Spinal cord sensory systems after neonatal capsaicin

Animals with a severe reduction in the number of afferent C-fibres as a consequence of neonatal administration of capsaicin, exhibit a number of neurological and behavioral deficits including increased nociceptive thresholds, altered somato-visceral and viscero-visceral reflexes, depressed cardiovascular and respiratory reflexes and changes in the organisation of spinal cord sensory systems. The reduction in the number of C-fibres produced by neonatal capsaicin does not cause a decrease of similar magnitude in the number of dorsal horn cells driven by the surviving C-fibres. Twenty-two per cent of dorsal horn neurones in capsaicin treated animals respond to electrical stimulation of the surviving afferent C-fibres: a reduction of only 50% from control values. Inhibitory controls on afferent C-fibre evoked responses of dorsal horn neurones are weaker in capsaicin treated rate than in control animals. The cutaneous receptive fields of some dorsal horn neurones can increase in size following stimulation of afferent C-fibres. Tonic descending inhibition on C-fibre evoked responses of dorsal horn neurones is reduced in capsaicin treated rats: fewer neurones show tonic descending inhibition in these animals and those that do are subjected to less powerful inhibitions than similar neurones from control animals. However, some central inhibitory mechanism are unchanged after neonatal capsaicin treatment, specially those that do not involve afferent C-fibres. We suggest that the nervous system develops central inhibition in response to and directed towards the excitations mediated by its afferent drives. Therefore reduced central inhibition in response to a decreased number of afferent C-fibres can compensate for the lost capacity in the signalling of peripheral noxious events.     

蛛网膜下腔移植脐血干细胞治疗一氧化碳中毒后迟发性脑病的临床研究

目的观察蛛网膜下腔途径移植脐血干细胞治疗一氧化碳中毒后迟发性脑病（DEACMP）的安全性及治疗效果。方法26例DEACMP患者,按治疗方式分两组,其中12例经脐血干细胞移植治疗为治疗组,14例经高压氧治疗为对照组,对每例患者在治疗前和治疗后进行MMSE评分、ADL评分及颅脑CT检查。所有患者连续随访1年,观察预后及有无不良反应。采用两样本t检验、方差分析及卡方检验进行统计学分析。结果治疗组患者在干细胞移植术中均无特殊不适感,术中及术后24h内生命体征平稳,无发热,无穿刺点感染、出血等,术后门诊随访,均未出现移植相关并发症。1年后治疗组患者MMSE评分为25．08±4．10,高于对照组11．7±10．33（t=4．199,P=0．0003）;治疗组ADL评分为93．33±13．37,高于对照组41．07±38．39（t=4．478,P=0．0002）,差异均有统计学意义。治疗组CT检查结果好转率为83．33％,对照组好转率为30．00％两组比较差异有统计学意义（Х^2=4．402,P=0．0359）;治疗组临床痊愈率为50．00％,对照组为7．69％,差异有统计学意义（Х^2=4．055,P=0．0442）;治疗组临床有效率为100．00％,对照组为50．00％差异有统计学意义（Х^2=5．866,P=0．0154）。结论脐血干细胞移植治疗DEACMP疗效优于高压氧治疗,是一种安全有效的生物治疗方法,应得到重视和深入的研究。     

Optimal umbilical cord blood collection,processing and cryopreservation methods for sustained public cord blood banking

Background aimsUmbilical cord blood is an established source of stem cells in patients with hematologic malignancies who do not have HLA-compatible matched related or unrelated donors. The success of an umbilical cord blood transplant depends on the dose of total nucleated and CD34+ cells infused. Therefore, collecting, banking and listing high-quality cord blood units with high total nucleated and CD34+ cell dose are essential.MethodsHere the authors describe their cord blood bank's novel collection technique, which involves both in utero and ex utero collection of a single cord blood unit. The authors also evaluated maternal, neonatal and collection parameters that may impact the cell dose.ResultsMaternal gestational age and race, and neonatal weight and sex correlated with the total nucleated cell dose.ConclusionsThe optimized collection of umbilical cord blood is critical for its use as a source of stem cells for transplantation.     

New allergies after cord blood transplantation

Background aimsUmbilical cord blood transplantation (CBT) is an effective treatment for benign and malignant diseases. Late effects of CBT are not well described in the literature. In the present study, we present our experience of new-onset allergies in long-term survivors after CBT.MethodsAfter an initial patient had a severe peanut allergic reaction after CBT, all CBT patients were prospectively followed for new allergy development. Fifty patients received CBT between March 2006 and June 2011.ResultsThe median follow-up after CBT was 447 days (range, 12–2022). At the time of analysis, 30 patients were alive, with 3-year survival of 55.5%; median follow-up of surviving patients was 910 days (range, 68–2022). The allergic syndrome developed in five patients, with the cumulative incidence of new allergies at 2 years of 18.4% (95% confidence interval, 10.8–26). The median time to onset of new allergy after transplantation was 298 days (range, 250–809).ConclusionsAllergy development has been linked to a delayed maturation of the immune system in several studies. We present the first case series of patients who had new allergies after CBT. Further study of this novel complication as well as counseling of patients after CBT would be important.     

