人外周血树突状细胞的分离与鉴定

目的:体外分离培养并鉴定人外周血树突状细胞,并观察其抗原呈递功能。方法:实验于2005-05/2006-11在南方医科大学南方医院肿瘤中心生物治疗实验室完成。从人类白细胞抗原A2表达阳性的健康人外周血中分离获得单个核细胞。培养5h后洗涤贴壁细胞,加入含有10%人AB血清的RPMI1640培养基,及重组人粒细胞-巨噬细胞集落刺激因子和重组人白细胞介素4,于培养的第1,3,6天对树突状细胞的形态、表型进行分析,并定期检测树突状细胞的纯度与得率。抽取与以上树突状细胞不同来源的其他健康人外周血。经淋巴细胞分离液分离后,获取非贴壁细胞,用含10%人AB血清的1640培养基重悬,加入白细胞介素2继续孵育6d,作为同种异体T淋巴细胞。将树突状细胞分为两组,一组按常规方法培养6d,另一组在培养至第5天时加入黑色素瘤抗原基因A3编码的多肽继续培养24h。在经紫外线处理后的96孔板中,分别加入树突状细胞悬液1×104,5×103,2×103,1×103细胞/每孔,以自身T淋巴细胞作为对照,每孔设3个复孔,分别加入1×105淋巴细胞/每孔。评价树突状细胞刺激T淋巴细胞增殖的能力。结果:①单个核细胞体外培养至第6天,可获得大量、90.81%高纯度的树突状细胞,能够较高地表达21.8?1a、99.0%HLA-DR、63.4?80、18.9?83和80.6?86。②将诱导培养6d获得的两组树突状细胞作为刺激细胞,以不同的浓度与同种异体淋巴细胞混合,均可产生增殖反应;经过黑色素瘤抗原基因A3编码的多肽处理的各种比例的树突状细胞,较相应未经黑色素瘤抗原基因A3编码的多肽处理的树突状细胞激发淋巴细胞增殖的能力明显增强,浓度相对较高的树突状细胞刺激效果最明显,能够强烈地激发同种混合淋巴细胞增殖。结论:得到了一群较高程度表达CD83、CD86和HLA-DR分子、体外可强烈激发同种异体T淋巴细胞增殖的树突状细胞群。     

Do migrainous headaches become more consistently lateralized?

Analysis of information collected from 468 patients with common or classical migraine does not suggest that those with a longer history are more likely to have headaches fixed to a single side of the head. This finding does not support the notion that successive unilateral headache attacks on one side cause damage that predisposes the patient to further attacks on the same side.     

Role of prostaglandin E in the biphasic fever response to endotoxin

Biphasic fevers were induced in sheep with intravascular infusions or injections of 4-10 μg (80-200 ng/kg) of endotoxin, whereas monophasic fevers were obtained with doses of 1-2/μg (20-40 ng/kg). A marked increase in arterial blood pressure invariably accompanied the onset of fever; the latency of responses to the higher and lower doses of endotoxins averaged 26 min and 42 min, respectively. Prostaglandin (PG) assays of plasma from the carotid artery and jugular vein during fever episodes revealed a surge of PGE and PGF coincident with the pressor response and the first phase of fever, but PG were not detected in plasma samples taken throughout the second phase of fever. PG measurements of arterial and venous plasma collected at a distal site (hind limb) showed a similar surge of PGE and PGF in association with the early fever response, indicating that intravascular PG synthesis and release represents a generalized systemic response to circulating endotoxin. Carotid arterial infusions of PGE(2) produced immediate monophasic fevers and pressor responses, whereas PGD(2) infusions produced an immediate pressor effect but no fever. Infusions of PGF(2α) or prostacyclin, however, evoked neither fever nor pressor effects. Intracarotid infusions of leukocyte pyrogen (LP) caused monophasic fevers with latent periods of 15-20 min but pressor responses were not seen and neither PGE nor PGF were detected in plasma samples from the carotid artery or jugular vein before or during fever. Indomethacin, a potent inhibitor of arachidonic acid metabolism, blocked fever responses to endotoxin and to LP. These findings implicate PGE as the mediator of the early phase of endotoxin fever and imply a role for another pyrogenic metabolite ofarachidonic acid in the mediation of the second phase of fever, i.e., the phase associated with circulating LP. It is possible that both pyrogenic metabolites are generated within the vascular compartment, reaching thermoregulatory centers of the brain by transfer across the blood-brain interface.     

