[Identification of Cervi Cornu (Cervus elaphus) and its formula granules based on PCR-RFLP].

Cervi Cornu is the ossified antler, or the base antler that falls off in the spring of the following year after the pilose antler is sawn off from Cervus elaphus or C. nippon, as a precious traditional Chinese medicine, has been recognized for its medicinal value and widely used in clinical practice. However, the origins of Cervi Cornu are miscellaneous, and Cervi Cornu is even mixed with adulterants in the market. Currently, there is a shortage of ways to identify Cervi Cornu and no standard to control the quality of Cervi Cornu. So it is valuable to develop a way to effectively identify Cervi Cornu from the adulterants. In this study, the differences in the mitochondrial barcode cytochrome b(Cytb) gene sequences of C. elaphus, C. nippon and their related species were compared and the specific single nucleotide polymorphism(SNP) sites on the Cytb sequences of Cervi Cornu were screened out. According to the screened SNPs, Cervi Cornu-specific primers dishmy-F and dishmy-R were designed. The PCR system was established and optimized, and the tolerance and feasibility of Taq polymerases and PCR systems affecting the repeatability of the PCR method were investigated. The amplification products of C. elaphus and C. nippon were digested using the restriction enzyme MseⅠ. The results showed that after electrophoresis of the product from PCR with the annealing temperature of 56 ℃ and 35 cycles, a single specific band at about 100 bp was observed for C. elaphus samples, and the product of C. elaphus samples was 60 bp shorter than that of C. nippon samples. There was no band for adulterants from other similar species such as Alces alces, Rangifer tarandus, Odocoileus virginianus, O. hemionus, Cap-reolus pygargus, Przewalskium albirostis and negative controls. The polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) method established in this study can quickly and accurately identify Cervi Cornu originated from C. elaphus in crude drugs, standard decoctions, and formula granules, and distinguish the origins of Cervi Cornu products, i.e., C. nippon and similar species. This study can be a reference for other studies on the quality standard of other formula granules of traditional Chinese medicines.

Minimizing pain in deer antler removal: Local anaesthetics in ZnO nanoparticle based collagen dressings as a promising solution.

Antler removal in deer is a common practice for various purposes, including meat production and traditional medicine. However, the current industry practice using lidocaine as a local anesthetic has limitations, such as short duration of action and the potential for postoperative infections. In this study, we investigated the performance of a ZnO collagen nanocomposites loaded with local anesthetics to improve wound management and alleviate pain associated with antler removal in red deer. The research involved the preparation of collagen nanocomposites with local anesthetics and testing the drug release rates using in vitro drug release tests. Pharmacokinetic analysis was performed to evaluate the total drug release from the collagen matrix in red deer after velvet removal. Additionally, the analgesic efficacy of these collagen nanocomposite dressings was assessed after antler removal in red deer. Functionalized ZnO nanoparticles were incorporated into collagen fibers to enhance their mechanical stability and prolong drug release. The developed collagen nanocomposites aimed to slowly release local anesthetics and promote wound healing. The findings of this research could have significant implications for improving the pain management and wound healing associated with antler removal in deer. The results obtained from the in vitro drug release tests, pharmacokinetic analysis, and analgesic efficacy evaluations provide valuable insights into the understanding and development of novel approaches for antler removal procedures in red deer. The findings contribute to the advancement of knowledge in this field and lay the foundation for future implementation of improved techniques and protocols for antler removal.

Structural characterisation of deer sinew peptides as calcium carriers, their promotion of MC3T3-E1 cell proliferation and their effect on bone deposition in mice.

