Visual tracking of label-free microplastics in wheat seedlings and their effects on crop growth and physiology.

The effects of microplastics on crop plants have attracted growing attention. However, little is known about the effects of microplastics and their extracts on the growth and physiology of wheat seedlings. In this study, hyperspectral-enhanced dark field microscopy and scanning electron microscopy were used to accurately track the accumulation of 200 nm label-free polystyrene microplastics (PS) in wheat seedlings. The PS accumulated along the root xylem cell wall and in the xylem vessel member and then moved toward to the shoots. In addition, lower concentration (≤ 5 mg·L-1) of microplastics increased root hydraulic conductivity by 80.6 %- 117.0 %. While higher PS treatment (200 mg·L-1) considerably decreased plant pigments content (chlorophyll a, b, and total chlorophyll) by 14.8 %, 19.9 %, and 17.2 %, respectively, and decreased root hydraulic conductivity by 50.7 %. Similarly, catalase activity was reduced by 17.7 % in root and 36.8 % in shoot. However, extracts from the PS solution showed no physiological effect on wheat. The result confirmed that it was the plastic particle, rather than the chemical reagents added in the microplastics, contributed to the physiological variation. These data will benefit to better understanding on the behavior of microplastics in soil plants, and to providing of convincing evidence for the effects of terrestrial microplastics.

Purple Rehmannia: Investigation of the activation of R2R3-MYB transcription factors involved in anthocyanin biosynthesis.

Engineering anthocyanin biosynthesis in herbs could provide health-promoting foods for improving human health. Rehmannia glutinosa is a popular medicinal herb in Asia, and was a health food for the emperors of the Han Dynasty (59 B.C.). In this study, we revealed the differences in anthocyanin composition and content between three Rehmannia species. On the 250, 235 and 206 identified MYBs in the respective species, six could regulate anthocyanin biosynthesis by activating the ANTHOCYANIDIN SYNTHASE (ANS) gene expression. Permanent overexpression of the Rehmannia MYB genes in tobacco strongly promoted anthocyanin content and expression levels of NtANS and other genes. A red appearance of leaves and tuberous/roots was observed, and the total anthocyanin content and the cyanidin-3-O-glucoside content were significantly higher in the lines overexpressing RgMYB41, RgMYB42, and RgMYB43 from R. glutinosa, as well as RcMYB1 and RcMYB3 in R. chingii and RhMYB1 from R. henryi plants. Knocking out of RcMYB3 by CRISPR/Cas9 gene editing resulted in the discoloration of the R. chingii corolla lobes, and decreased the content of anthocyanin. R. glutinosa overexpressing RcMYB3 displayed a distinct purple color in the whole plants, and the antioxidant activity of the transgenic plants was significantly enhanced compared to WT. These results indicate that Rehmannia MYBs can be used to engineer anthocyanin biosynthesis in herbs to improve their additional value, such as increased antioxidant contents.

Sequential washing and eluent regeneration with agricultural waste extracts and residues for facile remediation of meta-contaminated agricultural soils.

Washing with organic acids and dissolved organic carbon (DOC) is a promising technique for effective removal of potentially toxic metals from agricultural soils and the two key factors are the screening of inexpensive, high-efficiency, and environmentally friendly washing agents and the safe treatment of waste eluent. We used extracts from agro-forestry wastes (pineapple peel, lemon peel, grapefruit peel and gardening crabapple fruit) to develop a facile two-stage sequential washing method (extracts and/or citric acid (CA) and coupled with extracts) and regenerated waste eluent. The washing efficiencies of Cd and Cu were significantly increased by pineapple peel (PP) using two-stage sequential washing with the sequence of PP + CA-PP > CA-PP > PP-PP. The potential pollution risk from soil Cd was lowered by 33.0% from moderate to low risk, and soil nutrient contents increased. 80.9% of Cd and 81.3% of Cu in waste eluent were efficiently removed by the PP residues. The removal mechanisms of metals in soils and eluents by PP washing agents and residues can be attributed to acid activation, cation exchange and complexation between metal ions and carboxyl groups. Therefore, the PP extracts and residues are potentially suitable for the removal of Cd and Cu from polluted agricultural soils and washing waste eluents.

[A theoretical and experimental study on the Fuzzy evaluation model of biological performance of Chinese materia medica with an either cold or hot herbal property].