Inhibition of airway remodeling in IL-5-deficient mice

To determine the role of IL-5 in airway remodeling, IL-5-deficient and WT mice were sensitized to OVA and challenged by repetitive administration of OVA for 3 months. IL-5-deficient mice had significantly less peribronchial fibrosis (total lung collagen content, peribronchial collagens III and V) and significantly less peribronchial smooth muscle (thickness of peribronchial smooth muscle layer, alpha-smooth muscle actin immunostaining) compared with WT mice challenged with OVA. WT mice had a significant increase in the number of peribronchial cells staining positive for major basic protein and TGF-beta. In contrast, IL-5-deficient mice had a significant reduction in the number of peribronchial cells staining positive for major basic protein, which was paralleled by a similar reduction in the number of cells staining positive for TGF-beta, suggesting that eosinophils are a significant source of TGF-beta in the remodeled airway. OVA challenge induced significantly higher levels of airway epithelial alphaVbeta6 integrin expression, as well as significantly higher levels of bioactive lung TGF-beta in WT compared with IL-5-deficient mice. Increased airway epithelial expression of alphaVbeta6 integrin may contribute to the increased activation of latent TGF-beta. These results suggest an important role for IL-5, eosinophils, alphaVbeta6, and TGF-beta in airway remodeling.     

骨组织工程研究中移植免疫的若干问题

目的:探讨骨组织工程中移植免疫研究进展。资料来源:应用计算机检索美国NCBI Pubmed数据库1998-01/2006-01关于骨组织工程中移植免疫研究的文章,主要检索主题词包括“bone tissue engineering,scaffold,antigen,immunity”,语言设为英语;检索2002-01/2006-01中文期刊相关文章,检索词为“骨组织工程,移植免疫,天然生物支架”。资料选择:选取报道骨组织工程中移植免疫研究的原创实验性文章。纳入标准:①具有原创性,论点论据可靠的实验文章。②观点明确,分析全面的文章。③文献主体内容与此课题联系紧密的文章。排除标准:实验设计不合理的文章及观点模糊的综述。质量评价主要考察资料的真实性、实施过程是否严密。资料提炼:共检索到关于骨组织工程中移植免疫研究的文献65篇,34篇文献符合纳入标准。资料综合:34篇文献分别说明了骨组织工程中支架材料、种子细胞及培养基的免疫原性,移植免疫的机制、降低移植免疫的对策等问题;分析了天然支架材料构象的保持、免疫原的消除、细胞黏附的关系等方面的影响因素。结果表明:①异种骨衍生支架材料的抗原性主要来自血管内皮组织及骨陷窝内的骨细胞;自体、同胎及同种异体或同种异基因细胞均可作为组织工程骨可靠的细胞来源,单纯异种成骨细胞不能作为组织工程骨的细胞来源。②异种细胞外基质材料移植的早期有明显的细胞反应,这些细胞主要为单核细胞,无特定的标志,未观察到浆细胞,Th1和Th2淋巴细胞介导宿主对异种细胞外基质材料免疫应答,引发宿主免疫排斥是暂时的或一过性的。③微囊具有较好的免疫隔离作用和生物相容性,免疫抑制剂可促进新骨形成,生物衍生骨经表面修饰后,支架材料表面性质、孔隙率及孔径均发生改变,有利于黏附、伸展及生长。结论:骨组织工程中天然支架材料、异体种子细胞及异种血清培养基均可带入异种抗原。Th1和Th2淋巴细胞介导宿主对异种细胞外基质材料免疫应答。引发宿主免疫排斥是暂时的或一过性的。免疫隔离、使用免疫抑制剂、改良细胞外基质材料均能减轻移植免疫。     

