Immunophenotyping of hematopoietic progenitor cells: Comparison between cord blood and adult mobilized blood grafts

AIM:To study the immunophenotype of hematopoietic progenitor cells from cord blood (CB) grafts (n = 39) in comparison with adult apheresis grafts (AG, n = 229) and pre-apheresis peripheral blood (PAPB) samples (n = 908) using flow cytometry analysis.METHODS: First, we performed a qualitative analysis of CD34+ cell sub-populations in both CB and PAPB grafts using the standardized ISHAGE protocol and a wide panel of 20 monoclonal antibodies. Next, we stud-ied some parameters, such as the age of mothers and the weight of newborns, which can influence the qual-ity and the quantity of CD34+ cells from CB. RESULTS: We found that the percentage of apoptotic cells was high in CB in comparison to PAPB (PAPB: 4.6% ± 2.6% vs CB: 53.4% ± 5.2%, P < 0.001). In CB, the weight of newborn and the age of the mother have the influence on CD34+ cells. The follow-up of Ag CD133in the ISHAGE double platform protocol in association with CD45, CD34 and the 7’AAD shows an equal rate between the two cell populations CD133+CD45+CD34+ high and CD34+CD45+ high with a higher percentage. So, is the inclusion of Ac CD133 necessary in the pres-ent panel included in the ISHAGE methodflLast part, we showed a signif icant presence of interferon γ in CB in comparison to PAPB, the annexin showing the high number of apoptotic cells in CB. CONCLUSION: This study demonstrates that many different obstetric factors must be taken into account when processing and cryo-banking umbilical CB units for transplantation.     

Optimization and performance validation for the fully automatic blood type analysis system

To evaluate and validate the application of fully automatic blood type analysis system parameters under actual lab conditions. All key system parameters were optimized and validated accordingly. The optimized parameters were centrifugal speed at 550 rpm; centrifugal time 20 min; resuspension speed 1,200 rpm; resuspension time: 45 s; incubation temperature 25℃ ; incubation time 400 s; incubation rate: 0 rpm. The sampled red blood cell concentration was 3%. The ratio of plasma to red blood cells reagent was 60 μL:30 μL; the ratio of antibody (reagent) to sampled diluted red blood cell was 30 μL:30 μL. After applying our key parameters for optimization and validation, the automatic blood type detection system's performance was found to meet the relevant requirements, effectively improving the accuracy and reliability of the detection system.     

Serological and genetic analysis of a rare CisAB01/O01 blood group

The paper aims to analyze a rare blood sample in Ganzhou City Hospital with CisAB subtype and explore a feasible pattern for blood typing of rare blood type patients, so as to ensure clinical transfusion safety. The routine serological methods were used for ABO forward and reverse blood typing and the fluorescence real-time PCR technique was used for sample genotyping. A human ABO blood group 6-7 exon sequencing kit was used for sequence analysis. The nucleic acid sequence of the sample was compared with reference sequences. The forward typing results demonstrated that the sample was ABw, RhD positive. The sample exhibited 4+ agglutination with anti-H and anti-AB antibodies. Reverse typing by microcolumn gel method showed an AB result, but the serum sample demonstrated weak agglutination with B cell under room temperature, 4 °C and 37 °C in saline when tested with tube method respectively. The serological results matched with the A₂B₃ serotype. The fluorescent real-time PCR genotyping results displayed A/O01. The sequence analysis demonstrated deletion of guanine in 261-position 467C>T (heterozygote) and 803G>T (heterozygote) mutation respectively. The mutation caused the A glycosyltransferase peptide chain to change from proline to leucine (P156L) at 156 and from glutamate to alanine (G268A) at 268. The result demonstrated that the sample''s genotype was CisAB01/O01. The mutation of glycosyltransferase coding gene leads to an abnormal serological reaction pattern. Only by combining the results of genetic analysis can we get the true sample blood type and better ensure the safety of clinical blood transfusion.     

From stem cells to red blood cells:how far away from the clinical application?

The generation of red blood cells(RBCs)from stem cells provides a solution for deficiencies in blood transfusion.Currently,primary hematopoietic stem cells,embryonic stem cells and induced pluripotent stem cells have shown the potential to produce fully mature RBCs.Here,we discuss the advantages,induction protocols,progress and possible clinical applications of stem cells in RBC production.     

