[Application status of metagenomics in quality research of traditional Chinese medicinal materials].

Metagenomics constructs genomic libraries by directly extracting DNA from all microorganisms from environmental samples, and studies microbial diversity and functional composition by microbiology and genomics. The metagenomic technology and genome sequencing can be used to discover the unculturable environmental microorganisms and study the natural products in the medicinal materials related to them, which overcome the limitations of microbial isolation and culture technology and is of great significance for the discovery of new genes or genomes. The quality and safety of traditional Chinese medicine materials(TCMMs) have always been a hot topic in the research on TCMMs. In addition to the intrinsic characteristics, environmental factors and human intervention also affect the quality and clinical efficacy of TCMMs. Environmental microorganisms, rhizosphere microorganisms, and endophytes play an essential role in the growth, secondary metabolism, processing, and storage of medicinal plants. As an emerging discipline, metagenomics has been used in a variety of fields. It can not only avoid the problems caused by the limited technical conditions or strict experimental requirements but also evaluate the quality of TCMMs from a new perspective. This paper introduced the application status of metagenomics in the research of TCMMs and proposed the theoretical basis and research direction for the application of metagenomics in quality research and identification of TCMMs.

Radial Pulse Wave Signals Combined with Ba-PWV for the Risk Prediction of Hypertension and the Monitoring of Its Accompanying Metabolic Risk Factors.

Our aim was to study whether radial pulse wave signals can improve the risk prediction of incident hypertension and are associated with its concomitant metabolic risk factors beyond the traditional cardiovascular risk factor Ba-PWV. By enrolling 523 Chinese subjects in this study, linear and stepwise regression analysis was performed to assess the association of radial artery pulse wave signals and Ba-PWV with blood pressure and its related metabolic risk factors such as fasting plasma glucose (FPG), total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and uric acid (UA). The area under the receiver-operating characteristic curve (AUC), net reclassification improvement (NRI), and integrated discrimination improvement (IDI) were calculated by risk assessment plot to compare the discriminative ability among models with and without radial artery pulse wave signals. After adjusting related confounding factors, radial artery pulse wave variable h 3/h 1 was selected as the sensitive influential factor for blood pressure. Moreover, a new model with h 3/h 1 had a higher AUC than the reference model without it (0.86 vs 0.84; P=0.030). And the NRI and IDI for the new model was 50.0% (P=0.017) and 3.16% (P=0.044), respectively. In addition to Ba-PWV, we found that the decrease of t 4, t 5, and h 5 might be associated with higher FPG, TC, LDL-C, and UA and lower HDL-C. This research might provide a valuable additional tool for remote wearable monitoring of radial artery pulse wave signals in hypertension risk evaluation and management.

Discovery of chemical markers for identifying species, growth mode and production area of Astragali Radix by using ultra-high-performance liquid chromatography coupled to triple quadrupole mass spectrometry.

BACKGROUND: Astragali Radix (AR) is a well-known Chinese herbal medicine. The quality of AR can be affected by many factors such as species, growth mode and production area, but there are still no chemical markers to distinguish it. PURPOSE: To explore chemical markers for improving the quality assessment of AR and discover chemical markers for identifying species, growth mode and production area of AR. METHODS: A highly sensitive, efficient and accurate method based on ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS) for simultaneous quantitative determination of 14 major chemical components (five flavonoids and nine triterpene saponins) in 94 batches of AR from China, Republic of Korea and Germany was developed for the first time. To explore chemical markers and assess changes in the contents of 14 compounds in the 94 batches of AR samples from different regions, hierarchical clustering analysis (HCA) and principal component analysis (PCA) were performed. RESULTS: Astragaloside III was not only an important chemical marker for distinguishing two species of AR, i.e.: Astragalus mongholicus and A. membranaceus, but also a potential chemical marker for the classification of cultivated and semi-wild AR. In addition, in the batches of cultivated AR, the content of isoastragaloside II and cyclocephaloside II were greater in batches from the region of Shaanxi Province than that of other Provinces in China, but the content of calycosin-7-O-ß-D-glucoside and astragaloside IV, which are the quality control markers of AR required by the Chinese Pharmacopoeia, were higher than that of other Provinces in China. In addition, the content of calycosin-7-O-ß-D-glucoside, ononin, calycosin and astragaloside I could be used to identify samples of AR collected from China, Republic of Korea and Germany. CONCLUSION: This UHPLC-QQQ-MS/MS method could be applied to the quantitative evaluation of AR and could be an important and meaningful reference to develop chemical markers for quality control of AR.

