新生牛血清中乙脑抗体的检测及其方法的建立

对新生牛血清中的抗乙型脑炎病毒的抗体进行检测,并且建立一种有效可行的检测方法。以四个厂家共16批新生牛血清用蚀斑减少中和法(PRNT)对其中的乙脑抗体进行了检测。结果显示该方法检测出部分批次的新生牛血清中含有抗乙脑病毒抗体,且乙脑抗体阳性牛血清可将乙脑病毒抗原中和,对蚀斑数有较明显影响。证明作为检测牛血清中乙脑抗体的有效方法,PRNT法具有灵敏度高、重复性好的优点。以上检测说明新生牛血清中存在乙脑抗体,能将乙脑病毒中和。用于乙脑减毒活疫苗生产的新生牛血清除按照《中国生物制品主要原辅材料质控标准》进行质控外,还应进行乙脑抗体的检测。     

Transgenic models to study disorders of respiratory control in newborn mice

Recent studies described the in vivo respiratory phenotype of mutant newborn mice with targeted deletions of genes involved in respiratory control development. Whole-body flow barometric plethysmography is the noninvasive method of choice for studying unrestrained newborn mice. The main characteristics of the early postnatal development of respiratory control in mice are reviewed, including available data on breathing patterns and on hypoxic and hypercapnic ventilatory responses. Mice are very immature at birth, and their instable breathing is similar to that of preterm infants. Breathing pattern abnormalities with prolonged apneas occur in newborn mice that lack genes involved in the development of rhythmogenesis. Some mutant newborn mice have blunted hypoxic and hypercapnic ventilatory responses whereas others exhibit impairments in responses to hypoxia or hypercapnia. Furthermore, combined studies in mutant newborn mice and in humans have helped to provide pathogenic information on genetically determined developmental disorders of respiratory control in humans.     

The creation of specific immunity to staphylococcal infection in newborn infants by the intranasal administration of adsorbed staphylococcal anatoxin

The possibility of enhancing specific immunity in newborn infants by the intranasal administration of adsorbed staphylococcal toxoid to infants with a high risk of staphylococcal infection in doses of 1 drop (0.05 ml) into each nostril during the first 7-9 years of their life. On days 7-9 the level of anti-alpha-toxin in the blood rose to 3.8 +/- 0.14 I. U./ml and remained sufficiently high 3-6 months later. When this method was used for the simultaneous immunization of mothers, their antitoxic titers were not as high as in newborn infants. No side effects were observed. In the control group, the titers of anti-alpha-toxin were low during the whole period of observation. Infants immunized by the proposed method had no staphylococcal infections both during the newborn period and within the first year of their life. In the control group, 8 cases of minor forms of purulent septic infection were registered during the newborn period, and in 2 infants umbilical staphylococcal sepsis was diagnosed.     

Neural control of breathing: insights from genetic mouse models.

Recent studies described the in vivo ventilatory phenotype of mutant newborn mice with targeted deletions of genes involved in the organization and development of the respiratory-neuron network. Whole body flow barometric plethysmography is the noninvasive method of choice for studying unrestrained newborn mice. Breathing-pattern abnormalities with apneas occur in mutant newborn mice that lack genes involved in the development and modulation of rhythmogenesis. Studies of deficits in ventilatory responses to hypercapnia and/or hypoxia helped to identify genes involved in chemosensitivity to oxygen and carbon dioxide. Combined studies in mutant newborn mice and in humans have shed light on the pathogenesis of genetically determined respiratory-control abnormalities such as congenital central hypoventilation syndrome, Rett syndrome, and Prader-Willi syndrome. The development of mouse models has opened up the field of research into new treatments for respiratory-control disorders in humans.     

Absnece of amyloid protein in the organs of normal mice]

The immunomorphological Coons' method (its indirect variant) was used to discover specific amyloid protein AA in the organs of mice with experimental amyloidosis and normal mice (adult, newborn, embryo). The use of pure antiprotein-AA-antibodies allowed to reveal the minimal deposits of amyloid protein at the earliest stages of amyloidogenesis. The organs of normal mice (adult, newborn, embryo) did not contain this protein.     

Trichinella spiralis: modifications of the cuticle of the newborn larva during passage through the lung.

