中性粒细胞/淋巴细胞比值和血小板/淋巴细胞比值与2型糖尿病患者肾小管损伤的相关性研究

目的探讨中性粒细胞/淋巴细胞(NLR)和血小板/淋巴细胞(PLR)与2型糖尿病(T2DM)患者肾小管损伤的相关性。方法收集2018年9月至2019年9月于中南大学湘雅三医院住院治疗的268例T2DM患者,根据尿白蛋白/肌酐比值(UACR),将其分为正常白蛋白尿组(UACR<30 mg/g)、微量白蛋白尿组(30 mg/g≤UACR<300 mg/g)和大量白蛋白尿组(UACR≥300 mg/g)。收集纳入患者的一般临床资料。空腹抽取肘静脉血检测血常规以及糖化血红蛋白、超敏C反应蛋白(hs-CRP)等生化指标,计算NLR、PLR和预估肾小球滤过率(eGFR),并检测尿α1微球蛋白(α1-MG)、β2-微球蛋白(β2-MG)、视黄醇结合蛋白(RBP)、尿肌酐(Ucr),以α1-MG/Ucr、β2-MG/Ucr和RBP/Ucr作为反映肾小管损伤的指标。采用χ2检验、方差分析及非参数检验对各组指标的差异进行比较,采用Spearman相关分析法分析各炎性指标与肾损伤标志物的相关性,采用受试者工作特征(ROC)曲线分析模型预测价值。结果大量白蛋白尿组(94例)的NLR和PLR明显高于正常白蛋白尿组(94例)和微量白蛋白尿组(80例),微量白蛋白尿组NLR高于正常白蛋白尿组,差异有统计学意义(均P<0.05)。随着NLR、PLR四分位数的递增,患者UACR及RBP/Ucr、α1-MG/Ucr、β2-MG/Ucr逐渐升高,差异具有统计学意义(均P<0.05)。Spearman相关分析显示,NLR、PLR与UACR、RBP/Ucr、α1-MG/Ucr、β2-MG/Ucr均呈正相关(r=0.254~0.385,均P<0.01),与eGFR呈负相关(r=-0.328、-0.151,均P<0.01)。ROC曲线显示,NLR判定UACR>30 mg/g、eGFR<60 ml·min^-1·(1.73 m²)^-1及肾小管损伤的曲线下面积(AUC)分别为0.700、0.679、0.717、0.702、0.737,灵敏度分别为75.29%、85.87%、80.88%、83.74%、85.59%,均优于PLR(AUC分别为0.639、0.639、0.659、0.644、0.676,灵敏度分别为58.05%、75.00%、63.24%、65.04%、58.56%)和hs-CRP(AUC分别为0.597、0.559、0.618、0.644、0.653,灵敏度分别为62.50%、30.12%、63.93%、68.42%、70.87%)。结论NLR和PLR与T2DM患者肾小管损伤密切相关,NLR在综合反映肾小球及肾小管损伤方面优于PLR和hs-CRP。     

肾小管功能在亚临床糖尿病肾病诊断价值中的初步探讨

目的 观察2型糖尿病患者的肾小管功能,分析肾小管损伤标记物与亚临床糖尿病肾病(DN)的关系.方法 298例2型糖尿病患者根据24h尿微量白蛋白(UMA)和估算的肾小球滤过率(GFR)分为对照组(NC组,正常白蛋白尿及正常滤过率,n=100)、亚临床DN组(SDN组,正常白蛋白尿高滤过率,n =87)和早期DN组(EDN组,微量白蛋白尿,n=111).应用免疫透射比浊法检测24 h UMA、尿视黄醇结合蛋白(RBP)、尿β2-微球蛋白(β2-MG),生化法测定尿N-乙酰-D-氨基葡萄糖苷酶(NAG)、β-半乳糖苷酶(GAL),分析各组肾小管损伤标记物及其与亚临床DN的关系.结果 与NC组相比,SDN组NAG较高(x2=9.032,P＜O.01),而与EDN组差异无统计学意义;EDN组RBP水平明显高于NC组及SDN组(x2=56.341,31.955,P均＜0.01);NC组、SDN组、EDN组GAL、β2-MG水平呈升高趋势,但差异无统计学意义.与NC组相比,SDN组及EDN组近端肾小管功能异常率较高(分别为59％,82％,78.2％,x2=42.658,P＜0.01).SDN组肾小管损伤标记物中一项或两项异常者的比例高于NC组及EDN组.肾小管损伤标记物中两项异常者有发生亚临床DN的风险[优势比(OR)=2.9,P=0.005],但经校正年龄、性别、糖尿病病程、总甘油三酯、总胆固醇、高密度脂蛋白-胆固醇、糖化血红蛋白、体重指数后,差异无统计学意义(OR=1.7,P＞0.05).结论 肾小管损伤可能对早期肾功能减退的判断较肾小球损伤更重要,尿白蛋白正常但GFR升高的亚临床DN患者肾小管损伤标记物的水平升高及种类增多.     