Determination of a astragaloside IV derivative LS-102 in plasma by ultra-performance liquid chromatography-tandem mass spectrometry in dog plasma and its application in a pharmacokinetic study.

BACKGROUND: Astragalosidic acid (LS-102) is a new water-soluble derivative of astragaloside IV - a major effective component isolated from the Chinese herb Astragali Radix. Our previous study showed that LS-102 exhibited potent cardiovascular activity. PURPOSE: The objective of this study was to investigate the pharmacokinetic properties of LS-102 after single-dose, oral administration in beagle dogs by developing and validating an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method. METHOD AND RESULT: The chromatographic separation was performed on a Acquity HSS C18 column (100 mm × 2.1 mm, 1.8 µm) by a gradient elution using a mobile phase consisting of water and acetonitrile at a flow rate of 0.35 ml/min. The analytes were detected with a triple quadrupole tandem mass spectrometry in multiple reaction monitoring mode. Method validation revealed a wide linearity over the range of 2.0-10,000 ng/ml together with satisfactory intra- and inter-day precision, accuracy, and recovery. Stability testing showed that LS-102 spiked into dog plasma was stable for 4 h at room temperature, for up to 2 weeks at -80 °C, and during three freeze-thaw cycles. The method was effectively and successfully applied to the pharmacokinetics of LS-102 after oral administration (5, 10 and 20 mg/kg) to beagle dogs. Peak plasma concentrations are attained within approximately 2 h after oral administration with a half-life ranging from 1.55 h to 4.49 h. The plasma concentration-time curve of LS-102 after oral administration presents the phenomenon of a double-peak absorption phase. The peak concentration and area under the concentration-time curve of LS-102 seemed to increase with the increasing doses proportionally, that suggesting linear pharmacokinetics in dogs. Meanwhile, the doxorubicin (Dox)-injured H9c2 cell model was prepared by incubating the cells in 1 µM Dox for 24 h. MTT assay and LDH release measurement showed that LS-102 protected against Dox-induced cardiomyocyte death. CONCLUSION: The obtained results may help to guide the further pre-clinical research of LS-102 as a potentially novel cardioprotective agent.

Analysis of Pulse Signals Based on Array Pulse Volume.

OBJECTIVE: To collect and analyze multi-dimensional pulse diagram features with the array sensor of a pressure profile system (PPS) and study the characteristic parameters of the new multi-dimensional pulse diagram by pulse diagram analysis technology. METHODS: The pulse signals at the Guan position of left wrist were acquired from 105 volunteers at the Shanghai University of Traditional Chinese Medicine. We obtained the pulse data using an array sensor with 3×4 channels. Three dimensional pulse diagrams were constructed for the validated pulse data, and the array pulse volume (APV) parameter was computed by a linear interpolation algorithm. The APV differences among normal pulse (NP), wiry pulse (WP) and slippery pulse (SP) were analyzed using one-way analysis of variance. The coefficients of variation (CV) were calculated for WP, SP and NP. RESULTS: The APV difference between WP and NP in the 105 volunteers was statistically significant (6.26±0.28 vs. 6.04±0.36, P=0.048), as well as the difference between WP and SP (6.26±0.28 vs. 6.07±0.46, P=0.049). However, no statistically significant difference was found between NP and SP (P=0.75). WP showed a similar CV (4.47%) to those of NP (5.96%) and SP (7.58%). CONCLUSION: The new parameter APV could differentiate between NP or SP and WP. Accordingly, APV could be considered an useful parameter for the analysis of array pulse diagrams in Chinese medicine.

A Study of Machine-Learning Classifiers for Hypertension Based on Radial Pulse Wave.

