血沉标本中人巨细胞病毒DNA的检出

用聚合酶链反应（ＰＣＲ）和地高辛标记探针（ｄｉｇ－ｐｒｏｂｅ）检测临床血沉标本中人巨细胞病毒（ＨＣＭＶ）ＤＮＡ。结果表明,人群血标本中ＨＣＭＶ－ＤＮＡ携带率较高;ＰＣＲ技术较ｄｉｇ－ｐｒｏｂｅ更敏感、快速、简便,二者检出ＨＣＭＶ－ＤＮＡ阳性率分别为８３．３％和６０．３％;ＨＣＭＶ－ＤＮＡ检出率、ＨＣＭＶ－ＩｇＭ检出率与血沉值高低之间无相关性。     

用PCR技术克隆细小病毒H－1部分非结构蛋白基因

应用ＰＣＲ技术定向克隆了细小病毒Ｈ－１的非结构蛋白（ＮＳ）部分基因片段。自行设计并合成了ＰＣＲ引物△Ｐ３和△Ｐ４,在两个引物中分别引入两个突变碱基,使扩增后的ＤＮＡ片段的两端含有限制性核酸内切酶ＨｉｎｄⅢ或ＢａｍＨＩ的酶切位点,经双酿切法把该ＤＮＡ片段重组到ｐＵＣ１１８质粒中。对插入片段的ＤＮＡ序列测定和分析结果证实该片段为Ｈ－１ＮＳ－１基因序列。以此重组质粒为探针,采用分子杂交的方法,分别测定了Ｈ－１及ＭＶＭＤＮＡ在细胞内的复制水平。这一基因的克隆为制备Ｈ－１的质量监测、Ｈ－１及ＭＶＭＮＳ－１蛋白抑瘤作用机理及其在肿瘤细胞及正常组织中的转录表达等研究奠定了基础。     

抗原捕获聚合酶链式反应（AC—PCR）直接分型检测单纯疱疹病毒

用抗单纯疱疹病毒（ＨＳＶ）型共同性ｇＣ和ｇＤ单克隆抗体（ＭｃＡｂ）,包被Ｅｐｐｅｎｄｏｒｆ管,捕捉ＨＳＶ,同时加入３个引物：一个是ＨＳＶ－１／ＨＳＶ－型共同性上游引物,另两个分别是ＨＳＶ－１和ＨＳＶ－２型特异性下游引物。借此建立了能直接分型检测ＨＳＶ的抗原捕获聚合酶链式反应（ＡＣ－ＰＣＲ）。ＨＳＶ－１的扩增产物为４７７ｂｐ,ＨＳＶ－２的为３９９ｂｐ,两型病毒经ＡＣ－ＰＣＲ扩增后产生分子量不同的ＤＮ     

原位多聚酶链反应在人巨细胞病毒诊断中的应用

逝年来人巨细胞病毒（ＨＣＭＶ）感染率高,检测方法进展人,有巨细胞包涵体细胞（ＣＣＩＣ）检测、病毒分离、免疫检测、原位杂交（ＩＳＨ）、多聚酶链反应（ＰＣＲ）等法。目前有人把ＩＳＨ与ＰＣＲ结合创立了原位多聚酶链反应法（ＩＳＰＣＲ）,此法更准确、更敏感特异,更有地ＨＣＭＶ感染的早期快速诊断。本文就ＩＳＰＣＲ的原理、程序、特征主临床应用作一综述。     

广东两株人免疫缺陷病毒的分离和鉴定

从我省两名经性途径感染艾滋病的病人中采血,分离外周血单核细胞（ＰＢＭＣｓ）,将其与正常人ＰＢＭＣｓ共培养分离人免疫缺陷病毒（ＨＩＶ）,３周后检测上清ＨＩＶ－１ｐ２４抗原（ＥＬＩＳＡ法）超过阈值。将共培养第四周细胞和上清分别感染Ｈ９细胞（Ｔ细胞淋巴瘤传代细胞）,一周后检测ＨＩＶ－１ ｐ２４怕,证明有病毒生长。用ＨＩＶ－１ ＥＮＶ基因引物的套式聚合酶链反应（Ｎｅｓｔｅｄ ＰＣＲ）证实,两株新分离的病毒     

