Optimization and performance validation for the fully automatic blood type analysis system

To evaluate and validate the application of fully automatic blood type analysis system parameters under actual lab conditions. All key system parameters were optimized and validated accordingly. The optimized parameters were centrifugal speed at 550 rpm; centrifugal time 20 min; resuspension speed 1,200 rpm; resuspension time: 45 s; incubation temperature 25℃ ; incubation time 400 s; incubation rate: 0 rpm. The sampled red blood cell concentration was 3%. The ratio of plasma to red blood cells reagent was 60 μL:30 μL; the ratio of antibody (reagent) to sampled diluted red blood cell was 30 μL:30 μL. After applying our key parameters for optimization and validation, the automatic blood type detection system's performance was found to meet the relevant requirements, effectively improving the accuracy and reliability of the detection system.     

Effects of the usage of dried brewing yeast in the diets on the performance, egg traits and blood parameters in quails

This experiment was carried out to determine the effects of the usage of dried brewing yeast in quail diets on laying performance, egg traits and blood parameters. A total of 240 Japanese quails (Coturnix coturnix japonica) aged 10 weeks were randomly allocated into one control group and three treatment groups. Each group was divided into five replicates as subgroups, comprising 12 quails each. Dried brewing yeast (Saccharomyces cerevisiae) was used at the levels of 1.5%, 3.0% and 4.5% in the diets of the first, second and third treatment groups, respectively. Soyabean meal was replaced with dried brewing yeast. The diets were formulated to be isocaloric and isonitrogenous. The experimental period lasted 18 weeks. Dietary treatments did not significantly affect body weight, daily feed intake, daily protein intake, egg production, egg weight, feed efficiency, mortality, egg shell thickness, egg albumen index, egg yolk index, egg Haugh unit, the percentages of egg shell, albumen and yolk, excreta moisture and small intestinal pH. Inclusion of 3% and 4.5% dried brewing yeast in diets reduced egg yolk cholesterol concentration as mg per yolk and mg per g yolk (P < 0.01). Blood serum cholesterol of groups fed diets with dried brewing yeast was significantly lower (P < 0.01) than that of the control group. Feeding diets containing 3.0% and 4.5% dried brewing yeast resulted in significant increases (P < 0.01) in blood serum levels of total protein, alanine aminotransferase at the end of the experiment. Blood serum levels of uric acid, triglyceride, aspartate aminotransferase and alkaline phosphatase were not affected by dietary dried brewing yeast. It is concluded that dried brewing yeast can be used up to 4.5% in the diets of laying quails without adverse effects on the measured parameters.     

Effects of glutamine on growth performance,nutrient content,fatty acid profile,and blood parameters of rainbow trout (Oncorhynchus mykiss)

In this study, different amounts of glutamine were added to the diet of rainbow trout, and they were then fed for a period of 90 days. The current research investigated the effects of glutamine on various aspects of rainbow trout, including growth performance, condition factor, viscerosomatic index, hepatosomatic index, carcass composition, fatty acid profile, hematological parameters, and biochemical parameters. The study's findings revealed that adding glutamine to the diet of rainbow trout had a beneficial impact on their growth features. The rainbow trout group that was fed a 2% concentration of glutamine experienced the most notable increase in growth rate. A statistically significant difference in growth was observed among all groups (p < 0.05). Adding glutamine to the diet increased the amount of protein and decreased the fat content in the flesh of the fish. Glutamine exerted an influence on the blood and biochemistry parameters of fish, as well as their fatty acid composition. In conclusion, the inclusion of glutamine in the diet of fish could potentially enhance their immune system, improve the quality of their muscles, and enhance their growth performance.     