OBJECTIVE: In this study, machine learning was utilized to classify and predict pulse wave of hypertensive group and healthy group and assess the risk of hypertension by observing the dynamic change of the pulse wave and provide an objective reference for clinical application of pulse diagnosis in traditional Chinese medicine (TCM). METHOD: The basic information from 450 hypertensive cases and 479 healthy cases was collected by self-developed H20 questionnaires and pulse wave information was acquired by self-developed pulse diagnostic instrument (PDA-1). H20 questionnaires and pulse wave information were used as input variables to obtain different machine learning classification models of hypertension. This method was aimed at analyzing the influence of pulse wave on the accuracy and stability of machine learning model, as well as the feature contribution of hypertension model after removing noise by K-means. RESULT: Compared with the classification results before removing noise, the accuracy and the area under the curve (AUC) had been improved. The accuracy rates of AdaBoost, Gradient Boosting, and Random Forest (RF) were 86.41%, 86.41%, and 85.33%, respectively. AUC were 0.86, 0.86, and 0.85, respectively. The maximum accuracy of SVM increased from 79.57% to 83.15%, and the AUC stability increased from 0.79 to 0.83. In addition, the features of importance on traditional statistics and machine learning were consistent. After removing noise, the features with large changes were h1/t1, w1/t, t, w2, h2, t1, and t5 in AdaBoost and Gradient Boosting (top10). The common variables for machine learning and traditional statistics were h1/t1, h5, t, Ad, BMI, and t2. CONCLUSION: Pulse wave-based diagnostic method of hypertension has significant value in reference. In view of the feasibility of digital-pulse-wave diagnosis and dynamically evaluating hypertension, it provides the research direction and foundation for Chinese medicine in the dynamic evaluation of modern disease diagnosis and curative effect.

Tongue Image Database Construction Based on the Expert Opinions: Assessment for Individual Agreement and Methods for Expert Selection.

This study aims at introducing a method for individual agreement evaluation to identify the discordant raters from the experts' group. We exclude those experts and decide the best experts selection method, so as to improve the reliability of the constructed tongue image database based on experts' opinions. Fifty experienced experts from the TCM diagnostic field all over China were invited to give ratings for 300 randomly selected tongue images. Gwet's AC1 (first-order agreement coefficient) was used to calculate the interrater and intrarater agreement. The optimization of the interrater agreement and the disagreement score were put forward to evaluate the external consistency for individual expert. The proposed method could successfully optimize the interrater agreement. By comparing three experts' selection methods, the interrater agreement was, respectively, increased from 0.53 [0.32-0.75] for original one to 0.64 [0.39-0.80] using method A (inclusion of experts whose intrarater agreement>0.6), 0.69 [0.63-0.81] using method B (inclusion of experts whose disagreement score="0"), and 0.76 [0.67-0.83] using method C (inclusion of experts whose intrarater agreement>0.6& disagreement score="0"). In this study, we provide an estimate of external consistency for individual expert, and the comprehensive consideration of both the internal consistency and the external consistency for each expert would be superior to either one in the tongue image construction based on expert opinions.

A efficient method to identify cardioprotective components of Astragali Radix using a combination of molecularly imprinted polymers-based knockout extract and activity evaluation.

Although herb medicines have become the major source for new drug discovery, many of them are largely under-explored due to the purity-activity relationship. Efficient identification of bioactive compounds in conventional stepwise separation and isolation has not yet been elucidated. Therefore, we proposed a new separation strategy for holism understanding of herb pharmacology using molecularly imprinted polymers (MIPs). Astragali Radix (AR), important in traditional Chinese medicine, was chosen in this study for multicomponent knockout followed by bioactivity evaluation. We prepared calycosin molecularly imprinted polymers (calycosin-MIPs) which could selectively recognize flavonoid aglycons in AR. The molecular selectivity of calycosin-MIPs as a critical parameter was evaluated using the template and other high content compounds in AR. Based on it, using the calycosin-MIPs material via solid-phase extraction procedures was applied for the knockout of flavonoid aglycons in AR. Finally, hypoxia/reoxygenation model in H9c2 cells was used to evaluate the activity of the AR extract before and after knockout. The results showed that calycosin-MIPs as recognition materials for flavonoid aglycons in AR are applied in one-step separation with high selectivity and tunability. The subextract in the absence of flavonoid aglycons has been demonstrated to clarify the cardio-protective components of AR. In conclusion, this proof-of-principle study is the first one showing that molecular imprinting technology coupled with a bioassay can be used to explore the bioactive variability from the perspective of multicomponent separation of herbal medicine.

