Use of multiple displacement amplification to amplify genomic DNA before sequencing of the alpha and beta haemoglobin genes

AIMS: To evaluate the technique of multiple displacement amplification (MDA) for whole genome amplification from small volume blood samples before sequencing in a clinical test to identify haemoglobin gene mutations. METHODS: Phage phi29 DNA polymerase was used to perform MDA, starting with either 1 micro l of blood or 1 ng of previously isolated blood DNA from 23 patients. The amplified products were then evaluated using a clinical test that involves sequencing the haemoglobin genes to detect mutations. The results were compared with the current clinical test method that uses genomic DNA isolated using column based technology. RESULTS: The MDA technique produced large quantities (theoretically approximately 2 mg) of DNA. The amplification procedure was extremely easy and took about four hours (less than one hour of hands on technician time and three hours for amplification). When MDA products were used in the same clinical test protocol as genomic DNA isolated using column technology, there was 100% concordance for detection of a variety of point mutations in the alpha1, alpha2, and beta globin genes. CONCLUSIONS: The MDA technique is useful for overcoming the problem of insufficient genomic DNA in clinical specimens requiring haemoglobin gene sequencing and could be useful for other clinical applications.     

3-(1H-pyrrol-2-yl)-2-oxazolidinones as novel monoamine oxidase type A inhibitors

A novel series of 5-substituted-3-(1H-pyrrol-2-yl)-2-oxazolidinones 2a-s has been described as pyrrole analogues of toloxatone and befloxatone, two phenyl-oxazolidinones active as anti-MAO agents and used in antidepressant therapy. Tested against MAO-A and MAO-B enzymes, the majority of 2a-s show highly potent inhibitory effect against the A isoform of the enzyme, with Ki values in the range 0.52-0.004 microM, whilst their anti-MAO-B activity is considerably lower (Ki = >100-0.5 microM). Structurally, 2a-s differs for the substituent inserted at the C5 position of the 2-oxazolidinone ring (hydroxymethyl (2a-d), methoxymethyl (2e-h), azidomethyl (2i-l), methylaminomethyl (2m-p), and aminomethyl (2q-s)), and the size of the alkyl chain at the pyrrole N1 position (methyl, ethyl, allyl, or benzyl). As a rule, apart from the C5 substitution, the bulkier is the alkyl group at the pyrrole-N1, the lower is the anti-MAO-A activity of the compounds, being the N1-methyl derivatives 2a, 2e, 2i, and 2q among the most potent (K(iMAO-A) = 0.087-0.004 microM) and A-selective (A-selectivity ratio: >11,111-41) compounds in this series. Exceptions are represented by the N1-benzyl derivative 2d (K(iMAO-A) = 0.009 microM) and the N1-allylpyrrole 2o (K(iMAO-A) = 0.04 microM). In comparison with the reference drugs, these highly active derivatives are more potent than toloxatone, slightly less potent than befloxatone, and several times more A-selective than both the references. Such results indicate that 2a-s may represent a new promising series of antidepressant agents.     

血清、尿液β2-m对Graves'病患者检测的临床意义

目的：探讨血清、尿液β2-微球蛋白（β2-mieroglobulin，β2-m）对毒性弥漫性甲状腺肿（graves diseases，Graves）患者β2-m含量与甲状腺激素的关系，了解其变化情况及临床意义。方法：采用放射免疫分析和化学发光免疫分析测定了82例Graves病初发未治疗组与同一组经治疗的69例缓解组及13例未缓解组。并和40例健康对照组进行血清、尿液β2-m含量与FT3、FL4、TGA、TMA血清检测。结果：初发组与未缓解组β2-m含量与FT3、FT4含量显著高于健康对照组（P〈0．01）。经治疗组β2-m含量与FT3、FT4含量同步缓解而下降至正常，与未治疗组对比（P〈0．01）。对比健康正常组差异无显著性（P〉0．05）。82例Graves病初发组TGA阳性率为48．7％（40／82），TMA阳性率为53．66％（44／82）。未缓解组TGA阳性率为15．38％（2／13），TMA阳性率为23．07％（3／13）。缓解组TGA、TMA仍有13．04％、15．94％阳性。结论：甲状腺疾病患者β2-m含量与甲状腺激素有密切关系，提示β2-m可作为Graves病疗效观察、病情变化及预后判断的一项辅助诊断指标。     

Pathologic study and clinical significance of Hürthle cell papillary thyroid carcinoma.