OBJECTIVE: The concepts of cold and hot herbal properties in traditional Chinese medicine (TCM), reflecting the efficacy trends of medicinal herbs, have the Fuzzy characteristics. It is suited for the application of Fuzzy mathematics in research of herbal properties. For a comprehensive evaluation of herbal properties, this work constructed and made a practice of the Fuzzy discrimination model regarding the biological performance of Chinese materia medica. METHODS: Firstly, TCM formulas of cold or hot property were employed for setting the reference values and calculating the membership functions of cold and hot Fuzzy sets with Fisher discriminant. Then the model parameters of cold performance function (CPF) and hot performance function (HPF) were acquired. Second, the experimental data from studies on changes in multisystem indicators of animals given the classical cold formula (Sanhuang Fang) and hot formula (Jiangfugui Fang) were collected to calculate CPF and HPF. Then an experiment of administration of hot property herbs, Rougui (Cortex Cinnamomi) and Xianmao (Rhizoma Curculiginis), and cold property herbs, Huangbai (Cortex Phellodendri) and Zhizi (Fructus Gardeniae) to normal animals and animals with glucocorticoid-induced yang-deficiency or yin-deficiency state was designed. The relative changes in animal states were analyzed by the evaluation model to find the way in which herbal properties are represented. RESULTS: The HPF values of biological performance were higher than the CPF values in administration of Rougui to animals of normal (0.528>0.221), yang-deficiency (0.203>-0.490) and yin-deficiency (1.750>-0.479) states, and in administration of Xianmao to animals of normal (0.474>-1.601) and yang-deficiency (0.288>-1.923) states, which represented the hot property. The HPF values of biological performance were lower than the CPF values in administration of Huangbai to animals of normal (-0.870<0.798) and yang-deficiency (-0.339<0.194) states, and in administration of Zhizi to animals of yin-deficiency (-1.802<0.354) state, which represented the cold property. CONCLUSION: The herbal properties and the biological performance characteristics of herbal formulas are ascertained by using the Fuzzy evaluation model. The personalities, as well as commonalities of herbal properties are key areas for focus because of the multi-approach biological performance of cold and hot property.

[Ridit analysis of experimental data from animal models of yang deficiency induced by different doses of hydrocortisone].

OBJECTIVE: Hydrocortisone-induced yang-deficiency animal model has now become the generally accepted model of yang deficiency. However, assessing the most appropriate dose of hydrocortisone is a long-term challenge. For analyzing the modeling dose, the authors have built several kinds of yang-deficiency models induced by hydrocortisone at different doses, and analyzed the experimental data with various mathematical statistical methods. In order to discuss the effects of the modeling dose on the basis of previous research, the authors introduced Ridit analysis. METHODS: After categorizing 27 batches of experiments with four different doses (including 2.5, 3.75, 10 and 20 mg/kg), the data were standardized and the data type was changed to meet the needs of subsequent calculations. Then, by using Ridit analysis, the authors compared the variation of 19 biochemistry indexes involving nervous-endocrine system, immune system, metabolic system and the function of the liver and kidney, so as to analyze the similarities and differences of those yang-deficiency models mentioned above. The effects of modeling dose were then discussed. RESULTS: With regard to the overall state of the animal model, the difference between the models induced by different doses of hydrocortisone (2.5, 3.75, 10 and 20 mg/kg) showed no statistical significance, indicating that differences in hydrocortisone dose barely lead to significant changes in yang-deficiency models. For the trends of changing in different indexes, indexes had different performances when the doses differs: gonad indexes (estradiol and testosterone) showed better performance in 2.5 mg/kg group, immunological indexes (immunoglobulin M and immunoglobulin G) showed better performance in 3.75 mg/kg group and metabolic indexes (triglyceride and total cholesterol) showed better performance in 10 mg/kg group, etc. This indicates that the dose of hydrocortisone should be determined according to the research purposes. CONCLUSION: Ridit analysis can be used as an integration analysis method for animal models of yang deficiency induced by hydrocortisone at different doses.

[Pharmacological effects of Oleum Cinnamomi and water extract of Cortex Cinnamomi in rats with yang-deficiency cold syndrome and the mathematical analysis].

OBJECTIVE: To compare the effects of Oleum Cinnamomi and water extract of Cortex Cinnamomi in rats with yang-deficiency cold syndrome based on mathematical analysis. METHODS: A total of 48 male Sprague-Dawley rats were randomly divided into 4 groups (normal, model, Oleum Cinnamomi and water extract of Cortex Cinnamomi). Yang-deficiency cold syndrome was induced by hydrocortisone sodium succinate. Oleum Cinnamomi and water extract of Cortex Cinnamomi were given by gastrogavage daily for one week to respective groups. Material metabolism indexes such as glucose (GLU), uric acid (UA), total cholesterol (TC), triacylylyceral (TAG), total protein (TP) and albumin (ALB); energy metabolism indexes such as lactic acid (LAC), lactate dehydrogenase (LDH), succinate dehydrogenase (SDH) and adenosine triphosphatase (ATPase); endocrine system indexes such as corticosteroid (CS), triiodothyronine (T3), tetraiodothyronine (T4), thyroid stimulating hormone (TSH), estradiol (E(2)), vitamin C (VC) and 17-hydroxycorticosteriod (17-OHCS); and immune system indexes such as immunoglobulin M (IgM), immunoglobulin G (IgG), complement 3 (C3) and complement 4 (C4) were measured. And then the data were analyzed by mathematical analysis method. RESULTS: Oleum Cinnamomi and water extract of Cortex Cinnamomi had similar influence on some indexes of material metabolism, energy metabolism and endocrine and immune systems in rats with yang-deficiency cold syndrome. Positive effects were showed in GLU, TC, TAG, TP, ALB, IgM, E(2), CS, VC and 17-OHCS. Water extract of Cortex Cinnamomi had no significant effects on TSH, LAC and SDH, but Oleum Cinnamomi had, while water extract of Cortex Cinnamomi had significant effects on ATPase, LDH and IgG. CONCLUSION: There is a complex relationship between Oleum Cinnamomi and water extract of Cortex Cinnamomi in their effects on rats with yang-deficiency cold syndrome. They have similar property, however, disparities exist between them to some extent. The exact mechanism needs further research.

