血清中Zn,Cu,Mn,Cr和Se微量元素与口腔恶性肿瘤的关系

本研究测定与肿瘤关系密切的微量元素Ｚｎ、Ｃｕ、Ｍｎ、Ｃｒ和Ｓｅ在口腔颌面部正常者与良、恶性肿瘤患者血清中含量共９１例。结果表明微量元素Ｚｎ、Ｃｕ、Ｍｎ、Ｃｒ和Ｓｅ在口腔颌面部肿瘤与其它部位肿瘤一样有明显变化。用模式识别方法以微量元素作为指标，对良恶性肿瘤进行鉴别诊断，有一定的临床意义。     

Fertilization and early embryology: The effect of equilibration temperature and time on the outcome of ultrarapid freezing of 1-cell mouse embryos

Mouse 1-cell embryos were frozen ultrarapidly at a rate of 2500°C/minin solutions containing 0.25 M sucrose, 0.5% (w/v) bovine serumalbumin (BSA) and 3 or 4.5 M dimethyl sulphoxide (DMSO) or 3or 4.5 M 1, 2-propanediol (PROH) in HEPES-buffered modifiedEarle's medium. We investigated the effect of pre-freeze equilibrationfor 1, 3, 5 or 10 min at 22°C and for 1, 3, 5, 10, 15 or20 min at 4°C. After thawing in a 22°C water bath ata rate of 2500°C/min and dilution in 1 M sucrose in HEPES-bufferedmodified Earle's medium, embryos were cultured in vitro in bicarbonate-bufferedmodified Earle‘s medium with 0.5% (w/v) crystalline BSA.Embryo viability was expressed as the percentage of hatchingor hatched blastocysts resulting from the initial number offrozen-thawed 1-cell embryos. To determine the toxicity of thefreezing solutions, embryo viability was evaluated after equilibrationwithout freezing. Our results demonstrated that the concentration,the equilibration temperature and time are very important factorsin ultrarapid freezing of mouse 1-cell embryos. Optimal viabilitywas found when equilibration was done in 4.5 M DMSO for 3–5min at 22°C and in 4.5 M PROH for 3–5 min at 4°C.The results with regard to exposure to the freezing solutionsindicated that the loss of viability beyond an optimum is notdue solely to cryoprotectant toxicity, in particular not at4°C and not for DMSO. It is suggested that the temperatureand time of equilibration influence the degree of cryoprotectantpermeation and subsequent rehydration, which play a role indetermining freezing susceptibility in terms of ice formation.We conclude that both DMSO and, in contradiction to previousreports, PROH can be used perfectly adequately for ultrarapidfreezing on condition that concentration, and the temperatureand time of equilibration are controlled.     

NM23基因相关信号通路在肿瘤转移中作用机制的研究进展

浸润和转移是恶性肿瘤的重要特征,也给肿瘤的治疗带来困难,是预后不良的重要因素.转移抑制因子23(non-metastasis,NM23)基因是最早发现的抗肿瘤转移基因之一.现在已经发现NM23是一个基因家族,包括NM23-H1、NM23-H2等重要的基因家族成员.研究表明NM23基因表达与实体瘤转移抑制有关,在很多实体瘤中可以作为进展和预后的分子标记.随着对NM23基因调控肿瘤转移的分子机制的研究的进一步开展,已经发现了一些NM23肿瘤转移抑制通路上下游的相关调控分子,为进一步的信号通路研究创造了条件.本文概述了近年来对NM23基因转移抑制通路研究的新近展,提出了以后可能的研究方向和需要解决的关键问题.     

Paternal effects acting during the first cell cycle of human preimplantation development after ICSI.

BACKGROUND: The ability of human embryos to undergo normal development has been shown previously to be subject to strong paternal (sperm-derived) effects. This study was undertaken to determine whether paternal influences on human embryo quality are detectable as early as the first cell cycle after fertilization. METHODS: The quality of zygotes and cleaving embryos resulting from sibling donor oocytes fertilized by sperm from different patients were compared in a donor oocyte-sharing programme. RESULTS: Fertilizations with sperm from certain individuals repeatedly resulted in the formation of high proportions of zygotes with abnormal pronuclear morphology that subsequently tended to cleave slowly and to show extensive fragmentation and blastomere irregularities. This phenomenon was observed with oocytes from two different donors for each of these individuals and contrasted with normal developmental performance of embryos resulting from sibling oocytes fertilized by sperm from other men with similar basic sperm characteristics. Fertilization rates were not related to these differences. CONCLUSIONS: These data point to a very early onset of paternal effects that condition human embryo development. These effects may be both of genetic (related to the minor gene activity of the male pronucleus) or epigenetic (related to the sperm-derived oocyte-activating factor or sperm centrosome) origin.     

