Establishment of XRD fourier fingerprint identification method of realgar decoction pieces and its anti-tumor activity in tumor-in-situ transplanted mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Realgar, a traditional mineral Chinese medicine, has been used in China for more than 2000 years. It has been recorded in many ancient and modern works that it has anti-cancer and anti-tumor effects. Of course, colon cancer is also within the scope of its treatment. Realgar needs to be processed into realgar decoction pieces by water grinding before being used for medicine. To ensure the consistency of efficacy and quality of realgar decoction pieces, modern methods need to be used for further quality control. AIM OF THE STUDY: The research of traditional mineral Chinese medicine is relatively difficult, and the related research is less. The purpose of this study is to control the quality of realgar decoction pieces by modern analytical technology and analyze its components. On this basis, its anti-colon cancer activity was discussed. MATERIALS AND METHODS: Several batches of realgar decoction pieces were analyzed by XRD, and the components of realgar decoction pieces were obtained. The quality control fingerprints of realgar decoction pieces were established by processing XRD spectra and similarity evaluation. Then, the effects of realgar decoction pieces on apoptosis of CT26 and HTC-116 cells were observed in vitro by Hoechst 33258 staining, flow cytometry, measurement of mitochondrial membrane potential and Western blot; In vivo, the mouse model of tumor-in-situ transplantation of colon cancer was established, and the related indexes were observed. RESULT: The explorations showed that the XRD Fourier fingerprints of realgar decoction pieces samples that had the same phase revealed 10 common peaks, respectively. The similarity evaluation of the established XRD Fourier fingerprint was greater than 0.900. We also demonstrated that realgar decoction pieces can promote apoptosis and inhibit tumor growth in colon cancer cells, its activating effect on p53 protein, and its safety when used within reasonable limits. CONCLUSION: The quality control of realgar decoction pieces by XRD is scientific and has the inhibitory effect on colon cancer, which has the development potential.

Recent advance in phytonanomedicine and mineral nanomedicine delivery system of the treatment for acute myeloid leukemia.

Acute myeloid leukemia (AML) is an invasive hematopoietic malignancy caused by excessive proliferation of myeloblasts. Classical chemotherapies and cell transplantation therapies have remarkable efficacy in AML treatment; however, 30-40% of patients relapsed or had refractory disease. The resistance of AML is closely related to its inherent cytogenetics or various gene mutations. Recently, phytonanomedicine are found to be effective against resistant AML cells and have become a research focus for nanotechnology development to improve their properties, such as increasing solubility, improving absorption, enhancing bioavailability, and maintaining sustained release and targeting. These novel phytonanomedicine and mineral nanomedicine, including nanocrystals, nanoemulsion, nanoparticles, nanoliposome, and nanomicelles, offer many advantages, such as flexible dosages or forms, multiple routes of administration, and curative effects. Therefore, we reviewed the application and progress of phytomedicine in AML treatment and discussed the limitations and future prospects. This review may provide a solid reference to guide future research on AML treatment.

Traditional mineral medicine realgar and Realgar-Indigo naturalis formula potentially exerted therapeutic effects by altering the gut microbiota.

Introduction: Realgar has a long history ofuse in traditional medicines. However, the mechanism through which Realgar or Realgar-Indigo naturalis formula (RIF) exert therapeutic effects is only partially understood. Methods: In this study, 60 feces and 60 ileum samples from rats administered with realgar or RIF were collected to examine the gut microbiota. Results: The results showed that realgar and RIF influenced different microbiota in both feces and ileum. Compared with realgar, RIF at low dosage (0.1701 g/3 ml) significantly increased the microbiota diversity. LEfSe and random forest analyses showed that the bacterium Bacteroidales was significantly altered after RIF administration, and it was predicted that these microorganisms contribute to the inorganic arsenic metabolic process. Discussion: Our results suggest that realgar and RIF may exert their therapeutic effects through influencing microbiota. The low dose of RIF had greater effects on increasing the diversity of microbiota, and Bacteroidales in feces might participate in the inorganic arsenic metabolic process to exert therapeutic effects for realgar.

