In vivo evidence of angiogenesis inhibition by ß2-glycoprotein I subfractions in the chorioallantoic membrane of chicken embryos

The vascular network expansion and functioning are important factors affecting normal intra-uterine fetal development. This study addressed the previously reported antiangiogenic potential of beta-2-glycoprotein I (β2GPI) in vivo in the chick embryo model of angiogenesis. The effects of two naturally occurring β2GPI forms on the development of the chorioallantoic membrane (CAM) vessels and the chicken embryo were investigated. β2GPI monomers and dimers were obtained by fractioned purification and characterized using SDS-PAGE, immunoblot, and ELISA. The egg exposure was performed by injection of small volumes of 2.5 µg/mL solutions of the β2GPI subfractions. Angiogenesis was evaluated through quantitative measurements of vascular architecture parameters in the captured CAM images, using computational analysis of texture contrasts and computer vision techniques. Quantitative information was assigned to the CAM vasculature modifications. In vivo, the β2GPI dimer completely halted the formation of CAM vessels and led to embryo death after 48 h of exposure. The β2GPI monomer allowed the embryo to develop up to the 10th day, despite early changes of CAM vessels. The impaired normal vessel growth proceeded as a self-limited effect. The β2GPI monomer-exposed eggs showed reduced vascularization on the 6th day of incubation, but embryos were viable on the 10th day of incubation, with ingurgitated CAM vessels implying sequelae of the angiogenesis inhibition. Both subfractions impaired CAM vasculature development. The β2GPI dimer proved to be largely more harmful than the β2GPI monomer. β2GPI modification by cleavage or dimerization may play a role in angiogenesis control in vivo.

Hey1 gene expression patterns during the development of branchial arches and facial prominences / Patrones de expresión del gen Hey1 durante el desarrollo de arcos branquiales y prominencias faciales

ABSTRACT Objective. The present study aimed to describe in detail the expression patterns of the gene Hey1, an effector of the Notch pathway, during the development of branchial arches and facial prominences. Materials and methods. Fertilized chicken (Gallus gallus) eggs obtained from a local egg farm were incubated at 37.5 -38.5ºC with 70% relative humidity until the embryos reached Hamilton-Hamburger stages HH14 through HH25. Digoxigenin-UTP labeled probes Hey1 were generated from linearized plasmids with either T3 polimerase for in vitro transcription. Whole-mount in situ hybridization was then performed. At least 3 replicates (n=3) were obtained for each stage. To confirm the results observed in whole embryos, sagittal and coronal cryosectioning was performed using a thickness of 10 µm. Results. During developmental stages HH14 and HH18, Hey1 gene expression was localized to the endoderm of branchial pouches. Hey1 gene expression was also observed in the epithelium that covers the maxillary and mandibular prominences during developmental stages HH19 and HH21, as well as in the nasal epithelium between HH19 and HH25. Transcripts were also detected in the epithelium that covers the frontonasal prominence during stage HH21. Conclusions. These expression patterns suggest the participation of this component of the Notch signaling pathway in craniofacial morphogenesis, possibly establishing pharyngeal segmentation patterns during early stages and/or regulating cell proliferation and differentiation during the late stages of facial development.

RESUMEN Objetivo. El presente estudio tuvo como objetivo describir detalladamente los patrones de expresión del gen Hey1, un efector de la vía Notch durante el desarrollo de arcos branquiales y prominencias faciales. Materiales y métodos. Se incubaron huevos fertilizados de pollo (Gallus gallus) obtenidos de una granja local entre 37.5-38.5ºC con humedad relativa del 70% hasta que los embriones alcanzaron los estadios HH14 hasta HH25 de Hamilton-Hamburger. Las sondas Hey1 marcadas con digoxigenina-UTP se generaron a partir de plásmidos linearizados con T3 polimerasa por transcripción in vitro. Luego se realizó hibridación in situ sobre embriones completos. Se obtuvieron al menos 3 repeticiones (n=3) para cada estadio. Para confirmar los resultados observados en embriones completos, se realizaron cortes sagitales y coronales de 10 µm. Resultados. Durante los estadios de desarrollo HH14 y HH18, la expresión del gen Hey1 se localizó en el endodermo de las bolsas branquiales. La expresión génica de Hey1 también se observó en el epitelio que cubre las prominencias maxilares y mandibulares durante las etapas de desarrollo HH19 y HH21, así como en el epitelio nasal entre HH19 y HH25. También se detectaron transcritos de Hey1 en el epitelio que cubre la prominencia frontonasal durante la etapa HH21. Conclusiones. Estos patrones de expresión sugieren la participación de este componente de la vía de señalización Notch en la morfogénesis craneofacial, posiblemente estableciendo patrones de segmentación faríngea durante las primeras etapas y / o regulando la proliferación y diferenciación celular durante las últimas etapas del desarrollo facial.

