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Objective:To investigate the complex Calculus Bovis-target-keratitis network and to explore the molecular mechanism of Calculus Bovis treating keratitis through network pharmacology. Methods:Genes related to keratitis were searched in the online DisGeNET database and the protein-protein interaction (PPI) network of keratitis-associated proteins was constructed.The components isolated and identified in Calculus Bovis were collected through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP, https: //tcmsp-e.com/tcmsp.php), Chemistry Database by Shanghai Institute of Organic Chemistry of CAS (http: //www.organchem.csdb.cn), and published literature.The canonical SMILES information of the collected components was exported, which were submitted to the SwissTargetPrediction platform to predict potential targets of the components.The active component-predicted target network of Calculus Bovis was constructed and merged with the PPI network of keratitis-associated proteins to build the active component-potential target network of Calculus Bovis and systemically investigate the potential targets and signal pathways of Calculus Bovis in treatment of keratitis.The component-target-pathway network was established to analyze the mechanism of Calculus Bovis treating keratitis. Results:Thirty-nine components isolated and identified in Calculus Bovis were searched and 65 target genes related to keratitis were screened.Of the 28 potential targets involved in Calculus Bovis treating keratitis, there were 7 direct targets, including tumor necrosis factor, caspase 1, Toll-like receptor 9, C-X-C motif chemokine ligand 8, interleukin-6, mitogen-activated protein kinase 8, neurotrophic receptor tyrosine kinase 1.The 28 potential targets were annotated to 12 entries for biological process, 18 for cellular components and 13 for molecular function.In the Kyoto encyclopedia of genes and genomes pathway enrichment analysis, 10 signal pathways were identified as enriched categories, which were mainly related to human cytomegalovirus infection, amoebiasis, antifolate resistance, PI3K-Akt signaling pathway, rheumatoid arthritis, apoptosis, cytokine-cytokine receptor interaction, malaria, non-alcoholic fatty liver disease, interleukin-17 signaling pathway. Conclusions:Calculus Bovis may play an adjuvant therapeutic effect on keratitis through anti-inflammatory, antibacterial, antiviral, immune regulation, inflammatory regulation and other functions.
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Calculus Bovis is one precious traditional Chinese medicine used widely in clinical practice. Because of its high price and lack of resources, the substitutes of Calculus Bovis artificial Calculus Bovis, in vivo cultured Calculus Bovis and in vitro cultured Calculus Bovis also have been widely used in clinical practice. In order to understand the difference in chemical composition between Calculus Bovis and its substitutes, and determine whether all kinds of Calculus Bovis medicinal materials have special detection METHODS:, the chemical constituents and quality control METHODS: of Calculus Bovis and its substitutes were reviewed in this paper. Through the review of literature it was found that artificial Calculus Bovis mainly contain hyodeoxycholic acid and only a small amount of free bilirubin, and the chemical components of the other three kinds of Calculus Bovis medicinal materials are different. The chemical compositions of natural Calculus Bovis and in vitro cultured Calculus Bovis are similar, but the proportions of different types of bilirubin are different. At present, no characteristic ingredients of natural Calculus Bovis have been found to distinguish it from in vitro cultured Calculus Bovis and in vivo cultured Calculus Bovis.
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Calculus Bovis is the dry gall-stone of Bos taurus domesticus Gmelin, which is the most precious traditional Chinese medicine for its incomparable therapeutic effects. In vitro cultured Calculus Bovis,cultured Calculus Bovis and artificial Calculus Bovis are developed as the substitutes of Calculus Bovis. The Chinese Pharmacopoeia 2015 edition includes 89 Chinese patent drugs containing cow-bezoar and its substitutes. In this paper, the standards of Calculus Bovis and its substitutes in 89 Chinese patent drugs were reviewed to provide suggestions and reference for improving the relative standards. The qualities of Calculus Bovis and its substitutes of several Chinese patent drugs were summarized and analyzed based on the quality research of these drugs in recent years to offer suggestions for the revision of quality standards of relevant drugs and regulatory policy.
