全血细胞自动分析新技术进展

既往血常规检验靠手工操作进行，费工费时，近十年来，各种自动血液分析仪（ＡＨＡ）的迅速发展和应用极大地改变了这种状况，根据电阻变化，激光散射，射频，组织化学和流式细胞仪诸原理设计制造的新式ＡＨＡ可提供包括白细胞分类（ＷＢＣＤＣ）红细胞分化宽度（ＲＤＷ），血红蛋白分布宽度（ＨＤＷ）和血小板分布宽度（ＰＤＷ）在内的全血细胞（ＣＢＣ）多参数分析，应用流式细胞仪技术的ＡＨＡ或专用网织红细胞Ｒｅｔ计数仪还可作     

Validation of mitochondrial DNA sequencing for forensic casework analysis

Two sets of studies were performed to evaluate the forensic utility of sequencing human mitochondrial DNA (mtDNA) derived from various tissues and amplified by the polymerase chain reaction (PCR). Sequencing was performed on a Perkin-Elmer/Applied Biosystems Division (PE/ABD) automated DNA sequencer (model 373A). The first set of experiments included typical validation studies that had previously been conducted on forensic DNA markers, such as: chemical contaminant effects on DNA from blood and semen and the effect of typing DNA extracted from body fluid samples deposited on various substrates. A second set of experiments was performed strictly on human hair shafts. These studies included typing mtDNA from hairs that were: (1) from different body areas, (2) chemically treated, (3) from deceased individuals, and (4) deliberately contaminated with various body fluids. The data confirm that PCR-based mtDNA typing by direct automated sequencing is a valid and reliable means of forensic identification.     

Recent progress in radionuclide therapy

Therapeutic use of radionuclides includes 131I for thyroid cancer and hyperthyroid Graves' disease, 89SrCl3 for metastatic bone tumors, 131I-MIBG for malignant pheochromocytoma and neuroblastoma, and radioimmunotherapies. 131I is concentrated in 60-70% of metastases from differentiated thyroid cancer following total thyroidectomy. Radioiodine uptake in metastatic lesions is greater in younger patients than in older ones. Hypothyroidism is often mild or even absent in patients with a large amount of tumor tissue, indicating that thyroid hormones produced by highly differentiated tumors compensate partially or even completely for hypothyroidism following total thyroidectomy. Adequate uptake of 131I has been reported to be associated with significant reduction in the size and number of metastases, and with lower recurrence and higher survival rates. Other favorable factors for longer survival are younger age, well-differentiated histological type, small disease extent, and early discovery of metastases. Older patients with extensive metastases and/or bulky tumor masses in the bone have a poor prognosis. Therefore, it is important to discover metastases as early as possible, when patients are still young. Long-term follow-up with periodic thyroglobulin measurements and imaging studies is strongly recommended. In Japan, 131I treatment for Graves' disease is performed only in selected patients in whom antithyroid drugs cannot be used because of side effects or not effective, considering the high prevalence of permanent hypothyroidism. 89SrCl3 is useful for reducing pain due to bone metastases of malignant tumors. 131I-MIBG therapy is effective for improvement of QOL in some patients with metastatic malignant pheochromocytoma. Radioimmuno-therapy using anti-CD20 has been used successfully in clinical application in patients with malignant B cell lymphoma.     

Effective strategies for forensic analysis in the mitochondrial DNA coding region

Recently, it has been recognized that accessing information in the mtDNA coding region can provide additional forensic discrimination with respect to the standard typing of the D-loop region, augmenting the sometimes rather limited forensic power of mtDNA testing. Here, we discuss considerations relating to maximally effective approaches for recovering additional discrimination in the coding region, bearing in mind that (1) DNA quality and quantity in typical mtDNA casework usually restrict the amount of additional sequence that can be obtained, and (2) the need for additional discrimination primarily arises when common HV1/HV2 types are encountered. Most investigators have sought additional discrimination by sequencing short segments of coding region that are thought to be particularly variable. Unfortunately, efforts in this regard have generally failed to appreciate that most variation in the coding region is redundant with information already present in HV1/HV2 and have therefore overvalued the potential of this approach for providing additional discrimination. An alternative single nucleotide polymorphism-based approach [Int J Legal Med 118:137-146, 2004] has been to identify specific bases that provide resolution in specific common HV1/HV2 types (and related sequences). We investigate several highly relevant data sets wherein the latter approach performs appreciably better than sequencing selected short portions of the coding region. This is true even when only synonymous variation is targeted to minimize the potential for problems arising from discovery of mutations that have reportedly been related to disease.     

