Kidd血型抗原基因定型技术的建立与初步应用

目的 建立简易的Kidd血型抗原基因分型技术.方法 设计2对引物,分别特异性针对Kidd血型基因(SLC14A1)Jka和Jkb抗原的等位基因(JKA和JKB),建立序列特异性引物-聚合酶链反应(PCR-SSP)技术,基因定型Jka和Jkb抗原,初步用于检测192份无偿献血者DNA样品.全部样品均进行血清学红细胞Kidd抗原鉴定.结果 建立的PCR-SSP技术检测192份DNA样品的基因分型结果均与血清学检测结果一致,样品中发现Jk(a+b-)17例(8.854%),Jk(a+b+)125例(65.104%),Jk(a-b+)50例(26.042%),未发现Jk(a-b-)个体.JKA基因频率为0.414 063(41.4%),JKB基因频率为0.585 937(58.6%).结论 成功建立Kidd血型系统抗原基因分型技术,可用于常规检测.     

罕见抗-JK3引起新生儿溶血病及家系Kidd血型基因研究

目的 研究Kidd系统稀有血型Jk(a-b-)产生抗-JK3引起的新生儿溶血病,Jk(a-b-)表型的分子机理及其家系Kidd血型的基因遗传.方法 对患儿血样做新生儿溶血病三项检测:直接抗人球蛋白试验、血清游离抗体和红细胞放散试验;患儿母血样做抗体鉴定及Kidd血型鉴定;患儿父血样做Kidd血型鉴定;采用PCR-SSP方法对患儿及其父母做Kidd血型的基因检测,并对Jk4-11外显子及其侧翼区域扩增后测序,分析其序列信息.结果 患儿为B型RhD阳性、Kidd表型为Jk(a-b+)、基因型为JKB/JKB (IVS5-1);患儿母为O型RhD阳性,Kidd表型为Jk(a-b-)、基因型为Jkb(IVS5-1)/JKb(IVS5-1);患儿父为B型RhD阳性,Kidd表型为Jk(a-b+)、基因型为Jkb3kb;患儿母血浆中存在抗-Jk3,患儿为抗-Jk3引起的新生儿溶血病.结论 患儿母在Jkb等位基因第5内含子3'端保守区剪切结合位点处发生了G到A的突变,导致mRNA转录本中从第6外显子开始缺失,致其Kidd表型为Jk(a-b-),其因妊娠免疫产生了抗-JK3并导致患儿发生新生儿溶血病.     

凝聚胺法在不规则抗体筛检中的应用

目的 探讨凝聚胺法在献血者不规则抗体筛查中的可行性.方法 采用双盲法,分别用抗人球蛋白法和凝聚胺法检测随机抽取的献血者血清标本1 936例.结果 抗人球蛋白法和凝聚胺法分别检出不规则抗体17例和19例,2种方法 检测阳性率差异无统计学意义(P＞0.05).结论 凝聚胺法操作简便、迅速、费用低廉,结果 准确可靠,可以广泛用于献血者不规则抗体筛检工作.     

温州地区献血者Kidd血型基因频率调查与输血风险评估

目的 调查温州地区献血者Kidd血型基因频率的分布,对Kidd血型不配合的输血风险进行评估.方法 采用尿素溶血试验,对36 235例献血者进行Jk（a-b-）稀有血型进行筛选,随机抽取672例献血者的标本采用微量板法进行Kidd血型抗原的鉴定,再用血清学方法对JK（a-b-）表型和可疑的Kidd血型抗原的确认.结果 在36 235例献血者中筛选出5例Jk（a-b-）表型,分布频率为0.014％,基因频率为0.012.672例献血者中,Jka和Jkb的基因频率分别为0.44和0。 56.Jka和Jkb不配合的概率分别为0。 21和0。 16,Jka和Jkb不配合的概率合计为0 .37.结论 Kidd血型系统基因频率符合Hardy-weinberg群体遗传平衡法则;若怀疑为Kidd血型系统抗体时,应采用多种配血方法,以确保输血的安全.     

