Calcium-chelating peptides from rabbit bone collagen:characterization,identification and mechanism elucidation

This study aimed to characterize and identify calcium-chelating peptides from rabbit bone collagen and explore the underlying chelating mechanism.Collagen peptides and calcium were extracted from rabbit bone by instant ejection steam explosion(ICSE)combined with enzymatic hydrolysis,followed by chelation reaction to prepare rabbit bone peptide-calcium chelate(RBCP-Ca).The chelating sites were further analyzed by liquid chromatography-tandem mass(LC-MS/MS)spectrometry while the chelating mechanism and binding modes were investigated.The structural characterization revealed that RBCP successfully chelated with calcium ions.Furthermore,LC-MS/MS analysis indicated that the binding sites included both acidic amino acids(Asp and Glu)and basic amino acids(Lys and Arg),Interestingly,three binding modes,namely Inter-Linking,Loop-Linking and Mono-Linking were for the first time found,while Inter-Linking mode accounted for the highest proportion(75.1%),suggesting that chelation of calcium ions frequently occurred between two peptides.Overall,this study provides a theoretical basis for the elucidation of chelation mechanism of calcium-chelating peptides.

Genome-edited rabbits:Unleashing the potential of a promising experimental animal model across diverse diseases

Animal models are extensively used in all aspects of biomedical research,with substantial contributions to our understanding of diseases,the development of pharmaceuticals,and the exploration of gene functions.The field of genome modification in rabbits has progressed slowly.However,recent advancements,particularly in CRISPR/Cas9-related technologies,have catalyzed the successful development of various genome-edited rabbit models to mimic diverse diseases,including cardiovascular disorders,immunodeficiencies,agingrelated ailments,neurological diseases,and ophthalmic pathologies.These models hold great promise in advancing biomedical research due to their closer physiological and biochemical resemblance to humans compared to mice.This review aims to summarize the novel gene-editing approaches currently available for rabbits and present the applications and prospects of such models in biomedicine,underscoring their impact and future potential in translational medicine.

Betaine addition to the diet alleviates intestinal injury in growing rabbits during the summer heat through the AAT/mTOR pathway

Background The aim of this experiment was to investigate the effect of different levels of betaine(Bet)inclusion in the diet on the intestinal health of growing rabbits under summer heat.A total of 100 weaned Qixing meat rabbits aged 35 d with body weight of 748.61±38.59 g were randomly divided into 5 treatment groups:control group(basal diet)and Bet groups(basal diet+0.75,1.0,1.5 or 2.0 g/kg Bet).The average daily temperature in the rabbitry during the experiment was 30.48°C and the relative humidity was 69.44%.Results Dietary addition of Bet had no significant effect on growth performance and health status of growing rabbits(P>0.05),but it increased ileal secretory immunoglobulin A content compared to the control under summer heat(P<0.05).Addition of 0.75 g/kg Bet up-regulated jejunal IL-4,down-regulated ileal TNF-αexpression(P<0.05).The addition of 1.0 g/kg Bet increased the villi height(VH)in the jejunum(P<0.05).Serum glucose levels were reduced,and the expression of SLC6A20 was up-regulated in jejunum and ileum of rabbits fed with 1.5 g/kg Bet(P<0.05).When added at 2.0 g/kg,Bet reduced serum HSP70 content,increased jejunal VH,and up-regulated duodenal SLC7A6,SLC38A2,mTOR and 4EBP-2 expression(P<0.05).Correlation analysis revealed that intestinal mTOR expression was significantly and positively correlated with SLC7A6,SLC38A2,SLC36A1 and IL-4 expression(P<0.05).Conclusions Dietary addition of Bet can up-regulate the expression of anti-inflammatory factors through the AAT/mTOR pathway,improve the intestinal immune function,alleviate intestinal damage in growing rabbits caused by summer heat,and improve intestinal health.

