Hepatotoxicity and mechanism of Rhododendri Mollis Flos based on zebrafish model / 中国中药杂志

This study used the zebrafish model to explore the hepatotoxicity of Rhododendri Mollis Flos(RMF). The mortality was calculated according to the number of the survival of zebrafish larvae 4 days after fertilization under different concentration of RMF, and the dose-toxicity curve was fitted to preliminarily evaluate the toxicity of RMF. The liver phenotypes under the sublethal concentration of RMF in the treatment group and the blank control group were observed by hematoxylin-eosin(HE) staining and acridine orange(AO) staining. Meanwhile, the activities of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were determined to confirm the hepatotoxicity of RMF. Real-time quantitative polymerase chain reaction(real-time PCR) and Western blot were used to determine the expressions of genes and proteins in zebrafish larvae. Gas chromatography time-of-flight mass spectrometry(GC-TOF-MS) was used to conduct untargeted metabolomics testing to explore the mechanism. The results showed that the toxicity of RMF to zebrafish larvae was dose-dependent, with 1 100 μg·mL~(-1) of the absolute lethal concentration and 448 μg·mL~(-1) of sublethal concentration. The hepatocyte apoptosis and degeneration appeared in the zebrafish larvae under the sublethal concentration of RMF. The content of ALT and AST in zebrafish larvae at the end of the experiment was significantly increased in a dose-dependent manner. Under the sublethal concentration, the expressions of genes and proteins related to apoptosis in zebrafish larvae were significantly increased as compared with the blank control group. The results of untargeted metabolomics showed that the important metabolites related to the he-patotoxicity of RMF were mainly enriched in alanine, aspartic acid, glutamic acid, and other pathways. In conclusion, it is inferred that RMF has certain hepatotoxicity to zebrafish larvae, and its mechanism may be related to apoptosis.

Identification of geographical origins of Cordyceps based on data of amino acids with self-organizing map neural network / 中国中药杂志

In this study, data of amino acids of Cordyceps samples from Qinghai and Tibet was analyzed with self-organizing map neural network. A model of XY-Fused network was established with the content of 8 major amino acids and total amino acids for the identification of geographical origins of Cordyceps from Qinghai and Tibet. It had the prediction accuracy of 83.3% for the test set. In addition, data mining indicated that methionine was a special kind of amino acid in Cordyceps which could serve as a marker to identify its geographical origins. On this basis, the content ratio of methionine to total amino acids was proposed to be a quantifiable indicator to distinguish Cordyceps from Qinghai and Tibet.

Overview of metalomics research and its application prospect in mineral Chinese medicine research / 中国中药杂志

Metallomics is a frontier interdisciplinary subject at its vigorous development stage. Its goal is to systematically study the content, distribution, chemical species, structural characteristics and functions of metal elements in biological system. It is also a comprehensive discipline to study the existing state and function of free or complex metal elements in life. Metallomics is an ideal tool to study the biological behavior of inorganic elements, which can be used to solve many problems in the research of mineral Chinese medicine(MCM). It provides a strong theoretical basis and technical support for the research of MCM. Its theory and methods provide re-ference and enlightenment for the in-depth study of MCM, and also provide new ideas and open up new ways for the research of MCM. The application of metallomics theory and methods in the research of MCM is of great significance to reveal the material basis and mec-hanism of MCM, promote the process of basic research on MCM, fully exploit and utilize medicinal mineral resources and carry forward the traditional MCM treasure in China. In this paper, we introduced the concept, academic development, research content and research methods of metallomics, and discussed the application prospects of metallomics in the analysis of inorganic element composition characteristics and quality control, material basis and mechanism of MCM, so as to provide reference for further researches on MCM.

