Responses of blood parameters and hemoglobin subtypes in plateau zokors and plateau pikas to different altitude habitats / 生理学报

The plateau zokor (Myospalax baileyi) and plateau pika (Ochotona curzoniae) are native species unique to the Qinghai-Tibetan Plateau with successful adaptation to the hypoxic environment. In this study, the number of red blood cells, hemoglobin concentration, mean hematocrit and mean volume of red blood cells were measured in plateau zokors and plateau pikas at different altitudes. Hemoglobin subtypes of two plateau animals were identified by mass spectrometry sequencing. The forward selection sites in two animals' hemoglobin subunits were analyzed by PAML4.8 program. Homologous modeling was used to analyze the effect of forward selection sites on the affinity of hemoglobin to oxygen. The adapting strategies of plateau zokors and plateau pikas to hypoxia at different altitudes were analyzed through comparing blood parameters between the two species. The results indicated that, with increasing altitudes, plateau zokors responded to hypoxia by increasing red blood cell count and decreasing red blood cell volume, while plateau pikas took the opposite strategies to plateau zokors. In erythrocytes of plateau pikas, both adult α2β2 and fetal α2ε2 hemoglobins were identified, while erythrocytes of plateau zokors only had adult α2β2 hemoglobin, however the affinities and the allosteric effects of the hemoglobin of plateau zokors were significantly higher than those of plateau pikas. Mechanistically, in the α and β subunits of hemoglobin of plateau zokors and pikas, the numbers and the sites of the positively selected amino acids as well as the side chain groups polarities and orientations of the amino acids differed significantly, which may result in the difference of the affinities to oxygen of hemoglobin between plateau zokors and pikas. In conclusion, the adaptive mechanisms to respond to hypoxia in blood properties of plateau zokors and plateau pikas are species-specific.

Comparison of the composition and content of pulmonary surfactant among plateau zokors, plateau pikas and rats / 生理学报

In the present study, the composition and content of pulmonary surfactant (PS) were analyzed to explore the hypoxia adaptation mechanism in plateau zokors (Myospalax baileyi) and plateau pikas (Ochotona curzoniae). 36 plateau zokors and plateau pikas were trapped alive at the Laji Mountain in Guide County, Qinghai Province (at the altitude of about 3 600 m), and 36 Sprague-Dawley (SD) rats were purchased from the experimental animal center of Lanzhou University (at the altitude of about 1 500 m). All animals were lavaged after laboratory anesthesia, the blood in lung tissues was fully washed out and the lung tissues were then taken out to obtain the bronchoalveolar lavage fluid by bronchoalveolar lavage. The composition and content of phospholipids in the PS of three different kinds of animals were analyzed by using high performance liquid chromatography; the protein composition, content and types in the PS were analyzed by G-250 Coomassie brilliant blue method, polyacrylamide gel electrophoresis (PAGE) and mass spectrometry; the dissolved oxygen in the PS solutions were determined by using dissolved oxygen electrode. The results showed that the total contents of phospholipids in the PS were successively increased among plateau zokors, plateau pikas and SD rats (P 0.05). The relative content of PSe had no significant differences between plateau zokors and plateau pikas (P > 0.05), but both were significantly higher than that of SD rats (P < 0.01). The serum albumin (SA) was identified in the PS of three kinds of animals, including homologous tetramer protein containing heme, which is composed of hemoglobin β subunit, in plateau zokors and plateau pikas. Immunoglobulin (Ig) heavy chain was found in PS of plateau zokors and SD rats. The content of Ig heavy chain in plateau zokor was significantly higher than that in SD rats (P < 0.01), and the content of protein containing heme was significantly higher than that in plateau pikas (P < 0.05). The amount of dissolved oxygen was successively decreased in the PS among plateau zokors, plateau pikas and SD rats (P < 0.01), but it was significantly higher than that in saline (P < 0.01). These results suggest that the total content of proteins in the PS of plateau zokors and plateau pikas was significantly higher, while the total content of phospholipids was significantly decreased. There were high content of homologous tetramer protein containing heme in the PS of plateau zokors and plateau pikas. The relative content of DPPC, the main component of phospholipids, was significantly increased in plateau zokors. The changes of PS component and content improve the adaptability of the two plateau animals in hypoxia environment.

Anatomical investigation of the measurements, shape and arterial irrigation of the adrenal gland in New Zealand rabbits / Investigação anatômica das medidas, forma e irrigação arterial da glândula adrenal em coelhos da raça Nova Zelândia

Rabbits have been used as an experimental model in many studies. These studies are important for not only for veterinary clinicians, but also for researchers in different elds. The aim of this research was to describe gross morphological measurement, shape and arterial supply of the adrenal glands in healthy New Zealand rabbits. Dissections were performed in 30 adult rabbits, 15 males and 15 females, without macroscopic adrenal pathology. Adrenal measurements were made with a digital caliper: length, width, and thickness. The origin of the adrenal arteries was also determined. Both adrenal glands were localized cranially to the respective kidneys. The mean of the right adrenal gland was 0.88 cm length, 0.42 cm width and 0.16 cm thickness; the left gland measured 0.72 cm, 0.46 cm, and 0.17 cm, respectively. The right gland was significantly more elongated than the left (p = 0.0003) and the means of the measurements did not differ between sexes. Most of the right adrenal glands had a piriform shape (73.3%), whereas most of the left gland exhibited a bean-shaped aspect (60.0%). The arterial supply was found to arise from different arteries:  lumbar, aorta, renal, caudal mesenteric, and testicular or ovarian. Comparatively, the descriptions of shape, position and arterial irrigation of the adrenal gland in rabbits are similar to those available in rodents. The data from the pres

