成人肾脏ABO组织血型1-4型糖链H物质的分布

目的观察1型H、2型H及3/4型H糖链在成人肾组织中的分布及其与分泌状态的关系。方法 应用抗ABO抗体及3种糖链特异的单克隆抗H抗体的免疫组织化学方法,检查分泌型与非分泌型个体肾组织中相应抗原物质的分布。结果 在分泌型和非分泌型人的肾远曲小管均表达2型H和3/4型H物质,1型H和3/4型H物质只在分泌型人肾集合管表达,在非分泌型人中不表达。另外集合管的2型H物质的表达与分泌状态无关。结论 人肾组织有ABH物质的表达,不同肾组织细胞表达的H物质结构差异与AB0型分泌状态有关。     

非分泌型的基因型与血型物质的分泌研究

阐明非分泌型的基因型与血型物质分泌量和Lewis表型的关系。应用时间决定性荧光免疫测定法（TR－FIA），检测传统的A型Lewis阳性非分泌型个体唾液中H、A及Lewis抗原含量，并以序列特异性PCR，确定其基因型。非分泌型个体唾液中检出了高含量的Lea抗原，其中8例不同程度的检出了少量H、A和Leb抗原，其FUTZ位点为se2／se2纯合型，属Le（a＋b＋）型；1例基因型为se3／se5杂合型，未能检出H、A及Leb抗原，属Le（a＋b－）型。se2是弱分泌基因，se3及se5是非分泌基因。     

Determination of ABH secretor status by an electronic quantitation method

Blood Group A and B substances in secretor (Se) and nonsecretor (se) salivas were tested by means of an electronic data processing-hemagglutinin-inhibition test (EDP-HAIT) with immunoglobulin M (IgM) isohemagglutinins. Besides a difference in quantity, the blood group substances in Se saliva showed high binding efficiencies compared with those in se saliva. EDP-HAIT with IgG isohemagglutinins proved no difference in the binding efficiencies of Se and se salivas. The determination of secretor status by EDP-HAIT with IgM isohemagglutinin was accurate because the conclusion was obtained based on two different quantitative results. Secretor status of some salivas in gargled water could be determined by comparing the binding efficiencies.     

A, B and H group specific substances in human vitreous humor

The presence of A, B and H group specific substances in vitreous humor taken from 105 human corpses was determined. Good agreement was obtained between these group substances and the ABO blood group. The relationship with the secretor type is discussed.     

Enzyme-linked immunosorbent assay for A and B water soluble blood group substances

The conditions affecting an enzyme-linked immunosorbent assay for salivary blood group substances were investigated. It was found that A, B, and O secretor saliva samples would each bind both anti-A and anti-B typing reagents. The conditions that affected the assay response were optimized for maximum sensitivity and to give the highest resolution possible between the result for an antiserum binding to homologous antigen and the response for heterologous antigen-antibody combinations. Monoclonal antibodies eliminated the heterologous binding indicating that this binding was due to a lack of specificity of the routine typing reagents. A sensitive assay using the monoclonal antibodies to distinguish between samples of A and B secretor saliva is described.     

唾液血型抗原分泌时限变化及温度对抗原活性的影响

探讨人唾液中ABH血型抗原不同时限的分泌量,以及保存温度对血型抗原活性的影响。应用时间决定性荧光免疫测定法（TR．FIA）对O型分泌型10例和非分泌型5例人在不同条件下唾液中H抗原量进行检测。唾液血型抗原的分泌量随时间而波动,进餐后降低明显,但不干扰分泌型的判定。37℃保存48h抗原活性完全丧失,6℃保存1周抗原活性几乎没有变化。结果表明,唾液分泌时段不影响分泌型判定,将唾液制成斑痕可长期保存样品。     

The use of anti-A and anti-B monoclonal antibodies in forensic medicine

Anti-A and anti-B monoclonal antibodies (MCA) may be used in the analysis of liquid blood, blood and saliva traces in order to detect ABO blood group antigens A and B using common methods of evidence investigation. Use of MCA and isohaemagglutinins anti-A and anti-B in absorbtion-elution reaction during the analysis of minute saliva traces enhances the possibility of establishing the origin of the saliva at the expense of nonsecretor.     

