高糖通过活化半胱氨酸天冬氨酸蛋白酶3途径诱导人腹膜间皮细胞凋亡

目的 探讨高糖损伤人腹膜间皮细胞(HPMC)的机制。方法 (1)分离培养HPMC,以细胞形态、免疫细胞化学染色作细胞鉴定;(2)第2代HPMC分别经含1.5%、2.5%、4.25%葡萄糖的M199培养基培养48h后(以正常M199培养基和含4.25%甘露醇的M199培养基为对照),检测HPMC caspase-3活性和HPMC凋亡情况;(3)分别用25μmol/L、50μmol/L、和100μmol/LZ-VAD.fmk和4.25%葡萄糖共同刺激体外培养的HPMC(以DMSO和4.25%葡萄糖为对照),流式细胞仪观察HPMC凋亡情况,同时检测50μmol/L Z-VAD.fmk组和葡萄糖对照组caspase-3活性。结果 (1)HPMC caspase-3活性和凋亡率与葡萄糖浓度呈正相关(r分别为0.9506和0.9868,P<0.01),且HPMC caspase-3活性与凋亡率呈正相关(r=0.9860,P<0.01)。与M199对照组相比,2.5%葡萄糖组和4.25%葡萄糖组HPMC caspase-3活性和凋亡率显著增加(P<0.05),而1.5%葡萄糖组及4.25%甘露醇组HPMC caspase-3活性和凋亡率无显著差异(P>0.05)。(2)4.25%葡萄糖组caspase-3活性显著高于4.25%甘露醇组(P<0.05),但4.25%甘露醇组caspase-3活性与对照组相比无显著差异(P>0.05)。(3)与对照组相比,Z-VAD,fmk组HPMC凋亡率(P<0.05)和caspase-3活性(P<0.05)显著下降,且HPMC凋亡率与Z-VAD.fmk呈负相关(r=-0.8836,P<0.01)。结论 (1)高糖能以     

Differences in the hypothermic response to ethanol in rats selectively bred for oral ethanol preference and nonpreference

The duration of retention of tolerance to ethanol was tested in the alcohol-preferring (P) and alcohol-nonpreferring (NP) rats lines, using ethanol-induced hypothermia as a measure of tolerance. Rats received two injections of ethanol (3.5 g/kg) body wt, IP) and the time between the injections was 1, 2, or 3 days. When one day separated the two injections, tolerance to the hypothermic effect of a second “test” injection was found in both lines. When 2 or 3 days separated the two injections, the P line showed a loss of tolerance and the NP line showed sensitization to ethanol. Sensitization in the NP line grew stronger when the interval between injections was increased from 2 to 3 days. The duration of retention of tolerance to ethanol-induced hypothermia in the P line was shorter than has previously been reported for motor impairment in this line. It appears that the duration of tolerance retention in the P line depends on the test used to measure tolerance. Sensitization to ethanol in the NP line may be associated with low oral ethanol intake. This research was supported, in part, by grants AA08312, AA03243, and AA07611 from the PHS     

Characterization of T cell epitopes in αs1-casein in cow''s milk allergic, atopic and non-atopic children

BACKGROUND: One to two percent of infants suffer from IgE-mediated allergic reactions against cow's milk proteins. Most children develop clinical tolerance, but approximately 15% are still allergic by the age of 10 years. Little is known about the T cell epitopes in individual cow's milk protein in relation to allergy and tolerance. OBJECTIVE: To identify T cell epitopes in alphas1-casein, the most abundant milk protein, and to investigate T cell responses toward these epitopes in allergic, atopic and non-atopic children. METHODS: Allergen-specific T cell lines (TCLs) were derived from peripheral blood mononuclear cells of 11 cow's milk allergic, nine atopic and nine non-atopic children. T cell responses were measured to alphas1-casein and to overlapping peptides (18-mers), spanning the alphas1-casein molecule. Proliferation was determined by incorporation of (3)H-thymidine, and cytokine production (IL-10, IL-13 and IFN-gamma) was measured by ELISA. RESULTS: Four main regions (amino acid (AA) residues 43-66, 73-96, 91-114 and 127-180) in the alphas1-casein molecule were immunogenic to T cells, among which the AA residues 133-156 spanned the immunodominant part. Only subtle differences were found in peptide recognition between the subject groups. Some of the peptides induced slightly Th1- or Th2-skewed cytokine responses. The increased levels of IL-10 in response to alphas1-casein observed in TCLs from atopic children appeared not to be linked to recognition of specific IL-10-inducing epitopes. CONCLUSIONS: The immunodominant sequence in alphas1-casein is spanned by AA residues 133-156. Tolerance towards alphas1-casein in atopic children may be mediated by an overall induction of IL-10 and not by recognition of certain T cell epitopes. The identified T cell epitopes in children with cow's milk allergy may be useful targets in developing peptide immunotherapy.     