Hürthle cell papillary thyroid carcinoma is a variant of papillary thyroid carcinoma (PTC). Its pathologic and clinical significance has not been well documented. The authors studied the relative incidence of Hürthle cell PTC and the relationship of Hürthle cell PTC to other variants of thyroid carcinoma. Three hundred eighty consecutive cases of thyroid carcinoma were reviewed to identify cases with focal or extensive areas of Hürthle cell PTC, classic PTC, Hürthle cell carcinoma (ie, non-Hürthle cell PTC), and follicular carcinoma. In addition, the status of lymphoid infiltrate in the tumor, stromal invasion with desmoplastic reaction, vascular invasion, and distant and lymph node metastasis were noted by microscopic examination, review of clinical charts, or both. A total of 24 (HCs) and 42 PTCs with Hürthle cells were identified. The latter category was divided into pure Hürthle cell PTC or extensive Hürthle cell (HPTC) (28 cases) and PTC or Hürthle cell carcinoma with focal areas of Hürthle cell PTC (14 cases). The Hürthle cell PTC/Hürthle cell carcinoma ratio was lower than that of PTC/follicular carcinoma (39:289) (P = 0.001). Follicular or solid structures were present in all HPTCs. HPTCs were associated with frequent stromal intrathyroid and extrathyroid invasion, but they tended to have a lower rate of lymph node metastasis (8/28) compared with classic PTC with stromal invasion (108:200) (P = 0.12) and a lower rate of distant metastasis (2:28) compared with Hürthle cell carcinoma (15:24) (P = 0.02) or follicular carcinoma (13:39) (P = 0.04). Warthin-like Hürthle cell PTC (10 cases) was associated with extrathyroid invasion in five cases. In Hürthle cell PTC associated with tall cell variant (10 cases), areas of gradual transition between Hürthle cell PTC and tall cell variant were identified. The latter variant showed the highest rate of extrathyroid stromal and vascular invasion with distant metastasis and patient death compared with all Hürthle cell PTCs and classic PTCs. In conclusion, Hürthle cell PTC is frequently associated with tall cell variant. It has a higher potential for extrathyroid invasion than classic PTC and has vascular invasion and distant metastasis characteristics intermediate between those of classic PTC and Hürthle cell carcinoma with or follicular carcinoma. Hürthle cell PTC tends to show a greater likelihood of extrathyroid invasion when associated with Warthin-like features and tall cell variant PTC, and higher vascular invasion and distant metastasis when associated with tall cell variant.     

Systematic review of accelerometer-based methods for 24-h physical behavior assessment in young children (0–5 years old)

Background

Accurate accelerometer-based methods are required for assessment of 24-h physical behavior in young children. We aimed to summarize evidence on measurement properties of accelerometer-based methods for assessing 24-h physical behavior in young children.

Methods

We searched PubMed (MEDLINE) up to June 2021 for studies evaluating reliability or validity of accelerometer-based methods for assessing physical activity (PA), sedentary behavior (SB), or sleep in 0–5-year-olds. Studies using a subjective comparison measure or an accelerometer-based device that did not directly output time series data were excluded. We developed a Checklist for Assessing the Methodological Quality of studies using Accelerometer-based Methods (CAMQAM) inspired by COnsensus-based Standards for the selection of health Measurement INstruments (COSMIN).

Results

Sixty-two studies were included, examining conventional cut-point-based methods or multi-parameter methods. For infants (0—12 months), several multi-parameter methods proved valid for classifying SB and PA. From three months of age, methods were valid for identifying sleep. In toddlers (1—3 years), cut-points appeared valid for distinguishing SB and light PA (LPA) from moderate-to-vigorous PA (MVPA). One multi-parameter method distinguished toddler specific SB. For sleep, no studies were found in toddlers. In preschoolers (3—5 years), valid hip and wrist cut-points for assessing SB, LPA, MVPA, and wrist cut-points for sleep were identified. Several multi-parameter methods proved valid for identifying SB, LPA, and MVPA, and sleep.