An establishment of the procedures to eliminate incorrect blood due to non-matched or mislabeled tubes

This study aims to establish a set of procedures to eliminate incorrect blood due to non-matched tubes or mislabeled tubes. The errors such as these are suspected to have occurred when upon first and second-detection. Under the identical laboratory conditions, the results of re-detection of blood bag samples and rare type antigen samples, as well as re-collected samples from donor, and plasma diluted are not consistent with the original results. Here, 20 antigen erythrocytes were detected for blood bag samples and original samples, in which no incorrect blood was mistakenly administered or mislabeled. Samples taken under identical laboratory conditions were not found to have incorrect blood administered to a non-matched tube. The results showed nonlinear changes by HBsAg ELSIA after plasma dilution. The study suggests that the second-detection taken shortly after first detection is the most appropriate method to detect errors at the earliest time point. Blood bag samples are identical to those of the original antigen identification group, which means that the probability of the samples coming from the same donor is extremely highly. At same time, space and plasma diluted, re-detection can effectively exclude incorrect blood being added to a non-matched tube.     

α_(1A)-adrenergic receptor mediated pressor response to phenylephrine in anesthetized rat

To determine which subtype of α1-adrenergic receptors plays a role in the regulation of blood pressure, with α1A-adrenergic receptor-mediated vasoconstriction in perfused hindlimb as a control, we compared the inhibitory effects of various ai-adrenergic receptor selective antagonists on the vasopressure responses to phenylephrine between the mean arterial pressure and hindlimb perfusion pressure in anesthetized rats. In Normotensive Wistar rats, the results showed that the inhibitory effects (dose ratios of ED50, Dr) of α1-adrenoceptor selective antagonist (prazosin, Dr 13.5±3.6 vs.15.1±4.3, n = 11), α1A-adrenoceptor selective antagonist (5-methyl-urapidil, Dr 2.4±0.9 vs. 3.7±2.3, n = 12; RS-17053, Dr 3.2±1.6 vs. 4.4±3.3, n =12) and α1D-adrenoceptor selective antagonist (BMY7378, Dr 1.9±0.9 vs. 2.2±0.8, n = 8) on phenylephrine- induced increases of perfusion pressure in the autoperfused femoral beds were the same as that in the mean arterial blood pressure in normotensive Wistar rats. The i     

Absent MicroRNAs in Different Tissues of Patients with Acquired Cardiomyopathy

MicroRNAs (miRNAs) can be found in a wide range of tissues and body fluids, and their specific signatures can be used to determine diseases or predict clinical courses. The miRNA profiles in biological samples (tissue, serum, peripheral blood mononuclear cells or other body fluids) differ significantly even in the same patient and therefore have their own specificity for the presented con-dition. Complex profiles of deregulated miRNAs are of high interest, whereas the importance of non-expressed miRNAs was ignored. Since miRNAs regulate gene expression rather negatively, absent miRNAs could indicate genes with unaltered expression that therefore are normally expressed in specific compartments or under specific disease situations. For the first time, non-detectable miRNAs in different tissues and body fluids from patients with different diseases (cardiomyopathies, Alzheimer’s disease, bladder cancer, and ocular cancer) were analyzed and com-pared in this study. miRNA expression data were generated by microarray or TaqMan PCR-based platforms. Lists of absent miRNAs of primarily cardiac patients (myocardium, blood cells, and serum) were clustered and analyzed for potentially involved pathways using two prediction platforms, i.e., miRNA enrichment analysis and annotation tool (miEAA) and DIANA miRPath. Extensive search in biomedical publication databases for the relevance of non-expressed miRNAs in predicted pathways revealed no evidence for their involvement in heart-related pathways as indicated by software tools, confirming proposed approach.     

ABO blood group system

The ABO blood group system in humans has three different carbohydrate antigens named A, B, and O. The A antigen sequence is terminal trisaccharide N-acetylgalactosamine (GalNAc)α1-3[Fucα1-2]Galβ-, B is terminal trisaccharide Galα1-3[Fucα1-2]Galβ-, and O is terminal disaccharide Fucα1-2Galβ-. The single ABO gene locus has three alleles types A, B and O. The A and B genes code A and B glycosyltransferases respectively and O encodes an inactive enzyme. A large allelic diversity has been found for A and B transferases resulting in the genetic subgrouping of each ABO blood type. Genes for both transferases have been cloned and the 3D structure of enzymes with and without substrate has been revealed by NMR and X ray crystallography. The ABO blood group system plays a vital role in transfusion, organ and tissue transplantation, as well as in cellular or molecular therapies.     

