CAT和SPRCA技术在不规则抗体筛查试验中的应用

目的评估柱凝集技术（CAT）、固相红细胞粘附技术（SPRCA）技术与试管PEG-IAT技术在不规则抗体筛查中检测结果的一致性及对不同稀释度阳性样本的检出能力。方法对199例申请输血患者随机样本分别采用CAT、SPRCA与试管PEG-IAT3种技术平行进行不规则抗体筛查试验,分析阳性检出率,对检测结果进行一致性检验（kappa检验）。对4例阳性样本分别采用3种技术进行稀释度研究,比较可检测到红细胞同种抗体的最大稀释度差异。结果 CAT、SPRCA与试管PEG-IAT技术在199例不规则抗体筛查中检测结果完全一致（kappa=1）。SPRCA技术可检测到红细胞同种抗体的最大稀释度高于其他2种技术。结论 3种技术均可用于不规则抗体筛查试验。对于弱反应抗体或低效价抗体的检测,应选择灵敏度较高的检测技术。     

免疫组织化学染色前的准备事项

1 抗体的选择、稀释和保存 1.1 选择抗体应先了解其反应谱和适用条件[包括适用切片(石蜡切片抑或冷冻切片)、稀释度和温育时间等]. 1.2 对于未曾使用过的新抗体,应按照有关说明书的提示,以几个不同的稀释度对适宜检材(已知阳性切片/涂片)进行预试验;根据染色阳性表达的程度和检材背景着色程度,确定用于染色的最适稀释度.     

抗精子抗体的间接免疫荧光测定

本文探索了间接免疫荧光技术检测抗精子抗体的有关问题。输精管结扎术后抗精子抗体的测定,应选择附睾精子作抗原,其最适保存条件是将精子悬液置于-20℃,或用乙醇固定后保存于4℃冰箱中,乙醇固定不仅可增强精子抗原性的显示,而且对其抗原性具有保护作用。本方法所用的附睾精子抗原不须进行其他处理;用精子抗原免疫异种动物,抗精子抗体产生早,效价高,而免疫同种动物则抗原性极弱;抗精子抗体的特异性免疫荧光类型以顶体特异性荧光染色为主,根据不同稀释倍数的待测血清所显示的荧光强度,可判断血清抗精子抗体的效价。     

抗原修复液pH值和抗体浓度对免疫组织化学技术检测PD-L2染色结果的影响

目的:探讨不同浓度抗体和不同pH值的抗原修复液对免疫组织化学反应结果的影响,以提高免疫组织化学染色阳性率.方法:采用3种不同抗体浓度作预处理后,分别在pH值6.0和pH值9.0条件下,进行免疫组织化学SP法检测PD-L2分子在食管磷状细胞癌组织中的免疫组织化学染色效果.结果:PD-L2抗体稀释浓度1∶400的免疫组化染色效果优于抗体浓度1∶800;pH值9.0的抗原修复液处理后的免疫组化染色效果优于pH值6.0的抗原修复.结论:PD-L2抗体稀释浓度1∶400,pH值9.0抗原修复液处理后的免疫组化染色效果最佳,既可提高免疫组织化学反应阳性表达率,又能提高制片质量,是理想的检测PD-L2分子免疫组织化学反应的优化方案.     

不同稀释浓度对狂犬疫苗脂质体免疫原性的影响

目的考察不同稀释浓度狂犬疫苗原液制备狂犬疫苗脂质体冻干粉的体液免疫与细胞免疫效果,筛选出最佳稀释浓度。方法将狂犬疫苗原液分别稀释至40 IU/mL、30 IU/mL、25 IU/mL、20 IU/mL、15 IU/mL、10IU/mL,5 IU/mL后采用冻融-冻干法制备疫苗脂质体冻干粉,复溶后腹腔注射免疫小鼠,通过MTT实验检测淋巴细胞增殖能力以及ELISA实验测定小鼠体液中RV-IgG浓度,筛选出最佳稀释浓度。结果当稀释至15 IU/mL以下时,与原液组比较(P<0.05),有较高的刺激指数SI值(1.3313±0.1082),能够刺激机体产生较高的细胞免疫水平。并且在免疫早期能够刺激机体产生更多的RV-IgG抗体(31.84±2.09) IU/mL。结论将疫苗原液稀释至15 IU/mL时制备的狂犬疫苗脂质体冻干粉诱导的细胞免疫与体液免疫效果最佳。     