Coelhos têm sido utilizados como modelo experimental em muitos estudos. Esses estudos são importantes não apenas para médicos veterinários, mas também para pesquisadores de diferentes campos. O objetivo desta pesquisa foi descrever as medidas morfológicas macroscópicas, forma e suprimento arterial das glândulas adrenais em coelhos saudáveis da raça Nova Zelândia. As dissecções foram realizadas em 30 coelhos adultos, 15 machos e 15 fêmeas, sem sinais macroscópicos de patologia adrenal. As medidas adrenais foram realizadas com paquímetro digital: comprimento, largura e espessura. A origem das artérias adrenais também foi determinada. Ambas as glândulas adrenais foram localizadas cranialmente nos respectivos rins. A média da glândula adrenal direita foi de 0,88 cm de comprimento, 0,42 cm de largura e 0,16 cm de espessura; a glândula esquerda mediu 0,72 cm, 0,46 cm e 0,17 cm, respectivamente. A glândula direita foi significativamente mais alongada que a esquerda (p = 0,0003) e as médias das medidas não diferiram entre os sexos. A maioria das glândulas adrenais direitas tinha uma forma piriforme (73,3%), enquanto a maioria da glândula esquerda exibia um aspecto em “forma de feijão” (60,0%). Verificou-se que o suprimento arterial provém de diferentes artérias: lombar, aorta, renal, mesentérica caudal e testicular ou ovariana. Comparativamente, as descrições de forma, posição e irrigação arterial da glândula adrenal em coelhos são semelhantes às disponíveis em roedores. Os dados da presente investigação ajudarão na interpretação de achados patológicos e / ou experimentais em coelhos da raça Nova Zelândia.

Prevalencia y hallazgos radiográficos en conejos, cuyos y chinchillas diagnosticados con maloclusión en el Hospital Veterinario de Especialidades en Fauna Silvestre y Etología Clínica de la UNAM, México / Prevalence and radiographic findings in rabbits, guinea pigs and chinchillas diagnosed with malocclusion at the UNAM Exotic Pet and Wildlife Teaching Hospital, Mexico

RESUMEN Los lagomorfos y los histricomorfos, como los cuyos y las chinchillas, comparten características dentales como: dentadura elodonta e hipsodonta. Estas peculiaridades predisponen la presentación de maloclusión debido al consumo de dietas inadecuadas, poco abrasivas. El propósito de este trabajo es identificar la prevalencia de maloclusión en cada una de las especies mencionadas, de acuerdo con su sexo y grupo etario. Además, tiene como objetivo hacer una evaluación objetiva de los estudios radiográficos de los ejemplares, basada en líneas de referencia anatómica, para determinar los principales hallazgos radiográficos según el grado de maloclusión diagnosticado de forma clínica. El 30% de todos los individuos evaluados presentaron problemas de maloclusión, 42,86% de ellos conejos, 16,6% cuyos y 23% chinchillas. No se evidenciaron diferencias significativas en la presentación por sexo. En conejos fue más común el diagnóstico de ejemplares del grupo etario I y II, mientras que en cuyos lo fue en los grupos II y III. El grado I y II fueron los más diagnosticados en conejos y cuyos. En las radiografías las lesiones más comunes fueron el sobrecrecimiento de las raíces maxilares y mandibulares, elongación de incisivos, pérdida de la superficie de oclusión y deformación de las raíces mandibulares. Se propone un método de corrección para las proyecciones radiográficas LL con rotación evidente para mantener la objetividad de la evaluación radiográfica.

ABSTRACT Lagomorphs and histricomorphs, as guinea pigs and chinchillas, have common dental characteristics, such as the elodont and hypsodont teeth. These peculiarities predispose to the presentation of malocclusion due to the consumption of inadequate, non-abrasive diets. The purpose of this work is to identify the prevalence of malocclusion in each case. In addition, it aims to make an objective evaluation of the radiological studies of the specimens, based on anatomical reference lines, to determine the main radiographic findings according to the degree of malocclusion clinically diagnosed. 30% of the individuals evaluated presented malocclusion problems being 42.86% rabbits, 16.6% guinea pigs and 23% chinchillas. There is no evidence of significant differences in the presentation by sex. In rabbits, the diagnosis of the specimens from the group I and II were more common, while in guinea pigs they were II and Ill. Grade I and II were the most diagnosed in rabbits and guinea pigs. In the radiographs, the most common lesions are the overgrowth of the maxillary and mandibular roots, elongation of the incisors, loss of the occlusal surface and deformation of the mandibular roots. A method of correction for LL radiographic projections with evident rotation is proposed to maintain the objectivity of the radiographic evaluation.