Deer sinew as a by-product has high collagen and nutritional value. This study focuses on its hydrolysate being used as a calcium carrier to develop functional foods. The chelation mechanism was analyzed by SEM, EDS, UV-vis, FTIR, and fluorescence spectroscopy and zeta potential analysis after using peptide-sequenced deer sinew peptides for chelation with calcium ions. The results showed that the chelation of deer sinew peptides with calcium ions occurs mainly at the O and N atoms of carboxyl, amino and amide bonds. In vitro and in vivo studies revealed that deer sinew peptide-calcium chelate (DSPs-Ca) promoted the proliferation of MC3T3-E1 cells without toxic side effects and increased the alkaline phosphatase activity. The DSPs-Ca group improved the bone microstructure induced by low calcium, as well as up-regulated the expression of genes responsible for calcium uptake in the kidneys, as evidenced by serum markers, bone sections, bone parameters, and gene expression analyses in low-calcium-fed mice. From the above, it can be concluded that DSPs-Ca is expected to be a calcium supplement food for promoting bone health.

Muscone inhibits the progression of atherosclerotic plaques in mice aorta by inhibiting the NF-κB/p65 pathway.

Atherosclerosis (AS) is considered to be one of the main pathogenic factors of coronary heart disease, cerebral infarction and peripheral vascular disease. Oxidative stress and inflammation run through the occurrence and development of atherosclerosis and related cardiovascular events. Muscone is a natural extract of deer musk and also the main physiological active substance of musk. This study investigated the impact of muscone on atherosclerosis. ApoE-/- mice were used to establised AS model and injected with low-dose (4 mg/kg/day) or high-dose (8 mg/kg/day) of muscone intraperitoneally for 4 weeks. Then aortic tissues were collected, and pathological sections of the aorta were prepared for oil red staining, HE and masson staining. The changes of MDA, SOD, VCAM-1, NF-κB, and TNF-α were observed by Western blotting or immunofluorescence staining. The results showed that high-dose muscone could effectively reduce the plaque area/aortic root area and relative atherosclerotic area, reduce the collagen composition in plaque tissue. In addition, we also found that high-dose muscone can effectively increase MDA level, reduce the level of SOD, and inhibit the expression of VCAM-1, NF-κB/p65, TNF-α in arterial plaques. Our results indicate that the administration of muscone has the benefit of inhibiting atherosclerosis. The potential mechanisms may be associated with antioxidant effect and inhibition of inflammatory reaction in arterial plaques. With the increasing understanding of the relationship between muscone and atherosclerosis, muscone has high potential value as a new drug to treat atherosclerosis.

DNA barcoding for the identification and authentication of medicinal deer (Cervus sp.) products in China.

Deer products from sika deer (Cervus nippon) and red deer (C. elaphus) are considered genuine and used for Traditional Chinese Medicine (TCM) materials in China. Deer has a very high economic and ornamental value, resulting in the formation of a characteristic deer industry in the prescription preparation of traditional Chinese medicine, health food, cosmetics, and other areas of development and utilization. Due to the high demand for deer products, the products are expensive and have limited production, but the legal use of deer is limited to only two species of sika deer and red deer; other wild deer are prohibited from hunting, so there are numerous cases of mixing and adulteration of counterfeit products and so on. There have been many reports that other animal (pig, cow, sheep, etc.) tissues or organs are often used for adulteration and confusion, resulting in poor efficacy of deer traditional medicine and trade fraud in deer products. To authenticate the deer products in a rapid and effective manner, the analysis used 22 deer products (antler, meat, bone, fetus, penis, tail, skin, and wool) that were in the form of blind samples. Total DNA extraction using a modified protocol successfully yielded DNA from the blind samples that was useful for PCR. Three candidate DNA barcoding loci, cox1, Cyt b, and rrn12, were evaluated for their discrimination strength through BLAST and phylogenetic clustering analyses. For the BLAST analysis, the 22 blind samples obtained 100% match identity across the three gene loci tested. It was revealed that 12 blind samples were correctly labeled for their species of origin, while three blind samples that were thought to originate from red deer were identified as C. nippon, and seven blind samples that were thought to originate from sika deer were identified as C. elaphus, Dama dama, and Rangifer tarandus. DNA barcoding analysis showed that all three gene loci were able to distinguish the two Cervus species and to identify the presence of adulterant species. The DNA barcoding technique was able to provide a useful and sensitive approach in identifying the species of origin in deer products.

