Real-time nucleic acid sequence-based amplification using molecular beacons for detection of enterovirus RNA in clinical specimens

Real-time nucleic acid sequence-based amplification (NASBA) using molecular beacon technology (NASBA-beacon) was compared to standard NASBA with postamplification hybridization using electrochemiluminescently labeled probes (NASBA-ECL) for detection of enteroviruses (EV) in 133 cerebrospinal fluid and 27 stool samples. NASBA-ECL and NASBA-beacon were similar in sensitivity, detecting 55 (100%) and 52 (94.5%) EV-positive samples, respectively. There were no false positives. Both NASBA assays were significantly more sensitive than culture. Real-time NASBA-beacon reagents and equipment rental were more expensive than those for NASBA-ECL; however, time to result was shortened by 1.5 h, hands-on time was reduced by 25 min, and the assay was much simpler for technologists to learn and perform.     

Growth hormone benefits children with 18q deletions

Most individuals with constitutional deletions of chromosome 18q have developmental delays, dysmyelination of the brain, and growth failure due to growth hormone deficiency. We monitored the effects of growth hormone treatment by evaluating 23 individuals for changes in growth, nonverbal intelligence quotient (nIQ), and quantitative brain MRI changes. Over an average of 37 months, the treated group of 13 children had an average nIQ increase of 17 points, an increase in height standard deviation score of 1.7, and significant change in T1 relaxation times in the caudate and frontal white matter. Cognitive changes of this magnitude are clinically significant and are anticipated to have an effect on the long-term outcomes for the treated individuals.     

Skeletal changes in epidermal nevus syndrome: does focal bone disease harbor clues concerning pathogenesis?

Epidermal nevus syndrome (ENS) is a rare, sporadic, congenital disorder of unknown etiology featuring a complex and highly variable phenotype that can include focal or generalized skeletal disease. We describe a young man with ENS manifesting right-sided linear skin lesions, generalized weakness, diffuse osteopenia associated with hypophosphatemic rickets, and distinctive focal bone lesions ipsilateral to the skin findings. Review of the literature concerning ENS-associated skeletal disease suggested such focal bone defects are fibrous dysplasia, but our patient did not have the typical radiographic or histopathologic findings of fibrous dysplasia. Nevertheless, his circulating fibroblast growth factor 23 (FGF-23) level was elevated, likely functioning as a "phosphatonin," yet no activating mutations in GNAS previously reported in fibrous dysplasia or McCune-Albright syndrome were detected in his leukocytes or affected skin. We postulate that the focal skeletal disease, although different than fibrous dysplasia, may be a source of FGF-23 in ENS.     

Discriminatory power and reproducibility of novel DNA typing methods for Mycobacterium tuberculosis complex strains

In recent years various novel DNA typing methods have been developed which are faster and easier to perform than the current internationally standardized IS6110 restriction fragment length polymorphism typing method. However, there has been no overview of the utility of these novel typing methods, and it is largely unknown how they compare to previously published methods. In this study, the discriminative power and reproducibility of nine recently described PCR-based typing methods for Mycobacterium tuberculosis were investigated using the strain collection of the interlaboratory study of Kremer et al. This strain collection contains 90 M. tuberculosis complex and 10 non-M. tuberculosis complex mycobacterial strains, as well as 31 duplicated DNA samples to assess reproducibility. The highest reproducibility was found with variable numbers of tandem repeat typing using mycobacterial interspersed repetitive units (MIRU VNTR) and fast ligation-mediated PCR (FLiP), followed by second-generation spoligotyping, ligation-mediated PCR (LM-PCR), VNTR typing using five repeat loci identified at the Queens University of Belfast (QUB VNTR), and the Amadio speciation PCR. Poor reproducibility was associated with fluorescent amplified fragment length polymorphism typing, which was performed in three different laboratories. The methods were ordered from highest discrimination to lowest by the Hunter-Gaston discriminative index as follows: QUB VNTR typing, MIRU VNTR typing, FLiP, LM-PCR, and spoligotyping. We conclude that both VNTR typing methods and FLiP typing are rapid, highly reliable, and discriminative epidemiological typing methods for M. tuberculosis and that VNTR typing is the epidemiological typing method of choice for the near future.     

Diminished neo-antigen response to keyhole limpet hemocyanin (KLH) vaccines in patients after treatment with chemotherapy or hematopoietic cell transplantation

Relapse is the most common cause of treatment failure for advanced cancer, even those treated with autologous hematopoietic cell transplantation (HCT). Effective tumor-specific immunotherapy may decrease relapse, however, this will fail if the immune system is unable to respond. We developed a strategy to test immune responses with a single injection of the bona fide neo-antigen KLH. The model was first tested in 37 normal volunteers using three KLH vaccines: Intracel KLH, Biosyn KLH, and Biosyn KLH + adjuvant. Despite finding the immunogenic epitope conserved in both products, intact Intracel KLH induced a better response compared to a purified 350/390 kDA subunit of KLH contained in the Biosyn KLH product. Addition of a synthetic oil adjuvant (Montanide ISA51) restored the response to a single injection of Biosyn KLH. A quantitative readout measured by a KLH-specific cellular and humoral response with isotype switching 1 month after KLH vaccination was established. To test the integrity of the adaptive immune response in cancer patients, we vaccinated 14 patients post-HCT and 19 patients with advanced cancer with KLH vaccines that elicited a 100% response rate in normal volunteers. In marked contrast to normal subjects, both responses were significantly impaired up to 16 months after autologous HCT with an intermediate response in advanced cancer patients. KLH vaccines are safe and require only a single injection to test neo-antigen responses providing an optimal platform for definitive testing of strategies to improve diminished immune recovery after chemotherapy or post-HCT.     

