Identification and Characterization of Peptide Mimics of Blood Group A Antigen

In order to investigate peptide mimics of carbohydrate blood group A antigen, a phage display 12-mer peptide library was screened with a monoclonal antibody against blood group A antigen, NaM87-1F6. The antibody-binding properties of the selected phage peptides were evaluated by phage ELISA and phage capture assay. The peptides were co-expressed as glutathione S-transferase （GST） fusion proteins. RBC agglutination inhibition assay was performed to assess the natural blood group A antigen-mimicking ability of the fusion proteins. The results showed that seven phage clones selected bound to NaM87-1F6 specifically, among which, 6 clones bore the same peptide sequence, EYWYCGMNRTGC and another harbored a different one QIWYERTLPFTF. The two peptides were successfully expressed at the N terminal of GST protein. Both of the fusion proteins inhibited the RBC agglutination mediated by anti-A serum in a concentration-dependent manner. These results suggested that the fusion proteins based on the selected peptides could mimic the blood group A antigen and might be used as anti-A antibody-adsorbing materials when immunoabsorption was applied in ABO incompatible transplantation.

Blood group type antigens in pancreatic intraductal papillary mucinous neoplasms

BACKGROUND: There are few data on blood group(BG) types and types of pancreatic cancers. The aims of this study were to study BG types and BG-antigens in pancreatic intraductal papillary mucinous neoplasms(IPMNs). METHODS: BG type and tumor BG-antigen(glycoprotein) expression(studied by immunohistochemistry on tissue microarrays) were analyzed with regard to characteristics of 101 surgically resected pancreatic IPMNs. RESULTS: Non-O BG type predicted invasive carcinoma independently from high serum CA19-9 and male gender. BG type A was observed more frequently in women than in men. Chronic pancreatitis was more frequently seen in patients with BG type B or AB. Aberrant tumor expression(with regard to BG type) of loss of A antigen expression type occurred in 15.0% of IPMNs and of loss of B antigen expression type in 62.5% of IPMNs. Intraneoplasm BG-antigen expression was not related to dysplasia grade or invasion. CONCLUSION: The results of the study suggest that in pancreatic IPMN, non-O BG type predicted invasive carcinoma, whereas for intratumor BG-antigen expression no specific patterns were detected with regard to the progression of glandular epithelial dysplasia or invasion.

Relationship of A,B,H blood group antigens with pathomorphological grading and prognosis of transitional-cell carcinoma of the urinary bladder

After being labelled with monoclonal antibodies against A,B, H blood group antigens,100 specimens of transitional-cell carcinoma of the urinary bladder were studied with ABC immunohistochemical technique and the cases were followed up.It was found that the overall positive rate of A,B,H antigens was 63%. The mortality rate and recurrence were significantly lower in the positive group than in the negative group(P<0.01)and 5-years survival rate was higher in the positive than in the negative(P<0.01).The findings suggest that the expression of blood group antigens is more effective for the prognosis of transitional-cell bladder carcinoma than the pathomorphological grading.

Effect of sodium nitroprusside on the microrheological properties of red blood cells in different media

Red blood cell(RBC)aggregation as well as their deformation significantly affects blood microrheology.These processes depend on various factors,one of which is concentration of the nitric oxide,one of the main signaling molecule in the bloodstream.The purpose of this study was to investigate the effect of nitric oxide on the microrheological properties of red blood cells(RBCs)in RBC samples of various media after the addition of nitric oxide donor sodium nitroprusside in vitro.Microrheological properties were measured using laser aggregometer and ektacytometer based on diffuse light scattering and diffraction of laser light on a suspension of RBCs,respectively.The study found that heparin-stabilized blood showed increased RBC aggregation and deformation with sodium nitroprusside concentrations of 100,and 200M,while EDTA-stabilized blood showed slightly decreased aggregation and unchanged deformation.With washed RBCs in dextran solution,the addition of sodium nitroprusside(in the concentrations of 100,and 200M)resulted in decreased aggregation and increased deformation.These-ndings aid in our understanding of nitric oxide's effect on RBC microrheological properties.