The Association of Radial Artery Pulse Wave Variables with the Pulse Wave Velocity and Echocardiographic Parameters in Hypertension.

This study aims at exploring the cardiovascular pathophysiological mechanism of TCM (traditional Chinese medicine) pulse by detecting the correlation between radial artery pulse wave variables and pulse wave velocity/echocardiographic parameters. Two hundred Chinese subjects were enrolled in this study, which were grouped into health control group, hypertension group, and hypertensive heart disease group. Physical data obtained in this study contained TCM pulse images at "Guan" position of the left hand, pulse wave velocity, and echocardiographic parameters. Linear and stepwise regression analysis was performed to assess the association of radial artery pulse wave variables with pulse wave velocity and echocardiographic parameters in the total population and in each different group. After adjusting for related confounding factors, decrease of t1, t5 and increase of h1, h3/h1 were statistically associated with arterial stiffness in the total population (P<0.05). Moreover, the correlation study in each group showed that the decrease of both t3 and h5 was also related to arterial stiffness (P<0.05). In terms of echocardiographic parameters, the height of dicrotic wave indicated by h5 was the most relevant pulse wave variable. For the health control, h5 was negatively associated with interventricular septal thickness (VST) and left ventricular posterior wall thickness (PWT) (P<0.05), while for the hypertension population and those with target-organ damage to heart, increase of h5 might be associated with decrease of ejection fraction (EF) and increase of all the remaining echocardiographic parameters especially for left ventricular end-systolic diameter (LVDs) and Left ventricular end-diastolic diameter (LVDd) (P<0.05). In conclusion, we found radial artery pulse wave variables were in association with the arterial stiffness and echocardiographic changes in hypertension, which would provide an experimental basis for cardiovascular pathophysiological mechanism of radial artery pulse wave variables.

A new approach for identification of medicinal almonds by fourier transform infrared spectroscopy and systematic clustering of characteristic peaks.

Medicinal almonds have been used for over 2 000 years and its clinical efficacy includes relieving cough and asthma. The domestic market in China is flooded with different kinds of dried almonds, such as bitter almond (Armeniacae Semen Amarum, AAS), sweet almond (Armeniacae Semen Dulce, ADS), salted almond (Armeniacae Semen Salsa, ASS), and their sulfur-fumigating products (Armeniacae Semen Sulphur Fumabat, ASFS). Wide varieties of almonds may lead to uncertain efficacy, aberrant quality, and even increased safety risk. However, the authentication method for medicinal almonds has not been reported, although imposters may lead to ineffective medical response. In the present study, Fourier transform infrared spectroscopy (FTIR) and the 2-dimensional infrared (2D-IR) spectroscopy were used to identify different almonds, which were extracted with different solvents including water, methanol, ethanol, chloroform and ethyl acetate, respectively. A new simple FTIR method was developed in the present study. According to the gradient solvent polarity, a new 2D IR method was first developed, and the commodities of almonds in China were analyzed by using the FTIR spectroscopy supported by hierarchical clustering of characteristic peaks. Moreover, 5-hydroxymethyl-2-furfural could be used as a detection index and control target in the quality control of medicinal almonds.

The Classification of Tongue Colors with Standardized Acquisition and ICC Profile Correction in Traditional Chinese Medicine.

