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Objective To study the effect of velvet antler polypeptides (VAP) on Rho/ROCK pathway in APP/ PSl double transgenic mice. Methods APP/PSl double transgenic mice were randomly divided into model group and velvet antler polypeptide group, 20 mice in each group, and control group consisting of 20 mice of the same litter and the same gender negative. The mice in VAP group were given velvet antler polypeptide 100 mg/kg by intragastric administration once a day for 28 days. After treatment, the water maze experiment was detected and recorded the escape latency and the number of crossing platforms of the mice; the ultrastructures of the synapse were observed by transmission electron microscopy; the expression of Rhs homolog gene family member A(RhoA) and Rho associated coiled-coil forming protein kinase II(ROCKII) in the hippocampal CAI area were observed by immunofluorescence. The expression levels of RhoA and ROCKII protein in the hippocampus were detected by Western blotting. The contents of hippocampus amyloid (3-protein(A(3),
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Objective To study the effect of velvet antler polypeptides (VAP) on antioxidant in Alzheimer' s disease model mice. Methods Eight months old male amyloid precursor protein (APP)/presenilin-l (PS1) double transgenic mice were selected as Alzheimer' s disease (AD) model and divided into the model group and the VAP intervention group, 12 in each group. Besides, normal mice of the same brood (with no transgene) were recruited as a control group (n= 12).After 6 months of intragastric administration, behavior, morphology and oxidative stress related indicators were detected.SH-SY5 cells were used to establish AD model of damaged by Ap2535. The expression levels of APP and p-secreatase-l(BACE1) protein in mouse hippocampus were detected by Western blotting. VAP intervention group SH-SY5Y cells was cultured with VAP (500 g/L) and amyloid P(Ap) 2535(25 ixmol/L) for 24 hours. Control group cells were normally cultured by DMEM medium. Cell apoptosis, membrane potential, reactive oxygen species (ROS) levels and oxidative stress related indexes were detected. Results In animal models, compared with the model group, the escape latency of mice in the VAP intervention group was shortened (P<0. 05). The neuronal cells in the CA1 region of the hippocampus of the model group were reduced and arranged disorderly. The arrangement of the VAP intervention group was relatively regular, and the morphology was significantly improved. Compared with the model group, senile plaques were decreased in the VAP intervention group. Compared with the model group, the malondialdehyde (MDA) content ol the VAP intervention group increased, and the superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) content increased, the difference was statistically significant. Compared with the control group, the APP and BACE1 content in the model group increased. Compared with the model group, the contents of APP and BACE1 in the VAP intervention group decreased, and the difference was statistically significant (P<0. 05). In the cell model, the apoptosis rates of the VAP intervention group decreased. Compared with the model group, the mitochondrial membrane potential of the VAP intervention group increased, the content ol ROS decreased, the content of MDA decreased, and the content of SOD and GSH-Px increased. The difference were statistically significant (P<0. 05). Conclusion VAP has a protective effect on oxidative stress damage caused by Alzheimer' s disease model animals and cells, which may be achieved by reducing ROS production and increasing the activity of antioxidant enzymes to reduce Ap deposition.
