心力衰竭患者自然杀伤细胞活性的变化

目的探讨充血性心力衰竭（CI-IF）患者自然杀伤（NK）细胞活性的变化。方法选择NYHAⅡ-Ⅳ级的CI-IF患者48例作为观察组，30例健康查体者作为对照组。应用流式细胞仪测定全血NK细胞数目，应用改良MIT法测定NK细胞杀伤活性。结果CHF患者循环NK细胞和NK细胞杀伤活性显著降低，并与心力衰竭程度呈负相关（r=-0．873、-0．949，P均＜0．001）。结论CHF患者NK细胞数目和NK细胞活性显著降低，可能是CHF患者免疫功能异常的机制之一。     

充血性心力衰竭患者自然杀伤细胞活性的变化

目的探讨充血性心力衰竭（CHF）患者自然杀伤（NK）细胞活性的变化。方法选择纽约心脏病学会（NYHA）Ⅱ～Ⅳ级的CHF患者48例作为观察组，30例健康体检者作为对照组。采集清晨空腹静脉血，应用流式细胞仪测定全血NK细胞数目，应用改良四甲基偶氮唑盐（MTF）比色法测定NK细胞杀伤活性。结果CHF患者循环NK细胞和NK细胞杀伤活性显著降低（P〈0．01或P〈0．001），并与心力衰竭程度呈负相关（r=-0．873、-0．949，P均〈0．001）。结论CHF患者NK细胞数目和NK细胞活性显著降低，可能是CHF患者免疫功能低下的原因。     

心力衰竭患者NK细胞活性变化

目的 探讨慢性心力衰竭（CHF）患者自然杀伤细胞（NK）活性的变化。方法 选择NYHAⅡ-Ⅳ级的CHF患者48例作为观察组,30例健康查体者作为对照组。测定全血NK细胞数目和血清、培养上清中白细胞结素6（IL-6）含量,并测定NK细胞杀伤活性。结果 （1）CHF患者循环NK细胞和NK细胞杀伤活性显著降低,并与心衰程度呈负相关（r=-0.873,-0.949,均P〈0.001）;（2）CHF患者血清、培养上清和IL-2刺激后上清中IL-6显著增加（均P〈0.001）,并与心衰程度呈正相关（r=0.988,0.984,0.982;均P〈0.001）;（3）IL-2刺激可能使CHF患者NK细胞杀伤活性增强（P〈0.001）。结论 NK细胞数目和活性下降可能是CHF患者免疫功能异常的机制之一。     

重组生长激素对烧伤患者NK细胞数量、活性和NK细胞毒因子活性影响的研究

目的探讨重组生长激素（rhGH）改善烧伤患者的免疫状况的作用。方法42例严重烧伤患者随机分成2组，实验对照组22例采用常规治疗，治疗组20例在常规治疗基础上应用rhGH治疗；并分别采用碱性磷酸酶抗碱性磷酸酶（APAAP）免疫细胞化学法、乳酸脱氢酶（LDH）释放法和四甲基偶氮唑盐（MTT）比色法检测了2组烧伤患者伤后24h内和用药后第7、14天外周血的自然杀伤（NK）细胞数量、NK细胞活性和NK细胞毒因子（NKCF）活性。另取20名体检合格的健康献血员作为正常对照组。结果烧伤后24h内，2组患者上述3项指标均明显低于正常对照组（P〈0．01）。用药后第7、14天，实验对照组上述3项指标仍处在很低水平，而治疗组上述3项指标已基本恢复正常，两组比较，差异有显著性（P〈0．01）。结论rhGH可明显提高烧伤患者外周血的NK细胞数量、NK细胞活性和NKCF活性，能够提高烧伤患者受抑制的免疫功能。     

NK细胞活性在诊断继发性噬血细胞综合征中的意义探讨

本研究探讨继发性噬血细胞综合征（hemophagocytic syndrome，HPS）患者外周血NK细胞活性水平及其在早期诊断中的意义。采用LDH释放法检测16例凝似HPS患者和25名正常健康人外周血NK细胞的活性，并将确诊继发性HPS患者的NK细胞活性与正常人NK细胞活性进行比较。结果表明：16例疑似HPS患者中有8例最终确诊为继发性HPS，其NK细胞活性均明显低于正常对照组，两者相比差异有显著统计学意义（p〈0．001），且已确诊患者的NK细胞活性在疾病早期即出现异常。结论：NK细胞活性的检测对于HPS的早期诊断可能具有重要意义。     

