HBsAg重组DNA转化中国地鼠卵巢细胞二氢叶酸还原酶阴性突变株的细胞染色体变化及致瘤性

整合了含乙型肝炎病毒表面抗原(HBsAg)基因和dhfr基因的CHO-dhfr~-细胞,其染色体的畸变率和畸变类型都比亲代CHO-dhfr~-细胞高。但转化前后两系细胞的重要特性都未发生变异,即两者的染色体总数无差别,都是20条。两系细胞株接种裸鼠,均未发现有致瘤性。     

糙叶败酱挥发油镇静作用的研究

本文观察了败酱科植物糙叶败酱(Patrinia scabra Bunge)根和根茎中制得的挥发油的镇静作用,井与黄花败酱挥发油做了比较。结果表明,此油灌胃给予数组小鼠,剂量0.45ml/kg,显示如下的作用:[1]能显著延长由于腹腔注射戊巴比妥钠(50mg/kg)引起的小鼠睡眠时间,但其作用强度弱于黄花败酱挥发油。(2)一次灌胃给予小鼠大剂量10.46g/kg的糙叶败酱挥发油,连续观察10天,动物外观正常,无一死亡,体重增加与对照组相似。     

巨细胞病毒中性红斑定量试验与高滴度病毒的制备

在巨细胞病毒(CMV)的研究中常需对病毒定量。CMV需低滴度传代,否则会产生没有感染性的缺损病毒颗粒;CMV的抗原性受其感染量的影响;检测CMV中和抗体或纯化病毒都需具备病毒空斑定量基础。另外,制备高感染滴度的无细胞病毒(游离病毒)是对CMV进行分子生物学研究的前提。本文建立了CMV微量板法中性红斑定量技术并比较了几种制备无细胞CMV的方法。     

A dominant Th epitope in influenza nucleoprotein. Analysis of the fine specificity and functional repertoire of T cells recognizing a single determinant.

The sequence 260-283 of the nucleoprotein (NP) of influenza A virus is an epitope recognized by virus-immune lymph node cells from CBA (H-2k), B6 (H-2b), and B10.S (H-2s) mice. Further analysis shows that there are at least two Th epitopes within this sequence: the one close to the N-terminal (p260-273) is recognized by T cells from CBA and B6 mice while that close to the carboxyl-terminal (p270-283) is a dominant Th determinant in B10.S mice. The fine specificity of the recognition of this epitope by NP-specific T cell clones is also studied. When B10.S mice were infected intranasally or i.v. with live influenza virus, or immunized by different ways with various Ag preparations, P270-283 persistently emerged as a dominant T cell epitope. Immunization of B10.S mice with peptide p270-283 induces T cells with different in vivo functions including class II-restricted cytotoxicity, cognate help for Ag-specific antibody synthesis and delayed type hypersensitivity. This may have important implications for the understanding of the differentiation and classification of subsets of CD4+ T cells. The corresponding sequence of the NP of an equine influenza virus, A/Eq/Prague/56, which has a substitution (leucine to proline) at position 283, was not recognized by the lymph node cells from mice primed with either A/Okuda or A/Eq/Prague. However, the peptide, p270-283(E), representing this sequence induced T cell responses to both human and equine viruses. The data are discussed with respect to the development of viral vaccines.     

Thymic stromal cell clone with nursing activity supports the growth and differentiation of murine CD4+8+ thymocytes in vitro

Thymic stromal cell clone, TNC-R3.1 cell, was established from spontaneous AKR/J mouse thymoma. TNC-R3.1 cell, which has the similar properties to thymic nurse cells, formed a unique complex with normal thymocyte subpopulations. Flow cytometry analysis demonstrated that CD4+8+ and CD4-8- immature thymocytes preferentially interacted with TNC-R3.1 stromal cell clone. CD4+8+ thymocytes, which interacted with TNC-R3.1 stromal cell clone, contained a higher proportion of large size and cycling T cells than did noninteracting CD4+8+ thymocytes. As is generally accepted, CD4+8+ thymocytes did not respond to any stimulation such as IL-2, anti-CD3 mAb (2C11), or IL-2 plus 2C11. However, culture of isolated CD4+8+ thymocytes on TNC-R3.1 stromal cell monolayer in the presence of suboptimal dose of IL-2 induced a significant cell growth. Moreover, the addition of 2C11 and IL-2 into this coculture system resulted in a dramatic increase of the proliferative response of thymocytes. Flow cytometry analysis showed the proliferating cells on TNC-R3.1, which originated from CD4+8+ thymocytes, were mostly TCR-alpha beta+ CD3+CD4-8+ T cells. These results provide in vitro evidence that CD4+8+ thymocytes are at an intermediate stage of T cell maturation and TNC-R3.1 stromal cell clone induces the growth and differentiation of CD4+8+ thymocytes into CD4-8+ T cells.     