湖北省不同地区汉坦病毒M片段的分析研究

参考汉坦病毒（ＨＶ）的基因文库软件设计Ｍ片段Ｇ－１区型特异性引物,以ＨＶ　７６／１１８、Ｈ－１１４、Ａ－９及Ｒ－｛２２｝株ＲＮＡ为阳性模板,建立ＨＶ的分型方法──逆转录一半巢式聚合酶链反应（ＲＴ－ｈｅｍｉｎｅｓｔｅｄ　ＰＣＲ）,对湖北省不同地区分离的３０株ＨＶ进行分　型。结果显示：（１）建立的ＲＴ－ｈｅｍｉｎｅｓｔｅｄ　ＰＣＲ分型方法对ＨＶ两型的代表株７６／１１８（ＨＴＮ）、Ａ－９（ＨＴＮ）、Ｈ－１１４（ＨＴＮ）及Ｒ－｛２２｝（ＳＥＯ）ＲＮＡ进行了特异性扩增,其大小与理论值一致;此　法只能检测ＨＶ　ＲＮＡ,不能检测其它病毒ＲＮＡ。（２）分离于湖北省不同地区的３０株ＨＶ的分型结果　为汉滩型２１株、汉城型９株,其中长江以南汉滩型１２株、汉城型２株;长江以北汉滩型９株、　汉城型７株。这表明建立的ＲＴ－ｈｅｍｉｎｅｓｔｅｄ　ＰＣＲ用于ＨＶ的检测特异性好;分析湖北省不同　地区分离的３０株ＨＶ,显示湖北省ＨＦＲＳ流行为混合疫区;且在湖北地区具有一定的地理聚集性。     

用显微切割技术和催化信号扩增法检测何杰金病瘤细胞（H／RS）中的 …

为明确何杰金病（Ｈｏｄｇｋｉｎ’ｓ ｄｉｓｅａｓｅ,ＨＤ）瘤细胞中是否存在人巨细胞病毒（ｈｕｍａｎ ｃｙｔｏｍｅｇａｌｏｖｉｒｕｓ,ＨＣＭＶ）感染,采用显微切割技术结合ＰＣＲ方法检测ＨＤ组织及其瘤细胞Ｈｏｄｇｋｉｎ／Ｒｅｅｄ－Ｓｔｅｒｎｂｅｒｇ（Ｈ／ＲＳ）中的ＨＣＭＶ核酸;采用免疫组织化学催化信号扩增（ｃａｔａｌｙｓｅｄ ｓｉｇｎａｌ ａｍｐｌｉｆｉｃａｔｉｏｎ,ＣＳＡ）法检测ＨＤ中ＨＣＭＶ的立即     

逆转录-聚合酶链反应技术诊断丙型肝炎病毒感染

本文采用逆转录－聚合酶链反应（ＲＴ-ＰＣＲ）技术对２１２例住院及门诊病人其中肝病患者９８例（慢性肝炎４３例、肝炎后肝硬化４７例、原发性肝细胞癌８例）进行ＨＣＶ－ＲＮＡ检测。结果９８例慢性肝病患者血清中ＨＣＶ-ＲＮＡ-ＰＣＲ阳性２７例（２７．６％）,１１４例非肝病患者血清中ＨＣＶ-ＲＮＡ-ＰＣＲ阳性９例（７．９％）,两组间差异非常显著（Ｐ（０．０１）,各种肝病患者的ＨＣＶ-ＲＮＡ-ＰＣＲ阳性率均高于非肝病组。６８例患者同时进行了ＨＣＶ-ＲＮＡ-ＰＣＲ检测和抗－ＨＣＶ检测,２５例抗－ＨＣＶ阳性的患者中ＨＣＶ－ＲＮＡ-ＰＣＲ,２１例阳性（８４％）,４３例抗－ＨＣＶ阴性的患者中ＨＣＶ-ＲＮＡ-ＰＣＲ,９例阳性（２０．１％）、有输血及血制品史者４８例,其中ＨＣＶ-ＲＮＡ-ＰＣＲ阳性１６例（３３．３％）,１６４倒无输血史者中ＨＣＶ-ＲＮＡ-ＰＣＲ阳性２０例（１２．２％）,两组间差异非常显著（Ｐ（０．０１）。结果表明：１．ＨＣＶ感染与慢性肝病有密切联系,说明ＨＣＶ感染是慢性肝炎、肝硬化、肝癌的致病因素;２．ＨＣＶ-ＰＣＲ法具有特异性好、灵敏度高、简便快速等特点,弥补了抗－ＨＣＶ检测的不足之处,是目前确定ＨＣＶ感染的主要手段;３．ＨＣＶ感染与输血关系密切,因此对献血员进行常规ＨＣＶ检测对预防由输血所致ＨＣＶ感染有着极其重要的临床意义。     

HFRSV内影像型抗独特型人单抗重链可变区基因克隆

利用反转录－ＰＣＲ方法,从分泌肾综合症出血热病毒（ＨＦＲＳＶ）内影像型抗独特型人单抗杂交瘤细胞系（Ｃ８）中,成功地克隆了抗ＨＦＲＳＶ１ｄ人单抗重链可变区基因,并将此基因重组入Ｍ１３噬菌体ＤＮＡ中,测定了此重链可变区基因全序列。经计算机分析,基因全长共３５１ｂｐ、编码１１７个氨基酸,氨基酸序列同源性分析发现所推得的该单抗ＣＤＲ２区与ＨＦＲＳＶＧ２蛋白Ｃ端有同源区,此区可能具有较强的抗原性。     