尿NAG、血清β-TP水平与糖尿病早期肾病的相关性

目的:探讨2型糖尿病(T2DM)患者尿N-乙酰-β-d-氨基葡萄糖苷酶(NAG)、血清β痕量蛋白(β-TP)水平与糖尿病肾病的相关性。方法:选择2012年1月~2015年3月在邯郸市人民医院内科及河北工程大学附属医院内分泌科住院的T2DM患者130例,根据24h尿白蛋白排泄(UAE)水平,患者被分为尿蛋白正常组(55例),微量蛋白尿组(40例)和大量蛋白尿组(35例)。另选50例同期健康体检者作为健康对照组。测定比较各组尿液NAG含量及血清β-TP水平,并分析其相关性。结果:与健康对照组比较,T2DM患者尿液NAG[(3.4±2.0)U/L比(11.9±3.2)U/L]及血清β-TP水平[(0.53±0.12)mg/L比(0.85±0.24)mg/L]均显著升高(P均0.01);与尿蛋白正常组比较,微量蛋白尿组和大量蛋白尿组尿液NAG[(5.9±3.6)U/L比(9.0±3.9)U/L比(21.0±7.2)U/L]和血清β-TP[(0.64±0.10)mg/L比(0.83±0.23)mg/L比(1.12±0.39)mg/L]水平显著升高,且大量蛋白尿组的显著高于微量蛋白尿组的(P均0.05)。直线相关分析显示,T2DM患者尿液NAG水平与血清β-TP水平呈显著正相关(r=0.756,P=0.029)。结论:尿液NAG含量及血清β-TP水平可反映糖尿病早期肾病的病情,对其早期诊断及疗效判断具有较好的临床应用价值。     

2型糖尿病患者血清护骨素水平与糖尿病肾病的关系

目的 2型糖尿病(T2DM)患者血清护骨素水平与糖尿病肾病(DN)的关系.方法 110例T2DM患者根据尿白蛋白排泄率(UAER)分为正常白蛋白尿组 38例、微量白蛋白尿组 36例及大量白蛋白尿组 36例,同时选择年龄、性别相匹配的40例健康者为对照组.血清护骨素采用ELISA法测定.结果 在T2DM各组中,大量白蛋白尿组患者的血清护骨素水平[(4.45±0.76)ng/L]显著高于微量白蛋白尿组[(3.62±0.70)ng/L]、正常白蛋白尿组[(2.77±0.78)mg/L]和对照组[(2.29±0.37)ng/L](均P＜0.01);微量白蛋白尿组患者的血清护骨素水平高于正常白蛋白尿组.血清护骨素水平与空腹血糖(FPG)、糖化血红蛋白(HbA1c)、超敏C反应蛋白(hs-CRP)及UAER呈明显正相关.血清护骨素水平是DN的重要影响因素.结论 DN患者血清护骨素水平明显升高,护骨素可能在DN的发生中起一定作用.     

糖尿病前期及糖尿病患者尿β2-微球蛋白水平的对比分析

糖尿病肾病(DN)是由糖尿病(DM)微血管病变并发症之一,其早期干预非常重要.但在DM前期和DM早期尿检测多为正常,一旦出现蛋白尿,肾损害亦难以逆转.尿β2-微球蛋白(β2-MG)是早期肾损害的标志之一,它不仅反映了肾小球的滤过率,也是反映近端肾小管功能及渗出性蛋白尿的灵敏指标.国内外目前对DM病人尿微量白蛋白、β2-MG的改变研究较多[1～4],但对演变过程中尿β2-MG的升高起源于何时以及与胰岛素水平、血脂之间的关系报道不多.     

