STR基因位点检查在异基因造血干细胞移植中的应用

目的 探讨短串联重复序列(STR)基因位点检查在异基因造血干细胞移植中的应用。方法 采用PCR方法对4例异基因造血干细胞移植、1例非清髓造血干细胞移植的供者和受者移植前、后STR基因进行检测，了解造血干细胞植入情况。结果 4例异基因造血干细胞移植的受者移植后STR基因型与受者移植前STR基因型不同，与供者STR基因型完全相同，提示供者造血干细胞的植入；1例非清髓造血干细胞移植的受者移植后STR基因型表现为患者移植前STR基因型和供者STR基因型的嵌合状态。结论 STR基因位点检查可以用来判断异基因造血干细胞移植的植入情况。     

应用STR多态性对肝豆状核变性家系进行产前基因诊断的研究

目的探讨运用多个STR多态性位点单体型分析对肝豆状核变性携带者进行筛查及产前诊断的方法。方法对6个肝豆状核变性家系进行D13S296、D13S301和D13S316三个位点连锁分析。结果在6个WD家系中共检出患儿2例,携带者3例,正常个体1例。结果均经出生后或引产后脐带血验证,与产前诊断完全相符。结论联合应用多个遗传多态性位点进行连锁分析,方法简便、灵活、可靠.     

联合应用四种分子生物学技术对血友病A家系进行基因诊断

目的最大限度提高血友病A(hemophiliaA,HA)患者及家系成员的基因诊断、携带者检出。方法对22例HA患者和两个家系的成员14人首先采用长距离DNA扩增(LD-PCR)技术,直接检测是否为基因倒位及其携带者;对于非倒位的HA家系依次采用FⅧ基因内的位点BclⅠ(RFLP)、内含子13和22中的STR、FⅧ基因外的DXS52(ST14)位点的多态性进行遗传连锁分析。结果22例HA中12例为重型,8例为中型,2例轻型;12例重型中6例为内含子22倒位,占50%,用该基因信息为一家系2名女性进行了检测均为携带者;联合应用间接遗传连锁分析,对2个家系进行检测,家系C中有1名女性为携带者,家系D中2名女性确定为携带者,1名为正常人。结论联合应用四种分子生物学技术,几乎可以为所有有家族史的HA家系作出基因诊断和携带者检出。     

应用荧光复合扩增技术对河南汉族群体9个STR基因座的多态性研究

目的：调查河南汉族群体的D3S1358、vWA、FGA、D8S1179、D21S11、D18S51、D5S818、D13S317、D7S820等9个STR基因座的基因频率分布,获得群体遗传学数据,研究其法医学应用。方法：用PCR复合扩增和四色荧光自动化检测技术分析9个STR位点的基因型,计算各等位基因的分布频率。结果：获得9个STR位点在河南汉族群体中的等位基因分布频率资料,共检出98个等位基因;家系调查,证明上述位点遗传稳定,符合孟德尔遗传规律。9个STR基因座总鉴别机率（TDP）达99.9999996%。结论：上述9个位点是较理想的遗传标记,可用于法庭科学实践中生物物证的个人识别及亲子鉴定。     

13例Wilson病患者基因8号外显子778密码子碱基突变的SSCP及酶切分析

目的 寻找山东籍Wilson病患者WND基因突变热点,以便易于用SSCP及酶切分析方法检出。使无先证者及独生子女家系易于作出基因诊断,同时可用于杂合子筛查。方法 PCR扩增WND基因8号外显子778密码子区域,SSCP检洲点突变,用MSPI内切酶进一步验证G-T突变的正确性。结果 13例Wilson患者,检出778密码子G-T纯合突变2例,杂舍突变7例,未突变4例,共计26条染色体,突变染色体11条,总染色体突变率为42.3％。结论 WND基因8号外显子,778密码子G-T突变是山东WD患者的一个突变热点。SSCP和酶切分析是诊断此位点突变的便捷方法,可用于WD患者及杂合子检出。     

