Relationship between the UPLC Fingerprints of Citrus reticulata "Chachi" Leaves and Their Antioxidant Activities.

Citrus reticulata "Chachi" (CRC) leaves contain abundant flavonoids, indicating that they possess good nutritional/pharmacological research and development potential. This study aims to explore chemical antioxidant quality markers based on the spectrum-effect relationship and quality control strategy of CRC leaves. The ultrahigh performance liquid chromatography (UPLC) system was used to establish chromatographic fingerprints of Citrus reticulata "Chachi" leaves. Simultaneously, they were evaluated by using similarity analysis (SA), hierarchical cluster analysis (HCA), and principal component analysis (PCA). Afterwards, the DPPH assay was adopted to study the antioxidant effects. The spectrum-effect relationship between UPLC fingerprints and DPPH radical-scavenging activities was studied with grey relational analysis (GRA). Analysis results indicated that there were twenty-one common peaks of fourteen batches of CRC leaves which were from different regions of Guangdong province, and their similarities ranged from 0.648 to 0.997. HCA results showed that fourteen batches of samples of CRC leaves could be divided into six classes at Euclidean distance of 5. The results from GRA showed that tangeretin and hesperidin were the main flavonoids responsible for the antioxidant activity in CRC leaves. In conclusion, this research established a chromatographic analysis method suitable for CRC leaves and demonstrated that chromatographic fingerprints analysis combined with the antioxidant activity could be used to evaluate the material basis of CRC leaves and may provide a reference to establish a quality standard.

Metal-free visible-light-driven cascade cyclization reaction to synthesize 2-oxindoles via benzoyl and phenylsulfinyl radicals with acrylamide derivatives.

A metal-free visible-light-driven cascade cyclization reaction to synthesize 3-methyl-3-acetophenone-2-oxindoles and 3-methyl-3-(methylsulfonyl)benzene-2-oxindoles in yields up to 96% and 99%, via benzoyl and phenylsulfinyl radicals with acrylamide derivatives is reported, respectively. Extensive studies, including gram-scale, radical capture and isotope experiments, were performed to indicate that the reaction may involve a radical process.

[Current status and development strategy of ISO/TC 249 standards for Chinese materia medica].

The internationalization of traditional Chinese medicine(TCM) is one of the strategic development objectives in China, which has been incorporated into the national strategy as an important part of the Belt and Road Initiative development strategy. As the basis and prerequisite of TCM development, Chinese materia medica(CMM) has a direct impact on the internationalization of TCM. The International Organization for Standardization(ISO) is a global organization composed of national standardization bodies, and the ISO standards impact the world's economy, trade, communication and cooperation. Based on a brief introduction to ISO/Traditional Chinese Medicine Technical Committee(ISO/TC 249), this study elaborates the necessity of establishing ISO standards for CMM and analyzes the current status and challenges faced by the formulation of international standards for CMM. Finally, this study puts forward the development strategy of international standards for CMM. Specifically, efforts should be made to develop top-level design with international market demands as the guidance and improve the quality of standards to accelerate the transformation of domestic high-quality standards into international standards. Moreover, measures should be taken to give full play to the positive role of enterprises in the formulation of standards, vigorously cultivate compound talents for international standardization of TCM, and constantly strengthen international cooperation. The experience and thinking are of guiding significance for the scientific, efficient and reasonable formulation of high-quality ISO standards for CMM in the future.

[Analysis of economic benefits of Chinese medicine eco-agriculture based on multiple stakeholders].

