ABSTRACT
The morphological and histological structure of the brains of Bufo gargarizans and Cynops orientalis were observed by anatomy and light microscopy. The results show that the brains of Bufo gargarizans and Cynops orientalis are divided into 5 parts which include the telencephalon, diencephalon, mesencephalon, cerebellum and medulla oblongata. The telencephalon consists of the olfactory bulb and the cerebral hemisphere. The olfactory bulb is developed that has two pairs of olfactory nerve. Bufo gargarizan has a symmetrical oval hemisphere optic lobes; Cynops orientalis only has a spherical optic lobe. The cerebellum is situated behind the optic lobe and closely connected with the myelencephalon. In this paper, the morphological and histological differences between the two species are discussed. The proportion of cerebral hemisphere is gradually increasing, which correlated with a progressive increase in the number of neuronal cell classes, and reflected in behavior complexity.
La estructura morfológica e histológica de los cerebros de Bufo gargarizans y Cynops orientalis se observó mediante anatomía y microscopía óptica. Los resultados muestran que los cerebros de Bufo gargarizans y Cynops orientalis se dividen en 5 partes, que incluyen el telencéfalo, diencéfalo, mesencéfalo, cerebelo y mielencéfalo. El telencéfalo consiste en bulbo olfatorio y hemisferio cerebral. El bulbo olfatorio tiene dos pares de nervios olfatorios. Los lóbulos ópticos de Bufo gargarizans son ovalados y simétricos en ambos hemisferios cerebrales; Cynops orientalis tiene solo un lóbulo óptico esférico. El cerebelo está situado detrás del lóbulo óptico y está estrechamente conectado con el mielencéfalo. En este trabajo, se discuten las diferencias morfológicas e histológicas entre las dos especies. El tamaño del hemisferio cerebral aumenta gradualmente, lo que se correlaciona con un aumento progresivo de células neuronales en los núcleos, reflejándose en la complejidad del comportamiento.
Subject(s)
Animals , Salamandridae/anatomy & histology , Brain/anatomy & histology , Bufo bufo/anatomy & histology , Anatomy, Comparative , Telencephalon/anatomy & histology , Mesencephalon/anatomy & histology , Cerebellum/anatomy & histology , Diencephalon/anatomy & histology , Myelencephalon/anatomy & histologyABSTRACT
As known,simultaneous determination of various chemical indicators is one of the future trends in quality control of traditional Chinese medicines because of the extremely complex chemical compositions. This project is to screen the quality markers that can accurately control the quality of the Bufonis Venenum by exploring the intrinsic correlation of components. In this study,venom of Bufo bufo gargarizans from 17 different sources were used as research samples,and the contents of 7 bufogenin were determined by HPLC-DAD. Then,the data obtained were analyzed by Spearman correlation analysis and principal component analysis( PCA). In addition,a stepwise regression analysis was used to establish a predictive model for the contents of the seven bufogenin components( independent variable) and the total contents of the bufogenin( dependent variable). The results indicated that there is a significant positive correlation between the contents of telocinobufagin and cinobufotalin,and there is a significant positive correlation between the contents of bufalin,cinobufagin and resibufogenin. In contrast,the contents of telocinobufagin and cinobufotalin are negatively correlated with the contents of bufalin,cinobufagin and resibufogenin. However,the correlation between gamabufotalin and bufotalin and other components are not obvious. Furthermore,further study found that there is a correlation between the sum of the contents of bufalin,cinobufagin and telocinobufagin and the total contents of the bufogenin. In fact,the application of bufalin,cinobufagin and telocinobufagin as the quality control indicators of the Bufonis Venenum can better reflect the quality characteristics of the Bufonis Venenum compared with the previous quality control indicators. The conclusions will provide a reference for the revision of the quality standards of the Bufonis Venenum.
