Strong linkage disequilibrium of a HbE variant with the (AT)9(T)5 repeat in the BP1 binding site upstream of the β-globin gene in the Thai population

A binding site for the repressor protein BP1, which contains a tandem (AT)_x(T)_y repeat, is located approximately 530 bp 5 to the human -globin gene (HBB). There is accumulating evidence that BP1 binds to the (AT)₉(T)₅ allele more strongly than to other alleles, thereby reducing the expression of HBB. In this study, we investigated polymorphisms in the (AT)_x(T)_y repeat in 57 individuals living in Thailand, including three homozygotes for the hemoglobin E variant (HbE; 26Glu->Lys), 22 heterozygotes, and 32 normal homozygotes. We found that (AT)₉(T)₅ and (AT)₇(T)₇ alleles were predominant in the studied population and that the HbE variant is in strong linkage disequilibrium with the (AT)₉(T)₅ allele, which can explain why the ^E chain is inefficiently synthesized compared to the normal ^A chain. Moreover, the mildness of the HbE disease compared to other hemoglobinopathies in Thai may be due, in part, to the presence of the (AT)₉(T)₅ repeat on the HbE chromosome. In addition, a novel (AC)_n polymorphism adjacent to the (AT)_x(T)_y repeat (i.e., (AC)₃(AT)₇(T)₅) was found through the variation screening in this study.MIM and accession numbers and URLs for data presented herein are as follows: Online Mendelian Inheritance of Man (OMIM), (for HBB [MIM 141900]). GenBank, (accession number [NG_000007.2] reference sequence information).     

重组人类杀菌肽基因在CHO细胞中的表达

目的 检测重组人类杀菌肽基因（BPI）在真核细胞中的表达.方法 采用脂质体转染法将本室构建的pCDNA3.1-BP1500及pCDNA3.1-BPI600重组子导入中国仓鼠卵巢上皮细胞（CHO）,G418筛选出带有重组子的细胞集落,RT-pCR检测BPI在CHO细胞集落中的表达.结果 转染后G418抗性CHO细胞中可检测到BPImRNA的表达.结论 pCDNA3.1-BPI重组子被成功转染至CHO,并得到有效表达.     

Therapeutic potential of the

Innate immune mechanisms respond rapidly to bacterial infection. A key cellular component of the innate immune response is the neutrophil, whose cytoplasmic granules contain a variety of antimicrobial proteins and peptides. Among these is the bactericidal/permeability-increasing protein (BPI), a cationic 55 kDa protein whose selective anti-infective action against Gram-negative bacteria is based on its high (nM) affinity for lipopolysaccharide (LPS, or “endotoxin”). Binding of BPI to Gram-negative bacteria results in growth inhibition, serves as an opsonin that enhances phagocytosis of bacteria and inhibits bacteria-induced inflammatory responses by blocking the interaction of LPS with host pro-inflammatory pathways. Expression of BPI appears to be developmentally regulated as human newborns apparently have lower neutrophil BPI levels than adults. BPI expression has also recently been demonstrated in human epithelial cells where it appears to be inducible by endogenous anti-inflammatory lipids (lipoxins). BPI’s potent anti-endotoxic activity against a broad range of Gram-negative bacterial pathogens is manifest in biological fluids and renders it an attractive template for pharmaceutical development. Indeed, rBPI₂₁, an active recombinant protein derived from human BPI, has proven safe in Phase I human trials, shown promise in Phase II trials and has recently completed a Phase III trial for severe meningococcaemia with apparent benefit. Identification and evaluation of additional disease entities characterised by Gram-negative bacteraemia and/or endotoxaemia as possible targets for BPI therapy continues.     

160例恶性肿瘤患者疼痛负担及其影响因素研究

目的：探索中医药治疗恶性肿瘤患者的疼痛负担及其影响因素。方法：选取2019年9月至2021年12月中国中医科学院广安门医院等10家中医药三甲医院初次就诊的恶性肿瘤患者临床数据,运用简明疼痛调查量表(BPI)进行疼痛评估,采用多因素回归分析方法探讨疼痛负担的影响因素。结果：160例近期未进行抗肿瘤治疗的恶性肿瘤患者中,62.5%为男性患者,65岁以上的患者为55.6%,75.6%临床分期为Ⅳ期,80.6%患者正在使用阿片类药物;癌症患者在过去24 h疼痛平均程度为4分,疼痛最剧烈程度为6分;在疼痛对功能的干扰方面,总体干扰平均得分为4.94分,活动干扰及情感干扰得分分别为5.11分、4.69分;多因素分析中,正在服用阿片类药物的患者具有更高的疼痛最剧烈程度(β=0.239,95%CI为0.267～1.285,P=0.003)。结论：寻求中医药治疗的恶性肿瘤患者疼痛负担总体处于轻至中度水平,疼痛对患者总体功能、活动、情感干扰产生中度影响;正在服用阿片类药物的患者出现爆发痛时疼痛程度更高。     

The effect of enzyme replacement therapy on clinical outcomes in female patients with Fabry disease – A systematic literature review by a European panel of experts

BackgroundHeterozygous females with Fabry disease have a wide range of clinical phenotypes depending on the nature of their mutation and their X-chromosome inactivation pattern; it is therefore important to examine outcomes of enzyme replacement therapy (ERT) in the female patient population specifically. This paper presents the findings of a systematic literature review of treatment outcomes with ERT in adult female patients.MethodsA comprehensive systematic literature review was conducted through January 2017 to retrieve published papers with original data on ERT in the treatment of Fabry disease. The review included all original articles that presented ERT outcomes data on patients with Fabry disease, irrespective of the study type.ResultsClinical evidence for the efficacy of ERT in female patients was available from 67 publications including six clinical trial publications, and indicates significant reductions in plasma and urine globotriaosylceramide (GL-3) accumulation (in female patients with elevated pre-treatment levels) and improvements in cardiac parameters and quality of life (QoL). To date, data are insufficient to conclude on the effects of ERT on the nervous system, gastrointestinal manifestations, and pain in female patients with Fabry disease.ConclusionsThis review of available literature data demonstrates that ERT in adult female patients with Fabry disease has a beneficial effect on GL-3 levels and cardiac outcomes. The current evidence also suggests that ERT may improve QoL in this patient population, though further studies are needed to examine these results.     