As the most advanced environment-friendly production model in the international society, ecological agriculture of Chinese materia medica(CMM) is the only way for the development of modern agriculture. With the proposal of the declaration on ecolo-gical agriculture of CMM, "Don't grab land from farmland, don't be enemies of grass and insects, don't be afraid of barren slopes and forests, and live up to the green and green mountains", the ecological planting of CMM has blossomed all over the country, and formed a scientific theory, technology and model. Based on the theory and method of economics, this paper expounds the comprehensive benefits and development advantages of ecological agriculture of CMM from the perspectives of farmers(producers), patients(consumers) and the country. From the perspective of medicinal farmers, the input and output income of conventional agriculture and ecological agriculture of CMM such as Panax ginseng, Astragalus propinquus, Atractylodes lancea, and Bupleurum chinense were compared, and it was found that ecological agriculture of CMM had obvious advantages in net income, average annual income and input-output ratio, which could better promote farmers' income. From the perspective of patients, according to the same dose, the content of active ingredients in ecologically planted CMMs is significantly higher than that in conventionally-planted herbs, and the amount of effective substances taken by patients is also higher, so as to achieve better therapeutic effect. At the national level, ecological planting of CMM is the key to ensuring the high-quality development of CMM industry, increasing farmers' income, ensuring the safety of people's drug use and promoting the sustainable development of agriculture. It is also an important part of realizing the harmonious development of economy, society and environment and promoting ecological civilization. In general, the declaration on ecological agriculture of CMM embo-dies the core characteristics and goals of ecological agriculture, and also points of the path and vision of ecological agriculture of CMM in the future. The declaration will guide production practice, promote the benefit of farmers, and lay the foundation for the sustainable development of CMM industry.

[Discussion on high quality development and authentication system construction of Dao-di herbs based on blockchain].

Dao-di herbs are an important part of clinical medicine in traditional Chinese medicine. They are also precious wealth left to human beings from history, which contain deep traditional Chinese cultural connotations and play an important role in supporting and serving the Chinese medicine business. The relevant policy documents introduced by various national ministries and commissions have many contents and requirements related to the promotion of Dao-di herbs protection and industrial development. Due to the Dao-di herbs industry has a series of characteristics, such as a long chain, many involved links, long cycles, multiple production entities, multiple locations, and various types, the high-quality development of the industry has put forward higher requirements on the linkage between upstream and downstream, production entities, traceability of the whole process and information sharing. This article takes Dao-di herbs certification work as an application scenario and entry point, and discusses it from the perspective of block chain and information technology. It proposes the following work ideas: establish multi-party consensus from the macro-organizational management, business, and operational technical levels, and unblock channels for data and information, to achieve institutionalization of certification; establish certification-related standards and specifications to achieve certification standardization; build a certification hardware system to achieve certification networking; build a certification software system to develop functions for specific information content such as identity, origin, production, production process, quality, product and brand of authentic medicinal material production interactively, and realize certification programmatic; data security and sharing of related production activities to achieve socialization of certification. Make full use of modern technologies such as blockchain, the internet of things, big data and information technology, and through the joint participation of management, production, use and the public, the whole process information of Daodi herbs is integrated to form an interconnected information sharing application mode, thus, to serve and promote the high-quality development of Dao-di herbs industry.

[Cultivation strategy on cluster brand of ecological agriculture of Dao-di herbs].

The cluster brand is the embodiment of the core competitiveness of an industry. Developing and cultivating cluster brand of ecological agriculture of Dao-di herbs not only helps to optimize the value chain of the Chinese medicinal materials(CMMs) industry cluster, realize the value-added of the CMMs industry cluster, but also enhance the visibility and influence of the industrial cluster, enhance the core competitiveness of the industrial cluster. This has important practical significance for promoting the "orderly" "safe" and "effective" development of the Dao-di herbs. Based on the industry development status of CMMs, this article introduces several concepts related to cluster brands and their relationships, and focuses on the cultivation models and strategies of cluster brand in the CMMs industry. Based on the current status of the development of the CMMs industry, this article introduces several concepts related to cluster brands and their interrelationships. It discusses the cultivation models and strategies of cluster brands in the CMMs industry, industry associations, Chinese medicine companies and individual growers as the support, insists on the ecological cultivation of authentic medicinal materials and the cultivation of cluster brands. Finally, it points out the direction for the high-quality development of the ecological agriculture of CMMs.

