Haemagglutination and Haemagglutination-Inhibition Reactions of Extracts from Snails and Sponges

Abstract. The results of agglutination and agglutination-inhibition experiments with two spong extracts ( Axinella sp. and Cliona celata ) and three snail extracts ( Achatina granulata, Theba pisana and Helicella uirgata , banded form) are reported.
The extracts which show D-galactose specificity and are also inhibited by sugars related to D-galactose or containing it, have been subdivided into 1. a subgroup with no blood group specificity to which belong Axinella sp., Cliona celata and Ricinus communis extract and 2. another subgroup with B-like specificity to which belong extracts of Helicella virgata (banded form), Bandeiraea simplicifolia and Fomes fomentarius extracts.
Extracts of subgroup 1. but not of subgroup 2. are inhibited by Pn XIV poly-saccharide. In spite of anti-B-like specificity, extracts belonging to subgroup 2. are just as well or even better inhibited by NAcgal than by D-galactose.
The extracts of Achatina granulata and Theba pisana lack D-galactose specificity. Achatina granulata extract was inhibited by NAcgal, NAcglu and NANA; Theba pisana extract only by NANA.
Theba pisana extract agglutinated only neuraminidase-treated red cells, Helicella virgata (banded form) extract only enzyme-treated B cells.     

Haemagglutination and Haemagglutination-Inhibition Reactions of Extracts from Snails and Sponges

Abstract. Agglutination-inhibition, precipitation and precipitation-inhibition tests have been performed with 2 sponge and 4 snail extracts using as inhibitors a number of biological materials and some glycoproteins and glycolipids. Some human salivas and fluids from ovarian pseudomucinous cysts inhibited, others did not inhibit. Inhibiting power was unrelated to presence of ABH substances.
Some snail and seed extracts inhibited haemagglutination by Axinella sp. extract and some snail extracts haemagglutination by Cliona celata extract. Axinella sp. extract precipitated non-haemagglutinating Achatina granulata and Hedleyella falconeri snail extracts; this precipitation was inhibited by the same sugars which also inhibit haemagglutination by Axinella sp. extract.     

旋毛虫成虫三种抗原SDS-PAGE及Western-blot比较研究

目的研究大理猪源旋毛虫(Trichinella spiralis)成虫虫体可溶性抗原、表面抗原和排泄分泌抗原3种抗原的蛋白组分及其之间的差异,比较分析其免疫反应性。方法以十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和蛋白印迹对成虫3种抗原进行蛋白组分及其免疫反应性分析。结果十二烷基硫酸钠-聚丙烯酰胺凝胶电泳结果:虫体可溶性抗原显示29条蛋白带,分子量范围在112kDa~10kDa之间,其中主带13条;表面抗原显示4条主要蛋白带,分子量分别为50、48、40、34kDa;排泄分泌抗原显示17条蛋白带,分子量范围在120~14kDa之间,其中主带6条,分子量分别为120、64、43、40、35、33kDa。蛋白印迹结果:虫体可溶性抗原出现13条反应条带,其中分子量为81、40、37、33、30kDa的条带着色明显;表面抗原出现4条反应条带,其分子量分别为50、48、40、34kDa;排泄分泌抗原显示7条反应带,其中43、40、27、18kDa着色明显。结论旋毛虫成虫虫体可溶性抗原与排泄分泌抗原较表面抗原蛋白组分复杂,3者主带部分属低分子量区;免疫反应性的强度表面抗原和排泄分泌抗原高于虫体可溶性抗原,说明前两种抗原是旋毛虫成虫刺激机体产生免疫应答的主要靶抗原。     

湖北与云南两省钉螺酯酶同工酶酶谱的比较研究

采用聚丙烯酰胺凝胶垂直平板电泳，对湖北阳新、江陵两地的肋壳钉螺和云南大理，楚雄、丽江三地的光壳钉螺进行了酯酶同工酶的比较研究。五地钉螺共分离出１２条酶带，其中湖北两地的肋壳钉螺共分离出９条酶带，而云南三地的光壳钉螺共分离出１１条酶带。两省钉螺的酶谱间存在较大的差异，表现为７条酶带的性质不同和１条酶带的染色深浅不同。云南省三地的光壳钉螺的酶谱间仅略有不同，而湖北省两地的肋壳钉螺的酶谱则基本相同。因此认为，钉螺酯酶同工酶酶谱的特征能较好地反映钉螺属内的亲缘关系。该研究结果对今后研究钉螺的遗传距离，从而定量分析钉螺属内的亲缘关系，并为钉螺的分类提供了参考资料。     