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The aim of this paper was to investigate the molecular mechanism of Calculus Bovis Sativus( CBS) in alleviating lipid accumulation in vitro by serum pharmacology. The CBS-containing serum of mice was obtained by serum pharmacology method to evaluate its effect on the proliferation of LO2 hepatocytes. The lipid reducing effects of CBS-containing serum through Nrf2 was evaluated by fructose-induced LO2 hepatocyte steatosis model,nuclear factor erythroid 2 related factor 2( Nrf2) agonist oltipraz combined intervention,cell oil red O staining and intracellular triglyceride( TG) content. The effects of CBS-containing serum on lipid peroxidation and hepatocytes apoptosis were evaluated by reactive oxygen species( ROS) and apoptosis assay,respectively. Real-time quantitative polymerase chain reaction( PCR) was used to detect the relative expression of lipid synthesis-related genes and apoptosis-related genes.RESULTS:: showed that CBS drug-containing serum had no significant effect on LO2 hepatocyte proliferation. As compared with the model group,CBS-containing serum could effectively reduce the formation of lipid droplets in fructose-induced LO2 hepatocytes,significantly reduce intracellular TG and ROS levels,and significantly reduce hepatocyte apoptosis rate( P < 0. 05). As compared with the model group,carbohydrate responsive element binding protein( ChREBP),sterol regulatory element binding protein-1 c( SREBP-1 c),fatty acid synthase( FAS),acetyl-CoA carboxylase 1( ACC1),stearoyl-CoA desaturase 1( SCD1),Bax and caspase-3 mRNA levels were significantly reduced in CBS drug-containing serum treatment group( P<0. 05). All of the above effects could be reversed by oltipraz.In conclusion,CBS-containing serum can significantly inhibit the fructose-induced LO2 liver fat deposition,and the mechanism may be related to reducing intracellular ROS level through the Nrf2 pathway and improving intracellular peroxidation state to reduce apoptosis.
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Animals , Cattle , Mice , Apoptosis , Cells, Cultured , Fatty Liver , Fructose , Gallstones , Chemistry , Hepatocytes , Cell Biology , Metabolism , Lipid Metabolism , Lipid Peroxidation , Liver , Medicine, Chinese Traditional , Reactive Oxygen Species , Metabolism , Serum , Chemistry , Sterol Regulatory Element Binding Protein 1 , Metabolism , TriglyceridesABSTRACT
Objective To develop an HPLC-MS/MS method for the simultaneous determination of 26 bile acids in Calculus Bovis Sativus (CBS) and Calculus Bovis (CB). Methods Chromatographic separation was performed on a Symmetry C18 column (150 mm × 2.1 mm, 3.5 μm) with linear gradient elution of water (0.1% formic acid + 10 mmol/L ammonium acetate) and methanol (0.1% formic acid + 10 mmol/L ammonium acetate) at a flow rate of 0.15 mL/min, and the injection volume was 10 μL. Switching electrospray ion source polarity in negative mode. The ion spray voltage was set at - 4 500 V, the turbo spray temperature was maintained at 350 ℃, and multiple-reaction monitoring (MRM) was employed. Results The 26 bile aicds showed good linearity within the linear ranges (r ≥ 0.991 4). The average recoveries of the bile acids ranged from 98.2% to 102.3%. The precisions (RSD) for the investigated components were less than 0.95%. There are some differences in variety and chemical components between CBS and CB. Conclusion The proposed method is reliable and has high sensitivity and good specificity, which allows the determination of various bile acids in CB.
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Calculus bovis(CB) exerts potencial pharmacological effects through anti-oxidation, anti-inflammatory, and anti-hypoxic injury. We reviewed the published literature about CB and its substitutes in the treatment of central nervous system diseases, in order to provide a reference for the deeper application and study in the future.
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Objective To observe the effect of saline gauze tamponade of calculus bovis sativus on the level of IL-6 and IL-8 in the tissue of patients with perianal abscess after operation,and to explore its possible mechanism of action in promoting wound healing.Methods Patients with 84 cases of anal abscess surgery,were randomly divided into treatment group(n=42)and control group(n=42).Treatment group calculus bovis sativus research pressed into powder,take 0.10~0.15g evenly scattered in the saline gauze on the wound packing,the control group was treated with physiological saline gauze wound.After treatment,3,7 and 10d were treated with local granulation tissue about 0.2cm×0.2cm×0.1cm size,the IL-6 and IL-8 cytokines in local tissues were detected by enzyme-linked immunosorbent assay.Results There was no significant difference in the expression of IL-6 and IL-8 between the two groups 3 days after treatment(P>0.05),IL-6 and IL-8 in the treatment group(7,10d)were significantly lower than that in control group,and the difference was statistically significant(P<0.05).Conclusion Calculus bovis sativus in the treatment group,IL-6 and IL-8 levels were significantly lower than those of the control group,indicating that calculus bovis sativus can reduce local inflammation response,promote healing of wound after perianal abscess operation.
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Calculus Bovis (CB) is one of the precious traditional Chinese medicines (TCM) and has been widely used in oriental countries over 2000 years.It possesses a broad spectrum of pharmacological properties such as relieving fever,analgesic,diminishing inflammation,sedation,reducing blood pressure,recovering gallbladder functions,antioxygenation,and anti-tumor and improving immunity,et al.Due to the scarce resource and high price of natural CB,its substitutes,artificially synthesized CB and C.Bovis Sativus (CBS),also called in vitro cultured C.Bovis,have already been developed and widely used in medicine preparation.By using the literature material method,the current article explored literatures regard to pharmacological activities and clinical application of CB and its substitute,and the compound preparations were also included,from 1949 to 2016 in journals of domestic and foreign.All documents are summarized to provide a valuable reference for further basic or clinical research about CB and its substitute.
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Natural Calculus Bovis,one of the precious traditional Chinese medicines,is the dried gallstone in bovine gallbladder,bile duct or hepatic duct.It is one of the essential components of the formulation of a variety of traditional Chinese medicines.Due to high price and rareness of natural Calculus Bovis,Chinese researchers have developed artificial Calculus Bovis,cultured Calculus Bovis and Calculus Bovis Sativus as substitutes of it.They are mainly composed of bilirubin,bile acids,amino acids and inorganic elements.Due to the co-existence of natural Calculus Bovisa and its substitutes in the market,and the complex composition of them,identification,quality control and material basis study of them are important issues in recent years.Currently,the quality control of them mainly focus on cholic acid and bilirubin.In order to fully solve the quality control problem of natural Calculus Bovis and its substitutes,researches on testing techniques of other bile acids or other components have already started.In this paper,research progress on the chemical compositions,quality control and the test technology of bilirubin,bile acids,amino acids and other components of Calculus Bovis are reviewed,in order to provide reference for further research of Calculus Bovis.
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The preparation of Calculus bovis and its compounding preparations have been used widely in clinical practice.Traditionally,the forms of medicine were in raw material medicine way,preparing tablet,pill,powder or directly taking its powder.The main active ingredient of Calculus bovi were considered to be bilirubin and bile acids.However,the traditional formulations caused low bioavailability and wasted expensive herbs because its main component were insoluble in water.In recent years,many researchers have tried to use modern preparation technology to prepare its compounding formulations,such as solid dispersion technology,ultrafine grinding technology,powder coating technology,liposome encapsulation technology,or simplifying the prescription by using of known pharmacological effects of soluble components as substitutes.These methods were considered to be feasible to develop new formulations of Calculus bovis.In this paper,in order to provide reference of method and technology for the improvement of Calculus bovis compounding preparation and the development of new dosage form,ultramicrostructure,chemical composition,improvement methods and techniques of compounding preparation were analyzed.In addition,the relevant techniques and method of improving the formulation Calculus bovis compounding preparation in recent years were reviewed.
