rhGM—CSF／IL—3融合蛋白对人骨髓造血祖细胞体外培养的研究

研究了ｒｈＧＭ－ＣＳＦ／ＩＬ－３融合蛋白对人骨髓造血祖细胞（ＣＦＵ－ＧＭ、ＣＦＵ－ＧＥＭＭ、ＢＦＵ－Ｅ）集落形成的影响，结果表明ｒｈＧＭ－ＣＳＦ／ＩＬ－３能显著促进ＣＦＵ－ＧＭ、ＣＦＵ－ＧＥＭＭ、ＢＦＵ－Ｅ集落的形成，分别与ＧＭ－ＣＳＦ、ＩＬ－３单独或联合用药比较，差异有显著性（Ｐ〈０．００１），ＣＦＵ－ＧＭ、ＣＦＵ＝ＧＥＭＭ、ＢＦＵ－Ｅ集落的形成对融合蛋白均有剂量依赖性，培养体系浓度在５￣１０ｎ     

rhIL—6促进致死性辐射损伤小鼠凝血及早期造血功能的恢复

目的：观察ｒｈＩＬ－６对小鼠造血功能的影响。方法：应用致死性６０Ｃｏ－γ射线辐射Ｂａｌｂ／ｃ小鼠，造成骨髓型辐射损伤模型。照射后用ｒｈＩＬ－６皮内注射给药６ｄ（２μｇ／只，２次／ｄ），通过检测出凝血时间（ＢＴ，ＣＴ）、凝血酶原时间（ＰＴ）、外周血ＰＬＴ和ＷＢＣ，以及骨髓造血细胞体外培养ＣＦＵ－Ｍｉｘ和脾脏ＣＦＵ－Ｓ的数目，研究了ｒｈＩＬ－６对骨髓型辐射损伤小鼠早期造血功能恢复的效应，并进行了机理的探讨。结果：ｒｈＩＬ－６处理组小鼠的ＢＴ，ＣＴ和ＰＴ均较对照组显著缩短（Ｐ＜０．０１）；外周血ＰＬＴ和ＷＢＣ分别较对照组提高１３０％和１６５％。骨髓造血细胞体外培养ＣＦＵ－Ｍｉｘ和脾脏ＣＦＵ－Ｓ计数，均显著高于对照组（Ｐ＜０．０１）。结论：ｒｈＩＬ－６具有促进受辐射小鼠早期造血细胞恢复的生物学效应，可减轻辐射所致骨髓损伤和促进造血干／祖细胞（ＨＳＣ／ＨＰＣ）的增殖、分化，有利于骨髓造血功能的恢复。     

重组人IL－6对大鼠急性白血病细胞体外增殖的抑制

我们已报道重组大鼠ＩＬ－３和小鼠ＧＭ－ＣｓＦ对ＬＴ１２白血病细胞的增殖有不同程度的抑制活性。本研究发现重组人ＩＬ－６在１０００～５０００Ｕ／ｍｌ呈剂量依赖性抑制ＬＴＩ２白血病祖细胞集落形成及ＤＮＡ合成。５０００Ｕ／ｍｌ对ＣＦＵ－Ｌ的抑制率达５２．９％，而对ＤＮＡ合成则为４１．３％，此外，ＩＬ－６还明显增加ＩＬ－３和ＧＭ－ＣＳＦ对ＬＴ１２细胞的抑制活性，对ＣＦＵ－Ｌ的抑制强弱依次为ＩＬ－６＋ＧＭ－ＣＳＦ、ＩＬ－３＋ＧＭ－ＣＳＦ、及ＩＬ－６＋ＩＬ－３，ＩＬ－６＋ＩＬ－３＋ＧＭ－ＣＳＦ三因子的结合并不能增加抑制效应；对ＤＮＡ合成的抑制作用为ＩＬ－６＋ＩＬ－３＋ＧＭ－ＣＳＦ＞ＩＬ－６＋ＧＭ－ＣＳＦ＞ＩＬ－３＋ＧＭ－ＣＳＦ＞ＩＬ－６＋ＩＬ－３。提示上述造血因子除刺激正常造血外，在白血病的治疗中可能具有一定意义。     