A scintigraphic method was developed to study the distribution of radioactivity after iv injection of 131I-labeled Trichinella spiralis newborn larvae into normal rats. It was found that the radioactivity was immediately retained in the lungs and thereafter slowly released, with a mean transit time in excess of 9 hr, as calculated by image analysis. At various times after iv injection of newborn larvae into normal mice, the lungs were removed and parasites were recovered and counted. Fifty to seventy percent of the larvae injected were recovered after 30 sec, between 10 and 30% after 1 min, and less than 4% at 15 min. These results indicate that during the very rapid passage of newborn larvae through the lungs, labeled components of the cuticle are detached and retained. It is suggested that the modifications produced in the cuticle of the newborn larva during its passage through the lung may increase its resistance to the nonspecific defense mechanisms of the host.     

IN VITRO BIOSYNTHESIS OF TUBULIN ON TOTAL, FREE AND MEMBRANE-BOUND POLYSOMES FROM THE DEVELOPING RAT BRAIN

Abstract— Incorporation of [³H]leucine into tubulin and total protein was examined using a polysomal system from newborn (1-day-old). young (10-day-old) and adult (3-month-old) rat brains and cerebral cortices. The rate of tubulin biosynthesis (specific radioactivity) was always lower than that of total protein biosynthesis. No significant differences in the specific radioactivities of the synthesized total proteins were found between the newborn and young brain polysomal system, although young cerebral cortical polysomes were less active than newborn cerebral cortical polysomes. The adult brain (or cerebral cortical) polysomes were less active, about 20-30% lower than the young brain (or cerebral cortical) polysomes. The incorporation of [³H]leucine into tubulin showed a progressive decrease in the polysomal systems isolated from the newborn, young and adult rat brains and cerebral cortices. These tendencies were similar in every cell sap taken from newborn, young and adult rat brain homogenates.
In order to examine the relative activities of free and bound polysomes of the developing rat brain in tubulin biosynthesis. double-labelling experiments were carried out. Labelled tubulin was purified by the assembly and disassembly method, followed by SDS gel electrophoresis, or by vinblastine precipitation method, followed by SDS gel electrophoresis; then identification by co-electrophoresis with native brain tubulin, molecular weight determination and demonstration of specific aggregation in the presence of GTP followed. Free and bound polysomes showed approximately similar activities during tubulin biosynthesis. Furthermore, relative activities of tubulin biosynthesis by free and bound polysomes did not significantly change during development.     

Method for the quantification of underivatized amino acids on dry blood spots from newborn screening by HPLC-ESI-MS/MS

In our study we have developed an HPLC-ESI-MS/MS method for qualitative and quantitative analysis of underivatized amino acids on dry blood spots. The sensitive and specific instrumental performances permitted the chromatographic separation of 40 amino acids and their isomers within 10 min. The method has been set up for cases of suspected metabolic diseases revealed by newborn screening. What is new is that it is applied on the same blood spots used for newborn screening, instead of plasma, in order to avoid involvement of doctors, increased anxiety for parents, stress for patients for plasma collection, long time of waiting and further costs for analysis.     

Measurement of pulmonary mechanics in the newborn lamb: a comparison of three techniques

Although recent interest in neonatal respiratory mechanics has led to the development of a plethora of techniques for measuring lung compliance and resistance, a critical appraisal of the limitations of these techniques in the newborn has not been performed to date. We evaluated three techniques of measuring respiratory mechanics in the newborn lamb, with the reference method (method 1) being the Mead-Whittenberger technique using flow, volume, and esophageal pressure (Pes) by water-filled catheter, and the other two methods entailing the measurement of mouth pressure (Pm) during airway occlusion (method 2 using end-expiratory occlusion; method 3 using end-inspiratory occlusion). Each technique was evaluated during eupnea and tachypnea in intubated and nonintubated newborn lambs. We found that the use of Pes for the measurement of resistance and compliance gave the most reliable results during both eupnea and tachypnea in both the intubated and nonintubated subjects. The airway occlusion techniques that use Pm to derive resistance and compliance (methods 2 and 3) gave more variable results under all conditions of testing. Method 2 was the least precise method of measurement with a variability of greater than 30% compared with a variation of less than 20% for method 1. For all three methods, it was found that the number of breaths needed for reproducible measurements of mechanics was four to six during eupnea and seven to nine during tachypnea.     

Fate of polyoma origin of replication after its direct introduction into mice

Recently we have developed a method for direct introduction of calcium phosphate-precipitated DNA into newborn rats. To examine whether the foreign DNA can replicate, a plasmid containing a polyoma origin of replication was injected into newborn mice. The plasmid was found intact in liver and spleen and able to transform bacteria. The foreign DNA had disappeared by the seventh day after injection. Yet, the plasmid DNA containing the polyoma origin of replication had undergone replication in both the liver and the spleen.     