Analysis of Polarizing Microscopic Characteristics and X-ray Diffraction Fingerprint of Mineral Medicine Maifanitum / 中国实验方剂学杂志

ObjectiveTo analyze the polarized light microscopic characteristics， the composition of physical phases and their relative contents of Maifanitum from different origins， and to establish the Fourier characteristic fingerprint of Maifanitum powder crystals by X-ray diffraction（XRD）. MethodA total of 26 batches of Maifanitum samples were selected， and the microscopic characteristics of the sample powders and grinding flakes were observed by polarized light microscopy under single polarized light and orthogonal polarized light， and the main phase compositions and their relative contents were analyzed by powder crystal XRD technique， and the XRD Fourier characteristic fingerprint of Maifanitum was established. The incident light source of XRD was Cu target Kβ radiation， the light tube voltage and light tube current were 40 kV and 40 mA， respectively， the divergence slit was 1°， the scattering slit was 1°， the receiving slit was 0.2 mm， the scanning speed was 5°·min-1 with continuous scanning and scanning range of 5-90°（2θ）， and the step length was 0.02°. ResultThe polarized light micrographs of powders and grinding flakes of Maifanitum were obtained， and the main phases were plagioclase， potassium feldspar and quartz， and a few samples also contained illite， pyrite， iron dolomite， calcite， iron amphibole and chlorite， etc. The relative total content of feldspar phases was 61.9%-82.4%， and the relative content of quartz was 12.6%-33.6%. The XRD Fourier fingerprint analysis method of Maifanitum with 13 common peaks as the characteristic fingerprint information was established， and the similarity calculated by the mean correlation coefficient method was 0.920 9-0.997 7， the similarity calculated by the mean angle cosine method was 0.940 5-0.998 4， the similarity calculated by the median correlation coefficient method was 0.921 1-0.997 5， and the similarity calculated by the median angle cosine method was 0.947 5-0.998 2. ConclusionThe polarized light microscopic identification characteristics of Maifanitum are mainly plagioclase， quartz and potassium feldspar， and the technique of powder crystal XRD Fourier fingerprint analysis can be used for the identification of Maifanitum.

Jinhua Qinggan granules for non-hospitalized COVID-19 patients: A double-blind, placebo-controlled, and randomized controlled trial.

Background: Key findings from the World Health Organization Expert Meeting on Evaluation of Traditional Chinese Medicine (TCM) in treating coronavirus disease 2019 (COVID-19) reported that TCMs are beneficial, particularly for mild-to-moderate cases. The efficacy of Jinhua Qinggan granules (JHQG) in COVID-19 patients with mild symptoms has yet to be clearly defined. Methods: We conducted a phase 2/3, double-blind, randomized, placebo-controlled trial to evaluate the efficacy and safety of treatment with JHQG in mild, non-hospitalized, laboratory-confirmed COVID-19 patients. Participants were randomly assigned to receive 5 g/sacket of JHQG or placebo granules orally thrice daily for 10 days. The primary outcomes were the improvement in clinical symptoms and a proportion tested negative on viral polymerase chain reaction (PCR) after treatment. Secondary outcomes were the time to recover from clinical symptoms and changes in white blood cells (WBC) and acute phase reactants (C-reactive protein (CRP) and ferritin) on the 10th day after treatment initiation. Results: A total of 300 patients were randomly assigned to receive JHQG (150 patients) and placebo (150 patients). Baseline characteristics were similar in the two groups. In the modified intention-to-treat analysis, JHQG showed greater clinical efficacy (82.67%) on the 10th day of the trial compared with the placebo group (10.74%; rate difference: 71.93%; 95% CI 64.09-79.76). The proportion of patients with a negative PCR after treatment was comparable (rate difference: -4.67%; 95% CI -15.76 to 6.42). In contrast, all changes in WBC, ferritin, and CRP levels showed a statistically significant decline in JHQG (P ≤ 0.044) after treatment, but not the latter in placebo (P = 0.077). The median time to recovery of COVID-19-related symptoms including cough, sputum, sore throat, dyspnea, headache, nasal obstruction, fatigue, and myalgia was shorter in the JHQG group compared to the placebo group (P < 0.001 for all). Three patients experienced mild-to-moderate adverse events (AEs) duringthe treatment period in the JHQG group. Findings were similar between the modified intention-to-treat and the per-protocol analysis that included only patients who reported 100% adherence to the assigned regimen. Conclusion: Based on the time to recover from the COVID-19-related symptoms and AEs, it is concluded that JHQG is a safe and effective TCM for symptomatic relief of patients with mild COVID-19. A symptomatic improvement in the JHQG group patients was observed and JHQG use would have important public health implications in such patients. Clinical Trial Registration: The Trial was prospectively registered on www.clinicaltrials.gov with registration number: NCT04723524.