短程大剂量格拉诺赛特动员自体外周血造血干细胞？…

目的 报导短程大剂量格拉诺赛（ｒｈＧ－ＣＳＦ）动员外周血造血干细胞，进行自体外周血造血干细胞移植（ＡＰＢＳＣＴ）治疗急性白血病。方法 ｒｈＧ－ＣＳＦ，５００μｇ／ｄ皮下注射４天，第５、６天单采外周血单个核细胞（ＭＮＣ），行ＣＦＵ－ＧＭ培养，ＣＤ３４＾＋细胞计数。予ＭＡＣ方案预处理，回输自体外周血造血干细胞（ＡＰＢＳＣ）。结果 动员后ＣＦＵ－ＧＭ明显增高，ＣＤ３４＾＋细胞增多，骨髓造血功能重建迅速。     

固本生血丸对人骨髓造血祖细胞体外增殖的影响

应用造血祖细胞体外培养法研究了中药复方制剂——固本生血丸对人骨髓造血祖细胞体外集落形成的影响。结果表明：该制剂在０．０１～１０μｇ／ｍｌ浓度范围内与细胞因子（ＧＭ－ＣＳＦ或ＥＰＯ）协同促进正常骨髓和再障骨髓ＣＦＵ－ＧＭ、ＨＣＦＵ－ＧＭ，ＣＦＵ－Ｅ和ＢＦＵ－Ｅ的集落形成；作用高峰在１０μｇ／ｍｌ浓度上；当超过该浓度时，集落数量增加不明显（Ｐ＞０．０５），而单独应用该制剂时无集落形成。     

脐血红细胞生成素和白细胞介素6水平检测

目的 :红细胞生成素 (ＥＰＯ)特异性作用于红系造血细胞 ,刺激红系祖细胞增殖、分化、成熟 ,是红细胞系统造血必不可少的造血刺激因子。白细胞介素 6(ＩＬ - 6)促进造血干细胞从Ｇ0 期进入Ｇ1 期 ,与ＥＰＯ、ＧＭ -ＣＳＦ及ＩＬ - 3协同作用可促进红系爆增生式集落形成单位 (ＢＦＵ -Ｅ)、粒细胞巨噬细胞集落形成单位 (ＣＦＵ -ＧＭ)、巨核细胞集落形成单位 (ＣＦＵ -ＭＫ)和多向性祖细胞集落形成单位 (ＣＦＵ -ＧＥＭＭ)形成。体外实验证实脐血浆具有刺激造血的活性。本实验检测正常足月新生儿脐血浆的ＥＰＯ和ＩＬ - 6水平以了解ＥＰＯ与…     

固本生血丸对内骨髓造血祖细胞体外增殖的影响

应用造血祖细胞体外培养法研究了中药复方制剂－－固本生血丸对人骨髓造血祖细胞估外集落形成的影响。结果表明：该制剂在０．０１－１０μｇ／ｍｌ浓度范围内与细胞因子（ＧＭ－ＣＳＦ）或ＥＰＯ）协同促进正常骨髓和再障骨髓ＣＦＵ－ＧＭ、ＨＣＶＦＵ－ＧＭ，ＣＦＵ－Ｅ和ＢＦＵ－Ｅ的落形成；作用高峰在１０μｇ／ｍｌ浓度上；当超过该浓度时，集落数量增加不明显，而单独应用该制剂时无集落形成。     

人参总皂甙诱导造血生长因子生成的实验研究

为探讨人参“补气生血”的现代生物学机理，采用造血祖细胞体外培养，造血生长因子生物活性检测等技术，研究人参总皂甙（ＴＳＰＧ）对造血生长因子的诱导生成作用。结果表明，经ＴＳＰＧ诱导分别制备的脾细胞、巨噬细胞、成纤维细胞和骨髓基质细胞的培养上清对髓系造血祖细胞（ＢＦＵ－Ｅ、ＣＦＵ－ＧＭ、ＣＦＵ－ＭＫ）的体外集落增殖均有显著刺激作用。ＴＳＰＧ也能促进脾细胞、成纤维细胞和巨噬细胞分泌ＩＬ－６；刺激脾细胞产生     

脐血清对培养人造血祖细胞集落生长的影响

目的：研究脐血清对培养人造血祖细胞集落生长的影响。方法：采用半固体细胞培养法。结果：在正常足月胎儿脐血清中有较高粒细胞集落刺激因子（Ｇ－ＣＳＦ）和粒－巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）活性，单用脐血清作刺激因子来源，在体外进行粒系造血祖细胞培养，可产生１４８１±８８７个粒系集落（ＣＦＵ－Ｇ），而植物血凝素刺激的单个核细胞条件培养液为５９０±２８８个ＣＦＵ－Ｇ，差别非常显著（Ｐ＜０００１）。脐血清组与３０ｎｇｒｈＧ－ＣＳＦ组所产生１５２０±３４５个ＣＦＵ－Ｇ相比，差别无显著（Ｐ＞００５）。并且我们还观察到脐血清组培养随时间延长有较多粒－巨噬细胞集落（ＣＦＵ－ＧＭ）生长，有些可出现混合爆式集落。结论：脐血清富含多种造血生长因子，并且Ｇ－ＣＳＦ水平较高     