口服抗原对巨噬细胞共刺激分子CD80/CD86表达的影响

目的探讨口服抗原对巨噬细胞(M)共刺激分子CD80/CD86表达的影响及其在诱导妊娠免疫耐受中的作用。方法自然流产小鼠(CBA/J×DBA/2)分为免疫组和未免疫组,免疫组分别口服滋养细胞膜抗原(TMA2)和卵清蛋白(OVA),以正常妊娠小鼠(CBA/J×BALB/c)作为对照组。采用双标记流式细胞分析技术,分别检测各组小鼠脾脏及肠系膜淋巴结(MLN)CD80 M和CD86 M的表达。结果MLN内,未免疫组CD80 M表达明显高于对照组(P<0.05),而CD86 M含量明显低于对照组(P<0.001);TMA2免疫组CD80 M含量明显低于未免疫组(P<0.05);OVA免疫组CD86 M含量明显高于未免疫组(P<0.001)。脾脏内,CD80 M表达在各组间比较无显著差异;CD86M表达,未免疫组明显低于对照组(P<0.05),TMA2免疫组显著低于未免疫组(P<0.05),OVA免疫组则显著高于未免疫组(P<0.001)。结论流产的发生与M表面共刺激信号CD80/CD86异常有关;口服适当抗原可改变CD80/CD86 M表达模式,诱导妊娠免疫耐受的形成。     

Fecal stream is essential for adaptive induction of glucose-coupled sodium transport in the remnant ileum after total proctocolectomy

Our previous studies demonstrated that sodium glucose cotransporter 1 (SGLT-1) was induced in the remnant ileum of total colectomized rats via the action of factors other than hyperaldosteronism. The aim of the present study was to clarify whether fecal stream is required for the enhancement of SGLT-1-mediated sodium transport. Twenty-seven pairs of ileal tissues were obtained from the proximal and distal side, respectively, of loop ileostomy after total proctocolectomy. Mucosae were mounted in an Ussing chamber to evaluate glucose-coupled sodium transport. Levels of SGLT-1 mRNA in proximal and distal mucosae were compared by Northern blotting. Villous height and crypt depth were measured to test for correlations between mucosal structure and SGLT-1-mediated sodium transport or mRNA expression levels. Both glucose-coupled sodium transport and expression of SGLT-1 mRNA were significantly lower in distal mucosae relative to proximal mucosae. In distal mucosae, villous height, but not crypt depth, was significantly lower than in proximal mucosae, demonstrating a positive correlation between villous height and SGLT-1 function and expression. Comparative studies of proximal and distal mucosae demonstrated that in addition to hormonal changes, fecal stream is required for full induction of the sodium transport system (which includes SGLT-1-mediated transport) in the remnant ileum following total proctocolectomy. Presented in part at the Forty-Sixth Annual Meeting of The Society for Surgery of the Alimentary Tract, Chicago, Illinois, May 14–19, 2005 (poster presentation). This work was supported by Grants-in-Aid for Scientific Research 10557118 and 14657295 from the Ministry of Education, Science and Culture of Japan to K. Fukushima, and by Kanae Foundation to K. Fukushima.     

高血糖对急性颅脑伤预后的影响

对ＩＣＵ收治的６１例急性闭合性颅脑伤患者入院２４ｈ血糖水平与急性期格拉斯哥昏迷评分、颅脑伤不同类型及预后进行分析。结果提示：格拉斯哥昏迷评分与血糖水平经回归分析呈显著负相关（ｒ＝－０．９６４，Ｐ＜０．０１）；不同类型颅脑伤患者以脑挫裂伤组血糖最高。高血糖组患者病死率明显高于非高血糖组，死亡组患者血糖明显高于恢复不良组及恢复良好组。作者认为，血糖测定有助于了解颅脑伤的严重程度，同时可以估计患者的预后，在颅脑伤的救治过程中加强血糖监测是必要的。     

Pathogenesis of progressive muscular dystrophy: studies on free radical metabolism in an animal model

Evidence to suggest the presence of abnormal metabolism of oxygen free radicals in progressive muscular dystrophy is presented using an animal model. In the superficial pectoral muscles of dystrophic chickens, enzyme activities regulating the metabolism of oxygen free radicals, i.e., catalase, superoxide dismutases and glutathione peroxidase, were significantly elevated within 1 week of hatching. Activities of related enzymes, i.e., glutathione reductase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase were also elevated. In contrast, the specific activity of phosphofructokinase, the rate-limiting enzyme of the glycolytic pathway, was normal during the first 4-week period. These results suggest that there is an increased turnover of oxygen free radicals in the dystrophic muscle. This concept appears important in a further investigation of the pathogenesis and treatment of progressive muscular dystrophies.     

