HBcAg显性失活突变体质粒的构建及体外表达

目的：探讨HBcAg显性失活突变体质粒pCDNA4-C-GFP的构建及体外表达。方法：选择编码乙肝核心抗原C基因片段及绿色荧光蛋白（green fluorescent protein,GFP)基因片段，利用基因重组技术构建成DNA质粒pCDNA4-C-GFP,并将该质粒转染肝癌细胞株HepG2。结果：通过RT－PCR检测到其RNA的表达，Confocal观察到GFP绿色荧光。结论：HBcAg显性失活突变体质粒pCDNA4-C-GFP的构建成功可以进行对HBV作用的研究。     

骨感染病人TNF-α、IL-6和C反应蛋白的检测及临床意义

①目的　探讨骨感染病人肿瘤坏死因子α(TNF α)、白细胞介素 6 (IL 6 )和C反应蛋白 (CRP)水平的变化及意义。②方法　采用双抗体夹心ELISA法测定 30例骨感染病人及 2 5例正常人的血清TNF α、IL 6水平 ,采用散射比浊法测定CRP水平。③结果　骨感染病人血清中TNF α、IL 6及CRP含量均处于高水平状态 ,与健康人比较 ,差异有显著性 (t=2 .2 6～ 10 .6 7,P <0 .0 5、0 .0 1)。④结论　TNF α、IL 6及CRP是预测骨感染病人病情危重的重要指标之一 ,对临床有一定指导意义     

Purification of rabbit, rat and mouse protein C with the use of monoclonal antibody to human protein C, PC01

Ca(++)-dependent monoclonal antibody specific to gamma-carboxyglutamic acid (Gla) domain of protein C was produced. It did not cross-react to the other vitamin K-dependent plasma proteins but to protein C of the other species. Using this monoclonal antibody, PC01, rabbit (170 micrograms), rat (60 micrograms) and mouse (40 micrograms) protein Cs were isolated from 100 ml of their plasma by affinity chromatography. All of these protein Cs were two chain form linked by disulfide bond as well as human protein C and activated by thrombin-thrombomodulin complex. Rat and mouse protein Cs showed similar characters to human protein C. On the other hand rabbit protein C had different M(r) of heavy and light chains and showed lower anticoagulant activity compared with human protein C.     

Developmental patterns of intermediate filament gene expression in the normal hamster brain

We have examined the patterns of expression of the major intermediate filament (IF) protein mRNAs during development of the hamster brain. Quantitative northern blotting was used to examine changes in the levels of mRNAs for the low, middle and high molecular weight neurofilament proteins (NF-L, NF-M, NF-H) as well as peripherin, vimentin and glial fibrillary acidic protein (GFAP). Total RNA was isolated from hamster brains at embryonic (E) days 12 and 14 and postnatal (P) days 1, 3, 5, 7, 9, 11, 13, 15, 20, 28 and 60-90 (adult), and probed with specific IF cDNAs. Northern blotting revealed that NF-L and NF-M mRNAs were present at very low levels in embryonic brain and that significant expression of these genes only occurred postnatally when the levels increased dramatically until P28 and then declined again in the adult. Increases in NF-H mRNA levels were somewhat delayed relative to those of NF-L and NF-M. NF-H mRNA was not seen at embryonic stages and was expressed at very low levels prior to P9; after that time the levels increased rapidly until P28 and then declined in the adult. Two of the type III IF genes, peripherin and vimentin, followed a pattern of expression opposite that of the NF genes. Both peripherin and vimentin mRNAs were present in embryonic brain and were expressed at higher levels during early postnatal stages than at later times. The magnitude and rate of reduction in vimentin gene expression in the postnatal interval was much greater than that of peripherin. GFAP mRNA levels were extremely low prior to P9 after which a robust increase occurred, followed by a decline in the adult. We discuss the implication of the dramatic changes in IF isotype expression in brain to the pathways of both neuronal and glial development in vivo.     

Distribution of cytochrome oxidase and parvalbumin in the primary visual cortex of the adult and neonate monkey,Callithrix jacchus