脐带血清对骨髓粒—巨噬细胞集落形成的影响

在骨髓细胞半固体琼脂培养体系中，研究脐带血清（ＣＢｓ）对骨髓粒—巨噬细胞集落形成单位（ＣＦＵ－ＧＭ）的影响，单纯应用ＣＢｓ培养可使骨髓ＣＦＵ－ＧＭ（ｘ±ｓ为２６．０３７５±１０．２３２７）显著的高于正常成人外周血清（ＰＢｓ）组（ｘ±ｓ为１１．５９１７±３．６０４７；Ｐ＜０．００１）。ＣＢｓ加粒－巨噬细胞刺激因子，ＧＭ－ＣＳＦ组ＣＦＵ－ＧＭ（ｘ±ｓ为１１９．０７９２±３５．０９９１）是单纯ＣＢｓ组的４．５７倍；而ＰＢｓ加ＧＭＣ－ＳＦ组（ｘ±ｓ为９７．１２５±２６．７５５９）是单纯ＰＢｓ组的８．３８倍。有白细胞介素－６（ＩＬ－６）存在的ＣＢｓ组，ＣＦＵ－ＧＭ（ｘ±ｓ为３１．９３３３±１０．９１４４）明显的高于单纯ＣＢｓ组（Ｐ＜０．０２）。提示ＣＢｓ中含有较高水平的内源性ＧＭ－ＣＳＦ。     

蓖麻毒素诱导U937细胞分泌IL-6和IL-8的作用

目的：进一步研究蓖麻毒素是否具有诱生Ｕ９３７细胞分泌细胞因子作用。方法：采用ＭＴＴ法检测了蓖麻毒素对Ｕ９３７细胞的毒性，并采用ＥＬＩＳＡ法测定细胞２上清中的Ｉ－６和ＩＬ－８。结果：蓖麻毒素对Ｕ９３７细胞的生具有时间效应及剂量效应。随着作用时间的延长。诱生２种细胞因子的量逐渐增加；随蓖麻毒素剂量的增加，诱生２种细胞因子的量均减少。结论：蓖麻毒素诱导２种细胞因子的产生为其抗癌应用或其它作用提供理论依据     

一组抗人重组IL－6单克隆抗体的制备及鉴定

以基因重组的人ＩＬ－６免疫了Ｂａｌｂ／ｃ小鼠，利用小鼠杂交瘤技术，建立了２０余株分泌抗人重组ＩＬ－６单克隆抗体的杂交瘤，并对其中的Ｃ＿（９２）、Ｃ＿（９７）、Ｃ＿（２０３）、Ｃ＿（２２０）进行了较系统的鉴定，其抗体类别均为ＩｇＧ，亚类分别为ＩｇＧ＿（２ａ）、ＩｇＧ＿１、ＩｇＧ＿（２ａ）和ＩｇＧ＿（２ａ）。它们均特异性地识别ＩＬ－６，而与多种其它因子和无关蛋白无交叉反应。免疫转印结果显示，各单抗均能识别分子量为１７ｋＤ的ＩＬ－６单一条带，并具有中和ＩＬ－６生物活性的效应。     

延髓中缝核5-羟色胺对清醒大鼠胃运动的影响

本工作采用应力传感器记录胃窦的运动,观察向清醒大鼠延髓中缝核注射1微升生理盐水中含6.25μg、12.5μg和25μg三种不同剂量的5-羟色胺(5HT)以及腹腔注射对氯苯丙氨酸(pCPA)或延髓中缝核注射5,6-双羟色胺(5,6DHT)以减少内源性5HT对胃运动的影响。实验结果表明:6.25μg 5HT有兴奋胃窦运动的趋势,12.5μg和25μg 5HT使胃窦收缩活动加强。将上述三种剂量的5HT注入外周静脉均不影响胃的运动。腹腔注射pCPA后第四天和延髓中缝核注射5,6DHT后第五天,抑制大鼠胃运动。此时,颈髓、延髓、中桥脑、间脑5HT含量均明显下降。提示延髓中缝核5HT参与胃运动的中枢性调节。     