Laser-assisted zona pellucida thinning prior to routine ICSI

BACKGROUND: In MII oocytes showing difficult oolemma breakage, ICSI can cause an increase in the degeneration rate. This may be overcome by laser-assisted ICSI using a 5-10 micro m opening in the zona pellucida for injection. However, such a small opening might impair the hatching process, especially if assisted hatching is applied in addition. In order to prevent this, the present study used routine injection through an area of zona pellucida in which laser zona thinning had been applied, providing for both a reduced mechanical stress to the oocyte and assisted hatching. METHODS: This prospective study involved 100 cycles with 1016 MII oocytes. Conventional ICSI (control group) was compared with a modified laser-assisted ICSI (study group) in sibling oocytes. In the latter group oocytes were injected through an extended area of zona thinning. RESULTS: Degeneration rate was significantly lower in the study group (P < 0.004). There were no differences in fertilization, or formation and quality of blastocysts. In the study group embryo quality on day 2 was significantly better (P = 0.004) and herniation of day 5 blastocysts was increased (P = 0.005). Rates of implantation and pregnancy were not affected. However, on day 3 laser-assisted ICSI proved beneficial (P = 0.038) in terms of clinical pregnancy rate. CONCLUSIONS: The new method combines a less invasive ICSI technique with assisted hatching. Our preliminary data indicate that in addition to an improved oocyte survival, this new approach increases the hatching rate in vitro, which may explain the increase in pregnancy rate, at least in day 3 transfers.     

病毒性肝炎患者IL－1、IL－6和TNFα活性的检测

检测了甲、乙型病毒性肝炎患者外周血单个核细胞（ＰＢＭＣｓ）ＩＬ－１、ＩＬ－６和ＴＮＦα的诱生活性及其血清中活性。结果表明，乙型慢性活动性肝炎（ＣＡＨ）、乙型肝炎后肝硬化（ＨＣ）和乙型重型肝炎（ＳＨ）ＰＢＭＣｓ经脂多糖诱导后，ＩＬ－１活性分别为３５３１．１±８８２．７Ｕ／ｍ１、２７６９．７±７３０．４±Ｕ／ｍｌ和５３２９．３±１０８９．３Ｕ／ｍｌ，高于正常对照组（Ｐ＜０．０５或（０．０１）；ＩＬ－６诱生活性分别为３８．９０±１４．７５Ｕ／ｍ１、２．４５±１８．８５Ｕ／ｍｌ和７１．９５±２８．０５Ｕ／ｍｌ（与正常对照组相比，ｐ＜０．０５或＜０．０１）；ＴＮＦα诱生活性在乙型慢性迁延性肝炎（ＣＰＨ）、ＣＡＨ、ＨＣ和ＳＨ中分别为３３．２３±７．２５Ｕ／ｍｌ、６．９９±１．８４Ｕ／ｍ１、４．２９±２．１７Ｕ／ｍｌ和８６．７０±２４．１８Ｕ／ｍｌ，与对照组相比Ｐ＜０．０５或Ｐ＜０．０１。各型患者血清中ＩＬ－１、ＩＬ－６和ＴＮＦα活性均有不同程度的增高。文中对ＳＨ患者ＩＬ－１、ＩＬ－６和ＴＮＦα之间的相互关系进行了探讨。     

Pregnancies following blastocyst stage transfer in PGD cycles at risk for beta-thalassaemic haemoglobinopathies.

BACKGROUND: Preimplantation genetic diagnosis (PGD) usually involves blastomere biopsy 3 days post-insemination (p.i.), followed by genetic analysis and transfer of unaffected embryos later on day 3 or 4. We evaluate a strategy involving embryo biopsy on day 3 p.i., genetic analysis on day 4 and, following culture in blastocyst sequential media, transfer of unaffected embryos on day 5 p.i. METHODS: PGD cycles were initiated in 15 couples at risk of transmitting beta-thalassaemia major. Oocyte retrieval and ICSI were performed according to standard protocols. Embryo culture used blastocyst sequential media. Embryos were biopsied on day 3 p.i. using acid Tyrode's for zona drilling, and the single blastomeres were genotyped by a protocol involving nested polymerase chain reaction and denaturing gradient gel electrophoresis analysis. RESULTS: Forty of 109 (37%) embryos biopsied on day 3 p.i. developed to blastocysts by day 5 p.i., with at least one blastocyst available for transfer in 12 cycles (80%). Genotype analysis characterized 51/109 (47%) embryos unaffected for beta-thalassaemia major, of which 28 were blastocysts. Transfer of 37 day 5 p.i. embryos (blastocysts and non blastocysts) initiated eight clinical pregnancies. Implantation rate per embryo transferred was 12/37 (32%). CONCLUSIONS: Embryo biopsy on day 3, followed by delayed transfer until day 5 p.i. offers a novel and effective strategy to overcome the time limit encountered when performing PGD, without compromising embryo implantation.     