An Improved Artificial Rabbits Optimization Algorithm with Chaotic Local Search and Opposition-Based Learning for Engineering Problems and Its Applications in Breast Cancer Problem

Artificial rabbits optimization(ARO)is a recently proposed biology-based optimization algorithm inspired by the detour foraging and random hiding behavior of rabbits in nature.However,for solving optimization problems,the ARO algorithm shows slow convergence speed and can fall into local minima.To overcome these drawbacks,this paper proposes chaotic opposition-based learning ARO(COARO),an improved version of the ARO algorithm that incorporates opposition-based learning(OBL)and chaotic local search(CLS)techniques.By adding OBL to ARO,the convergence speed of the algorithm increases and it explores the search space better.Chaotic maps in CLS provide rapid convergence by scanning the search space efficiently,since their ergodicity and non-repetitive properties.The proposed COARO algorithm has been tested using thirty-three distinct benchmark functions.The outcomes have been compared with the most recent optimization algorithms.Additionally,the COARO algorithm’s problem-solving capabilities have been evaluated using six different engineering design problems and compared with various other algorithms.This study also introduces a binary variant of the continuous COARO algorithm,named BCOARO.The performance of BCOARO was evaluated on the breast cancer dataset.The effectiveness of BCOARO has been compared with different feature selection algorithms.The proposed BCOARO outperforms alternative algorithms,according to the findings obtained for real applications in terms of accuracy performance,and fitness value.Extensive experiments show that the COARO and BCOARO algorithms achieve promising results compared to other metaheuristic algorithms.

Effects of Incorporating Mango (Mangifera indica), Avocado (Persea americana) and Guava (Psidium guajava) Leaves Powder in the Ration on Growth Characteristics in Rabbits

A study conducted on burgundy rabbits evaluated the effects of rations containing mango (Mangifera indica), avocado (Persea americana) and guava (Psidium guajava) powder. For this purpose, 36 rabbits (20 males and 16 females) aged 60 to 70 days with an average weight of 1012 ± 133 g were divided randomly into 4 equal groups (T0, T1, T2 and T3) of 9 animals each according to body weight (bw). The rabbits were divided throughout the trial, the animals were fed, those of group T0 (control) with a standard feed without any type of leaf powder, while those of groups 1, 2 and 3 received the standard feed supplemented with Mangifera indica, Persea americana and Psidium guajava leaves powder at the rate of 0.5% respectively. After two months of treatment, the animals were fasted for 12 hours (20 h - 8 h) and sacrificed to evaluate carcass and digestive organ weights. The results revealed that the values of feed intake, live weight and weight gain were non-significantly (p > 0.05) increased in all animals regardless of the ration in reference to the control. The different leaves powder induced an increase in carcass yield in rabbits but this increase was only significant (p M. indica leaf powder (51.03 ± 0.24) as compared to the control (48.63 ± 0.74). Gut length was significantly increased in animals fed rations containing Psidium guajava and Persea americana leaves powder compared to the control. A significant increase in gut weight was observed in animals fed the Persea americana leaf supplemented ration. In general, mango leaf powder performed better than other types of powders.

Effects of Carrot (Daucus carota) Leaves Powder on Reproductive and Growth Characteristics of Rabbit (Oryctolagus cuniculus) Does

The present study was conducted to evaluate the effects of carrot leaf powder in the diet on reproductive and growth characteristics in rabbits. Thirty sexually matured rabbits does of 8 months old, 3.00 to 3.20 kg, were divided into three groups of 10 rabbits each, which were comparable in terms of body weight. Throughout the experiment, the rabbits of group 1 (control group) were fed ad libitum with a feed not containing carrot leaf powder (control ration), while those of groups 2 and 3 received the control ration supplemented with carrot leaf powder in concentrations of 0.5% and 1% respectively. Data was collected on feed intake, weight gain, receptivity, fertility, gestation length of does, litter size, weight, viability and sex ratio of the young rabbits till 5 weeks post-partum. There was no significant (P > 0.05) difference in the fertility rate, gestation duration and sex ratio of the kits with reference to the control. Litter size at birth was not significantly (P > 0.05) influenced in does fed diets containing carrot leaf powder compared to the control. However, the reversed effects were observed at five weeks post-partum. The live weight of weaned kits and weight gain of rabbit does at birth were significantly (P < 0.05) increased in treated rabbits compared to the control. The supplementation of carrot leaf powder in feed can improve production performance in female rabbits, through their antioxidant activities thanks to their various components, thus fighting against oxidative stress effects to which animals are exposed both from endogenous and environmental sources.

