The differences and the variations of chondroitin sulfate content in different parts of Cervi Cornu Pantotrichum(CCP) with different processing methods were investigated. The chondroitin sulfate from velvet was extracted by dilute alkali-concentrated salt method. Next， the chondroitin sulfate was digested by chondroitinase ABC.The contents of total chondroitin sulfate and chondroitin sulfate A， B and C in the samples were determined by high performance liquid chromatography(HPLC).The content of chondroitin sulfate in wax，powder，gauze，bone slices of CCP with freeze-drying processing is 14.13，11.99，1.74，0.32 g·kg⁻¹， respectively. The content of chondroitin sulfate in wax，powder，gauze，bone slices of CCP with boiling processing is 10.71，8.97，2.21，1.40 g·kg⁻¹， respectively. The content of chondroitin sulfate in wax，powder，gauze，bone slices of CCP without blood is 12.47，9.47，2.64，0.07 g·kg⁻¹， respectively. And the content of chondroitin sulfate in wax，powder，gauze，bone slices of CCP with blood is 8.22，4.39，0.87，0.28 g·kg⁻¹ respectively. The results indicated that the chondroitin sulfate content in different processing methods was significantly different.The content of chondroitin sulfate in CCP with freeze-drying is higher than that in CCP with boiling processing.The content of chondroitin sulfate in CCP without blood is higher than that in CCP with blood. The chondroitin sulfate content in differerent paris of the velvet with the same processing methods was arranged from high to low as： wax slices， powder， gauze slices， bone slices.
Animals ; Chondroitin Sulfates ; analysis ; Deer ; Horns ; chemistry

This paper explored the specific peptides from Bubali Cornu by ultra-performance liquid chromatography-tandem mass spectrometry and based on mathematics set theory. Following the profile analysis of peptides from Bubali Cornu, Bovis Grunniens Cornu, Caprae Hircus Cornu, and Suis Cornu by nano LC-LTQ-Obitrap-MS after digestion with trypsin, the relationship of peptide composition among different samples was analyzed using the mathematics set theory. The ones that existed only in the Bubali Cornu set rather than in any other set were considered as the specific peptides of Bubali Cornu. The further bioinformatic analysis revealed four specific peptides from Bubali Cornu, whose specificity was verified by UPLC-QQQ-MS. The results showed that these four peptides could be used for distinguishing Bubali Cornu from Caprae Hircus Cornu and Suis Cornu. This study has provided a rapid and simple method for seeking the specific peptides in animal medicines, which can be utilized for quality evaluation of animal medicines, thus making them authenticable and traceable.
Animals ; Chromatography, Liquid ; Cornus ; Horns/chemistry* ; Peptides/chemistry* ; Tandem Mass Spectrometry

Mechanical properties and biological evaluation of buffalo horn material were examined in this study. The effects of sampling position of buffalo horn on mechanical properties were investigated with uniaxial tension and micron indentation tests. Meanwhile, the variation of element contents in different parts of buffalo horn was determined with elemental analysis, and the microstructure of the horn was measured with scanning electron microscopy. In addition, biological evaluation of buffalo horn was studied with hemolytic test, erythrocyte morphology, platelet and erythrocyte count, and implantation into mouse. Results showed that the buffalo horn had good mechanical properties and mechanical characteristic values of it gradually increased along with the growth direction of the horn, which may be closely related to its microstructure and element content of C, N, and S in different parts of the buffalo horn. On the other hand, because the buffalo horn does not have toxicity, it therefore does not cause hemolysis of erythrocyte and has a good affinity with it. Buffalo horn has good histocompatibility but meanwhile it may induce the platelet adhesion and aggregation. Even so, it does not continue to rise to induce a large number of platelet to aggregate with resulting blood clotting. Therefore, the buffalo horn material has been proved to possess good blood compatibility according to the preliminary evaluation.
Animals ; Biocompatible Materials ; Biomechanical Phenomena ; Buffaloes ; Erythrocytes ; Horns ; chemistry ; ultrastructure ; Mice ; Microscopy, Electron, Scanning