循证医学在2型糖尿病肾病早期诊断中的应用

目的 利用循证检验医学的思维方法,在众多肾病早期筛查的指标中,选出一组判断2型糖尿病(T2DM)早期肾脏损伤的组合.方法 分别检测健康对照组80例和T2DM组287例尿液尿微量白蛋白(mALB)、β2微球蛋白(β2-MG)、α1微球蛋白(α1-MG)、转铁蛋白(TRF)、尿N-乙酰-β-D-氨基葡萄糖苷酶(NAG)水平,按超过健康对照组正常上限(x±2 s),统计阳性例数及阳性率.结果 肾小球、肾小管损伤指标mALB TRF α1-MG NAG或mALB TRF β2-MG NAG联合检测较传统mALB β2-MG联合检测及仅联合检测肾小球或肾小管损伤指标,检出率高.检测mALB TRF α1-MG NAG与mALB TRF β2-MG NAG比较无显著差别.结论 肾小球、肾小管损伤指标联合检测中,建议选择mALB TRF α1-MG NAG或mALB TRF β2-MG NAG联检,影响因素少,结果稳定,检出率高.     

血清胱抑素C、视黄醇结合蛋白与β2-微球蛋白检验在早期2型糖尿病肾病中的应用价值分析

目的 分析血清胱抑素C(cystatin C, Cys C)、视黄醇结合蛋白（retinol binding protein, RBP）与β2-微球蛋白（beta-2-microglobulin, β2-MG）在早期2型糖尿病肾病（diabetic nephropathy, DN）检验中的应用价值。方法 回顾性分析2020年1月—2022年1月期间联勤保障部队第970医院威海医疗区收治的72例早期2型DN患者的临床资料,将其作为研究组;回顾性分析同期本院收治的78例单纯2型糖尿病患者的临床资料,将其作为对照组。两组患者均接受Cys C、RBP、β2-MG及尿微量白蛋白（microalbuminuria, m Alb）检验。比较两组患者的Cys C、RBP、β2-MG及m Alb水平,比较Cys C、RBP、β2-MG单独检验与联合检验2型DN的敏感度、特异度,并分析Cys C、RBP、β2-MG与早期2型DN患者m Alb的相关性。结果 研究组Cys C(1.26±0.12)mg/L、RBP(88.38±12.16)mg/L、β2-MG(3.85±0.50)mg/L、m Alb(90....     

全身炎性反应综合征患儿肾功能相关指标的测定

目的观察血尿有关指标测定与全身炎性反应综合征（SIRS）患儿早期肾脏功能改变的关系。方法对48例SIRS早期患儿进行血清β2－微球蛋白（β2-MG）、视黄醇结合蛋白（RBP）检测及尿液微量白蛋白（ALB）、N-乙酰基-β—D-氨基葡萄糖苷酶（NAG）、β2-MG、RBP含量检测,并与对照组进行比较。结果观察组血清β2-MG、RBP及尿ALB、NAG、β2-MG含量均高于对照组,尿RBP含量低于对照组（P均〈0．05）。结论SIRS早期即出现肾小球及肾小管的损伤,测定上述指标对于早期监测SIRS患儿肾损伤、及早采取干预措施、阻止病情进展具有重要意义。     

上海中心城区2型糖尿病患者高敏C反应蛋白与白蛋白尿相关性研究

目的 分析上海中心城区2型糖尿病患者高敏C反应蛋白（hs-CRP）与白蛋白尿的相关性.方法 2004年2月至8月,整群抽样上海市中心城区30岁以上已诊断2型糖尿病患者1039例,其中最后人组的574例患者采用免疫比浊法测定了hs-CRP,排除hs-CRP＞10 mg/L者54例,并根据资料完整性,最终有515例患者进行本次分析.采用单因素相关分析和Logistic回归分析评价hs-CRP与白蛋白尿的相关情况.结果 （1）log hs-CRP与年龄、体重指数、腰围、收缩压、舒张压、胆固醇、甘油三酯、白蛋白尿具有相关性（r值分别为0.142、0.305、0.243、0.225、0.264、0.126、0.105、0.168,均P＜0.05）;（2）随着hs-CRP的升高,正常白蛋白尿的比例下降,微量白蛋白尿和大量白蛋白尿的比例升高（x2=18.31,P＜0.01）;（3）组间比较显示,微量白蛋白尿组及大量白蛋白尿组hs-CRP水平与正常白蛋白尿组比较差异有统计学意义（x2值分别为12.36、9.61,均P＜0.01）,微量白蛋白尿组与大量白蛋白尿组相比较,差异无统计学意义（x2=1.24,P＞0.05）.（4）Logistic回归分析显示,收缩压（β值0.035）、hs-CRP（0.110）、HbA1c（0.246）为微量白蛋白尿的独立危险因素（均P＜0.05）.结论 hs-CRP与微量白蛋白尿独立相关,提示慢性亚临床炎症可能参与了社区糖尿病人群早期肾脏损伤.     