Sequence analysis of mitochondrial DNA hypervariable region I in Thai individuals

Nucleotide sequence analysis of hypervariable region I (HVRI) in human mitochondrial DNA (mtDNA) was investigated in 100 unrelated Thai individuals. A total of 85 variable sites and 423 base substitutions, which consisted of 390 nucleotide transitions and 33 nucleotide transversions were found. The following nucleotide substitutions were found: 48% at 16,223, 31% at 16,304, 30% at 16,332, and 26% at 16,129, respectively. Transition from T to C (43.7%) was the most frequent substitution. The nucleotide insertions were found at two sites with T at position 16,188 and C at position 16,194. Eighty-two haplotypes were investigated of which 72 haplotypes were unique. The most frequent haplotypes (16,108T-16,129A-16,162G-16,172C-16,304C and 16,260T-16,298C-16,355T-16,362C) were observed. From position 16,180 to 16,193, thirteen patterns of polycytosine or C-stretch were observed, whereas 68 Thai individuals were found to be similar to the references. The genetic diversity, random match probability, and discrimination power were estimated to be 0.9943, 0.0156, and 0.9844, respectively.     

Automated DNA extraction of single dog hairs without roots for mitochondrial DNA analysis

Dogs are intensely integrated in human social life and their shed hairs can play a major role in forensic investigations. The overall aim of this study was to validate a semi-automated extraction method for mitochondrial DNA analysis of telogenic dog hairs. Extracted DNA was amplified with a 95% success rate from 43 samples using two new experimental designs in which the mitochondrial control region was amplified as a single large (± 1260 bp) amplicon or as two individual amplicons (HV1 and HV2; ± 650 and 350 bp) with tailed-primers. The results prove that the extraction of dog hair mitochondrial DNA can easily be automated to provide sufficient DNA yield for the amplification of a forensically useful long mitochondrial DNA fragment or alternatively two short fragments with minimal loss of sequence in case of degraded samples.     

妇产科学进展与发展设想

目的 综述国内外妇产科学的最新进展及研究动态,为研究我军今后妇产科相关技术的发展方向提供依据.方法 分别通过PubMed及CHKD检索近10年来妇产科的国内外文献,就妇产科领域的新进展、新技术进行分析、归纳及总结.结果 近10年来,妇产科领域取得的进展有:微创概念及微创手术迅速发展,内镜手术、介入治疗、经阴道及小切口等微创化手术的术式得到普及与发展,随着微创技术的发展,恶性肿瘤的早期诊断技术也随之提高;辅助生殖技术逐渐成熟,在此基础上提出了卵巢移植观点,卵巢移植技术为年轻恶性肿瘤患者提供了妊娠的希望;产前诊断技术已发展到种植前遗传学诊断水平,可以在妊娠更早的阶段对遗传缺陷疾病进行筛查,及时发现和处理存在遗传缺陷的胎儿.结论 "十二五"期间,我军妇产科学的发展应紧跟最新前沿动态,发展最新技术,并将这些技术向卫勤保障及战时应用转化.     

Evaluation of mitochondrial DNA coding region assays for increased discrimination in forensic analysis

There is an increasing trend to use mitochondrial DNA (mtDNA) analysis in criminal investigations where only limited amounts of DNA are available. However, analysis of the mtDNA control region has the drawback of low discrimination power, due to the lack of recombination that results from uniparental (maternal) inheritance. As a strategy to increase discrimination, a number of typing assays detecting variation in the mitochondrial coding region have been developed. In this study, several of these assays are evaluated for their discriminatory capacity using data obtained from 495 complete Caucasian mtDNA sequences. In order to add a local geographic perspective to this evaluation, we have also sequenced and analysed the entire mtDNA from 20 individuals of Swedish origin. We find that the coding region assays are very useful for resolving sequences with identical HVI/HVII regions. The best-performing coding region assay was able to discriminate 46% of the resolvable sequences, compared to 20–30% for the other coding region assays we evaluated.     

Nucleotide sequence analysis of the hypervariable region III of mitochondrial DNA in Thais

This study analyzed the nucleotide sequences of the hypervariable region III (HVRIII) of mitochondrial DNA in Thai individuals. Buccal swab samples were randomly obtained from 100 healthy, unrelated, adult (18–60 years old), volunteer donors living in Thailand. Eighteen different haplotypes were found, of which 11 haplotypes were unique. The most frequent haplotypes observed were 522D-523D. Nucleotide transition from Thymine (T) to Cytosine (C) at position 489 (43%) was the most frequent substitution. Nucleotide transversions were also observed at position 433 (Adenine (A) to C, 1%) and position 499 (Guanine (G) to C, 1%). Fifty-three samples presented nucleotide insertion and deletion of C and A (CA) at position 514–523. Insertion of 1AC (3%) and 2AC (2%) were observed. Deletion of 1CA (53%) and 2CA (2%) at position 514–523 were revealed. The deletion of T at position 459 was observed. The haplotype diversity, random match probability, and discrimination power were calculated to be 0.7770, 0.2308, and 0.7692, respectively.     