Surveillance of ticks on a tropical island with introduced ungulates.

The abundance of hard ticks can be determined by abiotic factors and the presence of suitable hosts. Since deer represent important hosts for many tick species, it is imperative to investigate whether deer introduction will increase the number of ticks, which can transmit a wide variety of pathogens. The sika deer (Cervus nippon) was introduced to Green Island, Taiwan, to supply velvet antlers, a traditional Chinese medicine. However, they were later released into the wild after a steep decline in antler prices. We conducted surveys for questing ticks, ticks on rodents and shrews, and the fecal pellet groups of ungulates (deer and goats) in 31 transects within 3 habitat types (forest, grassland, and roadside) on Green Island every 2 months in 2019. A total of 5,321 questing ticks were collected. All collected ticks were Haemaphysalis mageshimaensis Saito and Hoogstraal, a species first identified on an island with an abundance of sika deer. Additionally, 48 ticks collected from 126 trapped rodents and shrews were almost invariably Ixodes granulatus (except for 1 larval Haemaphysalis sp.). We did not find a positive correlation between the number of questing ticks and the number of fecal pellet groups, suggesting that factors other than ungulate abundance affect tick abundance. Nevertheless, large populations of questing ticks along some roadsides suggest a high risk of tick-borne diseases to tourists on this tropical island.

Gut Microbiota Combined with Metabolomics Reveal the Mechanisms of Sika Deer Antler Protein on Cisplatin-Induced Hepatorenal Injury in Mice.

Cisplatin is a widely used antineoplastic drug, though its adverse effects, particularly its hepatorenal toxicity, limit its long-term application. Sika deer antler is a valuable traditional Chinese medicine (TCM) documented to possess the capacity for tonifying the kidney and regulating the liver, of which the sika deer antler protein is an important active ingredient. In this study, two protein fractions, SVPr1 and SVPr2, of sika deer antler were purified and administered to mice treated with cisplatin, and serum metabolome and fecal microbiota were measured using ultrahigh-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) and 16S rRNA gene sequencing. SVPr1 and SVPr2 significantly ameliorated cisplatin-induced liver and kidney injury and reduced mitochondrial dysfunction, oxidative stress, inflammatory response, and apoptosis. In addition, SVPr1 and SVPr2 impacted the gut microbiota structure of mice, significantly increasing the relative abundances of Lactobacillus, which deserves to be scrutinized. Moreover, SVPr1 and SVPr2 antagonism of cisplatin-induced hepatorenal injury may be related to the regulation of lysine degradation, tryptophan metabolism, and riboflavin metabolism pathways, significantly altering the levels of L-saccharopine, L-lysine, L-kynurenine, 3-methylindole, xanthurenic acid, riboflavin, and D-ribulose-5-phosphate. A correlation between the differential metabolites and Lactobacillus was identified. These findings increased the knowledge of the gut microbiota-metabolites axis mediated by SVPr1 and SVPr2, and may be able to contribute to the development of new therapeutic strategies for the simultaneous prevention and treatment of liver and kidney injury from cisplatin treatment.

[The distribution of musk and river deer in local chronicles in Guizhou].

Some information and records about musk (She, ) and river deer (Zhang, /) can be found in local chronicles or documents in Guizhou. They were taken as the same species in terms of a medicinal animal. The records for their living areas in Guizhou were neither clear nor in detail in the Ming Dynasty, but were specific in the Qing Dynasty and more concise in the period of the Republic of China. The living areas for musk and river deer reduced from the Ming Dynasty to the Republic of China. Such change was believed to be the result of the natural environment and the social factors as well, such as the reclamation of mountain areas in Guizhou, the reduction of forests, and the demand and resulting exorbitant prices that led to excessive hunting.