妊娠晚期胎儿眼外肌Pulley系统发育特征

目的:观察妊娠晚期胎儿眼外肌Pulley形态的异同点,分析人类早期眶内Pulley系统的发育特征,探讨眶内Pulley系统对人类眼球运动及双眼视发育的作用。方法:实验于2006-04/2007-04在天津市眼科研究所完成。孕龄为27周、30周、33周的新鲜胎儿尸头各1例(标本来源合法,并得到科研管理部门的批准)。取完整眼眶,整体固定,常规脱水,石蜡包埋,冠状位连续切片,Masson三染色显示胶原纤维。结果:人类早期眶内Pulley发育中LR-IO间的Pulley连接带首先被观察到有明显的带状发育;孕30周时外直肌Pulley环己较明显,外直肌与上直肌间连接带发育较好;内直肌周围己有胶原纤维环包绕;在孕33周胎儿眶内,内直肌纤维环发育较好,但内直肌与上直肌及内直肌与下直肌间的连接带仍未见明显发育。结论:人类的眼外肌周围结缔组织结构的发育具有时间与空间的顺序性。其发育顺序为颞下侧连接带开始,经外直肌Pulley环、外直肌-上直肌间连接带、上直肌Pulley环,逐渐向鼻侧发育。这提示内侧发达的纤维环及连接带可能是在生后随双眼视功能的发育而发育完善。     

The impact of the introduction of fidaxomicin on the management of <Emphasis Type="Italic">Clostridium difficile</Emphasis> infection in seven NHS secondary care hospitals in England: a series of local service evaluations

Clostridium difficile infection (CDI) is associated with high mortality. Reducing incidence is a priority for patients, clinicians, the National Health Service (NHS) and Public Health England alike. In June 2012, fidaxomicin (FDX) was launched for the treatment of adults with CDI. The objective of this evaluation was to collect robust real-world data to understand the effectiveness of FDX in routine practice. In seven hospitals introducing FDX between July 2012 and July 2013, data were collected retrospectively from medical records on CDI episodes occurring 12 months before/after the introduction of FDX. All hospitalised patients aged ≥18 years with primary CDI (diarrhoea with presence of toxin A/B without a previous CDI in the previous 3 months) were included. Recurrence was defined as in-patient diarrhoea re-emergence requiring treatment any time within 3 months after the first episode. Each hospital had a different protocol for the use of FDX. In hospitals A and B, where FDX was used first line for all primary and recurrent episodes, the recurrence rate reduced from 10.6 % to 3.1 % and from 16.3 % to 3.1 %, with a significant difference in 28-day mortality from 18.2 % to 3.1 % (p?<?0.05) and 17.3 % to 6.3 % (p?<?0.05) for hospitals A and B, respectively. In hospitals using FDX in selected patients only, the changes in recurrence rates and mortality were less marked. The pattern of adoption of FDX appears to affect its impact on CDI outcome, with maximum reduction in recurrence and all-cause mortality where it is used as first-line treatment.     

Migraine and menstruation: a pilot study

OBJECTIVE: To define the term "menstrual" migraine and to determine the prevalence of "menstrual" migraine in women attending the City of London Migraine Clinic. DESIGN: Women attending the clinic were asked to keep a record of their migraine attacks and menstrual periods for at least 3 complete menstrual cycles. RESULTS: Fifty-five women completed the study. "Menstrual" migraine was defined as "migraine attacks which occur regularly on or between days -2 to +3 of the menstrual cycle and at no other time". Using this criterion, 4 (7.2%) of the women in our population had "menstrual" migraine. All 4 women had migraine without aura. A further 19 (34.5%) had an increased number of attacks at the time of menstruation in addition to attacks at other times of the cycle. Eighteen (32.7%) had attacks occurring throughout the cycle but with no increase in number at the time of menstruation. Fourteen (25.5%) had no attacks within the defined period during the 3 cycles studied. DISCUSSION: A small percentage of women have attacks only occurring at the time of menstruation, which can be defined as true "menstrual" migraine. This group is most likely to respond to hormonal treatment. The group of 34.5% who have an increased number of attacks at the time of menstruation in addition to attacks at other times of the month could be defined as having "menstrually related" migraine and might well respond to hormonal therapy. The 32.7% who have attacks throughout the menstrual cycle without an increase at menstruation are unlikely to respond to hormonal therapy. The 25.5% who do not have attacks related to menstruation almost certainly will not respond to hormonal therapy.