SARS病毒抗体效价测定方法的建立及应用

目的 :建立抗SARSIgG和IgM抗体效价定量测定方法。方法 :①在SARS冠状病毒抗体IgG诊断试剂盒和IgM诊断试剂盒的基础上 ,通过选择IgG、IgM标准血清样本 ,作系列稀释 ,建立效价和吸光度标准曲线 ,采用电子表格获得最佳拟合方程式 ,应用于标本血清效价计算。②应用该方法测定 80例康复期SARS患者血清SARSIgG、IgM抗体及其效价。 结果 :①取得了良好的拟合方程式 ,R2 均可大于 0 .99。② 80名康复期SARS住院患者血清中SARS病毒IgG抗体 6 1名阳性 ,效价为 31.9± 4 0 .0 (1.1～ 2 6 4 .0 ) ,偏度系数为 3.6 77;19例阴性。 80名康复期SARS住院患者血清SARS病毒IgM抗体 19名阳性 ,效价为 2 .5 9± 1.35 (1.1～ 5 .5 ) ,偏度系数为 0 .92 7。结论 :①SARS冠状病毒抗体定性诊断试剂盒具有良好的敏感性和特异性 ,在选择IgG、IgM标准血清和采用电子表格得出相应的最佳拟合方程式后 ,可计算出患者血清SARS病毒抗体效价。②SARS病毒抗体阳性的康复期SARS患者血清中的SARSIgG抗体呈偏态分散分布 ,无明显规律可循     

抗角蛋白抗体定量分析对类风湿关节炎诊断及病情评估的探讨

目的:通过观察抗角蛋白抗体(AKA)滴度与临床和实验室指标及手和腕关节X线片分期的关系,研究AKA定量分析在诊断类风湿关节炎(RA)中的临床意义.方法:研究对象分RA组、非RA组,采用间接免疫荧光法检测血清AKA,速率散射比浊法检测类风湿因子(RF);对AKA进行定量分析,观察AKA滴度与临床和实验室指标及手和腕关节X线片分期的关系.结果:RA组AKA阳性者AKA滴度分布以1∶10和1∶20为主;从临床效果进行评价,血清稀释度1∶10为对RA诊断的最佳阳性界值点,且特异度随血清稀释度的增加而增高;AKA阳性RA患者AKA滴度对数值与疾病病程、手和腕关节X线片分期、RF浓度呈正相关.结论:RA患者AKA定量分析结果显示滴度分布以1∶10和1∶20为主;从临床效果进行评价,血清稀释度1∶10为对RA诊断的最佳阳性界值点;血清稀释度越高,特异度越强.AKA滴度与RA患者的病情严重程度和疾病活动度呈正相关.     

不同固定液对细胞骨架荧光染色标记效果的影响

目的 比较不同固定液对细胞骨架荧光染色的差异,选择最佳固定方法.方法 分别采用4%甲醛、2%戊二醛、95%乙醇和4%多聚甲醛/MES 4种固定液,对人肺腺癌细胞H1299进行固定,对其微管和微丝荧光标记后,在共聚焦显微镜下观察其形态结构.结果 4%多聚甲醛/MES对细胞的微管和微丝都有很好的固定作用.4%甲醛、2%戊二醛、95%乙醇对细胞的微丝基本能起到固定作用,而4%甲醛对微管的固定效果略差,2%戊二醛、95%乙醇固定的细胞看不到微管的具体形态结构.结论 细胞骨架的固定中需加入骨架稳定剂,且根据实验目的,选择适当的固定液获得最佳标记效果.     

如何选择合适的病理标本固定液容器

<正>活体组织病理诊断是外科疾病诊断的金标准~([1]),如果组织自溶、结构破坏、细胞变形就可能导致无法诊断~([2])。病理标本固定液一般为甲醛溶液,市售的甲醛溶液为含40%甲醛的溶液,其最佳固定效果为含4%甲醛的溶液,又叫做福尔马林溶液,因此在固定病理标本前应将其稀释10倍,即1份的市售甲醛溶液加9份的水。如果是一些怀疑恶性肿瘤或者恶性肿瘤需做免疫组织化学的,则应选择中性甲醛溶液固定,固定组     

大鼠甲胎蛋白的分离纯化和抗体制备

用聚丙烯酰胺凝胶电泳(PAGE)分离纯化大鼠羊水中的甲胎蛋白(AFP)具有简便、省时的特点。将微量AFP注入淋巴结内,制备高效价的AFP抗体。比较四种固定液对AFP免疫组织化学染色的影响,示丙酮-甲醛固定液能很好保存AFP的抗原性,具有阳性颗粒强、背景染色淡的优点,故是AFP免疫组织化学的最佳固定液。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.