Development of a tetra-primer ARMS-PCR for identification of sika and red deer and their hybrids.

Accurate identification of deer-derived components is significant in food and drug authenticity. Over the years, several methods have been developed to authenticate these products; however, identifying whether female deer products are hybrids is challenging. In this study, the zinc finger protein X-linked (ZFX) gene sequences of sika deer (Cervus nippon), red deer (Cervus elaphus) and their hybrid offspring were amplified and sequenced, the X221 and X428 species-specific single nucleotide polymorphisms (SNP) loci were verified, and a tetra-primer amplification refractory mutation system (T-ARMS-PCR) assay was developed to identify the parent-of-origin of female sika deer, red deer, and their hybrid deer. The T-ARMS-PCR developed based on the X221 locus could identify sika deer, red deer, and their hybrid offspring according to the presence or absence of PCR product sizes of 486 bp, 352 bp, and 179 bp, respectively, just as X428 locus could identify sika deer, red deer, and their hybrid offspring according to the presence or absence of PCR product sizes of 549 bp, 213 bp, and 383 bp, respectively. Forty products labeled deer-derived ingredients randomly purchased were tested using this assay, and the results showed that the identification results based on the two SNP loci were utterly consistent with the actual sources. In addition, this method was found to be accurate, simple, convenient, and with high specificity, thus providing an essential technical reference for deer product species identification. It is also an important supplement to the identification methods of the original ingredients of existing deer products.

Simplified detection of the species of origin of antler velvets using single-stranded tag hybridization chromatographic printed-array strip.

In this study, we developed a convenient and easy-to-use origin identification method for antler velvets based on a simple DNA extraction technique and single-stranded tag hybridization chromatographic printed-array strip (STH-PAS). The primer sets used to detect Cervus elaphus, Rangifer tarandus, and 12S rRNA did not engage in non-specific reactions such as primer dimer formation. In both the triplex and singleplex assays, the sensitivity was < 1 ng DNA. Moreover, Cervus elaphus DNA could be detected in OTC crude drug products. Although the detection sensitivity resulting from the simplified extraction was slightly lower than that obtained with extraction by conventional methods, the amount of DNA was sufficient even from a small sample. The choice of a triplex or singleplex assay will depend on the purpose of the test. For example, if it is important to determine whether the antler velvet is derived from Cervus elaphus or Rangifer tarandus, a triplex assay is appropriate. If it is necessary to explore whether antler velvet from Cervus elaphus is included in an OTC crude drug product, a singleplex assay using the Cervus elaphus primer set is informative. If it is necessary to explore whether powdered antler velvet includes counterfeit products (from Rangifer tarandus), a singleplex assay employing the Rangifer tarandus primer is appropriate. The singleplex assay detects minor components even at a 1,000:1 ratio. Our study thus demonstrated the utility of a method combining simple DNA extraction with STH-PAS for efficient identification of the origin of antler velvets.

[The content of zinc and selenium in local food products of Yakutia].