Study on liver diseases with zebrafish as an important tool / 中国中药杂志

With the increasing incidence of hepatobiliary diseases, it is particularly important to understand the role of molecular, cellular and physiological factors in the clinical diagnosis and treatment with traditional Chinese medicine(TCM) in the development of liver disease. Appropriate animal models can help us identify the possible mechanisms of relevant diseases. Danio rerio(zebrafish) model was traditionally used to study embryonic development, and has been gradually used in screening and evaluation of liver diseases and relevant drug in recent years. Zebrafish embryos develop rapidly and the digestive organs of 5-day-old juvenile fish are all mature. At this stage, they may develop hepatobiliary diseases induced by developmental defects or compounds. Zebrafish liver is similar to human liver in cell composition, function, signal transduction, response to injury and cell process mediating liver disease. Furthermore, due to the high conservation of genes and proteins between humans and zebrafish, zebrafish becomes an alternative system for studying basic mechanisms of liver disease. Therefore, genetic screening could be performed to identify new genes involving specific disease processes, and chemical screening could be made for drugs in specific processes. This paper briefly introduced the experimental properties of zebrafish as model system, emphasized the study progress of zebrafish models for pathological mechanism of liver diseases, especially fatty liver, and drug screening and evaluation, so as to provide ideas and techniques for the future liver toxicity assessment of TCM.

Antidiarrheal Effect of Limonitum with Different Mineral Genesis / 中国实验方剂学杂志

Objective: To investigate the antidiarrheal effect and the gastrointestinal motility of two kinds of Limonitum with different mineral genesis, so as to select the optimal mineral genesis, and provide a theoretical basis for clinical use. Method: The selected research samples were leaching type and sedimentary type Limonitum with different mineral genesis, including powder, water decoction and dregs. In the experiment, castor oil (0.2 mL/10 g) was used to induce diarrhea, loperamide (4 mg·kg-1) was used for positive control, and then the antidiarrheal activity and the gastrointestinal motility were observed. Result: In the castor oil-induced diarrhea experiment, the diarrhea index of the water decoction (2.5,5,10 g·kg-1) and the dregs (2.5 g·kg-1) of leaching type Limonitum significantly reduced (P-1) of leaching type limonitum and the dregs (2.5,5,10 g·kg-1) of the leaching type Limonitum were significant different at the first defecation (P-1) and the water decoction (2.5 g·kg-1) of leaching type Limonitum, and the total feces weight of the mice were significantly decreased (P-1), the dregs (2.5,5,10 g·kg-1) of the sedimentary, the powder (2.5,5 g·kg-1), the water decoction (2.5,5,10 g·kg-1) and the dregs (10 g·kg-1) of leaching type Limonitum were significantly lower than those of the model group on the intestinal motility, with charcoal as a marker (P-1) of leaching type Limonitum was significantly higher than that of the model group on the content of small intestine contents (P-1) of sedimentary type limonitum, and the water decoction (2.5,5 g·kg-1) of leaching type Limonitum were significantly lower than those of the model group on gastric residual rate (P-1), the dregs (5 g·kg-1) of sedimentary type Limonitum were significantly reduced on the water content of large intestine compared with the model group (P-1 of Limonitum can inhibit the contraction of isolated ileum induced by acetylcholine in rabbits except 0.003 3 g·mL-1 water decoction of leaching type. Conclusion: Mineral medicine Limonitum has a certain therapeutic effect in castor oil-induced diarrhea. Leaching type limonitum has the best antidiarrheal effect.

SEM and XRD Identification of Mineral Chinese Medicine Alumen, Calcined Alumen, and Their Processed Products / 中国实验方剂学杂志

Objective: Ammonium alum is a common counterfeit of Alumen,and the processed product of ammonium alum is a common counterfeits of calcined Alumen. This paper aims to establish a method for identifying Alumen,calcined Alumen,ammonium alum and their processed products. Method: The samples were analyzed by scanning electron microscope (SEM) and X ray diffraction (XRD) in this paper. Result: Ammonium alum and Alumen showed obvious changes in morphology after processing. Both Alumen and ammonium alum showed obvious differences in morphology at×250 and×1 000 times microscope. Alumen presented irregular fragments,clear edge corners,smooth surface,scattered irregular small particles,occasional holes and longitudinal edges. Ammonium alum presented irregular clumps,blunt edges,not obvious edges and corners,uneven surface,scattered smaller and round-like particles. The difference in morphology was not obvious at×250 times microscope between Alumen and ammonium alum processed products. While at×1 000 times,the surface of calcined Alumen was uneven with coarse particles; the surface of counterfeit calcined Alumen was flat,and the coarse particle characteristics were not obvious. XRD can be used to rapidly and accurately identify the primary phase of Alumen,calcined Alumen,ammonium alum and ammonium alum processed products:KAl(SO4)2·12H2O,NH4Al(SO4)2·12H2O,KAl(SO4)2,and NH4Al(SO4)2 respectively, with 2θ angle characteristic value of 23,12,22 and 5 respectively for XRD peak. Conclusion: SEM and XRD techniques can be used for the identification of Alumen,calcined Alumen,ammonium alum and their counterfeit products.