In vivo adenovirus-mediated gene transfer to newborn rat retinal pigment epithelial cells

A successful surgical access to the subretinal space is critical for achieving adenovirus-mediated gene transfer to the retinal pigment epithelial (RPE) cells or photoreceptor cells. We report a novel surgical approach allowing an efficient delivery of recombinant replication-deficient adenoviral vectors into the subretinal space of newborn rats. Our data suggest that this method may be useful for infecting reproducibly large areas of the RPE cell layer of normal newborn rats and should be applicable to RCS pups. We also show the feasibility of infecting ex vivo RPE cells in culture using the same recombinant adenoviral vector.     

Cytochemical analysis of single villus peptidase activities in pig intestine during neonatal development

The present work uses a new technique of whole tissue cytochemistry and automated scanning to obtain measurements of peptidase activity and surface structure in intact villi microdissected from the jejunum of newborn and 28-day-old pigs. Intact villi from 28-day-old pigs are shown by this method to contain 30% more aminopeptidase N and 400% more dipeptidylpeptidase IV activity than is found on villi taken from newborn pig intestine. Villi taken from 28-day-old pig intestine are also half as long and twice as wide at their base as those taken from newborn animals. These changes in shape take place without significantly affecting the total surface area of the villus. Increases in peptidase activities occurring during postnatal development can be further subdivided into those dependent upon induced changes in enterocyte biochemistry and those dependent upon the changing geometry of villi. Over 90% of the total increase in peptidase activities occurring during neonatal development was shown, by this analysis, to involve enterocyte reprogramming of enzyme production. The present method of whole tissue cytochemistry appears to combine successfully the ability to measure peptidase activities at the cellular level in undisrupted tissue with a facility to relate these results to the overall shape of a single villus. These and more general applications of the method now provide new ways to analyse a variety of changes taking place in intestinal structure and function in a quantitative manner.     

Production of minimally disturbed synchronous cultures of hematopoietic cells

A method is describedforproducing sizable quantities of synchronously dividing, minimally disturbed mammalian cells. Cultures were grown immobilized on surfaces such that cell division within the population resulted in the continuous release of synchronous newborn cells. As judged by the quality and duration of synchronous growth, cell size distributions, and DNA compositions, newborn mouse L1210 cells grew with a very high level of synchrony without overt evidence of growth disturbances. The technology should be applicable to a variety of hematopoietic cells, as evidenced by similar results with human MOLT-4 and U937 cell lines.     

Determination of total insulin (TIRI) in plasma of insulin-treated diabetics and newborn infants of insulin-treated diabetic mothers.

Plasma of insulin-treated diabetics and of newborn infants of insulin-treated diabetic mothers contains insulin antibodies which invalidates the radioimmunoassay of insulin. Therefore, the endogenous insulin antibody complex must be splitted at a pH lower than 5 and the total IRI (TIRI) is separated by ethanol extraction. It was investigated the recovery rate in dependence upon plasma volume used for extraction. By reduction of used plasma volume from 500 to 200 mul per extraction the recovery rate was increased from 65.1 +/- 8.4 to 88.3 +/- 4.2% (mean +/- SEM). The low plasma volume of 200 mul for TIRI extraction made it possible to determine TIRI during glucose loads of newborn infants. To eliminate different conditions of incubation for standard and unknown plasma samples the TIRI levels were computed by means of so-called "extracted" standard curve, obtained with extracted insulin from standard insulin dilution in insulin-free pooled human plasma. Using the described method a temporary regeneration of insulin secretion of a newly diagnosed juvenile diabetic after insulin treatment could be shown. In contrast to newborn infants of healthy mothers a biphasic/insulin release was found during the intravenous glucose loads in newborn infants of insulin-treated diabetic mothers.     

Automatic classification of activity and apneas using whole body plethysmography in newborn mice.

An increasing number of studies in newborn mice are being performed to determine the mechanisms of sleep apnea, which is the hallmark of early breathing disorders. Whole body plethysmography is the method of choice, as it does not require immobilization, which affects behavioral states and breathing. However, activity inside the plethysmograph may disturb the respiratory signal. Visual classification of the respiratory signal into ventilatory activity, activity-related disturbances, or apneas is so time-consuming as to considerably hamper the phenotyping of large pup samples. We propose an automatic classification of activity based on respiratory disturbances and of apneas based on spectral analysis. This method was validated in newborn mice on the day of birth and on postnatal days 2, 5, and 10, under normoxic and hypoxic (5% O(2)) conditions. For both activity and apneas, visual and automatic scores showed high Pearson's correlation coefficients (0.92 and 0.98, respectively) and high intraclass correlation coefficients (0.96-0.99), supporting strong agreement between the two methods. The present results suggest that breathing disturbances may provide a valid indirect index of activity in freely moving newborn mice and that automatic apnea classification based on spectral analysis may be efficient in terms of precision and of time saved.     