Structural characterization and discrimination of Ophiopogon japonicas (Liliaceae) from different geographical origins based on metabolite profiling analysis.

The root tuber of Ophiopogon japonicus (Thunb.) Ker-Gawl ("Maidong" in Chinese), with steroidal saponins and homoisoflavonoids as its representative chemical compositions, is a representative medicinal herbs with multiple major producing areas. This study aimed to distinguish the O. japonicas samples from Zhejiang and Sichuan by using an ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF MS)-based metabolome analysis. Firstly, a global chemical constituent identification of O. japonicas was carried out by using both automatic and manual methods. An integrated steroidal saponins structural identification strategy in O. japonicas based on exact mass information, fragmentation characteristics and retention time was developed. Overall, 135 steroidal saponins, 47 homoisoflavonoids and 9 other metabolites were quickly identified or tentatively identified from the MSE continuum data. Furthermore, multivariate statistical analysis revealed that O. japonicas from Zhejiang and Sichuan can clearly be separated and some markers were screened. Moreover, some major active components including total soluble sugar, total soluble polyphenol, total flavonoid, total saponin and 10 specific compounds were analyzed quantitatively. In general, these results showed that there were many differences between the metabolic profile data of O. japonicas from different producing areas, O. japonicas from Sichuan showed higher level steroidal saponins and samples from Zhejiang had higher contents of homoisoflavonoids specifically, and indicated that metabolite profiling by UPLC/Q-TOF MS is an effective approach for the discrimination of medicinal herbs from different geographical origins.

Chemical profiling in Moutan Cortex after sulfuring and desulfuring processes reveals further insights into the quality control of TCMs by nontargeted metabolomic analysis.

As a traditional processing method, sulfuring has been used in the processing of many traditional Chinese medicines (TCMs). Desulfuring, which has emerged in recent years, is a new method applied to sulfured herbs so they can comply with regulations regarding residual SO2. Due to the chemical transformations and the residual SO2 in the herbs, both sulfuring and desulfuring have negative effects on the safety and therapeutic effects of TCMs, and Moutan Cortex is one of the TCMs most susceptible to these effects. Here, a new strategy was developed to differentiate normal, sulfured and desulfured Moutan Cortex, and the transformations of compounds in sulfuring and desulfuring processes were analyzed using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MSE) method based on metabolomic analysis. Our findings were as follows: (1) a total of 119 compounds were identified or tentatively identified, including 9 compounds that are being reported for the first time as natural products; (2) 15 sulfocompounds were generated during the sulfuring process; (3) these sulfocompounds could not be converted back into their corresponding glycosides by the desulfuring process, and the desulfuring decreased the residual SO2,while also removing some soluble compounds in the sulfured Moutan Cortex; and (4) 28 compounds were screened and tentatively identified as markers for distinguishing normal, sulfured and desulfured Moutan Cortex. Our findings provide a new practical strategy for evaluating how sulfuring and desulfuring affect the quality of TCMs.

[Preparation and evaluation of gastric adhesive-floating pellets for Bolo leaf phenols].

Gastric adhesive-floating pellets for Bolo leaf phenols (BLP) were prepared by extrusion-spheronization method, with chitosan as skeleton bioadhesive material, and stearyl alcohol as help-bleaching agent to evaluate its in vitro adhesivity, floatability and in vivo retention situation, and investigate its in vitro release characteristics. The in vitro adhesivity and floatability were evaluated respectively by in vitro tissue retention method and visual observation method. The retention of pellets in rats was investigated by in vivo tissue retention method and in vivo imaging of small animals. In addition, the in vitro release of p-coumaric acid and caffeic acid as the index components in pellets were evaluated. Results showed that the in vitro adhesivity of the prepared gastric adhesive-floating pellets reached (73.2±3.4)%, and the pellets could float immediately in simulated gastric fluid for more than 12 h; the retention rate of adhesive-floating pellets in rats reached more than 40% after 6 h, while the retention rate of common reference pellets was decreased by 15% as compared with the gastric adhesive-floating pellets, with significant difference (P<0.01); the drug in vitro release time can reach more than 6 h, and the drug release behaviors were lined with Higuchi equation. In vivo and in vitro studies showed that, the gastrointestinal bioadhesive and floating pellets prepared in this study have good bioadhesivity, floatability and good sustained release characteristics.