免疫球蛋白样抑制性受体KIR基因与HLA-Cw的匹配对NK细胞活性的影响

本研究探索自然杀伤（naturalkiller,rqK）细胞表面杀伤细胞免疫球蛋白样抑制性受体（killerimmunoglobulin—like inhibitionreceptor,KIR）基因与HLA—Cw配体匹配对NK细胞活性的影响。对27名健康人取新鲜外周血20ml,用NK细胞免疫磁珠分选试剂盒负向分选高纯度NK细胞。对30例初治确诊急性髓系白血病（AML）患者取新鲜骨髓液10ml,分离单个核细胞作为靶细胞。流式细胞仪检测NK细胞CD158a,CD158b表达水平。取健康人和患者的外周血各2ml,以PROTRANS法抽提DNA,采用序列特异性引物PCR—SSP分型技术分别检测HLA—Cw、KIR基因。MTT法检测KIR与HLA—Cw基因不同组合的NK细胞对AML患者白血病细胞的杀伤率。结果表明：分选后的NK细胞,纯度为（90．8±6．08）％。NK细胞KIR基因与靶细胞HLA—Cw等位基因的匹配数少则NK细胞的杀伤效应强,0个等位基因匹配的杀伤率为（50．66±8．40）％,1个匹配与2个匹配的杀伤率分别为（38．28±6．71）％,（19．74±4．15）％,（F=20．226,P〈0．001）。NK细胞的KIR表达水平与杀伤作用也有一定的联系（F=16．276,P值〈0．001）。结论：NK细胞对AML白血病细胞的杀伤作用与抑制性KIR／HLA—Cw的匹配数及KIR的表达相关,匹配越少、KIR表达越低,杀伤率越高。     

高能聚焦超声对恶性肿瘤细胞免疫功能的影响

目的：通过检测NK细胞活性及T淋巴细胞亚群的变化，探讨高能聚焦超声（High intensity focused ultrasound,HIFU)治疗对人体细胞免疫功能的影响。方法：利用流式细胞仪检测23例恶性肿瘤患者HIFU治疗前后NK细胞活生及T淋巴细胞亚群的变化，结果：治疗后NK细胞活性显著增强（P＜0．05），CD3^ ,CD34^ ，CD4^ ／CD8^ 有不同程度的增加，同治疗前相比无显著差异（P＞0．05）。其中14例的发性恶性肿瘤且无明显远处转移的患者单纯行HIFU治疗后CD4^ ，CD4^ ／CD8^ 显著增多。HIFU同时联合应用化疗者经HIFU治疗后CD3^ 、CD4^ 轻度降低，CD4^ ／CD8^ 略有升高。结论：HIFU治疗可以显著增强NK细胞的活性，增加T淋巴细胞亚群数量，增强机体细胞免疫功能。     

放化疗对食管癌患者T淋巴细胞及自然杀伤细胞影响的临床研究

目的探讨食管癌患者外周血T淋巴细胞、自然杀伤（NK）细胞的变化及放化疔对其的影响。方法68例经病理组织学确诊的食管癌患者采用单纯放疗或同步放化疗治疗,应用流式细胞仪测定其治疗前后外周血T淋巴细胞亚群及NK细胞比例,以健康体检人群为对照分析其变化。结果（1）食管癌患者外周血T细胞、Th细胞、Th／Ts细胞比值较正常对照组均明显下降（P〈0．01）,而Ts细胞水平显著升高（P〈0．05）。（2）外周血T淋巴细胞亚群及Th／Ts比例与患者性别、肿瘤分期、病理分化程度、病变部位等临床病理参数无明显相关性。（3）放化疗后,患者外周血总T细胞、Th细胞、Th／Ts细胞比值及NK细胞均上升（P〈0．01）,而Ts细胞降低（P〈0．05）;单放组总T细胞、Th细胞、NK细胞较同步组升高更显著。结论食管癌患者细胞免疫功能低下,检测T淋巴细胞亚群、NK细胞可用于患者的免疫监测。放疗及化疗应着重围绕提高总T细胞、Th细胞和NK细胞的数量与功能。     