Methods to Reduce the Hypoglycemic Mortality of Alloxan in Diabetic Rabbit Model

Objective: To explore an intervention method to reduce the mortality of alloxan diabetes model, and to preliminarily analyze the mechanism of alloxan induced animal death. Methods: Healthy New Zealand rabbits were randomly divided into injection group, control group, experimental group and blank group. The single injection group was injected with 100 mg/kg alloxan once. The control group was given 5% glucose solution and 100 mg/kg alloxan was injected in two times. The experimental group was given 5% glucose solution orally, 100 mg/kg alloxan, 7 mL 0.9% NaCl intravenously and 5 mL 5% glucose intraperitoneally immediately, and blood glucose was continuously monitored, 10 mL 5% glucose intravenously and 10 mL 5% glucose intraperitoneally every 4 h in the hypoglycemic stage. The blank group does nothing. Liver and kidney tissues at different time periods were stained with HE and organ index was evaluated. Results: 1) A single injection of 100 mg/kg alloxan without any intervention resulted in 100% mortality. Before modeling, oral administration of 5% glucose solution, divided into two injections of 100 mg/kg alloxan, mortality reached 100%;A single injection of 100 mg/kg alloxan and continuous intervention of normal saline and glucose for 20 h can significantly reduce the mortality of alloxan induced diabetic rabbit model. 2) Liver and kidney tissues were damaged in different degrees at different time periods, and liver and kidney indexes were significantly increased after alloxan injection compared with the normal group, with statistical significance (P > 0.05). Conclusion: 1) Every 4 hours of hypoglycemia, 10 ml 5% glucose was injected intravenously 10 ml 5% glucose intraperitoneally. It can reduce the death rate of alloxan diabetic rabbit model and shorten the time of blood glucose measurement. 2) After the injection of alloxan, acute lesions of liver and kidney may occur in different degrees, or one of the causes of acute death of experimental animals.

Chemical Composition of Pineapple Press Residues (Ananas comosus) and Effect on Growth Performance of Rabbit (Oryctolagus cuniculus)

With a view to improving rabbit production performance, a trial on the chemical composition of pineapple press residue (Ananas comosus) and the effect of its incorporation in the ration on rabbit growth performance (Oryctolagus cuniculus) was carried out at the KUATE Cunicole Farm in Bandjoun, in Western Cameroon. To do this, 36 rabbits of the local breed, aged 53 days with an average weight of 1337 ± 119 g were distributed and randomly assigned to 3 experimental rations corresponding respectively to treatments or batches T0, T1 and T2. The animals in treatment T0 received a ration containing no pineapple press residue, while those in treatments T1 and T2 received a ration containing 20% and 40% pineapple press residue, respectively. These residues were dried and ground for chemical composition analysis. The feed served as well as refusals from the previous day were weighed each morning to assess feed intake. The animals were weighed every 7 days to assess weight performance. At the end of the trial which lasted 7 weeks, the animals were fasted for 24 hours, then sacrificed to evaluate carcass characteristics and the relative weights of some digestive organs. The results of this study showed that pineapple press residues had a high crude fiber content (19.2%) and energy (2500 Kcal/kg DM). Their incorporation had no significant effect on feed intake and feed conversion ratio. The average live weight, weight gain and average daily weight gain of the animals receiving the ration with 20% inclusion of pineapple press residue were comparable to those of the control group and significantly higher than those of animals fed with 40% inclusion of pineapple residue. The highest carcass yields were obtained with rabbits fed 20% pineapple press residue in their ration. The cost of feed for the production of a kilogram live weight of rabbit tends to decrease with the ration incorporated with 20% pineapple press residue. Pineapple press residues constitute a by-product that can be recycled and their incorporation at 20% can increase rabbit growth performance and reduce production costs.

