Abstract:OBJECTIVE To investigate whether limb ischemic preconditioning (LIP) pretreatment has protective effects to the brain after cerebral ischemic injury by investigating the effects of LIP pretreatment on anti-inflammatory response in rats. METHODS Seventy-two Sprague Dawley rats were divided into 3 groups: experimental group (n=30), ischemic group (n=30) and control group (n=12). Six samples from five timepoints (6h, 12h, 24h, 48h and 72h after surgical manipulation) were collected for the experimental and ischemic groups, while other twelve animals were untreated and used as controls. The models with middle cerebral artery occlusion (MCAO) were obtained by thread blocking and the models with cerebral ischemic tolerance (IT) were obtained by LIP pretreatment. The determination of histopathological changes, real-time quantitative PCR (QTR-PCR) and enzyme-linked immunosorbent assay (ELISA) were used to observe the expression of interleukin-17 (IL-17) and interleukin-6 (IL-6). RESULTS The pathological change in experimental group was not as obvious as ischemia group. The results of QRT-PCR demonstrated that the expression of IL-17 and IL-6 were reduced significantly in experimental group. Significant different in the expression of IL-17 and IL-6 were found between ischemia and experimental groups at 12h、24h and 48h time points (P<0.01). The data of ELISA suggested that the expression of IL-17 and IL-6 were reduced obviously in experimental group. Significant different in the expression of IL-17 were found between ischemia and experimental groups at 12h、24h and 48h time points (P<0.05), while significant different in the expression of IL-6 were at 24h and 48h time points (P<0.05). CONCLUSION LIP pretreatment could induce ischemic tolerance, probably by providing obviously anti-inflammatory effects and protective function for the followed ischemic injury.

Abstract:As an efficient way, introduction of disease mouse models could provide support for scientific research in China. The practices about conservation and breeding of disease mouse models were reviewed in the article. It will be useful to guarantee the quality of imported disease mouse models.

Abstract:[Abstract] Objective To develop a RT-PCR method for determination of Mammalian Orthoreovirus 3 (Reo3) in laboratory animal and the biological materials or the biological products for human use from the animal. Methods Two pair of primers specific to the sequences of M1 gene were designed according to the published Mammalian Orthoreovirus 3.With which the RNA of Reo3 was extracted and reversely transcribed to cDNA as a template for PCR amplification.The developed RT-PCR method was optimized, verified for specificity and sensitivity. Results The developed RT-PCR method is good in ensitivity, specificity and stability, and its minimum detection limit using the recombinant plasmid containing Reo3 gene as atemplate was 0.42pg/μl,and the lowest detection virus titer is 10-9.ml-1. Conclusion The developed RT-PCR method can be used in detecting the Mammalian Orthoreovirus 3 (Reo3) in laboratory animaland the biological materials or the biological products for human use from the animal.

Abstract:Objective: The objective of this study is to observe the effect of photochemical ischemic stroke model made by hand-held laser in rats. Methods: 36 rats were randomly divided into four groups: group A, a skull hole was made deep to dura, irradiated by hand-held laser without rose-bengal injection for 5 minutes; group B, a skull hole was made deep to dura, irradiated by hand-held laser for 5 minutes with rose-bengal injection; group C, a skull hole was made deep to inner plate, irradiated by hand-held laser for 5 minutes with rose-bengal injection; group D, a skull hole was made deep to dura, irradiated by cold light source for 40 minutes with rose-bengal injection. Neurological deficit was evaluated 24 and 48 hours after surgery. MR scan was performed at 24 hours and the rats were killed at 48 hours, then the infarct volume was measured by TTC staining and the pathological changes was observed by light microscopy, comparing the success rate between two different irradiation sources. Results: The neurological deficit differed at 24 hours but converged at 48 hours. MR scan showed no infarction in group A and some in group D, obvious infarction in all rats from group B and C, but epidural hematoma occured in two rats of group C. TTC staining showed no infarction in group A, an infarction lesion occurred constantly in the left irradiated region in group B and C, but infarction ocurred only in half of the rats in group D. The successful rate of model-made was 100%、80% and 50% in group B, group C and group D respectively. Combining group B and group C into hand-held laser group, compared with the cold light source group (group D), there was a significant difference between two groups (P=0.026). Conclusions: In the procedure of photochemical ischemic stroke model made, compared with cold light source, the successful rate was higher when using hand-held laser as the irradiating source.