Subject(s)
Animals , Amphibian Venoms , Chemistry , Bufanolides , Bufo bufo , Chromatography, High Pressure Liquid , Medicine, Chinese Traditional , Quality ControlABSTRACT
The objective of this study was to explore the relationship between the retinal structure and its life adaptation to the environment of Ctenopharyngodon idella, Cynops orientalis, Bufo bufo gargarizans, Gekko japonicus and Columba livia . Measuring retinal thickness of each layer, the nuclei layer, and the diameter of each nuclear layer of the five animals, the statistical data analysis shows that: the nuclei layers of five animals are all 4, and their structures can be divided to 10 layers when observing with optical microscope. The retinal thickness of Ctenopharyngodon idella was 190.49 mm, Cynops orientalis was 173.07 µm, and the Bufo bufo gargarizans was 195.06 µm, Gekko japonicus was 224.32 µm and Columba livia was 174.10 µm. The number of retinal inner nuclear layers of Bufo bufo gargarizans and Gekko japonicus and Columba livia are more than their outer nuclear layers, on the contrary, retinal inner nuclear layers of Ctenopharyngodon idella and Cynops orientalis are less than their outer nuclear layers. The rod and cone layer of retina of Cynops orientalis were more advanced, but their nerve fiber layer (NFL) degraded highly, revealing a strong photosensitivity but a low visual sensitivity; to Columba livia , their NFL of retina are highly developed, so as their vision. The different structures and functions of the retina of Ctenopharyngodon idella, Cynops orientalis, Bufo bufo gargarizans, Gekko japonicus and Columba livia correspond with their behavioral characteristics and the living environment's change from aquatic to amphibious to land.
El objetivo de este estudio fue explorar la relación entre las estructuras de la retina y su adaptación al medioambiente en Ctenopharyngodon idella, Cynops orientalis, Bufo bufo gargarizans,Gekko japonicus y Columba livia . La medición del espesor de cada capa de la retina, la capa nuclear y su diámetro en los cinco animales, mostró a través del análisis estadístico que las capas nucleares en todos ellos fueron 4, y sus estructuras se pueden dividir en 10 capas cuando se observan con el microscopio óptico. El espesor de la retina de Ctenopharyngodon idella fue 190,49 µm, de Cynops orientalis fue 173,07 µm, de Bufo bufo gargarizans fue 195,06 µm, de Gekko japonicus fue 224,32 µm y de Columba livia fue 174,10 µm. El número de capas nucleares internas de la retina de Bufo gargarizans, Gekko japonicus y Columba livia fue mayor que sus capas nucleares externas, mientras que las capas nucleares internas de Ctenopharyngodon idella y Cynops orientalis fueron menos que las capas nucleares externas. La capa de conos y bastones de la retina de Cynops orientalis fue más desarrollada, pero su capa de fibras nerviosas presentó una elevada degeneración, lo que muestra una gran fotosensibilidad, pero una sensibilidad visual baja. En Columba livia, la capa de fibras nerviosas de la retina estuvo muy desarrollada, y de esta manera, su visión. El grado de desarrollo de las diferentes estructuras y funciones de la retina de Ctenopharyngodon idella, Cynops orientalis, Bufo bufo gargarizans, Gekko japonicus y Columba livia está relacionada con sus características de comportamiento y el cambio de las condiciones de las vidas acuática y anfibia en la tierra.
Subject(s)
Animals , Columbidae/anatomy & histology , Retina/anatomy & histology , Salamandridae/anatomy & histology , Bufo bufo/anatomy & histology , Carps/anatomy & histology , Lizards/anatomy & histology , Adaptation to Disasters , Histology, ComparativeABSTRACT
Twelve compounds were isolated from the venom of Bufo bufo gargarizans. On the basis of their physical and chemical properties and spectral data, their structures were identified as resibufagenin (1), bufotalin (2), desacetylcinobufagin (3), 19-oxodesacetylcinobufotalin (4), cinobufotalin (5), 1beta-hydroxylbufalin (6), 12alpha-hydroxybufalin (7), bufotalinin (8), Hellebrigenin (9), telocinobufagin (10), hellebrigenol (11) and cinobufagin-3-hemisuberate methyl ester (12), respectively. Compounds 7 and 12 are new natural products.