Deficiency of the necessary complex of micronutrients under abnormal nutrition structure creates a problem for the preservation of health and performance of the population. In this regard, it is very relevant to develop a science-based strategy for the consumption of traditional Yakut national food that have a high nutritional value and are able to satisfy the requirements of the human body in micronutrients. The purpose of the research was to investigate the content of selenium and zinc in the local foods which are most commonly consumed by the population of Yakutia. Material and methods. The objects of study were meat (7-9 cuts each) and offal (9-11 species each) of the Yakut cattle breed (2 bulls at the age of 2.5 years), of the Yakut horse foals (3 heads at the age of 6 months), northern domestic deer (3 heads), and whitefish (Coregonus muksun), Yakut crucian carp (Carassius carassius jacuticus), lake minnow [Phoxinus percnurus (Pallas)] (3 kg each). The trace elements, zinc and selenium, were determined by infrared spectroscopy. Results. The greatest amount of zinc in the meat of farm animals was noted in the meat of Yakut cattle (6.8±0.3 mg/100 g) and Yakut horse foals (6.7±0.2 mg/100 g), and the lowest - in the meat of domestic reindeer (1.5±0.1 mg/100 g). In terms of selenium content, the meat of domestic reindeer had the highest indicators (37.0±1.0 µg/100 g), and the meat of Yakut cattle had the lowest (19.0±0.8 µg/100g). The highest concentration of zinc and selenium was found in by-products of reindeer: zinc level in the heart and liver was 12.8 mg/100 g, in the small intestine and rennet - 19.0-20.4 mg/100 g, selenium level in the colon and rennet was 41.0-46.7 µg/100 g. The content of zinc and selenium in the fresh-water muksun belly (2.14±0.08 mg and 45.0±1.8 µg in 100 g) was 32.3-37.2% higher than in the fillet of muksun, and selenium level was 3 fold higher than in the Yakut carp and in the lake minnow. The consumption of 100-200 g of meat or by-products of Yakut cattle, Yakut horse foals, by-products of reindeer or Yakut crucian carp can fully cover the daily requirement of an adult in zinc. The consumption of 200 g of venison or muksun completely covers the daily need for selenium, while the portion of the other studied products contains about half or more of recommended daily intake of this trace element. Conclusion. The data presented in the article show that the population of Yakutia, with a rational diet with local products, can meet the requirements in selenium and zinc in accordance with physiological needs.

Ancient human DNA recovered from a Palaeolithic pendant.

Artefacts made from stones, bones and teeth are fundamental to our understanding of human subsistence strategies, behaviour and culture in the Pleistocene. Although these resources are plentiful, it is impossible to associate artefacts to specific human individuals1 who can be morphologically or genetically characterized, unless they are found within burials, which are rare in this time period. Thus, our ability to discern the societal roles of Pleistocene individuals based on their biological sex or genetic ancestry is limited2-5. Here we report the development of a non-destructive method for the gradual release of DNA trapped in ancient bone and tooth artefacts. Application of the method to an Upper Palaeolithic deer tooth pendant from Denisova Cave, Russia, resulted in the recovery of ancient human and deer mitochondrial genomes, which allowed us to estimate the age of the pendant at approximately 19,000-25,000 years. Nuclear DNA analysis identifies the presumed maker or wearer of the pendant as a female individual with strong genetic affinities to a group of Ancient North Eurasian individuals who lived around the same time but were previously found only further east in Siberia. Our work redefines how cultural and genetic records can be linked in prehistoric archaeology.

Correlation between airborne pollen data and the risk of tick-borne encephalitis in northern Italy.

Tick-borne encephalitis (TBE) is caused by a flavivirus that infects animals including humans. In Europe, the TBE virus circulates enzootically in natural foci among ticks and rodent hosts. The abundance of ticks depends on the abundance of rodent hosts, which in turn depends on the availability of food resources, such as tree seeds. Trees can exhibit large inter-annual fluctuations in seed production (masting), which influences the abundance of rodents the following year, and the abundance of nymphal ticks two years later. Thus, the biology of this system predicts a 2-year time lag between masting and the incidence of tick-borne diseases such as TBE. As airborne pollen abundance is related to masting, we investigated whether inter-annual variation in pollen load could be directly correlated with inter-annual variation in the incidence of TBE in human populations with a 2-year time lag. We focused our study on the province of Trento (northern Italy), where 206 TBE cases were notified between 1992 and 2020. We tested the relationship between TBE incidence and pollen load collected from 1989 to 2020 for 7 different tree species common in our study area. Through univariate analysis we found that the pollen quantities recorded two years prior for two tree species, hop-hornbeam (Ostrya carpinifolia) and downy oak (Quercus pubescens), were positively correlated with TBE emergence (R2 = 0.2) while a multivariate model with both tree species better explained the variation in annual TBE incidence (R2 = 0.34). To the best of our knowledge, this is the first attempt at quantifying the correlation between pollen quantities and the incidence of TBE in human populations. As pollen loads are collected by widespread aerobiological networks using standardized procedures, our study could be easily replicated to test their potential as early warning system for TBE and other tick-borne diseases.