[Bioavailability in vivo and in vitro transepithelial transport across Caco-2 cell monolayer of pueraria flavonoids bio-adhesive and floating pellets].

To study the bioavailability of pueraria flavonoids bio-adhesive and floating pellets, the absorption of puerarin was studied using Caco-2 cell monolayer by liquid chromatography (HPLC) method, comparing the P(app) of pueraria flavonoids bio-adhesive and floating pellets with different bio-adhesive materials. Drugs were administered at a dose of 100 mg·kg(-1) via ig. The plasma concentration of puerarin was determined by HPLC, the pharmacokinetics were calculated with the WinNonlin 6.0 software. The results showed that the P(app) of bio-adhesive and floating pellets with hydroxypropyl methylcellulose (HPMC)-cabomer was largest, which had a significant difference (P < 0.05). The AUC(0-t) of pueraria flavonoids bio-adhesive and floating pellets was 1.79 times of pueraria flavonoids, the C(max) of pueraria flavonoids bio-adhesive and floating pellets and pueraria flavonoids had a significant difference (P < 0.05). What's more the MRT had prolonged. In conclusion, pueraria flavonoids bio-adhesive and floating pellets with HPMC-cabomer could significantly facilitate the transport of puerarin on Caco-2 cellular monolayers. The bioavailability of pueraria flavonoids bio-adhesive and floating pellets with HPMC-cabomer was increased more than pueraria flavonoids with a sustained release effect.

[Discussion on present situation of study on pesticide residues in Chinese herbal medicines].

Pesticide residues in traditional Chinese medicine has attracted widespread attention at home and abroad. This paper analyzed the pollution present situation and existing problems of pesticide residue for Chinese herbal medicines, explicited the analytical methods of pesticide residues in Chinese herbal medicines. Meanwhile, the commonly used pesticide residue degradation and application in Chinese herbal medicines were discussed. Moreover, on the basis of analysis of pesticide residue standards, this paper proposed the necessity and urgency of the limit standard of pesticide residues in Chinese herbal medicines, and provided a scientific references for deepening research and developing safe, green medicines.

The Biosynthetic Pathways of Tanshinones and Phenolic Acids in Salvia miltiorrhiza.

Secondary metabolites from plants play key roles in human medicine and chemical industries. Due to limited accumulation of secondary metabolites in plants and their important roles, characterization of key enzymes involved in biosynthetic pathway will enable metabolic engineering or synthetic biology to improve or produce the compounds in plants or microorganisms, which provides an alternative for production of these valuable compounds. Salvia miltiorrhiza, containing tanshinones and phenolic acids as its active compounds, has been widely used for the treatment of cardiovascular and cerebrovascular diseases. The biosynthetic analysis of secondary metabolites in S. miltiorrhiza has made great progress due to the successful genetic transformation system, simplified hairy roots system, and high-throughput sequencing. The cloned genes in S. miltiorrhiza had provided references for functional characterization of the post-modification steps involved in biosynthesis of tanshinones and phenolic acids, and further utilization of these steps in metabolic engineering. The strategies used in these studies could provide solid foundation for elucidation of biosynthetic pathways of diterpenoids and phenolic acids in other species. The present review systematically summarizes recent advances in biosynthetic pathway analysis of tanshinones and phenolic acids as well as synthetic biology and metabolic engineering applications of the rate-limiting genes involved in the secondary metabolism in S. miltiorrhiza.