Chinese Medicine, Succinum, Ameliorates Cognitive Impairment of Carotid Artery Ligation Rats and Inhibits Apoptosis of HT22 Hippocampal Cells via Regulation of the GSK3ß/ß-Catenin Pathway.

Succinum is an organic mineral formed from the resin of ancient coniferous and leguminous plants, which is applied for tranquilizing mood, promoting blood circulation, and removing blood stasis in Chinese medicine. For quite a long time, the modern research of succinum mainly focuses on the study of physical and chemical properties and authenticity identification while few reports on its medicinal mechanism. In current study, we evaluated different solvent extracts of succinum on carotid artery ligation rats mimicking vascular dementia. It was found that ethyl acetate extracts of succinum significantly improved the learning and memory abilities of model rats and inhibited neuronal apoptosis in the hippocampus. On a mice hippocampal neuronal cell line (HT22), ethyl acetate extracts of succinum also exerted better action trend in inhibiting cell apoptosis induced by oxygen glucose deprivation (OGD). By using XAV-939 on both in vivo and in vitro studies, it was found that ethyl acetate extracts of succinum might exert these functions by regulating the GSK3ß/ß-catenin pathway. These studies revealed the neuronal function of succinum, which explained the traditional effects of succinum and provided more modern scientific basis for its clinical application.

Research Progress of Gypsum Ustum and Application of Calcium-containing Hemostatic Materials / 中国实验方剂学杂志

Mineral medicine is an indispensable part of traditional Chinese medicine and has a long history of application. Among them， mineral-based hemostatics have been widely applied for the treatment of various hemorrhagic diseases with extensive clinical experience and established efficacy. Gypsum Fibrosum （GF）， a commonly used mineral medicine in clinical， can clear away heat， and relieve anxiety and thirst. Gypsum Ustum （GU） is the processed product of GF after calcining at high temperature. It is mainly composed of anhydrous calcium sulfate （CaSO4） with the functions of moisturizing， promoting muscle growth， astringent sores and hemostasis. GU is often used externally to treat ulcer， itching， eczema， water and fire scalds， trauma bleeding， etc. Studies on the mechanism of hemostasis have shown that Ca2+ （coagulation factor Ⅳ） is involved in many key processes of the internal and external coagulation cascades and can prevent bleeding by regulating platelet activation and aggregation， and promoting the production of insoluble fibrin and the ultimate formation of a blood clot. GF and GU both contain Ca2+ which provide an important material basis of hemostatic effect for both compounds， but GU has a significant hemostatic effect， while GF has no hemostatic effect. After processing， the taste and efficacy of the GF have been obviously changed which reflects the characteristics of processing， but the processing mechanism of GU has not been fully clarified. Therefore， based on studies of GF before and after calcining， this paper focused on these aspects including calcining process， crystal form comparison， element content， efficacy comparison， and summarized various aspects of Ca2+ involved in hemostasis. In addition， the hemostatic properties of other calcium-containing mineral medicines and new calcium-containing hemostatic materials such as calcium alginate， mesoporous calcium silicate and nanogel hemostatic materials were also discussed. The paper aimed to provide a reference for elucidating processing mechanism and clinical dialectical use of GU， also to promote development of new calcium-containing hemostatic materials.