致敏红细胞Rh血型系统的六种鉴定方法比较

目的:探讨红细胞上的致敏抗体对各种Rh血型系统检测方法的影响.方法:105例新生儿溶血病患儿红细胞被分别用抗球蛋白法、凝聚胺法、盐水法、微柱凝胶法(Rh血型系统测定型)以及抗血清加微柱凝胶法、盐水法加微柱凝胶法(IgG型)进行Rh血型系统检测,同时在45℃抗体释放后再检测.结果:72例直接抗人球蛋白试验阴性患儿的Rh系统血型69例6种方法所获结果一致,3例结果不一致;33例直接抗人球蛋白试验阳性患儿的Rh系统血型21例结果不一致.45℃抗体释放后测定的结果一致,盐水法和微柱凝胶法抗体释放前后的结果一致.结论:红细胞上的致敏抗体达到一定数量可影响抗球蛋白法、凝聚胺法、抗血清加微柱凝胶法和盐水法加微柱凝胶法,而盐水法和微柱凝胶法则影响不大,是一种较好的检测方法.     

微柱凝胶技术筛检输血相关血型不规则抗体的研究

目的:研究微柱凝胶间接抗人球蛋白法在输血前不规则抗体筛查中应用的价值,确保输血安全.方法:用微柱凝胶间接抗人球蛋白试验进行红细胞不规则抗体筛查,阳性样本加用凝聚胺法、传统试管间接抗球蛋白试验做比较.结果:阳性结果35例,阳性率为0.7%,其中温自身抗体8例,同种特异性抗体27例.传统的试管抗人球试验漏检2例II缶床意义抗体,凝聚胺法漏检特异性抗体4例.结论:采用微柱凝胶间接抗球蛋白进行输血前抗体筛选,灵敏度高,结果可靠,操作简便,可取代传统抗人球蛋白法和凝聚胺法,用于临床常规检测.     

微柱凝胶法在输血患者不规则抗体筛检中的应用研究

目的研究微柱凝胶法检测Rh血型抗体的临床应用效果.方法应用凝聚胺法、微柱凝胶抗人球蛋白法进行不规则抗体筛查和鉴定.结果 1 680例患者中经微柱凝胶抗人球蛋白法检测不规则血型抗体11例,阳性率0.65%,而凝聚胺法检测不规则血型抗体5例,阳性率0.30%.结论微柱凝胶抗人球蛋白法在不规则血型抗体检测中灵敏度较高,有利血型抗体的筛查,可以在临床推广.     

凝聚胺法交叉配血2067例检测分析

目的:检查受血者和献血者血清是否存在与对方红细胞不相合的不规则抗体,以保障受血者安全输血。方法:受血者血清与献血者红细胞及献血者血清与受血者红细胞用凝聚胺试剂做交叉配血。结果:2 067例患者中检出Rh不规则抗体5例。其中抗-E 3例、抗-c 1例、抗-cE 1例。结论:Rh血型系统抗-D以外不规则抗体导致配血不合比较多见,这些由输血和(或)妊娠免疫而产生的免疫性抗体能引起迟发性溶血反应,是否也应把E抗原列为常规检测值得探讨。     

宜昌地区献血者Kidd血型分布与输血安全调查

目的 调查宜昌地区献血者Kidd血型人群分布状况,评估Kidd血型不合的输血风险.方法 随机抽取202004/202008月宜昌地区560名献血者血液样本为研究对象,采用2mol/L尿素溶血试验进行Jk(a-b-)表型筛查,微量板法进行Kidd血型抗原的鉴定,再用试管法对可疑的Kidd血型抗原进行确认.结果 560名献...     

新乡地区同种抗体筛选及影响因素分析

目的发现有临床意义的不规则抗体,建立稀有血型库,为患者提供抗原相同的血液,降低不规则抗体的产生,减少或避免同种抗体引起输血反应和新生儿溶血病。方法采用凝集胺、抗球蛋白、菠萝酶等方法,利用抗原抗体特异性反应的原理,对随机无偿献血者6000例和医院随机患者4100例做同种抗体筛选比较。结果检出同种抗体56例(0.55%),女性(38/3690例,1.02%)产生同种抗体比例高于男性(18/6410例,0.28%,P<0.001),产生同种抗体者多有输血史或妊娠史(41/56例,73.2%),Rh抗体(26/56例,46.4%)为多见。结论随机混合O型红细胞做筛选时可能漏检个别抗体,酶法、凝聚胺法和抗球蛋白法体均可能有假阴性或假阳性,对重危患者输血前试验宜结合病史并采用几种方法做抗体筛选。     

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.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

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.