卵巢恶性肿瘤患者外周血T淋巴细胞亚群NK细胞活性检测的临床意义

目的 探讨卵巢恶性肿瘤的外周血T细胞亚群和NK细胞活性的变化。方法 利用流式细胞仪测定52例卵巢恶性肿瘤患者和50例妇科良性肿瘤患者的外周血T细胞亚群和NK细胞活性对照比较。结果 卵巢恶性肿瘤患者CD3、CD4和NK细胞水平较妇科良性肿瘤患者组显著下降（P＜0．05），CD4／CD8比值明显低于对照组（P＜0．01）。卵巢恶性肿瘤组CD3水平虽随着临床分期增加而下降，但无统计学意义（P＞0．05），而CD4、CD4／CD8和NK细胞则随着临床分期增加而明显下降（P＜0．05），CD8水平随着临床分期增加而明显上升（P＜0．05）。结论 卵巢恶性肿瘤患者细胞免疫功能较妇科良性肿瘤患者低，并且细胞免疫功能与肿瘤的负荷有密切关系，随着临床分期增加而明显下降。     

合并腹主动脉旁淋巴结清扫的胃癌扩大根治术与D2手术前后细胞免疫功能的变化

目的 探讨进展期胃癌（AGC）患者行合并腹主动脉旁淋巴结（PAL）清扫的胃癌扩大根治术（D3＋PAL）前后细胞免疫功能的变化，并与D2手术后的细胞免疫功能进行对比分析。方法 应用改良M1Tr法和T淋巴细胞亚群SAP法检测104例AGC患者和33例正常对照（对照组）外周血的NK细胞杀伤活性及T淋巴细胞亚群的变化。结果 与对照组相比，2组AGC患者手术前NK细胞活性，CD3^＋、CD4^＋．CD4^＋／CD8^＋明显降低（CD8^＋升高），术后D3＋PAL组NK细胞活性，CD3^＋、CD4^＋、CD4^＋／CD8^＋比术前明显升高（P〈0．05或0.01）；与D2术后相比．D3＋PAL术后NK细胞活性，CD3^＋、CD4^＋、CD4^＋／CD8^＋明显升高。结论 手术前AGC患者细胞免疫功能明显降低，D3＋PAL手术可明显改善AGC患者的细胞免疫功能，且比D2手术效果明显。     

造血与淋巴组织肿瘤患者红细胞的天然免疫黏附功能及对NK细胞杀伤活性的影响

本研究探讨造血与淋巴组织肿瘤患者红细胞天然免疫黏附功能及红细胞对NK细胞杀伤活性的影响。外周血枸橼酸钠抗凝,检测红细胞在自身血浆条件下对K562细胞的免疫黏附并计算黏附率。用MTT法测定红细胞对正常NK细胞杀伤K562细胞活性的影响,并与未加红细胞时进行比较。结果表明:红细胞对K562细胞形成了"玫瑰花"样结合。正常人红细胞对K562细胞的免疫黏附结合率为(15.3±6.4)%,造血与淋巴组织肿瘤患者红细胞对K562细胞的免疫黏附结合率为(7.6±7.0)%,与正常人相比较显著下降(t=3.61,p(0.001)。不加红细胞条件下,正常人外周静脉血NK细胞杀伤K562细胞活性为67%-71%。正常人红细胞明显增加NK细胞杀伤K562细胞活性杀伤率为(14.7±5.2)%,造血与淋巴组织肿瘤患者的红细胞减低NK细胞杀伤K562细胞活性,杀伤率为(4.3±7.6)%,二者比较有显著差异(t=6.73,p(0.0001)。结论:造血与淋巴组织肿瘤患者红细胞对K562细胞的免疫黏附能力下降,正常人红细胞明显增加NK细胞杀伤K562细胞活性,而造血与淋巴组织肿瘤患者的红细胞减低NK细胞杀伤K562细胞活性,原因需进一步研究。     

风湿性心脏病NK细胞活性的变化及其临床意义

本文对50例风湿性心脏病患者外周血NK细胞活性的变化进行了研究,结果:风湿活动组NK细胞活性明显降低,而非风湿活动组NK活性与对照组相比无显著差异.认为NK细胞活性降低与免疫反应调节失控有关,同时NK细胞功能缺陷本身,又进一步加剧了免疫调节功能紊乱,加重了心肌及心瓣膜的免疫损害.     