甘油三酯和高密度脂蛋白比值与糖尿病肾病的相关性

目的对2型糖尿病患者甘油三酯和高密度蛋白胆固醇的比值与微量白蛋白尿及尿α_1微球蛋白(α_1-MG)的关系进行探讨。方法以2015年3月—2017年11月在该院内分泌科治疗的328例2型糖尿病患者为研究对象,通过查找患者病例采集患者性别、年龄、病况等基本资料以及甘油三酯、高密度蛋白胆固醇、微量蛋白尿、尿α_1微球蛋白等临床资料。尿白蛋白/肌酐比(ACR)3 mg/mmol为非蛋白尿组,ACR≥3 mg/mmo L为蛋白尿组;尿α_1-MG12 mg/L为正常组,尿α_1-MG≥12 mg/L为尿α_1-MG组。采用SPSS 22.0统计学软件进行数据处理,用Logistic回归分析来确定TG/HDL-C与ACR及尿α_1-MG之间的关系。结果与非蛋白尿组相比,在蛋白尿组中SBP、DBP、FPG水平和尿α_1-MG阳性率更高。蛋白尿组的TG和TC以及TG/HDL-C水平很显然比非蛋白尿组的高。与非蛋白尿组相比,在蛋白尿组中SBP、DBP、FPG水平和尿α_1-MG阳性率更高。只有当TG/HDL-C≥2.53时,差异有统计学意义(P0.05)。结论与低水平相比,高水平的TG/HDL-C与ACR、尿α_1-MG均存在正相关关系。     

High concentrations of human β-defensin 2 in gastric juice of patients with Helicobacter pylori infection

AIM: Human β-defensin (HBD)-1 and HBD-2 are endogenous antimicrobial peptides. Unlike HBD-1, the HBD-2 expression is augmented by Helicobacter pylori (H pylori). We sought to determine HBD-1 and HBD-2 concentrations in gastric juice duringH pylori infection.METHODS: HBD-1 and HBD-2 concentrations were measured by radioimmunoassay in plasma and gastric juice of 49 H pylori-infected and 33 uninfected subjects and before and after anti-H pyloritreatment in 13 patients with H pylori-associated gastritis. Interleukin (IL)-1β and IL-8 concentrations in gastric juice were measured by enzyme-linked immunosorbent assay (ELISA). Histological grades of gastritis were determined using two biopsy specimens taken from the antrum and corpus. Reverse phase high performance liquid chromatography (RP-HPLC)was used to identify HBD-2.RESULTS: HBD-2 concentrations in gastric juice, but not in plasma, were significantly higher in H pylori-positive than -negative subjects, albeit the post-treatment levels were unchanged. Immunoreactivity for HBD-2 was exclusively identified in H pylori-infected mucosa by RPHPLC. HBD-2 concentrations in gastric juice correlated with histological degree of neutrophil and mononuclear cell infiltration in the corpus. IL-1β levels correlated with those of IL-8, but not HBD-2. Plasma and gastric juice HBD-1concentrations were similar in H pylori-infected and uninfected subjects.CONCLUSION: Our results place the β-defensins, especially HBD-2, in the front line of innate immune defence.Moreover, HBD-2 may be involved in the pathogenesis of H pylori-associated gastritis, possibly through its function as immune and inflammatory mediator.     

Clinical,radiological, and biochemical features of a bilateral buttock amyloidoma emerging after 27 years of hemodialysis

Deposition of amyloid in the buttock is a rare complication of dialysis related amyloidosis (DRA), but this localization is even rarer in other types of amyloidoses. We report here the clinical, radiological, and biochemical features of a patient who incurred into this complication after 27 years of hemodialysis. Imaging of the amyloid deposition by magnetic resonance imaging (MRI) documents the amyloid infiltration in the muscles of the buttock region and highlights a peculiar feature of amyloid fibrils deposition in the subcutaneous fat. The amyloid deposition is confirmed by biochemical and microscopic analysis of fibrils extracted from a biopsy specimen. Review of literature and the features of this case lead to speculation that the peculiar involvement of the buttock region including muscles and subcutaneous fat in DRA might derive from the propagation of amyloid initially deposited in the hip joint.     