Construction of Wnt3a fusion protein vector and its effect on the proliferation and axon formation of neural precursor cells during the chick embryonic spinal cord development / 解剖学报

Objective To construct a eukaryotic vector of chicken-derived Wnt3a tagged with EGFP (pCAG-MCs-Wnt3a-EGFP) and investigate the influence to the proliferation and axonal formation of neural precursor cells when Wnt3a was overexpressed during the development of chick embryonic spinal cord. Methods Wnt3a gene was amplified from the total RNA obtained from chick embryonic spinal cord using molecular techniques, then connected with pCAG-MCs-EGFP to construct pCAG-MCs-Wnt3a-EGFP, which was identified by digestion and genetic sequencing. At embryonic day (E) 2.5-3.0, pCAG-MCs-Wnt3a-EGFP (experimental group) and pCAG-MCs-EGFP (control group) were transfected into the chick embryonic spinal cord using in vivo electroporation, respectively. Samples were collected at E4 (5 simples of each groups) and then conducted frozen section. The immunofluorescent staining was performed to detect the expression of Wnt3a and proliferating cell nuclear actigen (PCNA) for analyzing the relationship between Wnt3a and cell proliferation, and observe the axonal formation of neural precursor according to the green fluorescence of Wnt3a protein. Results pCAG-MCs-Wnt3a-EGFP was obtained and its gene sequencing was identical with the Gene bank. Green fluorescence was observed at E4 after pCAG-MCs-Wnt3a-EGFP transformed to chick spinal cord. In transversal section of chick embryonic spinal cord, the results of immunofluorescent staining showed Wnt3a was successfully overexpressed. Meanwhile, the amount of neurons projecting axons was dramatically decreased (n=3, P < 0.01), compared to the control group, concomitant with the significant elevation of PCNA level (n =3, P < 0.01). Conclusion pCAG-MCs-Wnt3a-EGFP is successfully constructed and our study confirmed that Wnt3a plays a vital role in the proliferation and axonal formation of neural precursor cells in the developing chick spinal cord.

Assessment of the pathogenicity of cell-culture-adapted Newcastle disease virus strain Komarov

Newcastle disease vaccines hitherto in vogue are produced from embryonated chicken eggs. Egg-adapted mesogenic vaccines possess several drawbacks such as paralysis and mortality in 2-week-old chicks and reduced egg production in the egg-laying flock. Owing to these possible drawbacks, we attempted to reduce the vaccine virulence for safe vaccination by adapting the virus in a chicken embryo fibroblast cell culture (CEFCC) system. Eighteen passages were carried out by CEFCC, and the pathogenicity was assessed on the basis of the mean death time, intracerebral pathogenicity index, and intravenous pathogenicity index, at equal passage intervals. Although the reduction in virulence demonstrated with increasing passage levels in CEFCC was encouraging, 20% of the 2-week-old birds showed paralytic symptoms with the virus vaccine from the 18^th(final) passage. Thus, a tissue-culture-adapted vaccine would demand a few more passages by CEFCC in order to achieve a complete reduction in virulence for use as a safe and effective vaccine, especially among younger chicks. Moreover, it can be safely administered even to unprimed 8-week-old birds.

Detection of chicken embryo lethal orphan virus and egg drop syndrome virus by multiplex polymerase chain reaction / 中国比较医学杂志

Objective To establish multiplex PCR assay for detection of chicken embryo lethal orphan virus (CELO)and egg drop syndrome virus (EDS).Methods According to GenBank gene sequence , two pairs of specific primers designed were amplified CELO long fiber protein and EDS hexon protein gene sequence .The specificity and sensitivity of multiplex PCR were tested .We also use the multiplex PCR to detect exogenous CELO and EDS in influenza virus.Results Two target bands have been successfully amplified and verified by sequencing .The specificity of the method is better , and the sensitivity is 10 -4μg/mL.The results of detecting exogenous CELO and EDS in 12 influenza virus were negative .Conclusion The multiplex PCR assay for detection of CELO and EDS was established successfully , which have good specificity and high sensitivity , and have high value and application prospect for detecting exogenous CELO and EDS in influenza virus .