应用反转录—聚合酶链反应检测口蹄疫病毒

建立了一种适用于检测动物（猪、牛、羊）组织（肌肉、淋巴结、脊髓、扁桃体和蹄冠皮）和牛食道－咽部分泌物（Ｏ－Ｐ液）中的口蹄疫（ＦＭＤ）病毒核酸（ＲＮＡ）的反转录－聚合酶链反应（ＲＴ－ＰＣＲ）技术。引物对是人工合成的两条２０ｍｅｒ寡核苷酸片段,它们的序列相应于ＦＭＤ病毒结构蛋白ＶＰ１基因后２／３区段,在４个血清型之间基本一致（保守）。ＰＣＲ产物经琼脂糖凝胶电泳检测。试验结果表明,ＲＴ－ＰＣＲ具有良好的     

TURNOVER OF PHOSPHATIDYLCHOLINE IN MICROSOMES AND MYELIN IN BRAINS OF YOUNG AND ADULT RATS

We have tested the hypothesis that the turnover of phosphatidylcholine in subcellular fractions of rat brain is a function of the age at which this lipid is deposited. Rats, 60 days of age, were injected intracranially with [2-³H]glycerol and either [methyl-¹⁴C]choline (to label the base moiety) or [U-¹⁴C]glucose (to label acyl moieties). Littermates were killed up to 90 days after injection and brain microsomes and myelin isolated. Lipids were extracted and the phosphatidylcholine was isolated by 2-dimensional TLC and hydrolyzed to its constituent moieties. The ³H in the glycerol backbone and ¹⁴C in the choline or acyl residues was quantitated. The microsomal and myelin ³H/¹⁴C ratios decreased with time with either set of precursors, indicating that labeled choline and acyl moieties were reutilized more efficiently than the glycerol backbone. The various precursors exhibited first order decay curves with half-lives for the glycerol backbone of 6 and 11 days for the microsomal and myelin fractions respectively. These results contrast with those previously obtained with identical experimental procedures when 17-day-old animals were injected. In that study, although much of the phosphatidylcholine turned over rapidly as for the older animals, by 2 weeks after injection most of the remaining phosphatidylcholine was turning over more slowly with a half-life of 13 and 25 days for microsomes and myelin respectively (Miller et al., 1977). The base and acyl moieties also had a corresponding shorter half-life in older animals relative to the slow turnover phase in younger rats.     

合成洗涤剂对人和哺乳动物细胞的诱变性研究

各种合成洗涤剂(洗衣粉,洗发膏,餐具洗涤剂等)产量日增。在日常生活中使用越来越普遍。洗涤剂直接或间接通过环境污染对人类健康产生影响,特别是潜在的致突变性引起公众的普遍注意。而现有的研究结果并不一致。本研究选用三种型号的合成洗涤剂,以小鼠生殖细胞染色体畸变和骨髓细胞微核率及离体的人类细胞和中国仓鼠细胞的染色体畸变和姐妹染色单体交换(SCE)为测定指标,系统地对合成洗涤剂的诱变活     

大鼠颌下腺促性腺激素释放激素及其mRNA的免疫组织化学与原位杂交研究

用免疫组织化学及原位杂交法,研究了促性腺激素释放激素及其ｍＲＮＡ在大鼠颌下腺的分布。结果显示,大鼠颌下腺的浆液性腺泡的上皮细胞,各级导管的上皮细胞及副交感神经节细胞均呈促性腺激素释放激素免疫反应阳性,阳性反应物质分布在胞质,胞核呈阴性反应。颌下腺的浆液性腺泡上皮细胞,各级导管上皮细胞同样被检测到很强的促性腺激素释放激素ｍＲＮＡ杂交信号。以上结果提示,大鼠颌下腺能自身合成促性腺激素释放激素,促性腺激素释放激素对消化功能可能有重要调节作用。     

武汉东湖沉积物和沉积物间隙水中氮和磷的含量及其分布

武汉东湖的沉积物中氮(N)的平均含量上层(0—10厘米)为14．2毫克／克(干重,下同),下层(10-40厘米)为12．0毫克／克。磷(P)的平均含量上层为0．87毫克／克,下层为0.62毫克／克。沉积物中氮和磷的含量均有明显的垂直和平面分布上的差异,上层高于下层,水果湖区高于其它湖区。沉积物间隙水中总溶解氮的浓度高,平均为儿．52毫克／升,主要以氨氮的形态存在,占总溶解氮的61．3％。总溶解磷的浓度较低,平均为0．113毫克／升,其中正磷酸盐占55.8％。间隙水中氮和磷的浓度均有明显的季节变动,夏季和秋季的浓度高,冬季和春季浓度低。除氨氮外,间隙水中氮和磷的浓度比底层湖水中氮和磷的浓度高,但一般不超过5倍,表明沉积物和湖水之间溶解的营养物质交换十分强烈。     

