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。     

用免疫荧光单克隆抗体对脊髓灰质炎病毒抗原表位的分析

用间接免疫荧光与中和试验筛选出来的抗脊髓灰质炎2型与3型不同毒株的12个单克隆抗体,其中3个仅有免疫荧光活性,9个具有中和与免疫荧光活性。用免疫荧光活性的单克隆抗体进行试验,发现它们在识别特异性抗原表位方面与中和性单克隆抗体相似,显示出株特异的、几个毒株共同特异的或型特异的抗原表位。根据表位分布关系及特征,可以用来鉴别型内毒株的特征、毒株的抗原分析、抗原变异的研究以及疫苗相关病例的鉴别。所得结果与中和性单克抗隆抗体及T1-寡核苷酸指纹图谱分析一致。而免疫荧光单克隆抗体识别抗原表位的活性范围比中和性单克隆抗体更广。另外还发现某些兼有荧光与中和两活性的同一个单克隆抗体,用不同方法(IF与NT)进行试验时,与相同毒株出现不同表位反应,这点是值得引起注意需待进一步证实的重要问题。     

Single-step purification of pertussis toxin and its subunits by heat-treated fetuin-sepharose affinity chromatography

A general procedure for purifying biologically active pertussis toxin from Bordetella pertussis fermentation broth using affinity chromatography on heat-treated fetuin-Sepharose CL-4B is described. Diethanolamine is used as eluent in this single-step purification to prepare endotoxin-free pertussis toxin in good yield (70%) and high purity (greater than 95%). This one-step affinity chromatography procedure can be easily applied for large-scale preparation of pertussis toxin S1 subunit and its B-component. The affinity-purified S1 subunit is devoid of any of the histamine-sensitizing activity normally associated with pertussis toxin. The chromatographically purified pertussis toxin and its subunits retained their immunogenicity and could induce high levels of anti-toxin neutralizing antibodies.     

Human and mouse LSP1 genes code for highly conserved phosphoproteins

With use of the mouse LSP1 cDNA we isolated a human homologue of the mouse LSP1 gene from a human CTL cDNA library. The predicted protein sequence of human LSP1 is compared with the predicted mouse LSP1 protein sequence and regions of homology are identified in order to predict structural features of the LSP1 protein that might be important for its function. Both the human and mouse LSP1 proteins consist of two domains, an N-terminal acidic domain and a C-terminal basic domain. The C-terminal domains of the mouse and human LSP1 proteins are highly conserved and include several conserved, putative serine/threonine phosphorylation sites. Immunoprecipitation of LSP1 protein from 32P-orthophosphate-loaded cells show that both the mouse and human LSP1 proteins are phosphoproteins. The sequences of the putative Ca2(+)-binding sites present in the N-terminal domain of the mouse LSP1 protein are not conserved in the human LSP1 protein; however, a different Ca2(+)-binding site may exist in the human protein, indicating a functional conservation rather than a strict sequence conservation of the two proteins. The expression of the human LSP1 gene follows the same pattern as the expression of the mouse LSP1 gene. Southern analysis of human genomic DNA shows multiple LSP1-related fragments of varying intensity in contrast to the simple pattern found after similar analysis of mouse genomic DNA. By using different parts of the human LSP1 cDNA as a probe, we show that most of these multiple bands contain sequences homologous to the conserved C-terminal region of the LSP1 cDNA. This suggests that there are several LSP1-related genes present in the human genome.     

酪氨酸对人离体滋养层细胞孕酮与hCG分泌的影响

本文观察三种剂量(2×10~(-5)mol/L,2×10~(-4)mol/L和2×10~(-3)mol/L)的酪氮酸对离体培养的滋养层细胞孕酮及hCG分泌的影响,并对其抑制效应的机理作了初步探讨。实验结果表明,三种剂量的酪氨酸均可抑制滋养层细胞孕酮分泌(P<0.01),但是,在孕酮分泌受酪氨酸抑制的同时,未见对hCG分泌发生影响(P>0.05),进一步观察了酪氨酸对滋养层细胞3β-羟甾脱氢酶活性的影响,结果表明,酪氨酸能显著抑制3β-羟甾脱氢酶活性,提示酪氨酸对滋养层细胞孕酮生成的抑制作用与抑制3β-羟甾脱氢酶活性有关。     

Cytokinin biochemistry in relation to leaf senescence. III. The senescence-retarding activity and metabolism of 9-substituted 6-benzylaminopurines in soybean leaves