慢性心力衰竭患者体质量指数改变的临床意义

目的 观察慢性心力衰竭患者治疗前后体质量指数(body mass index, BMI)改变情况,明确其改变的临床指导意义。 方法 共纳入65例慢性心力衰竭患者,患者入院次日晨空腹测量其体质量、身高,计算BMI,其后每日测量体质量,直至病情好转出院。计算患者入院后BMI最高值与最低值的差值、观察并记录患者入院时的临床表现,如心功能分级、水肿程度。 结果 患者在出院时BMI值显著下降(t=9.083,P<0.001)。 心功能Ⅳ级患者,BMI差值显著高于Ⅱ、Ⅲ级患者(Z=3.655,P<0.001; Z=4.562,P<0.001);重度水肿患者BMI差值高于轻、中度水肿患者,同时三者均高于无水肿患者。结论 BMI的变化可作为衡量心力衰竭严重程度的指标,有临床应用价值。     

Cardiac secretion of adrenomedullin in human heart failure.

Adrenomedullin (ADM) is a newly discovered endogenous vasorelaxing and natriuretic peptide. Recently, we have reported that plasma ADM is increased in severe congestive heart failure (CHF) in humans and that increased immunohistochemical staining is observed in the failing human ventricular myocardium. The present study was designed to test the hypothesis that the failing human ventricle secretes ADM and that circulating ADM progressively increases with the severity of clinical CHF. Plasma ADM was significantly increased in human CHF (39.8 +/- 3.6 pg/ml, P < 0.001 vs. normal) as compared with normal subjects (14.4 +/- 2.7 pg/ml). Plasma ADM was increased in mild CHF (NYHA class II, 30.1 +/- 3.4 pg/ml, P < 0.01 vs. normal), moderate CHF (NYHA class III, 31.5 +/- 3.0 pg/ml, P < 0.01 vs. normal), and severe CHF (NYHA class IV, 66.1 +/- 9.4 pg/ml, P < 0.001 vs. normal). In 13 patients with CHF in whom plasma samples were obtained from aorta (AO), coronary sinus (CS) and anterior interventricular vein (AIV), there was a significant step-up in plasma ADM between AO and AIV (50.6 +/- 9.3 pg/ml and 62.1 +/- 11.1 pg/ml, respectively, P < 0.01) and between AO and CS (50.6 +/- 9.3 pg/ml and 58.6 +/- 11.4 pg/ml, respectively, P < 0.05). The current study demonstrates that the failing human heart secretes ADM in human CHF suggesting contribution to the increase in plasma ADM, and indicates for the first time an additional endocrine system of cardiac origin which is activated in human CHF and may function in cardiorenal regulation.     

氨基末端脑钠肽前体在心力衰竭检测中的临床意义

目的:探讨充血性心力衰竭患者血浆氨基末端脑钠肽前体(NT-proBNP)水平的变化。方法:应用电化学发光免疫分析仪测定健康人组80例和心力衰竭组80例(心功能Ⅱ级25例、Ⅲ级30例、Ⅳ级25例)血清NT-proBNP的水平。结果:心力衰竭组NT-proBNP水平较健康组明显增高,且随心功能的恶化NT-proBNP水平增高。结论:充血性心力衰竭患者血清NT-proBNP水平明显增加,且随着心功能分级程度的加重而显著增加。     

Measuring baroreflex sensitivity from the gain function between arterial pressure and heart period