Subject(s)
Animals , Bufanolides , Chemistry , Bufo bufo , Medicine, Chinese Traditional , Molecular Structure , Venoms , ChemistryABSTRACT
Cinobufacino injection is purified from water extraction of the skin of Bufo bufo gargarizans, which has been widely used for various cancers in clinic with significant anti-tumor effects. Bufadienolides were regarded as the main active constituents of cinobufacino injection in previous reports. In present study, 6 bufadienolides were isolated and purified from Cinobufacino injection. Their structures were identified as 3-epi-ψ-bufarenogin (1), ψ-bufarenogin (2), 3-epi-arenobufagin (3), arenobufagin (4), 3-epi-gamabufotalin (5), and 3-oxo-arenobufagin (6), separately. Among them, 1 and 3 were new compounds, 5 and 6 were new natural products. Compounds 1, 2 and compounds 3, 4 were two pairs configuration isomers at C-3, separately.
Subject(s)
Animals , Bufanolides , Chemistry , Bufo bufo , Injections , Skin , ChemistryABSTRACT
Cinobufacino injection is a significant anti-tumor medicine for the treatment of various tumors in clinic, which was made from water extraction of the skin of Bufo bufo gargarizans. In present paper, HPLC-DAD-FT-ICR-MS method was used to identify the major bufadienolides in cinobufacino for the first time. Solid-phase extraction with dichloromethane and silica was used to enrich the total bufadienolides in cinobufacino. Based on the UV and high resolution MS/MS data, 33 bufadienolides were analyzed and characterized. Among them, eight compounds were identified by comparing with standard references unambiguously. This study elucidated the major bufadienolides in cinobufacino, which provided material foundation of cinobufacino and will be benefit for the further pharmacological research.
Subject(s)
Animals , Amphibian Venoms , Chemistry , Bufanolides , Chemistry , Bufo bufo , Chromatography, High Pressure Liquid , Molecular Structure , Spectrometry, Mass, Electrospray Ionization , Tandem Mass SpectrometryABSTRACT
The present paper deals with a histological study of the blood cells of Bufo Bufo gargarizans in different months: January, March, May, July and October. The methods used are by routine blood smear in Wright stain and observation in vivo. We found that in smears and in vivo two main types of cells of the red cells: mitotic as well as amitotic. While amitotic occurs all the year round, particularly in July, mitosis so far had been seen only in July. It is also found that there are plenty of neutrophils in the blood cells of Bufo Bufo gargarizans, furthermore, the nuclei of these cells are polymorphic, especially in January and March. Meanwhile, the concentration of red cells was lowest in May and highest in January; The concentration of white blood cells was highest in October and lowest in March; As to granulocytes, eosinophils in July and October had higher proportion, while neutrophils and basophils in July; in agranulocytes, mononuclear cells reached the highest value in March, lowest in January, lymphocytes and the maximum value appeared in May, the lowest value appeared in July. Morphological changes of thrombocytes were not obvious.
Se realizó el presente estudio histológico de las células sanguíneas de Bufo Bufo gargarizans en diferentes meses del año: enero, marzo, mayo, julio y octubre. Fueron utilizados métodos de rutina por frotis de sangre con tinción de Wright y observación in vivo. Encontramos dos tipos principales de células de glóbulos rojos al frotis como también en células in vivo: mitóticas y amitóticas. Por cuanto amitosis se produce durante todo el año, sobre todo en el mes de julio, la mitosis hasta el momento se había observado solamente en julio. Además, se encontró una gran cantidad de neutrófilos en los glóbulos de Bufo Bufo gargarizans. Los núcleos de estas células son polimórficos, especialmente en enero y marzo. La concentración de glóbulos rojos era más bajo en mayo y más alta en enero; la concentración de las células blancas de la sangre fue mayor en octubre y menor en marzo. En cuanto a los granulocitos, eosinófilos estos presentaron una mayor proporción en julio y octubre, mientras que los neutrófilos y basófilos registraran una mayor proporción en el mes de julio. Los agranulocitos y las células mononucleares alcanzaron el valor más alto en marzo, y el valor más bajo en enero. Los linfocitos y el valor máximo fue registrado en mayo, el valor más bajo fue registrado en julio. No fueron evidentes los cambios morfológicos de trombocitos, lo que podría tener relación con su estabilidad.
