Effect of YIGU capsule on IGF-I mRNA and protein expression in rat osteoblasts in vitro

AIM: The effects of YIGU capsule on proliferation and IGF-I mRNA protein expressions in osteoblasts were studied. METHODS: (1) Forty 12-month old Sprague-Dawley female rats were divided randomly into four groups (YIGU capsule high dose group, medium dose group and low dose group; saline group), the drug-containing serum and control serum were prepared. (2) The new-born Sprague-Dawley rat osteoblasts were cultured with different YIGU capsule drug-containing serum at different concentrations and different exposure time. MTT method was used to observe proliferation of osteoblasts. (3) RT-PCR method was used to measure the relative IGF-I mRNA levels and ELISA method was used to measure IGF-I secretion at different exposure time. (4) ELISA method was used to measure IGF-I secretion at different exposure time. RESULTS: (1) Proliferation of osteoblasts was more than the control groups after 48, 72 and 96 h, respectively (P<0.01); (2) The relative IGF-I mRNA levels and IGF-I protein expression were higher than those in control group after 48, 72 and 96 h, respectively (P<0.01 or P<0.05). CONCLUSIONS: It was suggested that YIGU capsule drug-containing serum promoted proliferation, IGF-I mRNA and protein expression. These results may be parts of the mechanisms of YIGU capsule to prevent and treat osteoporosis.     

猪卵泡卵母细胞体外成熟与冷冻保存

研究了激素、猪卵泡液、不同类型血清对猪卵泡卵母细胞体外成熟的影响 ;比较了卵泡直径小于 2 mm、2～ 5 mm和大于 5 mm的卵母细胞体外成熟能力的差异 ,并对不同发育阶段猪卵母细胞的冷冻保存进行了研究。结果表明 :猪卵母细胞体外培养 4 8h时 ,培养的前 2 4 h培养液中加入激素 ,后 2 4 h去掉激素 ,卵母细胞的 A级成熟率 (5 1.73% )和总成熟率 (83.2 5 % )最高 ,极显著高于前 2 4 h不加激素 ,后 2 4 h添加激素培养的成熟率 (P<0 .0 1) ;也显著高于不含激素的培养液连续培养 4 8h的成熟率 (P<0 .0 5 ) ;但与添加激素连续培养 4 8h的成熟率差异不显著 (P>0 .0 5 )。在体外成熟培养液中 ,添加 10 % (体积分数 ) ECS的成熟率 (72 .86 % )显著高于添加 10 % (体积分数 ) NCS的成熟率(6 2 .2 1% ) (P<0 .0 5 ) ,而添加 10 % p FF则抑制卵母细胞的体外成熟。随着卵泡直径的增大 ,卵母细胞体外成熟能力逐渐增强。采用程序冷冻保存方法 ,成熟卵母细胞的成活率 (35 .5 9% )显著高于培养前 (2 4 .6 4 % )和培养 2 4 h(2 3.36 % ) (P<0 .0 5 )。培养 2 4 h(77.2 2 % )和培养成熟 (72 .81% )的卵母细胞解冻后的形态完整率均显著高于培养前(5 3.2 4 % ) (P<0 .0 5 )     

Clostridium difficile diarrhea: infection control in horses.

C difficile has emerged as an important cause of diarrheic disease in horses. C difficile diarrhea is usually diagnosed in mature horses, mostly when they are treated with antimicrobials and hospitalized. It is important for clinicians at veterinary hospitals to have knowledge about the organism and the infection. To prevent C difficile diarrhea, judicious use of antimicrobials is important, as is minimizing different stress factors at the animal hospital or clinic. Infected horses must be isolated. Routine examination for C difficile and toxin A or B is recommended in horses with antibiotic-associated diarrhea. When treating foals for R equi pneumonia, it is important to avoid accidental ingestion of erythromycin by the dams. To reduce the number of environmental spores, thorough cleaning and surface disinfection of the animal hospital and clinic are important. Routine handwashing should be performed by all staff.     

Renal ultrasonographic evaluation in the oncilla (Leopardus tigrinus).

Ultrasonography is currently used in veterinary medicine to examine the anatomy and physiology of internal organs and to establish normal standards for exotic animal species. Specifically, the kidneys may be evaluated and measured in length, width, and height. The kidneys in 33 oncilla (Leopardus tigrinus), including 20 males and 13 females, were opportunistically located, characterized, and measured by ultrasound using a 7.5-MHz linear transducer at two different zoologic facilities in S?o Paulo, Brazil. Renal volume was calculated from these linear measurements. The mean linear measurements and volume of the right and left kidneys were not significantly different.     

Recognition of the Nor98 variant of scrapie in the Swedish sheep population.

Within the framework of the active surveillance for transmissible spongiform encephalopathies in sheep in Sweden, 4 cases of the atypical form of scrapie, Nor98, were identified during 2003. Nor98 is a recently recognized and poorly understood variant of scrapie, first described in Norway. The cases were positive by the rapid test (enzyme-linked immunosorbent assay). Immunohistochemical staining showed diffuse thin-granular staining of the cerebellar cortex. Western immunoblotting analysis of specimens of brain stem and cerebellum showed a light band of approximately 12 kDa. Typical scrapie was ruled out based on the confirmatory testing. The affected ewes were from 4 different flocks. They were between 7 and 9 years old. Two were of the ARQ/ARQ genotype, 1 ARR/ARQ, and 1 ARR/AHQ. Two ewes had shown ataxia, and the other 2 had no clinical signs. Whole-flock slaughter was applied, and testing of the flock mates did not reveal additional cases. Nor98 differs from typical scrapie in its epidemiology, frequency of genotypes of sheep affected, clinical signs, microscopic lesions, distribution of scrapie prion protein in the brain, and characteristics of the immunostaining and immunoblotting profiles.     

