Isolation of a cDNA clone for the liver cell adhesion molecule (L-CAM).

Liver cell adhesion molecule (L-CAM) is a calcium-dependent cell adhesion molecule found in very early vertebrate embryos and on liver and other epithelial cells in adults. To describe the genes coding for the molecule and study its synthesis, we have cloned cDNA from poly(A)+ RNA of 10-day embryonic chicken liver using the delta gt11 expression vector. One clone, lambda L301, has been characterized and used in analyses of L-CAM mRNA and genomic DNA. Clone lambda L301 produced a fusion protein that reacted strongly with polyclonal antibodies that recognize L-CAM (Mr 124,000) and its Mr 81,000 NH2-terminal fragment, Ft1, released from liver membranes by trypsin. This result indicates that lambda L301 contains a cDNA insert complementary to protein coding sequence within the two-thirds of the mRNA coding region beginning at the 5' end. The 220-base-pair cDNA insert was isolated and used as a probe in hybridization experiments. RNA transfer blot analysis of poly(A)+ RNA showed a single 4-kilobase mRNA; Southern blot analysis showed multiple components consistent with the presence of one to three L-CAM genes. To test whether different tissues express different forms of L-CAM message, poly(A)+ RNA from eight embryonic organs was analyzed. Only organs that expressed L-CAM protein contained poly(A)+ RNA that hybridized to the lambda L301 probe; in all cases a single band, with the same mobility as that in liver, was observed. The L-CAM mRNA in each tissue was present in proportions similar to those detected previously for the L-CAM protein in these tissues. The combined results suggest that any possible heterogeneity in the L-CAM genes is not reflected in the size of either the mRNA or protein.     

Rapid method for isolation of normal human peripheral blood eosinophils on discontinuous Percoll gradients and comparison with neutrophils

Previous studies on human eosinophils often have used cells from patients with hypereosinophilia syndrome or parasitosis owing to the difficulty in isolating pure populations of eosinophils from normal individuals. In the present study, human eosinophils were isolated with a purity of 97%, with 70% recovery from normal individuals with blood eosinophil counts of less than 3%. Human eosinophils are denser than neutrophils, but the range of densities of the two cell types overlap, making purification of eosinophils by density-gradient centrifugation difficult. However, if neutrophils were exposed to the chemotactic peptide (f-Met-Leu-Phe), which did not stimulate eosinophils, the neutrophils' density decreased, shifting them away from the density of eosinophils. Whole normal blood anticoagulated with EDTA was incubated at 37 degrees C for 15 minutes with 10(-6) mol/L f-Met-Leu-Phe and then layered over a discontinuous Percoll gradient (65% and 75% in diluted phosphate-buffered saline) and centrifuged at 400 g for 25 minutes at 22 degrees C. The cell layer between the 65% and 75% Percoll was collected and washed, and hypotonic lysis was used to remove erythrocytes. This cell layer contained 97.3 +/- 0.7% eosinophils (N = 8) with a yield of 4.9 X 10(4) eosinophils per milliliter of whole blood, or 70% of the total eosinophil count. The isolated eosinophils were in a quiescent state but responded to Escherichia coli endotoxin- activated serum with shape change and chemotaxis, membrane depolarization, and reduced nitroblue tetrazolium (96.0 +/- 1.0%), when stimulated with phorbol myristate acetate. In phagocytic assays, 89.3 +/- 1.3% of the eosinophils ingested Candida albicans v 96.0% +/- 1.0% of neutrophils. In contrast, the eosinophils did not respond chemotactically, alter membrane potential, or reduce nitroblue tetrazolium when treated with f-Met-Leu-Phe, and studies with f-Met-Leu- [3H]Phe showed that normal eosinophils lacked expression of receptors for f-Met-Leu-Phe. In control studies, normal eosinophils that were not exposed to f-Met-Leu-Phe during purification also failed to respond to f-Met-Leu-Phe, indicating intrinsic differences between normal eosinophils and neutrophils. Thus, exposure of whole blood to f-Met-Leu- Phe, followed by separation on Percoll is a simple method for rapid isolation of normal human eosinophils.     

