3例Wiskott-Aldrich综合征基因突变的特征分析

目的分析3例Wiskott-Aldrich综合征(WAS)患儿基因型及临床表现,以鉴别诊断要点,拓展诊断方法。方法收集2018年6月-2020年7月在安徽省儿童医院新生儿科接受治疗的3例WAS患儿资料,采用全外显子组测序(WES)方法检测患儿及其父母WAS基因突变情况,Sanger测序验证突变位点。结果 3例患儿主要表现为不同程度的血小板减少,其中病例1、3伴有细菌感染,均存在自身免疫性疾病,符合5分标准,为WAS;病例2符合1分标准,为X连锁的血小板减少症(XLT)。3例患儿WAS基因突变分别位于1、4和10号外显子,均为移码突变。3例患儿父亲均正常,母亲均存在与患儿相应位点的突变,说明母亲为WAS基因突变携带者,突变均遗传自母亲。结论基因检测为诊断WAS的有效方法,通过家系WES(trio-WES)检测WAS基因突变位点可了解突变来源,能够为临床提供优生优育参考,为临床治疗WAS提供新思路。     

15例Wiskott-Aldrich综合征患儿的临床特点及基因突变分析

目的 分析Wiskott-Aldrich综合征(WAS)患儿的临床特征及其与基因突变的相关性.方法 回顾性分析15例WAS患儿的临床资料及WAS基因突变的检测结果.结果 15例患儿均为男性,发病中位年龄40 d(1～365 d),诊断中位年龄12个月(2～132个月),均有误诊病史.WAS评分中位值为3分(2～5分),...     

变性高效液相色谱法在肾上腺脑白质营养不良分子诊断中的应用

目的：目的对4个肾上腺脑白质营养不良（ALD）家系进行基因突变分析。方法：应用变性高效液相色谱（DHPLC）技术检测了4个ALD家系的A／3091基因，并通过核苷酸序列分析确证突变位点。结果：所有患儿的母亲，患儿2、患儿3、患儿4弟弟及其表弟与正常对照的PCR产物混合物。均出现DHPLC洗脱双峰。而正常对照均为单峰，提示相应基因片段存在突变位点。上述出现异源双峰的PCR产物经直接测序．证实了相关突变的存在：患儿1母亲为fs Glu 471突变携带者；患儿2存在S108X突变；患儿3存在R617C突变；患儿4弟弟和表弟均存在A141T突变。结论：DHPLC能有效地检测ABCD1基因突变。并为进—步地基因突变筛查与产前诊断提供依据。     

女性Wiskott- Aldrich综合征一例及其研究进展

Wiskott-Aldrich综合征(WAS)是一种以血小板减少、湿疹和免疫缺陷为特征的X-连锁隐形遗传病,主要由编码WAS蛋白(WASp)的基因突变所致。发病多见于男性,国内外罕有女性患者报道。本文作者报道南京市儿童医院确诊的1例女性WAS。病初患儿因“面色苍白2个月,发现皮肤出血点2 d”入院,病程6年余,伴湿疹、反复感染、自身免疫性贫血、血小板减少。基因分析发现患儿WAS基因1号外显子上携带来源于母亲的杂合无义突变,但因表达正常WAS基因的X染色体发生失活偏移,导致女性患病。本文作者回顾该例患者的临床特征,结合遗传分析并检索国内外相关文献,总结女性WAS的发病模式,旨在呼吁临床医师高度重视女性WAS,及时甄别疑似患者行遗传咨询,提高早期诊断、减少误诊。     