先天性肾上腺皮质增生症21羟化酶基因突变的研究

目的：检测32例21－OHD患儿CYP 21 3个点突变。方法：采用PCR－ACRS方法检测了32例21－OHD患儿及其家系3个点突变。结果：3例患儿为CYP 21第2内含子656位点突变，2例患儿为CYP 21第4外显子CD 172点突变，3例患儿为CYP 21第8外显子CD 356点突变。患儿为纯合子，家系成员为杂合子。结论：采用PCR－ACRS检测CYP 21点突变方法简便，结果可靠。     

常染色体显性遗传病理性近视家系基因定位分析

目的:通过基因连锁定位分析,探讨原发性病理性近视家系的致病基因与最近报道的病理性近视相关连锁位点15q14、15q25的关系.方法:选择一个连续3代发病的常染色体显性遗传高度近视(ADHM)家系,在15q14、15q25这2个2010年新报道的病理性近视致病基因连锁位点上下游各选取4个多态性微卫星标记物STR进行基因分型,采用两点法进行连锁分析.结果:分析显示在15q14区域内marker D15S165其LOD值为1.42,其附近marker连锁分析结果LOD值均＞0.结论:该家系可能存在与15q14的连锁,需选取密度更高的marker或对此区域内眼部功能相关基因进行测序,以确定该家系致病基因的染色体定位.     

家族性乳腺癌患者亲属BRCA基因突变研究

目的研究河北省家族性乳腺癌患者一级亲属BRCA基因突变情况。方法采用聚合酶链反应-单链构象多态性分析（polymerase chain reaction-single strand conformation polymorphism,PCR-SSCP）和基因测序技术对国内外报告中常见的4个BRCA1/BRCA2突变热点区域（BRCA1：外显子2、11、20;BRCA2：外显子11）进行检测。结果在12个家族性乳腺癌家系46例一级亲属中,发现1个BRCA2基因突变位点（5329insT）和1个核苷酸多态性位点（287G〉C）。结论家族性乳腺癌家系中健康一级亲属突变基因和核苷酸多态性位点携带者存在较大的乳腺癌发病风险。     

病态窦房结综合征家系致病基因分析1例并文献复习

目的：通过全外显子测序（WES）探寻病态窦房结综合征（SSS）家系的致病基因,并探讨其基因突变型与临床表型的相关性。方法：收集在上海市儿童医院就诊的1例SSS病人及其家系成员的临床资料,采集病人及家系成员的外周血,抽提血液DNA,通过WES寻找致病基因,利用Sanger测序在家系中验证可能的致病基因突变,使用致病性预测软件预测基因型与表型关系。结果：将测序结果比对分析,多个生物数据库筛选、过滤,发现SCN5A基因c.999-1G> A位点发生杂合突变,是SSS的可能致病基因,此位点突变在汉族儿童中尚属首次报道。结论：SCN5A基因的c.999-1G> A位点突变是导致散发SSS的致病位点;通过对SSS相关致病基因的研究,对于临床医师精准判断此类病人以及未来尽早进行心脏生物起搏治疗具有重要的临床意义。     

Bcl Ⅰ位点在血友病A携带者检测中的应用

目的探索一种更简便、更特异的方法用于血友病A的基因诊断。方法采用聚合酶链反应-限制性片断多态性方法对8个血友病A家系进行BclI位点检测。结果 BclI位点杂合率为24%(4/17)。可以为其中3个血友病A家系提供诊断信息,信息率为3/8。结论 BclI位点为血友病A基因多态性遗传标志之一,用于血友病A基因诊断有较大价值。     

Prolonged schedule of temozolomide (Temodal) plus liposomal doxorubicin (Caelyx) in advanced solid cancers