Subject(s)
Animals , Bufo bufo/anatomy & histology , Blood Cells/ultrastructure , Time FactorsABSTRACT
Eight compounds were isolated from Bufonis periostracum by repeated column chromatography on silica gel, ODS and Sephadex LH-20 and their structures were characterized as palmitatic acid cholesteryl ester (1), cholesterol (2), 5alpha, 8alpha-epidioxycholesta-6-en-3beta-ol (3), cholest-5-en-3beta, 7beta-diol (4), cholest-7-en-3beta, 5alpha, 6beta-triol (5), 3-octaddecyloxy-1, 2-propanediol (6), isisamide (7) and bufothionine (8) on the base of spectral analysis. Compounds 1-8 were isolated from Bufonis periostracum for the first time and compounds 3, 5, 6, 7 were obtained from Bufo bufo gargarizans and Bufo genus for the first time. The bioassays showed all tested samples displayed no antitumor activity against the cell lines such as A549, BeL 7402, HGC-27 and HL-60, except the control compound bufalin.
Subject(s)
Animals , Humans , Antineoplastic Agents , Pharmacology , Bufo bufo , Metabolism , Bufonidae , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the anti-tumor and immunity activity of toad coat (Chantui), which is a new officinal part of Bufo bufo gargarizans.</p><p><b>METHOD</b>The tumor weight of S180, H22, Lewis lung cancer cell inoculated in mice were compared between the groups of mice, fed with toad coat, and those which were not (control group). The average longevity of the mice with HCA fed with toad coat was also compared with the control group. The T lymphocyte transformation and NK cell killing activity were tested and compared with the control group. The condition of the mice which were fed with great dosage of Chantui (16 g x kg(-1) x d(-1)) was observed.</p><p><b>RESULT</b>The tumor weight was remarkably reduced in the groups which were fed with toad coat compared with the control group. Tests show that toad coat can raise the activity of both T lymphocyte and NK cell. There was no obvious side-effect when the mice were fed with great dosage of toad coat.</p><p><b>CONCLUSION</b>The results show that toad coat has a strong inhibitory activity against tumors inoculated in mice and a strong enhancement of immune activity, so it could be viewed as a new valuable safe medicinal source.</p>
Subject(s)
Animals , Female , Male , Mice , Antineoplastic Agents , Bufo bufo , Cell Line, Tumor , Killer Cells, Natural , Allergy and Immunology , Mice, Inbred C57BL , Neoplasms , Drug Therapy , Allergy and Immunology , Pathology , Random Allocation , Skin , Chemistry , T-Lymphocytes , Allergy and Immunology , Tumor BurdenABSTRACT
Los plaguicidas organofosforados (OP) son masivamente aplicados en el Alto Valle de Río Negro y Neuquén, afectando al ecosistema. Utilizamos un modelo embrionario de anfibios (Rhinella arenarum) para estudiar mecanismos por los cuales OP como metilazinfos (MA) y clorpirifos (CP) podrían provocar teratogénesis. Los embriones fueron desarrollados en diferentes concentraciones de MA o CP hasta opérculo completo (OC), analizando: malformaciones, histología, glutatión reducido (GSH) y enzimas antioxidantes, poliaminas, actividad de ornitina-decarboxilasa (ODC) y proteínaquinasa-C (PKC). Ambos OP provocaron un incremento tiempo/concentración-dependiente de malformaciones, llegando a 100% de teratogénesis en estadios avanzados y a las mayores concentraciones, incluyendo: exogastrulación, curvaturas de aleta caudal, acortamiento axial, edema, y atrofia branquial. Se evidenció una condición de estrés oxidativo creciente: las enzimas GSH-dependientes (S-transferasa (GST), peroxidasa y reductasa) fueron inducidas tempranamente a bajas concentraciones, pero inhibidas en elestadio de OC a altas concentraciones, junto con una caída significativa de GSH (62%) para MA. MA incrementó significativamente (18X) la actividad de ODC en OC, aumentando los niveles de putrescina (60%) pero disminuyendo espermidina (56%) y espermina (100%); CP disminuyó en estadios tempranos la actividad de ODC y niveles de poliaminas. La disminución de poliaminas podría deberse al incremento de degradación por poliamino-oxidasa, contribuyendo al estrés oxidativo inducido por OP. Esto causaría la disminución de GSH, y la activación de PKC en OC (55%), que participaría en el control positivo de GST y ODC. Finalmente, el estrés oxidativo y la disminución en los niveles de poliaminas podrían ser causantes de alteraciones del desarrollo embrionario.