A group of 14 9-substituted derivatives of 6-benzylaminopurine (BA), including the alanine conjugate, oxygen heterocyclic and alkyl derivatives, and compounds with a modified 9-ribose moiety, were assessed for their ability to retard soybean leaf senescence. The 9-alanine conjugate was very weakly active, and only two compounds, 9-(2-tetrahydropyranyl)-BA (9THP-BA) and 9-(2-tetrahydrofuranyl)-BA (9THF-BA), proved to be considerably more effective than BA. The metabolism of these three BA derivatives was determined to rationalize their differing activity. The alanine conjugate of BA was largely unmetabolized in leaf discs, but 9THP-BA and 9THF-BA released free BA and were also debenzylated to 9THP-adenine and 9THF-adenine, respectively. The three products of metabolism were identified by mass spectrometry. The enhanced activity of 9THP-BA and 9THF-BA, relative to that of BA, is attributed to their greater stability and their ability to gradually release free BA. This released BA was less susceptible to inactivation by alanine conjugate formation than was exogenous BA. The novel BA analogue 7-benzylaminooxazolo[5,4-d]pyrimidine, in which the 9-NH is replaced by oxygen, was inactive at 100 μM. For part II, see Zhang et al. 1987     

下肢长骨骨重的非对称性

本文测量82副中国成年下股长骨即股骨,胫骨和腓骨的重量,在两侧骨重相差>1％时,则股骨、胫骨和腓骨及其三骨总重均以非对称性为多,均具有非常显著性差异(P<0.001)。除腓骨重具有明显侧别差异外,其余无显著性侧别差异,对腓骨侧差解释为:①腓骨主要机能是肌的附着,而下肢对踢和足趾取物具有明显的侧别差异;②以两侧骨重相差>1％时为非对称性的标准,由于腓骨绝对重量较小,在两侧相差<0.5克时即多属非对称性。     

成人及新生儿颅骨铜和锌含量的初步研究

本文系用原子吸收光谱法测定了西安地区10例成人尸体和15例新生儿尸体的颅盖骨内Cu和zn的含量。结果求得成人颅骨中Cu正常含量(均数±标准差)为4.48±3.78mg/kg(干组织重);zn为597.05±472.54mg/kg。新生儿颅骨中Cu和zn的含量分别为1.96±0.76mg/kg;1160.38±859.71mg/kg。结果表明,成人颅骨内Cu含量高于新生儿,而成人颅骨内Zn含量显著低于新生儿。     

Detection of amyloid beta protein precursor immunoreactivity in normal and Alzheimer's disease cerebrospinal fluid

The amyloid A4 (or beta protein), a 4.2 kD polypeptide, is a major component of amyloid deposits in the brains of patients with Alzheimer's Disease (AD). The self-aggregating amyloid A4 protein of AD is encoded as part of three larger proteins by the amyloid A4 precursor gene. The corresponding proteins have 695, 751 and 770 amino acid residues. To investigate the utility of amyloid beta protein precursor (A beta PP) as a diagnostic marker for AD an antiserum against a synthetic peptide (175-186), predicted from cDNA sequence for A beta PP, was used. The immunoreactivity of A beta PP in normal and AD cerebrospinal fluid (CSF) was measured by Western blot and detected with radiolabeled protein A. A total of fifty-seven CSF samples (AD = 27 and normal = 30) were analyzed for A beta PP immunoreactivity. A polyclonal antibody detected two major protein bands with apparent molecular weights of 105kD and 90kD both in normal and AD CSF. The difference between normal and AD CSF was not significant. These results indicate that immunoreactivity of A beta PP is present both in normal and AD CSF, and that the difference is too small to be used as a diagnostic marker.     

Morphological and molecular characterization of fertile tetraploid somatic hybrids produced by protoplast electrofusion and PEG-induced fusion between Lycopersicon esculentum Mill. and Lycopersicon peruvianum Mill.

Summary Mesophyl protoplasts of two genotypes of cultivated tomato (Lycopersicon esculentum Mill.) and one of its wild relative species (Lycopersicon peruvianum Mill.) were fused by using electrofusion and polyethyleneglycol-induced fusion. Forty-three fertile tetraploid somatic hybrid plants, each deriving from separate calli, were recovered from both fusion procedures. Electrofusion appeared more efficient than chemical fusion for the production of somatic hybrids. These plants appeared morphologically similar, whatever the fusion procedure and tomato genotype. They had intermediate leaf, inflorescence, and flower morphology. After self-pollination, the hybrids set fruit of intermediate size and color. The hybrid nature of these plants was confirmed by isoelectric focusing of the Rubisco small subunits used as nuclear markers. L. esculentum and L. peruvianum were distinguished by means of two chloroplast markers: CF1-ATPase subunit as analyzed by isoelectro-focusing and ct DNA restriction patterns. All hybrids displayed both ct markers of only one parent with no biased transmission. Mitochondrial (mt) DNAs were prepared from flower buds by using miniaturized CsCl gradients. Preliminary analysis indicated that mt genomes from the hybrids all differed from those of both parents. mt DNA Sall restriction enzyme analysis revealed that all but two hybrids contained one novel fragment of 13.5 kb. Gene mapping experiments showed that the mt apocytochrome b and ATPase subunit 9 homologies in the somatic hybrid mt DNA resembled L. esculentum and L. peruvianum, respectively; the mt nad5 probe distinguished at least four distinct patterns in the hybrids. These results indicated that mt DNA rearrangements involving intergenomic recombinations occurred through protoplast fusion. A greater mt DNA polymorphism was induced with chemical fusion than with electrofusion.