血液代用品的研究现状和进展

综述了携氧扩充剂的种类，副作用及研究进展，血液代用品包括化学合成的化合物和天然及用重组技术得到的血红蛋白产品，如无基质血红蛋白，交联血红蛋白和脂质体包封血红蛋白。血液代用品必须是供给组织氧和提供维持生命的液体。     

介入分子影像学研究进展

近年来分子影像学迅速发展,为分子生物学、临床靶向治疗学等相关领域研究提供了有力的活体内监测手段.但目前多种分子影像技术在临床应用均存在一定的局限性,其对大动物乃至人的研究工作受到极大限制,使得分子影像学仍处于小动物基础成像或临床前研究阶段.介入分子影像学的出现,为解决这一系列问题提供了新思路,通过优化分子探针导入方式、改良现有分子成像技术装置等,使分子影像学从小动物基础研究发展为大动物研究和临床应用研究成为可能,并最终成为临床转化的重要桥梁.同时,介入分子影像学融合了分子影像诊断学与临床靶向治疗学,这无疑将成为推动临床靶向治疗及个体化治疗的重要力量,对未来临床诊治工作产生又一革命性影响,也是未来介入放射学发展的重要方向.     

Recent progress in advanced cardiac life support

The revised guidelines for advanced cardiac life support (ACLS) from the American Heart Association are anticipated in the fall of 2000. Although dramatic changes in the approach to adult basic and ACLS are not anticipated, several controversies and new drugs on the horizon may radically change our approach to emergent cardiac resuscitation. This article features some of the evolving thinking on the emergent treatment of the adult with ventricular fibrillation or ventricular tachycardia, the critical rhythms seen in most cases of acute cardiac distress. Approaches to airway therapy drug administration and new agents also are described.     

康复医学新进展与发展设想

目的根据国内外军队康复与理疗医学专业研究现状和发展动态,提出未来我军康复与理疗专业的发展方向。方法采用文献检索方法,复习近10年来国内外该领域的主要文献,明确该学科的现状和发展趋势。同时发放军队康复理疗科问卷调查表,收集来自基层医院的信息,进行综合分析。结果十一五以来,我军康复与理疗学专业有了长足进步,涌现出一批学科发展明确,技术手段先进,医疗服务优良的学科和科室。结论学科发展的总体构想是进一步探索康复医学新理论、新技术,使其进一步适应军队卫勤保障需求,并向现代康复与理疗医学方向发展。     

胚胎皮肤无瘢痕愈合机制研究进展

创伤后的愈合方式是创伤愈合过程中的重要环节,胎儿皮肤创伤后的无瘢痕愈合是人们追求的理想愈合模式。胎儿的无瘢痕愈合与其生长环境、创伤的炎症反应、纤维母细胞及细胞外基质、以及众多细胞因子的调节等有密切的关系。笔者对近年来关于胚胎皮肤无瘢痕愈合的可能相关机制研究进行综述如下。     

AQME: A forensic mitochondrial DNA analysis tool for next-generation sequencing data

The feasibility of generating mitochondrial DNA (mtDNA) data has expanded considerably with the advent of next-generation sequencing (NGS), specifically in the generation of entire mtDNA genome (mitogenome) sequences. However, the analysis of these data has emerged as the greatest challenge to implementation in forensics. To address this need, a custom toolkit for use in the CLC Genomics Workbench (QIAGEN, Hilden, Germany) was developed through a collaborative effort between the Armed Forces Medical Examiner System − Armed Forces DNA Identification Laboratory (AFMES-AFDIL) and QIAGEN Bioinformatics. The AFDIL-QIAGEN mtDNA Expert, or AQME, generates an editable mtDNA profile that employs forensic conventions and includes the interpretation range required for mtDNA data reporting. AQME also integrates an mtDNA haplogroup estimate into the analysis workflow, which provides the analyst with phylogenetic nomenclature guidance and a profile quality check without the use of an external tool. Supplemental AQME outputs such as nucleotide-per-position metrics, configurable export files, and an audit trail are produced to assist the analyst during review. AQME is applied to standard CLC outputs and thus can be incorporated into any mtDNA bioinformatics pipeline within CLC regardless of sample type, library preparation or NGS platform.An evaluation of AQME was performed to demonstrate its functionality and reliability for the analysis of mitogenome NGS data. The study analyzed Illumina mitogenome data from 21 samples (including associated controls) of varying quality and sample preparations with the AQME toolkit. A total of 211 tool edits were automatically applied to 130 of the 698 total variants reported in an effort to adhere to forensic nomenclature. Although additional manual edits were required for three samples, supplemental tools such as mtDNA haplogroup estimation assisted in identifying and guiding these necessary modifications to the AQME-generated profile. Along with profile generation, AQME reported accurate haplogroups for 18 of the 19 samples analyzed. The single errant haplogroup assignment, although phylogenetically close, identified a bug that only affects partial mitogenome data. Future adjustments to AQME’s haplogrouping tool will address this bug as well as enhance the overall scoring strategy to better refine and automate haplogroup assignments. As NGS enables broader use of the mtDNA locus in forensics, the availability of AQME and other forensic-focused mtDNA analysis tools will ease the transition and further support mitogenome analysis within routine casework. Toward this end, the AFMES-AFDIL has utilized the AQME toolbox in conjunction with the CLC Genomics Workbench to successfully validate and implement two NGS mitogenome methods.     