饲粮中添加杜仲叶对断奶仔猪生长性能、抗氧化力和免疫功能的影响

在断奶仔猪饲粮中添加不同形式的杜仲叶,研究其对仔猪生长性能、腹泻率、抗氧化力及免疫功能的影响,以评估它们在养殖初期替代抗生素的能力。将断奶仔猪(杜×长×大)随机分成对照组、抗生素组(75 mg/kg金霉素)和三个杜仲叶组(杜仲叶提取物、发酵杜仲叶、杜仲叶干粉),每组10头,进行28天养殖试验。结果表明:与对照相比,杜仲叶提取物组仔猪平均日增重(ADG)显著提高并降低料重比(F/G)(P<0.05);与抗生素组相比,杜仲叶提取物组和杜仲叶发酵组ADG和F/G均无显著差异(P>0.05);各组之间平均日采食量(ADFI)差异不显著(P>0.05)。与对照组相比,抗生素组和杜仲叶提取物组仔猪腹泻率显著降低(P<0.05)。与对照组相比,抗生素组和三种杜仲叶组均降低仔猪血液丙二醛含量和提高总抗氧化力(P<0.01),而抗生素组、杜仲叶提取物组和杜仲叶发酵组还显著提高血液免疫球蛋白IgM含量(P<0.01),且表现出提高血液IgG含量的趋势(P=0.06)。与其他各组相比,杜仲叶提取物组可显著提高肝脏指数(P<0.05);肝脏重量、脾脏重量和脾脏指数在各组之间无显著差异(P>0.05)。饲粮中添加杜仲叶提取物可改善断奶仔猪的生长性能,降低腹泻率,提高仔猪抗氧化力和免疫功能,其效果优于发酵杜仲叶和杜仲叶干粉,并具备一定的抗生素替代潜力。     

高压蒸汽灭菌柜性能验证方法的研究

高压蒸汽灭菌柜在使用之前和运行一定时间后,必须进行性能验证,采用热分布,热穿透和微生物挑战试验法对GE,GEV型脉冲式蒸汽灭菌柜的性能进行验证,多孔物质及流体物质灭菌循环中,灭菌腔内不存在冷点,具备有效的热穿透力,嗜热脂肪芽孢杆菌（ATCC7953）在规定灭菌时间内被完全杀死。因而确认高压灭菌柜的各项性能均达到生产要求,由此建立的一套验证方案及试验方法和结果得到了国家GMP认证中心的认可。     

基于氨基酸特征序列对人类Rh血型系统的蛋白质结构分析

利用代数学中同态思想和物理中的“粗粒化”思想,以及HP模型,根据a,t,c,g的化学结构分类,提出了DNA序列的特征序列概念（σ-,τ-,σ∩τ-）并推广到蛋白质序列中,从而给出一种数值刻划,将蛋白质序列简化成一个（0,1）序列,基于上述给出特征序列的方法,根据氨基酸分子量与简并度的关系,提出了另外一种DNA序列的特征序列概念（-）并推广到蛋白质序列中,进而给出了另外一种数值刻划,将蛋白质序列简化成一个（0,1,2）序列,通过比较RHD基因和RHCE基因的特征序列的数值刻划图,得出RHD基因和RHCE基因均偏爱使用低分子量且高简并度的氨基酸。     

Influence of probiotic supplementation on blood parameters and growth performance in broiler chickens

Effects of commercial probiotic (Bactocell®) on growth performance and blood parameters were evaluated. A total of 800 one day-old Ross broiler chicks were raised over 42 days. Chicks were wing-banded, weighed individually and randomly allocated into four equally major groups each having two replicates. Chicks of group 1 (control group) were fed the starter and finisher diets that did not supplemented with probiotic. The chicks of groups 2, 3, and 4 were fed the control starter and finisher diets supplemented with 1.6 g, 1 g and 0.8 g of probiotic per kg feed, respectively. Weekly body weight, feed consumption and feed conversion were measured. Blood parameters at 1, 4 and 6 weeks of age including packed cell volume (PCV), haemoglobin (Hb), total protein, albumin, total lipid and cholesterol were determined. All birds were kept under similar environmental, managerial and hygienic conditions. The results of the current study revealed that there was no significant change for Hb and PCV concentrations among different groups at all studied times. Also, total protein, lipids and albumin concentrations were not affected by probiotic supplementation. Chicken fed a diet containing various levels of probiotic showed a significant decrease (p ⩽ 0.05) in cholesterol concentration compared to control group. Probiotic supplementation significantly increased the body weight and daily weight gain of broiler chicks at late ages (3–6 weeks). Also, the birds fed on probiotic levels 1 and 0.8 g/kg diet exhibited higher body weight among chicken groups at 6 weeks of age. Improved feed conversion was noticed in birds fed a diet supplemented with probiotic. There was no significant difference in mortality rate among groups. We concluded that use of selected commercial probiotic resulted in improved performance parameters and reduced serum cholesterol in broiler chickens. Moreover, supplementation of the probiotic to broilers in the levels of 1 and 0.8 g/kg diet was found to be better than control and 1.6 g/kg level indicating that increasing dietary probiotic level does not has the best performance.     