Distribution of medicinal mineral resources in Jiangsu Province and their utilization status and prospects / 中草药

Jiangsu Province is rich in medicinal mineral resources because of its complex geological conditions. With the aid of the fourth national survey of traditional Chinese medicine resources and other projects, literature studies have found nearly 90 species of medicinal mineral resources in Jiangsu Province. Through the survey of mineral medicinal resources in this study, there were nearly 70 kinds of medicinal mineral resources in Jiangsu Province, and their utilization status was analyzed. The investigation is of great social and economic significance for carrying forward the traditional mineral medicine of the motherland, fully tapping the medicinal value of Jiangsu Province’s medicinal mineral resources and developing geological economy. At the same time, it provides reference for the formulation, promotion and production layout of Jiangsu Province’s regional mineral drug resources industrialization development plan.

Bioleaching of realgar nanoparticles using the extremophilic bacterium Acidithiobacillus ferrooxidans DLC-5

BACKGROUND: This paper presents micro- and nano-fabrication techniques for leachable realgar using the extremophilic bacterium Acidithiobacillus ferrooxidans (A. ferrooxidans) DLC-5. RESULTS: Realgar nanoparticles of size ranging from 120 nm to 200 nm were successfully prepared using the highenergy ball mill instrument. A. ferrooxidans DLC-5 was then used to bioleach the particles. The arsenic concentration in the bioleaching system was found to be increased significantly when compared with that in the sterile control. Furthermore, in the comparison with the bioleaching of raw realgar, nanoparticles could achieve the same effect with only one fifth of the consumption. CONCLUSION: Emphasis was placed on improving the dissolvability of arsenic because of the great potential of leachable realgar drug delivery in both laboratory and industrial settings

Determination of zinc oxide content of mineral medicine calamine using near-infrared spectroscopy based on MIV and BP-ANN algorithm.

Near-infrared (NIR) spectroscopy has been widely used in the analysis fields of traditional Chinese medicine. It has the advantages of fast analysis, no damage to samples and no pollution. In this research, a fast quantitative model for zinc oxide (ZnO) content in mineral medicine calamine was explored based on NIR spectroscopy. NIR spectra of 57 batches of calamine samples were collected and the first derivative (FD) method was adopted for conducting spectral pretreatment. The content of ZnO in calamine sample was determined using ethylenediaminetetraacetic acid (EDTA) titration and taken as reference value of NIR spectroscopy. 57 batches of calamine samples were categorized into calibration and prediction set using the Kennard-Stone (K-S) algorithm. Firstly, in the calibration set, to calculate the correlation coefficient (r) between the absorbance value and the ZnO content of corresponding samples at each wave number. Next, according to the square correlation coefficient (r2) value to obtain the top 50 wave numbers to compose the characteristic spectral bands (4081.8-4096.3, 4188.9-4274.7, 4335.4, 4763.6,4794.4-4802.1, 4809.9, 4817.6-4875.4cm-1), which were used to establish the quantitative model of ZnO content using back propagation artificial neural network (BP-ANN) algorithm. Then, the 50 wave numbers were operated by the mean impact value (MIV) algorithm to choose wave numbers whose absolute value of MIV greater than or equal to 25, to obtain the optimal characteristic spectral bands (4875.4-4836.9, 4223.6-4080.9cm-1). And then, both internal cross and external validation were used to screen the number of hidden layer nodes of BP-ANN. Finally, the number 4 of hidden layer nodes was chosen as the best. At last, the BP-ANN model was found to enjoy a high accuracy and strong forecasting capacity for analyzing ZnO content in calamine samples ranging within 42.05-69.98%, with relative mean square error of cross validation (RMSECV) of 1.66% and coefficient of determination (R2) of 95.75% in internal cross and relative mean square error of prediction (RMSEP) of 1.98%, R2 of 97.94% and ratio of performance to deviation (RPD) of 6.11 in external validation.

[Modern research and application of Longgu].