This study is to analyze and identify the water soluble components of water buffalo horn (Bubali Cornu, WBH), and also establish a method for investigating these components. Shotgun proteomic analysis identified proteins in WBH aqueous extraction: keratin, collagen, desmoglein, etc. Ultrafiltration and LC-MS/MS were used to separate and identify the peptides in WBH aqueous extract, as a result, identified peptides were mainly derived from nonspecific degradation products of keratin and collagen, which including C-terminal peptides and non C-terminal peptides. Hypoxanthine, uridine, guanosine, and adenosine were identified by comparing with the standards. The strategy in present study could be used in analyzing water soluble components of animal horn derived TCM. It provides a reference for investigation of the material basis of animal horn derived TCM.
Animals ; Buffaloes ; Chromatography, Liquid ; Guanosine ; Horns ; chemistry ; Hypoxanthine ; Mass Spectrometry ; Medicine, Chinese Traditional ; Peptides ; Proteomics ; Tandem Mass Spectrometry ; Uridine

OBJECTIVETo study the optimum extraction factors of Buxueyangyan Mixture.

METHODThe orthogonal design was used to select the optimum extraction factors of Cervi Pantotrichum etc. and water extraction rate was used to evaluate the factors. The orthogonal desingn was used to study three factors including crude particle size, extraction time and amount of water. Lcariine content was used as analytical parameter.

RESULTThe optimum extraction factors for Cervi Pantotrichum etc. were: 3 a volume ofwith 10 times of water used for three extraction 2.5 h each time. The optimum extraction factors for others were: 1 cm or smaller the crude particle size, boiling 2 times: 2.5, 1.5 h respectively the amount of water used was 10. 8 times of the drug mixture respectively.

CONCLUSIONAccording the optimum extraction factors, the active substance can be extracted efficiently.

Animals ; Deer ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; Epimedium ; chemistry ; Flavonoids ; analysis ; Horns ; chemistry ; Materia Medica ; isolation & purification ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Technology, Pharmaceutical ; methods ; Time ; Water

OBJECTIVETo study the effect of eight specifications of Cervi Cornu Pantotrichum on the osteoporosis of ovariectomized rats and grade the eight different specifications of Cervi Cornu Pantotrichum.

METHODTotally 100 SD female rats were divided randomly into 10 groups, namely the normal group, the model group and eight Cervi Cornu Pantotrichum groups of different specifications. Their bilateral ovaries were excised to reproduce the osteoporosis model. Meanwhile, the rats were given the eight different specifications of Cervi Cornu Pantotrichum for consecutively 12 weeks. Subsequently, the effects of the different specifications of Cervi Cornu Pantotrichum on bone mineral density and serum biochemical indicators of rats were observed. A clustering analysis was made for the eight specifications of Cervi Cornu Pantotrichum, with the serum content of ALP, BMP-2 and BGP as influencing factors.

RESULTAfter 12 weeks, the eight different specifications of Cervi Cornu Pantotrichum could significantly improve ALP, BMP-2, BGP in serum and bone mineral density of ovariectomized rats. And the cluster analysis showed similar results to the quality classification of traditional commercial herbs Cervi Cornu Pantotrichum.

CONCLUSIONDifferent specifications of Cervi Cornu Pantotrichum can antagonize the osteoporosis of ovariectomized rats, and their effects are related to the quality of commercial herbs.

Animals ; Bone Density ; drug effects ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Deer ; Disease Models, Animal ; Female ; Horns ; chemistry ; Humans ; Osteoporosis, Postmenopausal ; drug therapy ; genetics ; metabolism ; physiopathology ; Ovariectomy ; Rats ; Rats, Sprague-Dawley