Intranasal treatment with ovalbumin but not the major T cell epitope ovalbumin 323–339 generates interleukin-10 secreting T cells and results in the induction of allergen systemic tolerance

BACKGROUND: Nasal administration of major peptide T cell epitopes gives contradictory data on the induction of peripheral tolerance. OBJECTIVE: To compare the prophylactic effect of intranasal treatment (INT) on the development of an allergic response, using either ovalbumin (OVA) or its major T cell epitope OVA 323-339 (OVAp). METHODS: BALB/c mice were treated intranasally with OVA or OVAp and subsequently immunized s.c. with OVA. Anti-OVA-specific antibody, T cell proliferation and cytokine responses were analysed. In an adoptive transfer model using OVAp specific TCR transgenic (Tg) T cells from D011.10 mice, in vivo tracking and characterization of transferred T cells in the cervical, inguinal and bronchial lymph nodes (BLN) and in the spleen were determined by FACS analysis. RESULTS: Prophylactic INT with OVA induced T cell tolerance towards subsequent OVA s.c. immunizations, inhibiting OVA specific T cell proliferation, IgE and IgG1 production, in contrast to INT with OVAp, which was unable to induce tolerance. In vivo analysis of transferred OVA-specific TCR Tg T cells showed that INT with OVA induced a preferential activation of T cells in BLN, as opposed to a broad, systemic activation with OVAp. In vivo, OVAp INT led to faster and more sustained cell division cycles than OVA INT. Ex vivo, tolerance to OVA was associated with the generation of IL-10 secreting CD4(+) T cells in BLN of OVA-treated mice only. CONCLUSION: INT with OVA but not with OVAp led to regional (as opposed to systemic) T cell activation and the induction of IL-10 secreting CD4(+) T cells in BLN, potentially critical steps in the induction of T cell-specific tolerance via the nasal route.     

TIA对后继脑梗死神经保护作用的临床研究

目的探讨预先短暂性脑缺血发作(TIA)是否对后继脑梗死具有神经保护作用。方法根据脑梗死患者是否预先发生同侧的TIA分为TIA组和无TIA组,分别从TIA持续时间、发作次数、与脑梗死间隔时间及梗死部位几方面同无TIA组比较,采用ADL评分标准,评定患者预后恢复情况。结果Ⅰ级预后预先有TIA的占72%,无TIA的占63%,二者相比差异无显著性(X~2=3．31,P＞0．05)。但脑梗死前有TIA且持续时间在5～20min、发作次数2～3次,与脑梗死间隔不超过1周者,其Ⅰ级预后较多,同无TIA组相比较差异有显著性(P＜0．05)。结论脑梗死前发生的TIA有可能对人脑产生缺血耐受作用。TIA的神经保护性与其发作持续时间、发作次数及与脑梗死间隔时间有一定关系。     

Relationship between maternal glycaemia and birth weight in glucose-tolerant women from different ethnic groups in New Zealand.

AIM: The aim of this study was to compare the population attributable fraction(PAF) for a large baby (> or =4 kg) due to glycaemia, weight and smoking in glucose-tolerant women from different ethnic groups. METHODS: A retrospective review of screening for gestational diabetes (GDM)and associated birth weight was undertaken in New Zealand European (n= 529), Maori (n= 540) and Pacific (n= 916) women. The proportion with a large baby was compared by 1-h post 50-g glucose challenge test tertile and maternal weight tertile. RESULTS: Large babies were more common from Pacific and European than Maori women (24.3%, 18.8%, 8.9%, respectively; P<0.001). Birth weight increased significantly with increasing glucose among Pacific women (P<0.001) even after adjusting for maternal weight and other confounders. The risk of having a large baby was 2.56 (1.82-3.60)-fold greater in women in the highest maternal weight tertile (> or =84 kg), with a significantly greater PAF in Pacific women(27.2%, 12.9%, 16.4%, respectively; P<0.001). The odds ratio (OR) of having a large baby increased with even mildly elevated maternal 1-h glucose concentrations [OR for 5.6-6.2 mmol/l: 1.54 (1.11-2.14); for > or =6.3 mmol/l: 2.06 (1.50-2.82)], with no ethnic differences in PAF (11.1-11.8%, 16.7-18.7%, respectively). Smoking and being Maori were associated with smaller babies. CONCLUSIONS: Increased maternal weight and glycaemia are associated with a greater proportion of large babies among glucose-tolerant women. Growth of Pacific babies may be more sensitive to a higher maternal glucose when the mother is obese.