Organophosphate (OP) pesticides are widely applied in the region of Alto Valle de Río Negro y Neuquén, affecting the ecosystem. We use an amphibian embryonic model (Rhinella arenarum) in order to assess the mechanisms by which the OP pesticides azinphos methyl (AM) and chlorpyrifos (CP) could cause teratogenesis. The embryos were developed in different concentrations of AM or CP until they reached the stage of complete operculum (CO). We analyzed malformations, histology, reduced gluthatione content (GSH) and activity of antioxidant enzymes, polyamine content, ornithine decarboxilase (ODC) and protein kinase C (PKC) activities. Both OP pesticides caused a time-and dose-dependent increase in the number of malformations, reaching 100% teratogenesis in late embryonic development at the highest OP concentrations used. Malformations assessed include exogastrulation, caudalfin curvature, axial shortening, edema, and gill atrophy. Increasing evidence of oxidative stress was observed: GSH dependent enzymes (S- transferase, GST; peroxidase and reductase) were early induced in embryos exposed to low concentrations of the OP pesticides, but their activities were inhibited in the stage of CO at high concentrations of OP. These changes were accompanied by a significant decrease in GSH content (62%) in embryos exposed to AM. Besides, AM significantly increased (18X) ODC activity in the stage of CO, along with putrescine levels (60% of increase) but spermidine and spermine levels were significantly decreased (56% and 100%, respectively). The OP pesticide CP caused and early decrease in ODC activity and polyamine levels.The decrease in polyamine levels could be due to an increase in their degradation by polyamine oxidase, contributing to the oxidative stress induced by OP. This, in turn, would cause the decline in GSH levels and the activation of PKC in the embryonic stage of CO (55%), which is involved in the positive feedback of GST and ODC...
Subject(s)
Animals , Amphibians/embryology , Bufo bufo , Insecticides, Organophosphate/adverse effects , Biomarkers , Oxidative Stress , PolyaminesABSTRACT
To increase drug concentration in the head through intranasal administration, we have investigated the excised animal nasal mucosa permeability and nasal toxicity of the baicalin drug carrier systems, such as baicalin liposomes, beta-cyclodextrin inclusion compound, and phospholipid complex. A transport of baicalin drug carrier systems through nasal mucosa was simulated in diffusion chamber in vitro, and swine, caprine and rabbit nasal mucosa was used, the concentration of drug in the receptor was determined by HPLC. By taking the apparent permeability coefficients as evaluation standard, investigated the isolated animal nasal mucosa permeability of different baicalin drug systems was investigated for screening the best baicalin drug carrier system through nasal cavity administration. Toxicity of baicalin and its phospholipids complex on toad palate mucosal cilia movement and rats nasal mucosa long-term toxicity were studied in vivo. The apparent permeability coefficient of three kinds of baicalin drug carrier systems was better than that of baicalin (P < 0.05), and its lag-time was obviously shortened. At the same time, the apparent permeability coefficient of phospholipid complex was higher than those of other two drug carrier systems (P < 0.05). The results showed that the baicalin phospholipids complex nasal mucosa permeability was obviously superior to the other two drug systems. Baicalin phospholipids complex had no toxicity to ciliary movement, and had no irritation to rat nasal mucosa. The results show that baicalin phospholipid complex was the best baicalin drug carrier system, it could significantly enhance the permeability of baicalin across nasal mucosa, had no toxicity to nasal mucosa, and could be used for intranasal administration.