Decreased Immunoreactivity of Hepatitis E Virus Antigen Following Treatment with Sakhalin Spruce (Picea glehnii) Essential Oil.

The hepatitis E virus (HEV) causes a common infectious disease that infects pigs, wild boars, deer, and humans. In most cases, humans are infected by eating raw meat. Some essential oils have been reported to exhibit antiviral activities. In this study, in order to investigate the anti-HEV properties of essential oils, the immunoreactivities of HEV antigen proteins against the relevant antibodies were analyzed after the HEV antigens underwent treatment with various essential oils. The essential oils extracted from the tea tree, which was previously reported to exhibit antiviral activity, lavender, and lemon had strongly reduced activity. We found that treatment with the essential oil prepared from Sakhalin spruce was associated with the strongest reduction in immunoreactivity of HEV antigen protein(s) among the tested substances. The main volatile constituents of Sakhalin spruce essential oil were found to be bornyl acetate (32.30 %), α-pinene (16.66 %), camphene (11.14 %), camphor (5.52 %), ß-phellandrene (9.09 %), borneol (4.77 %), and limonene (4.57 %). The anti-HEV properties of the various components of the essential oils were examined: treatment with bornyl acetate, the main component of Sakhalin spruce oil, α-pinene, the main component of tea tree oil, and limonene, the main component of lemon oil, resulted in a strong reduction in HEV antigen immunoreactivity. These results indicate that each main component of the essential oils plays an important role in the reduction of the immunoreactivity of HEV antigen protein(s); they also suggest that Sakhalin spruce essential oil exhibits anti-HEV activity. In a formulation with the potential to eliminate the infectivity of HEV in foodborne infections, this essential oil can be applied as an inactivating agent for meat processing and cooking utensils, such as knives and chopping boards.

Pharmacodynamic structure of deer antler base protein and its mammary gland hyperplasia inhibition mechanism by mediating Raf-1/MEK/ERK signaling pathway activation.

Mammary gland hyperplasia (MGH) is a common mammary disease whose main pathogenesis is the disruption of estradiol (E2) and progesterone (P) secretion, thereby causing overproliferation of mammary epithelial cells and mammary gland tissue hyperplasia. Deer antler base is a traditional Chinese medicine that has been used for many years to treat MGH. However, its pharmacological mechanism and pharmacodynamic material basis are unclear. In this study, we for the first time used the graded salting method to classify deer antler base protein (CNCP) as CNCP-A, CNCP-B, and CNCP-C and explored the pharmacological mechanism of the anti-MGH properties of CNCP. We found that CNCP could regulate the hormonal levels of E2, P, and follicle stimulating hormone (FSH) and improve the histopathological condition. The potential mechanism might be related to the recombinant C-Raf proto oncogene serine/threonine protein kinase/mitogen-activated protein/extracellular regulated protein kinase (Raf-1/MEK/ERK) signaling pathway. By upregulating the protein expression of the follicle stimulating hormone receptor (FSHR), cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) inhibited the activation of the downstream Raf-1/MEK/ERK signaling pathway, which in turn inhibited the proliferation of mammary epithelial cells. We analyzed the physicochemical properties of CNCP-A, CNCP-B, and CNCP-C and obtained CNCP-C-I by column chromatographic purification of the best pharmacophore protein CNCP. Using high-performance liquid gel filtration chromatography (HPGFC), we determined the molecular weight of CNCP-C-I and identified it by high-performance liquid tandem mass spectrometry (LC-MS/MS) to obtain the first match for a high confidence protein KRT1. This study provides a theoretical basis for the development of effective traditional Chinese medicines with low toxicity levels for the prevention and treatment of mammary gland diseases.