Studies with Ferritin-Labelled Dolichos biflorus Lectin on the Numbers and Distribution of A Sites on A1 and A2 Erythrocytes, and on the Nature of its Specificity and Enhancement by Enzymes

S ummary . The conjugation of Dolichos biflorus extract with ferritin produced an electron dense anti-A reagent that was used to study the number and distribution of A sites on A₁ and A₂ red cells; there were five times as many ferritin-labelled lectin molecules on A₁ as on A₂ cells (about 800,000: 150,000). The arrangement of sites on both A₁ and A₂ cells was diffuse but not random, as pattern analysis of the ferritin molecules revealed a contagious distribution of the sensitized A sites. It is suggested that the anti-A₁ saline test activity shown by the Dolichos reagent is due to the inefficiency of the small lectin molecules to bring about agglutination of the A₂ cells. A₂ cells have fewer sites and consequently a larger mean distance between their A sites than have A₁ cells. Thus the probability of 'bridging' between the A sites of adjacent cells is reduced. Enzyme treatment increased lectin uptake and the A₂ and B cells both bound over 8 million molecules per cell. The A₂ cells gave agglutination reactions like untreated A₁ cells but the B cells were not agglutinated. In areas of agglutination between the cells lectin molecules probably occurred mainly as a monolayer; however, in the other areas they occurred in multiple layers and only the first layer was in contact with cell membrane sites.     

日本血吸虫门脉内童虫表膜抗原组分及其保护性免疫力研究

目的探讨日本血吸虫门脉内童虫表膜抗原(SjHmAg)的免疫特性,观察其抗日本血吸虫(Sj)的保护效果。方法用SDS-PAGE电泳技术分析SjHmAg蛋白组分,酶联免疫电转移印迹(EITB)分析感染兔血清(IRS)和正常兔血清(NRS)对SjHmAg的识别;用完整SjHmAg免疫昆明鼠3次,分别在0、2、4周进行,第6周每鼠经腹部感染40±1条Sj尾蚴,42天后剖杀,计数虫数及肝卵数。结果用SDS-PAGE电泳获得SjHmAg主带7条,IRS主要能识别SjHmAg23、33和63kDa等10个抗原组分;间接ELISA测其抗体滴度>1:6400,与对照组相比,SjHmAg免疫小鼠的减虫率为16.2％,减卵率为55.4％。结论用SDS-PAGE获得了不同分子量的SjHmAg蛋白,EITB鉴别出具有免疫活性的蛋白分子,且SjHmAg对Sj攻击感染及雌虫生植似有一定的抗性。     

黄果茄杀灭钉螺有效成分的分离提纯及其效果观察

目的分离提纯黄果茄果实灭螺的有效成分,观察灭螺效果,并确定其分子结构。方法将黄果茄果实去蒂去籽后干燥粉碎,用95%乙醇提取并浓缩,然后加乙酸乙酯处理,用薄层层析法(TLC)和柱层析法(CCL)对其进行分离,得到7种主要组份。采用“浸泡法”对其中4种含量较高的组分进行灭钉螺实验,确证Rf=0.58这一生物碱组分(A)为主要灭螺成分之一。选择最佳洗脱剂对灭螺效果最好组分A进行分离,得到A1及A2两组分。对A1,A2进行灭钉螺实验,比较其灭螺效果。电子轰击质谱测定有效成分分子量、核磁共振仪和红外光谱仪测定分子结构。结果黄果茄果实有效成分A(Rf=0.58)的灭螺效果较好,当投药量为2.50 mg/L,钉螺的死亡率为94.2%(28℃)。乙酸乙酯-氯仿-甲醇(11∶11∶35)系统对A的分离效果最好。灭螺对照实验发现:当A2投药量为0.20 mg/L,钉螺的死亡率为100.0%(28℃),其分子量为867,熔点为298～305℃。结论黄果茄有效成分提纯物A2为边缘茄碱(-αsolamarrgine),其对湖北钉螺有很好的杀灭效果。     