Effects of Solanine on the human colon cancer in chicken embryo transplantation model angiogenesis / 重庆医学

Objective To establish chicken embryo transplantation model of human colon cancer and to research the effect of so‐lanine on angiogenesis .Methods Cases with chicken embryos were divided into the low‐,mid‐and high dose solanine group and con‐trol group ,with 10 cases in each groups ,and then the cultured human colon cancer cell line HT‐29 cell lines were inoculated to the chicken embryo villus allantois membrane (CAM ) .We observed the characteristics of the transplanted tumor in CAM angiogenesis by the stereo microscope .Image analysis software of Image‐pro plus 6 .0 and immunohistochemical method were used to observe the effect of different dose of solanine on angiogenesis .Results HT‐29 cell lines were inoculated to CAM 3-5 days ,a large number of blood vessels concentrated in tumors ,growing into or acrossing the surface of tumors .While tumors also rapidly growed .We took photo on the 5th day after receiving medicine and did imaging analysis .Then we calculated the area of angiogenesis in experimental group ,which was significantly lower than that of the control group ,quantitatively in a dose‐dependent manner .There were signifi‐cant differences among the groups(P<0 .01) .Microvascular density of 3 different dose of solanine was significantly lower than that of the control group by immunohistochemical method ;the expression of Ki‐67 antigen index decreased gradually ,which was highest in the control group ,and there were significant differences among the groups (P<0 .01) .Conclusion Solanine could inhibit angio‐genesis induced by human colon cancer HT‐29 cell lines obviously ,thus inhibiting the growth of tumor and providing an important basis for the treatment of anti‐tumor angiogenesis .

A new method to study nerve fiber projecting in the spinal cord during chicken embryo development / 解剖学报

Objective To develop a method of studying fiber projecting in the spinal cord duiring chicken embryo development.Methods At embryonic incubation 3 day (E3), pCAGGS-green fluorescent protein (GFP) plasmid was injected into the spinal cord using in vivo electroporation.Three days after transfection (E6), GFP-positive embryos were collected under a stereo fluorescence microscope .Subsequently , the spinal cord was separated from the embryos and cut from the roof plate as an open book .After fixed with 4%paraformaldehyde ( PFA) for one hour , the opened spinal cords were used for immunohistochemistry with N-cadherin antibody and with DAPI for nuclei .Finally, the nerve fiber projecting was photographed and analyzed under a fluorescence microscope . Results Based on the opened spinal cord and immunostaining in the cryosection , we observed that the nerve fibers projected across the midline of the floor plate and reached to the sulcus terminalis along the white matter of the contra side .The immunoreaction against N-cadherin indicated that overexpression of GFP has no significant effect on chicken embryonic development .Conclusion A new method to study fiber projecting in the developing chicken spinal cord is established successfully in this study .

The developmental toxicity of cottonseed extraction on chicken embryo

Cottonseed meal is widely used as one of the protein supplement in poultry diets. Its mechanism of toxic action on chicken embryo is poorly understood. In this study, direct effects and abnormalities of cottonseed on chicken embryo were studied. Oxidative stress, cholinergic stress, mineral analysis and microscopic lesions were analyzed in chicken embryo which injected cottonseed extraction in 0.1, 1 and 10 mg concentration (with free gossypol 0.25 ppm, 2.5 ppm and 25 ppm respectively) at day 4 of incubation. Higher group had 100 percent mortality. Serum of alive chicken embryo at day 20 of incubation were measured for FRAP (ferric reducing ability of serum), total SH groups assay, cholinesterase assay and potassium concentration. The results expressed as mean±SD show to increase oxidative stress, cholinergic stress but significant difference (p<0.05) wasn't observed between groups. The significant difference was observed in potassium concentration in serum. Some evidence of hematotoxicity such as hemorrhage and higher number of puntate reticulocytes were detected. It is concluded, hematoxicity and hyperkalemia are toxicity mechanisms that could initiate in low concentrations of cottonseed in chick embryo.

