-H2A. X(Ser139)(PPConclusion: The above results suggest that extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma delay of endothelial cellular senescence induced by high glucose and high fat, and its mechanism may be related to increasing mitochondrial membrane potential and reducing DNA damage accumulation caused by ROS production.

[Effect of microRNA-34a/SIRT1/p53 signal pathway on notoginsenoside R1 delaying vascular endothelial cell senescence].

This study aimed to investigate the effect of notoginsenoside R1 in delaying H2O2-induced vascular endothelial cell senescence through microRNA-34a/SIRT1/p53 signal pathway. In this study， human umbilical vein endothelial cells(HUVECs) were selected as the study object; the aging model induced by hydrogen peroxide(H2O2) was established， with resveratrol as the positive drug. HUVECs were randomly divided into four groups， youth group， senescence model group， notoginsenoside R1 group and resveratrol group. Notoginsenoside R1 group and resveratrol group were modeled with 100 µmoL·L⁻¹ H2O2 for 4 h after 24 h treatment with notoginsenoside R1(30 µmoL·L⁻¹) and resveratrol(10 µmoL·L⁻¹) respectively. At the end， each group was cultured with complete medium for 24 h. The degree of cellular senescence was detected by senescence-associated ß-galactosidase(SA-ß-Gal) staining kit， the cell viability was detected by cell counting kit-8， the cell cycle distribution was analyzed by flow cytometry， and the cellular SOD activity was detected by WST-1 method in each group. The expressions of SIRT1， p53， p21 and p16 proteins in HUVECs were detected by Western blot. In addition， the mRNA expressions of miRNA-34a， SIRT1 and p53 in HUVECs were assayed by Real-time PCR. These results indicated that notoginsenoside R1 significantly reduced the positive staining rate of senescent cells， enhanced the cell proliferation capacity and intracellular SOD activity， decreased the proportion of cells in G0/G1 phase， and increased the percentage of cells in S phase simultaneously compared with the senescence model group. Moreover， notoginsenoside R1 decreased the mRNA expressions of miRNA-34a and p53 and the protein expression of p53， p21 and p16.At the same time， notoginsenoside R1 increased the protein and mRNA expressions of SIRT1. The differences in these results between the senescence model group and the notoginsenoside R1 group were statistically significant(P<0.05). However， there was not statistically significant difference in these results between the notoginsenoside R1 group and the resveratrol group. In conclusion， the senescence of endothelial cells induced by H2O2 can be used as a model for studying aging. Notoginsenoside R1 has an obvious anti-aging effect on vascular endothelial cells in this study. The possible mechanism is that notoginsenoside R1 can delay the senescence process of vascular endothelial cells induced by H2O2 by regulating microRNA-34a/SIRT1/p53 signal pathway.

Controlled Release of Naringin in Metal-Organic Framework-Loaded Mineralized Collagen Coating to Simultaneously Enhance Osseointegration and Antibacterial Activity.

Two important goals in orthopedic implant research are to promote osseointegration and prevent infection. However, much previous effort has been focused on the design of coatings to either enhance osseointegration while ignoring antibacterial activity or vice versa, to prevent infection while ignoring bone integration. Here, we designed a multifunctional mineralized collagen coating on titanium with the aid of metal-organic framework (MOF) nanocrystals to control the release of naringin, a Chinese herbal medicine that could promote osseointegration and prevent bacterial infection. The attachment, proliferation, osteogenic differentiation, and mineralization of mesenchymal stem cells on the coating were significantly enhanced. Meanwhile, the antibacterial abilities against Staphylococcus aureus were also promoted. Furthermore, release kinetics analysis indicated that the synergistic effect of a primary burst release stage and secondary slow release stage played a critical role in the performance and could be controlled by the relative concentrations of MOF and naringin. This work thus provides a novel strategy to engineer multifunctional orthopedic coatings that can enhance osseointegration and simultaneously inhibit microbial cell growth.