转hCTLA4Ig树突状细胞诱导T细胞免疫耐受的实验研究

目的　通过逆转录病毒载体将人CTLA4Ig转染DCs ,探讨转人CTLA4Ig(hCTLA4Ig)树突状细胞 (DCsRev)诱导T细胞免疫耐受的可能性。方法　通过重组逆转录病毒将目的基因hCTLA4Ig转染到大鼠骨髓来源的DCs中 ,通过流式细胞检测目的基因hCTLA4Ig表达及DCs表面分子的改变 ;通过混合淋巴细胞反应 (MLR)检测DCsRev抑制T细胞免疫反应的能力。 结果　重组逆转录病毒转染DCs的最大效率为 91 2 5 % ;在功能上 ,DCsRev不但丧失了刺激MLR的能力 ,并且能够强烈抑制MLR中反应T细胞的增殖 ,而且抑制率与加入DCsRev的数量和DCsRev预处理反应T细胞的时间长短有关。具体来说 ,DCsRev数量在 10 3 ～ 10 4之间时 ,抑制率与剂量呈正相关 ,最高为 71 96%。而当DCsRev数量达到 5× 10 4抑制率下降为 5 9 2 %。在 12～ 48h之间 ,随着预处理时间的延长 ,抑制率却不断下降 ,预处理 12h抑制率最高 ,为 99 6%。但不做预处理 ,在反应开始时同时加入DCsRev ,则抑制率明显降低 ,仅为 5 9 2 %。对腹腔注射DCsRev大鼠脾T淋巴细胞体外分析表明 ,DCsRev也能在动物体内诱导耐受 ,但这种免疫耐受状态不能维持终身。结论　通过逆转录病毒载体将人CTLA4Ig转染DCs,不但DCs表面CD86分子被CTLA4Ig有效的封闭 ,并且能够诱导抗原特异性T细胞的免疫耐受     

Synthesis of deramciclane* labeled with tritium in various positions

[(1R)‐endo]‐(+)‐3‐bromocamphor was dehalogenated with tritium gas to [3‐³H]camphor and via [3‐³H]phenylborneol converted to [3‐³H]deramciclane isolated as the fumarate salt (specific activity 51.8 GBq/mmol). This three step synthesis from [3‐³H]camphor gave an overall yield of 22%. Benzyloxy‐acetic acid methyl ester was reduced with sodium‐borotritide to 2‐benzyloxy‐ethanol‐[1‐³H], and through a four step procedure was converted to 2‐dimethylaminoethyl‐[2‐³H] chloride. The latter was condensed with the sodium derivative of 2‐phenylborneol giving rise to [2‐dimethylamino‐[2‐³H]ethoxy]deramciclane isolated as the fumarate (specific activity 8.177 GBq/mmol). This six step synthesis from [³H]NaBH₄ gave an overall yield of 6%. Copyright © 2005 John Wiley & Sons, Ltd.     

红细胞调控白细胞免疫功能新的自然实验研究体系

目的用血液免疫反应路线图实验体系评估红细胞在白细胞免疫活性中的作用。方法将0·3ml血浆加入0·2ml全血细胞悬液(全血细胞组)或0·2ml白细胞悬液(白细胞组)中,37℃温育1h,用免疫酶联法测定IL-8和IL-12水平,流式细胞仪测定白细胞膜CD4、CD8、CD35和CXCR4表达量。结果全血细胞组IL-8和IL-12水平(分别为5·96±4·26、9·84±2·23ρB·pg-1·ml-1)明显低于白细胞组(分别为15·09±9·86、13·59±3·69ρB·pg-1·ml-1,P<0·05),淋巴细胞CD4、CD35、CXCR4表达量(分别为37·79±12·00、154·66±70·00、34·40±20·45)明显高于白细胞组(分别为18·54±11·32、83·26±35·99、16·69±11·09,P<0.01),粒细胞CD35表达量(603·63±257·64)明显高于白细胞组(384·86±174·16,P<0.01)。成人全血细胞组淋巴细胞和粒细胞CD35和CXCR4表达量明显高于脐血全血细胞组(P<0·05或P<0·01)。结论红细胞是白细胞(包括T淋巴细胞、B淋巴细胞、NK细胞、树突状细胞、粒细胞等)免疫功能的调控者和指导者,脐血红细胞免疫调节功能明显下降;本研究为红细胞免疫调控活性测定提供了新的近似自然的方法。     