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Objective @#To explore the feasibility of antler powder/silk fibroin/polyvinyl alcohol scaffolds as tissue engineering bone scaffolds and the relationship between their degradation performance and the healing speed of bone defects.@*Methods @# Antler powder/silk fibroin/polyvinyl alcohol scaffolds and nano hydroxyapatite/silk fibroin/polyvinyl alcohol scaffolds were prepared by 3D printing. The whole bone marrow culture method was used to prepare blood cell sheets of Altay big tail sheep’s iliac bone marrow. With observation times of 1, 2 and 3 months, the mandibular defects of 4 sheep were established. The experimental group was coated with antler powder/silk fibroin/polyvinyl alcohol scaffolds. The control group was coated with nanohydroxyapatite/silk fibroin/polyvinyl alcohol scaffolds. The negative control group was coated with gel-free sponges. According to the self-control method of the bilateral mandible defect area, scaffolds wrapped with cell membranes or gel sponges wrapped with cell membranes were implanted. At the ends of the first, second and third months after implantation, the experimental animals were killed, cone beam CT was performed, and paraffin sections were taken for HE staining to evaluate the effect of different scaffold materials on bone regeneration and scaffold degradation.@* Results@# Scanning electron microscopy showed that both groups had regular pores and good continuity, and there was no difference in pore size and porosity between the two groups (P > 0.05). The results of CBCT imaging showed that in 3 months after operation, the experimental group had significantly better repair effects on bone defects than the control group, and the degradation rate matched the bone repair rate. The bone mineral density in the center of the defect was higher than that of the control group, which was close to that of normal bone tissue. The central bone mineral density of the experimental group at each time point was higher than those of the control group and the negative control group, and the difference was statistically significant (P < 0.05). HE staining results showed that the bone cells in the experimental group were more active, with more new capillaries and bone trabeculae formed, and the scaffold material absorbed more than the control group. @*Conclusion @#The antler powder/silk fibroin/polyvinyl alcohol scaffold can promote the repair of critical bone defects. Its degradability matches its bone tissue healing rate. It is expected to become a promising scaffold material for bone tissue engineering.
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Objective:To investigate the intervening effect of velvet antler peptide （VAP） on rotenone-induced neuroblastoma （SH-SY5Y） cell damage and explore its related mechanism. Method:0.5 μmol·L-1 rotenone was used to SH-SY5Y cells to establish an in vitro model of Parkinson's disease （PD）. A blank control group， a model group， high， medium and low dose VAP groups （150，100，50 mg·L-1， respectively） and a rapamycin group were established. The number of lewy bodies， changes in mitochondrial membrane potential， content of reactive oxygen species （ROS） and α-synuclein （α-syn）， protein kinase B （Akt）， and mammalian target of rapamycin （mTOR） were observed by hematoxylin-eosin（HE） staining， rhodamine 123 staining， DCFH-DA staining and immunohistochemical staining expression respectively. Result:The results of HE staining showed that as compared with the blank group， the number of cells in model group was reduced， the tentacle structure became dull， the shape became round， and eosinophilic Lewy bodies were visible in cytoplasm. As compared with model group， there was no significant difference in cell morphology from rapamycin group and VAP high， medium and low dose groups， but there were fewer Lewy bodies in cytoplasm in these four groups. Rhodamine 123 staining showed that as compared with blank group， the mitochondrial membrane potential was increased significantly in model group （P<0.05）. As compared with the model group， the mitochondrial membrane potential was decreased in rapamycin group and VAP high， medium and low dose groups （P<0.05）. DCFH-DA staining results showed that as compared with blank group， the content of ROS was increased significantly in cells of model group （P<0.05）. As compared with model group， the content of ROS was decreased in rapamycin group and VAP high， medium and low dose groups （P<0.05）. Immunohistochemical staining showed that as compared with blank group， the protein expression levels of α-syn，Akt，and mTOR were increased significantly in model group （P<0.05）. As compared with model group， the protein expression levels of α-syn and mTOR were significantly reduced in rapamycin group and VAP high and medium dose groups （P<0.05）， and the expression levels of Akt were significantly reduced in rapamycin group and VAP high-dose group （P<0.05）. Conclusion:Velvet antler peptides may play a neuroprotective role by regulating the Akt/mTOR signaling pathway and promoting the degradation of α-syn in SH-SY5Y cells.