Canine Distemper Virus Utilizes Different Receptors to Infect Chicken Embryo Fibroblasts and Vero cells / 中国病毒学·英文版

Inducing animal viruses to adapt to chicken embryos or chicken embryo fibroblasts(CEF)is a common method to develop attenuated live vaccines with full security.Canine distemper virus(CDV)also does this,but the mechanisms and particular receptors remain unclear.Virus overlay protein blot assays were carried out on CEF membrane proteins,which were extracted respectively with a Mem-PERTM kit,a radioimmunoprecipitation assay buffer or a modified co-immunoprecipitation method,and revealed a common 57 kDa positive band that differed from the 42-kDa positive band in Vero cells and also from those receptors reported in lymphocytes and293 cells,indicating a receptor diversity of CDV and the possibility of the 57-kDa protein acting as a receptor that is involved in adaptive infection of CDV Kunming strain to CEF.

Reference Gene Selection for Normalization of PCR Analysis in Chicken Embryo Fibroblast Infected with H6N1 AIV / 中国病毒学·英文版

Chicken embryo fibroblasts(CEFs)are among the most commonly used cells for the study of interactions between chicken hosts and H5N1 avian influenza virus(AIV).In this study,the expression of eleven housekeeping genes typically used for the normalization of quantitative real-time PCR(QPCR)analysis in mammals were compared in CEFs infected with H5N1 AIV to determine the most reliable reference genes in this system.CEFs cultured from 10-day-old SPF chicken embryos were infected with 100 TCID50 of H5N1 AIV and harvested at 3,12,24 and 30 hours post-infection.The expression levels of the eleven reference genes in infected and uninfected CEFs were determined by real-time PCR.Based on expression stability and expression levels,our data suggest that the ribosomal protein L4(RPL4)and tyrosine 3-monooxygenase tryptophan 5-monooxygenase activation protein zeta polypeptide(YWHAZ)are the best reference genes to use in the study of host cell response to H5N1 AIV infection.However,for the study of replication levels of H5N1 AIV in CEFs,the β-actin gene(ACTB)and the ribosomal protein L4(RPL4)gene are the best references.

Carbonated water and necro-apoptotic cell death in chicken embrio low limb / Agua carbonatada y la muerte celular por necro-apoptosis en miembro inferior de embrión de pollo

Carbonated water is a fundamental part of many drinks and its effects have been studied in many pathological situations. However, cells and tissue damage as a consequence of carbonated water has not been the subject of extensive research. We assessed the short-term effects of soda on in vitro Hanging-drop culture of myoblasts and ex vivo lower limb of 8-day-old chicken embryo skeletal muscle tissue. Several groups weren designed: a) Control (Con-tyr), b) Carbonated water (Car), c) Coffe (Caf), and d) Cola beverage (Glu). The samples were observed with light microscopy and digital imaging analysis was performed. The ultra-structure of control and treated tissue were observed with electron microscopy. Immunohistochemistry techniques, such as terminal deoxynucleotidyl transferase-mediated dUTP nick- end labeling (TUNEL), TACTS Blue Label (TdT Kits) of R&D Systems were used. The myoblasts monolayers treated with soda showed plenty of eosinophilics elements. The eosinophily corresponds to higher percentage of cell death. The muscular tissue of the low limb treated with carbonated water (Car) showed calcium phosphate and collagen decreases, 53,86% and 82,95% respectively and enlarged nuclei of a higher size, with an evident loss of the parallel arrangement and fragmented nuclei. Compared to control samples, the muscular disorganization was accompanied by a positive reaction of the apoptotic bodies on TACS, also a positive reaction to ApopTacg and another positive reaction for the metalloproteases in the inter fibrillar cartilage matrix. These changes were not significant in Tyrode's solution controls, Coffee and Cola beverage groups. The morphological outcome can be apoptosis, necrosis or a mixed phenotype, suggesting that the carbonated water toxic effect might be related to these cell death processes. Further research, exploring biochemical factors will be required to elucidate necro-apoptotic cell death induced by carbonated water.