Intravitreal slow-release dexamethasone alleviates traumatic proliferative vitreoretinopathy by inhibiting persistent inflammation and Müller cell gliosis in rabbits

AIM:To evaluate the effects of intravitreal slow-release dexamethasone on traumatic proliferative vitreoretinopathy(PVR)and Müller cell gliosis and preliminarily explored the possible inflammatory mechanism in a rabbit model induced by penetrating ocular trauma.METHODS:Traumatic PVR was induced in the right eyes of pigmented rabbits by performing an 8-mm circumferential scleral incision placed 2.5 mm behind the limbus,followed by treatment with a slow-release dexamethasone implant(Ozurdex)or sham injection.Left eyes were used as normal controls.The intraocular pressure(IOP)was monitored using an iCare tonometer.PVR severity was evaluated via anatomical and histopathological examinations every week for 6wk;specific inflammatory cytokine and proliferative marker levels were measured by quantitative real-time polymerase chain reaction,Western blot,protein chip analysis,or immunofluorescence staining.RESULTS:During the observation period,PVR severity gradually increased.Intense Müller cell gliosis was observed in the peripheral retina near the wound and in the whole retina of PVR group.Ozurdex significantly alleviated PVR development and Müller cell gliosis.Post-traumatic inflammation fluctuated and was persistent.The interleukin-1β(IL-1β)mRNA level was significantly upregulated,peaking on day 3 and increasing again on day 21 after injury.The expression of nod-like receptor family pyrin domain containing 3(NLRP3)showed a similar trend that began earlier than that of IL-1βexpression.Ozurdex suppressed the expression of IL-1β,NLRP3,and phosphorylated nuclear factor-kappa B(NF-κB).The average IOP after treatment was within normal limits.CONCLUSION:The present study demonstrates chronic and fluctuating inflammation in a traumatic PVR rabbit model over 6wk.Ozurdex treatment significantly inhibites inflammatory cytokines expression and Müller cell gliosis,and thus alleviates PVR severity.This study highlights the important role of IL-1β,and Ozurdex inhibites inflammation presumably via the NF-κB/NLRP3/IL-1βinflammatory axis.In summary,Ozurdex provides a potential therapeutic option for traumatic PVR.

Rabbit models of dry eye disease: comparative analysis

AIM:To report ocular changes in rabbits after the implementation of three different induction methods to create dry eye(DE)conditions and provides evidence of DErelated disease evolution.METHODS:Experimental methods were divided into 3 models.The first model used involved triple injection of complete Freund’s adjuvant,50µL each,also called the meibomian gland dysfunction(MGD)model.In the second model,DE conditions were created by the resection of nictitating membranes(NM),Harderian glands(HG),and main lacrimal glands(LG),also called the LGR model.The third model involved the topical administration of benzalkonium chloride(BAK)0.1%solution.The Schirmer test,ocular surface staining with fluorescein,and tear breakup time tests were implemented before and after excision.After euthanasia,the ocular tissues were dissected.Cornea,conjunctiva,and meibomian glands were treated with periodic acid–Schiff(PAS)staining and haematoxylin–eosin staining.RESULTS:The MGD model triggered inflammation of meibomian glands.It detected changes in the lipid layer of the tear film.The bilateral resection of NM,HG,and LG reduced the watering layer of the tear film.The topical administration of BAK of 0.1%solution impacted the mucosal layer of the tear film.CONCLUSION:Different changes are observed with different DE syndrome models.The composition of the tear film differ depending on which part of the eye is targeted.More studies need to be done to confirm whether an increased thickness of the cornea has any impact on the DE disease.