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OBJECTIVE: To study the DNA fingerprint characteristics of the precious Chinese medicinal antler, identify and analyze the velvet antler at the molecular level, and develop the velvet DNA detection kit and evaluate its performance. METHODS: Using antler cytochrome C oxidase subunit Ⅰ (Co Ⅰ) and velvet mitochondrial cytochrome b (cytochrome b, Cyt b) as target genes, five pairs of specific primers were designed by bioinformatics technology, and the oxidase subunit Ⅰ of cytochrome C, mt DNA CoⅠ 1 (NC_013834.1), was determined to be the best specific primer by experimental screening. Polymerase chain reaction (PCR) amplification was conducted on velvet antler samples, and the PCR reaction system and reaction conditions were optimized to develop a velvet DNA detection kit. The performance of the kit was evaluated and commercially available antler samples were tested. RESULTS: The purity of the DNA extracted by the antler DNA test kit was 1.785-1.906. The specificity of the kit was reflected in the detection of genuine bands, and there was no band in the fakes. The sensitivity was up to 3.125 ng•μL-1. The kit was still effective after 5, 10, 15 and 20 freeze-thaw cycles. The results of repeated detection of positive and negative products for three times were consistent. The kit could be stored for up to one year at -20 ℃. The quality analysis was carried out on 12 commercial samples in five regions, and the pass rate was 66.67%. CONCLUSION: The velvet DNA detection kit developed by our team has strong specificity, high sensitivity, good stability and repeatability, with simple operation, small sample volume, wide application range, low cost and accurate results. It provides a scientific, accurate and reliable identification method for Chinese medicinal materials velvet products. At the same time, it also shows that the quality of commercially available velvet samples is uneven, and there are many adulterants.
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BACKGROUND: The proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) can delay the procession of steroid-induced femoral head necrosis. Besides, microRNA-141 (miR-141) is one of the important regulatory factors to promote cell proliferation. In addition, velvet antler is a traditional Chinese medicine which has significant roles in repairing bone and tissue and improving health. OBJECTIVE: To investigate whether velvet antler serum can regulate the expression of miR-141 to promote the proliferation of BMSCs, and further delay or reverse the progression of steroid-induced femoral head necrosis. METHODS: BMSCs were isolated and cultured from Sprague-Dawley rats. The passage 3 BMSCs were transfected with miR-141 mimic or miR-141 inhibitors, and then real-time PCR and methyl thiazolyl tetrazolium (MTT) assay were performed for detecting miR-141 expression and cell proliferation, respectively. The passage 3 BMSCs were divided into three groups: control group (α-MEM), dexamethasone group (α-MEM+1 μmol/L dexamethasone), and velvet antler serum group (α-MEM+1 μmol/L dexamethasone+15% velvet antler serum). Expression of miR-141 mRNA was detected by real-time PCR at 24 hours after intervention. The proliferation ability of BMSCs was evaluated by MTT assay at 24, 48, and 72 hours after intervention. RESULTS AND CONCLUSION: After transfection with miR-141 mimic, the expression of miR-141 mRNA was upregulated, while the cell proliferation was reduced. After transfection with miR-141 inhibitor, the expression of miR-141 mRNA was downregulated, while the cell proliferation was increased. The expression of miR-141 mRNA was significantly higher in the dexamethasone group than the control group (P < 0.01), while the treatment with velvet antler serum could significantly downregulate the expression of miR-141 mRNA (P < 0.01). The absorbance of BMSCs in the dexamethasone group was significantly lower than that in the control group (P < 0.01), and the absorbance value in the velvet antler serum group was significantly higher than that in the dexamethasone group (P < 0.01). In conclusion, the serum containing velvet antler can downregulate the expression of miR-141 which is upregulated by dexamethasone and then do help to promote the proliferation of BMSCs.
