应用间接血凝试验检测牛肺疫抗体

自制1.5％绵羊红细胞诊断液并用于检测牛肺疫抗体,结果如下:(一)三批标准阳性血清(批号为8601、8301、8001)凝集价均在160倍以上,标准阴性血清(批号为8601)凝集度仅为10倍。15份被检血清IHA的阳性检出率(8/15)高于补体结合反应(CF,6/15)且凝集价均在40倍以上。(二)用牛肝片吸虫病、布氏杆菌病和附红细胞体病阳性血清作类症鉴别诊断,凝集度仅为10倍。(三)把牛肺疫CF8502抗原作倍比稀释,加2个凝集单位牛肺疫阳性血清8601和8301进行间接血凝抑制试验,分别被32倍和16倍稀释的抗原所抑制,本项试验结果表明IHA用于检测牛肺疫,具有检出率高、特异性强、设备要求简单、容易操作等特点,适于在现地推广应用。     

Protective effect of recombinant SCR15-18 domain of human complement receptor type 1 on intestinal ischemia and reperfusion injury

AIM: To investigate the protective effect of recombinant SCR15-18 domain of human complement receptor type 1 (CR1-SCR-15-18) on intestinal ischemia and reperfusion in a rat model. METHODS: Sprague-Dawley rats were randomly divided into 3 groups: sham operation(SO) group, ischemia and reperfusion (I/R) group and CR1-SCR15-18 treatment group. The superior mesenteric artery of the rats was clamped for 30 min followed by 60 min of reperfusion. PBS alone or CR1-SCR15-18 protein (30 mg/kg) in PBS was intravenously administered 5 min before reperfusion. Intestinal vascular permeability, myeloperoxidase (MPO), malondialdehyde (MDA) and superoxide dismutase (SOD) were measured. The histopathological changes of intestinal mucosa were examined by HE staining and complement 3 was detected by immunohistochemical analysis. RESULTS: Compared with SO group, the vascular permeability, the activity of MPO and the content of MDA in I/R group were significantly increased, and the activity of SOD was decreased. HE staining demonstrated that I/R induced severe intestinal histological damages and the increased amount of complement 3 and its derivates were deposited in the necrosis area. Compared with I/R group, the vascular permeability, the activity of MPO and the content of MDA were decreased and the activity of SOD was significantly increased in CR1-SCR15-18 treatment group. CR1-SCR15-18 also significantly attenuated intestinal histological injury, and reduced the deposition of complement 3 and its derivates in the necrosis zone. CONCLUSION: sCR1-SCR15-18 protein exerts a protective effect against intestinal I/R injury in rats, possibly by inhibiting the activation of complement.     

Antigens from the midgut membranes of Ornithodoros erraticus induce lethal anti-tick immune responses in pigs and mice

Ornithodoros erraticus is an argasid tick that can transmit severe diseases such as human relapsing fever and African swine fever. In southern Europe O. erraticus lives in close association with swine on free-range pig farms. Application of acaricides for the eradication of O. erraticus from pig farms is inefficient. This is the reason why we tried to develop an anti-O. erraticus vaccine as alternative method of control. Accordingly, we were prompted to investigate the protective possibilities of a midgut membrane extract from the parasite (GME) that has not been studied hitherto. Administration of the GME with Freund's adjuvants (FAs) to pigs and mice induced a protective response able to kill 80% of the immature forms of the parasite in the first 72 h post-feeding and to reduce the fecundity of females by more than 50%. The action of the vaccine is the result of damage to the midgut wall of the argasid, and, in mice, it has been shown that this damage is mediated by activation of the complement system. In pigs, the administration of GME with alum, instead of with FAs, reduced the degree of protection. The protective antigens of the GME were expressed by all the developmental stages examined and are probably proteins from the luminal membrane of midgut epithelial cells. These antigens were seen to be more abundant in recently fed parasites than in fasting specimens, suggesting that their expression is induced after blood ingestion.     

复方中草药“强普素”对断乳仔猪血清补体4（C4）的影响

目的：研究复方中草药“强普素”对断乳仔猪血清C4的影响。方法：选择48头平均体重为（8.30±0.94）kg的（28±1）日龄杜×长×大断乳仔猪,按照随机区组设计分为4组,每组2个重复,每个重复6头仔猪（公母各半）。4个组分别为：（1）基础日粮（空白对照组）;（2）基础日粮+0.05%强普素（0.05%强普素组）;（3）基础日粮+0.1%强普素（0.1%强普素组）;（4）基础日粮+0.2%强普素（0.2%强普素组）。结果：添加0.1%和0.2%强普素的试验组仔猪血清C4水平显著高于空白对照组仔猪血清C4水平（P<0.05）,添加0.05%强普素的试验组仔猪血清C4水平与空白对照组仔猪血清C4水平差异不显著（P>0.05）。结论：强普素能提高断乳仔猪血清中C4的含量。     