Evaluation of the cationic trypsinogen gene for potential mutations in miniature schnauzers with pancreatitis

The purpose of this study was to evaluate the cationic trypsinogen gene in miniature schnauzers for possible mutations. Genetic mutations have been linked with hereditary pancreatitis in humans. Four miniature schnauzers were selected on the basis of a clinical history of pancreatitis. One healthy miniature schnauzer and 1 healthy mixed breed canine were enrolled as controls. DNA was extracted from these canines using a commercial kit. Primers were designed to amplify the entire canine cationic trypsinogen cDNA sequence. A polymerase chain reaction (PCR) was performed and products were purified and sequenced. All sequences were then compared. The healthy control canine, a healthy miniature schnauzer, and the 4 miniature schnauzers with pancreatitis showed identical sequences of the cationic trypsinogen gene to the published sequence. We conclude that, in contrast to humans with hereditary pancreatitis, mutations of the cationic trypsinogen gene do not play a major role in the genesis of pancreatitis in the miniature schnauzer.     

An inexpensive sedimentation chamber for the preparation of cytologic specimens of cerebrospinal fluid.

An inexpensive sedimentation chamber to obtain cytologic specimens of cerebrospinal fluid (CSF) is described. The device, which has a total cost of about $5.00 can be built in few minutes. The device permits the cytologic study of specimens of CSF in clinics, where because of economic constraints, a cytocentrifuge is not available. The device permits the study of the CSF cells on either air-dried or wet smears even under field conditions, and the results obtained are consistent. Also, the device permits to retrieve the cell-free fluid for its use in chemical or immunologic procedures.     

Effects of surgical implantation of temperature dataloggers on reproduction of captive striped skunks (Mephitis mephitis).

A total of 20 (14 females, six males) captive striped skunks (Mephitis mephitis) with miniature temperature dataloggers implanted free in the abdominal cavity were examined for reproductive performance and pathology because of implants. Eleven of 12 female skunks reproduced successfully 45.9+/-3.7 days after surgery to remove implanted dataloggers. The pregnancy rate of 91.7% (11/12) was much higher than other captive studies and was comparable with that of wild skunks. Eight striped skunks (six males, two females) that were euthanatized and necropsied after having implants in the abdominal cavity for 5 mo showed no apparent pathology associated with the implant. Implantation of the intraperitoneal devices did not lead to complications in abdominal tissues. Neither implant nor surgery affected reproduction. We conclude that implanted dataloggers can safely be used in physiologic studies of striped skunks or possibly other small carnivores in captive or field studies.     

Adherence of Actinobacillus pleuropneumoniae serotype 1 to swine buccal epithelial cells involves fibronectin

The swine pathogen Actinobacillus pleuropneumoniae serotype 1 was investigated for its ability to adhere to swine, rat, and human buccal epithelial cells (BEC). The highest number of bacteria adhered was to swine BEC. This binding ability was affected by heating, extreme pH, treatment with sodium dodecyl sulfate, ethylenediamine tetraacetate, or periodate, and proteolysis, suggesting that cell-surface glycoproteins participate in adherence and that adherence is based mostly on ionic interactions. Mannose and swine fibronectin may play a direct role in this interaction. Convalescent-phase serum from naturally infected pigs inhibited the adhesion. There was a correlation between bacterial pathogenicity as well as host specificity and the capacity for adherence to swine BEC. Adhesion to swine BEC provides a convenient method to study in vitro the adherence of A. pleuropneumoniae and other pathogens of the pig respiratory tract.     

生长肥育猪骨骼肌注射表达pGRF基因质粒的效应研究

将猪的GRF基因表达质粒注射于猪的骨骼肌后,研究其促生长效应与机理。选用始重6.3kg的44头去势长白×太湖仔猪,分6组,采用2×3因子安排的完全随机区组设计,按6～10kg、10～20kg、20～50kg、50～100kg4个阶段饲养。4个饲养阶段的饲粮低蛋白水平分别是20.70%,17.90%,15.03%,13.00%;高蛋白水平分别是23.70%,20.90%,18.02%,16.00%。pGRF基因质粒注射剂量设0mg、0.5mg、1.0mg3个水平,于试验开始与试验猪体重达60kg时共注射基因质粒2次。测定各阶段日增重,饲料消耗量,耗料增重比以及30、70、100kg时血液中GH、GRF、IGF-I的浓度。100kg时屠宰进行胴体品质测定。结果表明:饲粮蛋白水平对各阶段及全期试验猪日增重均有显著影响(P<0.05或P<0.01),对50～100kg阶段与全期日采食量和耗料增重比有显著影响(P<0.05或P<0.01)。注射pGRF基因质粒对各阶段及全期日增重均有显著影响(P<0.05),对6～10kg、10～20kg、50～100kg3阶段及全期日采食量有显著影响(P<0.05),对6～10kg阶段、50～100kg阶段及全期耗料增重比有显著影响(P<0.05)。注射pGRF基因质粒对30kg及70kg体重时猪血液中GRF、GH、IGF-I浓度均有显著影响(P<0.05)。提高饲粮蛋白水平与注射pGRF基因质粒均可明显降低超声波测膘厚及屠宰测膘厚、增大眼肌面积(P<0.05)。