转染survivin反义mRNA对Jurkat淋巴瘤细胞生长和化疗敏感性的影响

目的 研究转染survivin反义mRNA对Jurkat淋巴瘤细胞生长的影响以及转染后淋巴瘤细胞对化疗药物的敏感性。方法 构建survivin反义mRNA真核表达质粒pcDNA3．1-反义（As）survivin;利用脂质体转染法将其转入高表达survivin mRNA T淋巴母细胞淋巴瘤Jurkat细胞系,用逆转录聚合酶链反应（RT—PCR）、免疫组织化学SP法、Western印迹法检测细胞中survivin表达;用细胞计数、流式细胞术（FCM）检测其细胞生长曲线、细胞凋亡指数,并进行光镜、电镜形态学观察;并对转染pcDNA3．1-Assurvivin前后Jurkat细胞分别加入4-羟基-环磷酰胺（CTX）、甲氨蝶呤（MTX）72h后,常规MTT检测细胞存活率。结果 RT—PCR检测转染pcDNA3．1-Assurvivin后48h、5和6周Jurkat细胞survivin mRNA表达,发现survivin mRNA表达皆低于对照组;转染后survivin蛋白表达也明显降低。转染pcDNA3．1-Assurvivin后Jurkat细胞生长倍增时间（52h）明显延长;用FCM检测细胞凋亡发现,转染pcDNA3．1-Assurvivin后Jurkat细胞凋亡指数[20．2％（48h）]明显高于对照组（转染空质粒和未转染组,2．1％和1．3％）;5和6周为6．2％和6．8％,明显高于未转染细胞（1．3％和1．0％）。光镜、电镜观察见转染细胞出现较多凋亡细胞及一些变性肿胀细胞;MTT检测结果显示Jurkat细胞转染前后,经化疗药物4-羟基-环磷酰胺和甲氨蝶呤作用,转染细胞的抑制率明显大于未转染组,差异有统计学意义（P〈0．05）。结论 survivin基因对Jurkat细胞系的生长起着重要的作用,抑制survivin基因表达在T淋巴母细胞淋巴瘤治疗中可能有重要的意义,该基因似可能作为治疗的靶点。     

儿童原发性侵袭性真菌病5例分析

目的总结儿童原发性侵袭性真菌病的临床特点,帮助早期诊断及改善预后。方法回顾分析2009年12月至2010年10月住院诊断原发性侵袭性真菌病的5例儿童的临床资料。结果全部患儿均有高热、肺部影像学改变及肝功能异常。痰培养3例阳性,1,3-β-D-葡聚糖(BG)检测2例阳性,血培养2例阳性,骨髓培养2例阳性,组织病理学检查1例阳性。均确认为念珠菌感染,给予氟康唑治疗后痊愈。结论儿童原发性侵袭性真菌病临床表现缺乏特异性,念珠菌感染常见,需结合影像学及实验室检查进行诊断,氟康唑治疗有效。     

Androgenkonzentrationen und kardiovaskuläre Risiken bei der Frau

Low testosterone concentrations in men are associated with a higher cardiovascular risk factor burden. Also in women, cardiovascular risk increases after menopause. The aim of this work was to analyze potential associations between androgen concentrations in women and cardiovascular risk factors, including clinical correlates and sleep, metabolic syndrome (MetS), type 2 diabetes mellitus (T2DM), as well as subclinical and clinical cardiovascular disease (CVD) and mortality. Data from 2000 women of the population-based, longitudinal Study of Health in Pomerania (SHIP) and multivariable regression models were analyzed. Cardiovascular risk factors including obesity, hypertension, dyslipidemia, and poor sleep quality were associated with androgen concentration. Furthermore, cross-sectional and long-term analyses yielded inverse associations between sex hormone-binding globulin (SHBG) and MetS as well as T2DM. No independent, significant associations between androgen concentrations and incidence of CVD, CVD mortality, or all-cause mortality were detected. In this epidemiological study, clinical correlates of androgens in women and associations between androgens and sleep were identified. Associations between androgens, subclinical CVD, and mortality confirmed the relative impact of cofactors. SHBG might represent a potential biomarker for cardiovascular risk factors, especially among postmenopausal women.     