DEVELOPMENTAL CHANGES IN LEVELS AND FORMS OF CHOLINESTERASES IN MUSCLES OF NORMAL AND DYSTROPHIC CHICKENS

Abstract— Acetylcholinesterase (AChE) and pseudocholinesterase (°ChE) were studied in vivo and during the first several months of development of pectoral and posterior latissimi dorsi (PLD) muscles in normal and dystrophic chickens. Muscle extracts were prepared in a high ionic strength-nonionic detergent medium in the presence of protease inhibitors, in order to obtain complete solubilization and to prevent degradation of intrinsic molecular forms of both enzymes. In both normal and dystrophic pectoral muscles levels of AChE and °ChE increase rapidly in vivo, °ChE accounting for 5–10% of total cholinesterase activity. In the normal pectoral muscle the concentration of both enzymes drops rapidly after hatching with increasing muscle mass; total AChE per muscle remains relatively constant for 30 days post-hatch. In the dystrophic pectoral muscle both AChE and °ChE accumulate after hatching, resulting in greatly elevated levels (approx 10–25-fold) of both enzymes throughout the period studied. Multiple molecular forms of AChE and °ChE are observed in the pectoral muscle by sucrose gradient centrifugation. Four principal forms are distinguished: two light (L₁, L₂), one medium (M), and one heavy (H₂). The °ChE forms are 0.5–1.0 S units lighter than the corresponding AChE forms. L₂ is the predominant light form of AChE, whereas L₁ is the major light °ChE form detected. The lighter forms of AChE predominate in normal and dystrophic embryonic pectoral muscle at day 14, being replaced by the H₂ form by day 19. H₂ is the major °ChE form detected at day 19. After hatching, H₂ AChE is the predominant form found in both of the normal muscles studied. In the dystrophic pectoral muscle, progressive accumulation of the L₂ form of AChE is detected as early as day 4 post-hatch; this form eventually becomes predominant, although the heavier forms are also elevated. In PLD muscle the same phenomenon occurs, but with a slower time course. In dystrophic pectoral muscle a similar rise in the L₁ form of °ChE is first observed by day 4, with heavier forms also elevated in the mature muscle. Thus the alteration in the control of these two enzymes in dystrophic fast-twitch muscles results in an accumulation of the light forms of AChE and °ChE.     

VASCULAR SUPPLY AND STRUCTURE OF THE OVULE AND ARIL IN PEONY AND OF THE ARIL IN NUTMEG

Camp , Wendell H., and Mary M. Hubbard . (U. Connecticut, Storrs.) Vascular supply and structure of the ovule and aril in peony and of the aril in nutmeg. Amer. Jour. Bot. 50(2): 174–178. Illus. 1963.—Examination of the placental region in the carpel of Paeonia indicates a complexity and super-abundance of vascular supply beyond that usually found in angiosperms and certainly more than is necessary for adequate nutrition and water supply of the ovules. From this it is concluded that the ovules once were borne on a larger and more complex structure than the present carpel. Vascular strands leading to the aril and the hypostase are interpreted as being relictual. The large multifid aril of Myristica has a well-developed vascular system composed of several sizes of branched bundles.     

CORRELATION OF OXIDATION AND PHOSPHORYLATION IN HEMOLYZED BLOOD IN PRESENCE OF METHYLENE BLUE AND PYOCYANINE

1. The system:hemolyzed blood + glucose never exhibits glycolysis or, in the air, oxidation of glucose. When glucose is replaced by hexosephosphate ester, addition of methylene blue causes oxidation in air. 2. When cozymase is added also, the oxidation is increased, and a synthesis of hexosephosphate esters takes place. 3. When pyocyanine is used instead of methylene blue, the rate of oxidation is the same as with methylene blue, but a synthesis of phosphate esters takes place without addition of cozymase. 4. There is never a phosphate ester synthesis without oxidation going on, but oxidation does not necessarily go hand in hand with phosphate synthesis. 5. In order to couple the oxidation process with phosphate synthesis, two methods are available: either to start oxidation by methylene blue and to add coenzyme from yeast cells; or to start oxidation by pyocyanine, in which case coenzyme is unnecessary, though it improves the effect. 6. Iodoacetate always suppresses synthesis, but only under certain conditions decreases oxidation. Cyanide has no effect upon either process.