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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.

[Objectives] By clustering analysis of tissue distribution data of brucine and strychnine in gastric ulcer model rat, the impact of Zuota on tissue distribution of basic components was studied. [Methods] Based on system clustering method of SPSS19.0 statistical analysis software, using inter-group join method and squared Euclidean distance, brucine and strychnine contents of different tissues and organs in non- Zuota group and Zuota group were taken as characteristic variables for clustering analysis, and phylogenetic tree was established. [Results] When clustering distance was 1, (i) taking brucine content as the index, there were three kinds of convergences in non-Zuota group. A1 class: skin, liver, epididymitis and jejunum; A2 class: brain and uterus; A class: testis and muscle. Brucine contents of the three classes showed a A1 < A2 < A. There were two classes of convergences in Zuota group. B1 class : jejunum, epididymis, kidney, brain, skin and uterus; B2 class: muscle and (bottom) submandibular gland. Brucine contents of the two classes showed as B1 < B2. (ii) Taing strychnine content as the index, there were three classes of convergences in non-Zuota group. C1 class: muscle, testicle and oarrum; C2 class: heart and lung; C3 class: uterus and liver. Strychnine contents of the three classes showed a C3 <C1 < C2. There were two kinds of convergences in Zuota group. D1 class : kidney and heart; D2 class : brain tissue and uterus. Strychnine contents of the two classes showed as D1 > D2. [Conclusions] When clustering distance was 1, low-content tissues and orgas firstly clustered, and its toxicological eefect(or pharmacodynamic action)was insignificant, and this kind of tissues and organs were relatively safe. A1 class and A2 class in Zuota group were merged into B1 class, in which liver was replaced by kidney. It iilustrated that Zuota could decline the toxicity of kidney, and enlarged the safe action range of brucine. Kidney and heart in C2 class were clustered into D1 class, and average strychnine content in C2 class was higher than that of D1 class. It could be deduced that Zuota had the effect of protecting heat.

To provide insights into the mechanism for the attenuate-synergistic effect of Zuota to Tibetan medicine Renqing Mangjue, a contrasted study was carried out on the pharmacokinetics of brucine and strychnine in mice plasm, which are active and toxicant ingredient in the Tibetan medicine Renqing Mangjue. LC-MS/MS was used to detect simultaneously the concentrations of brucine and strychnine in mice plasm at-different time intervals after administration parallelly and randomly, and the pharmacokinetic software Kinetica 5. 0 was selected to non-compartmental analysis (NCA) for data, and statistical analysis software SPSS 19. 0 was used for significance test on the pharmacokinetic parameters. A reliable LC-MS/MS method was established for the determination of brucine and strychnine in blood plasma, which are consistent with the requirements of the preclinical pharmacokinetic study confirmed by the methodology. The linear concentration ranges of brucine and strychnine were 0.301-104.4 µg · L(-1) (r = 0.999 5) and 0.305-106 µg · L(-1) (r = 0.999 7), respectively; The intra-day and inter-day variable coefficients were both less than 10.0% with good precision; The average extraction recoveries of brucine and strychnine were 116.23% and 112.82%, and RSD were 3.2% and 2.3% separately;The average matrix effects of brucine and strychnine were 122.48% and 116.36%, and RSD were 7.7% and 4.4%, respectively. The pharmacokinetic results showed that AUCtot of brucine and strychnine in Zuota group were both increased remarkably (P < 0.05), and the Cmax of brucine in Zuota group was about 5.25-fold higher than that of brucine in non-Zuota group (P < 0.05). The Tmax of brucine and strychnine reduced to one-eighth and one-quarter respectively compared with those in Non-Zuota group. In addition, the eliminations of brucine and strychnine in vivo were accelerated after the compatibility of Zuota. A significant difference (P < 0.05) occurred at the MRT0-t, of brucine, while the MRT0-∞ and Lz of strychnine were statistically significant upon the inspection level α = 0.1. It was found that the absorption degree of brucine and strychnine in Zuota group increased in the range of the safe dose (or concentration), while their elimination rates were accelerated, which may be one of the mechanisms for attenuate-synergistic effect of Zuota to Tibetan medicine Renqing Mangjue.