燕麦干草饲喂肉兔的营养价值评定

本试验旨在评定燕麦干草对肉兔的营养价值,为燕麦干草在肉兔配合饲料中的应用提供营养消化参数。试验分为2部分,燕麦干草营养成分分析和饲喂肉兔的消化参数测定。燕麦干草营养成分分析选择产自甘肃省的34个批次的燕麦干草样品进行,消化试验选择上述34个批次中代表性强(营养组成与测定均值差异较小)的燕麦干草。消化试验采用全收粪法进行:选取40只平均体重为(2.2±0.2)kg的70日龄新西兰白兔,随机分成2组,每组20只(公母各占1/2),代谢笼内单笼饲养。对照组饲喂基础饲粮,试验组饲喂试验饲粮(85%基础饲粮+15%燕麦干草)。预试期10 d,正试期7 d。结果显示:1)燕麦干草总能以及干物质、粗蛋白质、粗脂肪、粗灰分、钙、磷、无氮浸出物、粗纤维、中性洗涤纤维、酸性洗涤纤维和酸性洗涤木质素含量分别为15.33 MJ/kg、89.60%、9.71%、2.02%、12.54%、1.13%、0.21%、34.50%、30.83%、48.25%、38.87%和5.58%;肉兔对其表观消化率分别为42.51%、50.15%、56.58%、83.56%、25.12%、27.86%、20.36%、65.23%、28.87%、45.25%、30.87%和-5.86%。2)燕麦干草总氨基酸、赖氨酸、苏氨酸、蛋氨酸、精氨酸、半胱氨酸、组氨酸、缬氨酸、亮氨酸、异亮氨酸、苯丙氨酸和酪氨酸含量分别为8.67%、0.53%、0.45%、0.09%、0.49%、0.12%、0.22%、0.54%、0.09%、0.43%、0.46%和0.26%;肉兔对其表观消化率分别为58.23%、60.23%、62.52%、70.12%、68.25%、65.23%、58.23%、55.24%、62.35%、54.23%、59.68%和63.25%。综上可知,与苜蓿干草相比,燕麦干草粗蛋白质、粗脂肪含量偏低,粗纤维、中性洗涤纤维和酸性洗涤纤维含量较高,酸性洗涤木质素含量较低,粗灰分含量偏高,钙多磷少;肉兔对燕麦干草中各营养成分的消化率存在较大差异,其中总能、干物质、粗蛋白质、粗脂肪、无氮浸出物和限制性氨基酸消化率较高,纤维组分消化率一般,粗灰分、钙、磷和非限制性氨基酸消化率偏低。

Effect of taxifolin on cold-shock damages in spermatozoa in rabbits

Objective:To investigate the effect of taxifolin added to rabbit semen on freezing-induced cold-shock damages in spermatozoa.Methods:Semen was collected from six adult New Zealand rabbits once a week by artificial vagina.The collected semen was pooled at 38℃ and divided into four equal volumes.They were diluted with 0,50,100 and 200μM taxifolin-containing Tris+egg yolk extender at 38℃ and their temperatures were lowered to 4℃.Following equilibration,semen drawn into 0.25 mL straws were frozen in an automatic semen freezing device and stored in liquid nitrogen container at-196℃.Samples were thawed in 38℃ water for 25 s and the analyses of motility,kinematic parameters,morphological deformities,changes in membrane integrity,mitochondrial membrane potential,dead-live ratio,acrosomal damages and as well as oxidative stress analyses were performed in semen.Results:Addition of 50μM taxifolin significantly improved motility(total,progressive,rapid and static),high mitochondrial membrane potential and the ratios of spermatozoa with acrosomal damage compared to the control group.Compared to the control group,malondialdehyde(MDA)levels in the 50 and 100μM taxifolin groups were significantly lower,while the MDA level was high and viable spermatozoa ratio was low in the 200μM taxifolin group.There were no significant differences between the groups in terms of kinematic parameters,morphological deformities,membrane integrity and antioxidant levels.Conclusions:The low dose of taxifolin(50μM)has a positive effect and the high dose(200μM)has a negative effect.Therefore,it is concluded that the addition of low-dose(50μM)taxifolin to the extenders would be a useful additive in reducing cold-shock damage that occurs during freezing of rabbit semen.