Research on Differential Feature Elements and Characteristic Spectrum of Inorganic Elements Between Calcined Alumen and Its Counterfeit Processed Products Based on ICP-OES/ICP-MS / 中国实验方剂学杂志

Objective: To provide the new quality control means for Alumen by investigating the elemental differences between calcined Alumen and its counterfeit processed products of ammonium alum, and establishing their characteristic chromatogram. Method: The contents of 22 inorganic elements both in calcined Alumen and processed products of ammonium alum were determined by means of inductively coupled plasma (ICP)-optical emission spectrometer-mass spectrometry (ICP-OES/ICP-MS),SPSS 16.0 was used for cluster analysis (CA) while SIMCA-P 13.0 with t-test and Rank-Sum test was used to identify the differential inorganic elements. In addition, the characteristic spectrum of the inorganic elements for calcined Alumen and counterfeit calcined alumen were established. Result: Calcined Alumen had highest contents of K and Al while counterfeit calcined Alumen has highest contents of Al and Fe;Cr,Sr,and Mn contents in calcined Alumen were relatively higher,while Mn,Ti,and Ga contents in processed products of ammonium alum were relatively higher. The content of K in calcined Alumen was about 205 times of that of counterfeit products. On the contrary,the average contents of Fe,Ti,Mn and Ga in counterfeit products of ammonium alum were much higher than those in calcined Alumen,33,46,38, 27 times, respectively. A total of 18 samples were clustered into two categories in CA:calcined Alumen and processed products of ammonium alum. 18 inorganic elements showed significant difference in contents(PConclusion: This method can be used for quality control of calcined Alumen.

Accumulation and Toxicity of Mercury in Zhuhong Ointment in Rats with Damaged Skin Model / 中国实验方剂学杂志

Objective: To study the accumulation of mercury in liver, kidney and brain of rats and its toxicity on liver and kidney after 4 weeks of administration of different doses of Zhuhong ointment, in order to provide data reference for the safe clinical use of Zhuhong ointment. Method: Forty-four Sprague-Dawley (SD) rats were randomly divided into 4 groups:control group, normal-dose group (1.875 mg·kg-1), medium-dose group (37.5 mg·kg-1), and high-dose group (75 mg·kg-1). After transdermal administration for consecutive 4 weeks, the mercury content in the urine, blood, liver, kidney and brain of the rats was measured. In addition, serum alanine aminotransferase (ALT), serum aspartate aminotransferase (AST), serum urea nitrogen (BUN), and serum creatinine (SCr), urine β2-miSCroglobulin (β2-BMG) and urine N-acetyl-beta-D glucosidase (NAG) contents were measured, and pathological morphology changes of liver and kidney were observed. Result:Compared with the control group, the levels of blood mercury and urine mercury in Zhuhong ointment groups showed significant increases after administration for 4 weeks(PPPPPPConclusion:Zhuhong ointment is not toxic at the normal dose, but long-term use can lead to the accumulation of mercury in liver, kidney and bra, which causes liver and kidney toxicity. This study did not find a more sensitive indicator of liver and kidney toxicity than liver and kidney pathology. However, because the rising levels of urinary mercury and blood mercury may predict toxicity, the relationship between mercury exposure and toxicity could be further studied. This study provides a reference for the rational use and toxicity monitoring of Zhuhong ointment.

