High-Quality Indigo Naturalis Obtained with Automatic Foam Separation.

Indigo Naturalis is not only an ancient plant dye but also a famous herbal medicine with antibacterial, anti-inflammatory, and anticancer properties. In traditional processes, thousands of manual stirring separate the high-quality Indigo Naturalis from the crude pulp system. However, this method is time-consuming and labor-intensive, resulting in an unstable quality and low yield, which cannot meet the requirements of modern industrial production. In this study, foam-separation technology was used to increase the industrial applicability of high-quality Indigo Naturalis. The process parameters were optimized based on the content of active ingredients, skin irritation effects, and antioxidative stress activity. The results showed that the optimal process of the foam separation achieved the liquid level difference of 40 cm and the foaming intensity of 0.35 MPa. Compared with the original sample, the indigo and indirubin contents in purified Indigo Naturalis were 1.6 and 3 times higher, the total ash content decreased from 86 to 70%, the pH value decreased from 12.18 to 9.71, and the leachate doubled. Animal experiments suggested the significantly reduced irritation (p < 0.01) and enhanced antioxidative stress activity (p < 0.01) of Indigo Naturalis after foam separation. Therefore, the foam-separation equipment developed in this study enabled the refinement of active ingredients in Indigo Naturalis, which greatly improved the production efficiency and quality.

Combination Therapy with Indigo and Indirubin for Ulcerative Colitis via Reinforcing Intestinal Barrier Function.

Indigo and indirubin, the active molecules of traditional Chinese medicine indigo naturalis, exert therapeutic activity for ulcerative colitis (UC). Indigo and indirubin are isomers and have distinctive profiles in anti-inflammation, immune regulation, intestinal microbiota regulation, oxidative stress regulation, and intestinal mucosal repair for UC treatment. Thus, exploring its combined administration's integrated advantages for UC is critical. This study is aimed at clarifying the effect and mechanisms of the combined administration of indigo and indirubin on colitis mouse models. The results showed that all the treatment groups could improve the disease symptoms, and the combined administration showed the best effect. Additionally, compared with indigo and indirubin alone, the combination group could significantly reinforce intestinal barrier function by increasing the expression of E-cadherin, occludin, ZO-1, and MUC2 and improving intestinal permeability. The treatment groups significantly improved the expression of cytokines, including TNF-α, IFN-γ, IL-12, IL-23, and IL-17A, and indirubin presented the most potent anti-inflammatory effect. Furthermore, all the treatment groups reduced the infiltration of the immune cells in intestinal lamina propria and the production of ROS/RNS. Notably, indigo exhibited a more substantial capacity to regulate natural killer (NK) cells, ILC3, neutrophils, and dendritic cells, followed by the combination group and indirubin alone. Finally, all the treatment groups modulated intestinal microbiota composition, increased the proportion of beneficial microbiota, and decreased the proportion of microbiota. Our results indicated that indigo and indirubin synergistically reinforced the intestinal barrier function, which may be associated with integrating the indirubin anti-inflammatory and intestinal microbiota regulating strength and indigo immune and ROS/RNS regulation advantage.

[Microbial community structure and transformation of indoles in soaking and fermentation of Indigo Naturalis].

The soaking and fermentation of Baphicacanthus cusia( Nees),the important intermediate link of Indigo Naturalis processing,facilitates the synthesis of indigo and indirubin precursors and the dissolution of endogenous enzymes and other effective components,while the role of microorganisms in the fermentation is ignored. The present study investigated the changes of microbial community structure in Indigo Naturalis processing based on 16 S amplicon sequencing and bioinformatics. Meanwhile,the contents of indigo,indirubin,isatin,tryptanthrin,indole glycoside,etc. were determined to explore the correlation between the microorganisms and the alterations of the main components. As demonstrated by the results,the microbial diversity decreased gradually with the fermentation,which bottomed out after the addition of lime. Proteobacteria,Bacteroidetes,and Firmicutes were the main dominant communities in the fermentation. The relative abundance of Proteobacteria declined gradually with the prolongation of fermentation time,and to the lowest level after the addition of lime. The relative abundance of Firmicutes increased,and that of Bacteroidetes decreased first and then increased. The contents of effective substances in Indigo Naturalis also showed different variation tendencies. As fermentation went on,indole glycoside decreased gradually; indigo first increased and then decreased; indirubin and isatin first decreased and then increased; tryptanthrin gradually increased. Those changes were presumedly related to the roles of microorganisms in the synthesis of different components. This study preliminarily clarified the important role of microorganisms in the soaking and fermentation and provided a scientific basis for the control of Indigo Naturalis processing and the preparation of high-quality Indigo Naturalis.