A novel approach for 25-gauge transconjunctival sutureless vitrectomy to evaluate vitreous substitutes in rabbits

AIM:To improve the standard three-port vitrectomy for establishing and evaluating an endotamponade model in rabbits.METHODS:Three ports were prepared near the third eyelid of rabbits,and the infusion port was placed at the inferior nasal quadrant with the inserted cannula linking with a self-designed handheld rigid infusion catheter.All right eyes of rabbits underwent a modified 25-gauge vitrectomy and were subsequently filled with balanced salt solution,silicone oil,and eight-arm polyethylene glycols(8-arm PEGs)hydrogel separately for comparison.Ophthalmic examinations were performed regularly to record the changes after the surgery.RESULTS:Successful vitrectomy was achieved among 44 chinchilla rabbits.The mean operation time was 4.51±1.25min.Four eyes(9.1%)presented limited lens touch and two eyes(4.5%)showed retinal touch during surgery.Incision leakage was found in three eyes(6.8%)after surgery.There was no endophthalmitis,hemorrhage,or retinal detachment during the observation period and ophthalmic examinations after the implantation of vitreous substitutes.CONCLUSION:The modified technique of the standard vitrectomy applied in the endotamponade model in rabbits shows excellent safety and practicality.

Comment on:Optic neuropathy and increased retinal glial fibrillary acidic protein due to microbead-induced ocular hypertension in the rabbit

Dear Editor,W e read with great interest the study by Zhao et al[1],which evaluated optic neuropathy and changes in the amount of glial fibrillary acidic protein in the retina with intracameral injection of microbeads into rabbits.The objective was to investigate whether the injection of microbeads into the anterior chamber can induce chronic intraocular hypertension in a glaucoma model in rabbits.

Effect of a novel tyrosine kinase inhibitor nintedanib on bFGF and VEGF concentrations in a rabbit retinal vein occlusion model

AIM:To evaluate whether a novel tyrosine kinase inhibitor nintedanib could inhibit basic fibroblast growth factor(bFGF)and vascular endothelial growth factor(VEGF)simultaneously for retinal vascular disease in vivo.METHODS:After a laser induced rabbit retinal vein occlusion(RVO)model was made,0.5 mg of nintedanib was injected intravitreally in the left eye on the third day while the right eye was as a control.Intracameral samples were taken on the day before laser treatment and days 1,3,7,14,21,and 28 after treatment.Enzyme-linked immunosorbent assay(ELISA)was used to test the bFGF and VEGF-A concentrations in the aqueous humor.RESULTS:Both bFGF and VEGF-A rose significantly on the third day after laser treatment in both eyes.In the control eye the bFGF concentration peaked on the 14th day while the VEGF-A concentration dropped rapidly soon after the third day.After nintadanib injection in the study eye,both bFGF and VEGF-A showed a significant reduction on the 4th day(7th day after laser treatment)when compared to the control eye,and kept on low level in the following several weeks.CONCLUSION:Intravitreal injection of nintedanib can inhibit the expression of bFGF and VEGF in the process of RVO model to a certain extent,which is expected to become a new method for the treatment of retinal vascular diseases or fibrotic diseases.

Hypotensive Effects of an Aqueous Extract of Bambusa vulgaris (Poaceae) Leaves in Rabbit Oryctolagus cuniculus

Plant of Bambusa vulgaris is used in traditional medicine practice for the management of many pathologies including hypertension. The effect of the aqueous extract of the leaves of Bambusa vulgaris on blood pressure was evaluated in normotensive male rabbits. Blood pressure was measured by the invasive method. Thus, aqueous extract of Bambusa vulgaris leaves had been injected intravenously at doses ranging from 1 mg/kg to 30 mg/kg in rabbits anesthetized with thiopental. The effects of this extract on blood pressure were also evaluated in rabbits having previously received different doses of atropine. The aqueous extract of Bambusa vulgaris leaves induced a dose-dependent hypotension which is not canceled in the presence of atropine. The results obtained show that the extract has blood pressure lowering effect which may be mediated by muscarinic-type cholinergic receptors.