1993~2013年南京市钉螺及哨鼠血吸虫感染监测结果分析

目的 分析1993~2013年南京市21年来钉螺感染率与哨鼠血吸虫感染情况以及两者之间的相关性,为制定防治措施提供科学依据。 方法 1993~2013年每年春季采用系统抽样结合环境抽查法开展螺情调查,并在同一时间段采取小白鼠接触法对水体的血吸虫感染性进行现场检测。 结果 钉螺感染率、感染性钉螺面积总体呈逐年下降趋势,2009年至今未发现阳性螺点;哨鼠感染率1998年以前高达100%,2008年至今哨鼠感染率均为0。两者变化趋势一致,存在显著的正相关,相关系数rs为0.726,有统计学意义（P=0.000）。 结论 加大综合防治力度,控制感染性钉螺,依然是该市控制血吸虫病流行的有效防治措施。     

Comparative evaluation of freeze-dried and liquid antigens in the direct agglutination test for serodiagnosis of visceral leishmaniasis (ITMA-DAT/VL)

OBJECTIVE: The direct agglutination test (DAT) for visceral leishmaniasis (VL) with liquid (LQ) antigen is known to be only moderately reproducible because of inter-observer and batch-to-batch variability as well as its sensitivity to temperature and shaking during transport. We evaluated a DAT with freeze-dried (FD) antigen and compared it with the LQ antigen version. METHODS: Blood samples of clinical VL suspects and healthy endemic controls were collected in Sudan, Nepal and India. Both test versions were performed in duplicate in the respective countries and in the reference laboratory. Interbatch variability and stability tests were conducted and agreement was examined within and between centres on a dichotomic scale by Cohen's kappa as well as on a continuous scale through Bland-Altman plots. RESULTS: The FD antigen remains fully active even after storage at 45 degrees C for 24 months. Using a cut-off titre of 1:6400, the agreement between the FD and the LQ formats was excellent. CONCLUSION: The major advantages of FD antigen are its better stability at higher temperatures and its longer shelf life, which make it much more suitable than the LQ version for use in the field.     

Sero-epidemiological survey of hepatitis virus infection in Taiwan: A study in 6863 subjects from the general population

Abstract A sero-epidemiological study was made of 6863 apparently healthy randomly selected volunteers from various parts of Taiwan. It revealed an 18% incidence of hepatitis B surface antigen (HBsAg) carriers amongst the general population, the prevalence being significantly higher in males than females (21.7% vs 14.1%). The Takasago natives showed the highest HBsAg carrier rate (27.0%). Those who are primarily derived from Ming Nan had a significantly higher prevalence (19.0%) than those from Hakka (14.7%) and recent immigrants from Mainland China (14.6%). About one-third of female carriers of child-bearing age in southern Taiwan were positive for e antigen (HBeAg) in a limited study; they may be an important source of HB virus (HBV) infection. The positivity rate for HBeAg was significantly higher in subjects below 40 years of age (36.7%) than those above 40 years (7.5%). There was a significant association between HBsAg positivity and elevated serum transaminase levels, especially in males; 20.3% of carriers had elevated transaminase in comparison with 11.1% of female carriers. Among HBsAg seronegative individuals, the frequency of abnormal transaminase increased with age with an overall abnormality rate of 10.6% in males and 5.0% in females, suggesting that factors other than HBV may also contribute to liver disease in Taiwan. Altogether, 9.3% had elevated transaminase. Out of 82 randomly selected HBsAg carriers who were evenly distributed in geography, serum antidelta antibody was positive in only five men (6.1%), of whom four had a mild elevation of transaminase levels (< 100 KU). The overall positivity rate for anti-hepatitis A (HA) antibody was 93.0% in southern Taiwan, indicating poor sanitation. These findings indicate that HB and HA virus infections are highly endemic in Taiwan and that delta infection is infrequent despite a high prevalence of HB virus infection.     