Significance of Blood Type Y Antigen Expression in Breast Adenosis and Cancer

The expression of blood type Yantigen in the epithelial cells from 46cases of breast adenosis,5 cases of malig-nant degeneration of adenosis,52 casesof primary breast cancer,and 5 cases ofnormal mammary tissue was a-ssessed with ABC immunoperoxidasehistochemical assay and Y hepten specificnomoclonal antibody.

Clinical significance of the detection of Rh blood group antigens and irregular antibodies in pregnant women with a second pregnancy

Objective:To investigate the phenotype distribution of five antigens of Rh blood group system and the specificity of Rh blood group irregular antibodies in pregnant women with second child.To analyze the relationship between Rh blood group antibody and hemolytic disease of the newborn(HDN)in second-child pregnant women,and to provide laboratory basis for the diagnosis and treatment of hemolytic disease of the newborn(Rh-HDN).Methods:500 pregnant women with second child were collected as the study group and 500 pregnant women with first pregnancy as the control group(all pregnant women underwent obstetric examination in the integrated obsteric clinic of our hospital from January 2020 to January 2021).To detectethe Rh blood group antigens(D,C,c,E,e)of the two groups of samples,screene the irregular antibodies,identify the specificity of irregular antibodies,determine the titer and record the hemolytic disease of the newborn of pregnant women with positive Rh blood group antibodies.Results:There were 11 Rh phenotypes in the pregnant women with second child in the study group:CCDee(152cases,30.4%),CcDEe(136cases,27.2%)CcDee(84cases,16.8%),ccDEE(30cases,6%),ccDee(31cases,6.2%),CCDEe(14cases,2.8%),ccDEe(9cases,1.8%),cc dee(18cases,3.6%),CCDEE(2cases,0.4%),CcdEe(12cases,2.4%),Ccdee(6cases,1.2%),CCd ee(6cases,1.2%).A total of 42 cases(8.4%)in the pregnant women with second child were negative for RhD.There were 10 Rh phenotypes in the pregnant women with first pregnancy in the control group:CCDee(144cases,28.8%),CcDEe(138cases,27.6%),CcDee(90cases,18%),ccDEE(42cases,8.4%),ccDee(28cases,5.6%),CCDEe(10cases,2%),ccDEe(8cases,1.6%),cc dee(19cases,3.8%),CCDEE(1cases,0.2%),CcdEe(11cases,2.2%),Ccdee(9cases,1.8%).A total of 39 cases(7.8%)in the pregnant women with first pregnancy were negative for RhD.In the pregnant women with second child in the study group,the positive rate of irregular antibody screening was 4.0%(20/500),and the specificity of Rh blood group antibodies was found as follows:anti-E 1.8%(9/500),anti-D 1.4%(7/500),anti-C 0.4%(2/500)and anti-Ec 0.4%(2/500).The positive rate of irregular antibody screening in the pregnant women with first pregnancy in the control group was 0,and the difference between the two groups was statistically significant(P<0.05).Rh-HDN was found in 10 newborns(2%)of the 20 women with positive irregular antibodies in the pregnant women with second child,and the antibody titer during pregnancy was more than 32.No Rh-HDN occurred in newborns in the pregnant women with first pregnancy,and the difference between the two groups was statistically significant(P<0.05).Conclusion:Pregnancy stimulation can increase the probability of irregular antibodies in pregnant women,and irregular antibodies in Rh blood group can easily cause Rh-HDN,so attention should be paid to routine detection of five antigens of Rh blood group and irregular antibody screening during prenatal examination.It is helpful for the early detection of Rh-blood irregular antibodies and the assessment of fetal or neonatal risk of Rh-HDN.

