Research status of Bovis Calculus and relevant Chinese patent medicines based on bibliometric analysis / 中国中药杂志

With scarce resources, natural Bovis Calculus is expensive and hard to meet clinical demand. At the moment, four kinds of Bovis Calculus are available on the market: the natural product, in vitro cultured product, synthesized product, and the product formed in cow after manual intervention. In this study, papers on the four kinds of Bovis Calculus products and relevant Chinese patent medicines were searched from Web of Science, PubMed, and China National Knowledge Infrastructure(CNKI). CiteSpace, citexs AI, and CNKI were employed for bibliometric analysis and knowledge map analysis. On this basis, the status, trend, and focuses of research on Bovis Calculus and relevant Chinese patent medicines were summarized. The results suggested overall slow development in the research on Bovis Calculus and relevant Chinese patent medicines with three typical growth stages. It is consistent with the development of Bovis Calculus substitutes and the national policy for the development of traditional Chinese medicine. At the moment, the research on Bovis Calculus and relevant Chinese patent medicines has been on the rise. In recent years, there has been an explosion of research on them, particularly the quality control of Bovis Calculus and the Chinese patent medicines, the pharmacological efficacy of Chinese patent medicines, such as Angong Niuhuang Pills, and the comparison of the quality of various Bovis Calculus products. However, there is a paucity of research on the pharmacological efficacy and the mechanism of Bovis Calculus. This medicinal and the relevant Chinese patent medicines have been studied from diverse perspectives and China becomes outstanding in this research field. However, it is still necessary to reveal the chemical composition, pharmacological efficacy, and mechanism through multi-dimensional deep research.

Effect and mechanism of Bovis Calculus on ulcerative colitis by inhibiting IL-17/IL-17RA/Act1 signaling pathway / 中国中药杂志

This study aimed to elucidate the effect and underlying mechanism of Bovis Calculus in the treatment of ulcerative colitis(UC) through network pharmacological prediction and animal experimental verification. Databases such as BATMAN-TCM were used to mine the potential targets of Bovis Calculus against UC, and the pathway enrichment analysis was conducted. Seventy healthy C57BL/6J mice were randomly divided into a blank group, a model group, a solvent model(2% polysorbate 80) group, a salazosulfapyridine(SASP, 0.40 g·kg~(-1)) group, and high-, medium-, and low-dose Bovis Calculus Sativus(BCS, 0.20, 0.10, and 0.05 g·kg~(-1)) groups according to the body weight. The UC model was established in mice by drinking 3% dextran sulfate sodium(DSS) solution for 7 days. The mice in the groups with drug intervention received corresponding drugs for 3 days before modeling by gavage, and continued to take drugs for 7 days while modeling(continuous administration for 10 days). During the experiment, the body weight of mice was observed, and the disease activity index(DAI) score was recorded. After 7 days of modeling, the colon length was mea-sured, and the pathological changes in colon tissues were observed by hematoxylin-eosin(HE) staining. The levels of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6(IL-6), and interleukin-17(IL-17) in colon tissues of mice were detected by enzyme-linked immunosorbent assay(ELISA). The mRNA expression of IL-17, IL-17RA, Act1, TRAF2, TRAF5, TNF-α, IL-6, IL-1β, CXCL1, CXCL2, and CXCL10 was evaluated by real-time polymerase chain reaction(RT-PCR). The protein expression of IL-17, IL-17RA, Act1, p-p38 MAPK, and p-ERK1/2 was investigated by Western blot. The results of network pharmacological prediction showed that Bovis Calculus might play a therapeutic role through the IL-17 signaling pathway and the TNF signaling pathway. As revealed by the results of animal experiments, on the 10th day of drug administration, compared with the solvent model group, all the BCS groups showed significantly increased body weight, decreased DAI score, increased colon length, improved pathological damage of colon mucosa, and significantly inhibited expression of TNF-α,IL-6,IL-1β, and IL-17 in colon tissues. The high-dose BCS(0.20 g·kg~(-1)) could significantly reduce the mRNA expression levels of IL-17, Act1, TRAF2, TRAF5, TNF-α, IL-6, IL-1β, CXCL1, and CXCL2 in colon tissues of UC model mice, tend to down-regulate mRNA expression levels of IL-17RA and CXCL10, significantly inhibit the protein expression of IL-17RA,Act1,and p-ERK1/2, and tend to decrease the protein expression of IL-17 and p-p38 MAPK. This study, for the first time from the whole-organ-tissue-molecular level, reveals that BCS may reduce the expression of pro-inflammatory cytokines and chemokines by inhibiting the IL-17/IL-17RA/Act1 signaling pathway, thereby improving the inflammatory injury of colon tissues in DSS-induced UC mice and exerting the effect of clearing heat and removing toxins.