Prothrombin antigen levels in symptomatic and asymptomatic carriers of the 20210A prothrombin variant

A recently discovered variant in the prothrombin gene (20210A) has been found in approximately 5–10% of patients with venous thromboembolism. It has been shown that patients with this variant present with high levels of prothrombin in plasma and this is maintained to be the most likely mechanism by which the risk of thrombosis is increased. We have evaluated prothrombin antigen levels in 50 carriers of the 20210A allele and compared with non-carriers. 327 subjects were subdivided according to deficiency status and previous thrombosis. 30 symptomatic and 20 asymptomatic carriers had increased mean prothrombin levels as compared to symptomatic ( n  = 178) or asymptomatic ( n  = 99) non-carriers. The percentage of subjects with prothrombin levels above cut-off values of 1.15 u/ml or 1.30 u/ml was significantly higher in carriers of the prothrombin variant as compared to non-carriers, regardless of a previous thrombosis. However, among non-carriers the percentage of those with prothrombin levels above cut-off values was significantly higher in the group of symptomatic as compared to asymptomatic individuals. In conclusion, increased prothrombin antigen levels, as detected by a specific ELISA, were found among 50 symptomatic and asymptomatic carriers of the 20210A prothrombin variant as well as among a large group of symptomatic non-carriers. The data are in agreement with those found by using functional tests for the determination of prothrombin levels in these patients.     

Comparison of the Clinical Features of Hepatitis A between HBsAg-Positive and HBsAg-Negative Patients

Background/Aims

The notion that acute hepatitis A superimposed on chronic hepatitis B infection leads to a worse outcome than acute hepatitis A alone remains controversial. The aim of this study was to determine the influence of the presence of hepatitis B surface antigen (HBsAg) on the severity of acute hepatitis A.

Methods

We retrospectively analyzed 449 patients hospitalized for acute hepatitis A from January 2000 to February 2010 and compared clinical outcomes based on the presence of HBsAg.

Results

Of the 449 patients, 30 patients were in the HBsAg-positive group and 419 in the HBsAg-negative group. The HBsAg-positive group was older than the HBsAg-negative group (36.1±8.3 vs 31.8±8.5 years, p=0.004); however, other baseline characteristics were similar between the 2 groups. Mean peak values of prothrombin time, serum total bilirubin, and serum creatinine at admission were significantly higher in the HBsAg-positive group. When comparing clinical outcomes between the 2 groups, gastrointestinal bleeding, acute renal failure, and acute liver failure were more frequently observed in the HBsAg-positive group. In particular, the incidence of acute liver failure was approximately 9-fold higher in the HBsAg-positive group than in the HBsAg-negative group (23.3% vs 3.3%; odds ratio [OR], 8.80; p<0.001). Multivariate analysis showed that HBsAg (OR, 7.43; 95% confidence interval [CI], 2.56 to 21.57) and age (OR, 1.07; 95% CI, 1.02 to 1.13) were independent risk factors for the occurrence of acute liver failure.

Conclusions

In patients with chronic hepatitis B infection, acute hepatitis A is associated with more severe clinical outcomes, including acute liver failure, compared with patients with acute hepatitis A alone.     

Analysis of density and epitopes of D antigen on the surface of erythrocytes from DEL phenotypic individuals carrying the RHD1227A allele

Background

The characteristics of the D antigen are important as they influence the immunogenicity of D variant cells. Several studies on antigenic sites have been reported in normal D positive, weak D and partial D cases, including a comprehensive analysis of DEL types in Caucasians. The aim of this study was to assess D antigen density and epitopes on the erythrocyte surface of Asian type DEL phenotypic individuals carrying the RHD1227A allele in the Chinese population.

Materials and methods

A total of 154 DEL phenotypic individuals carrying the RHD1227A allele were identified through adsorption and elution tests and polymerase chain reaction analysis with sequence-specific primers in the Chinese population. D antigen density on the erythrocyte surface of these individuals was detected using a flow cytometric method. An erythrocyte sample with known D antigen density was used as a standard. Blood samples from D-negative and D-positive individuals were used as controls. In addition, D antigen epitopes on the erythrocyte surface of DEL individuals carrying the RHD1227A allele were investigated with 18 monoclonal anti-D antibodies specific for different D antigen epitopes.

Results

The means of the median fluorescence intensity of D antigen on the erythrocyte membrane surface of D-negative, D-positive and DEL individuals were 2.14±0.25, 193.61±11.43 and 2.45±0.82, respectively. The DEL samples were estimated to have approximately 22 D antigens per cell. The samples from all 154 DEL individuals reacted positively with 18 monoclonal anti-D antibodies specific for different D antigen epitopes.

Discussion

In this study, D antigen density on the erythrocyte surface of DEL individuals carrying the RHD1227A allele was extremely low, there being only very few antigenic molecules per cell, but the D antigen epitopes were grossly complete.     