Selected ophthalmic and physiologic parameters in rabbits (Oryctolagus cuniculus) submitted to retrobulbar block with lidocaine, morphine or ketamine / Avaliação de parâmetros oftálmicos e fisiológicos em coelhos (Oryctolagus cuniculus) submetidos a bloqueio retrobulbar com lidocaína, morfina ou cetamina

The aim of the present study was to evaluate selected ophthalmic and physiologic parameters in rabbits submitted to retrobulbar blockade with lidocaine, morphine or ketamine. Eighteen adult rabbits, seven males and eleven females, New Zealand White breed, weighing 3.9 ± 0.7 kg were randomly assigned to perform the retrobulbar block according to the groups: LID (2% lidocaine without a vasoconstrictor - 7 mg kg-1); MOR (1% morphine - 1 mg kg-1) or KET (10% Ketamine - 5 mg kg-1). Ophthalmic and physiologic parameters were assessed, including lacrimal production using Schirmer tear test (STT), corneal touch threshold (CTT), pupillary diameter, intraocular pressure (IOP), pulse rate (PR), respiratory rate (RR), oxyhemoglobin saturation (SpO2), rectal temperature (RT) and systolic, mean and diastolic blood pressures (SAP, MAP and DAP) and were evaluated every 10 minutes for 70 minutes. All drugs used in the present study promoted central positioning of the eyeball for up to one minute later the retrobulbar administration in all cases. There was a significant increase of STT values in MOR and LID, when compared to baseline, while the CTT values had a significant decrease in all groups. KET kept the IOP values unaltered at the time points and there was a significant decrease of pupillary diameter in MOR. There was no significant change in PR, RR and SpO2; however, LID presented significantly lower values of SAP. MOR had increased values for RT when compared to the other two groups. The established parameters may help in ophthalmic procedures using retrobulbar nerve blocks.

Objetivou-se com o presente estudo estabelecer parâmetros oftálmicos e hemodinâmicos em coelhos submetidos ao bloqueio retrobulbar com lidocaína, morfina ou cetamina. Dezoito coelhos da raça Nova Zelândia, adultos, sete machos e onze fêmeas, com peso de 3,9 ± 0,7 kg foram aleatoriamente distribuídos para realização de bloqueio retrobulbar de acordo com os seguintes grupos: LID (lidocaína 2% sem vasoconstrictor ­ 7 mg/kg); MOR (morfina 1% - 1 mg/kg) ou KET (cetamina 10% ­ 5 mg/kg). Os seguintes parâmetros oftálmicos e hemodinâmicos foram avaliados: produção lacrimal através do teste lacrimal de Schirmer (TLS), limiar de sensibilidade corneana ao toque, diâmetro pupilar, pressão intraocular (PIO), frequência de pulso (FP) e respiratória (FR), saturação da oxihemoglobina (SpO2), temperatura retal (TR) e pressão arterial sistólica, diastólica e média (PAS, PAD e PAM). Todos os fármacos utilizados no presente estudo promoveram centralização do bulbo do olho em até um minuto após a sua administração retrobulbar em todos os casos. Houve um aumento significativo do TLS no grupo MOR e LID, quando comparados aos valores basais, enquanto o limiar de sensibilidade corneana reduziu significativamente em todos os grupos. O grupo KET manteve os valores da PIO inalterados em todos os tempos e houve uma redução significativa do diâmetro pupilar no grupo MOR. Não houve alteração significativa dos valores de FP, FR e SpO2. No entanto, o grupo LID apresentou valores significativamente menores de PAS. O grupo MOR apresentou maiores valores de TR quando comparado aos demais grupos. Os parâmetros estabelecidos no presente estudo podem servir como base para a realização de procedimentos oftálmicos que requerem o uso de bloqueio retrobulbar.

Método imunomagnético associado ao meio MesenCult® na obtenção de células mononucleares da medula óssea de coelhos negativas para o anticorpo monoclonal CD45 / Immunomagnetic method associated with MesenCult® medium to obtain bone marrow mononuclear cells from rabbits negative for CD45 monoclonal antibody

O objetivo detse artigo é de descrever um protocolo de isolamento das células mononucleares da medula óssea de coelhos, seguido de purificação celular por depleção negativa com o anticorpo monoclonal CD45 e posterior expansão em meio de cultura MesenCult®. Dez coelhos machos adultos, da raça Nova Zelândia, com idade média de 1,0±0,2 anos e peso médio 3,5±0,24kg, foram utilizados para padronização da metodologia. O isolamento das células mononuclares da medula óssea foi realizado pelo gradiente de densidade Ficoll-paque® e a purificação e obtenção das células- pela depleção negativa com o anticorpo monoclonal CD45 em base imunomagnética. A população celular obtida foi expandida posteriormente em meio de cultura MesenCult®. No isolamento pelo gradiente de icoll-Paque® foi obtido um rendimento médio de 7,31x106 células/mL. Após purificação e obtenção das possíveis células-tronco mesenquimais pela base imunomagnética, houve um decréscimo do rendimento para 2,28x106 células/mL, mas o processo de expansão foi incrementado pelo cultivo celular. Os resultados indicaram que as células obtidas da fração mononuclear da medula óssea, cultivadas in vitro foram capazes de gerar células aderentes 24 horas após o cultivo, com predominância de células fibroblastóides sugestivas de células-tronco mesenquimais. Concluiu-se que a obtenção de células-tronco mesenquimais pode ser alcançada após purificação das células mononucleares da medula óssea de coelhos pelo método imunomagético, o meio de cultura MesenCult® proporciona um ambiente adequado para a rápida expansão in vitro e o número de passagens exerce influência negativa sobre as características morfológicas das células.