Subject(s)
Animals , Female , Male , Rabbits , Rats , Administration, Intranasal , Bufo bufo , Drug Carriers , Pharmacokinetics , Toxicity , Drug Delivery Systems , Flavonoids , Pharmacokinetics , Toxicity , Goats , Liposomes , Pharmacokinetics , Toxicity , Nasal Mucosa , Metabolism , Palate , Permeability , Phospholipids , Pharmacokinetics , Toxicity , Random Allocation , Swine , beta-Cyclodextrins , Pharmacokinetics , ToxicityABSTRACT
<p><b>OBJECTIVE</b>To establish a HPLC method for bufothionine in the skin of Bufo bufo gargarizans and Huachansu injection.</p><p><b>METHOD</b>The samples were separated using a Lichrosob C18 column with CH3CN-H2O (10:90) as mobile phase. Flow rate was at 1.0 mL x min (-1) and the detection wavelength was at 225 nm.</p><p><b>RESULT</b>The calibration curve of bufothionine was linear over the range of 0.0772-0.4632 microg and the average recovery was 99. 2%. The contents of bufothionine were fluctuated from 36.4-641.8 microg x g(-1) in the skin of Bufo bufo gargarizans and 22.47-33.16 microg x mL(-1) in Huachansu injection, respectively.</p><p><b>CONCLUSION</b>The contents of bufothionine were greatly different between cultured and wild species. The method was suitable for the quality control of the skin of Bufo bufo gargarizans and its preparation.</p>
Subject(s)
Animals , Bufo bufo , China , Chromatography, High Pressure Liquid , Methods , Injections , Materia Medica , Chemistry , Phenothiazines , Quality Control , Reproducibility of Results , Skin , ChemistryABSTRACT
The skin of Bufo bufo gargarizans, originated from Bufo bufo gargarizans Cantor (Bufonidae), is widely used in traditional Chinese medicine for the treatment of hepatoma, lung cancer and etc. The preparation of the aqueous components has significant therapeutic effect against the digestive tract cancer. The water-soluble chemical constituents in the skin of Bufo bufo gargarizans were then investigated to make clear the active compounds. Six compounds were isolated and purified by recrystallization and column chromatography on silica gel and ODS, their structures were elucidated as 4-amido-3-hydroxymethyl-cyclooctylamidezotetra-alpha-furanone (I), bufogargarizanine C (II), bufothionine (III), dehydrobufotenine hydrobromide (IV), suberic acid (V) and succinic acid (VI) on the basis of physicochemical properties and spectral data (UV, IR, 1H NMR, 13C NMR and MS). Of the above compounds, compounds I and II are new compounds and named bufogargarizanine B and C, respectively.
Subject(s)
Animals , Bridged Bicyclo Compounds, Heterocyclic , Chemistry , Bufo bufo , Caprylates , Chemistry , Dicarboxylic Acids , Chemistry , Medicine, Chinese Traditional , Molecular Conformation , Molecular Structure , Skin , ChemistryABSTRACT
<p><b>AIM</b>The enhancing activity and safety of several absorption enhancers were evaluated as potential nasal absorption enhancers to increase intranasal absorption of ginsenoside Rg1.</p><p><b>METHODS</b>Nasal circulatory perfusion test in vivo had been employed to investigate the effect of absorption enhancers for nasal mucosa absorption of ginsenoside Rgl in rats. The safety of the absorption enhancers were evaluated by testing cilia movement of the in situ toad palate model, the hemolysis of erythrocyte membrane of the rabbit, leaching of protein and LDH from the mice nasal mucosa and the effect on cilia structural and specific cellular changes of nasal mucosa.</p><p><b>RESULTS</b>Absorption enhancers were necessary to facilitate ginsenoside Rg1 absorption by nasal mucosa. Among the absorption enhancers 1% sodium deoxycholate had great effect to facilite ginsenoside Rgl absorption by nasal mucosa; 1% dipotassium glycyrrhizinate and 1% azone had moderate effect to facilitate ginsenoside Rg1 absorption by nasal mucosa; 1% Tween-80, 2% beta-cyclodextrin, 0.5% borneol (dissolved in paraffin liquid), 0.5% chitosan, 5% hydroxypropyl-beta-cyclodextrin and 0.1% EDTA had low effect to facilitate ginsenoside Rgl absorption by nasal mucosa. 1% sodium deoxycholate, 1% azone and 1% dipotassium glycyrrhizinate had serious nasal toxicity; 1% Tween-80, 2% beta-cyclodextrin, 5% hydroxypropyl-beta-cyclodextrin had moderate nasal toxicity; 0.5% borneol (dissolved in paraffin liquid), 0.5% chitosan and 0.1% EDTA have little nasal toxicity.</p><p><b>CONCLUSION</b>0.5% borneol and 0.5% chitosan were the promising candidates having a good balance between enhancing activity and safety for nasal ginsenoside Rg1 delivery.</p>