儿童甲状腺激素抵抗综合征临床和遗传特点分析及文献复习

目的 分析儿童甲状腺激素抵抗综合征的临床特点及随访资料,并检测患儿及其父母甲状腺激素受体β(TRβ)基因突变情况,对其遗传特点进行分析.方法 回顾性分析我院2000-2014年确诊为甲状腺激素抵抗综合征患儿的临床资料,并在获得知情同意书的前提下对部分患儿、父母及其部分家系进行外周血DNA的TRβ基因突变位点检测.结果 患儿均于院外诊断为Grave's病或先天性甲状腺功能减退症,存在误诊误治.该病临床症状主要集中在神经系统、心脏症状和听力功能受损.4例患儿学习成绩不良,处于中等以下水平,其中1例患儿智力筛查为中度发育迟滞落后,1例智力发育临界.8例患儿均有心动过速.听力检测提示1例存在听力障碍.6例患儿以发现甲状腺肿大、心率增快就诊.实验室检查以异常的血清游离甲状腺激素升高和不受抑制的促甲状腺激素水平正常或升高为特点.心电图检查3例患儿发现异常,1例表现为加速性房性逸搏心律,2例表现为显著窦性心律不齐,其中1例伴预激综合征.共获得4例患儿及父母TRβ基因检测结果.1例患儿及其母亲TRβ基因第9外显子突变,第1235位碱基由胞嘧啶变为腺嘌呤,导致该位点编码的氨基酸由丙氨酸变为天冬氨酸,但患儿母亲没有相应的临床表现.1例患儿TRβ基因编码区出现第949位点突变,导致第3171位碱基由丙氨酸变为苏氨酸,余2例患儿及父母未发现TRβ基因突变.5例患儿应用溴隐亭治疗,随诊1-5年,5例患儿均心率下降明显,至90-92次/min,甲肿及突眼情况较前好转.结论 甲状腺激素抵抗综合征临床变异较多,临床表现主要为心律失常和智能发育问题,误诊率高.该综合征TRβ基因突变检出率不高,即使基因突变位点相同,表型也不尽相同,是否存在其他基因位点突变尚有待进一步证实.经长期随访观察溴隐亭治疗效果满意,为临床治疗提供可能.     

原发性肉碱缺乏症2例报道及其家系的SLC22A5基因突变检测

目的 探讨原发性肉碱缺乏症的诊断与治疗方案,对2例原发性肉碱缺乏症患儿及其家系行SLC22A5基因检测,确定基因突变位点,为家系提供遗传疾病的咨询.方法 用串联质谱技术对1例疑似患儿进行游离肉碱及多种酰基肉碱检测,对游离肉碱降低的患儿行SLC22A5基因突变检测,确诊PCD,对其姐姐行上述检查.对2例确诊PCD患儿补充左旋肉碱治疗,随访11个月.并对其家系行SLC22A5基因检测.结果 2例确诊PCD患儿,1例为临床患儿,另1例为其姐姐,无明显临床表现.2例患儿均检测到基因突变.2例患儿血游离肉碱水平低于参考值,伴多种酰基肉碱显著降低,均给予补充左旋肉碱治疗,1例治疗2月后症状改善,另1例未曾未发病,血游离肉碱及其他酰基肉碱水平上升至正常.2例患儿SLC22A5 c.760C>T,(p.Arg254X)纯合,致病突变;患儿父母亲SLC22A5基因的c.760C位点检测,发现:均携带c.760C>T,(p.Arg254X)杂合突变.结论 应用串联质谱技术检测血游离肉碱、多种酰基肉碱水平及SLA22A5基因突变检测诊断了2例PCD,均补充左旋肉碱取得较好疗效.SLC22A5基因c.760C>T,(p.Arg254X)突变是本家系中患有PCD的致病突变,用错义突变和剪切改变的分析手段对SLC22A5基因的外显子编码区进行直接测序可为PCD家系提供遗传咨询.     