The objective of this study was to describe guidelines for the isolation of bone marrow mononuclear cells from rabbits, followed by cell purification by negative depletion with CD45 monoclonal antibody, and further expansion in MesenCult® medium. Ten adult male New Zealand White rabbits, age average of 1.0±0.2 years and weighting 3.5±0.24kg, were used to obtain a standardized method. The mononuclear cells of the bone marrow were isolated with Ficoll-paque® density gradient centrifugation, and the cell purification and acquisition was completed by negative depletion with CD45 monoclonal antibody in immunomagnetic base. The cell population obtained was expanded in MesenCult® medium. Through isolation with Ficoll-paque® density gradient was possible to obtain an average yield of 7.31x106 cells/mL. After purification and acquisiton of potential mesenchymal stem cells by the immunomagnetic base, there was a yield decrease to 2.28x106 cells/mL; however the expansion process was increased in cell culture. The results indicated that cells obtained from the mononuclear fraction of bone marrow and cultivated in vitro were capable to generate adherent cells 24 hours after culture, with predominance of fibroblastoid cells suggestive of mesenchymal stem cells. It can be concluded that mesenchymal stem cells can be achieved with purified rabbit bone marrow mononuclear cells through the immunomagnetic method, as the MesenCult® medium provides a suitable environment for a quick in vitro expansion, and the number of passages exerts negative influence on the morphological characteristics.

The expression of the sperm-specific lactate dehydrogenase gene Ldh-c in plateau pika (Ochotona curzoniae) cardiac muscle and its effect on the anaerobic glycolysis / 生理学报

The plateau pika (Ochotona curzoniae) has a strong adaptability to hypoxic plateau environment. We found that the sperm-specific lactate dehydrogenase (LDH-C4) gene Ldh-c expressed in plateau pika cardiac muscle. In order to shed light on the effect of LDH-C4 on the anaerobic glycolysis in plateau pika cardiac muscle, 20 pikas were randomly divided into the inhibitor group and the control group, and the sample size of each group was 10. The pikas of inhibitor group were injected with 1 mL 1 mol/L N-isopropyl oxamate, a specific LDH-C4 inhibitor, in biceps femoris muscle of hind legs, each leg with 500 μL. The pikas of control group were injected with the same volume of normal saline (0.9% NaCl). The mRNA and protein expression levels of Ldh-c gene in plateau pika cardiac muscle were determined by real-time PCR and Western blot. The activities of LDH, and the contents of lactate (LD) and ATP in cardiac muscle were compared between the inhibitor group and the control group. The results showed that 1) the expression levels of Ldh-c mRNA and protein were 0.47 ± 0.06 and 0.68 ± 0.08, respectively; 2) 30 min after injection of 1 mL 1 mol/L N-isopropyl oxamate in biceps femoris muscle, the concentration of N-isopropyl oxamate in blood was 0.08 mmol/L; 3) in cardiac muscle of the inhibitor group and the control group, the LDH activities were (6.18 ± 0.48) U/mg and (9.08 ± 0.58) U/mg, the contents of LD were (0.21 ± 0.03) mmol/g and (0.26 ± 0.04) mmol/g, and the contents of ATP were (4.40 ± 0.69) nmol/mg and (6.18 ± 0.73) nmol/mg (P < 0.01); 5) the inhibition rates of N-isopropyl oxamate to LDH, LD and ATP were 31.98%, 20.90% and 28.70%, respectively. The results suggest that Ldh-c expresses in cardiac muscle of plateau pika, and the pika cardiac muscle may get at least 28% ATP for its activities by LDH-C4 catalyzed anaerobic glycolysis, which reduces the dependence on oxygen and enhances the adaptation to the hypoxic environments.

Establishment of RP-HPLC detection method of N-isopropyl oxamate in the serum of plateau pikas / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the intergrating of N-isopropyl oxamate and serum protein and establish a high performance liquid chromatography (HPLC) detection method of N-isopropyl oxamate (specific inhibitor of testis-specific lactate dehydrogenase (LDH-C4)) in the blood of plateau pikas.</p><p><b>METHODS</b>Twenty highland pika 150-200 g, were randomly divided into two groups (n = 10): control group and inhibitor group. Different concentrations of N-isopropyl oxamate were added to examine the intergrating of N-isopropyl oxamate and serum protein. In order to determine its concentration in the pika blood accurately, we used the method of adding trypsin to incubate the serum first, followed by trichloroacetic acid treatment and detecting by HPLC. Results: When the concentrations of N-isopropyl oxamate in the pika serum were added to 0.05 mmol/L, 0.1 mmol/L, 1 mmol/L, 10 mmol/L, 16.7 mmol/L, 33.3 mmol/L and 100 mmol/L, the intergrating rates between N-isopropyl oxamate and plateau pika serum were 100%, 100%, 100%, 86.84%, 54.11%, 40.10% and 20.18%, respectively. The method established in this paper was good on recovery rates, precision and stability. A good linearity was obtained in the range of 0.0125-0.25 mmol/L. When the concentrations of N-isopropyl oxamate in the serum were added to 0. 15 mmol/L,0.3 mmol/L and 1 mmol/L, the recovery rates were 98.05%, 98.98% and 98.12%, respectively; the precision relative standard deviation( RSD) of concentrations were 1.17%, 0.92% and 0.83%, respectively; the stability relative standard deviation (RSD) of concentrations were 1.38%, 1.40% and 0.88%, respectively. The repeatability RSD of the method was 1.76%. Quantitative limit was 0.0125 mmol/L.</p><p><b>CONCLUSION</b>N-isopropyl oxamate has a strong affinity with plateau pika serum protein that can't be accurately determined with common HIPLC method. It can be accurately determined in the blood by adding trypsinto digest the serum protein first, followed by adding trichloroacetic acid to precipitate the protein.</p>