Optimization of the genotyping‐by‐sequencing strategy for population genomic analysis in conifers

Flexibility and low cost make genotyping‐by‐sequencing (GBS) an ideal tool for population genomic studies of nonmodel species. However, to utilize the potential of the method fully, many parameters affecting library quality and single nucleotide polymorphism (SNP) discovery require optimization, especially for conifer genomes with a high repetitive DNA content. In this study, we explored strategies for effective GBS analysis in pine species. We constructed GBS libraries using HpaII, PstI and EcoRI‐MseI digestions with different multiplexing levels and examined the effect of restriction enzymes on library complexity and the impact of sequencing depth and size selection of restriction fragments on sequence coverage bias. We tested and compared UNEAK, Stacks and GATK pipelines for the GBS data, and then developed a reference‐free SNP calling strategy for haploid pine genomes. Our GBS procedure proved to be effective in SNP discovery, producing 7000–11 000 and 14 751 SNPs within and among three pine species, respectively, from a PstI library. This investigation provides guidance for the design and analysis of GBS experiments, particularly for organisms for which genomic information is lacking.     

The AB blood group system in wild felids

The blood type of 131 non-domesticated felids belonging to 26 felid species was surveyed in this study. Based upon a tube hemagglutination assay established for domestic cats, 80% of felids had type-A, 18% type-B, and 2% type-AB blood. Felids in the Puma group and African and Asian golden cats had blood type B, whereas all other species were found to have blood type A. Two cheetahs and one bobcat had type-AB blood. Red cell glycolipids analysed by high performance thin layer chromatography revealed a similar ganglioside pattern in wild cats as reported in domestic cats. Independent of the AB blood group system, incompatible blood crossmatch reactions were detected between different felid groups. In conclusion, wild felids display the same AB-erythrocyte antigens as domestic cats, and the same blood typing procedures can be applied for wild and domestic felids.     

单克隆抗体血型定型试剂转瓶培养生产工艺

研制成功单克隆抗体ABO血型定型试剂转瓶培养生产工艺,生产周期从静置培养生产的5～6天缩短到2～3天。产品所有质量指标均达到或超过国家标准,特别是抗体效价明显提高。实验及生产结果显示,该法简单易行,经济快速．投入低,产出高,生产时间短,产品质量高。     

以人类血型为遗传学案例教学的思考与实践

血型是人类日常生活中非常常见的一种遗传表型, 拥有丰富的遗传学内涵。随着科技的发展, 其内涵不断得到新的揭示, 新的研究结果不断补充, 持续吸引着人们对血型遗传机制的探索。血型遗传案例除了与孟德尔遗传和连锁遗传、基因突变和染色体畸变四大内容关联外, 还涉及到其他多方面的遗传学知识点。在教学中, 依据遗传学的知识脉络, 贯穿以ABO血型作为经典案例, 结合拓展的白细胞血型, 孟买、Rh、MN等血型的遗传规律及其应用, 并且开展相关的实验教学, 理论联系实际, 增强了学生的兴趣, 提高了教学效果。在遗传学实验教学中, 有80%的学生选择ABO血型鉴定这个自选实验, 并表示出对这个实验的浓厚兴趣。在讲授相关知识点时, 用恰当的血型案例为引导, 设计相关的讨论主题, 开展PPT展示性讨论和辩论式讨论, 所有的学生都积极主动参与进来, 与现实生活相结合, 引导学生思考问题, 使学生的思维在辨析中得到操练, 提高分析问题和解决问题的能力, 深刻理解遗传学基本理论知识。     