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Antler growth is a unique event compared to other growth and development processes in mammals. Antlers grow extremelyfast during the rapid growth stage when growth rate peaks at 2 cm per day. Antler growth is driven by a specificendochondral ossification process in the growth center that is in the distal region of the antler tip. In this study, we usedstate-of-art RNA-seq technology to analyze the expression profiles of mRNAs and miRNAs during antler growth. Ourresults indicated that the expression levels of multiple genes involved in chondrogenesis and endochondral ossification,including Fn1, Sox9, Col2a1, Acan, Col9a1, Col11a1, Hapln1, Wwp2, Fgfr3, Comp, Sp7 and Ihh, were significantlyincreased at the rapid growth stage. Our results also indicated that there were multiple differentially expressed miRNAsinteracting with differentially expressed genes with opposite expression patterns. Furthermore, some of the miRNAs,including miR-3072-5p, miR-1600, miR-34-5p, miR-6889-5p and miR-6729-5p, simultaneously interacted with andcontrolled multiple genes involved in the process of chondrogenesis and endochondral ossification. Therefore, we established a miRNA-mRNA regulatory network by identifying miRNAs and their target genes that were differentially expressedin the antler growth centers by comparing the rapid growth stage and the initial growth stage
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OBJECTIVE: To investigate the color and chemical composition of the wax slices, w-powder slices, r-powder slices, blood slices and bone slices of two-branch Velvet Antler, three-branch Velvet Antler and reborn Velvet Antler with different growth years, and analyze the relationship between the color characteristics and chemical composition (protein, polysaccharide, phospholipid). METHODS: The color parameters of five kinds of slices with different growth periods were detected by CIEL*a*b* color space. The content of protein was determined by Coomassie blue colorimetric method, the content of polysaccharide was determined by phenol sulfuric acid method, and the content of phospholipid was determined by molybdenum blue colorimetric method. Pearson correlation analysis was used to analyze the correlation between color and components. RESULTS: There were significant differences in color and chemical composition between the five types of Velvet Antlers with different grow periods. Pearson correlation analysis showed that the Velvet Antler color parameter b* had a significant negative correlation with the three chemical components. CONCLUSION: The color determination method based on the principle of colorimetry can effectively distinguish Velvet Antlers in different growth stages. And the correlation analysis showed that the color digital index can reflect the difference in chemical composition of Velvet Antler in a certain degree.
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Objective:: To investigate the effects of pilose antler polypeptide on the abilities of proliferation and collagen secretion of mouse embryonic fibroblasts NIH/3T3, and to clarify the relevant mechanisms. Methods: The NIH/3T3 cells were treated with different doses 1.56, 3.13, 6.25, 12.50, 25.00, 50.00, 100.00, and 200.00 mg • L_ 1) of pilose antler polypeptide as experimental groups, the cells treated with 0 mg • L_ 1 pilose antler polypeptide were used as blank control group, and the cells treated with 50.00 fig • L-1 basic fibroblast growth factor (bFGF) were used as positive control group. MTT assay was used to detect the survival rates of NIH/3T3 cells in various groups. ELISA assay was used to detect the collagen secretion of NIH/3T3 cells in various groups. Wound healing assay was used to detect the migration abilities of NIH/3T3 cells. Western blotting method was performed to detect the expression levels of p-ERK 1/2 in the NIH/3T3 cells in various groups. Immunofluorescence method was used to detect the expression levels of transforming growth factor-fil (TGF-J31) in the NIH/3T3 cells in various groups. Results: Compared with blank control group, the survival rates of NIH/3T3 cells in positive control group and 6.25, 12.50, 25.00, 50.00, 100.00, 200.00 mg • L_ 1 pilose antler polypeptide groups were markedly increased (P < 0 . 05 or P < 0 . 01). Compared with blank control group, the levels of type I collagen protein in the culture solution of the NIH/3T3 cells in positive control group and 6. 25, 12. 50, 25. 00, and 50. 00 mg • L_ 1 pilose antler polypeptide groups were markedly increased (P < 0 . 05 or P < 0 . 01), and the levels of type IE collagen protein in the culture solution of the NIH/3T3 cells in positive control group and 12. 50 and 25.00 mg • L_ 1 pilose antler polypeptide groups were markedly increased (P < 0 . 05). Compared with blank control group, the scratch healing rates of NIH/3T3 cells, and the expression levels of p-ERK 1/2 in the NIH/3T3 cells, and the expression levels of TGF-J31 in the NIH/3T3 cells in positive control group and 12. 50 mg • L_ 1 pilose antler polypeptide groups were markedly increased (P < 0 . 05 or P < 0 . 01). Conclusion: Pilose antler polypeptide can promote the proliferation, and collagen secretion of NIH/3T3 cells and increase the migration ability, which may be achieved by activating the phosphorylation of ERK 1/2 and increasing the expression of TGF-J31.