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Calculus Bovis is a valuable traditional Chinese medicine and has been used for more than two thousand years in clinic with the effects of puring heart, sweeping phlegm, resuscitation, extinguishing wind and detoxification. Since the founding of the People's Republic of China, modern methods have been utilized by traditional Chinese medicine researchers in the resource identification, chemical components, pharmacodynamics, pharmacokinetics, pharmacy, clinical application, etc. It is their continually exploration that makes significant achievements for the modern research of Calculus Bovis. This article statistically analysed the literatures from 1949 to December 2015 in Pubmed, CNKI, Wanfang, Vip database etc. to review Calculus Bovis and its compound formulas, as well as its substitutes, quality control, formulation study, compound prescription, pharmacological mechanism and clinical research. The aim of this article is to provide a valuable reference for future developments and studies of Calculus Bovis.
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OBJECTIVE: To develop a method for determination of different types of bilirubin in bezoar and explore the differences in their contents or ratios between natural and in vitro cultured Calculus Bovis to provide reference for identification of Chinese patent drugs containing bezoar. METHODS: Free bilirubin was determined directly by HPLC, while unconjugated and total bilirubin in bezoar were converted into free bilirubin respectively using different chelating agents. Converted bilirubin was eluted on a Diamonsil C18 column(4.6 mm×150 mm, 5 μm)using isocratic mobile phase composed of acetonitrile-1% acetic acid (95∶5) and detected at 450 nm. RESULTS: A small amount of free bilirubin was detected in Calculus Bovis but not found in in-vitro cultured Calculus Bovis. The content of total bilirubin in Calculus Bovis from different sources varied greatly while being similar in in vitrocultured samples. Bilirubin in covalent form occupied a great part in Calculus Bovis, and bilirubin in conjugated form hold a great proportionin in vitro cultured Calculus Bovis. CONCLUSION: The contents or ratios of different types of bilirubin can be used as the quality control parameters of naturaland in vitro cultured Calculus Bovis. The established method is easy to operate with high effectiveness in quality control, suggesting a new way of identifying bezoar.
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Nature Calculus Bovis is one of the precious traditional Chinese medicines, which is the dried gallstone in gallbladder, bile duct or hepatic duct. In order to meet the clinical needs, we have developed artificial Calculus Bovis, invitro and other varieties. Calculus Bovis mainly contains bile pigments, bile acids, amino acids, inorganic elements and other types of components. Besides bile pigments, bile acids is the second largest of the active ingredients of Calculus Bovis, which is closely related to the antipyretic and analgesic, antiinflammatory effect and depressurization. The article outlines and summarizes the study progress in pharmacological effects and quantitative methods for components of bile acids in Calculus Bovis and its substitutes and preparations, aiming to provide a reference for subsequent research as well as quality control.
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Objective To establish a pre-column derivation HPLC-UV method for the content determination of hyodeoxycholic acid in artificial calculus bovis. Methods The hyodeoxycholic acid was derived by 2-bromo-2’-acetonaphthoneat using triethylamine as the catalyst in 60 ℃ water bath. After that, a HPLC-UV method was established to determine the content of hyodeoxycholic acid in artificial calculus bovis. Results When the derivatising time at 60 ℃ water bath was 50 min, the radio of the molar amount of derived reagents and hyodeoxycholic was over 20:1 and the radio of catalyst and hyodeoxycholic was over 15:1; the reaction was completed. The calibration curve was linear within the range of 0.1–2 μg for hyodeoxycholic acid (r=0.999 7), and the average recovery was 97.85% (RSD=1.6%). In this sample, the content of hyodeoxycholic is 4.12%. Conclusion The method is with high sensitivity, highly reproducible, reliable and accurate for the content determination of hyodeoxycholic acid in artificial calculus bovis.
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OBJECTIVE: To study the type and quality of Calculus Bovis in commercial Angong Niuhuang Pills (traditional Chinese medicines) and provide technical support to fight against counterfeits. METHODS: Specific and effective components of Calculus Bovis were studied. The type of Calculus Bovis in commercial Angong Niuhuang Pills was identified according to the differences in components and content. RESULTS: Illegal use of artificial Calculus Bovis or bilirubin was found to be a serious problem. The type and quality of Calculus Bovis in Angong Niuhuang Pills differed greatly. CONCLUSION: Calculus Bovis adulteration in Angong Niuhuang Pills is a very serious problem which needs urgent supervision and regulation to assure safe medication.