Temozolomide (Temodal) is an oral imidazotetrazine. Increased temozolomide exposure and subsequent depletion of O-alkylguanine alkyltransferase may improve the activity of temozolomide. The rationale for investigating temozolomide plus Caelyx is based on their antitumor activity, their formulation and no significant overlapping toxicities. We conducted a study of a prolonged schedule of temozolomide (orally on days 1-7 and 15-21) plus Caelyx (day 1) every 28 days. Twenty-one patients (melanoma n=10, sarcoma n=7 and other n=4) were assigned to four dose levels (DL; temozolomide+Caelyx, mg/m): DL1: 100+30 (n=3 patients), DL2: 100+40 (n=6 patients), DL3: 125+40 (n=6 patients) and DL4: 150+40 (n=6 patients). Dose-limiting toxicities were noted after 2 or more cycles in one patient at DL3 (stomatitis) and one patient at DL4 (grade 4 ANC >/=7 days). Treatment delays and/or dose reductions (due to hematological toxicity) were necessary in five of six patients receiving DL4 compared with one of six patients at DL3, and one patient at DL1 and 2. Thus, the recommended dose was temozolomide 125 mg/m (daily for 7 days every other week) plus Caelyx 40 mg/m (day 1 every 4 weeks). Other toxicities were mild. Antitumor activity was observed in eight patients, including one complete response (melanoma), three partial responses (one melanoma, two sarcomas) and four patients with stable disease (three melanomas, one Ewing), with a duration lasting from 14 to 135+weeks. Two melanoma patients showed tumor stabilization in non-irradiated cerebral lesions. This schedule of temozolomide allowed higher dose intensity (1750 mg/m in 4 weeks) compared to the standard 5-day regimen (1000 mg/m in the same amount of time).     

Correlation of CYP2D6 genotype with perhexiline phenotypic metabolizer status

Perhexiline is metabolized by CYP2D6 and has concentration-related hepatoxicity and peripheral neuropathy. The risk of toxicity is reduced using therapeutic drug monitoring. CYP2D6 genotyping before therapy may allow earlier appropriate dosing. This study aimed to determine whether assessment of CYP2D6 genotype in patients on perhexiline could predict accurately metabolizer status as determined by the perhexiline metabolic ratio (MR). Blood samples from patients stabilized on perhexiline were analysed for CYP2D6 genotype and for concentrations of perhexiline and its hydroxy metabolite. The MR was determined. Of 74 patients, five were poor metabolizers (PM) defined by a MR<0.4, and the remainder were extensive metabolizers (EM). The genotypes were: *1/*1 (n=21), *1/*4 (n=18), *1/*2 (n=12), *1/*3 (n=2), *1/*5 (n=1), *1/*9 (n=2), *1/*10 (n=2), *2/*4 (n=4), *2/*2 (n=3), *4/*41 (n=3), *2/*41 (n=1), *41/*41 (n=1), *4/*9 (n=1), *4/*5 (n=1), *5/*6 (n=1) and *4/*6 (n=1). Allele frequencies were consistent with those reported in population studies. The 3 PMs with the lowest MR were predicted by genotype (*4/*5, *5/*6, *4/*6). The other 2 PMs had intermediate metabolizer genotypes and were on CYP2D6 inhibiting drugs. Amongst the EMs, the highest MR was associated with *1 and *2 allele combinations and the MR was progressively lower with the presence of alleles with intermediate function (*9, *10, *41) followed by alleles with no functional product (*3, *4, *5, *6). Thus, a gene-dose effect was observed. Genotype predicted PM phenotype and also intermediate metabolizers. Determination of CYP2D6 genotype before therapy with perhexiline may help predict perhexiline dose requirements and reduce the risk of perhexiline concentration-related toxicity.     

肝型Wilson病患儿智力结构及心理行为特点分析

目的 研究肝型Wilson病儿童智力结构及心理行为学的特点。方法 选取安徽省中医院脑病中心2013年1月至2016年12月收治的40例肝型Wilson病患儿作为观察组,并选取同期40例健康儿童作为对照组。采用中国修订的韦氏儿童智力测验量表（C-WISC）和Achenbach儿童行为量表分别对两组儿童进行测验,比较两组儿童的智力结构及心理行为的差异。结果 观察组患儿与对照组儿童智力结构比较差异无统计学意义（P>0.05）;观察组患儿在多动、强迫性、交往不良、违纪、攻击性、社交退缩评分均高于对照组,差异有统计学意义（P<0.05）。结论 肝型Wilson病患儿智力总体水平与正常儿童无差异性;而其心理行为存在多方面的异常,主要表现为多动、强迫性、交往不良、违纪、攻击性、社交退缩。     