Inter- and intra-individual variation in plasma and red blood cell vitamin E after supplementation

To establish the range of individual blood responses to supplemental vitamin E, 30 healthy subjects ingested 75 mg of deuterium-labelled α-tocopherol with a standard breakfast. Blood was collected at 6, 9, 12, 27 and 51 h post ingestion and deuterated (d₆) and non-deuterated (d₀) α-tocopherol concentrations were determined in plasma and red blood cells (RBC) by GC-MS. To examine intra-individual responses, 6 of these subjects were re-examined at 6-month intervals over a 30-month period. Post ingestion, the amount of d₆-α-tocopherol in blood increased rapidly with time with maximal concentrations seen at 12 h (plasma) and 27 h (RBC) in most subjects. At these times, d₆-α-tocopherol concentration ranged from 0.3–12.4 μmol/l in plasma and 0.6–4.09 μmol/l packed cell in RBC. Area under the curve calculations indicated inter-individual differences of α-tocopherol uptake to be 40-fold for plasma (12.9–493.3 μmol h/l) and 6-fold for RBC (24.4–146.1 μmol h/l packed RBC). Intra-individual variation in α-tocopherol uptake was small in comparison and remained relatively constant over the 30-month period. We conclude that vitamin E uptake varies widely in the normal population, although it is comparatively stable for an individual over time. These differences likely arise from variations in the regulation of vitamin E uptake and metabolism between subjects. Factors regulating this process must be better understood before the optimal intake of vitamin E can be ascertained.     

Inter- and intra-individual variation in plasma and red blood cell vitamin E after supplementation

To establish the range of individual blood responses to supplemental vitamin E, 30 healthy subjects ingested 75 mg of deuterium-labelled alpha-tocopherol with a standard breakfast. Blood was collected at 6, 9, 12, 27 and 51 h post ingestion and deuterated (d6) and non-deuterated (do) alpha-tocopherol concentrations were determined in plasma and red blood cells (RBC) by GC-MS. To examine intra-individual responses, 6 of these subjects were re-examined at 6-month intervals over a 30-month period. Post ingestion, the amount of d6-alpha-tocopherol in blood increased rapidly with time with maximal concentrations seen at 12 h (plasma) and 27 h (RBC) in most subjects. At these times, d6-alpha-tocopherol concentration ranged from 0.3-12.4 micromol/l in plasma and 0.6-4.09 micromol/l packed cell in RBC. Area under the curve calculations indicated inter-individual differences of alpha-tocopherol uptake to be 40-fold for plasma (12.9-493.3 micromol h/l) and 6-fold for RBC (24.4-146.1 micromol h/l packed RBC). Intra-individual variation in alpha-tocopherol uptake was small in comparison and remained relatively constant over the 30-month period. We conclude that vitamin E uptake varies widely in the normal population, although it is comparatively stable for an individual over time. These differences likely arise from variations in the regulation of vitamin E uptake and metabolism between subjects. Factors regulating this process must be better understood before the optimal intake of vitamin E can be ascertained.     

Neonatal plasma lipids as measured in cord blood

The status of cord blood screening of plasma lipids is reviewed, emphasizing the problems associated with the diagnosis of familial hyperlipoproteinemia (HLP), particularly type II (hyperbetalipoproteinemia), in the neonate. For 2937 neonates in this study the mean ± standard deviation (SD) of plasma cholesterol was 70.3 ± 16.9 mg/dl. The mean triglyceride level of 1805 neonates was 39.6 ± 19.3 mg/dl. The mean level of β-cholesterol in 240 neonates was 34.3 ± 10.9 mg/dl and the correlation between total and β-cholesterol was high (r=0.89). By convention, the upper limit of normal was defined as the mean + 2 SD, and these levels for total cholesterol, triglyceride and β-cholesterol were 100, 80 and 55 mg/dl respectively. Only by careful follow-up of hyperlipidemic neonates can the incidence of HLP and the worth of early diagnosis be assessed.     