日本血吸虫童虫细胞与非细胞型免疫原在小鼠免疫部位滞留的动态观察

目的观察日本血吸虫(Schistosoma japonicum,Sj)童虫细胞型免疫原和童虫细胞碎片免疫原免疫小鼠后在免疫部位滞留的动态变化。方法昆明鼠实验组A和B经大腿肌肉分别注射107日本血吸虫原代童虫细胞(pJCs)和相当剂量的童虫细胞碎片(JCFs),对照组C注射PBS。分别在注射后1、3、6、9和12 d,各组随机处理4或2只小鼠,从注射部位定量获取肌肉组织,用Western blot法和免疫组化法观察、比较两种类型抗原滞留的动态变化;用组织病理学方法观察发生的炎症反应。结果Western blot显示,A组在注射后各时间点均检测到分子质量单位为75 ku的特异性抗原条带,B组在注射后第6 d该条带即消失,C组未检出该条带。免疫组化检测结果与Western blot相符。组织病理学检查结果显示,A组在免疫后第12 d仍可见肌细胞间隙内大量炎性细胞浸润,B组在免疫后第12 d炎性细胞基本消失,C组未见炎性反应。结论血吸虫童虫细胞型免疫原较童虫细胞碎片免疫原在免疫部位可滞留更长时间,引起更持久的炎性反应,这可能是其诱生高保护性抗血吸虫病免疫的重要机制之一。     

2004~2015年湖南汉寿县垸内钉螺控制措施及效果分析

【摘要】 目的  探索湖沼型垸内亚型疫区的垸内钉螺控制对策。方法 收集汉寿县血吸虫病流行及防治档案资料,对2004~2015年汉寿县垸内钉螺指标变化、垸内钉螺控制的主要措施及效果进行评价。结果 汉寿县垸内钉螺面积由2004年的99.64hm2减少到2015年的0.87hm2,下降幅度达99.1%;活螺平均密度由2004年的0.15只/0.11m2下降至2015年的0.03只/0.11m2;感染性钉螺处数与面积显著下降,2006~2015年未查获感染性钉螺。结论  在药物灭螺的基础上,针对垸内钉螺采取水改旱、养殖灭螺、沟渠硬化、国土平整、涵闸改造、小环境治理等措施,对控制并消灭垸内钉螺有显著性作用。为实现消除血吸虫病目标,各级政府应继续加大血吸虫病综合治理项目经费的投入。     

神经肽Y及其受体与某些老年病及作用机制的研究

高血压、冠心病、肺心病、支气管哮喘、慢性阻塞性肺病、糖尿病、老年性痴呆、帕金森氏病、老年抑郁症与焦虑症、骨质疏松、肿瘤等均是老年人较为常见的疾病,它们直接或间接地威胁甚至危害着老年人的健康和生命。然而,大量基础医学和临床研究发现,神经肽Y及其受体在机体的分布、含量、合成、分泌及表达水平和调控功能等,不仅有可能介导或影响着这些老年性疾病的发生、发展、加重或预后,而且还有可能与治疗这些疾病的生物学效应、作用靶点、用药途径及治疗对策等密切相关。因此,本文就神经肽Y及其受体与这些老年病之间的关系及其作用机制方面的研究进展作一综述,为今后探索以神经肽Y及其受体为靶点的有效治疗方式奠定基础,也为老年医学的科学研究和临床防治提供一些理论和实验依据。     

卡介苗(BCG)D2株分泌型抗原85A基因的克隆与序列分析(英文)

目的　分离BCGD2株分泌型抗原 85A基因并测定DNA序列 ,为研制抗结核病新型疫苗奠定基础。方法　PCR法从BCGD2株基因组DNA中分离Ag85A基因 ,PCR产物连接入 pUCm -T载体 ,重组克隆用单菌落PCR法、BamHⅠ和SacⅠ双酶切以及DNA测序进行鉴定。结果　分泌型Ag85A基因经过分离和测序后发现 ,它有 10 4 1bp组成 ,与LukDeWit等人测定的来自于BCG1173P2株的同一基因的DNA序列是一致的 ,表明Ag85A基因在分枝杆菌中是高度保守的。 结论　BCGD2株分泌型抗原 85A基因的成功克隆与序列分析将会促进对Ag85A基因深入的研究 ,以及为研制抗结核病候选疫苗奠定基础