Investigation of chemical profile of bile acids and determination of them in cultured Bovis Calculus with UPLC-QDA / 中国中药杂志

In this study, 10 batches of samples of cultured Bovis Calculus(cow-bezoar) were determined for the investigation of chemical profile of bile acids with the UPLC-QDA method established. The results showed that nine common bile acids, cholic acid(CA), deoxycholic acid(DCA), chenodeoxycholic acid(CDCA), taurocholic acid(TCA), taurodeoxycholic acid(TDCA), taurochenodeoxycholic acid(TCDCA), glycocholic acid(GCA), glycodeoxycholic acid(GDCA) and glycochenodeoxycholic acid(GCDCA), were founded in samples and 7 bile acids were quantified except GDCA and GCDCA. In these samples, unconjugated bile acids, the major type of bile acids, accounted for more than 97% of all types of bile acids. As for unconjugated bile acids, CA was the most major bile acid in cultured cow-bezoar and it was about twice as much as DCA. The relative low-cost method established in the current study is accurate, rapid and sensitive, which is suitable for the studies of other drugs from animal bile.

Overall quality control of Jinzhen Oral Liquid based on HPLC-UVD-ELSD fingerprint and simultaneous determination of 13 main representative components / 中草药

Objective: To establish the fingerprint of Jinzhen Oral Liquid (JOL) by HPLC-UVD-ELSD and determine the main 13 representative components (gallic acid, liquiritin, aloe-emodin-8-O-β-D-glucopyranoside, liquiritigenin, baicalin, chrysin-7-O-β-D- glucoronide, oroxyloside, wogonoside, chrysophal-1-O-β-D-glucopyranoside, chrysophal-8-O-β-D-glucopyranoside, rhein, glycyrrhizic acid, hyodeoxycholic acid and cholic acid) simultaneously, in order to provide reference for the overall quality control of JOL. Methods: The separation was developed on Cosmosil-C18 column (250 mm × 4.6 mm, 5 μm) by gradient elution with methanol-water [containing 0.1% formic acid] at 254 nm, the temperature of drift tube was maintained at 115 ℃ and the carrier gas flow rate was 2.0 L/min. An HPLC-UVD-ELSD fingerprint of JOL was set up, and 15 batches of JOL were evaluated by similarity assay. Furthermore, the contents of the main 13 representative components were determined on the premise of small disparities among batches. Results: The HPLC-UVD-ELSD fingerprint of JOL was established with good separation, and 13 chemical components were determined simultaneously. Fifteen main characteristic peaks [gallic acid (peak 1), liquiritin (peak 5), aloe-emodin-8-O-β-D-glucopyranoside (peak 7), liquiritigenin (peak 11), baicalin (peak 13), chrysin-7-O-β-D-glucoronide (peak 16), oroxyloside (peak 17), wogonoside (peak 18), chrysophal-8-O-β-D-glucopyranoside (peak 19), chrysophal-1-O-β-D-glucopyranoside (peak 20), rhein (peak 24), glycyrrhizic acid (peak 26), (18β,20α)-glycyrrhizic acid (peak 27), hyodeoxycholic acid (peak 28), cholic acid (peak 29)] from four formula of JOL were chemically identified and 29 main characteristic peaks were assigned to individual herbs (peaks 8, 12, 13, 15-18 originate from Scutellariae Radix, peaks 3-5, 10, 11, 25-27 originate from Glycyrrhizae Radix et Rhizoma, peaks 1, 6, 7, 9, 14, 19, 20, 24 originate from Rhei Radix et Rhizoma, peak 2 originates from Fritillariae Ussuriensis Bulbus, peaks 28, 29 originate from Bovis Calculus Artifactus, peaks 21-23 originate from auxiliary materials). The similarity of 15 batches of JOL was about 0.968 to 1.000. Moreover, good linear relationships were found (R2=0.999 0-0.999 9), and the average recovery rates were 96.90%-102.84%. The content range of quantitative components in 15 batches of JOL (gallic acid 51.82-148.27 μg/mL, liquiritin 75.04-130.00 μg/mL, aloe-emodin-8-O-β-D-glucopyranoside 31.72-39.84 μg/mL, liquiritigenin 14.24-43.65 μg/mL, baicalin 610.37-867.40 μg/mL, chrysin-7-O-β-D-glucoronide 12.87-34.09 μg/mL, oroxyloside 62.45-101.48 μg/mL, wogonoside 155.41-205.86 μg/mL, chrysophal-1-O-β-D-glucopyranoside 11.56-23.72 μg/mL, chrysophal-8-O-β-D- glucopyranoside 16.14-36.87 μg/mL, glycyrrhizic acid 222.97-310.32 μg/mL, hyodeoxycholic acid 177.48-239.70 μg/mL, cholic acid 98.54-132.85 μg/mL) was determinated. Conclusion: The qualitative and quantitative methods of HPLC-UVD-ELSD mentioned above provided important evidence for further improving the overall quality standard of JOL.