Subject(s)
Animals , Female , Male , Mice , Rabbits , Rats , Absorption , Administration, Intranasal , Camphanes , Pharmacology , Toxicity , Bufo bufo , Chitosan , Pharmacology , Toxicity , Cilia , Deoxycholic Acid , Pharmacology , Toxicity , Drug Synergism , Ginsenosides , Pharmacokinetics , Mice, Inbred ICR , Nasal Mucosa , Metabolism , Pathology , Rats, Sprague-DawleyABSTRACT
In this paper, membrane current properties of the fully-grown oocytes from toad, Bufo bufo gargarizans, were studied by using two-microelectrode voltage clamp technique. Axion of adult female toad was destroyed, and then ovarian lobes containing oocytes in stage I to VI were removed and incubated in Ca(2+)-free ND96 solution with collagenase (1.5 mg/ml) for 1 h. Subsequently, the oocytes were washed in Ca(2+)-free ND96 solution for 10 min to completely remove the follicular layer. For the experiments only the oocytes in stage V and VI were selected and used during 1 to 5 d. The membrane was depolarized from a holding potential of -80 mV to +60 mV in 10 mV step. It was found that a sustained outward current was elicited by depolarization. Potassium channel blockers (tetraethylammonium chloride, TEA, 10 mmol/L and 4-aminopyridine, 4-AP, 10 mmol/L) reduced the outward current to (23.4+/-0.72)% of the maximum. However, further addition of chloride channel blocker (5-nitro-2, 3-phenypropylamino benzoate, NPPB, 30 micromol/L) could almost completely block the outward current to (2.1+/-0.08)% of the maximum. In the presence of TEA and 4-AP, removal of extracellular Ca(2+) or adding verapamil (40 micromol/L), could also reduce the outward current to (2.2+/-0.04) % and (3.1+/-0.15) % of the maximum, respectively. It is concluded that calcium-dependent chloride channels exist in plasma membrane of Bufo bufo gargarizans oocytes, besides potassium channels.
Subject(s)
Animals , Female , 4-Aminopyridine , Toxicity , Bufo bufo , Calcium , Metabolism , Cell Membrane , Metabolism , Chloride Channels , Physiology , Nitrobenzoates , Pharmacology , Oocytes , Metabolism , Tetraethylammonium Compounds , Pharmacology , Verapamil , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the nasal epithelium toxicity of adjuvants and rHV2 nasal spary(HVS).</p><p><b>METHOD</b>Ciliary movement were evaluated with in situ toad palate model; The histology assessment of nasal epithelium were carried out after long-lasting and repeated use of HVS.</p><p><b>RESULT AND CONCLUSION</b>Adjuvants included SDS, Brij 35, azone, lecithin, EDTA, menthol, nipagin and thiomersal were able to significantly inhibited the ciliary movement, while tween80, glycyrrhizic acid monoammonium salt, benzalkonium bromide, sodium benzoate and adhensive materials investigated had less influence on it. HVS was able to damaged the nasal epithelium, but this effect recovered soon after stopping administration. It was demonstrated that SDS, Brij 35, azone,lecithin, EDTA, menthol, nipagin and thiomersal. It had significant cilitoxity, while tween80, glycyrrhizic acid monoammonium salt, benzalkonium bromide, sodium benzoate and adhensive materials investigated had no significance; Chitosan co-administration with some adjuvants may make the cillitoxity severer; It is available that rHV2 be administered by nasal spary.</p>
Subject(s)
Animals , Female , Male , Rabbits , Adjuvants, Pharmaceutic , Toxicity , Administration, Intranasal , Bufo bufo , Chitosan , Toxicity , Cilia , Epithelium , Hirudins , Toxicity , Nasal Mucosa , Palate , Recombinant Proteins , ToxicityABSTRACT
The classical cable function has been used to represent the response of peripheral nerves stimulated by external parallel electric field. Experiments show that peripheral nerves can be activated by perpendicular electric field induced by magnetic pulses, indicating that the activation mechanism needs to be thoroughly investigated. Several excitation properties of peripheral nerves in transverse-field were explored in the present paper. With a human's median nerve in vivo, stimulation threshold, excitation position and the relation between excitation threshold and fiber's radius were studied. The relation between stimulation threshold and stimulation duration was researched with sciatic nerves from toad in vitro. The experimental results verify that the modified cable function is much efficient than the classical cable function. The research will improve the nerve magnetic stimulating technique and be beneficial to further application.