表达白细胞介素10的系膜细胞基因转移载体的建立

目的为特异性地将病毒性白细胞介素１０（ｖＩＬ１０）基因转移到肾脏提供一种手段。方法应用逆转录病毒载体介导的基因转移技术将外源性基因ｖＩＬ１０转移到大鼠肾脏系膜细胞,应用ＲＴＰＣＲ、ＥＬＩＳＡ方法检测其表达,并通过对混合淋巴细胞反应（ＭＬＲ）的影响对其活性进行分析。结果ｖＩＬ１０在转染的系膜细胞表达２００００ｐｇ·（１０６细胞）－１／２４ｈ,应用３ＨＴｄＲ掺入法检测ｖＩＬ１０对ＭＬＲ的影响,实验组明显低于对照组（Ｐ＜００５）。结论外源性基因在系膜细胞有效地转录和表达,并对ＭＬＲ有抑制作用。     

产蓝色素放线菌的初步鉴定和蓝色素性质研究

对一株从土壤中筛选出来的产蓝色素的放线菌进行了初步鉴定, 命名为天蓝色链霉菌－１００（Ｓｔｒｅｐｔｏｍ ｙｃｅｓ ｃｏｅｌｉｃｏｌｏｒ－１００ ）,并对蓝色素的性质进行了研究．结果表明此色素无臭无味,水溶性较强．酸性条件下,溶解度随ｐＨ 降低而减小．ｐＨ＜ ８ 时为红色,ｐＨ≥８ 时为蓝色,颜色随ｐＨ 增大而加深．于１００ ℃短时间处理,比较稳定．在ｐＨ 中性的条件下对光（包括紫外光）较稳定．大多数金属离子如Ｂａ２＋ 、Ｋ＋ 等对此色素无影响,Ｍｇ２＋ 使色素增色．Ｆｅ２＋ 在５×１０－４ ｍ ｏｌ／Ｌ时,使色素变性,但在５×１０－５ ｍ ｏｌ／Ｌ时,对色素没有多大影响．Ｐｂ２＋ 可以络合色素,使之沉淀;加入０．２５％ 盐酸乙醇后,色素再溶解．该蓝色素可作为天然色素或天然ｐＨ 指示剂开发．     

血小板激活因子对急性重型胰腺炎肺损伤作用的实验研究

目的　探讨血小板激活因子（ Ｐｌａｔｅｌｅｔ Ａｃｔｉｖａｔｉｎｇ Ｆａｃｔｏｒ, Ｐ Ａ Ｆ） 在急性胰腺炎肺损伤发生发展中的作用。 方法　１７ 只纯种 Ｂｅａｇｌｅ 狗分成三组：（１） 胰腺炎组（ Ｐ 组） ,用牛磺胆酸钠加胰蛋白酶主胰管逆行注射,复制急性胰腺炎模型;（２） 治疗组（ Ｔ 组） ,模型复制后５ 分钟,３ 小时分别静脉注射苦杏内酯 Ｂ（ Ｂ Ｎ５２０２１） ;（３） 对照组（ Ｃ 组） 。分别测定平均肺动脉压值,肺湿重系数,支气管肺泡灌洗液（ Ｂｒｏｎｃｈｉａｌ Ａｌｖｅｏｌａｒ Ｌａｖａｇｅ Ｆｌｕｉｄ , Ｂ Ａ Ｌ Ｆ） 内细胞总数和蛋白含量,肺血管外水量／ 无血干肺重（ Ｅｘｔｒａ Ｖａｓｃｕｌａｒ Ｌｕｎｇ Ｗａｔｅｒ ／ Ｂｌｏｏｄ Ｆｒｅｅ Ｄｒｙ Ｌｕｎｇ , Ｅ Ｖ Ｌ Ｗ／ Ｂ Ｆ Ｄ Ｌ） ,肺组织内 Ｐ Ａ Ｆ含量。 结果　平均肺动脉压值 Ｐ组明显升高,在４ 小时与 Ｃ组、 Ｔ组差异均有显著意义（ Ｐ＜ ００５） 。 Ｐ组肺系数〔（１１２ ±００６） ％ ,珋ｘ ±ｓ ,以下同）〕, Ｅ Ｖ Ｌ Ｗ／ Ｂ Ｆ Ｄ Ｌ（５５９ ±０５３） 及 Ｂ Ａ Ｌ Ｆ中细胞总数〔（２０３ ±００２） ×１０９／ Ｌ〕和蛋白含量〔（４４７ ±     