An Experimental Study on RBC Count and Serum Potassium Concentration Changes During Compression Transfusion of WBC-removal Whole Blood

Objective： To observe changes in RBC count, changes, RBC morphology, and serum potassium during compressed transfusion of WBC-removal whole blood. Methods： Prepared human WBC-removal whole blood and connected transfusion apparatus with different sizes of intravenous detaining needles （18G, 20G, 22G and 24G）. Observed RBC count and serum potassium concentration under different pressures （100 mmHg, 200 mmHg, and 300 mmHg） as blood flowed out of the pinhead end of the intravenous detaining needle. Samples obtained with the 20G needle were smeared on glass slides, and RBC morphologic changes were observed under an oil immersion lens. Results： RBC count and serum potassium changed slightly under different pressures with different sizes of intravenous detaining needles as blood flowed through the transfusion apparatus. In addition, the observation of blood samples under a common light microscope revealed that coarse- prick, oblong, and spindle cell counts in the visual fields increased gradually as the pressure increased. Additionally, a portion of cells had undergone splintering. Conclusions： While applying 18G, 20G, 22G and 24G intravenous detaining needles for blood transfusion under less than 300 mmHg of pressure, no significant RBC count change was found in blood samples in the short term. However, there were significant RBC morphologic changes. The results could offer more basis to ensure the clinical safetv of oatients undereoing blood transfilsion.

Tumor-Specific Histo-Blood Group Antigens: Apropos of Two Cases

Cancer cells with immunogenic properties having altered protein glycosylation, modified blood group substances have been widely studied. Due to the genetic instability occurring during carcinogenesis the glycosyltransferases may suffer from posttranslation sequence modification. The author describes 2 autopsy cases, where in the background of the unusual metastatic tumor presentation, incompatible blood group antigenic determinants have been demonstrated using blood group specific lectins and monoclonal antibodies (mAb). In the first case, reported here, a 10-year-old girl developed an acute myeloid leukemia and died in a septic endotoxin shock after successful cytostatic treatment of a juvenile signet ring cell cancer of her colon. At autopsy there were no signs of tumor except bilateral apple-sized mucinous ovarian (Krukenberg) metastases. While she had erythrocyte phenotype of blood group A, the signet ring adenocarcinoma cells expressed blood group B incompatible antigenic determinants with lectin/mAb. In the second case, the autopsy of a 78-year-old female resulted in no macroscopic tumor sign except a moderately enlarged, ham hard spleen. Light microscopy revealed adenocarcinomatous infiltration in the splenic sinusoids. The patient had blood group O, while the metastatic cells in the spleen reacted with Breast Carcinoma Antigen (BioGenex) and incompatible anti-B Banderiaeasimplicifolia agglutinin I and anti-B mAb. It proved to be a case of an occult, completely regressed breast cancer. Based on these observations the expression of tumor specific incompatible blood group antigens might occur from time to time, mostly in adenocarcinomas. Accordingly, blood group-based specific immuno-oncotherapy could be considered in some cancer cases.

相差镜检联合UF-1000i RBC参数对肾小球性血尿的诊断价值

目的通过分析尿液流式细胞分析仪UF-1000i RBC激光参数与相差镜检对尿红细胞形态判断结果的相关性,探讨RBC激光参数联合相差镜检对肾小球性血尿的诊断价值。方法对120份新鲜尿标本采用日本Sysmex UF-1000i全自动尿沉渣分析仪分析RBC激光参数,同时采用相差显微镜镜检尿红细胞形态,以临床诊疗和病理结果为标准,统计临床诊疗符合率,比较镜检与RBC激光参数的相关性。结果 120例血尿标本中,相差显微镜与尿沉渣分析仪RBC-info对肾小球性血尿诊疗符合率分别是75%和76.7%;ROC曲线分析,RBC参数联合相差显微镜检测对肾小球性血尿诊断效能最高(AUC=0.942)。结论联合应用流式尿沉渣分析仪和相差显微镜检查可提高肾小球性血尿红细胞鉴别能力。