To study the protective effect of Danqi Piantan capsule ( DPC) and its antelope horn substitution (DPCAS) on the cerebral ischemia, in order to preliminary study the possibility of replacing antelope horn with artificial bezoar. In this study, the left middle cerebral artery occlusion (MCAO) was adopted. Totally 150 SD rats were randomly divided into 5 groups: the sham operation group, the model group, the Danqi Piantan capsule (DPC) group (0.246 g x kg(-1) x d(-1)), the Danqi Piantan capsule without antelope horn (DPCRA) group (0.246 g x kg(-1) x d(-1)), the Danqi Piantan capsule without antelope horn and with double artificial bezoar (DPCDB) group (0.246 g x kg(-1) x d(-1)). The MCAO model was prepared 1 h later after the administration on the 5th day. At 24 h after the operation, the inner canthus blood was collected to determine the serum superoxide dismutase (SOD) activity and the endothelin (ET) content. At 72 h after the operation, the cerebral infarct size and the cerebral index were determined by TTC-staining. The fluorescent quantitative PCR method was used to detect brain Bcl-2, Caspase-3, IL-1β, P-selectin, E-selectin, ICAM-1 mRNA expressions. The mmunohistochemical method was used to detect ICAM-1, IL-1β, TNF-α, IL-6 expressions in ischemic penumbra. According to the results, compared with the model group, DPCDB and DPC groups showed almost consistent results, indicating both of the two group can significantly improved cerebral infarction index and cerebral index (P < 0.05), increase the serum SOD activity (P < 0.05), decrease the serum ET level and Caspase-3 expression, IL-1β, P-selectin, E-selectin, ICAM-1 mRNA expressions in brain tissues (P < 0.05) and expressions of ICAM-1, IL-1,6, TNF-α, IL-6 positive cells in ischemic penumbra (P < 0.05) and increase the Bcl-2 expression (P < 0.05). The DPCRA group showed much lower impacts on indexes than DPCDB and DPC groups. This suggests that DPCDB and DPC reveal similar efficacies and antelope horn in Danqi Piantan capsule can be substitutes by artificial bezoar.
Animals ; Antelopes ; Bile ; chemistry ; Biological Factors ; administration & dosage ; chemical synthesis ; chemistry ; Brain ; drug effects ; metabolism ; Caspase 3 ; genetics ; metabolism ; Drug Compounding ; Horns ; chemistry ; Humans ; Infarction, Middle Cerebral Artery ; drug therapy ; genetics ; metabolism ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Interleukin-1beta ; genetics ; metabolism ; Male ; Medicine, Chinese Traditional ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood ; genetics ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

The in vitro cell culture experiment was conducted to study the effect of Danqi Piantan capsule (DPC) and DPC dislodge the antelope horn with artificial bezoar double (DPCBD) on nerve regeneration and blood vessel regeneration and preliminarily investigate the possibility of substituting antelope horn in DPC with artificial bezoar. In this experiment, rats were randomly divided into 5 groups: the blank serum control group, the model group, DPC groups (0.306 g x kg(-1) x d(-1), the same below), DPC remove of antelope horn (DPCRA) groups and DPCBD groups. Brain microvascular endothelial cells cultured in vitro (BMEC), astrocytes and neural stem cells (NSC) were co-cultured to simulate neurovascular unit, label neurons with microtubule associated protein III (β-tubulin III) antibody and lable astrocytes with glial fibrillary acidicprotein (GFAP). ELISA was used for the detection of the content of BMEC lactate dehydrogenase instrument method (LDH), the inverted phase contrastmicroscope was adopted to observe the formation of BMEC tube like structure, the number of leukocytes and leukocytes adherent to BMEC were counted under the microscope, the expression levels of β-tubulin III and the ratio of GFAP positive cells was detected with inimmunofluorescence, and RT-PCR method was used to detect NGF, BDNF, VEGF and VEGFr-2 mRNA. According to the result, compared with the model group, both DPC and DPCBD can reduce LDH leakage, promote the formation of BMEC tube like structure, inhibit leukocytes and their adhesion to BMEC, increase the β-tubulin III positive cell differentiation proportion (P < 0. 01), reduce the proportion of GFAP positive cells (P < 0.01), increase the expressions of co-cultured NGF, VEGF, BDNF and VEGFr-2 mRNA to a certain extent, with the most significant difference on NGF and VEGF mRNA expressions (P < 0.05) and the same efficacy in both groups. DPCRA groups showed less impact on all indexes than that of DPCBD and DPC groups. The same efficacy of DPCBD and DPC on nerve regeneration and angiogenesis suggested that antelope horn in DPC can be substituted by artificial bezoar.
Animals ; Antelopes ; Brain-Derived Neurotrophic Factor ; genetics ; metabolism ; Cattle ; Cells, Cultured ; Drug Substitution ; Endothelial Cells ; drug effects ; metabolism ; Gallstones ; chemistry ; Horns ; chemistry ; Male ; Medicine, Chinese Traditional ; Nerve Growth Factor ; genetics ; metabolism ; Neural Stem Cells ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism