不同抗凝及保存方式的全血测定HbA1c结果分析

糖化血红蛋白A1c（HbAlc）是评价糖尿病长期控制情况的良好指标。文献曾报道正常人全血用EDTA-K2、枸橼酸三钠抗凝的结果比对[1],对于糖尿病患者样本的抗凝剂选择的则没有。3.2%枸橼酸钠1∶4（0.4 mL枸橼酸钠＋1.6 mL全血）、3.2%枸橼酸钠1∶9（0.3 mL枸橼酸钠＋2．7mL全血）都是临床上常见的抗凝剂。     

不同封管方法在血液透析留置导管并高危出血患者中的应用

目的:观察不同浓度肝素和枸橼酸封管用于血液透析留置导管合并高危出血患者的效果比较。方法:选取血液透析合并高危出血患者90例,并随机分为三组,Ⅰ组:肝素浓度3125μ/mL组;Ⅱ组:肝素浓度1250μ/mL组;Ⅲ组:4%枸橼酸钠组。观察插管前及封管后1h、4h凝血时间(PT、APTT、INR),比较三组间各时间段的凝血时间差异,比较三组间导管出血、导管堵塞、导管感染情况。结果:Ⅰ组高浓度肝素组在封管后1h的PT、APTT、INR较基础值显著延长(P<0.01),封管后4h的凝血指标较1h有所下降,仍显著高于基础值(P<0.05);Ⅱ组低浓度肝素组在封管后1h的PT、APTT显著超过基础值(P<0.05),而4h后的凝血指标与基础值比较差异无统计学意义(P>0.05);Ⅲ组枸橼酸组封管后1h、4h的PT、APTT、INR与基础值比较差异均无统计学意义(P>0.05);Ⅰ、Ⅱ组与Ⅲ组在出血方面比较,差异有统计学意义(P<0.05),而导管堵塞、感染方面,三组比较差异无统计学意义(P>0.05)。结论:血液透析留置导管合并高危出血倾向患者,采用4%枸橼酸封管,可明显降低出血风险,导管并发症少。     

RNAⅢ抑制肽/磷酸胆碱基细胞膜仿生药物涂层的血液相容性

背景:课题组在国内首次合成了葡萄球菌抑制剂RNAⅢ抑制肽,并制备了RNAⅢ抑制肽,磷酸胆碱基细胞膜仿生药物涂层.目的:评价RNAIII抑制肽,磷酸胆碱基细胞膜仿生药物涂层的血液相容性.设计、时间及地点:随机对照动物实验,于2005-10/2007-10在解放军总医院临床药理研究所及医学动物实验中心完成.材料:成年普通级健康新西兰大白兔30只,由解放军总医院动物实验中心提供.方法:将洁净的316L不锈钢片浸渍于5 g/L的四元共聚物四氢呋喃和质量分数10%的RNAⅢ抑制肽混合溶液中,获得稳定的聚合物涂层,按1 cm~2/mL在37℃无菌条件下用生理盐水浸提72 h,制得浸提液原液;加入同体积的无菌生理盐水制得50%浸提液.选择溶血实验、凝血实验、血小板聚集实验,测定凝血酶原时间和活化部分凝血酶时间、观察RNAIII抑制肽/磷酸胆碱基细胞膜仿生药物涂层对兔白细胞、红细胞和血小板的影响.主要观察指标:红细胞溶血率,凝血时间、凝血酶原时间、活化部分凝血酶时间,白细胞、红细胞和血小板数量,血小板聚集情况.结果:溶血实验结果显示浸提液原液和50%浓度浸提液的溶血率分别为3.08%和1.88%,两者的溶血率均<5%,符合医用生物材料的溶血实验要求.凝血实验结果表明浸提液原液和50%浓度浸提液对兔凝血时间无明显影响,对各时间点兔凝血酶原时间和活化部分凝血酶时间均无明显作用,对兔白细胞、红细胞和血小板数量无明显影响,对兔血小板聚集无明显影响.结论:实验数据表明,国内首次合成的RNAⅢ抑制肽,磷酸胆碱基细胞膜仿生药物涂层具有良好的血液相容性.     

镁合金材料的溶血率及影响因素

背景:镁合金作为潜在的新型医用可降解生物金属材料受到越来越多的关注,体外纯镁合金浸提液具有溶血作用.目的:探讨镁合金浸提液与其溶血率的关系.方法:将镁合金浸提液稀释,按GBT 16175-2008<医用有机硅材料生物学评价实验方法>第13 部分<溶血试验>的改良方法进行溶血实验.测量A值,计算溶血率,各稀释度溶液钠、镁离子浓度及pH值.结果与结论:100%,50%,25%,10%,5%,2.5%和1%稀释度镁浸提液的溶血率分别为65.3%,66.7%,33.8%,11.8%,6.2%,0.3%和-0.3%.随着稀释度的加大,镁离子浓度呈下降趋势,而钠离子则无明显波动.100%,50%,25%,10%,5%和2.5%稀释度之间的pH值差异无显著性意义(P>0.05),均高于1%稀释度(P<0.05).提示镁合金浸提液的高pH值和离子浓度是影响其体外试验溶血的主要因素.     

术后危重患者血滤导管封管液与出血相关并发症的研究

目的 研究4%枸橼酸钠、4 000 U/mL肝素钠、6 250 U/mL肝素钠三种封管液对进行血滤治疗的外科术后危重患者的影响,分析封管液配方与出血相关并发症的关系。方法 纳入北京大学第一医院外科重症病房2014～2016年29个月行床旁血滤患者116例,进行回顾性队列研究。比较使用4%枸橼酸钠、4 000 U/mL肝素钠、6 250 U/mL肝素钠对血滤导管封管的影响。观察的结果包括外科术后3 d和10 d失血指数、术后显性失血量、导管通畅性、封管后凝血功能和导管感染率。结果 在研究期间,共有116名患者接受了439次血滤后封管。使用6 250 U/mL肝素钠封管后3 d和10 d失血指数和显性失血量均明显高于4%枸橼酸组(P0.001)和4 000 U/mL肝素钠组(P0.001)。三组封管液封管后PT均无显著变化,P=0.833。APTT在4%枸橼酸组和6 250 U/mL肝素钠组有显著性差异,P=0.023。4%枸橼酸盐组的导管功能障碍比两个浓度的肝素钠组高6.91%vs4.92%vs2.98%,P0.05,但无统计学意义。三组封管液对于导管相关感染无差异,P0.05。结论 4%枸橼酸钠和4 000 U/mL肝素钠对于外科手术相关出血并发症和全身凝血的影响小于6 250 U/mL肝素钠;三组封管液在维持血滤患者的导管通畅性方面有效性类似。三种封管液在导管相关感染方面安全性相似。     

血塞通注射液溶血性考察与研究

目的对市场上流通使用的血塞通注射液在溶血性方面的状况进行考察与研究。方法按《中国药典》2005年版一部附录ⅩⅧB中药注射剂安全性检查法应用指导原则和中药、天然药物刺激性和溶血性研究的技术指导原则,对11个厂家共计27批血塞通注射液每批次样品制备4个浓度,进行溶血实验研究,并采用分光光度法（545λ/nm）测定计算溶血率,比较各批次样品的溶血率。结果不同厂家甚至同一厂家不同批次的血塞通注射液溶血率存在一定差异。结论在临床使用中应注意用量,过量使用可能导致溶血引起的不良反应;同时,应注意溶血引起的临床不良反应的观测。     

兔抗羊抗体血清的制备及其检测方法的实验方案

目的:制备兔抗羊抗体血清,应用试管凝集实验和试管溶血实验测定其效价,并对其进行了初步探讨。方法:对兔2%～5%SRBC进行免疫一周,采用心脏法采血,离心分离出血清。再与2%～5%SRBC用试管凝集实验和试管溶血实验对其效价进行测定。结果:兔血清中出现了抗体,其中试管凝集实验效价为1∶8,试管溶血实验的效价为1∶32。结论:此免疫血清制备的方法所用的时间相对较短,测定方法简单易行,效果较明显,适用于实验教学和实验研究。     

静脉留置针肝素钠封管液对预防静脉血栓形成的影响

目的 探讨静脉留置针肝素钠封管液与0.9％氯化钠注射液封管液对预防静脉血栓形成的影响.方法 将40例患者按入院的先后顺序分为2组：观察组和对照组,每组20例.观察组采用肝素钠稀释液（含肝素钠25 U·mL-1）进行脉冲式正压封管,对照组采用0.9％氯化钠注射液进行脉冲式正压封管.观察2组静脉血栓形成的情况.结果 观察组有血栓形成发生率显著低于对照组（P＜0.05）.结论 对肾病综合征患者采用肝素钠封管液封管效果较好.     

三型宫内节育器远期效果观察

目的观察不同类型宫内节育器放置5年后的临床效果。方法观察600例门诊放置宫内节育器的育龄妇女,分为三组,其中活性药物缓释含铜及引哚美辛MYCU-IUD 200例,活性带铜T形IUD（TCu-380）200例,母体乐IUD（塑料支架）200例,随访5年。结果 MYCU-IUD、TCu-380A及母体乐IUD妊娠率分别为6%,7.5%和10.5%;因异常出血或腹痛不适取出率分别为3%、4%和4.5%。结论活性药物缓释含铜及引哚美辛MYCU-IUD的临床效果明显优于活性带铜T形IUD和母体乐IUD,为育龄妇女使用宫内节育器的首选。     

VitC、KCl及止血敏对3种交叉配血方法的影响探讨

目的了解药物维生素C（VitC）、氯化钾（KCl）及止血敏对盐水介质、微柱凝胶配血法的影响,并与凝聚胺配血法对比。方法分别在含与不含有特异性抗体两种条件下,加入KCI注射液、酚磺乙胺（止血敏）注射液、VitC注射液、生理盐水,与相应红细胞进行免疫学反应。观察3种药物对不同介质配血方法的影响。结果低浓度的VitC、KCl不影响凝聚胺介质配血,止血敏令试验产生假凝集;高浓度的3种药物可导致试验失败。盐水介质法检测敏感性略低,VitC、KCl对试验的影响不明显,而KCl组随着抗体被稀释,凝集强度减弱甚至于消失。结论3种药物对微柱凝胶配血法均有影响,尤其受KCI影响较大。     

Treatment for the decline of ionized calcium levels during peripheral blood progenitor cell harvesting

BACKGROUND: ACD-A solution containing sodium citrate and citric acid is used as an anticoagulant agent during peripheral blood progenitor cell (PBPC) harvesting, and in rare cases can cause fatal citrate intoxication. The aim of this study was to establish effective methods for stabilizing ionized calcium (ICa) levels during PBPC harvesting. STUDY DESIGN AND METHODS: ICa was measured during 46 apheresis procedures conducted in 26 patients. Four patients in four procedures were infused with calcium gluconate solution before PBPC harvesting; three patients in six procedures were infused with calcium gluconate when symptoms of citrate intoxication appeared; and four patients in five procedures received a continuous infusion. Five patients in five procedures took an isotonic sports drink containing calcium when hypocalcemic symptoms appeared. The ICa level, blood pressure, and pulse rate were measured. RESULTS: ICa declined rapidly from the preapheresis level of 1.081(+/-0.092) mM to 0.937(+/-0.081) mM (13.3%, p < 0.0001) 10 minutes after the start of apheresis and continued to decline until the completion of the procedure. When patients received a continuous infusion of calcium during apheresis, ICa was relatively stabilized. ICa significantly rose (6.1 +/- 3.6%, p < 0.02) within 2 to 5 minutes after oral intake of an isotonic sports drink containing calcium and was maintained within normal range for 31 to 55 minutes. CONCLUSION: An isotonic sports drink containing calcium has a quick stabilizing and a longer maintenance effect on ICa. Thus, we recommend the intake of an isotonic sports drink containing calcium as the easiest and best method for preventing hypocalcemia during apheresis.     

Half-strength citrate CPD and new additive solutions for improved blood preservation. 2. The effect of storage at ambient temperature before component preparation and different means of supplying glucose to the red cells

Summary. In current anticoagulant-additive systems used for the preparation of blood components an early loss of erythrocyte 2,3-bisphosphoglycerate (BPG) during storage is unavoidable. This increases reversibly the oxygen affinity of the haemoglobin of red blood cells (RBC). Using half-strength citrate CPD solution (0·5CPD) as anticoagulant and a red-cell additive solution (RAS 2) containing citrate, adenine, phosphate, mannitol and glucose, it was possible effectively to counteract this undesirable effect without loss of adenine nucleotides. The initial mean BPG level was maintained for 4 weeks and the total adenine nucleotide concentration for 7 weeks. The initial BPG level decreased upon prolonged hold of the whole blood at ambient temperature to 85–90% of normal after 4 h, 70% after 12 h and 50% after 22 h. Both preparation and storage haemolysis were lower in 0·5CPD blood processed into RAS-2-suspended RBC than in CPD blood processed into SAGM-suspended cells.
The possibility of adding glucose to the red cells via the primary anticoagulant instead of the additive solution was tested. The glucose concentration had to be increased to three times that in normal CPD to supply the red cells with sufficient glucose for 6–7 weeks of storage. However, this caused increased haemolysis, particularly after prolonged hold of the whole blood at ambient temperature before component preparation. The results therefore indicate that the glucose supply should be in the additive solution. Less citric acid in the 0·5CPD anticoagulant, an increase in intracellular pH on addition of the new additive solution and supply of phosphate seem to be major factors in explaining the effects. The new combination of anticoagulant and red-cell additive solution is a clear improvement for blood component preparation and storage.     