Research progress in mineral Chinese medicine realgar / 中国中药杂志

Realgar is a mineral traditional medicine with definite efficacy. The function of realgar is detoxicating, insecticiding, eliminating dampness and phlegm, etc. It is widely applied in clinical practice by compatibility medicines. However, the safety and scientificalness of clinical application are questioned because of the toxic effect caused by arsenic compounds. At present, there are still many problems in the research of realgar, which are mainly manifested in three areas: the expression of main components and effective substances are inconsistent; the anti-tumor mechanism is difficult to explain at the molecular level; the mechanism of compatibility is not clear. As a result, realgar and realgar-containing Chinese patent medicines are frequently prohibited from entering the international market, and the reputation of traditional Chinese medicine is also damaged. This paper would analyze the research status of realgar at home and abroad as well as its problems from its main components, effective substances, anti-tumor mechanism and compatibility mechanism. In view of these difficulties, quantum chemical calculation method is proposed to solve them, so as to make up for the shortcomings and limitations of experimental technology and experimental conditions, reduce the cost of realgar research and improve research efficiency. Moreover, it provides inspiration for research of other mineral medicine.

The molecular identification of Bupleurum medicinal species and the quality investigation of Bupleuri Radix / 药学学报

Bupleuri Radix is one of the most frequently used herbal medicines in China with a 2000-year medicinal history. However, the use of Bupleuri Radix is very confused. Twenty-five species and eight varieties of Bupleurum have been used as Bupleuri Radix in different regions of China. It is very difficult to identify these Bupleurum species using traditional morphological method. In order to establish a fast and effective method to identify these Bupleurum species, we collected 168 Bupleurum medicinal plants from 14 populations of 9 provinces, and amplified their ITS sequences. 168 ITS sequences with a full length of 600-606 bp were obtained. DNAMAN analyzing results showed that 86 variable sites were present in these sequences and 19 haplotypes (TH1-TH19) were determined. After calculating K2P distance and analyzing an NJ tree, we established a molecular identification method based on ITS sequence. Using this method,52 samples of Bupleuri Radix were identified successfully. Furthermore, we tested saikosaponin a, c, d contents in these Bupleuri Radix by HPLC and analyzed the results by ANOVA and LSD T test to evaluate the quality of Bupleuri Radix. This method is significant for effective identification of Bupleurum medicinal plants, and quality control of Bupleuri Radix in the market.

The molecular identification of licorice species and the quality evaluation of licorice slices / 药学学报

Licorice is one of the most common herbs in traditional Chinese medicine, and classified as top grade in Shen Nong Ben Cao Jing. There are three different original plants of licorice stipulated in Chinese Pharmacopeia, Glycyrrhiza uralensis Fisch., Glycyrrhiza glabra L., and Glycyrrhiza inflata Bat. However, previous investigation showed that the pharmacodynamic effects of the three licorices were quite different. It is very difficult to identify them by the classical identification methods. In order to establish a fast and effective identification method, we collected 240 licorice plants from 21 populations of 7 provinces, and amplified their ITS and psbA-trnH sequences. ITS sequences with a full length of 616 bp and psbA-trnH sequences with a full length of 389 bp were obtained separately. Using DNAMAN to analyze these sequences, 4 variable sites were found in ITS sequences and 2 ITS haplotypes were determined, and 3 variable sites were found in psbA-trnH sequences and 4 psbA-trnH haplotypes were determined. With the combination analysis of ITS and psbA-trnH sequences, the molecular identification method of original licorice was established. Using this method, 40 samples of licorice slices collected from 4 main herbal material markets in China were identified successfully. Furthermore, the contents of 2 triterpenes, 18α-glycyrrhizic acid and 18β-glycyrrhizic acid, and 4 flavonoids, liquiritin, isoliquiritin, liquiritigenin, and isoliquiritigenin in these licorice pieces were examined by HPLC and the results were analyzed using SPSS 21.0. This study provides a new method in identification of licorice, which may serve as a guideline for quality control of licorice slices.