Subject(s)
Animals , Humans , Bufo bufo , Electric Stimulation , Electromagnetic Fields , Electrophysiology , Magnetics , Median Nerve , Physiology , Peripheral Nerves , Physiology , Sciatic Nerve , PhysiologyABSTRACT
The classical cable function has been used to represent the response of peripheral nerves stimulated by external parallel electric field. It can not describe the excitation of peripheral nerves stimulated by perpendicular electric field. In this paper, responses of the nodes of Ranvier to transverse-field are deeply investigated by mathematic simulation and in-vitro experiments. The paper demonstrates that, under perpendicular electric field stimulation, the responses evoke a two-stage process including an initial polarization and the actual change of the transmembrane potential. It is the net inward current along a radial direction of the node of Ranvier that causes the peripheral nerve excitation. Based on the two-stage process, a novel model is introduced to describe peripheral nerves stimulated by transverse-field, and the classical cable function is modified. The new model and the improved cable function are verified by several in-vitro experiments. They can be used to represent peripheral nerves responses by arbitrary electric field stimulation.
Subject(s)
Animals , Bufo bufo , Electric Conductivity , Electric Stimulation , Electromagnetic Fields , Electrophysiology , Membrane Potentials , Models, Biological , Nerve Fibers , Physiology , Peripheral Nerves , Physiology , Ranvier's Nodes , Physiology , Sciatic Nerve , PhysiologyABSTRACT
The dielectric spectroscopy of the skeletal muscle cells is approached by extracellular perfusion, using curve fitting and numerical calculation with nonlinear Cole-Cole equation, and the dielectric parameter of skeletal muscle cells is compared after perfusion, The results show: (1) the permittivy at low frequency (epsilon1) and the conductivity at hight frequency (kappa(h)) reduces in keeping with extracellular perfusate; (2) the effect of extracellular perfusion on the characteristic frequency (f(c1),f(c2)) is small; (3) the extracellular perfusion mostly influences the peaks of deltaepsilon" and tgdelta for the skeletal muscle cells.
Subject(s)
Animals , Bufo bufo , Electric Conductivity , Electricity , Electrophysiology , Extracellular Fluid , Physiology , Muscle, Skeletal , Cell Biology , Physiology , Spectrum Analysis , MethodsABSTRACT
Examination of gastrointestinal contents of Bufo melanostictus collected from Bareilly, India revealed the presence of a nematode parasite Oxysomatium in 100% of the hosts sampled. The concentration index of the parasite decreased with the growth of the host [1-20 gms: 32 parasites per host, 25-50 gms.: 20 parasites per host and 50-75 gms. :2-5 parasites per host]. Its ovo-viviparous in mature, the nerve absence of ring, lateral alae and gubernaculum, the absence of preanal papillae and presence of reduced number of post anal papillae [4-6 pairs] and difference in the spicule length differentiate the present species from the earlier reported ones and is described herein as Oxysomatium bareilliana n sp