Anti‐neutrophil cytoplasmic antibodies directed against the bactericidal/permeability‐increasing protein (BPI) in pediatric cystic fibrosis patients do not recognize N‐terminal regions important for the anti‐microbial and lipopolysaccharide‐binding activity of BPI

This study was performed to examine the prevalence and clinical correlates of bactericidal/permeability‐increasing protein anti‐neutrophil cytoplasmic antibodies (BPI‐ANCA) in pediatric cystic fibrosis (CF) patients and to elucidate their possible role in CF pulmonary pathology. Sera of 27 CF patients were tested for ANCA by indirect immunofluorescence (IFT) and by enzyme‐linked immunosorbent assay (ELISA) for ANCA sub‐specificities. BPI‐ANCA were examined by using standard ELISA for BPI, lipopolysaccharide‐binding protein (LBP), and BPI/LBP fusion proteins to epitope map the main binding sites and look for cross‐reactivity with LBP. Pulmonary function and serum concentrations of total immunoglobulin G (IgG) were measured and infections were diagnosed. In addition, release of reactive oxygen species (ROS) by neutrophil granulocytes was measured after stimulation with monoclonal BPI‐ANCA. Using IFT, two patients showed atypical ANCA staining, six patients exhibited perinuclear ANCA staining, and no cytoplasmic ANCA staining was detected. Of 27 patients, 13 (48%) were BPI‐ANCA (IgG) positive, and three were also immunoglobulin A (IgA) BPI‐ANCA positive; one patient had ANCA against lactoferrin; and no proteinase 3 ANCA was detected in any of the patients. All BPI‐ANCA bound to the C‐terminal region of the molecule; none bound to the N‐terminus or to LBP. There was no significant correlation between clinical data and the occurence of BPI‐ANCA in this cross‐sectional study. Release of ROS from granulocytes was induced by monoclonal BPI‐ANCA. Activation of neutrophils and possible modulation of BPI‐mediated opsonophagocytosis and disposal of Gram‐negative bacteria and lipopolysaccharides by BPI‐ANCA raise the possibility that they contribute to pulmonary pathology in pediatric CF patients but intervention longitudinal studies in large groups of patients are needed to establish a causative association.     

肝硬化患者血清内毒素调节蛋白水平及其临床意义

目的：探讨检测伴肠源性内毒素血症的肝硬化患者血中内毒素结合蛋白(LBP)、杀菌性／通透性增加蛋白(BPI)及可溶性sCD14水平的临床意义。方法：应用基质显色法和ELISA双抗体夹心法，检测45例肝硬化患者血中内毒素脂多糖(LPS)、LBP、BPI和sCD14水平，并以15例健康献血员作为对照。结果：肝功能为A级、B级和C级的肝蘸化患者其血中LPS、LBP、BPI和sCD14水平均明显高于献血员；肝硬化死亡息者的LPS、LBP、BPI和sCD14水平显著高于存活者。结论：肝硬化患者伴肠源性内毒素血症时，血清LBP和sCD14水平升高和BPI相对不足，可显著提高机体对LPS的敏感性。     

Increased bactericidal/permeability increasing protein in patients with cirrhosis

Background: High levels of endotoxin in patients with cirrhosis are thought to be responsible for the activation of tumour necrosis factor‐α (TNF)‐α‐mediated pro‐inflammatory pathways involved in haemodynamic alterations. Bactericidal/permeability increasing protein (BPI) is a protein found in neutrophils with endotoxin‐binding and neutralization capacity. It is not known whether defective BPI production or release is present in cirrhosis. Aims: We investigated the levels of BPI in cirrhotic patients and its relation to other endotoxin‐binding proteins and inflammatory markers. Methods: Plasmatic levels of BPI, lipopolysaccharide‐binding protein, soluble CD14, TNF‐α and BPI mRNA expression in neutrophils were determined in 130 patients and 30 healthy controls. The capacity of patients' plasma to inhibit lipopolysaccharide (LPS)‐mediated TNF‐α production by monocytes from healthy donors was assessed in vitro. Results: Patients with cirrhosis exhibited an increase in BPI mRNA and plasma level of BPI when compared with healthy controls (P<0.05). Child C group displayed the highest frequency of patients with a high concentration of BPI. A positive correlation was found between TNF‐α and plasma levels of BPI (P<0.01). High levels of BPI in plasma were able to significantly reduce in vitro TNF‐α release by monocytes after a challenge with LPS (8.54 ± 1.04 vs. 10.44 ± 0.85 pg/ml, P=0.028). Conclusion: BPI is increased in cirrhotic patients, especially in those with more severe liver disease. The amount of BPI in the plasma correlated with the TNF‐α level and was able to reduce LPS‐mediated TNF production by monocytes. BPI possibly plays a regulatory role by antagonizing the pro‐inflammatory mechanisms mediated by TNF‐α.