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Eleven steroid hormones (SHs: androstene-3,17-dione, estrone, β-estradiol, α-estradiol, testosterone, dehydroepiandrosterone, 17á-hydroxyprogesterone, medroxyprogesterone, megestrol acetate, progesterone, and androsterone) were detected from New Zealand deer (Cervus elaphus var. scoticus) velvet antler (NZA, 鹿茸). A method for the quantification of eleven SHs was established by using ultraperformance liquid chromatography (UPLC)-MS/MS. The linearities (R² > 0.991), limits of quantification (LOQ values, 0.3 ng/mL to 23.1 ng/mL), intraday and interday precisions (relative standard deviation: RSD 0.999), LOQ values (30 ng/mL to 350 ng/mL), intraday and interday precisions (RSD < 1.93%), and recovery rates (97.2% to 103.5%) for the three 7-O-CSs were determined. These quantitative methods are accurate, precise, and reproducible. As a result, it is suggested that the five steroid compounds of androstene-3,17-dione, androsterone, 7-ketocholesterol, 7α-hydroxycholesterol, and 7β-hydroxycholesterol could be marker steroids of NZA. These methods can be applied to quantify or standardize the marker steroids present in NZA.
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Animals , Androsterone , Antlers , Chromatography, Liquid , Deer , Dehydroepiandrosterone , Estrone , Medroxyprogesterone , Megestrol Acetate , Methods , New Zealand , Progesterone , Steroids , TestosteroneABSTRACT
Objective:To explore the effect and mechanism of pilose antler different components on the bone tissue of ovariectomized osteoporosis model rats and ascertain the material basis of pilose antler. Method:fifty-six SD rats were divided randomly into seven groups:normal group,model group,Xianling Gubao group(468 mg·kg-1),Bujiale group(80 mg·kg-1),polysaccharide group(50 mg·kg-1),polypeptides group(175 mg·kg-1),polysaccharide and polypeptide mixture group(50 mg·kg-1+175 mg·kg-1). Osteoporosis mode was established through ovary resection of female rats,meanwhile,the rats were given different components of pilose antler for consecutively 12 weeks. Subsequently, using absorptiometry to measure the rats' bone mass density. The activities of bone alkaline phosphatase(BALP),osteocalcin (OT),bone morphogenetic protein2(BMP-2),Smad1,Smad5,Runt-related transcription factor 2 (RUNX2) were detected by enzyme-linked immuno sorbent assay (ELISA). The expression of BMP-2,Smad1,Smad5,Runx2 protein was examined by Western blot and Real-time polymerase chain reaction (Real-time PCR). Morphological assay for bone tissue were detected by htoxylin eosin(HE) staining. Result:After 12 weeks, Compared with the normal group, the osteoporosis model group showed significantly decrease in bone mineral density(PPPConclusion:Pilose antler different components has therapeutic effect on ovariectomized osteoporosis model rats.The mechanism may be related to up-regulat the expression of BMP-2/Smad1,Smad5/Runx2 signal pathways.
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Peptides from Pilose antler aqueous extract (PAAE) have been shown to stimulate the proliferation and differentiation of bone marrow mesenchymal stem cells (BMSCs). However, the underlying molecular mechanisms are not well understood. Here, PAAE was isolated and purified to explore the molecular mechanisms underlying PAAE's effects on BMSCs as well as its osteoprotective effects in ovariectomized rats. Our results showed that PAAE promoted proliferation and differentiation of BMSCs to become osteoblasts by enhancing ALP activity and increasing extracellular matrix mineralization. The trabecular microarchitecture of ovariectomized rats was also found to be protected by PAAE. Quantitative reverse transcription-polymerase chain reaction (Quantitative RT-PCR) results suggest that PAAE also increased the expression of osteogenic markers including, alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), osteocalcin (OCN), bone morphogenetic protein-2 (BMP-2), and collagen I (COL-I). Immunoblotting results indicated that PAAE upregulated the levels of BMP-2 and Runx2 and was associated with Smad1/5 phosphorylation. PAAE A at the concentration of 200 μg·mL showed the strongest effect on proliferation and osteogenic differentiation of BMSCs after 48 h. Using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS), we identified the molecular weight of PAAE A and found that it is less than 3000 Da and showed several significant peaks. In conclusion, PAAE activates the BMP-2/Smad1, 5/Runx2 pathway to induce osteoblastic differentiation and mineralization in BMSCs and can inhibit OVX-induced bone loss. These mechanisms are likely responsible for its therapeutic effect on postmenopausal osteoporosis.