甘油果糖对人RBC、大鼠RBC的作用及输液安全性的研究

目的:探讨甘油果糖注射液(Glycerol and Fructose Injection)对人红细胞(Red blood cell,RBC),大鼠RBC的作用及对大鼠输液安全性的研究,以指导临床应用。方法:以40例健康成人RBC、32只sD大鼠RBC为研究对象,进行体外溶血试验;以72只SD大鼠为研究对象,进行输液安全性研究,研究组为甘油果糖注射液,对照组为生理盐水注射液,应用改良酸化甘油溶解实验测定甘油果糖注射液对人RBC和大鼠RBC的溶解程度,应用邻-甲联苯胺氧化显色法测定甘油果糖注射液对上清液游离血红蛋白(Hemoglbin,HB)含量的影响。同时测定研究组和对照组大鼠血浆游离HB、蛋白非结合胆红素(间接胆红素)、尿素氮(Blood urea nitrogen,BUN)、肌酐的含量,以对甘油果糖注射液安全性进行研究。结果:体外试验为甘油果糖注射液对人RBC和大鼠RBC的溶解程度与生理盐水相似,对上清液游离HB含量无明显影响。对大鼠给予甘油果糖注射液和生理盐水,最快输液速度6ml/min和最大输液量30ml/kg/d,24小时后测定大鼠血浆游离HB、蛋白非结合胆红素、BUN、肌酐的含量,两组结果相似。结论:甘油果糖注射液在体外对人RBC和大鼠RBC无明显溶解作用。在大鼠体内也没有发现有溶血作用,对肾脏功能影响与生理盐水相似。

穴位埋线联合丹莪妇康煎膏治疗子宫内膜异位症痛经疗效及对患者血清CA125的影响

目的:探究采用穴位埋线联合丹莪妇康煎膏疗法对子宫内膜异位症(EMs)痛经患者(气滞血瘀型)的临床治疗效果及其对血清糖类抗原125(CA125)的影响。方法:将66例EMs痛经患者(气滞血瘀型)随机分为两组,各33例。对照组给予丹莪妇康煎膏治疗,观察组在此基础上辅以穴位埋线疗法,两组均连续治疗3个月经周期。记录两组患者的临床疗效和不良反应;比较治疗前、治疗3个月经周期后经期视觉模拟评分量表(VAS)评分、痛经症状评分、气滞血瘀型痛经的中医证候积分及血清CA125水平变化。结果:观察组总有效率96.77%高于对照组86.67%(P<0.05);治疗3个月经周期后,两组的经期VAS评分、痛经症状评分、气滞血瘀型痛经中医证候积分及血清CA125水平均较治疗前降低,且在上述指标的改善幅度方面,观察组均显著优于对照组(均P<0.05);两组不良反应率比较差异无统计学意义(P>0.05)。结论:穴位埋线联合丹莪妇康煎膏可有效缓解气滞血瘀型EMs痛经及其伴随症状,改善中医证候,且能降低血清CA125水平、抑制异位内膜进展,提高临床疗效。

^(99m)Tc-RBC的X射线光电子能谱研究

９９ｍＴｃ－标记的红细胞（９９ｍＴｃ－ＲＢＣ）是目前最常用的血池显象剂。木工作用体内标记法制备了９９ｍＴｃ－ＲＢＣ，采用Ｘ－射线光电子能谱（ＸＰＳ）对９９ｍＴｃ－ＲＢＣ中Ｔｃ原子的化学状态及其变化进行了测定。９０ｍＴｃ－ＲＢＣ于标记后１ｈ与标记后２ｈ所测ＸＰＳ能谱显示，表明９９ｍＴｃ－ＲＢＣ的稳定性佳。从ＲＢＣ与９９ｍＴｃ成键的情况看，多肽键上带负电荷的羧酸根氧原子与９９ｍＴｃ配合；半胱氨酸带负电荷的硫原子与９９ｍＴｃ配合。从９９ｍＴｃ－ＲＢＣ９９ｍＴｃ３ｄ５／２的Ｅｂ值（其中ｊ＝５／２，Ｅｂ为原子能级中电子的结合能），可以推测９９ｍＴｃ－ＲＢＣ中９９ｍＴｃ的价态低于３价。与此同时，对含、９９Ｔｃ（Ｖ）、９９Ｔｃ（ＩＶ）及９９Ｔｃ（Ⅲ）的化合物进行ＸＰＳ测定，其结果表明：化学位移随着９９Ｔｃ还原价态的降低而降低。实验结果与理论推测相符。