Background and Goal. The application of digital image processing techniques and machine learning methods in tongue image classification in Traditional Chinese Medicine (TCM) has been widely studied nowadays. However, it is difficult for the outcomes to generalize because of lack of color reproducibility and image standardization. Our study aims at the exploration of tongue colors classification with a standardized tongue image acquisition process and color correction. Methods. Three traditional Chinese medical experts are chosen to identify the selected tongue pictures taken by the TDA-1 tongue imaging device in TIFF format through ICC profile correction. Then we compare the mean value of L*a*b* of different tongue colors and evaluate the effect of the tongue color classification by machine learning methods. Results. The L*a*b* values of the five tongue colors are statistically different. Random forest method has a better performance than SVM in classification. SMOTE algorithm can increase classification accuracy by solving the imbalance of the varied color samples. Conclusions. At the premise of standardized tongue acquisition and color reproduction, preliminary objectification of tongue color classification in Traditional Chinese Medicine (TCM) is feasible.

In vitro toxicity evaluations of Tibetan medicine Zuota from four institutions.

Zuota is regarded as the king of Tibetan medicine. However, the major starting material of Zuota is mercury, which is one very toxic heavy metal. This has aroused serious doubts on the biosafety of Zuota containing drugs. In this study, we quantified the Hg contents in four Zuota samples, monitored the release of Hg in simulated gastric/intestinal juice and evaluated their cytotoxicity to Caco-2 cells. Our results showed that the Hg contents in Zuota samples were in the range of 566-676 mg/g. Fortunately, the release of Hg from Zuota samples was very low in simulated gastric juice, and much lower in simulated intestinal juice. Direct contact of Zuota with Caco-2 cells led to dose-dependent cytotoxicity, including activity loss and membrane leakage. The toxicity was closely related to apoptosis, because the caspase 3/7 levels of Caco-2 cells increased after the exposure to Zuota. Interestingly, Zuota samples inhibited the oxidative stress at low concentrations, but the toxicity could be relived by antioxidants. The possible toxicity should be attributed to the cellular uptake of Zuota particulates. Beyond the cytotoxicity, significant differences among Zuota samples from different institutions were observed, suggesting that the preparation process of Zuota had meaningful influence of its biosafety. The implications to the safety and clinical applications of Zuota are discussed.

Pomegranate Seed Oil Exerts Synergistic Effects with trans-Resveratrol in a Self-nanoemulsifying Drug Delivery System.

Pomegranate seed oil (PSO) has diverse bioactivities. It was hyphothesized that if PSO were employed to construct a trans-resveratrol-loaded self-nanoemulsifying drug delivery system (RES SNEDDS-PSO), not only could PSO serve as an oil phase but also exert synergistic effects with resveratrol to yield better therapeutic outcomes. In this study, we prepared RES SNEDDS-PSO for the first time to validate that hypothesis. The anti-inflammatory and anticancer activities of RES SNEDDS-PSO were compared with another SNEDDS composed of oil phase isopropyl palmitate (RES SNEDDS-IP). The results showed that upon exposure to a 10-fold amount of water, RES SNEDDS-PSO was converted into nanoemulsions with a mean size of 44 nm. Nanoemulsions enhanced the water solubility of resveratrol by 20-fold, significantly improved resveratrol stability in intestinal fluid, and slowed the decomposition of resveratrol in water by 1-fold. An in vivo anti-infection test showed that the degree of inflammatory swelling in mice given RES SNEDDS-PSO was only 60 and 76% that of the group fed with RES SNEDDS-IP at doses of 10 and 20 mg/kg, respectively. An in vitro anticancer study showed that the inhibitory rate of RES SNEDDS-PSO against MCF-7 breast cancer cells was 2.03- and 1.24-fold that of RES SNEDDS-IP at a concentration of 12.5 and 25 µg/mL, respectively. This study demonstrated that the newly developed SNEDDS may be a prospective formulation in the functional food and clinical fields.

Syndrome Differentiation Analysis on Mars500 Data of Traditional Chinese Medicine.