ABO血型综合实验基因分型技术简化及其群体遗传分析拓展

ABO血型是生活中最常见、运用最广泛的遗传性状之一。人类ABO血型由I ^A、I ^B和i 3个复等位基因决定,它们负责编码不同的糖基转移酶,进而决定3种血红细胞表面抗原。ABO血型涉及复等位基因、基因互作、单核苷酸多态(SNP)、基因演化等多个关键知识内容,是理想的遗传学教学案例。本文以ABO血型为研究对象,对遗传学实验进行了创新与整合。首先,在分子遗传学模块中建立了新颖的ABO血型基因分型方法：基于SNP位点设计特异性引物,通过实时定量PCR鉴定基因型; 其次,在群体遗传学模块中创新了基因演化的实验教学方法,开发群体遗传学软件,利用计算机模拟不同条件下ABO血型决定基因频率的演化趋势。这些教学改革举措旨在丰富遗传学实验内容,拓展教学手段,提高学习效率。     

Frequencies of ABO, Rh and Kell phenotypes in couples from Han, Kazak, Uyghur and Hui in Xinjiang: an inheritance simulation model for blood group incompatibility in new-born

The Xinjiang region with residents from more than 13 minorities represents an area of many diverse ethnicities. This ethnic diversity in relation to their blood groups and immune status may have a consequential impact on the clinical status of married couples. To evaluate the risks of haemolytic disease in new-born infants, we investigated the rate of blood-group incompatibility among 487 married couples from four ethnic minorities, namely the Han, Hui, Uyghur and Kazak populations. Han minority married couples showed significantly different ABO, Rh and K phenotype frequencies between marrial relationship, whereas there was no significant difference in ABO, Rh and K phenotypes between the Uyghur, Hui and Kazak .There was a significant difference between ABO blood types in Han married couples, in the Kazak Rh-C phenotype and in the Uyghur Rh-D phenotype. The Hui married couples only demonstrated ABO, Rh and K phenotypes. The Hui minority showed the highest incompatibility rate for Rh-C and Rh-E phenotypes between mothers and their new-born infants. The highest incompatibility rate for the ABO phenotype occurred in the Kazak group. These results particularly demonstrate the clinical issues relating to ABO and Rh incompatibility, in the Kazak and Hui minorities, respectively.     

人类血型性状综合遗传大实验的设计与教学实践

综合大实验能够训练学生灵活应用理论知识并掌握实验技能,成为当前实验课教学改革的重要方式。本文以人类的ABO血型性状为实验对象,设计了“人类ABO血型分子基因分型与群体遗传平衡分析”大实验。实验中提取同学唾液中黏膜细胞的DNA,经过PCR扩增目的片段、酶切及电泳分离一系列分子遗传技术分析,鉴定出每位同学的基因型;然后以全班同学为一个类似孟德尔群体调查ABO血型的各种基因型频数,用Popgene软件分析各种群体遗传参数。通过开放教学不仅让学生掌握了分子遗传实验技术和群体遗传分析技术及软件应用,还让学生自主设计方案优化分子技术环节,提高学生驾驭知识的能力。通过5年的教学探索与实践,建立了稳定的分子遗传实验体系,能够清楚地检测出ABO血型的6种基因型：I^AI^A、I^Ai、I^BI^B、I^Bi、I^AI^B、ii;综合了分子遗传与群体遗传的实验教学,统计全班同学6种基因型的频数,直接计算3个复等位基因的频率,进而应用软件分析群体遗传各种参数;实现了学生自主设计并完成实验的开放式实验教学;大实验教学获得了学生的好评,取得了很好的教学效果。该大实验可直接应用于生物类专业的遗传学实验教学,其中的教学理念和方法还可推广应用于其他生物学实验教学。     

Cell type specific DNA methylation in cord blood: A 450K-reference data set and cell count-based validation of estimated cell type composition