Rabbit Intensification Systems in Rwanda: Feeding Influence and Growth

A study was conducted in Northern Province of Rwanda, from the College of Agriculture and Veterinary Medicine, Busogo Campus located in Musanze district to evaluate the effect of feed type on rabbit growth in rabbit intensification systems in Rwanda. The Complete Randomized Design (CRD) was used and data were collected on rabbit growth weekly for a period of 12 weeks. The experiment was composed of three treatments replicated ten times. The treatments included three types of feeds namely;cabbage combined with Mucuna pruriens added to local forage (I), cabbages combined with Leucaena leucocephala added to local forage (II) and a control composed of other varieties of locally available forage, such as Bidens pilosa, Crassocephalum vitellium and Galinsoga parviflora (III) which was considered as the control (Farmers practice). The feeds were given to ten rabbits separated in individual cages, and each rabbit was considered a replicate. Water was given ad libitum. One month old rabbits (weaners) were used and data were collected after one week of adaptation for 12 weeks. The results showed that the mean of weight gain after 12 weeks was 783.3 g, 760.7 g and 705.7 g for feed type I, II and III respectively. The difference between means of feed types after 12 weeks was not significant (p > 0.5), which implied that rabbit growth did not depend on the feed type. The mean weight gain after 8 weeks was 707.5 g, 661.4 g and 577.1 g for feed type I, II and III respectively. At 8 weeks, the difference between means of feed types was significant (p Mucuna pruriens combined with cabbage and local forage were growing faster than rabbits from other treatments at 8 weeks. The researchers recommended that farmers should be facilitated with feeding materials by the concerned institutions. Training of rabbit farmers and further researches on locally available feeding materials were also given as recommendations at the end of this study.

Analysis and Identification of miRNA Expression in the Skeletal Muscle of Sichuan White Rabbits

[Objectives]miRNAs play an important role in the proliferation and differentiation of different myoblasts.This study was conducted to elucidate the complex genetic mechanisms that affect the meat production performance of Sichuan white rabbits and reveal the regulatory role of miRNAs in their muscle growth and meat quality formation.[Methods]Three constructed skeletal muscle libraries of Sichuan white rabbits aged six months were sequenced by the solexa technology to identify known miRNAs,predict new miRNAs and construct an expression profile of muscle miRNAs.[Results]A total of 511 known miRNAs and 42 miRNAs were detected in 34089472 pure sequences,and the proportion of miRNAs with a length of 22 nt was the highest.The number of known miRNA sequences accounted for 71.38%of pure sequences,which was much higher than the proportion of other types of RNAs.The proportion of sequences from exons was 0.38%,indicating a low degree of mRNA degradation in the samples.Base U had the highest proportion at the first position,and the bases with the highest proportions at positions 8 and 10 were U and A,respectively.Muscle-specific miRNAs(miR-1,miR-133,and miR-206)ranked in the top 10 in terms of expression level.The number and expression levels of new miRNAs were lower than those of known miRNAs.The length distribution,base bias at different positions and expression profile characteristics of miRNAs might be related to the biological function of miRNAs in regulating muscle proliferation and differentiation and the action mechanisms with target genes.[Conclusions]The identification and expression of miRNAs in muscle tissues of Sichuan white rabbits will help to understand the complex molecular mechanisms of meat production performance and provide a theoretical basis for the functional research of miRNAs in meat rabbits.

