表达大肠杆菌K88ac-ST1-LTB融合蛋白工程菌株的免疫原性研究

已构建的能表达大肠杆菌K88ac-ST1-LTB融合蛋白的工程菌株BL21（DE3）（pXKST3LT5)及其表达产物经动物试验证实没有毒性反应。用从IPTG诱导的工程菌中提取的包涵体或经甲醛灭活的工程菌制成抗原，免疫小鼠，结果免疫小鼠至少能抵抗2MLD的大肠杆菌强毒株C83902（K88ac,ST^ ,L^ )的攻击，用提取的包涵体免疫家兔后，采集的血清能够中和天然ST1的毒性，这表明构建的工程菌株BL21（DE3）（pXKST3LT5)可以作为预防幼畜大肠杆菌性腹泻基因工程菌苗的候选株。     

Delivery of growth factors using gene therapy to enhance bone healing

OBJECTIVE: To review the delivery of growth factors using gene therapy for enhancing long-bone fracture healing. STUDY DESIGN: Literature review. METHODS: MEDLINE and CAB Abstracts literature search (1980-2004). RESULTS: Non-union and infected non-union are relatively common complications of long-bone fractures in human and veterinary patients. Growth factors are cytokines that regulate many cell functions important in fracture healing. Exogenous growth factors can be delivered to the fracture site as recombinant proteins or using gene therapy. Recombinant human bone morphogenetic protein-2 and -7 (rhBMP-2 and -7), in particular, enhance fracture healing in numerous experimental and clinical studies. Some limitations with use of recombinant proteins may be overcome by use of gene therapy. Gene therapy involves delivery of the growth factor gene to cells at the fracture site using a viral or non-viral vector. The gene is then expressed (protein synthesis) by cells at the fracture site. Delivery of the BMP gene to the fracture site using gene therapy has been evaluated in laboratory animal models of non-union, with favorable results and without complications. CONCLUSION: Delivery of growth factors, particularly members of BMP family, to the fracture site using gene therapy may be a method to enhance fracture healing. Use of this technology may improve the prognosis for patients with long-bone fractures. CLINICAL RELEVANCE: Clinical application of gene therapy could improve the prognosis for human and veterinary patients with long-bone fractures, but has not been evaluated clinically.     

喷雾干燥鱼浆蛋白粉营养成分分析

本试验对鱼粉加工废水鱼浆蛋白粉(经浓缩、水解、喷雾干燥得到的副产品)进行营养成分分析,其中粗蛋白质含量达64.08%,钙2.67%、磷1.12%,钙磷比例合理;各氨基酸含量分别为:赖氨酸3.29%,蛋氨酸0.94%,谷氨酸5.17%,牛磺酸2.50%。     

Analytical and Clinical Validation of a Time-resolved Immunofluorometric Assay (TR-IFMA) for Canine C-reactive Protein in Serum

A time-resolved immunofluorometric assay (TR-IFMA) was developed for the determination of C-reactive protein (CRP) in canine serum. CRP was isolated from canine acute-phase serum by affinity chromatography on agarose coupled with phosphorylethanolamine. This isolated dog CRP was used as standard to calibrate the assay. Intra-assay and inter-assay coefficients of variation were in the ranges 5.3–7.1% and 4.8–13.3%, respectively. Accuracy, evaluated by adding 2 and 10 μg/ml of CRP to serum samples, provided recoveries of 99.9% and 106.8%. High correlation was found between CRP measurements by TR-IFMA and a by commercial enzyme-linked immunosorbent assay (R² = 0.98). The limit of detection for the TR-IFMA method was 0.000067 μg/ml and the measurement of CRP in serial dilutions of acute-phase dog sera generated curves with the same slope as the one constructed with purified CRP. The TR-IFMA provides a precise, accurate and highly sensitive assay for CRP determination in dog samples. CRP levels in dogs with different diseases ranged between 10.2 and 210.7 μg/ml and were significantly higher than those observed in healthy dogs (< 7.1 μmg/ml).     