血尿酸水平和儿童阻塞性睡眠呼吸暂停/低通气综合征相关性的临床研究

目的 探讨阻塞性睡眠呼吸暂停/低通气综合征(OSAHS)儿童血尿酸(UA)水平变化.方法 2008年9月-2010年3月,经多道睡眠监测确诊的138例OSAHS儿童纳入研究组,另有65名副耳及上睑下垂的儿童为正常对照组,另外,按照呼吸暂停低通气指数(AHI)或阻塞性呼吸暂停指数将研究组儿童分为轻度组、中度组、重度组3组,再将研究组和对照组按照体质量指数(BMI)各自再分成超重和肥胖组、非超重和肥胖组,比较各组间晨起空腹血UA水平.再分别进行研究组伴或不伴超重和肥胖儿童血UA水平与AHI、BMI、氧减饱和指数、最低血氧饱和度(LSaO2)及血氧饱和度低于0.92时间所占睡眠总时间百分比之间的相关性分析.结果 (1)研究组与对照组之间(z=-0.443)、研究组OSAHS不同病情(x2=1.241)之间的血UA水平差异无统计学意义(P＞0.05).(2)超重和肥胖儿童血UA水平:无论研究组[273.0(238.3～357.3)]还是对照组[298.0(253.0～336.0)],均高于非超重和肥胖儿童[研究组246.5(215.8～300.0),对照组266.0(224.0～303.3)],差异均有统计学意义(z=-2.084,-2.214,P＜0.05).(3)伴或不伴超重和肥胖的研究组血UA水平与AHI、BMI、氧减饱和指数、LSaO2及血氧饱和度低于0.92时间所占睡眠总时间百分比等指标之间无相关性.结论 与对照组相比,研究组患儿血UA水平无明显升高,两组中超重和肥胖者血UA水平高于非超重和肥胖者,儿童血UA水平与AHI、BMI、氧减饱和指数、LSaO2及血氧饱和度低于0.92时间所占睡眠总时间百分比等指标之间无相关性.伴超重和肥胖的OSAHS儿童及对照组儿童的血UA水平显著高于非超重和肥胖的OSAHS儿童及对照组儿童.

Abstract：

Objective To explore the level of serum uric acid (UA) in children with obstructive sleep apnea/hypopnea syndrome (OSAHS). Method Between Sep. 2008 and Mar. 2010, 138 children with OSAHS were enrolled in study group. Sixty-five children with accessory auricle or ptosis of upper lid were enrolled into the control group. Furthermore, according to apnea/hypopnea index (AHI) or obstructive apnea index ( OAI ) the study group was further divided into three subgroups ( mild, moderate and severe group). At last, the study group and control group were divided into two groups according to the body mass index (BMI), separately. The fasting serum UA level was compared among the different groups. Then the correlation between the serum UA level and AHI, BMI, oxygen desaturation index, least arterial oxygen saturation ( LSaO2 ) and the percentage of total sleep time with arterial oxygen saturation ＜ 0. 92 was also analyzed in OSAHS children with or without overweight and obesity respectively. Result The difference of serum UA level between the study group and control group ( z = - 0. 443 ), and the difference among the three groups (x2 = 1. 241 ) was not significant(P ＞0. 05). The serum UA level in overweight and obese children[study group, 273.0 (238.3 - 357. 3 ); control group, 298.0 ( 253.0 - 336. 0)]was significantly higher than that in children with normal BMI[study group, 246. 5(215.8 -300. 0); control group, 266. 0 (224. 0 - 303.3 )](z = - 2. 084, - 2. 214, P ＜ 0. 05 ). That serum UA level did not correlate with the above index of OSAHS was observed in children with or without overweight and obesity in study group ( P ＞0. 05 ).Conclusion Findings of higher serum UA level were not observed in children with OSAHS. There was no correlation between serum UA level and the above indices of OSAHS. The serum UA level in overweight and obese children was significantly higher than that in children with normal BML.     

Acute systemic immune activation following vaginal exposure to staphylococcal enterotoxin B--implications for menstrual shock

Menstrual toxic shock syndrome (mTSS) is an acute systemic inflammatory disease associated with the superantigenic exotoxin, toxic shock syndrome toxin (TSST)-1, produced by Staphylococcus aureus and the use of high absorbency tampons. Even though S. aureus is capable of elaborating several other superantigenic exotoxins, only TSST-1 has been implicated in the pathogenesis of mTSS possibly because most other superantigenic exotoxins are known enterotoxins. Nonetheless, we have shown recently that one of the enterotoxigenic staphylococcal superantigens, staphylococcal enterotoxin B (SEB), can cause robust systemic immune activation following exposure through non-enteric mucosa, including nasal or conjunctival routes. In a similar manner, we show here that vaginal administration of SEB in HLA class II transgenic mice can cause robust systemic immune activation characterized by profound elevation of proinflammatory cytokines in the serum, activation and expansion of SEB-reactive CD4(+) and CD8(+) T cells in peripheral lymphoid organs and SEB-induced deletion of immature thymocytes. Vaginal administration of SEB also caused leukocytic infiltration in major organs, such as liver and lung, reminiscent of human toxic shock syndrome. Systemic immune activation following vaginal superantigen delivery was independent of the stage of the estrus cycle in the mouse. Using HLA class II transgenic mice, we have shown that exposure to SEB through the vaginal canal can cause robust systemic immune activation. SEB could thus play a role in the pathogenesis of mTSS.