Toxicity of mineral Chinese medicines containing mercury element / 中国中药杂志

Mineral Chinese medicine is the distinctive part of the Chinese traditional medicine. The mineral Chinese medicines containing mercury elements such as cinnabaris, calomelas and hydrargyri oxydum rubrum are widely applied in the clinical conditions because of their efficacy of sedative, sterilization, removing necrotic tissue and promoting granulation. However, the rationality and security of clinical application are questioned because of the toxic effect caused by mercury compounds. This paper would summarize the efficacy of the mineral Chinese medicines containing mercury element, as well as their hepatotoxicity, nephrotoxicity, embryotoxicity, and neurotoxicity effect and mechanisms. Improper usage or high dose of the mineral Chinese medicines containing mercury element would cause acute hepatotoxicity. Cinnabaris, calomelas and hydrargyri oxydum rubrum may lead to chronic hepatotoxicity, nephrotoxicity, embryotoxicity and neurotoxicity when they were applied externally to the skin for long-term use. In addition to the accumulation of mercury elements in the tissues and organs, the species and forms of mercury compounds absorbed into the body in different ways, should be also studied in order to understand the toxicity of the mineral Chinese medicines containing mercury element. Meanwhile the dose and period of treatment shall be also considered in order to provide the references for rational and safe clinical application of the mineral Chinese medicines containing mercury element.

Advances of toxicity evaluation of cinnabar and compatibility necessity analysis / 中国中药杂志

Cinnabars are widely used in the clinic and shows unique efficacy, and it has been used in treating diseases for thousands of years either single-use or combination with other traditional Chinese medicines. Cinnabar mainly contains mercury, which displays obvious hepatotoxicity and nephrotoxicity. The safety of cinnabar and its rational use caused extensive attention. However, there are some misunderstanding that cinnabar shows equivalent toxicity to the toxicity of mercury compounds such as mercuric chloride and methylmercury in safety evaluation of cinnabar, which in our view exaggerates the toxicity. For traditional Chinese medicine containing cinnabar, some researchers questioned the medicinal value of cinnabar, and even proposed to remove cinnabar in order to avoid possible poisoning. This review reported adverse events, summarized its toxicity characteristics, mechanism and the medicinal value in preparations of traditional Chinese medicine. The significance of this paper is to further understand the toxicity of cinnabar and reduce its side effect in clinical application, and to provide a scientific basis for reasonable clinical application and a scientific understanding of cinnabar.

Studies of pattern recognition of fingerprint profile of cattle bile powder and determination of multi-component in it / 中国中药杂志

An HPLC-ELSD method with good specificity and good accuracy was used for the studies of fingerprint and quantification of multi-components for cattle bile powder. The chromatographic analysis was carried out on a Phenomenex Gemini C₁₈ column (4.6 mm×250 mm, 5 μm) with a column temperature of 40 ℃ and a liquid flow-rate of 1.0 mL•min⁻¹ using 10 mmol ammonium acetate solution and acetonitrile as the mobile phase with a linear gradient. An ELSD was used with a nitrogen flow-rate of 2.8 L•h⁻¹, at a drift tube temperature of 110 ℃. The average contents of glycocholic acid, glycodeoxycholic acid, taurocholic acid, taurodeoxycholic acid were (25.2±17.0)%, (4.1±3.4)%, (24.5±20.0)% and (5.2±3.8)% respectively, and the total content of the four bile acids was (59.0±26.0)%. Beyond that, the preprocessing and pattern recognition analysis of the chromatographic fingerprints of samples were applied with chemometric method. The results of this chemometric analysis indicated that the samples from market and self-made samples were different signally, and four regions were noteworthy due to their great impact with poor chromatographic signal. All in one, because this HPLC-ELSD method was simple and accurate, it was suitable for the quality assessment and quality control of cattle bile powder and could be the technological base for its standard perfection.

Chemical constituents from safflower injection and their bioactivity / 中国中药杂志

The chemical constituents of Safflower injection were isolated and purified by polyamide, silica gel, Sephadex LH-20, ODS column chromatographies and preparative HPLC. As a result, sixteen compounds have been isolated. Based on the spectral data analysis, their structures were elucidated as scutellarin (1), kaempferol-3-O-β-rutinoside(2), hydroxysafflor yellow A(3), rutin (4), coumalic acid(5), adenosine(6), syringoside(7), (3E)-4-(4'-hydroxyphenyl)-3-buten-2-one(8), (8Z)-decaene-4, 6-diyne-1-Oβ-D-glucopyranoside(9), 4-hydroxybenzaldehyde (10), (2E, 8E) -tetradecadiene-4, 6-diyne-1, 12, 14-triol-1-O-β-D-glucopyranoside (11), kaem-pferol-3-O-β-sophorose (12), uridine (13), roseoside (14), cinnamic acid (15), and kaempferol (16). Compounds 1,2,7,9,11 and 12 were isolated from the Safflower injection for the first time. The anti-platelet aggregation activities of the isolated compounds were assayed. The results indicated all tested compounds exhibited potent activity except for 5, while 2, 3, 9 and 12 showed strong activity against platelet aggregation.