Serological?identification and molecular?characterization?of?B?(A)?02?subtype?in?patients?and blood donors from?Eastern?China

This study was designed to identify the rare type ABO blood groups, B(A) 02, from Eastern China. Three samples with discordant serological results during routine blood type identification and four samples from one sample’ family were selected. All of them were detected by serological method. The exon 6 and 7 of the ABO genes were amplified by PCR and sequenced. They were typed as AsubB by serology and as BO by genotype. In AsubB samples, nt 700C>G mutation was detected in B gene, which was previously defined as B(A)02 alleles. In these seven samples, six showed B(A)02/O01 and one showed B(A)02/O02.B(A)02 allele was found to be more common in this study than B(A)04 which is considered to be more frequent than B(A)02. The careful identification of rare blood types is important for the safety of clinical blood transfusion.     

Comparing gene expression profiles of Kashin-Beck and Keshan diseases occurring within the same endemic areas of China

In this study, differentially expressed genes in peripheral blood from patients with Kashin-Beck disease and Keshan disease were compared to further investigate the etiology and pathogenesis of both diseases, which occur in a common endemic area of China. Twenty Kashin-Beck disease patients and 12 healthy controls, and 16 Keshan disease patients and 16 healthy controls, were grouped into four pairs. Patients and controls were selected from common endemic areas for the two diseases. Total RNA was isolated from peripheral blood mononuclear cells from all patients and controls, and gene expression profiles analyzed by oligonucleotide microarrays. Sixteen genes differentially expressed in both Kashin-Beck disease and Keshan disease (versus controls) were identified, and comprised nine genes showing synchronous and seven asynchronous expression. The Comparative Toxicogenomics Database shows that expression and biological function of these genes can be affected by multiple environmental factors, including mycotoxin and selenium content, potential environmental risk factors for the two diseases. Thus, these shared differentially expressed genes may contribute to the distinct organ lesions, caused by common environmental risk factors of Kashin-Beck disease and Keshan disease.     

Quantitative assessment of acute kidney injury by noninvasive arterial spin labeling perfusion MRI:a pilot study

The kidneys are essential for maintaining homeostasis,are responsible for the reabsorption of water,glucose and amino acids,and filter the blood by removing waste.Acute kidney injury(AKI) is a syndrome characterized by the rapid loss of renal excretory function and the accumulation of end metabolic products of urea and creatinine.AKI is associated with the later development of chronic kidney disease and end-stage kidney disease,and may eventually be fatal.Early diagnosis of AKI and assessments of the effects of treatment,however,are challenging.The pathophysiological mechanism of AKI is thought to be the imbalance between oxygen supply and demand in the kidneys.We have assessed the ability of arterial spin labeling(ASL) perfusion magnetic resonance imaging(MRI),without the administration of contrast media,to quantify renal blood flow(RBF) non-invasively.We found that RBF was significantly lower in AKI patients than in healthy volunteers.These results suggest that ASL perfusion MRI,a noninvasive measurement of RBF,may be useful in the early diagnosis of AKI.     

德昂族红细胞血型分布的研究

对100名德昂族人的红细胞血型进行了调查。结果表明,ABO系统中r>p>q; MNSs系统中m>n,s>S,Ms>Ns>MS>NS;Rh系统中CCDee表型最常见,CDe单倍型频率较高(0.8250),而cDe频率(0.0352)较低;P系统中P1基因频率为0.1 340。德昂族的红细胞血型分布具有我国南方民族的特点。 Abstract:A total number of 100 unrclated individuals of De’ang ethnic group in Luxi County,Yunnan Province were examined for the distribution of ABO,MNSs,Rh and P blood groups.The gene frequencies were as follows:r=0.6580,p=0.1740,q=0.1680,m=0.6500,n=0.3500,S=0.0200,s=0.9800,MS=0.0200,Ms=0.6300,NS=0,Ns=0.3500,C=0.8250,D=1.0000,E=0.1398,Cde=0.8250,cDE=0,1387,cDe=0.0352,P1=0.1340.At the same time,no MS,MNS,NS,NSs,CCDE and ccDee phenotype were found.These results show that the distribution of these systems in De’ang ethnic group has the characteristics of the nationalities living in South China.     