Abstract:Objective To design a kind of individual ventilated cage for breeding tree shrews on infectious diseases experimental purpose. Methods The cage was designed for tree shrews according to its biological characteristics , biosafety requirement and considering the cages of other experimental animals. Results The individual ventilated cage applies to use and operate in infectious diseases tree shrews model. Conclusion The individual ventilated cage can meet requirements of maintaining Animal welfare, ensuring animal quality, protecting person health and the environment against infection pollution. It has broad popularizing prospect to carry out infectious disease experiment.

Abstract:Objective Compare with the immunomodulatory function by Hirsutella sinensis and Paecilomyces hepiali in vivo. Method After the mice were administrated by Hirsutella sinensis and Paecilomyces hepiali.Observed carbon particle clearance function, delayed type hypersensitivity, hemolytic antibody, lymphocyte proliferation and NK cell activity in mice, and Cordyceps sinensis as a positive control. Result 3.0g·kg-1 Hirsutella sinensis and Paecilomyces hepiali can significantly improve the phagocyte index and half of hemolysis value (P<0.01). 3.0g·kg-1 Hirsutella sinensis and Paecilomyces hepiali can significantly improve the T, B lymphocyte proliferation (P<0.01, P<0.05). 3.0g·kg-1 Hirsutella sinensis can significantly enhance the activity of NK cells (P<0.05). 0.5-1.5g·kg-1 Hirsutella sinensis can significantly inhibit the ear edema in mice (P<0.01, P<0.05), at the same time 1.5g·kg-1 Hirsutella sinensis can inhibit the spleen and thymus increases(P<0.01, P<0.05).Conclusion Hirsutella sinensis and Paecilomyces hepiali have Immunomodulatory effects. Hirsutella sinensis.The Hirsutella sinensis were better than Paecilomyces hepiali in immunosuppression and enhance the role of the innate immune system.Paecilomyces hepiali were better than the Hirsutella sinensis in enhanced adaptive immune system. Keyword: Hirsutella sinensis; Paecilomyces hepiali; mice; Immunomodulatory function.

Abstract:【Abstract】 Objective Establish Rhesus monkey model of hyperlipidemia to provide reliable animal model for the study of human hyperlipidemia. Methods Designed high fat experimental feed formulations and detected its nutrients to feed Rhesus monkey to arouse hyperlipidemias. 2 mouths、4 mouths、6mouths later, we monitored the body weight, serum total cholesterol( TC ) , triglyceride ( TG ) , high-density lipoprotein cholesterol (HDL-C ), low density lipoprotein cholesterol (LDL-C),and analyzed the trend of each index above. Results When it was 2 moths and 4 months later,the feed intake and energy intake of model group were lower than control group（P<0.05）, but the body weight of model group was not significantly different from that of the control group( P > 0.05 );When it was 6 moths, the feed intake and energy intake of model group was not significantly different from that of the control group, and the body weight of model group were significantly different from that of the control group（P<0.05);When it was 2 moths later ,the indexes of TC,TG,HDL-C and LDL-C of the model group were not significantly different from that of the control group ( P > 0.05 ).When it was 4 moths later ,the indexes of TC,TG, LDL-C of the model group were significantly different from that of the control group （P<0.05). When it was 6 moths later ,the indexes of TC,LDL-C of the model group were significantly different from that of the control group（P<0.05),and much higher than the normal levels ,but the indexes of TG,HDL-C were not significantly different from that of the control group ( P > 0.05 ).According to the monitoring , TC and LDL-C showed a increase trend, TG showed a gradual recovery after the first descending trend ,and the HDL-C was not significantly changed . Conclusion Rhesus monkeys of the model group has a feature of high cholesterol and high low-density lipoprotein cholesterol. The paper has initialy established Rhesus monkey model of experimental hyperlipidemia.

Abstract:The irritable bowel syndrome (IBS) is a common disorder of unknown etiology. A group of dysregulated microRNAs (miRNAs) in blood microvesicles and in colon tissue have been identified in IBS patients. Research on miRNA, first phase may serve as a new way to further understand the pathophysiology of IBS characteristics, and eventually will leads us to use new strategy for diagnosis and treatment of IBS method by using miRNAs. Currently, there is not a lot of study with the role of miRNA in IBS. This paper summarize the work about miRNA on IBS.