Mars500 study was a psychological and physiological isolation experiment conducted by Russia, the European Space Agency, and China, in preparation for an unspecified future manned spaceflight to the planet Mars. Its intention was to yield valuable psychological and medical data on the effects of the planned long-term deep space mission. In this paper, we present data mining methods to mine medical data collected from the crew consisting of six spaceman volunteers. The synthesis of the four diagnostic methods of TCM, inspection, listening, inquiry, and palpation, is used in our syndrome differentiation. We adopt statistics method to describe the syndrome factor regular pattern of spaceman volunteers. Hybrid optimization based multilabel (HOML) is used as feature selection method and multilabel k-nearest neighbors (ML-KNN) is applied. According to the syndrome factor statistical result, we find that qi deficiency is a base syndrome pattern throughout the entire experiment process and, at the same time, there are different associated syndromes such as liver depression, spleen deficiency, dampness stagnancy, and yin deficiency, due to differences of individual situation. With feature selection, we screen out ten key factors which are essential to syndrome differentiation in TCM. The average precision of multilabel classification model reaches 80%.

[Uses of fresh herbs with antioxidant effect and prospect for population ecology reconstruction in Macau].

The average life expectancy in Macau is ranking the second in the world, the consumption of fresh medicinal plants is a profound culture in Macau. The paper focus on the distribution of the antioxidant herbs, a comprehensive investigation and analysis the amount of the plant resources was carried out. The antioxidant activity of alcohol extracts was determined by using the DPPH method, and six kinds fresh herbs with high antioxidant free radical activity were screened out. Reference to adult daily dose of vitamin C, it is calculated that the daily dose amount of fresh herbs is less than 200 g. For the expected shortage of resources and the ecological status of Macau, we give some suggestions of herbal introduction in population ecology reconstruction.

Comparative analysis of EPA and DHA in fish oil nutritional capsules by GC-MS.

BACKGROUND: Fish oil is a popular nutritional product consumed in Hong Kong. Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are the two main bioactive components responsible for the health benefits of fish oil. Market survey in Hong Kong demonstrated that various fish oil capsules with different origins and prices are sold simultaneously. However, these capsules are labelled with same ingredient levels, namely EPA 180 mg/g and DHA 120 mg/g. This situation makes the consumers very confused. To evaluate the quality of various fish oil capsules, a comparative analysis of the contents of EPA and DHA in fish oil is crucial. METHODS: A gas chromatography-mass spectrometry (GC-MS) method was developed for identification and determination of EPA and DHA in fish oil capsules. A comprehensive validation of the developed method was conducted. Ten batches of fish oil capsules samples purchased from drugstores of Hong Kong were analyzed by using the developed method. RESULTS: The present method presented good sensitivity, precision and accuracy. The limits of detection (LOD) for EPA and DHA were 0.08 ng and 0.21 ng, respectively. The relative standard deviation (RSD) values of EPA and DHA for repeatability tests were both less than 1.05%; and the recovery for accuracy test of EPA and DHA were 100.50% and 103.83%, respectively. In ten fish oil samples, the contents of EPA ranged from 39.52 mg/g to 509.16 mg/g, and the contents of DHA ranged from 35.14 mg/g to 645.70 mg/g. CONCLUSION: The present method is suitable for the quantitative analysis of EPA and DHA in fish oil capsules. There is a significant variation in the contents of the quantified components in fish oil samples, and there is not a linear relationship between price and contents of EPA and DHA. Strict supervision of the labelling of the fish oil capsules is urgently needed.

[UPLC fingerprint spectra for discrimination of Aucklandiae radix and Vladimiriae radix].

It's difficult to identify Aucklandiae Radix and Vladimiriae Radix because of their similar composition. In this paper, UPLC method was used to establish their UPLC fingerprint to identify them with the mobile of acetonitrile -0. 05% phosphoric acid water solution by gradient elution at the detection wavelength of 238 nm. Clustering analysis and principal components analysis showed that Vladimiriae Radix was significantly different from Aucklandiae Radix. Eight common peaks and twelve common peaks were defined respectively in Aucklandiae Radix and Vladimiriae Radix herbs by fingerprint analysis. Six of them were identified as syringoside, chlorogenic acid, isochlorogenic acid A, isochlorogenic acid B, costunolide and dehydrocostuslactone by comparing with standard references. There are four peaks in all of Vladimiriae Radix samples and in none of Aucklandiae Radix samples. So UPLC fingerprint can be used to identify these two herbs.