El agua carbonatada constituye una parte fundamental en muchas bebidas y su efecto ha sido estudiado en muchas situaciones patológicas. Sin embargo, el daño celular y de tejido como consecuencia del agua carbonatada no ha sido claramente investigado. El presente trabajo evalúa el efecto agudo in vitro de la soda sobre mioblastos obtenidos por cultivos en gota pendiente y el efecto sobre tejido muscular esquelético in vivo del miembro inferior de pollo de 8 días de desarrollo. Cuatro grupos de embriones fueron seleccionados al azar: a) Control (Con-tyr), b) Agua Carbonatada o Soda Club (Car), c) Café (Caf) y d) Bebida de cola (Glu). Las muestras fueron observadas por microscopía de luz. El análisis de imágenes digitales fue realizado. La ultraestructura del tejido control y tratado fue observada con Microscopía Electrónica de Transmisión (MET). La determinación de apoptosis fue realizada a través de TUNEL y l TACS Blue Label. La actividad de metaloproteasa MMP-1 fue ensayada. La población de mioblastos tratados con Soda Club mostró un elevado número de elementos eosinofílicos interpretado como un elevado número de células muertas, a diferencia del control y el grupo tratado con cafeína y bebida de Cola. En el tejido muscular se determinó una reducción de fosfatos de calcio y fibras de colágeno en una proporción de 53,86% y 82,95% respectivamente, acompañada por un desarreglo de las fibras y núcleos fragmentados, con reacción positiva para cuerpos apoptóticos y metaloproteasas en la matriz interfibrilar. Los resultados sugieren que el efecto tóxico del agua carbonatada sobre células y tejido pudiera estar vinculado con procesos combinados de muerte celular como necro-apoptosis. Se sugiere una mayor exploración de los eventos moleculares para dilucidar la combinación de los procesos de muerte celular sugerida en el presente trabajo.

Breeder age and bone development in broiler chicken embryos / Idade da matriz e desenvolvimento ósseo em embriões de frango de corte

The effect of breeder age on long bone development was studied in chicken embryos from 12 days of incubation until hatching. Fertile eggs were incubated and randomly assigned in a 2 x 6 factorial arrangement (two breeder ages - 38 and 60 weeks and six incubation days - 12, 14, 16, 18, 20, and 21). Enzymatic activity of acid and alkaline phosphatases in tibial epiphyses and weights as well as length and width in tibias and femurs of the embryos were determined. Tartrate-resistant acid and alkaline phosphatases activity in epiphyses was not affected by breeder age. Absolute weight and width of femur and tibia were larger in 60-week-old embryos compared to 38-week-old. Enzymatic activity and morphometric measurements increased with incubation day, independently of breeder age. The results showed that the process of endochondral ossification during the last two thirds of embryo development was not influenced by the age of the breeders. Although, in terms of absolute weight, the long bones of embryos from older breeders were heavier, which was associated with the larger width of the bones, but and not with their length.

O efeito da idade da matriz sobre o desenvolvimento dos ossos longos foi estudado em embriões de frango de 12 dias de incubação até o nascimento. Ovos férteis foram incubados e distribuídos em delineamento inteiramente casualizado em arranjo fatorial 2 x 6 (duas idades de matriz - 38 e 60 semanas e seis dias de incubação - 12, 14, 16, 18, 20 e 21 dias). Determinou-se a atividade enzimática das fosfatase alcalina e ácida-resistente ao tartrato no peso e nas epífises da tíbia, no comprimento e na largura da tíbia e do fêmur. A atividade das fosfatases não foi afetada pela idade da matriz. O peso absoluto e a largura de fêmur e tíbia foram maiores nos embriões das matrizes com 60 semanas de idade. Atividade enzimática e medidas morfométricas aumentaram com o dia de incubação independentemente da idade da matriz. Concluiu-se que o processo de ossificação endocondral durante os dois últimos terços de desenvolvimento embrionário não foi influenciado pela idade das matrizes. No entanto, em termos de peso absoluto, os ossos longos de embriões provenientes de matrizes velhas foram mais pesados o que foi associado à maior largura e não ao maior comprimento dos ossos.