Abstract:Objective To study the influence of Cramp gene knockout on the function of hematopoietic stem cell in aging. Methods Cells from the bone marrow of different age Cramp knockout and wild type mice were analyzed by flow cytometry. Results Compared with 12 months old wild type mice, percentage of long term hematopoietic stem cell decreased, multipotent hematopoietic progenitor increased; percentages of pre B and immature B cells decreased, and mature B cells increased in Cramp knockout mice. Conclusion Cramp gene knockout plays an important role in hematopoietic stem cell and the development of B lymphocytes in aging.

Abstract:Objective This study was designed to establish a simple and rapid methods for observing animal models of echinococcosis qualitatively and quantitatively. Methods Multilocular echinococcus in a dose of 2000 protoscolex / mL were intraperitoneally inoculated in the C. migratorus. Using a simple visual observation and microscopic examination method, the growth of intraperioneal cyst, cyst fluid and protoscoleces was observed in the 10d,15d, 18d, 22d, 39d and 60d after inoculation.. Results Protoscoles were found in the 18d, and cysts became big and many in the 15d,18d and 22d. There was protoscoles blastemal growth in the 39d. Different degree of protoscolex was developed in the 60d. With the extension of the inoculation time, the cyst weight increased. Conclusion The methods for examination of animal models of echinococcosis were established..

Abstract:Objective To develop a PCR for the detection of corynebacterium kutscheri （C. kutscheri） and apply it to clinical samples. Methods Genomic DNA was extracted as template for PCR from C. kutscheri (ATCC 11306) recoveried and cultivated in brain heart infusion medium. According to the C. kutscheri 16S rRNA gene sequence available in GenBank a pair of primes were designed and synthesized in order to develop a PCR for detection of C. kutscheri. After evaluated for sensitivity and specificity the PCR method was applied to detect the C. kutscheri in clinical livers and kidneys of mice artificially infected with C. kutscheri. Results The PCR for the detection of C. kutscheri was developed successfully and were specific enough to distinguish C. kutscheri from salmonella, streptococcus pneumoniae and Pasteurella. A minimum of 100 positive plasmids could be detected, indicating a good sensitivity of the assay. Conclusion This method repoted here is specific, sensitive and provides a fast detection of C. kutscheri and could be used for C. kutscheri clinical diagnosis.

Abstract:【Abstract】Objective To propose the formation of salt-sensitive hypertension and heart cell damage. Methods: Wistar rats, capsaicin was injected and high salt diet was feeded, the model of salt-sensitive hypertension was established. the change of heart histopathology were observed, spectrophotometry to detect heart iNOS activity and NO content; immunohistochemistry was performed to detect the expression of eNOS and iNOS protein in heart, the expression of eNOS and iNOS mRNA in heart was determined by RT-PCR, the level of apoptosis was assessed by single cell gel electrophoresis. Results: The rats body weight was not significant difference among groups at the end of the experiment (P>0.05). The capsaicin-high salt diet group systolic blood pressure was significantly higher than the other three groups (P<0.05) in 2, 3, 4 weeks. Capsaicin-high salt diet group cardiac muscle cell hypertrophy, muscle fiber disarrangement, nuclei arranged in irregular. The activity of iNOS and level of NO in capsaicin-high salt diet group significantly were higher when compared with control group. eNOS protein and eNOS mRNA significantly decreased compared with control group (P <0.01) iNOS protein and iNOS mRNA of capsaicin-high salt diet group were significantly increased compared with control group (P <0.01) in heart. The number of apoptotic cells of capsaicin-high salt diet group significantly increased compared with control group (P <0.05). Conclusions: The decreased express of eNOS mRNA and protein was related with the forming of salt-sensitive hypertension. The increased express of iNOS mRNA and protein could produce a large number of NO in heart. NO may make the cell apoptosis increasing and the damage of heart in salt-sensitive hypertensive rats.