磁共振胰胆管造影的临床应用

目的：探讨磁共振胰胆管造影（ＭＲＣＰ）的临床应用价值,方法：应用半傅立叶转换自旋回波序列（ＦＡＳＥ）,重Ｔ２加权,对４１例胰胆管疾病患者行ＭＲＣＰ检查。结果：４１例中２４例经手术和病理证实（５８．５％）其中胆管癌９例,壶腹痛６例,胆总管结石４例,胰头癌４例,慢性胰腺炎１例,余１７例未手术,属临床诊断（４１．５％）其中硬化性胆管炎单凭ＭＲＣＰ难以诊断,结论：ＭＲＣＰ可清楚显示胰胆管的形态,对梗阻部位     

Sexually Transmitted Diseases Diagnosed Among Travelers Returning from the Tropics

Objectives.  Data are lacking on the spectrum of sexually transmitted diseases (STDs) diagnosed in returning travelers.
Study Design.  All consecutive travelers consulting our tropical unit between November 1, 2002 and October 31, 2003 were included if they presented within 1 month after their return from the tropics, with mucocutaneous signs suggesting STDs.
Results.  Forty-nine patients (12 women and 37 men; median age 36.4 y, 35 heterosexuals) were included. Four patients had traveled with their usual sexual partner and 45 patients had casual sex while abroad (31 with locals and 14 with other tourists). The main diagnoses were gonococcal urethritis ( n = 18), herpes simplex virus 2 infection ( n = 12), urethritis of undetermined origin ( n = 9), Chlamydia trachomatis infection ( n = 4), primary syphilis ( n = 4), and primary human immunodeficiency virus infection ( n = 2).
Conclusions.  These results illustrate the broad spectrum of STDs contracted by travelers to the tropics. They suggest the need to also inform travelers of the risks of STD and to promote the use of condoms in case of casual sex while abroad.     

Rescue therapy with tacrolimus is effective in patients with severe and refractory inflammatory bowel disease

BACKGROUND: Oral tacrolimus, approved for the prophylaxis of organ rejection in liver or kidney transplants, has been reported to be effective in anecdotal cases of refractory inflammatory bowel disease. AIM: To evaluate the usefulness of low-dose oral tacrolimus in refractory inflammatory bowel disease. METHODS: Thirty-one adult Caucasian patients with steroid-dependent (n = 15) or steroid-refractory (n = 16) inflammatory bowel disease (Crohn's disease, n = 6; ulcerative colitis, n = 23; pouchitis, n = 2) were enrolled. Tacrolimus (0.1 mg/kg body weight per day) was administered orally in 30 patients and initially intravenously in one patient (0.01 mg/kg body weight per day), aiming for serum trough levels of 4-6 ng/mL. The median treatment duration was 12 months (range, 1-137 months). RESULTS: Twenty-eight patients (90.3%) experienced a clinical and laboratory response and 20 (64.5%) went into remission. One ulcerative colitis patient and two Crohn's disease patients did not improve. Three ulcerative colitis patients (9.7%) were colectomized at 1, 12 and 24 months after tacrolimus initiation. In 19 of 23 patients (82.6%) taking steroids, steroids were reduced or discontinued. Side-effects included a temporary rise of creatinine (n = 3, 9.7%), tremor or paraesthesias (n = 3, 9.7%), hyperkalaemia (n = 1, 3.2%), hypertension (n = 1, 3.2%) and an opportunistic infection (n = 1, 3.2%). CONCLUSION: Oral tacrolimus is safe and effective in refractory inflammatory bowel disease.     

Effective Treatment of Neurological Symptoms with Normal Doses of Botulinum Neurotoxin in Wilson’s Disease: Six Cases and Literature Review

Recent cell-based and animal experiments have demonstrated an effective reduction in botulinum neurotoxin A (BoNT/A) by copper. Aim: We aimed to analyze whether the successful symptomatic BoNT/A treatment of patients with Wilson’s disease (WD) corresponds with unusually high doses per session. Among the 156 WD patients regularly seen at the outpatient department of the university hospital in Düsseldorf (Germany), only 6 patients had been treated with BoNT/A during the past 5 years. The laboratory findings, indications for BoNT treatment, preparations, and doses per session were extracted retrospectively from the charts. These parameters were compared with those of 13 other patients described in the literature. BoNT/A injection therapy is a rare (<4%) symptomatic treatment in WD, only necessary in exceptional cases, and is often applied only transiently. In those cases for which dose information was available, the dose per session and indication appear to be within usual limits. Despite the evidence that copper can interfere with the botulinum toxin in preclinical models, patients with WD do not require higher doses of the toxin than other patients with dystonia.     