[Study of biological performance of Chinese materia medica with either a cold or hot property based on the three-element mathematical analysis model].

OBJECTIVE: The properties of Chinese materia medica are believed to be the summarization of the effects of biological performance on the various body states. Systemic discussion of chemical-factor elements, body-condition elements, biological-performance elements and their interrelationships is needed for research into the properties of Chinese materia medica. Following the practical characteristics of Chinese medicine, the three-element mathematical model was formed by introducing some mathematical concepts and methods and was used to study the cold or hot property of Chinese medicine, and to investigate the difference in biological performances of the two properties. METHODS: By using the concept of different functionality of Chinese medicine on abnormal states and the idea of interaction in mathematics, the effects of chemical-factor elements and body-condition elements were normalized to the amount of biological performance which was represented by some important indicators. The three-element mathematical model was formed with scatter plots through four steps, including effect separation, intensity calculation, frequency statistics and relevance analysis. A comparison pharmacology experiment of administration of hot property medicines, Fuzi (Radix Aconiti Lateralis Preparata) and Rougui (Cortex Cinnamomi), and cold property medicines, Huangbai (Cortex Phellodendri) and Zhizi (Fructus Gardeniae) on normal and glucocorticoid-induced yang-deficiency and yin-deficiency states was designed. The results were analyzed by the mathematical model. The scatter plots were the main output of model analysis. The expression of cold property and hot property was able to be quantified by frequency distribution of biological indexes of administrations on yang-deficiency and yin-deficiency states in the "efficacy zone" and "toxicity zone" of the plots and by the relevance analysis. RESULTS: The ratios of biological indicator frequency in the "efficacy zone" of administrations on yang-deficiency state and yin-deficiency state were 7:3 for Fuzi, 3:3 for Rougui, 4:4 for Huangbai and 1:5 for Zhizi. The sums of the biological indicator frequency in the "toxicity zone" of administration on the two states were 4 for Fuzi, 0 for Rougui, 2 for Huangbai and 4 for Zhizi. The relevance analysis showed that the order from Fuzi, Rougui, Huangbai to Zhizi was proportional to the change from "be true of yang-deficiency state" to "be true of yin-deficiency state". The extent of the hot property decreased while that of the cold property increased in the order of Fuzi, Rougui, Huangbai and Zhizi. The stronger the efficacy of above medicines is, the more obvious the toxicity displayed. CONCLUSION: The three-element mathematical model employed in this study is effectively capable of explaining the different biological expressions between hot property medicines and cold property medicines. This suggests that it may provide a mathematical tool and theoretical basis for the modern interpretation of cold property and hot property of Chinese medicine, and provide new ideas for further studing into the essence of Chinese medicine property theory.

[Mathematical analysis of characteristics of glucocorticoid-induced yang deficiency or yin deficiency syndrome in animal models based on information entropy theory].

OBJECTIVE: Based on information entropy theory, this study analyzes the experimental indicators of glucocorticoid-induced traditional Chinese medicine yang deficiency or yin deficiency syndrome in animal models, thus laying the foundation of defining and interpreting the model state. METHODS: Data of biochemical indicators from 24 groups of animal models with different modeling conditions (type of modeling agent, dosage of modeling agent and modeling time) were collected. Information gain values for three study objects (type of modeling agent, dosage of modeling agent and modeling time) were calculated respectively after standardization, and then characteristics of yang deficiency or yin deficiency syndrome models were interpreted with these values and ranking map. RESULTS: Greatest information gain values of most indicators were got when the study object was the type of modeling agent, which is the most important factor in the differentiation of model state. With this study object, triglyceride (TG), total cholesterol (TC), total protein (TP), albumin (ALB), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and corticosteroid (CS) got larger information gain values than testosterone (T), lactic dehydrogenase (LDH), and cyclic adenosine monophosphate (cAMP). This indicated that the former seven indicators may be significantly different between the two animal models induced by hydrocortisone and dexamethasone, respectively. In the study of the modeling time, AST may be affected more than others. In the study of the dosage of modeling agent, TC and ALB may be affected more in the model of hydrocortisone and no indicators were significantly affected by the dosage of modeling agent in the dexamethasone model. CONCLUSION: Mathematical method based on information entropy theory allows researchers to analyze experimental data in several experiments at the same time and to interpret the trend and stability of indicators. Based on this study and by analyzing the impacts of different modeling conditions on the indicators, elucidations of the characteristics of glucocorticoid-induced yang deficiency or yin deficiency syndrome in animal models and some biochemical indicators for model comparison are in need.