H-抗原缺乏分泌型个体FUT1及FUT2等位基因的分子基础

一例H-缺乏分泌型个体被发现。其血清学表现为：红细胞上无A、B、H抗原；唾液中有B、H抗原分泌；血清中检出了抗A抗体。推测其为类孟买OHmB型。在该个体FUT1等位基因编码区上发现两处单碱基突变（T460C、G1042A）。这两个点变异，将导致两个氨基酸的置换（Y154H、E348K）。同时也破坏了限制性内切酶RsaⅠ和AvaⅠ的作用部位。用PCR－RFLP法可检出此两种变异。用PCR－RFLP法证实，该个体为T460C、G1042A变异的纯合子。在136例随机个体中未能查出上述变异。将该FUT1基因转染COS－7细胞未能检出α2－FUT活性及证实H抗原的表达。该个体的FUT2基因与野生型一致。     

人体组织ABO血型检测方法的研究

探讨人体组织ABO血型检测方法。对已知ABO血型尸体的不同组织,用红细胞粘连试验、吸收—抑制试验和吸收－解离试验进行ABO血型测定。12例尸体的16种组织中均检出与尸体血痕相同的ABH物质。对不同温度保存的组织块进行ABH物质检测的结果显示,4℃保存的组织ABH物质的检出时间长于室温,空腔脏器的检出时间短于实质脏器。三种方法中,红细胞粘连试验简单易行,适用于基层单位,吸收－抑制试验用于组织块的测定时优于吸收－解高试验。     

The rapid determination of the ABO group from body fluids (or stains) by dot enzyme-linked immunosorbent assay (dot-ELISA) using enzyme-labeled monoclonal antibodies

Using ABH enzyme-labeled monoclonal antibodies, the authors could rapidly detect the ABO group from body fluids and body fluid stains by the dot enzyme-linked immunosorbent assay (dot-ELISA). In this test, the antigen was immobilized on nitrocellulose paper; the entire piece of paper was coated with an appropriate dilution of enzyme-labeled McAb directly against the antigen of interest; and, finally, 3,3'-diaminobenzidine (DAB) substrate solution was added. The site of a positive reaction is clearly visible as a brown spot. We analyzed 521 samples and got satisfactory results. We also analyzed 99 practical case samples by this method and achieved the same results as those obtained by other researchers using other methods. This method is accurate, simple, direct, rapid, and sensitive; it also produces easily observed results, requires no equipment, and can be completed in 30 min. The test proved to be clearly more sensitive for the detection of the ABO blood group in secretor saliva than the conventional hemagglutination inhibition test. Also saliva diluted 10(-4) to 10(-5) and the ABO group of nonsecretor saliva and urine could be easily detected by this method.     

ABH and Lewis antigens in the tracheal glands. I. Lewis-positive individuals

Antigens A, B, H, Lea, and Leb were demonstrated in the tracheal glands of 15 Lewis-positive secretors and 15 nonsecretors by the indirect immunoperoxidase technique. The detection of group-specific ABH antigens in mucous epithelium and intraductal secretory fluid was dependent on the secretor character. Whereas determination of secretor character was sometimes unreliable with anti-A and anti-B, the findings obtained by additional labeling with UEA1 were consistently correct. The secretors showed minimal gland labeling with anti-Lea and intensive labeling with anti-Leb; the nonsecretors, intensive Lea labeling and weaker or absent Leb labeling. Consequently, the determination of secretor character by ABH labeling could be verified by the behavior of the Lewis antigens. Since both morphologic structures and epithelial antigens are highly resistant to putrefaction, ABO and secretor character can also be diagnosed in badly decomposed tracheal wall specimens.     

Gm(1,2,4,10,21) and Km(1) factors in vitreous humor

The presence of Gm(1,2,4,10,21) and Km(1) factors in vitreous humor taken from human corpses was investigated. The results revealed a good agreement between the factors detected in this biological material and in blood. Their presence in vitreous humor is independent of the secretor type.     

Forensic medical diagnostics of chronic alcoholic intoxication based on histological changes in the soft tissues of oral cavity and salivary glands

Histological studies of oral cavity mucosa and salivary glands in subjects with chronic alcoholic intoxication revealed changes at the surface of the tongue and in the glandular tissues. Specific features of chronic alcoholic intoxication include acinar and ductal hyperplasia, reduction of the adipose tissue mass in salivary gland stroma, predominance of T-lymphocytes in hard palate minor salivary glands and B-lymphocytes in the stroma of labial minor salivary gland, the absence of plasma cells in the stroma of hard palate minor salivary glands and labial mucosa. Leukoplakia, dysplasia, and hyperplasia of the basal epithelial layer of oral cavity mucosa are considered to be the signs of long-term (over 12 months) alcohol consumption.     

Detection of hereditary enzyme characteristics in vaginal secretion

Upon investigation of semen- and blood-free vaginal swabs using starch gel electrophoresis the Phosphoglucomutase type was clearly identified in about 40%. Using cellulose acetate membrane electrophoresis PGM could not be demonstrated. In all cases the results correspond with those obtained in blood. No relation was found between secretor type (determined in saliva) and PGM typing. In vaginal material the following could not be determined: Adenylatkinase (AK) using starch gel electrophoresis, Esterase D (EsD) using cellulose acetate membrane electrophoresis, and Glyoxalase I (GLO) using agarose gel thin-layer electrophoresis.     