Serine-palmitoyl transferase activity in cultured human keratinocytes

Sphingolipids comprise approximately 25% of the stratum corneum lipids and are considered critical constituents of the epidermal permeability barrier. Whether sphingoid base structures are synthesized in the epidermis or whether they are derived from circulating or dermal sources is not known. We report here the initial characterization of serine-palmitoyl transferase (EC 2.3.1.50; SPT), the rate-limiting enzyme in the synthesis of sphingolipids, from cultured human neonatal keratinocytes. Subcellular fractionation studies demonstrated that 79% of the total cellular SPT activity was associated with the microsomes. The specific activity of keratinocyte SPT was 270 +/- 20 pmol/min per mg of microsomal protein, a level significantly higher than activities reported in other tissues. Keratinocyte SPT showed an apparent Km for L-serine of 0.40 (+/- 0.04 mM, with an alkaline pH optimum (8.2 +/- 0.4). Keratinocyte SPT utilizes palmitoyl-CoA preferentially over other saturated or unsaturated acyl-CoA substrates; increasing acyl-CoA chain lengths above C16 by one or two carbons was less detrimental to activity than similar decrements in chain length. Finally, the mechanism-based inhibitors L-cycloserine and beta-chloro-L-alanine, demonstrated potent inhibition of keratinocyte SPT activity, with 50% inhibitory concentrations of approximately 3.0 and 25 microM, respectively. In summary, we have found that cultured human neonatal keratinocytes contain unusually high levels of serine-palmitoyl transferase activity, and that the substrate specificity of keratinocyte SPT may determine the base composition of epidermal sphingolipids.     

The biological effect of sodium butyrate (NaBT) on SGC-7901 cells

Changes of (Na+-K+)-ATPase activity, cAMP and fibronectin (FN) content and cell surface microvilli were studied cytochemically, immunocytochemically and scanning electron microscopically on human stomach Glandular carcinoma (SGC-7901) cells treated with NaBT(2.5 mM). It was found that NaBT not only inhibited cell growth but also remarkably decreased the activity of cell surface (Na+-K+)-ATPase of SGC-7901 cells. Note worthy was that, in comparison with the untreated tumor cells, the increase of the intensity of intracellular cAMP and FN immunofluorescence in NaBT-treated tumor cells was striking. Moreover, in contrast to untreated tumor cells, the cell surface of NaBT-treated tumor cells showed more smooth and fewer microvilli under SEM. That NaBT may induce differentiation of SGC-7901 cells through inhibition of (Na+-K+)-ATPase activity and modulation of cellular cAMP and FN content was discussed.     

MPTP与帕金森氏症

1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)是一种神经毒,在人和灵长类动物,它选择性损害黑质纹体系统多巴胺神经元,使多巴胺(DA)及其代谢产物降低,引起典型的帕金森氏样症状。MPTP类毒性物质有其构效关系,其毒性主要与在脑内形成1-甲基-4-苯吡啶(MPP~ )有关。正常存在于脑内的色氨衍生物2-N-甲基四氢β-卡啉(2M-THBC),其构造及作用均类似于MPTP样物质,由此可以推断,MPTP除作为部分帕金森氏症的直接诱发原因外,极有可能是揭示原因不明的原发性帕金森氏病发病机理的一条重要途径。     

鲢、鳙在天然条件下的摄食强度(Ⅱ)武汉东湖鲢、鳙周年摄食强度的研究

武汉东湖的鲢(Hypophthalmichthy molitrix)鳙(Aristichthys nobilis)在天然条件下摄食强度具季节性变化。摄食强度高峰处于夏季,低谷处于冬季。在实验条件下,按周年采样期间水温变化范围,测定鱼的肠管排空率。食物通过鱼肠管时间(Y_p—h)与水温(X_t—℃)的关系为: 鲢Y_p=270.63 X_t~(0.6408) 鳙Y_p=280.46 X_t~(0.6642) 根据修正后Bajkov公式(D=C (24.A)/n),估算鱼的日粮。鱼日粮(Y_D)与水温(X_t)关系为: 鲢Y_D=0.2683e~(0.1503X_t) 鳙Y_D=0.0075X_t~(2.2715) 计算鱼在天然条件下周年月粮及年粮。鲢、鳙对天然饵料年消耗量分别为18.924公斤及17.39公斤,饵料系数分别为18.02及13.38。     

High frequency differences between high and low voltage electrocorticograms in the ovine fetus

Dawes (1986) has stated that, "The difference between high and low voltage activity depends solely on the presence in the latter of higher amplitude oscillations with relatively low frequency superimposed on the low voltage components as shown by spectral analysis". We have tested the constancy of the high frequency section of the power spectrum of the electrocorticogram in 7 near-term sheep fetuses. Under sterile conditions we implanted biparietal electrodes in the dura and a ground lead subcutaneously on the scalp. Five days after the surgery, with the animal standing quietly in the laboratory, we acquired the fetal electrocorticograms. Data were acquired during several high and low voltage electrocorticographic cycles in each animal. Two hundred power spectra were obtained during high and low voltage fetal electrocortical activity and statistically analyzed by paired t-test to discern differences in power between the high and low voltage pattern at each frequency (n = 7). We found that at all frequencies between 4 and 12 cycles/s the high voltage electrocorticogram had significantly more power than the low voltage electrocorticogram (P less than 0.05). This is in accordance with the established literature. We also observed that from 17 through 24 cycles/s the low voltage electrocorticogram is significantly higher than the high voltage electrocorticogram (P less than 0.05). In this frequency range the power of the high voltage expressed as a percentage of the power of the low voltage were respectively, 80, 74, 71, 66, 64, 64, 67, 64. These differences are of considerable magnitude and may be physiologically important.