Epigenome-wide association studies of prenatal exposure to different environmental factors are becoming increasingly common. These studies are usually performed in umbilical cord blood. Since blood comprises multiple cell types with specific DNA methylation patterns, confounding caused by cellular heterogeneity is a major concern. This can be adjusted for using reference data consisting of DNA methylation signatures in cell types isolated from blood. However, the most commonly used reference data set is based on blood samples from adult males and is not representative of the cell type composition in neonatal cord blood. The aim of this study was to generate a reference data set from cord blood to enable correct adjustment of the cell type composition in samples collected at birth. The purity of the isolated cell types was very high for all samples (>97.1%), and clustering analyses showed distinct grouping of the cell types according to hematopoietic lineage. We explored whether this cord blood and the adult peripheral blood reference data sets impact the estimation of cell type composition in cord blood samples from an independent birth cohort (MoBa, n = 1092). This revealed significant differences for all cell types. Importantly, comparison of the cell type estimates against matched cell counts both in the cord blood reference samples (n = 11) and in another independent birth cohort (Generation R, n = 195), demonstrated moderate to high correlation of the data. This is the first cord blood reference data set with a comprehensive examination of the downstream application of the data through validation of estimated cell types against matched cell counts.     

Productive performance,fertility and hatchability,blood indices and gut microbial load in laying quails as affected by two types of probiotic bacteria

This study investigated two kinds of probiotic bacteria (Bacillus toyonensis, B1 and Bifidobacterium bifidum, B2) on laying Japanese quail’s performance, egg quality, fertility and hatchability, blood biochemical characteristics and microbiological parameters. A total of 270 mature quails (180 females and 90 males) were distributed into ten groups in a completely randomized design at eight weeks of age. The experimental groups were as follows: T1: basal diet only (control); T2-T5, basal diet plus 0.05, 0.075, 0.10 and 0.125% B1, respectively; T6: basal diet plus 0.10% B2; T7-T10: basal diet plus 0.05, 0.075, 0.10 and 0.125% B1 plus 0.05% B2, respectively. Results revealed that egg number (EN) and egg weight (EW) were gradually increased (P < 0.01) as the levels of both probiotic types increased. The feed conversion ratio (FCR) was significantly (P < 0.05) better within the total experimental period (8–20 weeks) due to B1 alone or/with B2 supplementation. Values of yolk percentage (Y%) were statistically (P < 0.01) higher only at 8–20 weeks of age and T10 recorded the highest value. By increasing the level of probiotics, fertility and hatchability percentages (F% and H%) were gradually increased (P < 0.01 and P < 0.05). Creatinine (CR) level was statistically reduced in birds fed T4 diet. Also, urea-N and aspartate aminotransferase (AST) levels were reduced in treated birds. The opposite was found regarding alkaline phosphatase (ALP). Conclusively, using B1 and B2 enhanced the productive performance, some egg quality traits, fertility and hatchability, digestive enzyme activities, and reduced the harmful bacteria in the gut of laying Japanese quail. Our findings could recommend to apply T4 (basal diet + 0.10 % B1), T6 (basal diet + 0.10% B2) and T9 (basal diet + 0.10% B1 + 0.05% B2) levels for the best results.     

Morphometric requirements for image analysis and the IBAS interactive automatic image analysis system

The morphometric demands made on an image analysis system are discussed, as is the IBAS interactive automatic system, which meets those requirements. The modular design of the IBAS image analysis system, with its tailor-made processors for image processing, system control and pattern recognition, gives speed and flexibility. The IMAGE image analysis language guarantees user friendliness, and, last but not least, the enormous amount of software offers accurate, reproducible measurements and dedicated evaluation programs.     

Optimization,validation, and identification of two reliable antibodies for immunodetection of WNT5A

WNT5A is a secreted, noncanonical WNT signaling protein that has been reported to promote progression of several types of cancer, including oral squamous cell carcinoma. Many WNT5A antibodies are available commercially for immunohistochemistry (IHC) and western blot analysis. Validation of the primary antibodies, however, is often neglected. We characterized antibodies for detecting WNT5A by IHC and western blot analysis. We evaluated one polyclonal and three monoclonal commercially available WNT5A antibodies. After optimization of the IHC assay, all four antibodies showed cytoplasmic WNT5A expression in tissue samples; in contrast, only one antibody detected WNT5A in western blots. A pre-absorption test with recombinant WNT5A showed that AF645 and 3A4 antibodies specifically detected WNT5A in different assays. We suggest that the monoclonal 3A4 antibody is the most appropriate for use with IHC, while the polyclonal AF645 antibody is the best for western blot analysis.