Macroscopic and microscopic characteristics of Chinese caterpillar fungus / 药学学报

Chinese Caterpillar Fungus (CCF) is one of the rare Chinese traditional drugs. As the resource is reducing sharply, the price is rising higher and higher, and there have been much more adulterants in the markets, but until now we don't have a scientific and accurate research on the identification study for this drug. On the basis of resource investigation, during the study of the samples collected by ourselves and the specimens stored in the museum, using the macroscopic and microscopic methods, referring to the literatures of entomology, emphasizing on the characteristics of polypide part, we have studied this species in detail of the macroscopic characters such as the insertion position of the stroma part, the annulations and segments of the caterpillar, the abdominal leg, the pinaculum, and the microscopic characters of the body wall; firstly added the microscopic character of the crotchets on the planta of abdominal leg. The result turned out that the characters which we have studied are regular and stable, and it have laid the foundation for the powder products and patent medicines which have used the crude drug of CCF.

Study on saponins in shengmai injection / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the saponin in Shengmai injection.</p><p><b>METHOD</b>On the basic of studing the chemical constituents of red ginseng and Shengmai injection, 20 compositions had been identifided by LC-MS/MS.</p><p><b>RESULT</b>Twenty identifided compositions were the common components of Shengmai injection and red ginseng extracts.</p><p><b>CONCLUSION</b>The analytical method for saponins in Shengmai injection was established which could be used as the basis for further study and quality control.</p>

Chemical constituents of surface layer of Poria cocos and their pharmacological properties (I) / 中国中药杂志

The surface layer of the sclerotia of Poria cocos, named Fu-Ling-Pi, is used as a diuretic in traditional Chinese medicine to treat edema and urinary dysfunction. Recent studies have showed that the triterpenes (lanostane and 3,4-secolanostane skeletons) and polysaccharides (beta-pachyman) are the main components of Fu-Ling-Pi and they exhibited various biological activities, such as anti-tumor, antibacterial and antioxidant, etc. This review was focused on the chemistry, pharmacology, and clinical uses of this drug and it may provide scientific foundation for further development and utilization of Fulingpi.

Study of determination method for heavy metals and harmful elements residues in four traditional Chinese medicine injections / 中国中药杂志

Methods for determination of heavy metals and harmful residues in traditional Chinese medicine injection were established. Graphite furnace atomic absorption spectrometry was used for determination of lead, cadmium and copper, atomic fluorescence spectrometry for arsenic and mercury. The preprocessing method was optimized. The average recoveries of 5 elements were between 91% and 112% while the precisions were less than 2%. The determination limit of lead, cadmium, copper, arsenic and mercury were 0.28, 0.014, 0.49, 0.19, 0.061 microg x L(-1), respectively. The proposed method was simple, sensitive, accurate and reliable, and could be used widely.

Studies on chemical constituent and quantitative determination of Mallotus apelta / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents and establish a quantitative method of Mallotus apelta.</p><p><b>METHOD</b>Compound was isolated by silica gel, Sephadex LH-20 column chromatography and Pre-HPLC chromatography. Its structure was identified by physicochemical properties and spectral evidences. The content of M. apelta was determined by HPLC. Chromatographic conditions included Inertsil ODS-3 C18 column (4.6 mm x 150 mm, 5 microm) and the mobile phase consisting of a mixture of methanol-water (24:76). The detection wavelength was set at 335 nm.</p><p><b>RESULT</b>One compound was isolated from n-butanol extract of the M. apelta and its structure was identified as vicenin II. The calibration cure was linear in the range of 0.053-10.60 microg (r = 0.9999), the average recovery was 99. 32%, RSD 1.82% (n = 6).</p><p><b>CONCLUSION</b>The compound was isolated from this plant for the first time. The method to determine the content of vicenin II by HPLC was established for the first time. This method is simple, accurate and reliable.</p>