Longgu is the fossil of ancient mammals which was used as a common kind of mineral medicine. Longgu is always used to treat neurological diseases. Currently, the quality standard of Longgu is incomplete. Moreover, because of the non-renewable nature of the resource and the increase of national protection of fossils, the clinical application of Longgu is facing a series of problems. As the discovery of the ingredient and the development of forging technology researchers launched to search the substitutes of Longgu. The article summarizes the usage and the study of Longgu, in order that we can discuss the modern usage and substitutability of Longgu.

Determination of Magnesium Oxide Content in Mineral Medicine Talcum Using Near-Infrared Spectroscopy Integrated with Support Vector Machine.

In this research paper, a fast, quantitative, analytical model for magnesium oxide (MgO) content in medicinal mineral talcum was explored based on near-infrared (NIR) spectroscopy. MgO content in each sample was determined by ethylenediaminetetraacetic acid (EDTA) titration and taken as reference value of NIR spectroscopy, and then a variety of processing methods of spectra data were compared to establish a good NIR spectroscopy model. To start, 50 batches of talcum samples were categorized into training set and test set using the Kennard-Stone (K-S) algorithm. In a partial least squares regression (PLSR) model, both leave-one-out cross-validation (LOOCV) and training set validation (TSV) were used to screen spectrum preprocessing methods from multiplicative scatter correction (MSC), and finally the standard normal variate transformation (SNV) was chosen as the optimal pretreatment method. The modeling spectrum bands and ranks were optimized using PLSR method, and the characteristic spectrum ranges were determined as 11995-10664, 7991-6661, and 4326-3999 cm-1, with four optimal ranks. In the support vector machine (SVM) model, the radical basis function (RBF) kernel function was used. Moreover, the full spectrum data of samples pretreated with SNV, the characteristic spectrum data screened using synergy interval partial least squares (SiPLS), and the scoring data of the first four ranks obtained by a partial least squares (PLS) dimension reduction of characteristic spectrum were taken as input variables of SVM, and the MgO content reference values of various sample were taken as output values. In addition, the SVM model internal parameters were optimized using the grid optimization method (GRID), particle swarm optimization (PSO), and genetic algorithm (GA) so that the optimal C and g-values were determined and the validation model was established. By comprehensively comparing the validation effects of different models, it can be concluded that the scoring data of the first four ranks obtained by PLS dimension reduction of characteristic spectrum were taken as input variables of SVM, and the PLS-SVM regression model established using GRID was the optimal NIR spectroscopy quantitative model of talc. This PLS-SVM regression model (rank = 4) measured that the MgO content of talcum was in the range of 17.42-33.22%, with root mean square error of cross validation (RMSECV) of 2.2127%, root mean square error of calibration (RMSEC) of 0.6057%, and root mean square error of prediction (RMSEP) of 1.2901%. This model showed high accuracy and strong prediction capacity, which can be used for rapid prediction of MgO content in talcum.

A Rapid Identification Method for Calamine Using Near-Infrared Spectroscopy Based on Multi-Reference Correlation Coefficient Method and Back Propagation Artificial Neural Network.

As a mineral, the traditional Chinese medicine calamine has a similar shape to many other minerals. Investigations of commercially available calamine samples have shown that there are many fake and inferior calamine goods sold on the market. The conventional identification method for calamine is complicated, therefore as a result of the large scale of calamine samples, a rapid identification method is needed. To establish a qualitative model using near-infrared (NIR) spectroscopy for rapid identification of various calamine samples, large quantities of calamine samples including crude products, counterfeits and processed products were collected and correctly identified using the physicochemical and powder X-ray diffraction method. The NIR spectroscopy method was used to analyze these samples by combining the multi-reference correlation coefficient (MRCC) method and the error back propagation artificial neural network algorithm (BP-ANN), so as to realize the qualitative identification of calamine samples. The accuracy rate of the model based on NIR and MRCC methods was 85%; in addition, the model, which took comprehensive multiple factors into consideration, can be used to identify crude calamine products, its counterfeits and processed products. Furthermore, by in-putting the correlation coefficients of multiple references as the spectral feature data of samples into BP-ANN, a BP-ANN model of qualitative identification was established, of which the accuracy rate was increased to 95%. The MRCC method can be used as a NIR-based method in the process of BP-ANN modeling.

