Protective effect of Sika Deer bone polypeptide extract on dexamethasone-induced osteoporosis in rats

BACKGROUND: Osteoporosis attacks approximately 10% of the population worldwide. Sika Deer (Cervus nippon), one of China's precious traditional medicinal animals, has been widely recorded in ancient Chinese medical books and claimed for centuries to have numerous medical benefits including bone strengthening. This study aimed to find the use of Sika Deer bone in treating osteoporosis according to traditional records and to investigate the protective effect of Sika Deer bone polypeptide extract on glucocorticoidinduced osteoporosis (GIOP) in rats. RESULTS: Sika Deer bone polypeptide extract could increase serum Ca2+ and BGP, decrease serum P3+, ALP, PTH, and CT, but had no effect on serum NO in rats with GIOP. The immunohistochemical iNOS results of the rats' distal femur were negative in each group. Besides the model group, the eNOS color reaction in osteoblasts was strongly positive in the other three groups. CONCLUSIONS: Sika Deer bone polypeptide extract can improve pathological changes in the microstructure and stimulate the expression of eNOS in osteoblasts. The protective effect on bone might be mediated by eNOS-dependent NO generation.

Identification of Cervi Cornu Pantotrichum and Cervi Cornu by Allele-specific PCR / 中国实验方剂学杂志

Objective: To obtain a rapid,efficiency and convenient polymerase Chain reaction(PCR) identification method for medicinal Cervi Cornu Pantotrichum,Cervi Cornu and its common adulterates. Method: Based on three single nucleotide polymorphisms (SNP) of Cytb gene DNA sequences among Cervus nippon,C. elaphus and its adulterants,a pair of species-specific primers (LR-238.F and LR-238.R) was designed,the reaction conditions were optimized,and the PCR method for identification was explored and verified in terms of tolerance and feasibility. Result: Through the established allele-specific PCR method,under the annealing temperature of 56℃ and cycle number of 35,250 bp of fragments were amplified from DNA templates of Cervi Cornu Pantotrichum,Cervi Cornu and its subspecies in origin animal samples as well as herbal medicines. All of the adulterants species of Przewalskium albirostris,Cervus eldi,Odocoileus hemionus,Dama dama,Alces alces,Elaphurus davidianus,Capreolus pygargus,Rusa unicolor and Rangifer tarandus were negative by the PCR assay. Conclusion: The identification primer is highly specific,and the allele-specific PCR identification method established in this paper can accurately identify the medicinal Cervi Cornu Pantotrichum and Cervi Cornu.

Identification of deer blood by PCR-RFLP method / 中草药

Objective To establish a polymerase chain reaction coupled with restriction fragment length polymorphism (PCR-RFLP) method which can identify deer blood. Methods We analyzed DNA sequences of cytochrome b (cytb) from different species (i.e. sika deer, red deer, pig, ox, sheep, and duck) and selected two DNA restriction enzymes of EcoRV and MseI to distinguish deer from other species and to sika deer from red deer, respectively. Genomic DNA of every sample was extracted by blood genomic DNA extraction kit. After PCR of cytb fragment, digestion by EcoRV or MseI was conducted and the results were analyzed. The capacity of this method to identify different proportional blood mix from different species also was determined. Results Using the PCR-RFLP method, EcoRV digestion made two fragments of 287 bp and 185 bp in deer, but no digestion in other species; MesI digestion made two fragments of 281 bp and 191 bp in sika deer, but three fragments of 281 bp, 126 bp, and 65 bp in red deer. The 191 bp fragment was a characteristic marker of sika deer, and the 126 bp was for red deer. The minimum detected proportion added to blood of deer with other sample was 3%, 6% of sika deer with red deer. Conclusion A PCR-RFLP method according to the sequences of cytb gene is established. This method can identify the blood of sika deer rapidly and conveniently.

Identification of Cervus nippon, C.elaphus and their hybridize samples based on COI and SRY gene / 中国中药杂志

For rapid identification of Cervus nippon, C. elaphus and their hybridize samples, the specific PCR for mutual authentication of them was established based on the SNPs in COI and SRY sequence. C. nippon, C. elaphus and their hybridize samples were collected from different origins, total DNA of 24 identified samples were extracted, and the COI and SRY gene was seqenced. SNPs in the COI and SRY sequences of the samples were found by Clustul X 2.1 program. Primers for identifying C. nippon and C. elaphus were designed according to the SNP site, two multi-PCR reaction system were established to identify them. In addition, 24 samples which were randomly collected in different herbal medicine market were identified. The band special for C. nippon (232 bp)and band special for C. elaphus (518 bp) based on COI sequence,and the band special for C. nippon (803 bp)and band special for C. elaphus (425 bp) based on SRY sequence, were found using multi-PCR reaction, and three of the twenty-four samples were identified as the hybridize samples. The multi-PCR reaction system could be used to identify C. nippon, C. elaphus and their hybridize samples.

Sudden death caused by diaphragmatic rupture following rib fracture in a female sika deer (Cervus nippon) at a zoo

A one-year-old female sika deer died suddenly with no preliminary signs during exhibition at a zoo. At necropsy, the carcass was emaciated and had dried fur. Examination of the thoracic cavity revealed a diaphragmatic rupture measuring 2 cm in diameter and a fracture in the middle of the right eighth rib. The liver and lungs had irregular circular discolorations caused by diaphragmatic rupture and subsequent herniation. Dark-brown-colored ascitic fluid, hydrothorax, and yellowish hydropericardium were also observed. The cause of death was determined to be diaphragmatic rupture caused by a rib fracture, which led to respiratory imbalance and circulatory disorders.

Electron microscopic observations on changes of cytoplasmic organelles in the oocytes of Cervus nippon hortulorun / 解剖学报

Objective Revealing the developmental regulation of Cervus nippon's oocyte and organelles. Methods In the experiment,follicle systems during both estrum and non- estrum were divided into the primordial follicle,growing follicle and mature follicle according to the Cervus nippon's follicle diameter size,formation of zonapellucida,appearance time of follicular cavity.At the same time,observations on cytoplasmic organelles in development of oocytes were conducted with electron microscopic,eyepiece micrometer and photomicrographic technique(The number of every oocyte observed is 6-8). Results In the primordial follicle and early growing follicle phase,the quantity of mitochondria,golgi apparatus,smooth endoplasmic reticulum and cortical granules increased gradually and all organelles moved to the cortical area.However,in the late growing follicle and mature follicle phase,Golgi apparatus and rough endoplasmic reticulum disappeared,cortical granules began to arrange themselves in line beneath the plasma membrane of the oocyte,mitochondrias dispersed toward the central region of cytoplasm,and almost all the round mitochondria with rare cristae turned into hooded ones, and nucleus compaction occurred.In addition,the short and thick microvilli began to appear from the primary follicle ovocyte,become intensive and slender when secondary follicle's ovocyte;It's until tertiary follicle's ovocyte,microvilli started to shorten and become coarse,and even parts of them contract from the zona pellucida gradually.Conclusion In the development of oocytes, the changes of type,quantity and distribution on mitochondria has a close relation with cells at proliferation,differentiation and metabolism level.Cortical granule has no association with golgi apparatus basically,but smooth endoplasmic reticulum(SER).The nuclei are the sites of RNA synthesis and warehouses,and its densification is the premise of meiosis recovery.