Are hospitals prepared to support newborn survival? – an evaluation of eight first‐referral level hospitals in Kenya*

Objective To assess the availability of resources that support the provision of basic neonatal care in eight first‐referral level (district) hospitals in Kenya. Methods We selected two hospitals each from four of Kenya’s eight provinces with the aim of representing the diversity of this part of the health system in Kenya. We created a checklist of 53 indicator items necessary for providing essential basic care to newborns and assessed their availability at each of the eight hospitals by direct observation, and then compared our observations with the opinions of health workers providing care to newborns on recent availability for some items, using a self‐administered structured questionnaire. Results The hospitals surveyed were often unable to maintain a safe hygienic environment for patients and health care workers; staffing was insufficient and sometimes poorly organised to support the provision of care; some key equipment, laboratory tests, drugs and consumables were not available while patient management guidelines were missing in all sites. Conclusion Hospitals appear relatively poorly prepared to fill their proposed role in ensuring newborn survival. More effective interventions are needed to improve them to meet the special needs of this at‐risk group.     

Synthesis,AM<Subscript>1</Subscript> calculation,and biological studies of thiopyran-4-one and their azine derivatives

Some novel N(1)-arylidene-N(2)-cis-2,6-diphenyltetrahydrothiopyran-4-one azine derivatives were synthesised and their antibacterial activity against Streptococcus faecalis, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus and antifungal activity against Candida-6, Candida-51, Aspergillus niger, and Aspergillus flavus were evaluated.     

散发性结直肠癌中SMAD4基因的突变

目的观察SMAD4基因在散发性结直肠癌中的突变，并探讨其对散发性结直肠癌发生发展的意义。方法对本院83例散发性结直肠癌患者，应用聚合酶链反应-单链构象多态性（PCR—SSCP）分析癌组织中SMAD4基因各外显子的突变情况，基因突变率与临床病理参数间采用，检验；应用多态性微卫星标记研究SMAD4基因所在18q21区的杂合性缺失。结果SMAD4基因在83例散发性结肠癌的患者中共有9例发生突变，平均突变率为10．8％。经Χ^2检验SMAD4基因突变率在肿瘤的Dukes分期及有无远处转移间差异有统计学意义（P〈0．05），而与肿瘤的分化程度、发生部位及患者的性别差异无统计学意义（P〉0．05）。18q21区的杂合性缺失率为62．71％，其中发生突变的9例均存在18q21区的杂合性缺失。结论SMAD4基因的突变可能介导了散发性结直肠癌后期的发生发展。     

Histopathological characterisation of effects of the mouse Pax6 missense mutation on eye development