E．coli O86 O-Antigen全保护五糖重复单元的化学简易合成

以5个单糖组分为原料,经过7步,以21%的总产率得到E.coliO86抗原全保护的五糖重复单元.在合成路线中,充分利用糖基化反应的立体选择性原则,结合HClO4-SiO2固体催化剂和"IP"策略,大大提高了合成的效率.整个合成路线设计操作简单,选择性高,消耗低,产率高,可以用于快速高效地合成其它一些具有生物活性的寡糖分子.

单抗HIM_(82)对中性粒细胞ROS损伤RBC的保护作用

目的 研究单抗HIM_(82)(30μg/ml)是否能对PMA激活PMN后产生的ROS进行降调节,进而降低ROS对红细胞的损伤,验证HIM_(82)对RBC损伤有无保护作用。方法 用PMA激活PMN后产生的ROS对无HIM_(82)和加HIM_(82)保护的RBC进行损伤实验。结果 加HIM_(82)保护后的RBC膜蛋白乙酰胆碱酯酶(AchE)活性由损伤时的(71.36±22.15)%恢复到(87.60±14.78)%(n=6 P<0.001),活性提高了16.30%;对RBC膜脂质过氧化损伤的保护作用使损伤率从(52.42±20.63)%减小到(23.81±12.40)%(n=9 P<0.01),脂质过氧化损伤降低了54.57%,RBC膜脆性损伤用溶血率表示,溶血率由(80.76±20.23)%下降到(66.30±7.08)%(n=6 P<0.001),下降了17.90%。结论单抗HIM_(82)不仅能明显地降低PMN的ROS水平也能有效保护周围红细胞,可能对防治多种趋化因子激活PMN引起的ROS损伤提供一种新的有效途径。

Anti-HBc screening in Indian blood donors:Still an unresolved issue

AIM:To study the seroprevalence of antibody to hepatitis B core antigen (anti-HBc) in healthy blood donors negative for HBsAg and to evaluate whether anti-HBc detection could be adopted in India as a screening assay for HBV in addition to HBsAg. METHODS: A total of 1700 serum samples collected from HBsAg-negative healthy blood donors were tested for the presence of anti-HBc antibody (IgM + IgG). All samples reactive for anti-HBc antibody were then investigated for presence of anti-HBs and for liver function tests (LFTs). One hundred serum samples reactive for anti-HBc were tested for HBV DNA by PCR method. RESULTS: Out of 1700 samples tested, 142 (8.4%) blood samples were found to be reactive for anti-HBc. It was signif icantly lower in voluntary (6.9%) as compared to replacement donors (10.4%, P = 0.011). Seventy- two (50.7%) anti-HBc reactive samples were also reactive for anti-HBs with levels > 10 mIU/mL and 70 (49.3%) samples were non-reactive for anti-HBs, these units were labeled as anti-HBc-only. These 142 anti-HBc reactive units were also tested for liver function test. HBV DNA was detected in only 1 of 100 samples tested. CONCLUSION: Keeping in view that 8%-18% of donor population in India is anti-HBc reactive, inclusion of anti- HBc testing will lead to high discard rate. Anti-HBs as proposed previously does not seem to predict clearance of the virus. Cost effectiveness of introducing universalanti-HBc screening and discarding large number of blood units versus considering ID NAT (Individual donor nuclic acid testing) needs to be assessed.