Discovery of Elaphuri Davidiani Cornu-specific peptide biomarkers by peptidomics analysis-based method.

Elaphuri Davidiani Cornu (EDC) is the antler of the male Père David's deer, which has been reported to have multiple biological activities, and its use as a traditional Chinese medicine (TCM) in China has been known for thousands of years. However, EDC is difficult to distinguish from other related species-derived antlers in powder or extract form in TCM clinic use, such as Cervus elaphus Cornu (CEC) and Cervus nippon Cornu (CNC), both derived from Cervidae and easily confused with EDC. In this study, a strategy using peptidomics combined with mathematics set analysis was used to identify EDC-specific peptide biomarkers, and four specific peptide biomarkers (Pep-E1-E4) were identified and validated. Pep-E1, Pep-E3, and Pep-E4 could be exclusively detected in EDC samples, with relative peak areas of 0.298 ± 0.060, 0.039 ± 0.015, and 0.037 ± 0.008, whereas Pep-E2 showed relative peak area of 0.516 ± 0.101 in EDC, 0.132 ± 0.026 in CEC, and 0.136 ± 0.047 in CNC samples, respectively. These four peptides are applicable to distinguish EDC from CEC and CNC, which is of great significance for the quality control of EDC.

Selenium and mercury concentrations, Se:Hg molar ratios and their effect on the antioxidant system in wild mammals.

The aim of this study was to a) evaluate the concentration of Se and Hg and their relationship in the tissues of 4 species of wild mammals, including Se:Hg molar ratios, and b) evaluate the effect of the analysed elements and their mutual proportions expressed as Se:Hg molar ratio, on the antioxidant system in the tissues of the tested animals. The study was performed on 31 animals belonging to four species: wild boar, red fox, roe deer, brown hare. Determination of Hg in liver, kidney and muscle of animals was performed using an AMA 254 mercury analyser. Total Se concentrations were determined using the spectrofluorometric method. In omnivores demonstrated higher Se concentrations in all analysed organs compared to the herbivores. The highest concentration of Hg was found in the kidneys of the tested animals, and the lowest in the muscles. High and moderate correlation between Se and Hg was observed in the liver of omnivorous, while in herbivores this correlation was weak. In all analysed samples, the Se:Hg molar ratios were above 1 (min: liver 5.9, max: kidney 110). Generally, the highest Se:Hg ratio values were found in kidney and the lowest in liver of tested animals. No significant correlation was found between GPx, GST and SOD activity and Se or Hg concentration in analysed organs. But it was observed that Se:Hg molar was negatively correlated with CAT activity in the most samples. The obtained results may suggest that omnivorous animals demonstrate greater Hg sequestration in the liver than herbivores, which has been proposed as one of the mechanisms of Se antagonistic action towards Hg. The ratio between Se and Hg, rather than the concentration of these elements in organs, affected the antioxidant status in the animal organism, specifically the CAT activity.

Effects of immunocastration and supplementary feeding level on the performance and blood biochemical markers of farmed yearling fallow deer (Dama dama).