Citrate anticoagulation and the dynamics of thrombin generation

BACKGROUND: Sodium citrate has been used as an anticoagulant to stabilize blood and blood products for over 100 years, presumably by sequestering Ca(++) ions in vitro. Anticoagulation of blood without chelation can be achieved by inhibition of the contact pathway by corn trypsin inhibitor (CTI). OBJECTIVE: To evaluate the influence of citrate anticoagulation on the performance of blood, platelet-rich and platelet-poor plasma assays. METHODS: Blood was anticoagulated in three ways: by collection into citrate, CTI and citrate with CTI. Plasma was prepared using each anticoagulation regimen. Functional analyses included calibrated automated thrombography, thromboelastography, plasma clotting, the synthetic coagulation proteome and platelet aggregation. Coagulation reactions were initiated with tissue factor-phospholipid and Ca(++) (when indicated). RESULTS: In all cases, citrate anticoagulation resulted in reaction dynamics significantly altered relative to blood or plasma stabilized with CTI alone. Subsequent experiments showed that calcium citrate itself impairs coagulation dynamics. CONCLUSION: Coagulation analyses using blood that has been exposed to citrate and recalcified do not yield reliable depictions of the natural dynamics of blood coagulation processes.     

Comparison of incidence/risk of venous thromboembolism (VTE) among selected clinical and hereditary risk markers: A community-based cohort study

Background

The use of mouse models for the study of thrombotic disorders has gained increasing importance. Methods for measurement of coagulation activation in mice are, however, scarce. The primary aim of this study was to develop a specific mouse thrombin-antithrombin (TAT) ELISA for measurement of coagulation activation and to compare it with two commercially available assays for human TAT complexes. In addition, we aimed to improve methods for mouse plasma anticoagulation and preparation.

Methods and results

First, for the measurement of TAT-complexes in plasma a mouse specific TAT-ELISA was developed using rabbit polyclonal antibodies raised against mouse thrombin and rat antithrombin, respectively. This ELISA detected an increase in TAT levels in a mouse model of endotoxemia. Two commercial human TAT ELISAs appeared to be less specific for mouse thrombin-rat antithrombin complexes. Second, to prevent clotting of mouse blood sodium citrate was either mixed with blood during collection in a syringe or was injected intravenously immediately prior to blood collection. Intravenous sodium citrate completely inhibited blood coagulation resulting in plasma with consistently low TAT levels. Sodium citrate mixed with blood during collection resulted in increased TAT levels in 4 out of 16 plasma samples. Third, heparinase was added to plasma samples after in vivo injection of different heparin doses to test its neutralizing effect. Heparinase neutralized up to a 20 U of heparin/mouse and resulted in accurate APTT and factor VIII determinations.

Conclusion

These procedures and reagents for plasma preparation and coagulation testing will improve studies on thrombotic disorders in mice.     

The effect of sodium citrate in arterial catheters on acid-base and electrolyte measurements

OBJECTIVE: To compare the effects of heparin or sodium citrate used to anticoagulate indwelling arterial catheters on acid-base and electrolyte measurements. DESIGN: Randomized controlled trial. SETTING: Medical-surgical university-affiliated intensive care unit. SUBJECTS: Twenty patients with indwelling arterial catheters. INTERVENTIONS: Patients were randomly allocated to have ten 1-mL aliquots of blood sampled serially from an arterial catheter maintained with either heparin or sodium citrate. A sample then obtained by arterial puncture provided true measurement values. Acid-base and electrolyte measurements of whole blood were obtained from each sample by means of a Coming 860 analyzer. MEASUREMENTS AND MAIN RESULTS: Contamination with sodium citrate lowered ionized calcium and pH but increased glucose and Pco2. Heparin produced negligible effects on those measurements. When sodium citrate was used, reliable measurements were not obtained for ionized calcium, pH, and glucose, even after 9 mL of blood had been discarded. However, reliable P(CO2) measurements were obtained after 2 mL of blood was discarded. CONCLUSIONS: Sodium citrate used to maintain arterial catheters can contaminate blood samples. The result of that contamination can mimic severe hypocalcemia, metabolic acidosis, and mild hyperglycemia. Failure to recognize the effects of sodium citrate on acid-base and electrolyte measurements may lead to changes in treatment that could affect patient outcome adversely.     