Abstract:Objective To isolate, culture and identify the pathogenic bacterium from 6 cases of bacterial pneumonia within 1 month in tree shrew. Methods We Dissected the dead tree shrew ,cut open the lung tissue with sterile blade to insert focus of infection in the lung with inoculating loop to get samples for inoculating on nutrient agar and another two samples for bacterial smear with Gram staining and acid-fast staining. The bacterium obtained was isolated and cultivated for bacterial growth observation, then the pathogenic bacterium of tree shrew and its antibiotic susceptibilities was initially identified by Gram staining, acid-fast staining, oxidase test, biochemistrical encoding Identification system and 9-kind antibiotic susceptibility test. Results A amount of negative bacteria were observed from samples with Gram staining. Acid-fast staining appears it was nontuberculous mycobacterium about size 0.2μm?～6μm. Only 1 strain strong bacterium was observed in every 6 case on nutrient agar plate, it was verified as Gram-negative bacterium and nontuberculous mycobacterium, its size and staining results were the same as samples. It was identified as enteropathogenic E.coli. Antibiotic susceptibility test shows it is highly susceptibility to cefoperazone, furantoin, ampicillin, amikacin, ofloxacin, norfloxacin, sulfonamide methoxybenzyldine and low susceptibility to gentamicin and penicilin. Conclusion We conclude that Bacterial pneumonia is the cause of these 6 tree shrew death cases, the enteropathogenic E.coli.was initially identified as the reason. Drugs selected by antibiotic susceptibility could instruct clinical treatment like these cases in tree shrews.

Abstract:A ninety-day feeding trial was carried out to investigate the effect of protein on growth and serum biochemical Indexes of juvenile Tibetan macaque. Diets with five levels of protein (11.1%, 16.1%, 20.5%, 24.5% and 29.8%) were fed to Tibetan macaque. The results showed that: weight gain, serum albumin, globulin increased with the increasing protein levels (P < 0.05). Activities of serum aspartate aminortransferase, alanine aminotransferase and urea nitrogen were significantly improved in response to the increasing levels of dietary protein. But there was no significant difference in serum total protein, triglycerides, Gamma-irradiation glutamine shift enzyme and alkaline phosphates among these groups. The dietary protein requirement for weight gain of juvenile Tibetan macaque pestimated by brokenline model was 23.69%.

Abstract:Allergic asthma is an important disease. The interflow of asthma between animal models and human is that the former is also established on type I hypersensitivity. Sensitization is the first step for animal models of asthma. The characteristics and reactivity of complete allergens and haptens may lead to different outcomes. Airway challenge is a useful tool to study asthmatic responses. The new methods enabled successful observation of early-phase and late-phase asthmatic responses. Work on the pathogenesis and therapy of asthma should be conducted with different models besides the one of airway hyperresponsiveness used popularly nowadays. Other applications using animal models including airway remodeling, gene knock-out, transgenesis and therapeutic drugs were also reviewed briefly. 

Abstract:ObjectiveTo investigate whether limb ischemic preconditioning (LIP) pretreatment has protective effects to the brain after cerebral ischemic injury by investigating the effects of LIP pretreatment on anti-inflammatory response in rats.MethodsSeventy-two Sprague Dawley rats were divided into 3 groups:experimental group (n=30), ischemic group (n=30) and control group (n=12). Six samples from five timepoints (6 h, 12 h, 24 h, 48 h and 72 h after surgical manipulation) were collected for the experimental and ischemic groups, while other twelve animals were untreated and used as controls. The models with middle cerebral artery occlusion (MCAO) were obtained by thread blocking and the models with cerebral ischemic tolerance (IT) were obtained by LIP pretreatment. The determination of histopathological changes, real-time quantitative PCR (QTR-PCR) and enzyme-linked immunosorbent assay (ELISA) were used to observe the expression of interleukin-17 (IL-17) and interleukin-6 (IL-6). ResultsThe pathological change in experimental group was not as obvious as ischemia group. The results of QRT-PCR demonstrated that the expression of IL-17 and IL-6 were reduced significantly in experimental group. Significant different in the expression of IL-17 and IL-6 were found between ischemia and experimental groups at 12 h、24 h and 48 h time points (P<0.01). The data of ELISA suggested that the expression of IL-17 and IL-6 were reduced obviously in experimental group. Significant different in the expression of IL-17 were found between ischemia and experimental groups at 12 h、24 h and 48h time points (P<0.05), while significant different in the expression of IL-6 were at 24 h and 48 h time points (P<0.05). ConclusionLIP pretreatment could induce ischemic tolerance, probably by providing obviously anti-inflammatory effects and protective function for the followed ischemic injury.