Whole-ingredient identification and primary grade evaluation of Niuhuang Qingwei Pills based on reference drug / 中草药

Objective To identify whole ingredients in Niuhuang Qingwei Pills (NQP) and give primary grade evaluation of NQP. Methods Firstly, microscopic characteristics specified by the statutory standard of NQP were summarized. Then new microscopic identification methods were established for Sennae Folium and Gypsum Fibrosum while new thin layer chromatography (TLC) identification methods were established for Bovis Calculus Artifactus, Borneolum Syntheticum, and Rhei Radix et Rhizoma. In addition, microscopic identification method for Menthae Haplocalycis Herba was improved. In this way, whole-ingredient identification of 17 materials were realized. NQP reference drug was developed and applied as accompanying control in whole-ingredient identification and primary grade evaluation of the samples. Results According to the results of 48 samples from 18 manufactures by primary grade evaluation, 10, 37, and 1 samples were classified as superior, qualified, and unqualified, respectively. Conclusion The proposed methods are accurate, simple, and objective, which offers a more comprehensive approach for quality control of NQP. And the methods provide research strategy and experimental basis for further work on establishment of grade standard.

1H-NMR-based metabonomics study on toxicity alleviation effect of Bovis Calculus Artificialis, Gypsum Fibrosum, and Borneolum Synthcticum to Realgar (As2S2) in Niuhuang Jiedu Tablet / 中草药

Objective: To study the toxicity alleviation effect of Bovis Calculus Artificialis, Gypsum Fibrosum, and Borneolum Synthcticum to Realgar in Niuhuang Jiedu Tablet based on 1H-NMR metabonomics. Methods: Forty-two male Wistar rats were divided into six groups: group A (control group), group B (treated with realgar), group C (treated with Niuhuang Jiedu Tablet), group D (treated with Niuhuang Jiedu Tablet without Bovis Calculus Artificialis), group E (treated with Niuhuang Jiedu Tablet without Gypsum Fibrosum), and group F (treated with Niuhuang Jiedu Tablet without Borneolum Synthcticum). Based on 1H-NMR spectra of urine and serum from rats, PLS-DA was performed to identify different metabolic profiles. Results: The metabolic profiles of group B were different from control group, while the metabolic profiles of all other groups were almost in line with those of control group. Conclusion: The toxicity alleviation effects of Bovis Calculus Artificialis, Gypsum Fibrosum, and Borneolum Synthcticum to Realgar in Niuhuang Jiedu Tablet are not obvious.

Optimization of the Formulation and Technology of Compound Bovis Calculus Sativus Gel by Orthogonal Test / 中国药房

OBJECTIVE:To optimize the preparation technology of Compound bovis calculus sativus gel. METHODS:The ul-trasonic emulsifying technology was optimized by orthogonal test using ultrasonic power,ratio of ultrasonic time to interval time, total ultrasonic time as factors,using centrifugal stability constant(KE)as index.Ultrasonic emulsifying method was applied to pre-pare O/W emulsions using paeonol,berberine hydrochloride and eucalyptus oil;then calculus bovis sativus powder was added into O/W emulsions,and then mixed with carbomer(940)gel matrix to prepare gel. The formulation of gel was optimized by orthogo-nal test with the amount of carbomer (940),glycerool and triethanolamine as factors,using compactibility score,comprehensive score of release rate in vitro as index. Validation test,stability test and content determination of bilirubin were conducted for gel pre-pared by optimized technology. RESULTS:The optimal ultrasonic emulsifying technology was as follows as ultrasonic power 450 W,ratio of ultrasonic time to interval time 2:1,and total ultrasonic time 5 min. The optimal formulation of gel was as follows as carbomer(940)0.5%,glycerool 15%,triethanolamine 0.20%(g/100 g). The average of KE of validation test and average compre-hensive score were 0.175 and 98.67(RSD<2%,n=3);the appearance of the preparation had no obvious change in stability test, and average percentage of bilirubin in labeled content was 100.8%. CONCLUSIONS:The optimal formulation and preparation tech-nology of gel is feasible,and the prepared gel is stable and controllable in quality.

A rapid and effective hplc method for identification of bovis calculus in Chinese patent medicines / 中草药·英文版

Objective: Due to the limited resource and the large demand, many kinds of Bovis Calculus (BC) including artificial Bovis Calculus (ABC), in vivo cultured Bovis Calculus (in vivo CBC), and in vitro cultured Bovis Calculus (in vitro CBC) were used in Chinese patent medicines (CPMs). Previous studies have shown that the chemical constituents of ABC and their properties were different from other BC. The two types of CBC with much higher price than ABC were approximately equivalent with natural Bovis Calculus in quality and clinical effect. The aim of the study is to establish a rapid and effective method for the identification of BC in CPMs. Methods: An HPLC method with the higher specificity for analyzing bilirubin was established to distinguish ABC from other three kinds of BC by comparing the change of bilirubin content with the addition of EDTA-2Na as the extraction solvent and stabilizer. Results: The bilirubin content in CPMs containing ABC was basically unchanged, while that in CPMs containing other kinds of BC showed significant difference. The proposed method was employed to analyze a variety of CPMs containing Bovis Calculus (CPMBCs) and proven to be universal. Conclusion: An effective analytical method is established for the quality control of CPMBCs and further ensures the safety and efficacy of these drugs in clinical practice. © 2013 Tianjin Press of Chinese Herbal Medicines.