[Good agricultural practice (GAP) of Chinese materia medica (CMM) for ten years: achievements, problems and proposals].

This paper aims to summarize the achievements during the implementation process of good agricultural practice (GAP) in Chinese Materia Medica (CMM), and on basis of analyzing the existing problems of GAP, to propose further implementation of GAP in TCM growing. Since the launch of GAP in CMM growing ten years ago, it has acquired great achievements, including: (1) The promulgation of a series of measures for the administration of the GAP approval in the CMM growing; (2) The expanded planting area of CMM; (3) The increased awareness of standardized CMM growing among farmers and enterprises; (4) The establishment of GAP implementation bases for CMM growing; (5) The improvement of theory and methodology for CMM growing; (6) The development of a large group of experts and scholars in GAP approval for CMM production. The problems existing in the production include: (1) A deep understanding of GAP and its certification is still needed; (2) The distribution of the certification base is not reasonable; (3) The geo-economics effect and the backward farming practices are thought to be the bottlenecks in the standardization of CMM growing and the scale production of CMM; (4) Low comparative effectiveness limits the development of the GAP; (5) The base of breeding improved variety is blank; (6) The immature of the cultivation technique lead to the risk of production process; (7) The degradation of soil microbial and the continuous cropping obstacle restrict the sustainable development of the GAP base. To further promote the health and orderly GAP in the CMM growing, the authors propose: (1) To change the mode of production; (2) To establish a sound standard system so as to ensure quality products for fair prices; (3) To fully consider the geo-economic culture and vigorously promote the definite cultivating of traditional Chinese medicinal materials; (4) To strengthen the transformation and generalization of basic researches and achievements, in order to provide technical support for the CMM production; (5) To deepen the understanding of GAP, to vigorously promote ecological planting and precision agriculture, in order to overcome the continuous cropping obstacle. The authors think that despite the fact that we are still facing with a huge array of management and technological problems, the GAP in the CMM growing has already enjoyed widespread support and showed great potential. In the future, with people's deeper understanding of GAP and the great progress of the science and technology, the GAP will constantly be fused with the theory, methodology and technology in the modern agriculture like precision agriculture, eco-agriculture and etc.

Research advances on flotillins.

The proteins of flotillin-1 and flotillin-2 were originally discovered in axon regeneration of goldfish retinal ganglion cells. They are generally used as marker proteins of lipid rafts and considered to be scaffolding proteins of lipid microdomains. Although they are ubiquitously expressed and well-conserved from fly to man, their exact functions remain controversial. In this review, we summarize the structure of flotillins and some functions of them, such as regulating axon regeneration, endocytosis, T cell activation, insulin signaling, membrane protein recruitment, roles in the progression of some diseases and so on.

Mapping codon usage of the translation initiation region in porcine reproductive and respiratory syndrome virus genome.

BACKGROUND: Porcine reproductive and respitatory syndrome virus (PRRSV) is a recently emerged pathogen and severely affects swine populations worldwide. The replication of PRRSV is tightly controlled by viral gene expression and the codon usage of translation initiation region within each gene could potentially regulate the translation rate. Therefore, a better understanding of the codon usage pattern of the initiation translation region would shed light on the regulation of PRRSV gene expression. RESULTS: In this study, the codon usage in the translation initiation region and in the whole coding sequence was compared in PRRSV ORF1a and ORFs2-7. To investigate the potential role of codon usage in affecting the translation initiation rate, we established a codon usage model for PRRSV translation initiation region. We observed that some non-preferential codons are preferentially used in the translation initiation region in particular ORFs. Although some positions vary with codons, they intend to use codons with negative CUB. Furthermore, our model of codon usage showed that the conserved pattern of CUB is not directly consensus with the conserved sequence, but shaped under the translation selection. CONCLUSIONS: The non-variation pattern with negative CUB in the PRRSV translation initiation region scanned by ribosomes is considered the rate-limiting step in the translation process.