Mutations in PAX6/Pax6 lead to a variety of ocular anomalies in humans and mice. The aim of the study was to characterise the ocular abnormalities caused by the missense Pax6^Leca4 mutation and compare them to published observations on Pax6 alleles that are functionally equivalent to Pax6⁻ null alleles (such as Pax6^Sey and Pax6^Sey-Neu) and human inherited eye diseases. Ocular features of homozygous Pax6^Leca4/Leca4 and heterozygous Pax6^Leca4/+ embryos at E12.5-E18.5, heterozygous Pax6^Leca4/+ young mice at P18 and heterozygous Pax6^Leca4/+ adults at 12 weeks were analysed histologically with their wild-type Pax6^+/+ littermates. Homozygous Pax6^Leca4/Leca4 fetuses died perinatally with no eyes although an optic cup rudiment with pigmented cells developed. Pax6^Leca4/+ mice were microphthalmic and a range of other severe ocular phenotypes affected both the anterior and the posterior segments. In contrast to Pax6^+/−, the Pax6^Leca4/+ eyes had no goblet cells in the corneal epithelium, the iris was not hypoplastic and there was no lens-corneal epithelial plug. However, microphthalmia was more severe, corneal vascularisation occurred earlier (during fetal stages), pigmented cells were present in the vitreous and corneal stroma and the ciliary body was malformed or abnormal. These results show that, although Pax6^Leca4/+ lacked some eye abnormalities commonly seen in Pax6^Sey/+ and Pax6^Sey-Neu/+ eyes, in most respects their eyes were more severely affected. These differences probably reflect both differences between the Pax6^Leca4 and the Pax6^Sey-Neu mutations and differences in modifier gene expression in different genetic backgrounds. The presence of pigmented cells in the cornea is a novel observation. Some Pax6^Leca4/+ ocular abnormalities were similar to those present in human Peters' anomaly and persistent hyperplastic primary vitreous (PHPV) so Pax6^Leca4/+ mice provide a useful model for some inherited eye diseases.     

Flavocoxid is as effective as naproxen for managing the signs and symptoms of osteoarthritis of the knee in humans: a short-term randomized,double-blind pilot study

Flavocoxid (Limbrel), a proprietary mixture of flavonoid molecules (baicalin and catechin), was tested against a traditional nonsteroidal anti-inflammatory drug, naproxen, for the management of the signs and symptoms of moderate osteoarthritis (OA) in humans. Discomfort and global disease activity were used as the primary end points, and safety assessments were also taken for both treatments as a secondary endpoint. In this double-blind study, 103 subjects were randomly assigned to receive either flavocoxid [500 mg twice daily (BID)] or naproxen (500 mg BID) in a 1-month onset of action trial. Outcome measures included the short Western Ontario and McMaster University Osteoarthritis Index, subject Visual Analogue Scale for discomfort and global response, and investigator Visual Analogue Scale for global response and fecal occult blood. Both flavocoxid and naproxen showed significant reduction in the signs and symptoms of knee OA (P ≤ .001). There were no statistically detectable differences between the flavocoxid and naproxen groups with respect to any of the outcome variables. Similarly, there were no statistically detectable differences between the groups with respect to any adverse event, although there was a trend toward a higher incidence of edema and nonspecific musculoskeletal discomfort in the naproxen group. In this short-term pilot study, flavocoxid was as effective as naproxen in controlling the signs and symptoms of OA of the knee and would present a safe and effective option for those individuals on traditional nonsteroidal anti-inflammatory drugs or cyclooxygenase-2 inhibitors. A low incidence of adverse events was reported for both groups.     

IL-4与IL-10联合诱导异种骨移植免疫耐受的体外研究

目的　探讨 IL - 4和 IL - 10在诱导异种骨移植免疫耐受中的作用。方法　反应细胞为 BAL B/c小鼠脾淋巴细胞 ,刺激细胞为新西兰白兔血淋巴细胞 ,刺激抗原为兔骨上清液。采用经典的混合淋巴细胞培养法及骨上清液与淋巴细胞混合培养法作为异种骨移植的体外实验模型。在各培养液中分别加入 IL - 4、IL - 10及两者联合应用 ,通过测定其 3H- Td R掺入率 ,观察不同细胞因子对刺激淋巴细胞增殖的影响。结果　无论在细胞刺激组还是骨上清液刺激组 ,IL- 4对淋巴细胞增殖均有显著抑制作用 (P<0 .0 0 1和 P<0 .0 5 ) ,IL- 10未表现出抑制作用 (P>0 .0 5 )。在两组 IL- 4和 IL - 10联合应用均产生比 IL - 4单独应用更为明显的细胞增殖抑制作用 (P<0 .0 0 1和 P<0 .0 5 )。结论　 IL - 4对由细胞或骨上清液刺激产生的淋巴细胞增殖均有很好的抑制作用 ,IL- 10没有表现出抑制作用 ;IL- 4与 IL- 10联合应用有协同抑制作用。