Bartonella Species Detected in the Plateau Pikas (Ochotona curzoiae) from Qinghai Plateau in China / 生物医学与环境科学（英文）

Bartonella species can infect a variety of mammalian hosts and cause a broad spectrum of diseases in humans, but there have been no reports of Bartonella infection in Ochotonidae. This is the first study to detect Bartonella in plateau pikas in the Qinghai plateau, providing baseline data for the risk assessment of human Bartonella infection in this area. We obtained 15 Bartonella strains from 79 pikas in Binggou and Maixiu areas of Qinghai with a positive rate of 18.99%. Based on the phylogenetic analysis of the Bartonella citrate synthase (gltA) gene sequences, most strains were closely related to B. taylorii (3/15) and B. grahamii (12/15). The latter is a pathogenic strain in humans. Our results suggest that a corresponding prevention and control strategy should be taken into consideration in the Qinghai province.

The plateau zokors' learning and memory ability is related to the high expression levels of foxP2 in the brain / 生理学报

Plateau zokor (Myospalax baileyi) is a subterranean mammal. Plateau zokor has high learning and memory ability, and can determine the location of blocking obstacles in their tunnels. Forkhead box p2 (FOXP2) is a transcription factor implicated in the neural control of orofacial coordination and sensory-motor integration, particularly with respect to learning, memory and vocalization. To explore the association of foxP2 with the high learning and memory ability of plateau zokor, the cDNA of foxP2 of plateau zokor was sequenced; by using plateau pika as control, the expression levels of foxP2 mRNA and FOXP2 protein in brain of plateau zokor were determined by real-time PCR and Western blot, respectively; and the location of FOXP2 protein in the brain of plateau zokor was determined by immunohistochemistry. The result showed that the cDNA sequence of plateau zokor foxP2 was similar to that of other mammals and the amino acid sequences showed a relatively high degree of conservation, with the exception of two particular amino acid substitutions [a Gln (Q)-to-His (H) change at position 231 and a Ser (S)-to-Ile (I) change at position 235]. Higher expression levels of foxP2 mRNA (3-fold higher) and FOXP2 protein (>2-fold higher) were detected in plateau zokor brain relative to plateau pika brain. In plateau zokor brain, FOXP2 protein was highly expressed in the cerebral cortex, thalamus and the striatum (a basal ganglia brain region). The results suggest that the high learning and memory ability of plateau zokor is related to the high expression levels of foxP2 in the brain.

Contraction responses of isolated aortic rings of pika (Ochotona curzoniae) and Sprague-Dawley rat to hypoxia / 生理学报

The aim of the present study was to observe the effects of hypoxia on tensions of aortic rings of pika (Ochotona curzoniae) and Sprague-Dawley (SD) rat. The aortic rings were prepared, and in vitro vascular ring perfusion was used to assay the effects of hypoxia or different drugs on contraction responses of the rings with or without endothelium. The results showed that, there was no difference of the contractions to KCl (80 mmol/L) between the aortic rings of the pikas and SD rats. After pre-contraction with NE (1 μmol/L), the aortic rings with endothelium of the SD rats showed obvious relaxation to ACh (1 μmol/L), whereas the aortic rings of the pikas, no matter with or without endothelium, showed significant and unusual contraction to ACh. The aortic rings of pikas, no matter with or without endothelium, exhibited greater contraction when treated by 1 h of hypoxia, compared with those in SD rats; The similar result was showed under hypoxia in combination with Ca(2+) removal. These results suggest that the contraction response to hypoxia in pika is more sensitive compared to that in SD rat, which is dependent on the release of calcium from intracellular calcium store.

Differences of glycolysis in skeletal muscle and lactate metabolism in liver between plateau zokor (Myospalax baileyi) and plateau pika (Ochotona curzoniae) / 生理学报

The plateau pika (Ochotona curzoniae) and plateau zokor (Myospalax baileyi) are specialized native species of the Qinghai-Tibetan plateau. The goal of this study was to examine physiological differences in skeletal muscle glycolysis and hepatic lactate metabolism between these two species. The partial sequence of pyruvate carboxylase (PC) gene was cloned and sequenced. The mRNA expression levels of PC and lactate dehydrogenases (LDH-A, LDH-B) were determined by real-time PCR. The enzymatic activity of PC was measured using malic acid coupling method. The concentration of lactic acid (LD) and the specific activities of LDH in liver and skeletal muscle of two species were measured. The different isoenzymes of LDH were determined by native polyacrylamide gel electrophoresis (PAGE). The results showed that, (1) LDH-B mRNA level in skeletal muscle of plateau zokor was significantly higher than that of plateau pika (P < 0.01), but no differences was found at LDH-A mRNA levels between them (P > 0.05); (2) PC, LDH-A and LDH-B mRNA levels in liver of plateau pika were significantly higher than those of plateau zokor (P < 0.01); (3) The LDH activity and concentration of LD in skeletal muscle and liver, as well as the PC activity in liver of plateau pika were significantly higher than those of plateau zokor (P < 0.01); (4) The isoenzymatic spectrum of lactate dehydrogenase showed that the main LDH isoenzymes were LDH-A4, LDH-A3B and LDH-A2B2 in skeletal muscle of plateau pika, while the main LDH isoenzymes were LDH-AB3 and LDH-B4 in skeletal muscle of plateau zokor; the main isoenzymes were LDH-A3B, LDH-A2B2, LDH-AB3 and LDH-B4 in liver of plateau pika, while LDH-A4 was the only isoenzyme in liver of plateau zokor. These results indicate that the plateau pika gets most of its energy for sprint running through enhancing anaerobic glycolysis, producing more lactate in their skeletal muscle, and converting lactate into glucose and glycogen in the liver by enhancing gluconeogenesis. As a result, the plateau pika has a reduced dependence on oxygen in its hypoxic environment. In contrast, plateau zokor derives most of its energy used for digging activity by enhancing aerobic oxidation in their skeletal muscle, although they inhabit hypoxic underground burrows.

