肺部疾病中痰巨噬细胞对结核菌素试验反应的临床研究

目的:测定肺结核、肺肿瘤及非特异性炎症患者注射结核菌素前后痰巨噬细胞的不同数量变化,由此提供一种新的鉴别诊断方法。方法:皮内注射结核菌素、收集注射前、后痰液,测定痰巨噬细胞在不同时间的数量变化。结果:肺结核病人在结核菌素试验后痰巨噬细胞明显增加,肺部肿瘤及非特异性炎症则无显著性变化。结论:肺部患者测定结核菌素前后痰巨噬细胞变化,可作为菌阴肺结核与肺部肿瘤及非特异性炎症鉴别诊断依据之一。     

Relationship between cellular ATP content and cellular functions of primary cultured rat hepatocytes in hypoxia

The importance of oxygen in maintaining the functional integrity of hepatocytes has been well established in a variety of experimental models, such as in vivo , perfused liver and isolated hepatocytes. However, one of the shortcomings of these systems is their short life span. Therefore, we have examined the effects of long-term hypoxia on cellular adenine nucleotide content and cellular functions, such as albumin production, urea production and DNA synthesis, in adult rat hepatocytes in primary culture. Hepatocytes were cultured at a density of 11 × 10⁴ and 5 × 10⁴ cells/0.18 mL per cm² for the study of albumin and urea production and DNA synthesis, respectively, at various oxygen tensions (20, 12, 8 and 5%) for 24 h. Cellular ATP content in cultured hepatocytes in hypoxia gradually declined, corresponding to the decrease in oxygen tension, and the cellular ATP level at 5% oxygen was approximately 20% of that at 20% oxygen. Albumin production also decreased in parallel with the decrease in cellular ATP content in cultured hepatocytes in hypoxia. However, even when cellular ATP content gradually declined corresponding with the decrease in oxygen tension in cultured hepatocytes in hypoxia, such as at 8 or 5% oxygen, urea production remained at a high level; in contrast, DNA synthesis was completely suppressed. These results suggest that the cellular ATP content decreases in cultured hepatocytes during long-term hypoxia in relation to oxygen tension and that the relationship between decreased ATP levels and liver function in cultured hepatocytes during hypoxia differs for albumin production, urea production and DNA synthesis.     

High level production of soluble single chain antibodies in small-scale Escherichia coli cultures

We have investigated the effect of growth and induction conditions on the production of soluble single-chain Fv antibody fragments in Escherichia coli under the control of wt lac promoter. The scFv was directed into the periplasmic space by a pelB leader sequence. Addition of sucrose to the medium gave a 15–25-fold increase in the yield of soluble scFv-phOx (3.0 mg/l) for bacterial shake-tube cultures and an increase of 80–150-fold (16.5 mg/l) for shake-flask cultures. Using flask culture in the presence of 0.4 M sucrose, a significant amount of scFv was released into the medium. We found that the scFv could be made to accumulate in the periplasm or be secreted into the medium by simply changing the incubation conditions and the concentration of the inducer. The ratio between soluble antibody fragments and insoluble scFv aggregates proved to be dependent on the strength of the promoter. Lowering the incubation temperature below 20°C had no effect on the yield of soluble antibody fragments in the periplasm, but they were no longer secreted into the medium. An example of high level production in shake-flask cultures and one-step purification by immobilized metal affinity chromatography (IMAC) is described for a soluble scFv specific for the T cell surface antigen CD3. The biological activity of the purified anti-CD3 scFv was demonstrated by flow cytometry. This method should be especially useful for the functional screening of a large number of clones in small-scale cultures.     

A Correlation Between the Permeability Characteristics of a Series of Peptides Using an in Vitro Cell Culture Model (Caco-2) and Those Using an in Situ Perfused Rat Ileum Model of the Intestinal Mucosa

In an attempt to establish an in vitro/in situ correlation of intestinal permeability data, the permeability coefficients (P _app) for a series of model peptides, which were determined using an in situ perfused rat ileum model, were compared to the permeability coefficients (P _mono) determined using an in vitro cell culture model (Caco-2). The model peptides, which were all blocked on the N-terminal (acetyl, Ac) and the C-terminal (amide, NH2) ends, consisted of D-phenylalanine (F) residues (e.g., AcFNH₂, AcFFNH₂, AcFFFNH₂). To alter the degree of hydrogen bonding potential, the nitrogens of the amide bonds were sequentially methylated [e.g., AcFF(Me)FNH₂, AcF(Me)F(Me)FNH₂, Ac(Me)F(Me)F(Me)FNH₂, Ac-(Me)F(Me)F(Me)FNH(Me)]. These peptides were shown not to be metabolized in the in situ perfused rat ileum system. The results of the transport experiments showed that there were poor correlations between the apparent permeability coefficients (P _app) determined in an in situ perfused rat ileum model and the octanol–water partition coefficients (r = 0.60) or the hydrogen bonding numbers (r = 0.63) of these peptides. However, good correlations were observed between the in situ P _app values for these peptides and their partition coefficients in heptane–ethylene glycol (r = 0.96) and the differences in their partition coefficients between octanol–water and isooctane–water (r = 0.86). These results suggest that lipophilicity may not be the major factor in determining the intestinal permeability of these peptides and that hydrogen bonding potential may be a major contributing factor. A good correlation (r = 0.94) was also observed between the P _app values determined for these peptides in the in situ perfused ileum model and those P _mono values determined in the in vitro cell culture model (Caco-2) (Conradi et al., Pharm. Res. 8:1453–1460, 1991). These results suggest that the permeability values determined in the Caco-2 cell culture model may be a good predictor of the intestinal permeability of peptides.     