反复发热伴MEFV基因突变3例临床分析

目的：探讨反复发热伴MEFV基因突变患儿基因型与临床表型关系以及MEFV基因突变在反复发热患儿诊治中的价值。方法：报道3例反复发热患儿经全外基因检测发现伴MEFV基因突变,并对3例患儿临床表现、实验室相关检查及MEFV基因突变位点进行分析。结果：3例患儿均反复发热。病例1,发热间歇期不固定（1～6 d）、发热时伴腹痛、头晕等非特异性症状,多次实验室检查不支持感染性、风湿免疫性及肿瘤相关性疾病,抗感染、非甾体类抗炎药及激素治疗无效。病例2,发热间歇期4周,发热时外周血白细胞总数及中性粒细胞数升高,C反应蛋白(CRP)升高,热退后上述指标恢复正常,生长发育不受影响,多次诊断“化脓性扁桃体炎”。病例3,间歇期为2～8周,实验室检查同病例2,多次诊断“化脓性扁桃体炎”及“扁桃体炎”。3例患儿行全外显子组基因检测,结果均检测出伴MEFV基因突变。病例1患者分别杂合携带c. 1105(exon3)C>T(p. P369S)和c. 1223(exon3)G>A(p. R408Q)突变位点,两个突变均来自母亲,不符合“家族性地中海热（FMF）”遗传学诊断标准,诊断为“发热待查”。病例2患者...     

Wiskott-Aldrich综合征继发系统性红斑狼疮1例报告并文献复习

目的 探讨Wiskott-Aldrich综合征（WAS）继发系统性红斑狼疮（systemic lupus erythematosus, SLE）的临床特征。方法 回顾性分析1例WAS继发SLE患儿的临床资料,并复习相关文献。结果 患儿,男,8月龄即出现血小板减少,用糖皮质激素及静脉丙种球蛋白治疗后,血小板计数未恢复到正常水平。11岁时出现蝶形红斑及双下肢皮疹,抗核抗体、抗ds-DNA阳性,诊断SLE;予免疫抑制治疗后,出现肺部真菌感染、肛周感染等并发症。考虑可能合并免疫缺陷病,行全外显子检测提示WAS基因的半合子突变,EXON 10 c.961C>T,p.Arg321Stop,182,明确诊断为WAS继发SLE。患儿最终因呼吸衰竭死亡。结论 婴儿期起病且免疫治疗效果不佳的男性血小板减少患儿需考虑WAS等遗传性血小板减少症可能,积极进行基因筛查以明确诊断,利于早期干预,改善预后。     

24例Wiskott-Aldrich综合征患儿基因型与临床表现型的关系

目的 分析24例Wiskott-Aldrich综合征(Wiskott-Aldrich syndrome,WAS)患儿基因型与临床表现型关系.方法 收集24例WAS患儿临床资料,进行临床表现型评分.流式细胞术检测患儿外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)、WA...     

先天性肾病综合征1例报告及文献复习

目的 探讨先天性肾病综合征患儿的基因突变情况及其临床特点.方法 分析1例先天性肾病综合征患儿及其父母的临床资料并进行文献复习.取静脉血4 ml,提取基因组DNA,行NPHS1、NPHS2基因检测.结果 患儿血、尿巨细胞病毒(CMV)全项检测均阳性,规律足量抗病毒治疗未见改善,患儿有逐渐加重的重度低蛋白血症及重度水肿,在患儿基因检测中发现了NPHS1的G928A(D310)的杂合突变,发现2种碱基变异,E117K(rs3814995),IVS18,+5G＞A,经比对第1种为单核苷酸多态性,第2种突变为位于内含子的变异(IVS18,+5G＞A)尚未见报道.其父亲、母亲尿检正常,父亲基因检测没有第8外显子G928A (D310)突变,但存在两种碱基变异,E117K (rs3814995),(IVS18,+5G＞A).母亲基因检测结果有第8外显子G928A(D310)的杂合突变,及1种碱基变异,E117K(rs3814995).结论 先天性肾病综合征婴儿存在NPHS1基因突变,当患儿同时存在CMV感染并且抗病毒治疗未见改善时,需进一步行基因检测明确.     

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.