Establishment of a group classification of isolated cells using anti-A and anti-B monoclonal antibodies

Anti-A and anti-B monoclonal antibodies may be used for determination of ABO system A and B antigens in mixed agglutination reaction. The authors suggested a variant of this reaction which ensures reliability of antigen detection in cells of people of secretor state. Investigation of mixed traces (liquid salivary part and buccal epithelium cells) showed that three-fold washing of cells couldn't provide for reliable identification of their blood group.     

The detection of gamma globulin and Inv-factors in semen and saliva and of haptoglobins in semen (author's transl)]

Gm- 1-, 2-, and Inv 1-factors can be demonstrated in semen and saliva. For the examination of traces it is necessary to verify the suitable dilutions for the different charges of antisera and to test the eluates of the samples if they have a sufficient concentration. We observed some incorrect negative results in seminal and saliva stains which were apparently caused by insufficient material. The demonstration was independent of the secretor type. Haptoglobin could not be determined in semen and saliva.     

Cytomegalic inclusion disease of the salivary glands in sudden infant death syndrome

The parotid gland and/or the submandibular gland in SIDS cases (180 from Berlin und 75 cases from Hamburg) were examined by means of HE staining, immunohistochemical analysis, in situ hybridization and electron microscopy. The SIDS cases were taken from the last 10 years; the age of the children ranged from 2 weeks to 1 year. Typical cytomegaly inclusion bodies were recognized in 10% (18 cases from Berlin; more girls than boys) and 7% (6 cases from Hamburg; more boys than girls). Our files indicate that the frequency of CMV infection was not age-dependent within the first 12 months of life. Using immunohistochemical analysis and in situ hybridization, virus substances were detected in cytomegal cells as well as in morphologically uninfected cells. The literature on the clinical and epidemiological aspects of cytomegaly indicates that a localized CMV infection of the salivary glands does not sufficiently explain the sudden death of these infants; however, it should be emphasized that cytomegaly can influence the immunological status of the organism.     

尸体甲状腺球蛋白降解及其与死亡时间的关系

目的探讨尸体甲状腺球蛋白降解程度与死亡时间(PM I)的关系。方法死后21d内尸体的甲状腺组织,经免疫组化(EPOS)法染色,观察甲状腺球蛋白的染色反应;对63例阳性染色的甲状腺组织(PM I≤5d)进行计算机图像分析,并对所得图像参数的数据进行统计学分析。结果PM I≤5d的甲状腺组织,甲状腺腺泡上皮细胞胞浆及甲状腺胶质中均显示了不同程度的免疫反应阳性;PM I超过12d,免疫反应阴性;经图像分析及统计学处理,其积分光密度、分布密度和目标面积与PM I之间的确定系数(R2)分别达0.9794,0.9732及0.9884。结论尸体甲状腺组织的甲状腺球蛋白降解程度随PM I不同而变化,二者呈现较强的相关性。     

Detection of ABH blood group antigens in the saliva of Koreans and their stability according to storage of saliva samples

The purpose of the present study was to identify salivary molecules carrying the ABH blood group antigens in Koreans and to investigate the changes in these antigens according to processing and storage of saliva samples. Secretor or non-secretor phenotypes and salivary components carrying the ABH antigens were identified in 90 subjects, 30 subjects in each ABO blood group, by SDS-PAGE and immunoblotting. Saliva samples were then obtained from 12 secretors-two males and two females in each ABO blood group and aliquots of both fresh saliva samples and their supernatants after centrifugation were stored at room temperature, 4, -20 and -70 degrees C. The same experiments were performed after 1, 3 and 6 months to investigate changes in the blood group antigens. In all 68 secretors, high-molecular-weight salivary mucin (MG1) was found to be the primary carrier of the ABH antigens. A salivary component of approximately 80 kDa also carried H antigen in seven saliva samples of 22 blood type O secretors. The blood group antigens were better detected in centrifuged samples. In saliva samples preserved at room temperature and 4 degrees C, the blood group antigens were either not detected or detected as degraded molecules. No change was found in the blood group antigens in saliva samples preserved at -20 and -70 degrees C for 6 months.     

Bloodgroup simulating activity in aerobic gram-negative oral bacteria cultured from fresh corpses.

The gram-negative aerobic oral bacterial flora of 100 consecutive corpses was isolated. After the identification and culturing of the isolated organisms, blood grouping was performed by the haemagglutination inhibition technique on dried culture smears, the dried culture medium and a dried ethanol extract of the bacteria. Forty-seven of the samples showed a gram-negative aerobic bacterial growth, giving 58 microorganisms of 14 different species. Positive blood grouping results were found in two of them (Escherichia coli and Serratia marcescens), both of type B. It is concluded that occasional mistyping of blood groups on saliva and oral material may be caused by the oral gram-negative aerobic flora, especially if the specimens are contaminated or putrefying.