Orientation of major histocompatibility (MHC) genes relative to the centromere of human chromosome 6

Linkage analysis of the data obtained from a three-generation Dutch family segregating for genetic variants of centromeric heterochromatic region in the band pi 1 on chromosome 6 (6ph), major histocompatibility (MHC) genes HLA-A, HLA-B and HLA-C, glyoxalase I (GLO) and phosphoglucomutase-3 (PGMg) showed that the genetic distance between the HLA gene cluster and 6ph is about 6 cM (at an estimated peak lod score of 3.466), that GLO is closer than HLA to the centromere and that PGM₃ is probably not situated on the same chromosomal arm as HLA.     

男性老年骨质疏松症患者血清IL-6、BGP和T含量变化及临床意义

目的:探讨老年骨质疏松症患者血清白细胞介素-6(IL-6)、骨钙素(BGP)和睾酮(T)含量的变化及临床意义。方法:应用放射免疫分析对46例老年骨质疏松症患者进行了血清IL-6、BGP和T水平测定,并与35名正常健康人作比较。结果:老年骨质疏松症患者血清IL-6水平明显高于正常健康人(P<0.01),而BGP、T水平明显低于正常人(P<0.01)。结论:测定老年骨质疏松症患者血清中IL-6、BGP和T水平为临床诊断治疗提供了一个有价值的数据。     

支气管哮喘患儿治疗前后血清GM-CSF、IL-8和IL-6联检的临床意义

目的：探讨了支气管哮喘患儿血清GM—CSF、IL-8和IL-6水平的变化及意义。方法：应用放射免疫分析对32例支气管哮喘患儿进行了血清GM—CSF、IL-8和IL-6水平测定，并与30例正常健康儿作比较。结果：支气管哮喘患儿血清GM—CSF、IL-8和IL-6水平非常显著地高于正常儿组（P〈0．01）。经治疗3个月后则与正常儿比较无显著性差异（P〉0．05）。结论：血清GM—CSF、IL-8和IL-6水平的异常升高足支气管哮喘患儿发病的病理因素之一．有重要的临床价值。     

Lung epithelial H292 cells induce differentiation of immature human HMC-1 mast cells by interleukin-6 and stem cell factor

BACKGROUND: Immature mast cells migrate into tissues where they differentiate into mature mast cells under the influence of local factors. In the airways of asthmatics increased numbers of chronically activated mast cells are located nearby the airway epithelium. OBJECTIVE: The aim of this study was to evaluate whether and, if so, which products released by epithelial cells may affect mast cell proliferation and differentiation. METHODS: We performed in vitro studies using the human lung mucoepidermoid carcinoma-derived H292 cell line and the immature human mast cell line, HMC-1. Proliferation was assessed by 3H-thymidine incorporation. Differentiation of HMC-1 cells was inferred from tryptase production. RESULTS: Exposure of HMC-1 cells to medium conditioned for 48 h by H292 cells resulted in a reduction of proliferation with 65 +/- 4.9% (mean +/- SEM, n = 9) at day 5. Culturing HMC-1 cells for 8 days in the presence of H292-conditioned medium resulted in morphological changes indicative of differentiation, and in a 3.0 +/- 0.4-fold increase of tryptase production (P = 0.0039, n = 9). Conditioned medium from H292 cells that were stimulated by LPS also inhibited HMC-1 proliferation. Inhibitory antibodies against two mediators from H292 cells, interleukin-6 (IL-6) and stem cell factor (SCF), abolished the increase in HMC-1 tryptase production induced by H292-conditioned medium. Recombinant human (rh) IL-6, but not rhSCF, reduced HMC-1 proliferation with 44% and 13% at day 3 and 5, respectively. Surprisingly, rhIL-6 did not increase HMC-1 tryptase production significantly whereas incubation with rhSCF did (1.5 +/- 0.1-fold, P = 0.002, n = 10) although the increase was less than observed for conditioned medium. CONCLUSION: Epithelial-derived IL-6 and SCF are implicated in differentiation of HMC-1 cells but additional factors are not excluded. As activated primary bronchial epithelial cells also express IL-6 and SCF, it should be considered that these cells are involved in mast cell differentiation within the airways, particularly in diseases where epithelial cells are activated, such as asthma.     