Functional difference of malate-aspartate shuttle system in liver between plateau zokor (Myospalax baileyi) and plateau pika (Ochotona curzoniae) / 生理学报

To explore the adaptive mechanisms of plateau zokor (Myospalax baileyi) to the enduring digging activity in the hypoxic environment and of plateau pika (Ochotona curzoniae) to the sprint running activity, the functional differences of malate-aspartate shuttle system (MA) in liver of plateau zokor and plateau pika were studied. The ratio of liver weight to body weight, the parameters of mitochondria in hepatocyte and the contents of lactic acid in serum were measured; the open reading frame of cytoplasmic malate dehydrogenase (MDH1), mitochondrial malate dehydrogenase (MDH2), and the partial sequence of aspartate glutamate carrier (AGC) and oxoglutarate malate carrier (OMC) genes were cloned and sequenced; MDH1, MDH2, AGC and OMC mRNA levels were determined by real-time PCR; the specific activities of MDH1 and MDH2 in liver of plateau zokor and plateau pika were measured using enzymatic methods. The results showed that, (1) the ratio of liver weight to body weight, the number and the specific surface of mitochondria in hepatocyte of plateau zokor were markedly higher than those of plateau pika (P < 0.01 or P < 0.05), but the content of lactic acid in serum of plateau pika was significantly higher than that of plateau zokor (P < 0.01); (2) MDH1 and MDH2 mRNA levels as well as their enzymatic activities in liver of plateau zokor were significantly higher than those of plateau pika (P < 0.01 or 0.05), AGC mRNA level of the zokor was significantly higher than that of the pika (P < 0.01), while no difference was found at OMC mRNA level between them (P > 0.05); (3) mRNA level and enzymatic activity of MDH1 was significantly lower than those of MDH2 in the pika liver (P < 0.01), MDH1 mRNA level of plateau zokor was markedly higher than that of MDH2 (P < 0.01), but the activities had no difference between MDH1 and MDH2 in liver of the zokor (P > 0.05). These results indicate that the plateau zokor obtains ATP in the enduring digging activity by enhancing the function of MA, while plateau pika gets glycogen for their sprint running activity by increasing the process of gluconeogenesis. As a result, plateau pika converts the lactic acid quickly produced in their skeletal muscle by anaerobic glycolysis and reduces dependence on the oxygen.

Distribution of the intermediate hosts of Echinococcus multilocularis in Shiqu County, Sichuan, China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The rodentia and lagomorpha animals are the intermediate hosts of Echinococcus multilocularis, their distribution and infection of this parasite may facilitate the infection of definitive hosts such as dogs. The aim of this study was to determine the distribution of the intermediate hosts of Echinococcus multilocularis in Shiqu County, Sichuan, China.</p><p><b>METHODS</b>A systematic sampling method was used to investigate the density of burrows of rodents and lagomorphs at 97 pasture sites in winter and summer pastureland and remote sensing (RS) technology was used to correlate their densities to the distribution of these animals in different landscape types.</p><p><b>RESULTS</b>Based on the densities of Ochotona curzoniae, Microtus fuscus (dependent variable) and their burrow densities (independent variable) in survey points, regression equations were fitted respectively (Ochotona curzoniae, P < 0.0001, R(2) = 0.8705; Microtus fuscus, P < 0.0001, R(2) = 0.9736). Their burrow density in summer pastureland was higher than in winter pastureland (F = 36.65, P < 0.0001). The burrow densities of Ochotona curzoniae and Microtus fuscus in bareland and half-bareland are higher than in grassland (F = 7.73, P < 0.001).</p><p><b>CONCLUSIONS</b>The regression relationship between the densities of Ochotona curzoniae and Microtus fuscus and their burrow densities indicate that the burrow densities could reflect the animal densities and that the burrow density was greater in summer pastureland than in winter pastureland. The main distribution areas of the intermediate hosts were in bareland and half-bareland.</p>

Avaliação da densidade óssea e da resistência à fratura entre enxertos alógenos com fosfato de cálcio bifásico e tecido ósseo em tíbias de coelhos / Evaluation of bone density and fracture resistance between allograft with biphasic calcium phosphate and bone tissue in rabbit tibiae

Objetivo: avaliar por meio da tomografia computadorizada de feixe cônico (TCFC) a densidade do tecido ósseo formado ao redor de implantes curtos e a resistência à fratura por meio da máquina de ensaio universal de força, quando estes estão associados ou não aos enxertos de fosfato de cálcio bifásico. Método: para este estudo, foram utilizadas 10 tíbias de coelhos da raça Nova Zelândia. No grupo controle (GC) foram instalados dois implantes cilíndricos, com interface cone morse 4.0 x 6.0 mm na tíbia direita e no grupo teste (GT) foram instalados dois implantes cilíndricos, com interface cone morse 4.0 x 6.0 mm e enxerto com o biomaterial fosfato de cálcio bifásico na tíbia esquerda. Após o sacrifício dos animais e remoção das tíbias, estas foram encaminhadas para a realização do exame de TCFC e teste de resistência à fratura. Resultados: após análise dos dados verificaram-se que as médias em escala Houns para os espécimes com enxerto foram pontualmente maiores que para os espécimes sem enxerto (948,55 e 739,06 respectivamente) e comprovaram-se diferenças estatisticamente significantes entre os espécimes. Com relação ao teste de resistência, as médias em Newton para os GC e GT foram 15,586 e 16,028 respectivamente. Conclusão: o GT apresentou as maiores densidades ósseas e esse aumento foi verificado principalmente nos dois primeiros milímetros, local onde efetivamente ocorreu a colocação do enxerto. Com relação à resistência à fratura, os grupos controle e teste apresentaram resultados similares.