Chicken embryo as an experimental model for the study of gastroschisis / O embrião de galinha como modelo experimental para o estudo da gastrosquise

PURPOSE: To reproduce the experimental model of gastroschisis in chicken embryos and to prove that the histopathological changes that occur in this model can be compared to those in human gastroschisis. METHODS: A total of 278 Leghorn hen (Gallus domesticus) eggs were used. The embryos were divided into three groups: the gastroschisis group, in which the umbilical cord was opened through an orifice made in the eggshell, and the intestinal loops were exposed to a mixture of amniotic liquid and allantoid; the mixture group, in which the amniotic fluid and allantoid were simply mixed without manipulating the umbilical stump and without exposing intestinal loops; and the control group which consisted of normal embryos in which no procedure was performed. The procedures were performed on the 13th day of embryo development and the study ended on the 19th day, when the intestinal loops of the embryos were removed and sent for conventional histological study and digital morphometric analysis. RESULTS: At the end of the experiment, 23 live embryos were obtained in the gastroschisis group (11.1 percent survival), and 18 of these presented exposed intestinal loops (8.7 percent success). The embryos of the gastroschisis group weighed less than those of the other two groups. The gastroschisis group also developed intestinal changes consisting of the thickening of the intestinal wall, inflammatory infiltration of the serosa and mucosa, ischemic changes in the intestinal wall and formation of a fibrin layer over the loops. These findings are characteristic of human gastroschisis and were not observed in the two other groups studied. CONCLUSION: The experimental model in chicken embryos proved able to reproduce the intestinal changes of human gastroschisis.

OBJETIVO: Reproduzir o modelo experimental da gastrosquise em embriões de galinha. MÉTODOS: Foram utilizados 278 ovos de galinha da raça Leghorn (Gallus domesticus). Os embriões foram divididos em três grupos: grupo gastrosquise, no qual, através de um orifício na casca do ovo, o cordão umbilical foi aberto e as alças intestinais expostas a uma mistura de líquidos amniótico e alantóide; grupo mistura, no qual se promovia apenas a mistura de líquidos amniótico e alantóide, sem a exposição de alças intestinais; e o grupo controle, que consistia de embriões normais e nos quais nenhum procedimento foi realizado. Os procedimentos foram feitos no 13º dia do desenvolvimento embrionário, e o estudo encerrado no 19º, quando as alças intestinais dos embriões foram removidas e encaminhadas para análise histológica convencional e análise morfométrica digital. RESULTADOS: Ao final do experimento, foram obtidos 23 embriões vivos do grupo gastrosquise (11,1 por cento de sobrevida), 18 dos quais apresentavam alças intestinais expostas (8,7 por cento de sucesso). Os embriões do grupo gastrosquise apresentaram um peso menor que os dos outros grupos. Este grupo também desenvolveu alterações intestinais consistindo em espessamento da parede, infiltrado inflamatório da serosa e mucosa, alterações isquêmicas da parede intestinal e formação de uma camada de fibrina sobre as alças. Tais achados são característicos da gastrosquise humana e não foram observados nos demais grupos. CONCLUSÃO: O modelo experimental em embriões de galinha mostrou ser capaz de reproduzir as alterações intestinais da gastrosquise humana.

Study on antiangiogenesis and antitumor of thalidomide / 中国药理学通报

Aim To study antiangiogenesis and antitumor of thalidomide.Methods In HUVECs, cell viability was determined by MTT assay;death type was observed by electron microscope; ratio of apoptosis was quantitated by flow cytometry.Angiogenesis was tested in chicken embryo chorioallantoic membrane.Effect of thalidomide on S_(180) was examined in homograft mice and microvascular counts were detected through immunochemical staining method.Results Thalidomide might inhibite the growth of HUVECs with a IC_(50) value of(22.91?1.74) ?mol?L~(-1),cells treated by thalidomide for 48 h displayed morphological characteristics of different stages associated with apoptosis,which were irregular nucleus, condensed chromatin, ballooning endoplasmic reticulum, apoptotic bodies,under electron microscope.Thalidomide might be able to cause apoptosis or necrosis of HUVECs in flow cytometry and raised positive of antiangiogenesis with increasing of dosage in chicken embryo chorioallantoic membrane. Thalidomide as a single agent might not significantly prevent tumor growth but decrease microvascular counts in tumors, however, in combination with cyclophosphamide, thalidomide could decrease dosage of cyclophosphamide and enhance antitumor of cyclophosphamide.Conclusion Thalidomide might hold back angiogenesis,as a single agent, could not significantly prevent S_(180) tumor from growing,but acted synergistically with cyclophosphamide.