未成熟树突状细胞诱导实验性自身免疫性重症肌无力耐受的试验研究

目的 探讨未成熟树突状细胞(iDC)对实验性自身免疫性重症肌无力(EAMG)的影响。方法 用小剂量粒-巨噬细胞集落刺激因子(GM-CSF)体外诱导骨髓前体细胞，获得的iDC用乙酰胆碱受体(ACAR)负载后皮下接种同基因大鼠，3周后通过体外试验观察脾细胞对AChR的增殖反应，同时用AChR和完全弗氏佐剂(CFA)免疫大鼠，观察免疫7周后MG相关指标的改变。结果 小剂量GMCSF诱导所获得的iDC与成熟DC(mDC)相比，其表面MHCⅡ和共刺激分子CD80、CD86的表达低，摄取dextran-FITC的能力强但刺激同种T细胞增殖的能力弱。接种iDC、mDC、AChR负载的mDC的大鼠和对照组一样，体外脾细胞对AChR的刺激均强烈增殖，用AChR和CFA免疫后，产生明显的MG症状，重复电刺激出现明显衰减，血清AChRAb滴度明显增高，神经肌肉接头呈现典型的MG样改变；而接种AChR负载的iDC的大鼠脾细胞对AChR的刺激呈现明显的增殖抑制但对卵清蛋白的刺激仍强烈增殖，MG相关指标均未见明显改变。结论 骨髓前体细胞经小剂量GM-CSF诱导产生了iDC，AChR负载的iDC可诱导EAMG耐受。     

Neuropeptides stimulate production of interleukin-1β, interleukin-6, and tumor necrosis factor-α in human dental pulp cells

Objective:Orthodontic tooth movement causes inflammatory reactions in the periodontal membrane and dental pulp. It has been reported that substance P (SP) and calcitonin gene-related peptide (CGRP), both sensory neuropeptides, are manifested in the dental pulp of rats during experimental tooth movement, suggesting that they might be involved in the dental pulp inflammation during orthodontic tooth movement. However, the relationships between neuropeptides and pro-inflammatory cytokines have not been fully elucidated.Materials and methods:Human dental pulp (HDP) fibroblasts were prepared from 6 healthy young volunteers (3 males, 3 females; 15–25 years old) during the course of orthodontic treatment. HDP cells were incubated for 24 h in fresh medium containing 2% FCS in the presence of various concentrations of CGRP (10^–12 to 10^–4 M) and SP (10^–12 to 10^–4 M), and the levels of interleukin (IL)-1, IL-6, and tumor necrosis factor (TNF)- present in the media were determined using commercially available high-sensitivity enzyme-linked immunosorbent assay kit.Results:We examined the effects of stimulation by these neuropeptides on the production of inflammatory cytokines in HDP fibroblasts, and found that the levels of IL-1, IL-6, and TNF- increased in a time- and concentration-dependent manner. However, the neuropeptides did not act synergistically to increase cytokine secretion in HDP cells or significantly modify LPS-induced cytokine production by HDP cells.Conclusions:Our results suggest that human pulp fibroblasts may be involved in the progress of inflammation in pulp tissue during orthodontic tooth movement, as they produced large amounts of IL-1, IL-6, and TNF- following stimulation with neuropeptides.Received 2 March 2003; returned for revision 14 July 2003; accepted by M.J. Parnham 17 December 2003     

人血浆高密度脂蛋白对大鼠内毒素血症的治疗及防护作用研究

目的:观察人血浆高密度脂蛋白(HDL)对大鼠内毒素血症的治疗和防护效果。方法:采用鲎试剂法和放射免疫分析法于3个时点动态测定对照组、治疗组和防护组大鼠血浆内毒素(ET)水平和肿瘤坏死因子(TNF)浓度并观察血压及存活时间。结果:①输注ET后对照组大鼠血压进行性下降(P<0.01);治疗组大鼠输注HDL后其血压虽也降低但下降程度明显弱于对照组(P<0.01);防护组大鼠在输注ET后血压无明显下降(P>0.05)。治疗组及防护组大鼠存活时间均明显长于对照组(P<0.01)。②3组大鼠血浆ET水平在各时点均无明显变化(P>0.05)。③治疗组及防护组大鼠血浆TNFα水平于第3时点均明显降低(P>0.05)。结论:人血浆HDL能减轻或抑制内毒素血症大鼠血压下降并明显延长其存活时间,对内毒素血症具有良好的治疗和防护作用,此作用可能与其抑制TNF释放有关。