南瓜多糖的降血脂作用研究

研究了南瓜多糖对正常小鼠和四氧嘧啶型糖尿病小鼠血脂的影响.对正常小鼠灌胃高低两个剂量(200、400mg/kg)的南瓜多糖水溶液;对建模成功的小鼠随机分为模型对照组、南瓜多糖组(400mg/kg的PP)和优降糖组(15mg/kg的优降糖),灌胃给药.对两种类型小鼠的处理均设立正常对照组,给予等体积的生理盐水,测定小鼠的TC、 TG、 LDL-C、 HDL-C的含量.结果表明高剂量的南瓜多糖对正常小鼠的降血脂效果优于低剂量;南瓜多糖使糖尿病小鼠的TC、 TG、 LDL-C显著降低,HDL-C极显著升高;并且南瓜多糖的降血脂效果与优降糖相比无显著差异.     

溴氰菊酯连续暴露对罗非鱼血清乙酰胆碱酯酶活性的影响

以罗非鱼为受试生物,研究了不同水温(23℃～27 ℃)对罗非鱼血清中乙酰胆碱酯酶活性的影响,并在此基础上研究了不同浓度(1.0、2.0、3.0、5.0和10.0 μg·L-1)溴氰菊酯暴露下,罗非鱼血清中乙酰胆碱酯酶活性的动态变化.水温分别为23℃和27℃时,乙酰胆碱酯酶的活性分别为(2.75±0.21)和(2.73±0.26)U·ml-1,活性波动范围分别为-12.0%～13.1%和-11.0%～14.2%.水温为(25±1)℃,染毒10 d时,2.0μg·L-1以上浓度的溴氰菊酯对罗非鱼血清中乙酰胆碱酯酶具有明显的抑制作用;染毒20 d时,2.0μg·L-1以上浓度的溴氰菊酯对其抑制率均超过40%;染毒25 d时,5.0 μg·L-1的溴氰菊酯对其抑制率达到最大,为62.3%.实验结果表明:水温在23℃～27℃的波动不会对罗非鱼血清中乙酰胆碱酯酶的活性产生显著影响;水温为(25±1)℃时,高浓度溴氰菊酯(≥2.0 μg·L-1)会对罗非鱼血清中乙酰胆碱酯酶的活性产生抑制作用,而且抑制率随染毒时间的延长呈增加趋势.     

HBV感染者外周血TLR4mRNA表达与疾病进展关系研究

构建pUCm-TLR4质粒作为定量模板,采用实时荧光定量RT-PCR的方法检测 140 例HBV感染者外周血单个核细胞(PBMC)中TLR4mRNA的含量,并结合HBV感染者临床情况进行分析.HBV感染者PBMC中TLR4mRNA的表达范围是1.1×105～10.5×107 copy/μg RNA,显著高于健康对照组(4.5×105～1.25×106 copy/μg RNA),其中慢性乙肝组TLR4mRNA的表达明显高于慢性重型乙肝组(P＜0.05);HBV病毒载量的对数值与TLR4mRNA的含量存在负相关(r=-0.316,P＜0.01).结果表明,HBV感染者PBMC中TLR4mRNA的表达与疾病进展明显相关.     

样品预处理对HCV-cAg酶联免疫吸附试验特异性的影响

目的:探讨血液样品预处理后对控制酶联免疫吸附试验中假阳性的效果.方法:向稀释血清样品中加入适量的C2H6OS和IgM抗体后,以HCV-cAg酶联免疫方法检测60份血清样品中丙型肝炎病毒核心抗原的阳性情况;与常规样品处理方法和逆转录-聚合酶链反应(RT-PCR)检测结果为标准进行比较分析.结果:60份血清样品,采用常规的样品处理方法检测出的阳性例数为34例,假阳性率迭25%;采用改良法检测出的阳性例数为23例,假阳性率达6.7%,二者相比差别具有统计学意义(P(≦) 0.05).结论:改良法血清样品预处理可提高ELISA法检出丙型肝炎病的特异性.     