HM增菌液对沙门氏菌属与志贺氏菌属增菌效果的观察

报道HM增菌液对150例临床诊断为肠炎患者大便标本的增菌效果,并与革兰氏阴性杆菌增菌培养液(GN增菌液)和亚硒酸钠增菌液(SF增菌液)进行对比观察。结果显示HM增菌液对沙门氏菌属与志贺氏菌属增菌效果均较好,检出率明显高于GN和SF增萌液(p<0.01和p<0.05),有较好的实用价值。     

Efficacy of ivermectin for chronic strongyloidiasis: two single doses given 2 weeks apart

We assessed the efficacy and adverse effects of ivermectin for the treatment of chronic strongyloidiasis. Fifty patients were treated with a single dose of ivermectin at approximately 200 μg/kg, and the dose was repeated 2 weeks later. The eradication rate was 96% (48 of 50 patients) at 2 weeks after the first dose and 98% at 2 weeks after the second dose (49 of 50 patients). In the latter cases, no recurrence was noted at the end of the 4-month post-treatment follow-up period. The gastrointestinal symptoms observed most frequently before the treatment were borborygmus, constipation, and diarrhea. These symptoms improved in the majority of patients after the treatment. Side effects, including nausea and vomiting, occurred in 1 patient (2%), but these were mild and transient. Although abnormal values in laboratory tests were observed in 4 patients (8%), they were mild, and no particular clinical care was required. Thus, ivermectin showed an excellent antihelminthic effect, with little toxicity. Our results indicate that a regimen of two single doses of 200 μg/kg ivermectin, given 2 weeks apart, is clinically suitable for the treatment of chronic strongyloidiasis. Received: April 25, 2001 / Accepted: November 16, 2001     

60Coγ射线对体外培养成骨细胞作用的实验研究

目的：研究放射性骨损伤的发病机制，探讨骨折合并射线损伤愈合能力下降的原因。方法：将体外培养的成骨细胞用^60Coγ射线一次性照射3、6、9、12Gy，以建立体外培养成骨细胞受射线作用的实验模型；同时，观察成骨细胞的碱性磷酸酶（ALP）活性改变，骨形态发生蛋白（BMP）和转化生长因子β1（TGF-β1）的胞质内表达量，以及细胞内钙离子的含量。结果：成骨细胞受射线照射后，其ALP活性明显降低，且与照射剂量呈明显量效关系，3Gy组抑制率为31.59%，而12Gy组则为84.95%；BMP、TGF-β1的表达量下降，但与照射剂量无明显量效关系；胞质内钙含量亦降低，但受照各组间的抑制率无明显统计学差异。结论：射线对成骨细胞具有直接损害作用，放射性骨损伤的重要方面就是射线对成骨细胞的主要功能酶ALP活性的损害，抑制BMP、TGF-β1等生长因子的合成，且干扰细胞内第二信使钙离子，从而致骨组织的再生重建机能下降。     

大鼠小肠上皮细胞的体外原代培养

目的：探讨大鼠小肠上皮细胞原代培养的原理与方法以及培养过程中常见的问题和克服办法。方法：取生后1周SD大鼠的小肠上皮细胞，经分离纯化后进行原代培养。结果与结论：胶原酶Ⅺ型和中性蛋白酶Ⅰ型配合使用，可以分离到形态完整、容易贴壁生长的小肠绒毛细胞团。进一步利用离心技术纯化得到95％以上的小肠上皮细胞。结果显示，常规体外培养的肠上皮细胞是否增殖分化最终决定于用酶消化后的结构完整性，如果用0．25％胰酶消化，则细胞受损程度较大，不易贴壁生长。     

Splenocyte response to T cell-derived cytokines in thymectomized Xenopus.

A miniaturized, “hanging-drop” bioassay reveals that splenocytes from earlythymectomized (Tx) Xenopus can respond (by enhanced thymidine incorporation) to thymicdependent “cytokines” generated in PHA- or alloantigen-stimulated cultures. Preliminary evidence, using fluorescence activated cell sorting, indicates that surface IgM⁻ splenocytes, rather than sIgM⁺ cells, from Tx toads are sensitive to the crude, splenocyte-derived, active supernatants. Although these responsive cells display residual, but low, reactivity to PHA, their thymus independence is suggested by flow cytometric observations using the anti-T cell monoclonal antibody XT-1. The development of “T-like” cells in Tx Xenopus is discussed.     

微量沙眼衣原体组织培养法的建立及临床应用

建立了微量沙眼衣原体组织培养法，并应用于不同人群的生殖道标本的沙眼衣原体培养。结果生殖道沙眼衣原体培养的阳性率：男性尿道炎患者１４７％（９／６１）；正常孕妇３３％（７／２１０）；急性和亚急性盆腔炎女性患者６７％（７／１０４）；性病门诊患者１８２％（４９／２６９）。由于９６孔培养板及倒置荧光显微镜的应用，微量培养法试剂用量少、过程简化；自制的抗沙眼衣原体单克隆抗体大大降低了培养后的染色成本，适用于大宗标本。