Modification of blood plasma crystallogenesis with nitrogen oxide processing

The aim of this work is to estimate crystallogenic properties of human blood serum processed by gas nitric oxide. Blood samples of ten healthy patients were processed by cold plasma with NO (800 μg/L). The true and initiated (by 0.9% sodium chloride solution) crystallogenesis of non- and NO-processed blood serum was estimated. It was stated, that as a result of gas nitric oxide processing crystallogenic properties of blood serum undergo a series of specific and nonspecific transformations, the most obvious one is the formation of a raised band in the marginal zone of the facia.     

Immune reconstitution after unrelated cord blood transplantation

Over the past years unrelated cord blood transplant (UCBT) has emerged as an effective alternative to unrelated donor blood and marrow transplantation. However, despite several advantages, its success is limited by the high incidence of opportunistic infections (OI), most of which are viral. Infection-related mortality is the primary cause of death after UCBT with most deaths occurring in the first 3-6 months post transplant. For several months, until recovery of the thymus is restored to support de novo T cell generation, protective antiviral immunity depends on the activity of post-thymic T cells infused within the cord blood (CB) grafts. However, almost all CB T cells are antigen inexperienced (na?ve) lymphocytes that have been functionally altered by placental factors to protect pregnancy. CB T cells need to undergo in vivo priming, Th1/Tc1 maturation, and peripheral expansion before they can afford immunologic protection. This article provides an overview of what is currently known regarding the reconstitution of adaptive immunity following UCBT including our own data from prospective analyses of pediatric cohorts. Remarkable immunophenotypic changes are notable already in the first 2-3 weeks post-UCBT. These changes result from apparent 'homeostatic' peripheral T cell expansion in the lymphopenic environment. While we can identify patient- and graft-specific predictive factors, the concordant emergence of T cell subsets displaying the phenotype of Th1/Tc1 cytotoxic effector cells can be statistically linked to those UCBT recipients who will subsequently develop viral and other opportunistic infections. Antigen presenting dendritic cell reconstitution may also reflect alterations in immunocompetence due to OI and/or GVHD.     

脐血与老人外周血中TIMP2蛋白检测的影响因素及意义

目的探讨不同血浆处理方式对脐血和老人外周血血浆中TIMP2蛋白检测的影响,建立人血浆中TIMP2蛋白检测方法的同时为联合干细胞延缓衰老提供理论基础。 方法采集了15份脐带血和15份老人外周血,利用不同的离心力、不同的储存温度和不同的检测试剂盒对血浆中TIMP2蛋白最终检测含量的影响。多组均数差异比较采用单因素方差分析,两组比较采用独立样本t检验。 结果1?000×g、1?500×g和2?000×g离心力组组间TIMP2蛋白含量差异无统计学意义（P?> 0.05）。4 ℃、-20?℃、-80?℃保存组,保存一周或者两周TIMP2蛋白含量差异均无统计学意义（P?> 0.05）。利用R&D Systems、Abnova和Abcam检测试剂盒均不对全血中TIMP2蛋白最终检测量产生显著影响（P?>?0.05）。脐带血中的TIMP2蛋白含量（147.00±15.92）?pg/ml高于老人外周血中TIMP2蛋白含量（79.56±10.80）?pg/?ml,差异具有统计学意义（P?< 0.01）。 结论不同离心力、血浆储存温度及检测试剂盒间均不影响脐血和老人外周血中TIMP2蛋白最终的检测含量,且脐血血浆中TIMP2蛋白含量高于老人外周血。显示TIMP2蛋白可能与衰老存在直接的相关性,具有与干细胞联合治疗并延缓衰老的潜在价值。     

Expanded umbilical cord blood T cells used as donor lymphocyte infusions after umbilical cord blood transplantation

BackgroundUmbilical cord blood (UCB) is an alternative graft source for hematopoietic stem cell transplantation and has been shown to give results comparable to transplantation with other stem cell sources. Donor lymphocyte infusion (DLI) is an effective treatment for relapsed malignancies after hematopoietic stem cell transplantation. However, DLI is not available after UCB transplantation.MethodsIn this study, in vitro–cultured T cells from the UCB graft were explored as an alternative to conventional DLI. The main aim was to study the safety of the cultured UCB T cells used as DLI because such cell preparations have not been used in this context previously. We also assessed potential benefits of the treatment.ResultsThe cultured UCB T cells (UCB DLI) were given to 4 patients with mixed chimerism (n = 2), minimal residual disease (n = 1) and graft failure (n = 1). No adverse reactions were seen at transfusion. Three of the patients did not show any signs of graft-versus-host disease (GVHD) after UCB DLI, but GVHD could not be excluded in the last patient. In the patient with minimal residual disease treated with UCB DLI, the malignant cell clone was detectable shortly before infusion but undetectable at treatment and for 3 months after infusion. In 1 patient with mixed chimerism, the percentage of recipient cells decreased in temporal association with UCB DLI treatment.ConclusionsWe saw no certain adverse effects of treatment with UCB DLI. Events that could indicate possible benefits were seen but with no certain causal association with the treatment.     