Despite promising results of multi-parameter methods, few models were open-source. While most studies used a single device or axis to measure physical behavior, more promising results were found when combining data derived from different sensor placements or multiple axes.

Conclusions

Up to age three, valid cut-points to assess 24-h physical behavior were lacking, while multi-parameter methods proved valid for distinguishing some waking behaviors. For preschoolers, valid cut-points and algorithms were identified for all physical behaviors. Overall, we recommend more high-quality studies evaluating 24-h accelerometer data from multiple sensor placements and axes for physical behavior assessment. Standardized protocols focusing on including well-defined physical behaviors in different settings representative for children’s developmental stage are required. Using our CAMQAM checklist may further improve methodological study quality.

PROSPERO Registration number

CRD42020184751.

     

用层析法研究X—CT的论证问题

为改善技术论证的定量程度，作者用层析法对Ｘ－ＣＴ的选型操作进行了研究，并提出此方法可推广应用于一般医院的大型装备。     

偏头痛患儿的大脑血流动力学变化

目的　研究偏头痛患儿的脑动脉血流动力学改变 ,探讨其在小儿偏头痛发生中的作用。方法　应用经颅多普勒 (TCD)技术 ,检测 90例 7～ 14岁头痛间歇期的偏头痛患儿和 80例健康同龄儿童脑动脉血管血流动力学指标。结果　 90例偏头痛患儿中脑动脉血流动力学异常者 77例 (86 % ) ,其中 6 0例 (6 7% )流速增快 ,9例 (10 % )减慢 ,与对照组相比 ,差异具有显著性 (P <0 0 1) ;90例中 2 3例 (2 6 % )存在双侧同名动脉血流速度不对称 ,其中伴有血流速度异常者15例 ,血流速度正常者 8例 ,差异具有显著性 (P <0 0 1)。结论　脑动脉血流动力学因素异常改变 ,在小儿偏头痛的发生中具有重要作用。尤其值得注意的是 ,双侧同名动脉流速不对称的改变 ,在小儿偏头痛的发生中同样可能起着重要的作用。     

两种不同血管紧张素ⅡⅠ型受体拮抗剂治疗轻中度原发性高血压的临床研究

目的:探讨缬沙坦和氯沙坦治疗轻中度原发性高血压的临床疗效及安全性。方法:117例轻中度原发性高血压患者随机分为两组,缬沙坦(治疗)组59例,80～160mg/d,口服,疗程半年;氯沙坦(对照)组58例,50～100mg/d,口服,疗程半年。治疗前后测偶测血压、动态血压、肝肾功能、血脂、血糖、血尿酸,血浆肾素活性(PRA)、血管紧张素Ⅱ(AngⅡ)醛固酮(ALd)、内皮素(ET)等,并观察心脑事件发生情况。结果:治疗组总有效率74.6％(44/59例),降压幅度26.2/15.1mmHg,谷/峰(T/P)比值:收缩压(SBP)0.28、舒张压(DBP)0.78,与对照组总有效率74.1％(43/58例),降压幅度25.9/15.8mmHg,T/P比值:SBP 0.81、DBP 0.79均相似(P均>0.05)。两组治疗后PRA、AngⅡ均升高(P均<0.05);Ald、ET均降低(P均<0.05),但组间对照无显著性差异(P>0.05)。血尿酸也明显下降(P均<0.05),且基础值越高,降幅越大,对于治疗前高尿酸血症患者,对照组降压大于治疗组(P<0.05)。半年随访,两组心脑血管事件发生率相似(P>0.05),治疗组不良么应轻微。结论:缬沙坦是一种安全、作用持久、稳定、耐受性好、用药方便的治疗轻中度原发性高血压的新型AngⅡⅠ型受(AT1)拮抗药物之一,值得临床推广应用。