[Antipyretic activity and potential mechanism of Indigo Naturalis on 2,4-dinitrophenol-induced fever rat model].

As an effective antipyretic medicine,Indigo Naturalis has a long history of application in the field of Chinese medicine.The content of organics,mainly indigo and indirubin,is about 10%. However,the active ingredients and mechanism of its antipyretic effect have not yet been fully elucidated. In view of this,they were investigated in this study with the rectal temperature change as an indicator and 2,4-dinitrophenol-induced fever rats as subjects. The content of PGE2 and c AMP in the hypothalamus and the serum levels of TNF-α,IL-1ß and IL-6 were determined by ELISA. Moreover,the plasma samples of fever rats were analyzed by metabonomics in combination with UPLC-Q-TOF-MS for the exploration of potential biomarkers and the discussion on the antipyretic mechanism of Indigo Naturalis and its active ingredients. The results showed that the rising trend of rectal temperature in rats was suppressed 0. 5 h after the treatment with Indigo Naturalis,organic matter,indigo or indirubin as compared with the rats of model group( P < 0. 05),among which Indigo Naturalis and organic matter had better antipyretic effect. ELISA results showed that organic matter and indigo can inhibit the expression of PGE2 and c AMP( P<0. 01),while Indigo Naturalis and organic matter were effective in curbing the increase in TNF-α( P<0. 05). A total of 21 endogenous metabolites were identified from the plasma samples of the Indigo Naturalis,organic matter,indigo and indirubin groups,which were mainly involved in glycerophospholipid metabolism.

Rhizoma coptidis as a Potential Treatment Agent for Type 2 Diabetes Mellitus and the Underlying Mechanisms: A Review.

Diabetes mellitus, especially type 2 diabetes mellitus (T2DM), has become a significant public health burden. Rhizoma coptidis (RC), known as Huang Lian, is widely used for treating diabetes in China. The bioactive compounds of RC, especially alkaloids, have the potential to suppress T2DM-induced lesions, including diabetic vascular dysfunction, diabetic heart disease, diabetic hyperlipidemia, diabetic nephropathy, diabetic encephalopathy, diabetic osteopathy, diabetic enteropathy, and diabetic retinopathy. This review summarizes the effects of RC and its bioactive compounds on T2DM and T2DM complications. Less research has been conducted on non-alkaloid fractions of RC, which may exert synergistic action with alkaloids. Moreover, we summarized the pharmacokinetic properties and structure-activity relationships of RC on T2DM with reference to extant literature and showed clearly that RC has potential therapeutic effect on T2DM.

Coptidis Rhizoma: a comprehensive review of its traditional uses, botany, phytochemistry, pharmacology and toxicology.

CONTEXT: Coptidis rhizome (CR), also known as Huanglian in Chinese, is the rhizome of Coptis chinensis Franch., C. deltoidea C.Y. Cheng et Hsiao, or C. teeta Wall (Ranunculaceae). It has been widely used to treat bacillary dysentery, diabetes, pertussis, sore throat, aphtha, and eczema in China. OBJECTIVES: The present paper reviews the latest advances of CR, focusing on the botany, phytochemistry, traditional usages, pharmacokinetics, pharmacology and toxicology of CR and its future perspectives. METHODS: Studies from 1985 to 2018 were reviewed from books; PhD. and MSc. dissertations; the state and local drug standards; PubMed; CNKI; Scopus; the Web of Science; and Google Scholar using the keywords Coptis, Coptidis Rhizoma, Huanglian, and goldthread. RESULTS: Currently, 128 chemical constituents have been isolated and identified from CR. Alkaloids are the characteristic components, together with organic acids, coumarins, phenylpropanoids and quinones. The extracts/compounds isolated from CR cover a wide pharmacological spectrum, including antibacterial, antivirus, antifungal, antidiabetic, anticancer and cardioprotective effects. Berberine is the most important active constituent and the primary toxic component of CR. CONCLUSIONS: As an important herbal medicine in Chinese medicine, CR has the potential to treat various diseases. However, further research should be undertaken to investigate the clinical effects, toxic constituents, target organs and pharmacokinetics, and to establish criteria for quality control, for CR and its related medications. In addition, the active constituents, other than alkaloids, in both raw and processed products of CR should be investigated.

[Effects of plant growth regulators and nitrogen on germination of Epimedium pseudowushanense after its stems and leaves reaped].