Modern research and application of Longgu / 中国中药杂志

Longgu is the fossil of ancient mammals which was used as a common kind of mineral medicine. Longgu is always used to treat neurological diseases. Currently, the quality standard of Longgu is incomplete. Moreover, because of the non-renewable nature of the resource and the increase of national protection of fossils, the clinical application of Longgu is facing a series of problems. As the discovery of the ingredient and the development of forging technology researchers launched to search the substitutes of Longgu. The article summarizes the usage and the study of Longgu, in order that we can discuss the modern usage and substitutability of Longgu.

Monitoring As and Hg variation in An-Gong-Niu-Huang Wan (AGNH) intermediates in a pilot scale blending process using laser-induced breakdown spectroscopy.

Laser-induced breakdown spectroscopy (LIBS) was used to assess the cinnabar and realgar blending of An-Gong-Niu-Huang Wan (AGNH) in a pilot-scale experiment, including the blending end-point. The blending variability of two mineral medicines, cinnabar and realgar, were measured by signal relative intensity changing rate (RICR) and moving window standard deviation (MWSD) based on LIBS. Meanwhile, relative concentration changing rate (RCCR) was obtained based on the reference method involving inductively coupled plasma optical emission spectrometry (ICP-OES). The LIBS result was consistent with ICP-OES at blending end-point determinations of both mineral medicines. Unlike the ICP-OES method, LIBS does not have an elaborate digestion procedure. LIBS is a promising and rapid technique to understand the blending process of Chinese Materia Medica (CMM) containing cinnabar and realgar. These results demonstrate the potential of LIBS in monitoring CMM pharmaceutical production.

Current situation analysis and countermeasures on contamination of heavy metal in traditional Chinese medicinal materials in China / 中草药

Recent years, with the rapid development of industrialization in China, the contamination of heavy metal was worsening, and traditional Chinese medicinal materials (TCMM) were also suffered. The data were collected in the literatures in the past five years, and Green Trade Standard of Importing & Exporting Medicinal Plants & Preparations was used as the standard to evaluate the pollution conditions of lead (Pb), cadmium (Cd), mercury (Hg), arsenic (As), and copper (Cu). The five heavy metals exceeded the limits of the standard and the percentages were 9.66%, 26.35%, 13.0%, 9.32%, and 16.09%, respectively. Moreover, statistics on the residues of the heavy metals in different categories of TCMM from various origins were also carried out in this paper. The results showed that the contamination of heavy metals in TCMM differently depended on the origins and categories. Studies on the rapid detection methods and the limit standards of heavy metal should be established immediately. At the same time, the regulations should be formulated timely and supervision also should be intensified. To solve the problems of excessive amounts of heavy metals in TCMM, necessary measures should be taken to control the contamination of TCMM.

Study on Identification Method of 7 Kinds of Sulfates Mineral Medicines by Near-infrared Spectroscopy / 世界科学技术-中医药现代化

To establish seven kinds of minerals containing sulfate kind of near infrared spectral identification method of traditional Chinese medicine (TCM). 7 species of mineral medicine containing sulfate after crushing sieving, measure all the samples in 12 000-4 000 cm-1 section within the scope of the near infrared spectrum, spectrum signal by different pretreatment methods, after the screening of the different characteristics of the spectrum to extract the effective information, using cluster analysis method for qualitative identification. In 8 600-8 100 cm-1, 5 843-4 245 cm-1, 7 096-6 337 cm -1 section within the scope of the atlas signal after the vector normalization and multiple scattering correction, K-average clustering analysis to 20 batches sulfate kind of mineral medicine is divided into seven categories, the results of the analysis method and the traditional traits identification results are basically identical. This method is simple, fast, and can be used for these minerals containing sulfate class the qualitative identification and quality control of Chinese traditional medicine.