Objective: to evaluate by cone beam computed tomography (CBCT) the density of bone tissue around implants short and resistance to fracture by a universal testing machine of force when they are associated or not to graft phosphate biphasic calcium. Method: for this study, we used 10 rabbits tibias of New Zealand in the control group (CG) were made in two cylindrical implants, Morse taper interface with 4.0 x 6.0 mm in the right tibia and in the test group (TG) were made in two cylindrical implants, Morse taper interface with 4.0 x 6.0 mm and graft with the biphasic calcium phosphate biomaterial in the left tibia. After sacrificing the animals and removal of tibia, these were referred to the examination of CBCT and fracture resistence test. Results: data analysis showed that the average scale Houns for specimens with grafts were briefly higher than for specimens without graft (948.55 and 739.06 respectively), proved to be statistically significant differences between the specimens. With respect to the test of resistance to the averages in Newton for the CG and TG were 15.586 and 16.028 respectively. Conclusion: the TG had the highest bone density and this increase was mainly observed in the first two millimeters, where effectively there placing the graft. With respect to resistance to fracture, the test and control groups showed similar results.

Biological parameters of larvae, nymphs, and engorged females of Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae) obtained from artificial and successive infestations on Oryctolagus cuniculus (Linnaeus, 1758) (Lagomorpha: Ochotonidae) / Parâmetros biológicos de larvas, ninfas e fêmeas ingurgitadas de Amblyomma cajennense (Fabricius, 1787) (Acari:Ixodidae) obtidas de infestações artificiais e sucessivas em Oryctolagus cuniculus (Lagomorpha:Ochotonidae)

The development of acquired resistance to Amblyomma cajennense ticks was evaluated. A total of 18 rabbits were uniformly distributed into three groups named GL, GN, and GA that were submitted to three consecutive infestations with, respectively, larvae, nymphs, and adults of A. cajennense. After the infestations, the parasitic and reproductive parameters of recovered instars were evaluated. A significant reduction of recovery rates of inoculated ticks was observed only after the third infestation of animals in groups GL and GN (P<0.01). The modal day for drop-off of engorged larvae and nymphs increased along the infestations. The recovered engorged nymphs presented a significant reduction of the mean body weight between the first and the third infestations (P<0.01). A significant reduction in the mean recovery rate of engorged females was only observed during the third infestation. However, significant reductions (P<0.05) in body weight, egg mass weight, and larva eclosions were observed for recovered engorged females after the second infestation onwards. The results suggest the occurrence of resistance of rabbits to A. cajennense infestations, being more evident against the adult stage.

Avaliou-se o desenvolvimento de resistência adquirida frente ao parasitismo por Amblyomma cajennense. Para tanto, 18 coelhos foram uniformemente distribuídos em três grupos denominados GL, GN e GA e submetidos a três infestações consecutivas por, respectivamente, larvas, ninfas ou adultos de A. cajennense. Após a infestação, estudaram-se os parâmetros parasitários e reprodutivos dos exemplares recuperados. Apenas a partir da terceira infestação dos animais dos grupos GL e GN, pôde-se observar uma queda significativa na taxa de recuperação dos ixodídeos inoculados (P<0,01). O dia modal de queda das larvas e ninfas recuperadas aumentou com o decorrer das infestações. As ninfas ingurgitadas recuperadas apresentaram uma redução significativa (P<0,01) no peso médio corporal entre a primeira e terceira infestação. Observou-se uma queda significativa na taxa média de recuperação das fêmeas ingurgitadas somente na terceira infestação (P<0,01). Todavia, a partir da segunda infestação observou-se nas teleóginas recuperadas, uma redução significativa no peso corporal, peso da postura e eclodibilidade das larvas (P<0,05). Os dados sugerem a ocorrência de resistência em coelhos à infestação por A. cajennense que foi mais evidente contra o estádio adulto.

Difference in oxygen uptake in skeletal muscles between plateau zokor (Myospalax rufescens baileyi) and plateau pika (Ochotona curzoniac) / 生理学报