饲粮中葛藤不同添加量对肉兔养分表观消化率、胃肠道发育及消化酶活性的影响

为探讨葛藤在肉兔饲粮中的应用效果,开展葛藤的常规营养成分分析,选取35±2日龄体重(0.88±0.13 kg)相近的断奶伊拉兔肉兔200只,随机分成4组,每组5个重复,每个重复10只,分别饲喂添加0(对照组)、15%、25%、35%葛藤的试验饲粮。预试期7 d,正试期42 d。于正试期第36天开始进行为期7 d的消化代谢试验,分析肉兔养分表观消化率,试验结束后测定分析肉兔盲肠消化酶活性及胃肠道的变化。结果显示,饲粮中添加不同量的葛藤:1)提高了各处理组肉兔对饲粮中干物质、粗蛋白、粗纤维、中性洗涤纤维、酸性洗涤纤维、酸性木质素、粗脂肪和半乳糖醛酸的表观消化率,35%葛藤组表观消化率最高,显著高于15%葛藤组与对照组(P<0.05)。2)提高了肉兔盲肠中纤维素酶、半纤维素酶与果胶酶活性,15%葛藤组酶活性最高,显著高于对照组(P<0.05)。3)对肉兔胃及小肠的相对长度无显著影响(P>0.05),但显著提高了盲肠相对长度和重量、胃相对重量与小肠相对重量,25%与35%葛藤组显著高于对照组(P<0.05)。4)提高了肉兔空肠、回肠与十二指肠的绒毛长度、绒毛长度/隐窝深度比值(V/C)和黏膜层厚度,降低了隐窝深度值,35%葛藤组肉兔回肠、十二指肠的绒毛长度显著高于对照组(P<0.05),空肠、回肠的隐窝深度低于对照组,空肠、回肠的V/C显著高于对照组(P<0.05)。因此,生长肉兔饲粮中最适宜的葛藤添加量为35%。

盐酸青藤碱诱导黏连性膝关节强直家兔成纤维细胞凋亡的机制研究

目的观察盐酸青藤碱对膝关节黏连强直的家兔成纤维细胞增殖和相关基因表达的影响,并进一步尝试探讨其对抗膝关节黏连强直的作用机制。方法以体外培养法培养成纤维细胞,并设对照组、盐酸青藤碱低中高浓度实验组。CCK-8法检测成纤维细胞增殖的情况;实时荧光定量聚合酶链反应(real-time quantitative PCR,RT-qPCR)法检测经过盐酸青藤碱处理后,成纤维细胞相关基因mRNA表达的改变,用ELISA法检测药物的作用对血清中炎症因子等水平的影响,蛋白质印迹法(Western blot)检测相关蛋白质的表达。结果盐酸青藤碱能降低成纤维细胞存活率,且随浓度升高存活率逐渐降低。盐酸青藤碱中各个组的效果均十分明显(P<0.05)。在相关基因的mRNA表达层面,与对照组比较,盐酸青藤碱各组炎症因子均显著下调(P<0.05),凋亡蛋白的表达量显著上升、Bcl-2的mRNA表达量下降(P<0.05),而PI3K/mTOR/AKT3信号通路分子的mRNA表达量均下降(P<0.05)。在蛋白质表达层面,与对照组相比较,中、高剂量盐酸青藤碱组血清中炎症因子IL-6、IL-8、IL-1β、TGF-β的水平均明显下调(P<0.05),凋亡蛋白cleaved-PARP、cleaved caspase-3/7及Bax的表达量均上调,并且与给药剂量成正相关,而抗凋亡蛋白Bcl-2、PI3K/AKT3/mTOR信号通路的表达量则与给药剂量成负相关。盐酸青藤碱对家兔膝关节成纤维细胞的存活表现为显著的抑制作用,作用机制或与下调炎症因子IL-6、IL-8、IL-1β的表达,并促进凋亡蛋白cleaved-PARP、cleaved caspase-3/7及Bax的表达,抑制Bcl-2的表达,抑制其下游PI3K/AKT3/mTOR信号通路的基因表达有关。结论盐酸青藤碱可抑制黏连性膝关节强直家兔膝关节成纤维细胞的炎症反应和加速成纤维细胞的凋亡,或可通过该机制为改善和治疗黏连性膝关节强直提供新的方法。