In order to explore appropriate measures to promote germination after the harvest of Epimedium pseudowushanense, 6-BA, urea, ammonium bicarbonate and GA3 were chosen to spray on the root and rhizomes, and then the biological indicators such as branches, leaf length, leaf width, plant height and so on, were measured in different periods, and the contents of epimedin A, epimedin B, epimedin C and icariin in the dry leaves were detected by HPLC. Results showed that 6-BA 90 mg·L⁻¹(B1), 6-BA 60 mg·L⁻¹(B2),6-BA 30 mg·L⁻¹+urea 300 mg·L⁻¹ (C1), 6-BA 60 mg·L⁻¹+urea 300 mg·L⁻¹(C2),6-BA 60 mg·L⁻¹+ ammonium bicarbonate 300 mg·L⁻¹(C4) significantly increased bud germination in the early period, and the plants quickly set up new system of photosynthesis, the branches in a month of which were higher than the control group respectively by 165.9%, 115.76%, 103.86%, 104.50%, 81.67%.However the branches developed the next year and the dry weight of leaves per plant in group B1 and B2 were much lower than that in control group. The groups that use 6-BA and nitrogen at the same time reaped a good yield of leaves even though the treatment had no significant influence on the branches developed the next year. The dry weight of leaves of C1, C2, C4 treatments were 36.80%, 32.84%, 45.97% more than the control group respectively. Therefore, C1, C2 and C4 treatments are the more appropriate to promote recovery after harvest. Furthermore, different groups, except 10 mg·L⁻¹ 6-BA treatment significantly reduced the content of epimedin C, other groups didn't have any significant effect on the contents of such flavonoids.

[Genetic relationship and population genetic diversity of Epimedium pubescens and its related species based on ISSR and non-glandular hair characteristic].

In this study, 128 individuals form 14 Epimedium pubescens populations and 1 E. stellulatum population were analyzed by ISSR marker. The data were calculated by POPGENE software and clustered by UPGMA method. Optical microscope was used to observe the main types of the non-glandular hairs and their characteristics in each population. It is found that the following conclusions: Non-glandular hairs can be divided into five morphological categories, long straight pubescent, curly pubescent, appressed curly pubescent, pseudo short appressed hairs and long appressed. Eight primers were screening and a total of 94 bands were detected in ISSR, among which 90 were polymorphic bands. Based on the results of ISSR cluster analysis, 15 populations were divided into 3 clades. E. stellulatum populations should be incorporated into the E. pubescens or as avariety under E. pubescens not be independent and as it has no separate phylogenetic branch for a cluster. The genetic relationship among the populations of E. pubescens was closely related with its geographical distribution and non-glandular hair features. But there were also some inconsistency, which provided a good hint for the further study on the interspecific relationship and natural speciation manner of Epimedium species. Population diversity analysis showed Nm=0.354 4, Nei's=0.585 2. It was showed that E. pubescens has high genetic diversity among populations, for which the main reason was probably the high inbreeding rate and the small range of seed dispersal.

[Effects of nitrogen on yields and flavonoids contents of Epimedium pseudowushanense in hydroponic conditions].

Nitrogen nutrition divided into concentrations and forms for Epimedium pseudowushanense was studied under hydroponic conditions. There were 7 nitrogen concentrations treatments（N0-N6）which nitrogen concentration was 0, 2.5, 5.0, 7.5, 10.0, 13.0 mmol•L⁻¹, respectively, and 5 nitrogen forms and proportions treatments (T1-T5) which NO3-N/NH4-N was 5∶0, 4∶1, 2.5∶2.5, 1∶4, 0∶5 respectively, the total nitrogen applied was 5 mmol•L⁻¹. The results showed that along with the increase of nitrogen concentration, biomass (dry weight) and leaf dry weight increased, which in N5 treatment was 29%, 23% higher than those in N0 and 36%, 23% higher than those in N4 respectively, but not significantly different from N6 treatment. Roots and stem dry weights did not change significantly. Pn and the contents of photosynthetic pigments were higher in N3-N5 treatments. Epimedin A, B, C, icariin and total flavonoids contents were the lowest in N3 treatment, suddenly increase in N4 treatment and only a little change between N4-N6 treatments. N5 treatment with the highest epimedin C content was 131% higher than N3 treatment with the lowest epimedin C content (P<0.05). While in the nitrogen forms and proportions treatments, biomass, leaf and stem dry weights were the highest in T3 treatment and lower in T1 and T5 treatments, without significance between T1 and T5 treatments. These indicators in T3 treatment were more than 30% higher than those in T1 treatment. The impact of nitrogen concentrations was greater than nitrogen forms on flavonoids. epmedin C content was the highest in T1 treatment, only 5%-8% higher than that in T2-T5 treatments (no significance between 4 treatments). Epimedin A, B, icariin and total flavonoids contents were higher in T1 treatment, the lowest in T2 treatment, those in T1 treatment was 41%, 62% and 27% higher than T2 treatment respectively (P<0.05). In a conclusion, E. pseudowushanense is a high nitrogen tolerant plant, 10.0 mmol•L⁻¹ was the boundary point between growing general to well and the optimal nitrogen concentration for it was 10.0-13.0 mmol•L⁻¹. Besides, it prefers to grow in NO-3 plus NH+4 and the optimal proportion was 2.5∶2.5.