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Differential proteomics analysis of Sika deer antlers at rapid growth stage (60 d) and ossification stage ( 90 d) was performed by isobaric tags for relative and absolute quantitation ( iTRAQ ) , ultra high performance liquid chromatography and mass spectrometry technologies. A total of 127 differential proteins were identified. Compared with the ossification stage, 80 differential proteins were significantly up-regulated and 47 differential proteins were significantly down-regulated at the rapid growth stage. These differential proteins were mainly distributed in the regions of extracellular matrix, nucleosome, haptoglobin-hemoglobin complex, actin filament, endoplasmic reticulum-Golgi intermediate compartment, endoplasmic reticulum lumen, and endometrium, etc. The up-regulated differential proteins were mainly involved in the regulations of oxygen transport in the blood, nerve growth and regeneration, cartilage and bone development and ATP synthesis compared with ossification stage, and the down-regulated differential proteins were mainly involved in the endochondral ossification process. The changes of protein expression at different growth stages were closely related to antler rapid growth and ossification. Therefore, the results of this study provided a basic data for discovering the molecular mechanisms of antler rapid growth and ossification, and it was of great significance for further study of the pharmacological basis and clinical application of antlers.
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Reversed phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS) was utilized to investigate peptide profiling and bioactivities of antler aqueous extract(AAE),digested antler aqueous extract (AED) and powder(PD). A total of 23,417 and 389 peptides,as well as 15,146 and 75 collagen peptides were identified from AAE, AED and PD, respectively. Angiotensin converting enzyme (ACE) inhibitory activity,dipeptidyl peptidase IV(DPP-IV) inhibitory activity,prolyl endopeptidase(PEP) inhibitory activity and antioxidant activity were used to evaluate the bioactivities of AAE,AED and PD,and it was found that the sequence of their bioactivities was AAE<AED<PD. All the results above proved that AED released more collagen peptides and PD possessed better biological activities. It suggests that the two edible ways of antler are complemented each other and have their own advantages.
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Objective:To explore the effect of differentiation of cardiac stem cells(CSC)mediated by pilose antler polypeptides on the expressions of terminal myocardial differentiation genes atrial natriuretic polypeptide (ANP)and myosin light chain 2v(MLC-2v),and to clarify the mechanism of repairing the damaged myocardium.Methods:The healthy male Wistar rats born 2 d were selected to extract the CSC.The surface marker c-Kit and the purity of CSC were identified by immunofluorescence and flow cytometry.The culture dish was used as the experimental unit and the extracted cells were divided into the following four groups:blank control group(the same amount of buffer was added for induction),5-azacytidine group(induced with 3 μmol·L-15-azacytidine),pilose antler polypeptides group(induced with 800 mg·L-1pilose antler polypeptides)and combined group(induced with 800 mg·L-1pilose antler polypeptides and 3 μmol·L-15-azacytidine);the cells were incubated for 48 h in the condition of 37℃ and 5% CO2.The expression levels of ANP and MLC-2v in supernatant of the cells in various groups were detected by ELISA method.The expression levels of ANP and MLC-2v mRNA in the cells in various groups were detected by RT-PCR method.Results:CSC were prepared with the purity>95%.The results of ELISA showed that the expression levels of ANP and MLC-2v in 5-azacytidine,pilose antler polypeptides and combined groups were significantly increased compared with blank control group(P<0.05).The expression levels of ANP and MLC-2v in combined group were increased compared 5-azacytidine and pilose antler polypeptides groups,but there were no significant differences(P>0.05).The result of RT-PCR showed that the expression levels of ANP and MLC-2v mRNA in 5-azacytidine,pilose antler polypeptides and combined groups were significantly increased compared with blank control group(P<0.05).Conclusion:Pilose antler polypeptides can induce the differentiation of CSC into cardiac cells by promoting the expressions of ANP and MLC-2v,and they can reduce the cytotoxicity induced by 5-azacytidine in some degrees.