Rapid screening for Native American mitochondrial and Y-chromosome haplogroups detection in routine DNA analysis

Aiming to detect individuals of Native American maternal or paternal ancestry a rapid screening approach has been developed. Its strategy was based on SNP typing by Real Time PCR (rt-PCR) followed by High Resolution Melting analysis (HRM). After extraction, DNA was quantitated by rt-PCR using commercial kits; samples were then submitted to two multiplex reactions in order to determine the major Native American mtDNA and Y-chromosome haplogroups by HRM. One cocktail included primers flanking nucleotide substitutions that define mtDNA haplogroup C and sub-haplogroups A2, B2, and D1. The other included primers flanking Y-SNPs M3, M269 and U179 that allowed discriminating Q and non-Q haplogroups. In all cases amplicons were <125 nucleotides long in order to increase the peak resolution. The accuracy of the results obtained was established by means of sequencing analysis of the amplicons. The new working-flow here proposed facilitates and speeds-up the screening process that may preclude a detailed sequencing analysis of particular samples, or for further molecular epidemiological investigations in which continental origin influences might be relevant.     

Recent progress in fluorine-18 labelled peptide radiopharmaceuticals

The application of biologically active peptides labelled with positron-emitting nuclides has emerged as a useful and interesting field in nuclear medicine. Small synthetic receptor-binding peptides are currently the preferred agents over proteins and antibodies for diagnostic imaging of various tumours. Due to the smaller size of peptides, both higher target-to-background ratios and rapid blood clearance can often be achieved with radiolabelled peptides. Hence, short-lived positron emission tomography (PET) isotopes are potential candidates for labelling peptides. Among a number of positron-emitting nuclides, fluorine-18 appears to be the best candidate for labelling bioactive peptides by virtue of its favourable physical and nuclear characteristics. The major disadvantage of labelling peptides with ¹⁸F is the laborious and time-consuming preparation of the ¹⁸F labelling agents. In recent years, various techniques have been developed which allow efficient labelling of peptides with ¹⁸F without affecting their receptor-binding properties. Moreover, the development of a variety of prosthetic groups has facilitated the efficient and site-specific labelling of peptides with ¹⁸F. The ¹⁸F-labelled peptides hold enormous clinical potential owing to their ability to quantitatively detect and characterise a wide variety of human diseases when using PET. Recently, a number of ¹⁸F-labelled bioactive peptides have shown great promise as diagnostic imaging agents. This review presents the recent developments in ¹⁸F-labelled biologically active peptides used in PET.     

Role of mitochondrial DNA in radiation exposure

PURPOSE: To evaluate the role of mitochondrial DNA (mtDNA) following exposure to ionizing irradiation. MATERIALS AND METHODS: We examined two human osteosarcoma cell lines either lacking mtDNA (143B.rho(0)206; rho0 cells) or having normal mtDNA (143B.TK-; rho+ cells). Cell survival curves were generated by using colony formation and micronucleus assay. The delay in population doubling time after irradiation was evaluated with dye exclusion tests. RESULTS: No significant difference was seen between rho+ and rho0 cell lines in colony formation assay. In micronucleus assay, rho0 cells showed a significantly lower rate of micronucleus formation. The ratios of binucleated cells with micronuclei were 0.49 for rho+ cells and 0.25 for rho0 cells (p=0.005). In the dye exclusion test, rho0 cells revealed a delay of about 1.6 times in population doubling time compared with the control after 5 Gy of irradiation, similar to the 1.7 times of rho+ cells. CONCLUSION: In the human osteosarcoma cell line 143B.TK-, mtDNA does not influence clonogenic survival and delay of population doubling time after irradiation. However, the difference in micronucleus formation shows that mtDNA influences DNA damage after radiation exposure.