人精子顶体反应的检测及其与穿卵试验的相关性

选择正常生育男子（生育组）、输精管结扎再吻合（吻合组）、不育症患者（不育组）各２５例的精液进行人精子顶体蛋白酶及酸性磷酸酶测定,顶体组织化学三色染色和人精子穿透去透明带金黄仓鼠卵试验,比较不同生育条件下精子功能和生育能力的相关性。结果显示,精子获能后顶体反应率明显高于获能前（Ｐ＜０．０１）。生育组顶体反应率和穿透试验高于吻合组和不育组（Ｐ＜０．０１）。结果表明,精子穿卵试验结合精子顶体反应测定可作为客观评价人类精子受精能力的依据     

飞行员直立位前后心率变异时域与频域的变化

为了探讨体位变化对心血管自主神经调节机制及心率变异（ＨＲＶ）特征的影响，对６０名健康男性战斗机飞行员进行了直立位（９０°）前后短时（５ｍｉｎ）时域与频域ＨＲＶ指标相关性研究，结果显示：直立位前后短时ＨＲＶ指标（包括时域和频域）存在明显差异，某些短时ＨＲＶ时域与频域指标，如卧位，５ｍｉｎＲ－Ｒ间期均值（ＭＲＲ５）与高频（ＨＦ）呈明显正相关，与高频／低频（ＨＦ／ＬＦ）呈明显负相关；直立位；ＭＲＲ５与低     

大肠癌患者血清脂蛋白、血脂、癌胚抗原分析

目的：观察大肠癌患者术前术后血清脂白、血脂、癌胚抗原的变化。方法：对４６例大肠癌病人和４０例正常人检测血清ＴＣ、ＴＧ、ＨＤＬ－Ｃ、ＬＤＬ－Ｃ、ＣＥＡ术前术后的改变。结果：术前大肠癌血清ＴＣ和ＨＤＬ－Ｃ水平明显低于正常组,ＣＥＡ水平明显高于正常组,差异非常显著（Ｐ〈０．０１）;术后大肠癌血清ＴＣ、ＨＤＬ－Ｃ、ＣＥＡ水平与正常组对照有显著差异（Ｐ〈０．０５）,大肠癌恶性程度高则血清ＴＣ和ＨＤＬ－Ｃ水平     

旋髂浅动脉游离皮瓣的临床应用及供瓣区处理

目的 探讨旋髂浅动脉皮瓣游离移植的临床适应证及供瓣区的修复方法.方法 根据创面大小设计旋髂浅动脉皮瓣,并分别游离移植修复17例面颈部、足踝部及小腿等处创伤及畸形,对供瓣区无法直接拉拢缝合者,设计同侧脐旁岛状皮瓣转移进行修复.结果 17例旋髂浅动脉游离皮瓣,最大面积19 cm×14 cm,最小5 cm×3 cm,16例术后成活良好,1例皮瓣部分坏死,术后2周移植皮片修复.10例供瓣区直接缝合,7例行脐旁岛状皮瓣转移修复,皮瓣全部成活,术后经3个月至2年随访,腹部外形良好.结论 旋髂浅动脉皮瓣游离移植,对于面颈部及手足等处具有良好的修复效果,而同侧脐旁岛状皮瓣亦可使供瓣区得到良好的修复.     

肾癌术前肾动脉栓塞及化疗性栓塞的疗效评价

目的　对 19例肾癌术前肾动脉栓塞及化疗性栓塞技术的疗效进行评价。方法　术前单纯性栓塞组 8例 ,化疗性栓塞组 11例 ,对其手术过程和病理切片进行回顾性研究。结果　所有病例均成功地实施术前栓塞 ,栓塞后手术可见肿瘤血供基本中断 ;病理可见肿瘤坏死明显且与周围组织界限清楚。讨论　肾癌术前肾动脉栓塞及化疗性栓塞对减少围手术期的风险及预后均有积极的影响