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Dermophytosis is a common fungal disease that affects fast-growing antlers of sika deer (Cervus nippon)and red deer (Cervus elaphus),causing the so-called 'white-skin antlers'and 'crusted antlers'.Here we described the features of dermophytosis in deer antler observed from 20 affected deer from 8 farms in Jilin and Liaoning province by clinical findings,he-matology,pathological examination and fungal species distribution.The fungal infection in the antlers as indicated by HE stai-ning,affected only epidermis and the dermis layers,with the main lesion of necrosis of the dermis tissue and inflammatory in-filtrate.Hematologic profile suggested the insignificant cell count change of lymphocyte,neutrophil,white blood cell between dermophytosis and healthy deer(n=10).A total of 68 fungi isolates were then recovered from the antlers with dermophytosis, of which 64.7% (44/68)were identified as members within Deuteromycotina,the rest 35.3% (24/68)belonged to the Saccha-romycotina.Notably,the well-known opportunistic pathogen,including species within Trichophyton,Epidermophyton as well as Candida albican,might account for the dermophytosis of deer antler.In conclusion,'white-skin antlers'and 'crusted antlers'are high likely caused by opportunistic fungi.
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To investigate the effects of anti-androgen drugs and melengestrol acetate (MGA) on development of regrowth antlers in 6 year old sika deer, twenty healthysika deerwith similar body weight and antler weightwere randomly divided into five groups by using single factor test design: flutamide (=4), bicalutamide (=4), progesterone acetate (CPA, =4), melengestrol acetate (MGA, =4), control(=4). All deer were fed with same diets and were housed outside together in an opened fence of 15 m×30 m with free access to water and feed. Treatment groups were injected subcutaneously sustained-release agents of the four drugs respectively when two-branched antlers were harvested. The control group had no special treatment. In the experiment period of 60 d, blood sampleswere collected for 4 times for each deer. The concentration of testosterone in plasma was tested and analyzed to compare the changes between different groups. Development of regrowth antlers was observed. At the end of the experiment, regrowth antlers were weighted and analyzed. The resultsshowed that the weights of regrowth antlers in treatment groups were significantly greater than those from control group and the weight gain (as compared with the control group) was 100.50%, 64.46%, 87.16% and 117.46% respectively in flutamide group, bicalutamide group, progesterone acetate group and melengestrol acetate group. For plasma testosterone concentration, it was not significantly different in the early stage (in the first 35 d), but at the end of the experimen, it was significantly higher than that of earlier stage (<0.01) in various groups. Testosterone concentration of flutamide treated group was significantly lower than that of the other groups (<0.01), while the level inbicalutamide and MGA treated groups was significantly higher than that in other groups (<0.01). The results showed that both anti-androgen drugs and MGA treatment promoted the development of regrowth antlers and increased the weight of regrowth antlers, where the effect was most significant by MGA treatment. From the morphological observation of the antlers, it was found that anti-androgen and MGA treatments prolonged the growth period of regrowth antlers through delaying the ossification of antlers. However, plasma testosterone concentration was not affected by the treatments.
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To investigate the chemical compositions of "antler powder" and "antler slice", two types of processed products of Cervi Cornu Pantotrichum (CCP) documented in Chinese Pharmacopoeia. With polysaccharides, crude protein, amino acids, fatty acids, mineral elements, biogenic amines, nucleosides and nucleobases as the evaluating indicators, the antler powder and antler slice processed with methods documented in Chinese Pharmacopoeia were compared in this study. The results showed that as compared with the antler powder by directly "chopping into pieces, and grinding into fine powder", the crude protein, amino acids, biogenic amines, nucleosides and nucleobases contents were reduced by 5.01%, 4.35%, 5.90%, 27.62% respectively in antler slices processed with 40% ethanol; the polysaccharides and nucleosides contents were reduced by 24.53% and 21.07% respectively in antler slices processed with 50% ethanol; and the crude protein and nucleosides contents were reduced by 1.65% and 20.52% in antler slices processed with 60% ethanol. While the contents of fatty acids and mineral elements were not decreased in these three methods. Polysaccharide, crude protein, amino acids, and nucleosides contents in "antler slices" were less than those in "antler powder", most notably in polysaccharides and nucleosides. According to the comprehensive scores of principal component analysis (PCA), the decrease of active ingredient determined in this study was lowest in antler slice processed with 50% ethanol.