高浓度枸橼酸钠与局部肝素体外抗凝在血液净化伴出血倾向患者中运用的效果

目的观察高浓度枸橼酸钠和局部肝素体外抗凝在血液净化伴出血倾向患者中运用的效果。方法 2015年9月,便利抽样法选取在第二军医大学长征医院南京分院肾内科需要行连续性血液净化(continuous blood purification,CBP)治疗的伴出血倾向患者27例为研究对象,按随机数字表法将其分为A、B两组。A组(14例)以高浓度枸橼酸钠抗凝行CBP治疗,B组(13例)以局部肝素体外抗凝行CBP治疗。记录两组患者治疗前后电解质、肌酐、血气及活化部分凝血活酶时间(partial thromboplastin time,APTT),并观察两组滤器及管路凝血情况。结果两组患者均顺利完成CBP治疗,在治疗中均发生滤器及管路Ⅰ级凝血各1例,未出现Ⅰ级以上凝血。A组患者透析后未发生高钠、低钙血症及严重的碱血症,未出现口唇、四肢麻木的低钙表现,患者均存在轻度碱血症,与枸橼酸抗凝呈碱性相关;B组患者在治疗8~12 h时,监测出血时间发现,APTT时间轻度延长。A组和B组患者各实验室指标的变化程度不同。结论对于具有出血倾向患者CBP治疗时,建议首选高浓度枸橼酸钠抗凝,其次是局部肝素体外抗凝。     

两种不同抗凝方法在连续性静脉静脉血液透析滤过中应用的对比研究

目的观察连续性静脉静脉血液透析滤过（CVVHDF）治疗中常规肝素抗凝与体外枸橼酸钠抗凝两种方法的效果与特点。方法多器官功能不全病人70例，按照随机原则分为两组：A组（体外枸橼酸钠抗凝组）（n=37），B组（常规肝素抗凝组）（n=33）行连续性静脉静脉血液透析滤过治疗，记录治疗前、后的血电解质，酸碱度，凝血指标的变化。管路及血滤器的凝血情况和使用时间，并观察治疗中不良反应。结果治疗过程中两组患者生命体征平稳，电解质、血气指标稳定，肌酐、尿素清除效果明显，超滤可达目标值，A组血滤器的凝血情况和使用时间优于B组，A组体内凝血时间不受影响，A组葡萄糖酸钙用量明显大于B组。结论在CVVHDF体外枸橼酸钠抗凝对比常规肝素抗凝更有效和更安全。     

Sodium citrate versus saline catheter locks for non-tunneled hemodialysis central venous catheters in critically ill adults: a randomized controlled trial

Purpose  

Sodium citrate has antibacterial and anticoagulant properties that are confined to the catheter when used as a catheter lock. Studies of its use as a catheter lock in chronic hemodialysis patients suggest it may be efficacious in preventing infection and thrombotic complications. We compared sodium citrate with saline catheter locks for non-tunneled hemodialysis central venous catheters in critically ill adult patients. Primary endpoint was catheter life span without complication.     

高危出血风险患者无肝素透析后肝素盐水封管对凝血指标的影响

目的 通过自身对照比较无肝素透析前、后和封管后30min凝血指标与非透析日基线值的差异,了解高危出血患者无肝素透析后肝素盐水封管对出血风险的影响. 方法 解放军总医院第一附属医院肾内科2011年6～9月间无肝素透析的患者共26例,透析后使用1∶1肝素盐水(3125U/ml)按照导管标记容积封管.事先记录非透析日的血凝指标作为基线值,并于透析开始5min、透析后(封管前),封管后30min分别采血检测血凝指标. 结果 该组患者在上述4个时间点的部分凝血活酶时间(APTT)分别为28.75 (26.60～31.63)、32.65(28.30～54.03)、28.25(26.53～32.03)、47.90 (35.80～199.65),其中透析后(封管前)数据较基线值无差异,但透析开始后5min、封管后30min的数据较基线值明显延长(P=0.0003,0.0001),其中位数比较基线值分别延长达13.6％和66.7％.其他指标如凝血酶原时间(PT)、国际标准化比值(INR),在封管后较封管前也都有明显延长(P≤0.0001). 结论 常规的无肝素透析对凝血各指标影响不明显,但1∶1肝素盐水封管将造成透析开始后短时间内和透析后一定时段内APTT的明显延长,因此,目前采用的肝素盐水封管方式很可能增加出血风险.     

二氧化氯溶液杀灭皮肤真菌效果的检测

以悬液定量杀菌试验,观察二氧化氯溶液加柠檬酸钠与否对须发癣菌与红色毛癣菌的杀灭效果。结果,以含二氧化氯 １２５ ｍｇ／ Ｌ、加与不加 １０％柠檬酸钠的溶液分别对悬液中 ２种真菌作用 ５ ｍｉｎ与１０ ｍｉｎ,杀灭率均达 １００％。