Interaction of nucleophilic compounds with complement component C4

Drugs which induce systemic lupus erythematosus as a toxic side effect have been shown to inhibit the covalent binding of C4, which is an important event in immune complex clearance in normal individuals. Human C4 is encoded at two polymorphic loci, C4A and C4B within the Major Histocompatibility Complex and patients with idiopathic SLE are more likely to have a non-functional (null) C4A gene. The C4A and C4B gene products differ in reactivity with C4A being more reactive with nitrogen nucleophiles, including hydralazine and isoniazid (drugs which induce SLE), than with oxygen nucleophiles. We have established an assay system which allows the effect of nucleophiles on C4 in animal sera to be investigated. It has been found that in comparing reactivity of guinea-pig C4 with human C4A and human C4B that guinea-pig C4 is like human C4A and shows greater reactivity towards nitrogen nucleophiles than towards oxygen nucleophiles. This suggests that the guinea-pig should be a good animal model for drug-induced SLE.     

肌注PHA对犬血清中补体C3、C4的影响

为研究植物凝集素(PHA)对各犬血清中补体C3、C4浓度的影响,试验分成4个组:低剂量组(0.05 mg/kg)、中剂量组(0.10 mg/kg)、高剂量组(0.20 mg/kg)、对照组,对犬进行肌肉注射。在注射前和注射后的7 d、14 d对各组犬进行采血测定其血清中补体的含量,并进行统计分析。结果:中剂量组可显著提高机体的补体水平(P<0.05),并在高水平维持较长时间;低剂量组和高剂量组作用不显著(P>0.05)。     

Analysis of glomerular complement C5b-9 deposits and synthesis of NO,TNFα in the model of the rats with anti-thymocyte serum nephritis

AIM: To explore the localization and semi-quantification of the glomerular complement C5b-9 complexes and synthesis of some inflammatory mediators or cytokines such as nitric oxide (NO) and tumor necrosis factor α(TNFα) in the rats with anti-thymocyte serum nephritis(ATSN). METHODS: The animal model of rat ATSN was reproduced by a single intravenous injection of anti-thymocyte serum (ATS). Then, the deposits of glomerular C5b-9 complexes were localized and quantified by immunohistochemical staining and microscopic image scanning separately. And the glomerular mesangial cells (MC) surrounded by C5b-9 complexes were counted under microscope. In addition, the expression of glomerular MC inducible NO synthase(iNOS) mRNA and excretion of urinary NO metabolite (NO^-₂／NO^-₃) and TNF α in the rats with ATSN were detected. RESULTS: The MC in the rats with ATSN emerged necrosis followed by a rapid proliferation. In the early time of MC injury, the C5b-9 complexes were mainly seen in glomerular mesangium and MC surface. But with the progression of ATSN, the MC enclosed by C5b-9 appeared gradual decrease. Moreover, the expression of MC iNOS mRNA in early stage of ATSN obviously increased and the excretion of urinary NO^-₂／NO^-₃ and TNF α also significantly increased. However, the changes of parameters mentioned above in ATSN proliferative stage (after 7 days) alleviated gradually. CONCLUSION: The secondary lysis of MC has relation to the deposition of C5b-9 complexes and synthesis and release of NO and TNF α in rats with ATSN.     

Diagnosis of brucellosis by serology

Serological diagnosis of brucellosis began more than 100 years ago with a simple agglutination test. It was realized that this type of test was susceptible to false positive reactions resulting from, for instance, exposure to cross reacting microorganisms. It was also realized that this test format was inexpensive, simple and could be rapid, although results were subjectively scored. Therefore, a number of modifications were developed along with other types of tests. This served two purposes: one was to establish a rapid screening test with high sensitivity and perhaps less specificity and a confirmatory test, usually more complicated but also more specific, to be used on sera that reacted positively in screening tests. This led to another problem: if a panel of tests were performed and they did not all agree, which interpretation was correct? This problem was further compounded by the extensive use of a vaccine which gave rise to an antibody response similar to that resulting from field infection. This led to the development of an assay that could distinguish vaccinal antibody, starting with precipitin tests. These tests did not perform well, giving rise to the development of primary binding assays. These assays, including the competitive enzyme immunoassay and the fluorescence polarization assay are at the apex of current development, providing high sensitivity and specificity as well as speed and mobility in the case of the fluorescence polarization assay.     

A comparative study on urinary complement C5b-9 complex excretion in four nephritis models of rats

AIM: To explore the significance of measuring urinary complement C5b-9 complex in various types of immune complex (IC) nephritis models of rats. METHODS: The four models of rats, namely, passive Heymann nephritis (PHN), anti-thymocyte serum nephritis(ATSN), anti-glomerular basement membrane nephritis (AGBMN) and chronic serum disease nephritis (CSDN) were reproduced. Then, the contents of complement C5b-9 complex in plasma and urine of the rats were detected with sandwich ELISA. And the deposits of C5b-9 complex in glomeruli of the rats were examined by ABC immunohistochemistry staining. CONCLUSION: Urinary C5b-9 complex excretion could be taken as one of several sensitive immunologic parameters in diagnosing of PHN and in distinguishing PHN from other type of nephritis.