Erigeron breviscapus and erigeron multiradiatus, belonging to compositae, have been widely used as "meiduoluomi" in traditional Tibetan medicine for treatment of heat-clearing and detoxicating. However, it is difficult to distinguish them because of similarity of morphology. Therefore, the chemotaxonomy method was established. FTIR fingerprint spectra of E. breviscapus and E. multiradiatus samples from 13 geographical origins were studied. The results demonstrated that they could be divided into 2 classes with principal component analysis (PCA). The classification was well correlated to their gene, geographical origins and weather. In the same class, the chemical components are similar to each other, which can be considered as the criterion for evaluating their quality. The results showed that their infrared spectra characteristic of the same species was similar in the range of 4 000-450 cm(-1), but varied significantly for different species. The method is rapid and simple, and could be applied to evaluate the quality of this traditional medicine.

<bold>Background: </bold>Tong Luo Hua Shi (TLHS) is a new formulation of the traditional Tibetan medicine Wu-wei-gan-lu that has been used for the treatment of rheumatoid arthritis (RA) for hundreds of years in China. This study aimed to evaluate the efficacy and safety of TLHS in patients with RA.<bold>Methods: </bold>This was a randomized, double-blind, placebo-controlled, dose-finding study performed in patients with active RA from five medical centers. Patients received three doses (4.8, 3.6, or 2.4 g/day po) of TLHS or placebo (tid po) for 8 weeks. Blood sampling, physical examination, and assessment of the American College of Rheumatology (ACR) 20 % improvement (ACR20) criteria were performed before and every 2 weeks after starting treatment. The primary endpoint was the ACR20. The secondary endpoints included safety.<bold>Results: </bold>A total of 240 participants were screened and 236 patients were randomized (n = 59/group); 20 dropped out. After 8 weeks, ACR20 improvements in the TLHS 4.8 g and 3.6 g groups were significantly higher than in the placebo group (P < 0.01 and P < 0.05, respectively). ACR50 improvement in the TLHS 4.8 g group was significantly higher compared with the placebo group (P < 0.01). Symptoms of RA were significantly relieved in the TLHS groups. In the TLHS groups, insomnia (n = 1), gastroenteric reactions (n = 2), arrhythmia (n = 1), and minor hepatic lesion (n = 1) were reported; in the placebo group, hepatic dysfunction (n = 1) was reported (P = 0.878).<bold>Conclusions: </bold>TLHS improved the symptoms of patients with RA according to the ACR20. Moreover, TLHS was safe.<bold>Trial Registration: </bold>Chinese Clinical Trial Registry: ChiCTR-TRC-12003871 . Registered on 1 January 2012. [ABSTRACT FROM AUTHOR]

Flavonoids are the main components of Meconopsis integrifolia (Maxim.) Franch, which is a traditional Tibetan medicine. However, traditional chromatography separation requires a large quantity of raw M. integrifolia and is very time consuming. Herein, we applied high-speed counter-current chromatography in the separation and purification of flavonoids from the ethanol extracts of M. integrifolia flower. Ethyl acetate/n-butanol/water (2:3:5, v/v/v) was selected as the optimum solvent system to purify the four components, namely quercetin-3-O-β-d-glucopyrannosy-(1→6)-β-d-glucopyranoside (compound 1, 60 mg), quercetin 3-O-[2'''-O-acetyl-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (compound 2, 40 mg), quercetin 3-O-[3'''-O-acetyl-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (compound 3, 11 mg), and quercetin 3-O-[6'''-O-acetyl-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (compound 4, 16 mg). Among the four compounds, 3 and 4 were new acetylated flavonol diglucosides. After the high-speed counter-current chromatography separation, the purities of the four flavonol diglucosides were 98, 95, 90, and 92%, respectively. The structures of these compounds were identified by mass spectrometry and NMR spectroscopy.

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.;

Zuotais regarded as the king of Tibetan medicine. However, the major starting material ofZuotais mercury, which is one very toxic heavy metal. This has aroused serious doubts on the biosafety ofZuotacontaining drugs. In this study, we quantified the Hg contents in fourZuotasamples, 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 inZuotasamples were in the range of 566–676 mg/g. Fortunately, the release of Hg fromZuotasamples was very low in simulated gastric juice, and much lower in simulated intestinal juice. Direct contact ofZuotawith 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 toZuota. Interestingly,Zuotasamples 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 ofZuotaparticulates. Beyond the cytotoxicity, significant differences amongZuotasamples from different institutions were observed, suggesting that the preparation process ofZuotahad meaningful influence of its biosafety. The implications to the safety and clinical applications ofZuotaare discussed. [ABSTRACT FROM AUTHOR]