To investigate the difference between the functions of oxygen uptake in skeletal muscle and living habits of plateau zokor (Myospalax rufescens baileyi) and plateau pika (Ochotona curzoniac), the microvessel densities (MVD) of skeletal muscle of plateau zokor, plateau pika and Sprague-Dawley (SD) rat were measured by immunohistochemical staining; the numerical density on area (N(A)) of mitochondria, and surface density (S(V), external surface area density of mitochondria per unit volume of skeletal muscle fiber) were obtained by stereo microscope technique; mRNA levels of myoglobin (Mb) in skeletal muscle were determined by real-time PCR, and the contents of Mb protein in skeletal muscle were determined by spectro-photometer. The results showed that MVD, N(A) and S(V) of mitochondria in skeletal muscle of plateau pika were significantly lower than those of plateau zokor and SD rat (P<0.05). The mRNA levels of Mb gene in skeletal muscle of plateau zokor and plateau pika were notably higher than that of SD rat (P<0.05). There were significant differences in the contents of Mb among these three species, and plateau zokor and SD rat presented the highest and the lowest value, respectively (P<0.05). The results suggest that even though plateau zokor inhabits in the hypoxia environment, most of its skeletal muscle fiber are red muscle fiber. While most of skeletal muscle fibers of plateau pika are white muscle fibers. This kind of white muscle has low MVD, N(A) and S(V) of mitochondria and less content of Mb compared with the red one, suggesting it obtains most energy from aerobic oxidation. The above-mentioned differences in skeletal muscles may be related to not only the different species, but also the different living habits of these two high altitude species.

Hypoxic adaptation of the hearts of plateau zokor (Myospalax baileyi) and plateau pika (Ochotona curzoniae) / 生理学报

Plateau zokor (Myospalax baileyi) and plateau pika (Ochotona curzniae) are native to the Qinghai-Tibet plateau. To study their adaptive mechanisms, the ratios of heart weight to body weight (HW/BW) and right to left ventricular plus septum weights [RV/(LV+S)] were determined; the microvessel density (MVD) of cardiac muscle were measured by immunohistochemical staining; the numerical density on area (N(A)), volume density (V(V)), specific surface (δ), and surface density (S(V)) of mitochondria were obtained by microscopy and stereology; the contents of myoglobin (Mb) and lactic acid (LD), and the activity of lactate dehydrogenase (LDH) in cardiac muscle were analyzed by spectrophotometer. The results showed that the HW/BW of plateau zokor [(4.55±0.26)%] and plateau pika [(4.41±0.38)%] was significantly greater than that of Sprague-Dawley (SD) rat [(3.44±0.41)%] (P<0.05), but the RV/(LV+S) [(22.04±1.98)%, (25.53±3.41)%] was smaller than that of SD rats [(44.23±3.87)%] (P<0.05). The MVD and N(A) of cardiac muscle were 1688.631±250.253 and 0.768±0.123 in SD rat, 2002.888±367.466 and 0.868±0.159 in plateau pika and 2 990.643±389.888 and 1.012±0.133 in plateau zokor. The V(V) of mitochondria in plateau zokor (0.272±0.045) was significantly lower than that in plateau pika (0.343±0.039) and SD rat (0.321±0.048) (P<0.05), while the δ of mitochondria in plateau zokor (9.409±1.238) was higher than that in plateau pika (6.772±0.892) and SD rat (7.287±1.373) (P<0.05). The S(V) of mitochondria in plateau pika (2.322±0.347) was not obviously different from that in plateau zokor (2.468±0.380) and SD rat (2.227±0.377), but that in plateau zokor was significantly higher than that in SD rat (P<0.05). The contents of Mb in cardiac muscle of plateau zokor [(763.33±88.73) nmol/g] and plateau pika [(765.96±28.47) nmol/g] were significantly higher than that of SD rat [(492.38±72.14) nmol/g] (P<0.05), the content of LD in plateau zokor [(0.57±0.06) mmol/L] was obviously higher than that in plateau pika [(0.45±0.06) mmol/L] and SD rat [(0.48±0.02) mmol/L] (P<0.05), and the activity of LDH in plateau zokor [(16.90±2.00) U/mL] and plateau pika [(20.55±2.46) U/mL] were significantly lower than that in SD rat [(38.26±6.78) U/mL] (P<0.05). The percentage of LDH-H in cardiac muscle decreased in order in plateau zokor, plateau pika and SD rat. In conclusion, plateau zokor and plateau pika adapt better to hypoxia than SD rat by increasing the SV of mitochondria, MVD and content of Mb in the cardiac muscle. However, the parameters of mitochondria in the two high-altitude animals are different possibly because of the differences of habitats and habits.

The vasoactive material and pulmonary vascular structural remodeling in rats / 中国应用生理学杂志

<p><b>AIM</b>To investigate the role of vascular endothelial growth factor (VEGF) and endothelin-1 (ET-1) on pulmonary vascular structural remodeling in rats and pika.</p><p><b>METHODS</b>The Wistar rats which reside at 2 260 m were carried to 3 417 m. After they were fed 24 hours,2 weeks and 3 weeks respectively, the level of VEGF and ET-1 were measured using a kit by ELISA method. Pulmonary tissue was taken out to stain with elastica-Van Gieson. The amount of pulmonary arteries (< 100 microm) and the component ratio of MA, PMA,and NMA were calculated by using a light microscope. The ratio of right ventricle weight to left ventricle plus septum weight (RV/LV + S) were measured.</p><p><b>RESULTS</b>The ET-1 was significantly different in pika as compared with 24 h, 2 weeks, 3 weeks hypoxic rats (P < 0.01) respectively. The levels of VEGF in 2 weeks, 3 weeks rats were much higher than that of pika but no difference was found between pika and 24 h hypoxic rats. The ratio of MA, PMA obviously increased, and NMA decreased significantly, right ventricular hypertrophy was developed in differ groups of hypoxic rats.</p><p><b>CONCLUSION</b>The VEGF and ET-1 participate the muscularization of pulmonary vessels during hypoxia and play an important role in the process of hypoxic pulmonary hypertension in rats, however the VEGF and ET-1 may be maintainable only normal organic function in pika.</p>