We tested an innovative approach for estimating baroreflex sensitivity (BRS) from the gain function between spontaneous oscillations of systolic arterial pressure (SAP) and heart period (HP). The major goal was to assess the practical implications of abandoning the classical coherence criterion (> or =0.5) as regards measurability of BRS, and agreement with values of BRS obtained using the phenylephrine test (Phe-BRS). We studied 19 normal subjects, 44 patients with a history of previous myocardial infarction (MI) and 45 patients with chronic heart failure (CHF). The experimental protocol included recording of SAP and HP for 10 min of supine rest, and evaluation of Phe-BRS. From resting SAP and HP, the gain and coherence functions were computed. The new BRS index was obtained in all subjects by averaging the gain function over the whole low-frequency band (0.04-0.15 Hz) (whole-band average BRS, WBA-BRS). WBA-BRS was 7.4 (5.8-10.8) ms/mmHg [median (25th-75th percentile)] in normal controls, 3.1 (1.4-5.4) ms/mmHg in MI patients (P<0.001 compared with normals) and 5.0 (3.2-6.9) ms/mmHg in CHF patients (P<0.01 compared with normals). Using the coherence criterion, BRS could be measured in only 43% and 49% of MI and CHF patients respectively, and the proportion of the low-frequency band contributing to the measurement was 21% (14-47%) and 29% (16-35%) respectively. The correlation between WBA-BRS and Phe-BRS was 0.47, 0.63 and 0.36 in the normal, MI and CHF groups respectively (all P<0.001). The relative bias of WBA-BRS was -5.2 ms/mmHg (P<0.001) in normals, -1.4 ms/mmHg (P=0.004) in MI patients and -1.0 ms/mmHg (P=0.11) in CHF patients. The limits of agreement were -13 to 2.6, -7.4 to 4.6 and -9.3 to 7.3 ms/mmHg in the normal, MI and CHF groups respectively. Thus the WBA-BRS method standardizes the computation of BRS among subjects, and dramatically increases its measurability in subjects with pathology compared with the classical spectral technique based on the coherence criterion. Compared with Phe-BRS, WBA-BRS tends to give negatively biased results. The correlation and the magnitude of the limits of agreement between the two methods are similar to those observed previously using coherence-based spectral methods.     

Rescue of Impaired NK Cell Activity in Hodgkin Lymphoma With Bispecific Antibodies In Vitro and in Patients

Natural killer (NK) cells represent a key component of the innate immune system against cancer. Nevertheless, malignant diseases arise in immunocompetent individuals despite tumor immunosurveillance. Hodgkin lymphoma (HL) is characterized by CD30⁺ tumor cells and a massive infiltration of immune effector cells in affected lymph nodes. The latter obviously fail to eliminate the malignant cell population. Here, we tested for functional NK cell defects in HL and suggest an improvement of NK function by therapeutic means. We demonstrate that peripheral NK cells (pNK) from patients with HL fail to eliminate HL cell lines in ex vivo killing assays. Impaired NK cell function correlated with elevated serum levels of soluble ligands for NK cell receptors NKp30 (BAG6/BAT3) and NKG2D (MICA), factors known to constrict NK cell function. In vitro, NK cell cytotoxicity could be restored by an NKG2D/NKp30-independent bispecific antibody construct (CD30xCD16A). It artificially links the tumor receptor CD30 with the cytotoxicity NK cell receptor CD16A. Moreover, we observed that NK cells from patients treated with this construct were generally activated and displayed a restored cytotoxicity against HL target cells. These data suggest that reversible suppression of NK cell activity contributes to immune evasion in HL and can be antagonized therapeutically.     

血浆BNP检测对心力衰竭患者心功能分级评价中的作用

目的探讨血浆B型钠尿肽(B-type natriuretic peptide,BNP)浓度在诊断慢性心力衰竭(chronic heart fail-ure,CHF)及心功能分级评价中的作用。方法采用化学发光微粒子酶免疫分析法测定89例CHF患者及40名健康对照者血浆BNP浓度。依据美国纽约心脏病协会(New York Heart Association,NYHA)分级方案对CHF患者进行心功能分级。结果血浆BNP诊断CHF临界值为111.7pg/ml,灵敏度88.16%,特异性94.29%;CHF组血浆BNP浓度为574.3±176.2pg/ml,明显高于对照组66.7±17.5pg/ml,P<0.05。CHF患者NYHA心功能Ⅰ级到Ⅳ级血浆BNP逐渐升高,各级之间BNP浓度比较,P<0.05;血浆BNP水平与NYHA分级间存在明显的正相关,r=0.835,P<0.001。各级NYHA分级CHF患者经治疗后血浆BNP水平显著下降,与治疗前相比,P<0.05。结论 CHF患者血浆BNP水平随着心力衰竭严重程度的增加而升高,血浆BNP可以准确评价心功能,可作为临床诊断、疗效观察以及预后判断CHF的客观指标。