Transplantation of microencapsulated umbilical-cord-blood-derived hepatic-like cells for treatment of hepatic failure

AIM: To investigate intraperitoneal transplantation of microencapsulated hepatic-like cells from human umbilical cord blood for treatment of hepatic failure in rats. METHODS: CD34+ cells in umbilical cord blood cells were isolated by magnetic cell sorting. In the in vitro experiment, sorted CD34+ cells were amplified and induced into hepatic-like cells by culturing with a combination of fibroblast growth factor 4 and hepatocyte growth factor. Cultures without growth factor addition served as controls. mRNA and protein levels for hepatic-like cells were analyzed by reverse transcription-polymerase chain reaction, immunohistochemistry and immunofluorescence. In the in vivo experiment, the hepatic-like cells were encapsulated and transplanted into the abdominal cavity of acute hepatic failure (AHF) rats at 48 h after D-galactosamine induction of acute hepatic failure. Transplantation with PBS and unencapsulated hepatic-like cells served as controls. The mortality rate, hepatic pathological changes and serum biochemical indexes were determined. The morphology and structure of microcapsules in the greater omentum were observed. RESULTS: Human albumin, alpha-fetoprotein and GATA-4 mRNA and albumin protein positive cells were found among cultured cells after 16 d. Albumin level in culture medium was significantly increased after culturing with growth factors in comparison with culturing without growth factor addition (P < 0.01). Compared with the unencapsulated group, the mortality rate of the encapsulated hepatic-like cell-transplanted group was significantly lower (P < 0.05). Serum biochemical parameters, alanine aminotransferase, aspartate aminotransferase and total bilirubin in the encapsulated group were significantly improvement compared with the PBS control group (P < 0.01). Pathological staining further supported these findings. At 1-2 wk post-transplantation, free microcapsules with a round clear structure and a smooth surface were observed in peritoneal lavage fluid, surviving cells inside microcapsules were found by trypan blue staining, but some fibrous tissue around microcapsules was also detected in the greater omentum of encapsulated group by hematoxylin and eosin staining. CONCLUSION: Transplantation of microencapsulated hepatic-like cells derived from umbilical cord blood cells could preliminarily alleviate the symptoms of AHF rats.

Study on the packed volume and the void ratio of idealized human red blood cells using a ?nite-discrete element method

Numerical simulations are performed to examine the packing behavior of human red blood cells(RBCs). A combined ?nite-discrete element method(FDEM) is utilized, in which the RBCs are modeled as no-friction and no-adhesion solid bodies. The packed volume and the void ratio of a large number of randomly packed RBCs are clari?ed,and the effects of the RBC shape, the mesh size, the cell number, and the container size are investigated. The results show that the packed human RBCs with normal shape have a void ratio of 28.45%, which is slightly higher than that of the ?at or thick cells used in this study. Such information is bene?cial to the further understanding on the geometric features of human RBCs and the research on RBC simulations.

急诊手术RBC输注量的变化趋势分析

目的分析我院近3年急诊手术异体红细胞(red blood cells,RBC)输注量的变化趋势。方法利用手术麻醉信息系统收集2012年8月至2015年7月共3年度的急诊手术患者,分别统计各年的急诊手术例数、输血人次、异体RBC及自体RBC月均输注量,计算输血率、人均异体RBC及自体RBC输注量并分析其变化趋势。结果 3年度共有16413例急诊手术患者,分别为5300例、5516例、5597例;输血人次逐年减少,分别为231人次、214人次、109人次;输血率比较,第3年度的(1.99%)低于第1年度的(4.36%)及第2年度的(3.88%)(P<0.05),第2年度的(3.88%)低于第1年度的(4.36%)(P<0.05);异体RBC月均输注量比较,第3年度的(52.8±37.8)U低于第一年度的(120.6±35.1)U及第2年度的(85.6±32.1)U(P<0.05),第2年度的(85.6±32.2)U低于第1年度的(120.6±35.1)U(P<0.05);各年度异体RBC人均输注量分别为(6.8±1.9、4.4±0.9、5.4±2.4)U;自体RBC月均输注量分别是(11.8±3.4、16.4±7.5、10.1±7.3)U,自体RBC人均输注量分别是(0.9±0.6、0.9±0.4、1.2±1.1)U,3年度比较,这3个指标变化趋势不明显,差异无统计学意义(P>0.05)。结论输血人次及输血率逐年下降,异体RBC月均输注量逐年减少,但输血患者的异体RBC人均输注量、自体RBC月均输注量及人均自体RBC输注量的变化趋势不明显。