The anatomical distributions of the mitochondrial enzyme cytochrome oxidase (CO) and of the calcium binding protein parvalbumin (PV) were studied in the striate cortex of adult and neonate New World monkeys (Callithrix jacchus). In the adult marmoset, both proteins were found in laminar arrangements similar to those described for the macaque monkey, with prominent bands of PV-like immunoreactive (PV-LI) puncta in layers IV and IIIb, and fairly evenly distributed PV-LI nonpyramidal neurons. Furthermore, the pattern of CO activity in area 17 of the neonate marmoset was almost identical to the CO pattern described in neonate macaque and squirrel monkeys. It came, therefore, as a surprise to find that the adult pattern of PV-like immunoreactivity (PV-LI) in the marmoset striate cortex arises from a neonatal pattern strikingly different from that seen in any developmental stage of the macaque, or in any other mammal studied so far. In the deep layers IV through VI of the neonate marmoset, a large number of PV-LI neurons was stained in bandlike patterns, their number in layers IV and V exceeding the number of PV-LI neurons present in these layers of the adult marmoset area 17. Staining of layers IV and VI was restricted to area 17 and involved nonpyramidal cells and their exceeding the number of PV-LI neurons present in these layers of the adult marmoset area 17. Staining of layers IV and VI was restricted to area 17 and involved nonpyramidal cells and their processes. The stained band of layer V, in contrast, continued throughout most of the neocortex. In area 17, an estimated 10 to 20% of the stained cells in layer V exhibited pyramidal shapes. The findings show that the expression of PV by visual cortical cells occurs before birth and suggest that the comparatively early onset of PV expression is not dependent on the onset of textured vision. The exuberant number of stained cells in some layers, and particularly the staining of pyramidal cells, in the neonate marmoset, suggest that a considerable number of cells possesses the stainability for PV-LI only transiently, i.e., in the marmoset, these cells have a specific demand for parvalbumin during this phase of their development. © 1994 Wiley-Liss, Inc.     

Atypical Allergic Colitis in Preterm Infants

ABSTRACT. Two atypical cases of colitis due to cow's milk protein intolerance (CMPI) are reported, affecting preterm infants. One developed a toxic dilatation of the colon and responded well to a casein hydrolysate based feed. The second presented insidiously and failed to tolerate a casein hydrolysate, but responded well to a chicken-based modular feed.     

Chronic ethanol intake may delay the onset of gossypol-induced infertility in the male rat

Summary. Parameters were obtained from the reproductive organs of ethanol-fed, gossypol-treated Sprague Dawley rats. The experimental animals were fed either on a normal (15.17%) or low protein (8.00%) diet. Measurements included reproductive organ weights, seminal characteristics, serum concentration of testosterone and histological, stereological and histomorphometric evaluation of the testis. The testis size, length and diameter of the seminiferous tubule had the least values in the protein-malnourished, gossy-pol-treated rats (3.01±0.26 g, 0.56 ± 0.03 m, 281.34±11.30 μn), in comparison to corresponding animals which had received ethanol simultaneously with gossypol (3.40 ± 0.25, 0.71±0.06m, 314.42 ± 11.61 μn). As gossypol and ethanol are both associated with reduced reproductive capacity, this unexpected but interesting finding lends support to the hypothesis that either a normal dietary protein or ethanol consumption may modify the action of gossypol on body tissues, including the testis. This effect, presumably mediated through changes caused to the bioavailability of gossypol, modifies its antifertility activity. The present observation further highlights the need to consider the concurrent administration of other drugs, such as alcohol, and the nutritional status in the evaluation of gossypol for various potential uses.     

Effect of dipyridamole with or without aspirin on urine protein excretion in patients with membranous glomerulonephritis

Summary The antiproteinuric effect of the antiplatelet agent dipyridamole has been assessed after inhibiton of thromboxane B₂ (TxB₂) synthesis in 8 patients with confirmed membranous glomerulonephritis. There were three study periods, each of 30 days, and 45 days apart, namely a washout period, treatment with dipyridamole 300 mg/d, and dipyridamole 225 mg/d plus aspirin 150 mg/d. On Days 1 and 30 of each study period serum and urine creatinine, 24-h excretion of protein, creatinine clearance, platelet aggregometry on whole blood and serum TxB₂ were measured. Treatment with dipyridamole alone or with aspirin produced significant inhibition of platelet aggregation and a fall in 24-h protein excretion; the latter amounted to 54% with dipyridamole alone and 56 % with dipyridamole plus aspirin (NS). Dipyridamole plus aspirin caused an 82 % reduction in serum TxB₂.     

脂多糖和同型半胱氨酸对人脐静脉内皮细胞单核细胞趋化蛋白1 mRNA表达的影响

目的探讨脂多糖和同型半胱氨酸是否诱导培养的人脐静脉内皮细胞表达单核细胞趋化蛋白1(MCP-1) mRNA及其机制.方法将生长至汇合的人脐静脉内皮细胞分为对照组、脂多糖组,脂多糖+SB203580组,同型半胱氨酸组,同型半胱氨酸+SB203580组,脂多糖+同型半胱氨酸组.采用斑点杂交和RT-PCR检测其MCP-1 mRNA的表达.用免疫细胞化学法检测对照组、脂多糖组和同型半胱氨酸组P38蛋白激酶蛋白的表达.结果培养的人脐静脉内皮细胞能表达较低水平的MCP-1 mRNA.斑点杂交和RT-PCR均显示各实验组的MCP-1 mRNA明显高于对照组.免疫细胞化学显示,P38蛋白激酶蛋白在对照组的细胞核阳性表达率为9.6%,当人脐静脉内皮细胞暴露于脂多糖或同型半胱氨酸后,细胞核阳性表达率升至46.7%或57.7%.结论脂多糖和同型半胱氨酸能诱导人脐静脉内皮细胞表达单核细胞趋化蛋白1 mRNA,P38蛋白激酶可能参与了该过程.