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Objective: To explore the effect of differentiation of cardiac stem cells (CSC) mediated by pilose antler polypeptides on the expressions of terminal myocardial differentiation genes atrial natriuretic polypeptide (ANP) and myosin light chain 2v (MLC-2v), and to clarify the mechanism of repairing the damaged myocardium Methods: The healthy male Wistar rats born 2 d were selected to extract the CSC. The surface marker c-Kit and the purity of CSC were identified by immunofluorescence and flow cytometry. The culture dish was used as the experimental unit and the extracted cells were divided into the following four groups: blank control group (the same amount of buffer was added for induction), 5-azacytidine group (induced with 3 jumol · L-1 5-azacytidine), pilose antler polypeptides group (induced with 800 mg · L-1 pilose antler polypeptides) and combined group (induced with 800 mg · L-1 pilose antler polypeptides and 3 μmol · L-1 5-azacytidine); the cells were incubated for 48 h in the condition of 37°C and 5% CO2. The expression levels of ANP and MLC-2v in supernatant of the cells in various groups were detected by ELISA method. The expression levels of ANP and MLC-2v mRNA in the cells in various groups were detected by RT-PCR method. Results: CSC were prepared with the purity0.05). The result of RT-PCR showed that the expression levels of ANP and MLC-2v mRNA in 5-azacytidine, pilose antler polypeptides and combined groups were significantly increased compared with blank control group (P<0.05). Conclusion: Pilose antler polypeptides can induce the differentiation of CSC into cardiac cells by promoting the expressions of ANP and MLC-2v, and they can reduce the cytotoxicity induced by 5-azacytidine in some degrees.
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Objective: To discuss the protective effect of velvet antler polypeptides (VAP) combined with Schwann cells (SCs) modified by glial cell line derived neurotrophic factor (GDNF) gene on the apoptosis of spinal cord neurons induced by beta-amyloid 25-35 (Aβ25-35). Methods: The spinal cord cells of fetal mice were prepared and the spinal cord neurons in logarithmic growth phase were taken; the apoptosis of spinal cord neurons was induced by Aβ25-35. The spinal cord neurons were divided into normal cell group (the normal spinal cord neurons), induced apoptosis group (the apoptotic spinal cord neurons induced by Aβ25-35, SCs group (the apoptotic spinal cord neurons induced by Aβ25-35 + SCs), GDNF group (the apoptotic spinal cord neurons induced by Aβ25-35 + GDNF), SCs + GDNF group (the apoptotic spinal cord neurons induced by Aβ25-35 + GDNF-transfected SCs) and VAP combination group (the apoptotic spinal cord neurons induced by Aβ25-35 + VAP combined with GDNF-transfected SCs). Flow cytometry was used to detect the apoptotic rates of spinal cord neurons in various groups; immunohistochemical staining was used to detect the number of caspase-3 positive cells in the spinal cord neurons in various groups. Results; After suspension inoculation of fetal spinal cord neurons, most of them were round at the initial stage. The flow cytometry results showed that compared with induced apoptosis group, the apoptotic rates of spinal cord neurons in SCs, GDNF, SCs + GDNF, and VAP combination groups were decreased (P<0.05). Compared with SCs + GDNF group, the apoptotic rate of spinal cord neurons in VAP combination group was decreased (P<0. 05). The immunohistochemistry results showed that the expression of caspase-3 in spinal cord neurons in various groups could be found. There were no significant differences in the number of caspase-3 positive cells and cell staining between SCs group, GDNF group and SCs + GDNF group; but the number of caspase-3 positive cells in SCs group, GDNF group and SCs + GDNF group were significantly higher than that in induced apoptosis group (P<0. 05). Conclusion: VAP combined with GDNF-transfected SCs has the protective effect on the apoptosis of spinal cord cells by reducing the expression of caspase-3 in spinal cord neurons.