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The protective effects of in vitro cultivated calculus bovis (ICCB) on the cerebral and myocardial cells in hypoxic mice and the mechanism were examined. In one group, mice were intra-gastrically (i.g.) given ICCB for 15 days and then they were subjected to acute cerebral ischemia by decapitation, and then the panting time was recorded. In the other group, 12 min after exposure to hypoxia, mice was administered the ICCB i.g. for 5 days, and then the blood serum and tissues of brain,heart, liver were harvested and examined for SOD, GSH-px and T-AOC activity and content of MDA. The tissues of brain and heart were observed electron-microscopically for ultrastructural changes. The corpus striatum and hippocampus of brain were collected and examined for content of dopamine (DA) and norepinephrine (NE). The ultrastrural examination showed that the pathological change in brain and heart in the ICCB group was very slight, while abnormal changes in the control group were obviously more serious. ICCB significantly prolonged the panting time of the hypoxic mice (P<0.001), increased the activity of SOD, GSH-px, T-AOC in serum and tissues of brain, liver,heart and elevated the content of DA and NE. ICCB also pronouncedly reduced content of MDA in serum and tissues of brain, heart and liver. Significant differences in these parameters were noted between ICCB group and controls. It is concluded that ICCB can exert protective effect on the cells of brain and myocardium by enhancing the tolerance of the tissues to hypoxia and the body's ability to remove free radicals and regulating the neurotransmitters.
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AIM: To probe into the anti-epilepsy action of artificial Calculus Bovis,by observing its effect on the behavioral of the experimental epileptic rats,neuron loss in the hippocampus and hilus,and GAD positive cell alteration in the hippocampus.METHODS: SD rats were divided into three groups: group A(artificial Calculus Bovis treatment group);group B(acute epilepsy group) and group C(control group).A model of acute epilepsy rats was established by PTZ.The rat's behavioral alteration was observed by the Racine' scale.The neurons in the hippocampus and hilus were calculated by Nissl staining.The GAD positive cells were observed by immunohistochemical staining.RESULTS: The latency of the first seizure in group A was longer than that in group B,while the seizure times in group A was less than that in group B.Besides,in group A,both the neuron loss amount in the hippocampus and hilus and the GAD positive cell loss amount in the hippocampus were less than those in group B.CONCLUSION: The artificial Calculus Bovis prolonged the latency of the first seizure time,decreased the frequency of seizure,and prevented the neuron loss and protected the GAD positive cells.
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Objective To make a qualitative and a quantitative control on the Calculus Bovis in Houzao Niuhuang Powder.Methods Hyodeoxycholic acid and bilirubin in Calculus Bovis were separately identified by TLC,and the cholic acid content was determined by TLCS with the scan wavelength of ?s=380 nm,?R=650 nm.Results TLC can be used for the identification in-vitro cultured Calculus Bovis and artificial Calulus Bovis,the samples containing cultured Calulus Bovis showed the specific spots of bilirubin but no hyodeoxycholic acid spot while the samples containing artifical Calulus Bovis showed the hyodeoxycholic acid spot but no bilirubin spot.TLC-scanning can be used to measure the quantity of cholic acid,with the linear range being 0.2～1.8 ?g,average recovery rate of 99.5 %,and RSD being 3.3 %(n=6),and the results accorded with the criterion of the cultured Calculus Bovis in Chinese pharmacopoeia.Conclusion The methods can effectively control the quality of Calculus Bovis in Houzao Niuhuang Powder.
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0.05).The results of electrocardiography and the laboratory ex-amination showed that neither ANWin cluding natural Calculus Bovis nor A NWincluding in -vitro cultured Calc ulus Bo-vis had obviously toxic and side effe cts in treating epidemic encephalitis B.Conclusion ANW including in -vitro cul-tured Calculus Bovis has an markedly effect in the treatment of epidemic e ncephalitis B.
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0.05).Conclusion In-vitro-cultured CB has good effects in the treatment of apoplexy.Neither in-vitro-cultrued CB nor natural CB for apoplexy has obvious adverse reaction.