Abstract:Objective To study the influence of Cramp gene knockout on the function of hematopoietic stem cell in aging. MethodsCells from the bone marrow of different age Cramp knockout and wild type mice were analyzed by flow cytometry. ResultsCompared with 12 months old wild type mice, percentage of long term hematopoietic stem cell decreased, multipotent hematopoietic progenitor increased; percentages of pre B and immature B cells decreased, and mature B cells increased in Cramp knockout mice. ConclusionCramp gene knockout plays an important role in hematopoietic stem cell and the development of B lymphocytes in aging.

Abstract:ObjectiveCompare with the immunomodulatory function by Hirsutella sinensis and Paecilomyces hepiali in vivo. MethodAfter the mice were administrated by Hirsutella sinensis and Paecilomyces hepiali.Observed carbon particle clearance function, delayed type hypersensitivity, hemolytic antibody, lymphocyte proliferation and NK cell activity in mice, and Cordyceps sinensis as a positive control. Result3.0g/kg Hirsutella sinensis and Paecilomyces hepiali can significantly improve the phagocyte index and half of hemolysis value (P<0.01). 3.0 g/kg Hirsutella sinensis and Paecilomyces hepiali can significantly improve the T, B lymphocyte proliferation (P<0.01, P<0.05). 3.0 g/kg Hirsutella sinensis can significantly enhance the activity of NK cells (P<0.05). 0.5～1.5 g/kg Hirsutella sinensis can significantly inhibit the ear edema in mice (P<0.01, P<0.05), at the same time 1.5 g/kg Hirsutella sinensis can inhibit the spleen and thymus increases(P<0.01, P<0.05). ConclusionHirsutella sinensis and Paecilomyces hepiali have Immunomodulatory effects. Hirsutella sinensis.The Hirsutella sinensis were better than Paecilomyces hepiali in immunosuppression and enhance the role of the innate immune system.Paecilomyces hepiali were better than the Hirsutella sinensis in enhanced adaptive immune system.

Abstract:Objective To generate transparent zebrafish, embryos may be treated with 1-phenyl 2-thiourea (PTU) to inhibit melanogenesis during embryogenesis. When working with techniques designed to study gene expression during embryogenesis, especially in teleosts, signal detection may be aggravated by pigmentation. Here we focus on the effects of PTU on embryo, and describes an optimized protocol for generating transparent zebrafish using PTU. Methods zebrafish embryos at the stage of 23 hpf(hours post fertilization) were treated with serial concentrations of PTU. Morphological and physiological endpoints were selected and quantified for unexposed and exposed embryos. Mortality, heart rate, hatching rate and SV-BA distance were detected until 80 hpf. ResultsExposure to 0.197mmol/L and 0.296 mmol/L PTU completely inhibited pigmentation while avoiding problems with embryo mortality, teratogenesis, reduced hatching frequency, and heart rate. The highest PTU concentration resulted in reduced hatching frequency and embryo mortality. We observed pericardial and yolk sac edema. Conclusion Optimized conditions can achieve complete inhibition of pigmentation for generating transparent zebrafish while avoiding toxic effects of PTU treatment.

Abstract:ObjectiveBy observing the influence of PKCβI membrane translocation and protein expression level in pulmonary artery Smooth muscle cells and Fibroblasts of chronic hypoxia-induced rat pulmonary hypertension, discuss the function of PKCβI in process of chronic hypoxia-induced rat pulmonary hypertension preliminary.MethodsChronic atmospheric pressure hypoxic PH model rats were established. Male SD rats were randomLy divided into six groups,and exposed to normoxia control or to normoxia hypoxia for 1d,3d,7d,14d and 21d to induce pulmonary hypertension. Western blot and immunohistochemistry were used to detect the expression of PKCβI membrane translocation and protein expression level in pulmonary artery Smooth muscle cells and Fibroblasts of chronic hypoxia-induced rat pulmonary hypertension. Results(1) RVSP and RV/(LV+S) increased significantly than normal control groups(P<0.05),rats pulmonary vascular obvious thickening after chronic hypoxia 3d,7d,14d and 21d. (2).PKCβI was expressed in rat pulmonary artery Smooth muscle cells and Fibroblasts, and its protein expressions decreased at 14d after chronic hypoxia than normal control groups(P<0.05).ConclusionsPKCβI protein expression is involved in the development of pulmonary vascular remodeling of chronic hypoxia-induced rat PH.