INTERFERENCE OF OUABAIN IN CHICKENEMBRYO LENS GROWTH / 解剖学报

0.05).Microscopic morphology has also been observed, no visible damage could be found in the structure of lenses from eyes injected with ouabain (figs 3 and 4). Serial sections of paraffin-embedded lenses show that the number of fiber cells increased significantly in experimental samples treated with ouabain at a concentration of 0.1 ?M(table 4, P

EFFECTS OF LOW MOLECULAR WEIGHT POLYPEPTIDE EXTRACTED FROM CHICKEN EMBRYO ON LEARNING AND MEMORY IN AGING MICE INDUCED BY D-GALACTOSE / 营养学报

Objective To investigate the effects of low molecular weight polypeptide(LMP) extracted from chicken embryo on learning and memory in senile mice induced by D-galactose(Gal). Method The 16 d old chicken embryo was acid and alkali hydrolyzed,enzymolyzed and ultrafiltered to formulate LMP. 48 mice were divided randomly into four groups: control,aging model ,aging+low dose LMP,aging+high dose LMP. The aging model and two LMP groups were treated with Gal 80 mg/ (kg bw?d) by nape subcutaneous injection,while the control group with normal saline 8ml/(kg bw?d). Control and model groups were given i.g. normal saline 20 ml/(kg bw?d),and two LMP groups 10 ml or 20 ml/(kg bw?d) LMP respectively. Learning and memory of mice were tested with Morris water maze. Results The escape latency of model group was longer and the percentage of swimming distance in platform region higher than that of control group (P

THE MORPHOLOGICAL OBSERVATIONS AND BONE HISTOMOPHOMETRIC STUDY OF THE CHICKEN EMBRYO FEMUR IN EXPERIMENTAL FLUOROSIS / 解剖学报

The femur of chicken embryo in experimental fluorosis was observed under light and electron microscope, and studied by bone histomorphometric method. The results showed that most of osteoblasts were cuboid and crowded together on the surface of bone trabecula in experimental group. The osteoid was increased and mineralized bone was diminished. The bone reformation indexes were higher significantly and the bone resorption indexes did not show differences between the two groups. The cross striations of type Ⅰ collagen fibrils in osteoid layers of experimental group were indistinct and bone lamella were jumble array. Our results suggest that though fluoride has a stimulating action on the embryonic osteoblasts and enhanced the bone formation, the osteoblasts produced abnormal collagen fibrils which caused disorder of osteoid mineralization and led to osteomalacia.

MORPHOGENESIS OF THE HEART AND THE FORMATION OF THE EPICARDIUM OF CHICKEN EMBRYO / 解剖学报

The morphogenesis of the heart and the formation of the epicardium were studied with SEM, TEM and light microscopy. The results are as follows:1. There are two stages in the development of chicken heart: First is the formation of cardiac loop. In certain places, the sulcus appears outside, and the cresta appears inside during this stage. The formation of the cardiac loop indicates the establishment of the anlagen of the heart; Second is the differentiation of the anlagen, including the septation of the heart and the shift of each part of the heart. The shift of the atrium and the conus results from the shift of the right and left ventriculum. 2. The epicardium is not differcntiatcd from the myocardium, but formed from the mesenchymal cells which proliferate in front of the sinus venosus. These cells migrate and cover the entire surface of the myocardium.

THE MORPHOLOGICAL STUDY OF THE DEVELOPMENT OF ENDOCARDIAL CUSHION AND VENTRICULAR SEPTUM OF CHICKEN EMBRYO / 解剖学报

The development of the endocardial cushion and the ventricular septum of chicken heart was studied with SEM, TEM, light microscopy and histochemical methods. The results are as follows:1. The development of endocardial cushion begins from the proliferation of the endothelial cells which form cushion cells when these cells enter the cardiac jelly. Cushion cells also proceed mitosis and migrate toward the myocardium, and the endothelial cells cease proliferation when the reserve layers of cushion cells were formed. The fused endocardial cushion participates in the formation of atrioventricular orifice, the right tubercle prolongs toward the ventricular septum and eventually fuses with it.2. The cardiac jelly of endocardial cushion contains a great deal of hyaluronic acid which arranges in "trails", pioneering cushion cells migrate along the "trails" and change the component and arrangement of the matrix.3. Muscular ventricular septum is formed by aggregation of trabeculae inside the interventricular sulcus. The dorsal part of the ventricular septum fuses with the lower part of right tubercle of endocardial cushion and the ventral part fuses with the conus septum.