On the causation of diabetes mellitus (effect of saliva on blood glucose levels in oral glucose tolerance tests)

17 male patients, aged 25 – 58, completed four sets of blood glucose estimations, fasting at 30 minutes, 60 minutes and 120 minutes after a 50 g. oral glucose load. The study was so designed that saliva was added to the glucose meal in one set of observations and excluded from the other. Results based on 136 observations on blood glucose showed that at the time intervals, namely, 30 minutes and 60 minutes, the blood glucose levels were significantly lower when the glucose meal was admixed with saliva than when it was excluded, although there was no significant difference in the mean fasting levels. The difference at 120 minutes did not reach 1% level of significance. The differences in blood glucose levels may be due to the saliva retarding gastric emptying. In the case of meals with no saliva, a rapid “dumping” of glucose takes place as a result of increased gastric emptying rate and this may give rise to sucrose-induced hyperinsulinism, which, if prolonged over extended periods, may result in insulin-resistance. The slower rise of blood glucose after meals admixed with saliva will not have this effect because insulin-release and glycaemia would go hand in hand. This may possibly explain the increased incidence-rates of diabetes mellitus in individuals used to “meal-scamping” as noted in the companion study (1), as compared with “meal-chewers”.It has been proposed that a sudden and rapid rise of insulin levels in the blood, whether due to a sudden hyperglycaemia due to rapid gastric emptying and resultant “dumping” of sucrose, or due to secretin-release, may give rise to insulin anti-body production (1), setting up the vicious cycle — hurried meals — rapid gastric emptying accompanied by hyperglycaemia and hyperchlorhydria (2,3) — secretin-release (4) — hyperinsulinism (5,6) — hypoglycaemia, persistence of insulin — insulin resistance and insulin antibodies (7,8) — islet over-activity and islet-cell exhaustion (7–9).Yudkin et al (10) observed that out of their 13 subjects, only 6 showed hyperinsulinism after a sucrose meal. They did not find out the reasons for this susceptibility of only some persons to develop hyperglycaemia and hyperinsulinism after a high sucrose diet. It has been propsed by me (1,11) that this susceptibility may be due to the pattern of eating: persons who masticate their food well, so that a mucus-rich saliva mixes with the meal, do not develop this and those who do not masticate their food well, so that saliva does not mix with the meal, develop hyperglycaemia and resultant hyperinsulinism to counteract the high glucose levels, because swallowing a mucus-rich saliva with the meals delays gastric emptying (12).This paper is an attempt to test the truth of this supposition.     

Molecular characterisation of an Italian G6PD variant responsible for chronic non-spherocytic haemolytic anaemia

An Italian deficient G6PD variant associated with chronic non-spherocytic haemolytic anaemia (CNSHA) was biochemically characterised and studied at molecular level. Single-strand conformation polymorphism (SSCP) analysis led to the identification of an abnormal migration pattern of an amplified fragment encompassing exons 10 and 11 of the G6PD gene. Sequence analysis of both strands using an automated fluorescent DNA sequencer revealed a GA transition at nt. position 1246 in exon 10. A CT substitution at nt. 1311 in exon 11 was also found, which has already been described as a silent mutation common in Caucasians. The 1246 GA mutation has been described only in a Japanese subject with CNSHA (G6PD Tokyo) not associated with the 1311^T polymorphism, suggesting that this mutation may have arisen independently in Europe and Asia.     

消化道恶性肿瘤伴癌性腹水患者腹腔化疗前后血清及腹水IL-6和sIL-6R水平变化的意义

目的：探讨腹腔化疗对癌性腹水患者血清及腹水IL-6及sIL-6R水平影响及其临床意义。方法：采用放免法测定病人腹腔化疗前、治疗后四周血清及腹水IL-6水平变化，用ELISA同时测定sIL-6R水平变化，并与正常对照组相对比。结果：消化道恶性肿瘤伴癌性腹水患者血清IL-6和sIL-6R水平较正常对照组明显升高，腹水IL-6和sIL-6R水平较血清IL-6和sIL-6R水平明显增高。腹腔化疗后血清IL-6和sIL-6R水平下降，腹水IL-6和sIL-6R水平变化与血清水平变化相平行。且血清及腹水IL-6及sIL-6R水平变化与腹腔化疗是否有效有密切关系。结论：监测癌性腹水患者血清及腹水IL-6及sIL-6R水平变化是判断及预测腹腔化疗是否有效的途径之一。