马链球菌兽疫亚种类M蛋白亚单位疫苗小鼠免疫试验

为获得控制猪链球菌病安全高效的疫苗，本试验进行了马链球菌兽疫亚种类M蛋白亚单位疫苗的研制。应用热酸法提取马链球菌兽疫亚种ATCC35246株类M蛋白，并分别采用羟基磷灰石（HAT）层析和冷酒精沉淀法进行纯化，结果得到较纯的类M蛋白。用热酸提取的粗制类M蛋白、2种纯化的类M蛋白及全菌苗免疫小鼠，结果保护率分别为92％（11／12）、100％（12／12）、100％（12／12）和83％（10／12），表明类M蛋白亚单位疫苗有进一步开发的必要。     

Relationship between latency of brainstem auditory‐evoked potentials and head size in dogs

Summary

Cranium and brainstem dimensions were measured in 32 postmortem dog heads. Positive correlations were found between cranium length (CL) and brainstem length (BL) (r=0.87), between cranium width (CW) and brainstem width (BW) (r=0.83), and between cranium distance (CD = CL CW/2) and brainstem distance (BD = BL+BW/2) (r=0.91). Positive correlation coefficients were also found between CL and CW (r=0.90), and between BL and BW (r=0.85). It was concluded that head size accurately reflected brainstem size. A least squares estimation of the brainstem distance (BD) from CL and CW values was BD = 10.9 + 0.16 (CL CW/2) (BD, CL and CW in mm).

Brainstem auditory evoked potentials (BAEPs) and cranium dimensions were measured in 43 dogs (86 ears) with different head size, body size, sex and age. Wave form, absolute and interpeak latencies and correlation coefficients, relating latencies to cranium dimensions and body weight, were analysed CL, CW, and CD were positively correlated with body weight (r=0.93, 0.70 and 0.93, respectively), and CL, CW, and CD were correlated with age (r=0.33, 0.52 and 0.40, respectively). BAEPs consisted of five distinct positive peaks (I to V). Secondary positive peaks following peaks I and II were seen in 60% (I') and 90% (II') of the recordings. Late waves were recorded in 90% (VI), 50% (VII), and 25% (VIII) of the recordings. Latencies increased with decreasing stimulus intensity level (from 90 dB to 10 dB hearing level, HL),especially for peaks I, II, V, and the I‐V interpeak interval Absolute and interpeak latencies were positively correlated with cranium distance and body weight. Correlation coefficients increased as wave latencies increased At 90 dB HL, the highest correlation coefficients, relating cranium distance to peak V and the I‐V interpeak latency, were 0.55 and 0.53 (P < 0.00001), respectively. Regression analysis showed that each 1 cm increase in cranium distance was accompanied by an increase of 0.006 ms in the latency of wave I, 0.03 ms for wave III, 0.05 ms for wave V, and 0.05 ms for the I‐V interpeak interval Regression analysis showed that an increase of 1 kg in body weight was accompanied by an increase of 0.001 ms in the latency of wave I, 0.005 ms for wave III, 0.011 ms for wave V, and 0.01 ms for the I‐V interpeak interval. It is concluded that head size, which accurately reflects brain size, is a relevant source (25%) of intersubject variance of BAEP latencies in the dog.     

内质网应激介导的胰岛β细胞凋亡研究进展

胰岛β细胞具有十分发达的内质网(ER),并且对应激非常敏感。多种原因都会使ER的稳态被破坏,引起未折叠蛋白反应(UPR)以及Ca^2+稳态失衡,进一步诱发内质网应激(ERS)。适度的ERS有利于恢复其稳态,过度的ERS会导致ER功能受损,并进一步诱导胰岛β细胞凋亡,从而介导糖尿病的发生、发展。因此,对生理及病理条件引发的ERS进行分析,探讨ERS介导的胰岛β细胞凋亡,可为糖尿病的深入研究奠定理论基础。     

饲料资源量概念及其判断

首先提出并讨论了饲料资源量的主体部分就是饲料能量总量及饲料蛋白质总量的问题。然后,用相关及回归方法筛选出了牛代谢能及粗蛋白质指标。在此基础上,建立了对饲料资源的品质和数量进行判断的蛋能结构指数及蛋能丰度指数体系。