β-防御素-2与Toll样受体关系的研究进展

β-防御素-2是固有免疫应答的重要组成成分,是-类可诱导性的抗菌肽,抗菌谱广,对革兰阳性菌、革兰阴性菌、真菌、分枝杆菌、非典型病原体和包膜病毒都具有-定的抵抗作用。关于β防御素-2的生成机制虽然尚未完全阐明,但大量研究表明与上皮细胞表达的Toll样受体密切相关,本文就β防御素-2与Toll样受体关系的研究进展作一综述。     

原发性肝癌患者血清β2-微球蛋白含量变化的临床意义

目的探讨原发性肝癌患者血清β2-微球蛋白（β2-microglobulin,β2-MG）含量变化及临床应用价值。方法用免疫放射分析法（RIA）检测64例原发性肝癌患者、55例肝硬化、以及48例正常健康人血清β2-MG含量;动态检测原发性肝癌手术前后β2-MG含量变化。结果原发性肝癌组血清中β2-MG含量明显高于健康对照组（P〈0.05）和肝硬化组（P〈0.05）;原发性肝癌术前血清β2-MG含量增高者,术后含量明显降低,有转移复发时,β2-MG含量再度升高。结论检测血清β2-MG含量变化在原发性肝癌辅助诊断以及疗效、复发判断方面具有重要临床意义。     

卡维地洛对急性心肌梗死大鼠心肌组织中β-arrestin 2蛋白表达的影响

目的:探讨卡维地洛对急性心肌梗死(MI)大鼠非梗死区心肌组织中β-arrestin 2的影响。方法:30只大鼠随机分成假手术组、模型组和卡维地洛组。建立MI模型,术前3d至术后4d给予药物及0.9%氯化钠,实验结束后测量心功能和非梗死区心肌组织中β-arrestin 2的表达。结果:与假手术组相比,模型组大鼠左室收缩末期压(110.72±13.27)mmHg(1mmHg=0.133kPa)、左室压力最大上升速率(2.62±0.83)和下降速率(-2.11±0.57)明显降低(P<0.05)。与模型组比较,卡维地洛对心功能有明显改善作用,可使左室收缩末期压(130.78±7.94)mmHg明显升高(P<0.05)、左室舒张末期压[(12.34±5.55)mmHg]明显降低(P<0.05),并升高了左室压力最大上升速率和下降速率(P<0.05)。模型组大鼠非梗死区心肌β-arrestin 2蛋白表达增加(P<0.05),卡维地洛可进一步增加非梗死区心肌β-arrestin 2蛋白表达(P<0.05)。结论:卡维地洛可明显改善大鼠MI后左室功能,其机制可能与其增加MI后非梗死区心肌组织中β-arrestin 2的表达有关。     

Relationship and significance between anti-β2-glycoprotein Ⅰ antibodies and platelet activation state in patients with ulcerative colitis

AIM： To study the relationship between anti-β2- glycoprotein Ⅰ （aβ2GPⅠ） antibodies and platelet activation state in patients with ulcerative colitis （UC） and its significance.
METHODS： Peripheral blood samples were collected from 56 UC patients （34 males and 22 females, aged 43.5 years, range 21-66 years）, including 36 at active stage and 20 at remission stage, and 25 sex-and age-matched controls. The level of aβ2GP Ⅰ was measured by ELISA. The platelet activation markers, platelet activation complex- Ⅰ （PAC- Ⅰ ） and P-selectin （CD62P） were detected by flow cytometry.
RESULTS： The A value for IgG aβ2GP Ⅰ in the active UC group was 0.61 ± 0.13, significantly higher than that in the remittent UC and control groups （0.50 ± 0.13 and 0.22 ± 0.14, P 〈 0.01）. There was a significant difference between the two groups （P 〈 0.01）. The A value for IgM aβ2GP Ⅰ in the active and remittent UC groups was 0.43 ± 0.13 and 0.38 ± 0.12, significantly higher than that in the control group （0.20 ± 0.12, P 〈 0.01）. However, there was no significant difference between the two groups （P 〉 0.05）. The PAC- Ⅰ positive rate for the active and remittent UC groups was 30.6% ± 7.6% and 19.6% ± 7.8% respectively, significantly higher than that for the control group （6.3% ± 1.7%,P 〈 0.01）. There was a significant difference between the two groups （P 〈 0.01）. The CD62P positive rate for the active and remittent UC groups was 45.0% ± 8.8% and 31.9% ± 7.8% respectively, significantly higher than that for the control group （9.2% ± 2.7%, P 〈 0.01）. There was a significant difference between the two groups （P 〈 0.01）. In the active UC group, the more severe the state of illness was, the higher the A value for IgG aβ2GP Ⅰ was, and the positive rate for PAC-Ⅰ and CD62P was positively correlated with the state of illness （Faβ2GP Ⅰ = 3.679, P 〈 0.05; FPAC-Ⅰ （%） = 5.346, P 〈 0.01; and FCD62P （%） = 5. 418, P 〈 0.01）. Mea     