新生儿抚触的研究进展

新生儿抚触作为儿科护理的新理念,是一种对新生儿健康十分有益的自然健康医疗技术。开展新生儿抚触对新生儿有特别重要的意义。抚触可以促进婴儿体格和智能的发育、增进睡眠、稳定情绪;抚触可以增强免疫力、促进食物的消化和吸收;抚触可以促进早产儿生长发育、新生儿缺氧缺血性脑病患儿的康复;抚触可以降低新生儿黄疸、避免发生胆红素脑病;抚触可促进母婴之间情感交流;抚触可利于产后妈妈的恢复。总之,无论是对家庭、社会、医院、还是对新生儿自身来讲,抚触都是非常有意义、重要且必要的,作为家长及医院工作人员都应该了解并掌握新生儿抚触的方法以及注意事项,因此,在临床工作和家庭中都应该积极主动的广泛开展新生儿抚触,目前各国都在研究并开展新生儿抚触这种护理技术,未来新生儿抚触将会被越来越多的家庭所认识并在医院中和家庭中广泛开展。     

Comparison of the Efficiency of Banna Miniature Inbred Pig Somatic Cell Nuclear Transfer among Different Donor Cells

Somatic cell nuclear transfer (SCNT) is an important method of breeding quality varieties, expanding groups, and preserving endangered species. However, the viability of SCNT embryos is poor, and the cloned rate of animal production is low in pig. This study aims to investigate the gene function and establish a disease model of Banna miniature inbred pig. SCNT with donor cells derived from fetal, newborn, and adult fibroblasts was performed, and the cloning efficiencies among the donor cells were compared. The results showed that the cleavage and blastocyst formation rates did not significantly differ between the reconstructed embryos derived from the fetal (74.3% and 27.4%) and newborn (76.4% and 21.8%) fibroblasts of the Banna miniature inbred pig (P>0.05). However, both fetal and newborn fibroblast groups showed significantly higher rates than the adult fibroblast group (61.9% and 13.0%; P<0.05). The pregnancy rates of the recipients in the fetal and newborn fibroblast groups (60% and 80%, respectively) were higher than those in the adult fibroblast group. Eight, three, and one cloned piglet were obtained from reconstructed embryos of the fetal, newborn, and adult fibroblasts, respectively. Microsatellite analyses results indicated that the genotypes of all cloning piglets were identical to their donor cells and that the genetic homozygosity of the Banna miniature inbred pig was higher than those of the recipients. Therefore, the offspring was successfully cloned using the fetal, newborn, and adult fibroblasts of Banna miniature inbred pig as donor cells.     

Electron microscopic localization of adenylate cyclase activity of white and brown adipose tissue of the rat and chicken

Summary Brown adipose tissue of newborn rats and chicken embryos and white adipose tissue of adult rats were studied. Adenylate cyclase (EC 4.6.1.1.) activity stimulated by 0.1 mmol/l noradrenaline was demonstrated using an electron microscopic histochemical method.The reaction product was visualized as a cobalt salt in the plasmalemma of the adipocytes. The adipocytes of the brown adipose tissue of the newborn rats showed most intense reaction in the outer surfaces of their plasmalemma. Alloxan totally inhibited the enzymatic reaction.The histochemical reaction used in the present study probably demonstrated the hormonal receptor sites in the plasmalemmas of the adipocytes which are stimulated by noradrenaline.     

Electron microscopic localization of adenylate cyclase activity of white and brown adipose tissue of the rat and chicken.

Brown adipose tissue of newborn rats and chicken embryos and white adipose tissue of adult rats were studied. Adenylate cyclase (EC 4.6.1.1.) activity stimulated by 0.1 mmol/l noradrenaline was demonstrated using an electron microscopic histochemical method. The reaction product was visualized as a cobalt salt in the plasmalemma of the adipocytes. The adipocytes of the brown adipose tissue of the newborn rats showed most intense reaction in the outer surfaces of their plasmalemma. Alloxan totally inhibited the enzymatic reaction. The histochemical reaction used in the present study probably demonstrated the hormonal receptor sites in the plasmalemmas of the adipocytes which are stimulated by noradrenaline.