人脐血血浆外泌体提取方法的比较北大核心CSCD

为探寻高效且稳定的提取人脐血血浆外泌体的方法,利用超高速离心法、蔗糖垫密度梯度离心法、改良超速离心法和聚乙二醇(polyethylene glycol, PEG)沉淀法提取人脐血血浆外泌体,并比较4种方法的优劣。利用透射电镜、动态光散射技术观察外泌体的形态、结构及大小;聚氰基丙烯酸正丁酯(bicinchoninic acid, BCA)法测定外泌体蛋白总量;Western blotting检测外泌体表面标志蛋白CD63、HSP70以及外泌体阴性蛋白GM130 (高尔基标志蛋白)的表达。结果表明,与提取外泌体的“金标准”,即超高速离心法相比,蔗糖垫密度梯度离心法稳定性好,获取的外泌体粒径较均一,但操作较复杂,耗时长;改良超速离心法操作较简单,纯度较高;PEG沉淀法提取的外泌体蛋白量最高,操作时间最短,但杂质较多。结果表明,4种方法均能从人脐血血浆中获取外泌体,但在操作时间、纯度、提取量等方面存在一定差异。因此,应根据实验目的和具体要求选择合适的提取人脐血血浆外泌体的方法。     

Cancer associated proteins in blood plasma: Determining normal variation

Protein biomarkers have the potential to improve diagnosis, stratification of patients into treatment cohorts, follow disease progression and treatment response. One distinct group of potential biomarkers comprises proteins which have been linked to cancer, known as cancer associated proteins (CAPs). We determined the normal variation of 86 CAPs in 72 individual plasma samples collected from ten individuals using SRM mass spectrometry. Samples were collected weekly during 5 weeks from ten volunteers and over one day at nine fixed time points from three volunteers. We determined the degree of the normal variation depending on interpersonal variation, variation due to time of day, and variation over weeks and observed that the variation dependent on the time of day appeared to be the most important. Subdivision of the proteins resulted in two predominant protein groups containing 21 proteins with relatively high variation in all three factors (day, week and individual), and 22 proteins with relatively low variation in all factors. We present a strategy for prioritizing biomarker candidates for future studies based on stratification over their normal variation and have made all data publicly available. Our findings can be used to improve selection of biomarker candidates in future studies and to determine which proteins are most suitable depending on study design.     

Application of the accurate mass and time tag approach in studies of the human blood lipidome

We report a preliminary demonstration of the accurate mass and time (AMT) tag approach for lipidomics. Initial data-dependent LC-MS/MS analyses of human plasma, erythrocyte, and lymphocyte lipids were performed in order to identify lipid molecular species in conjunction with complementary accurate mass and isotopic distribution information. Identified lipids were used to populate initial lipid AMT tag databases containing 250 and 45 entries for those species detected in positive and negative electrospray ionization (ESI) modes, respectively. The positive ESI database was then utilized to identify human plasma, erythrocyte, and lymphocyte lipids in high-throughput LC-MS analyses based on the AMT tag approach. We were able to define the lipid profiles of human plasma, erythrocytes, and lymphocytes based on qualitative and quantitative differences in lipid abundance.     

Chemotaxonomic characterisation of the thaumarchaeal lipidome

Thaumarchaeota are globally distributed and abundant microorganisms occurring in diverse habitats and thus represent a major source of archaeal lipids. The scope of lipids as taxonomic markers in microbial ecological studies is limited by the scarcity of comparative data on the membrane lipid composition of cultivated representatives, including the phylum Thaumarchaeota. Here, we comprehensively describe the core and intact polar lipid (IPL) inventory of ten ammonia‐oxidising thaumarchaeal cultures representing all four characterized phylogenetic clades. IPLs of these thaumarchaeal strains are generally similar and consist of membrane‐spanning, glycerol dibiphytanyl glycerol tetraethers with monoglycosyl, diglycosyl, phosphohexose and hexose‐phosphohexose headgroups. However, the relative abundances of these IPLs and their core lipid compositions differ systematically between the phylogenetic subgroups, indicating high potential for chemotaxonomic distinction of thaumarchaeal clades. Comparative lipidomic analyses of 19 euryarchaeal and crenarchaeal strains suggested that the lipid methoxy archaeol is synthesized exclusively by Thaumarchaeota and may thus represent a diagnostic lipid biomarker for this phylum. The unprecedented diversity of the thaumarchaeal lipidome with 118 different lipids suggests that membrane lipid composition and adaptation mechanisms in Thaumarchaeota are more complex than previously thought and include unique lipids with as yet unresolved properties.