First Identification of a Point Mutation at Position −83 (G>A) of the β-Globin Gene Promoter

We report the first identification of a point mutation located within the promoter region of the β‐globin gene at position ?83 (G>A) and associated with the common heterozygous deletional α‐thalassemia (α-thal) (?α^3.7/αα). The patient was an adult male from Gabon belonging to the Obamba sub ethnic group, who was referred to our clinics for a mild microcytic anemia with a Hb A₂ level at the upper limit of the normal value (3.5%). This observation is a new example of α- and β-thal co-inheritance with a normal Hb A₂ level, and illustrates a potential source of pitfall in screening for α- and β-thal carriers.     

Genetic modulation of islet β-cell iPLA2β expression provides evidence for its impact on β-cell apoptosis and autophagy

β-cell apoptosis is a significant contributor to β-cell dysfunction in diabetes and ER stress is among the factors that contributes to β-cell death. We previously identified that the Ca²⁺-independent phospholipase A₂β (iPLA₂β), which in islets is localized in β-cells, participates in ER stress-induced β-cell apoptosis. Here, direct assessment of iPLA₂β role was made using β-cell-specific iPLA₂β overexpressing (RIP-iPLA₂β-Tg) and globally iPLA₂β-deficient (iPLA₂β-KO) mice. Islets from Tg, but not KO, express higher islet iPLA₂β and neutral sphingomyelinase, decrease in sphingomyelins, and increase in ceramides, relative to WT group. ER stress induces iPLA₂β, ER stress factors, loss of mitochondrial membrane potential (?Ψ), caspase-3 activation, and β-cell apoptosis in the WT and these are all amplified in the Tg group. Surprisingly, β-cells apoptosis while reduced in the KO is higher than in the WT group. This, however, was not accompanied by greater caspase-3 activation but with larger loss of ?Ψ, suggesting that iPLA₂β deficiency impacts mitochondrial membrane integrity and causes apoptosis by a caspase-independent manner. Further, autophagy, as reflected by LC3-II accumulation, is increased in Tg and decreased in KO, relative to WT. Our findings suggest that (1) iPLA₂β impacts upstream (UPR) and downstream (ceramide generation and mitochondrial) pathways in β-cells and (2) both over- or under-expression of iPLA₂β is deleterious to the β-cells. Further, we present for the first time evidence for potential regulation of autophagy by iPLA₂β in islet β-cells. These findings support the hypothesis that iPLA₂β induction under stress, as in diabetes, is a key component to amplifying β-cell death processes.     

Formation of Cross-Linked Asymmetrical Hybrid Hemoglobins by Double-Headed Aspirin

Double-headed aspirin [bis(3,5-dibromosalicyl) fumarate] selectively cross-links hemoglobin molecules between Lys 82β₁ and Lys 82β₂ and increases solubility of deoxy-Hb S (Walder et al., J. Mol. Biol., 141:195, 1980 and Kikugawa et al., J. Biol. Chem., 257: 7525, 1982). We reacted this reagent with the mixture of Hb A and Hb S and the mixture of Hb S and Hb York (β146His→Pro). Crosslinked asymmetrical hybrid hemoglobins (α₂ β-β ^S and α₂ β ^Y-β ^S) were produced in high yields in addition to the cross-linked parent hemoglobin molecules. Results on electrophoresis, gel electrofocusing, ion exchange column chromatography, mechanical stability and oxygen binding properties showed that the cross-linked asymmetrical hybrid hemoglobins had properties intermediate between those of the cross-linked parent hemoglobins. Oxygen affinities of the cross-linked asymmetrical hybrids were not affected by the addition of 2,3-diphosphoglycerate (DPG) or inositol hexaphosphate, probably due to the presence of a fumaryl group at the DPG binding site.