Hepatoprotective activity of Gentiana veitchiorum Hemsl. against carbon tetrachloride-induced hepatotoxicity in mice.

AIM: To study the hepatoprotective effect of methanol extract of Gentiana veitchiorum (MGV) against CCl4-induced oxidative stress and liver injury in mice. METHOD: The acute hepatic model was developed by injection of 20% CCl4 in mice. ICR mice were divided into six groups, including control, CCl4, CCl4(+) silymarin, and CCl4(+) MGV (100, 200, and 400 mg·kg(-1)) groups. Hepatic enzymes including AST, ALT and ALP levels in serum, and antioxidant enzymes, including SOD, CAT and GPX activity in liver tissue, were determined. Histopathological examination and Western blot analysis were performed. RESULTS: Oral administration of MGV at 200 and 400 mg·kg(-1) for 15 days dose-dependently inhibited the serum elevations of AST, ALT, and ALP, and recovered the reduction of SOD, CAT, and GPX in liver tissue. Hematoxylin and eosin staining examination performed in liver tissues suggested that MGV treatment ameliorated histopathological changes in CCl4-induced mice. Western blotting analysis implied that MGV increased HO-1 expression and recovered TNF-α alternation. CONCLUSION: G. veitchiorum can protect the liver against CCl4-induced damage in mice, and this hepatoprotective effect was due at least in part to its ability through scavenging CCl4-associated free radical activities. The study provided in vivo evidence that G. veitchiorum can be used as a safe, cheap, and effective agent to reduce acute liver damage, supporting its folk medicine use.

Characterization of tibetan medicine zuota by multiple techniques.

Zuota is regarded as the king of Tibetan medicine. However, due to the confidentiality of this precious medicine, the scientific characterization of Zuota is very scarce, which limits the pharmacology and biosafety studies of Zuota. Herein, we collected four different Zuota samples from Tibet, Qinghai, Gansu, and Sichuan and characterized them by multiple techniques. Our results showed that Zuota was mainly an inorganic mixture of HgS, sulfur, and graphite. Morphologically, Zuota samples were composed of nanoparticles, which further aggregated into microsized particles. Chemically, the majorities of Zuota were S and Hg (in the forms of HgS and pure sulfur). All samples contained pure sulfur with orthorhombic crystalline. Zuota from Qinghai province had different HgS crystalline, namely, hexagonal crystalline. The others were all face-centered cubic crystalline. Carbon in Zuota NPs was in the form of graphite. The implication to future studies of Zuota was discussed.

[Spectrum-effect correlation analysis of traditional Tibetan medicine "Morina nepalensis" on nitric oxide production inhibition].

OBJECTIVE: To establish an HPLC-ELSD fingerprint of the whole herbs of Morina nepalensis and perform the correlation analysis of chemical components of the herb and nitric oxide (NO) production inhibition. METHOD: HPLC-ELSD assay was performed to evaluate 10 batches of M. nepalensis herbs. The chromatographic conditions were as following: Eclipse XDB C18 column (4.6 mm x 150 mm, 5 microm), water (A) and acetonitrile (B) as a gradient mobile phases, flow rate 1.0 mL x min(-1), and column temperature at 35 degress C. Evaporative light-detection conditions: atomization temperature at 104 degrees C, the flow rate of N2 2.8 L x min(-1) and 10 microL sample injection. Chromatographic fingerprint was developed, and the inhibition activity of production of NO in lipopolysaccharide-induced macrophages was also analyzed. The similarity and correlation analysis between the HPLC-ELSD fingerprints and NO production inhibition were carried out by PLS method. RESULT: The common mode for M. nepalensis herb fingerprint was established, including 15 common characteristic peaks. Among them, 7 peaks were positively correlated with the NO production inhibition. According to the assessment on the similarity of 10 batches of samples, a similarity of over 0.90 were shown in HPLC-ELSD fingerprint and all samples were separated into two groups. CONCLUSION: This method can be used to assess the quality of M. nepalensis, which provides a reliable method for scientific assessment and quality control.