In vitro and in vivo chemo-phototherapy of magnetic TiO2 drug delivery system formed by pH-sensitive coordination bond.

To achieve tumor-specific delivery of doxorubicin, TiO2@Fe3O4/PEI/delivery of doxorubicin conjugates were designed and synthesized. Fe3O4 could act as magnetically responsive carriers and enhance the visible light photodynamic activities of TiO2 Delivery of doxorubicin was conjugated via coordination bond. The drug release rate at pH 5.2 was much faster than that at pH 7.4, due to pH-sensitive coordination bond. Besides, TiO2@Fe3O4/PEI/delivery of doxorubicin showed high antitumor efficacy combining with phototherapy, good bio-safety, higher cellular uptake with an external magnetic field, and less toxicity in vitro and in vivo. These results suggested that TiO2@Fe3O4/PEI/delivery of doxorubicin may be promising for high tumor treatment efficacy with minimal side effects in future.

[Research on photosynthetic and eco-physiological characteristics of Epimedium pseudowushannense in different growing age].

Epimedium is a widely used medicinal plants. Due to excessive use of wild resources and resource constraints, artificial cultivation is the only way to achieve sustainable use of resources. E. pseudowushannense is intended to achieve the first cultivated species, but plant production technology is not yet fully mature.The diurnal variation of photosynthetic and eco-physiological characteristics of E. pseudowushannense in different growing age was studied by the CI-340 Portable Photosynthesis System under natural conditions. The net photosynthetic rate (Pn), stomatal conductance (C), intercellular CO2 concentration (Ci), photosynthetically active radiation (PAR), relative humidity (RH), air temperature (Ta), apparent mesophyll conductance (ALMC), transpiration rate (E), leaf temperature (Tl), water use efficiency (WUE) and other photosynthetic physiological factors and environmental factors were measured. The obtained data was analyzed by correlation analysis, path analysis and stepwise multiple-regression analysis to explore the relationship between net photosynthetic rate and physiological and ecological factors. The results show that： ①The daily variation of the Pn, E, ALMC, C of E. pseudowushannense in different growing age took on"two peak type", the Pn was with an obvious midday depression phenomenon at noon.②The Pn, ALMC and C of annuals' were significantly higher than biennial or three-year; but the Ci was basically a three-year > biennial> annual.③The correlation analysis, path analysis and stepwise regression analysis showed that ALMC, Ci, Ta might played important role in the Pn and ranked as ALMC>Ci>Ta.

[Distribution and influencing factors of magnoflorine in Epimedium].

Eighty samples of Epimedium from 29 species and were determined in this study. The content of magnoflorine in leaves range between 0. 003% and 2. 603%. The results showed that the content of magnoflorine was quite stable within species except E. wushanense, E. acuminatum, E. hunanense. Genetic factors might be the main influencing ones. The contents of different parts and different collecting time of the medicinal materials were variable.

The alleviating pain effect of aqueous extract from tong-xie-yao-fang, on experimental visceral hypersensitivity and its mechanism.

Tong-Xie-Yao-Fang (TXYF) is a prescription in traditional chinese medicine (TCM), used for relieving abdominal pain associated with irritable bowel syndrome. The aim of the present study was to investigate the effects and mechanism of TXYF on experimental visceral hypersensitivity (VH) models. TXYF affected the abdominal withdrawal reflex produced by colonic distention in maternal separation-induced visceral hypersensitivity rats, in a dosage-dependent manner. TXYF significantly decreased serotonin (5-HT) levels in serum and corticotrophin releasing factor (CRF) concentrations in the brain. Moreover, it was found that VH alleviation by TXYF was dependent on the substance P (SP) expression in the colon mucosa. These results suggest that TXYF attenuates behavioral hyperalgesia by regulating substance associated with the brain-gut axis, including decreasing the expression of 5-HT and SP in the periphery and that of CRF in the center.