血液净化治疗毒鼠强急性中毒的临床研究

目的 观察血液净化治疗对毒鼠强急性中毒的疗效.方法 在内科常规治疗的基础上,采用血液净化救治急性毒鼠强中毒10例,并与单纯常规治疗的12例对照,比较两组的平均住院天数、治愈率以及APACHE Ⅲ评分变化.结果 血液净化治疗组平均住院天数较常规治疗组明显缩短(P＜0.05),治愈率明显提高(P＜0.05);两组治疗后APACHEⅢ评分均较治疗前降低,其中血液净化组治疗后APACHEⅢ评分降低尤为明显(P＜0.05).结论 血液净化是救治毒鼠强急性中毒的有效方法.     

解除模拟失重后清醒大鼠血压信号的谱分析

本文旨在观察模拟失重28 d大鼠解除尾部悬吊前、后(2 h内),清醒自由活动状态下动脉收缩压(systolic bloodpressure,SBP)、舒张压(diastolic blood pressure,DBP)和心率(heart rate,HR)的变化.采用自回归模型法对不同时间点的收缩压变异性(SBP variability,SBPV)和心率变异性(HR variability,HRV)进行自谱与互谱分析,并比较自回归法与周期图法的自谱分析结果:由传递函数分析得到反映压力感受器-心率反射敏感性(baroreceptor-heart rate reflex sensitivity,BRS)变化的有关数据.结果显示,用自回归模型法对清醒大鼠血压信号进行短时程谱分析可得到较为平滑、谱峰清楚的谱估计曲线.28 d模拟失重大鼠解除尾部悬吊前、后,SBP、DBP和HR及其主要谱指标,以及高、低频段传递函数的平均增益均无显著性变化,不同时间点的谱指标也无显著差别;但模拟失重组的SBP、DBP和HR却显著高于对照组.上述结果提示,中期模拟失重大鼠恢复正常体位后,其清醒状态的BPV与HR均处于升高状态,但其短时程BPV与HRV谱及BRS均无显著变化,与最近报道的航天员HRV与BRS无显著改变一致.     

原发性肝癌:CT血液供应、P53抗体和血管内皮生长因子

目的:分析原发性肝癌(primary hepatic carcinoma,简称肝癌)血液供应(blood supply,简称血供)CT类型与患者血清P53抗体、血管内皮生长因子(vascular endothelial growth factor,VEGF)的关系,以及血清P53抗体与VEGF的相关性.方法:用酶联免疫吸附法定量检测首诊肝癌患者血清P53抗体和VEGF,与CT诊断的血供类型结果进行对比,分析不同血供类型的肝癌是否存在血清P53抗体差异(单因素分析),并分析不同血供类型的肝癌血清VEGF是否存在差异(单因素分析),然后分析血清P53抗体和VEGF之间的相关性.结果:四种不同血供类型的肝癌之间血清P53抗体滴度有差异(P<0.05).值得注意的是,富血供型与混合型之间、动-静脉漏型(AVS)与混合型之间血清P53抗体有显著差异(P<0.05),富血供型与AVS型之间血清P53抗体没有显著差异(P>0.05).不同血供类型的肝癌中,富血供型和混合型血清P53抗体与VEGF呈正相关(P<0.05),而AVS型和乏血供型中,血清P53抗体与VEGF不呈直线相关(P>0.05).结论:血清P53抗体和血清VEGF滴度在血供不同肝癌中滴度有差异,在富血供型和混合型中血清P53抗体与VEGF滴度呈正相关.     

中国栅蛛属2新种记述(蜘蛛目:栅蛛科)

本文记了分别采自云南高黎贡山的栅蛛科栅蛛属Hahnia 2新种:垭口栅蛛,新种S.yakouensis sp.nov.和肾形栅蛛,新种S.reniformis sp.nov..垭口栅蛛后眼列前曲,交媾腔大,扁圆形,交媾孔1个,位于交媾腔下缘,交媾管粗,呈"人"字形下行分成2支再向两侧扭曲.纳精囊有一肓管斜向上伸出,鉴于上述特征而与Hahnia mridulae Tikader,1970不同.肾形栅蛛交媾孔2个,位于生殖厣腹面中央,纳精囊1对,大,肾形,插入器始于生殖球左下方,鉴于上述特征而与Hahnia xinjiangensis Wang et Liang,1989不同. Abstract： The present paper deals with two new species of the genus Hahnia collected from the Gaoligong Mountains Region of Yunnan Province, China: Hahnia yakouensis sp. nov., Hahnia reniformis sp. nov..