The E4 allele of apolipoprotein E is associated with increased restenosis after coronary angioplasty

The aim of this study is to investigate the influence of the E4 allele of apolipoprotein E (apo E) on restenosis after percutaneous transluminal coronary angioplasty (PTCA). The subjects were 171 male patients with more than 75% luminal diameter stenotic lesions of the coronary artery who had undergone an elective initial PTCA. The PTCA was successful in 164 patients, 157 of whom completed a prospective 5 month coronary angiography (CAG) follow up to assess the degree of restenosis after their surgery. Patients with previous coronary artery bypass grafting surgery (CABG), 3 vessel disease, complete obstruction or calcified lesions of the coronary artery, cerebro-vascular disease (CVD), arteriosclerosis obliterans (ASO), and renal failure with hemodialysis were excluded, leaving 105 patients in the analysis. Subjects carrying the E4 allele (n = 22, Phenotype E4/2 = 2, E4/3 = 19, E4/4 = 1: E4 group) were well matched with non-carriers (n = 83, Phenotype E2/2 = 0, E3/2 = 4, E3/3 = 79: E3 group) for clinical, and pre-and post-PTCA angiographic features. The restenosis rates were significantly higher in the E4 group than in the E3 group (patient restenosis rate : 59.1 vs 33.7% p < 0.05, lesion restenosis rate: 51.8 vs 30.9% p < 0.05). These results suggest that the E4 allele is associated with a higher restenosis rate after PTCA.     

Comparative clinical study of the efficacy and safety of a S-metoprolol ER tablet versus a racemate metoprolol ER tablet in patients with chronic stable angina

OBJECTIVE: To compare the efficacy and safety of a S-metoprolol extended release (ER) tablet (50 mg) versus a racemate metoprolol ER tablet (100 mg) in the management of angina. METHODS: An open-label, prospective, comparative study in a clinical setting was conducted in Indian patients. Patients (n = 50 in each group) with a history of angina pectoris, with or without hypertension, were administered study medications in a sequential 1:1 manner once daily for 8 weeks. The primary efficacy variable was a mean change from baseline in the number of angina attacks. The secondary efficacy variables were: mean change from baseline in the proportion of patients with no angina attacks, systolic blood pressure, diastolic blood pressure, heart rate, and proportion of blood pressure responders. Number of patients reporting adverse effects (AEs) and severity of AEs in both of the groups were compared. RESULTS: All patients (n = 100) completed the study. In the S-metoprolol group the number of angina attacks (mean +/- SEM) at baseline and after 2, 4 and 8 weeks of therapy were 6.3 +/- 0.8, 3 +/-0.4, 1.8 +/- 0.4 and 0.7 +/- 0.2, respectively. In the metoprolol group these values were 5.8 +/-1, 3 +/- 0.7, 1.4 +/- 0.3 and 0.7 +/- 0.2, respectively. The reduction in the number of angina attacks from baseline was significant (p < 0.0001) in both groups with no between-group difference. The response rate in angina (percentage of patients completely relieved of angina attacks clinically) was greater in the S-metoprolol group (72%) when compared to the metoprolol group (62%) (p > 0.05, NS). Both study groups showed significant (p < 0.0001) reduction in baseline systolic blood pressure (SBP), diastolic blood pressure (SBP) and heart rate (HR) in hypertensive patients and a clinically non-significant (p > 0.05, NS) change in normotensive patients. Among hypertensive patients, the response rate in angina was higher in the S-metoprolol group (74%) when compared to the metoprolol group (61%) (p > 0.05, NS). In the S-metoprolol group four patients reported AEs: fatigue (n = 4), dry mouth (n = 1), dizziness (n = 1), dyspnea (n = 2), and mild rash (n = 1). In the metoprolol group three patients reported AEs: fatigue (n = 2), dyspnea (n = 1) and dizziness (n = 1). No statistically significant difference was detected between the groups in AE frequency/severity. CONCLUSION: In routine clinical practice in the management of angina (with or without coexisting hypertension), S-metoprolol administered at half the dose of the racemate, shows similar efficacy, safety and a trend towards a higher response rate.