In cervids, blood biochemical markers may reflect changes in various physiological and environmental factors, especially in response to changes in metabolism following nutrient supplementation or the manipulation of hormone production. Decreasing androgen production through immunocastration (IC) to ease the husbandry of male animals is currently a more ethically acceptable method than physical castration, but its commercial use is unexplored in fallow deer. Forty yearlings male fallow deer were grouped into four treatment combinations: IC on high (200 g commercial pellets + 600 g concentrate mixture of 90% oats and 10% wheat grains) or low (100 g commercial pellets + 300 g concentrate mixture of 90% oats and 10% wheat grains) level of feed supplementation, or noncastrated bucks on a high or low level of feed supplementation. Immunocastrated animals were vaccinated at the start of the study (Week 1) and again during Week 3 of the study. Diet affected all body growth parameters (slaughter weight, daily gain, carcass weight, dressing percentage and body condition score). Fallow deer from all treatments showed increasing concentrations of fat and energy blood biochemical markers over the study period, including plasma glucose (GLU) and triglyceride (TRIG), and decreased cholesterol (CHOL) and lipase (LIPA) concentrations. The higher level of supplementary feeding decreased plasma albumin (ALB) and creatinine (CREA), and increased globulin (GLOB) concentrations. On the other hand, IC and lower-level supplementation reduced growth performance. Overall, IC may be an interesting tool for welfare management of yearling stags for slaughter; however, the advantage appears to only be in well-fed animals, as low-level of feeding can further reduce growth performance in immunocastrated animals. Further studies should evaluate the carcass performance of animals under similar treatment conditions to ascertain the effects on muscle and fat yields.

Pilose antler (Cervus elaphus Linnaeus) polysaccharide and polypeptide extract inhibits bone resorption in high turnover type osteoporosis by stimulating the MAKP and MMP-9 signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Pilose antler is a traditional Chinese medicine used to improve kidney function, strengthen tendons and bones, and prolong life, among other uses. It is widely employed in the treatment of osteoporosis. However, the molecular mechanisms underlying the treatment of high turnover osteoporosis are not fully understood. AIM OF THE STUDY: The present study aimed to investigate the molecular mechanism underlying pilose antler polysaccharide and polypeptide extracts in inhibiting bone resorption in high turnover osteoporosis, and compare the effects of the two components alone and in combination to explore whether they could produce synergistic enhancement effects. MATERIALS AND METHODS: The quantitative and qualitative characteristics of pilose antler polysaccharide and polypeptide extracts were detected by UV-visible spectrophotometry and high-performance liquid chromatography. A rat model of retinoic acid-induced osteoporosis was used to evaluate the inhibitory effect of the extracts on bone resorption. Enzyme-linked immunosorbent assay (ELISA) was used to detect the activity of factors related to high turnover type osteoporosis in rat serum. Western blotting was used to detect the expression of proteins related to the MAKP and MMP-9 signaling pathways in rat femurs. Fluorescence quantitative PCR was used to detect the transcription levels of genes related to the MAKP and MMP-9 signaling pathways in rat femur tissues. Hematoxylin and eosin staining were used to observe the osteoprotective effects of pilose antler polysaccharides and polypeptides. RESULTS: The yield of pilose antler polysaccharides was 8.3%, and was mainly composed of mannose, glucosamine hydrochloride, glucuronic acid, Galacturonic acid, Galactose hydrochloride, glucose, and galactose. The yield of the polypeptides was 26.2%, and eighty percent of the molecular weight of the antler polypeptides was 1.6 kDa-7kD, among which, the molecular weight of 7kD peptide accounted for 52% of the total. Both polysaccharides and peptides could reduce the activities of TRACP, OCN, ERK1, JNK, and MMP-9 in rat serum and reduce both the protein expression and gene transcription levels of ERK1, JNK, and MMP-9 in rat femur tissue with significant differences compared with the model group. Both extracts exerted significant protective effects on rat femur tissue. The effect of pilose antler polypeptides alone was better than that of polysaccharides either alone or in combination. CONCLUSIONS: Pilose antler polysaccharides, polypeptides, and their mixtures could inhibit the occurrence of bone resorption of high turnover osteoporosis by stimulating the MAKP and MMP-9 signaling pathways to reduce the expression of the ERK1, JNK, and MMP-9 genes and proteins, and could help alleviate bone loss caused by retinoic acid. Pilose antler polypeptides had a stronger effect on inhibiting bone resorption. The combination of the two components did not show synergistic enhancement effect, and the polysaccharide tended to moderate the inhibitory enhancement effect of the polypeptide.