Abstract:Objective To establish Rhesus monkey model of hyperlipidemia to provide reliable animal model for the study of human hyperlipidemia. MethodsDesigned high fat experimental feed formulations and detected its nutrients to feed Rhesus monkey to arouse hyperlipidemias. 2 mouths、4 mouths、6mouths later, we monitored the body weight, serum total cholesterol( TC ), triglyceride ( TG ), high-density lipoprotein cholesterol (HDL-C ), low density lipoprotein cholesterol (LDL-C),and analyzed the trend of each index above. ResultsWhen it was 2 moths and 4 months later,the feed intake and energy intake of model group were lower than control group(P<0.05), but the body weight of model group was not significantly different from that of the control group( P > 0.05 );When it was 6 moths, the feed intake and energy intake of model group was not significantly different from that of the control group, and the body weight of model group were significantly different from that of the control group(P<0.05);When it was 2 moths later,the indexes of TC,TG,HDL-C and LDL-C of the model group were not significantly different from that of the control group ( P > 0.05 ).When it was 4 moths later,the indexes of TC,TG, LDL-C of the model group were significantly different from that of the control group(P<0.05). When it was 6 moths later,the indexes of TC,LDL-C of the model group were significantly different from that of the control group(P<0.05),and much higher than the normal levels,but the indexes of TG,HDL-C were not significantly different from that of the control group ( P > 0.05 ).According to the monitoring, TC and LDL-C showed a increase trend, TG showed a gradual recovery after the first descending trend,and the HDL-C was not significantly changed.ConclusionRhesus monkeys of the model group has a feature of high cholesterol and high low-density lipoprotein cholesterol. The paper has initialy established Rhesus monkey model of experimental hyperlipidemia.

Abstract:ObjectiveTo investigate the effect of protein on growth and serum biochemical indexes of juvenile Tibetan macaqueb by a ninety-day feeding trial. MethodDiets with five levels of protein (11.1%, 16.1%, 20.5%, 24.5% and 29.8%) were fed to Tibetan macaque. The results showed that:weight gain, serum albumin, globulin increased with the increasing protein levels (P<0.05). ResnltsActivities of serum aspartate aminortransferase, alanine aminotransferase and urea nitrogen were significantly improved in response to the increasing levels of dietary protein. But there was no significant difference in serum total protein, triglycerides, Gamma-irradiation glutamine shift enzyme and alkaline phosphates among these groups. The dietary protein requirement for weight gain of juvenile Tibetan macaque pestimated by brokenline model was 23.69%.ConclusimUnder the cage conditions the 23.69% protein is the best choice considering animal biology and feed expense.

Abstract:ObjectiveTo propose the formation of salt-sensitive hypertension and heart cell damage. MethodsWistar rats, capsaicin was injected and high salt diet was feeded, the model of salt-sensitive hypertension was established. the change of heart histopathology were observed, spectrophotometry to detect heart iNOS activity and NO content; immunohistochemistry was performed to detect the expression of eNOS and iNOS protein in heart, the expression of eNOS and iNOS mRNA in heart was determined by RT-PCR, the level of apoptosis was assessed by single cell gel electrophoresis. ResultsThe rats body weight was not significant difference among groups at the end of the experiment (P> 0.05). The capsaicin-high salt diet group systolic blood pressure was significantly higher than the other three groups (P<0.05) in 2,3, 4 weeks. Capsaicin-high salt diet group cardiac muscle cell hypertrophy, muscle fiber disarrangement, nuclei arranged in irregular. The activity of iNOS and level of NO in capsaicin-high salt diet group significantly were higher when compared with control group. eNOS protein and eNOS mRNA significantly decreased compared with control group (P<0.01) iNOS protein and iNOS mRNA of capsaicin-high salt diet group were significantly increased compared with control group (P<0.01) in heart. The number of apoptotic cells of capsaicin-high salt diet group significantly increased compared with control group (P<0.05). ConclusionsThe decreased express of eNOS mRNA and protein was related with the forming of salt-sensitive hypertension. The increased express of iNOS mRNA and protein could produce a large number of NO in heart. NO may make the cell apoptosis increasing and the damage of heart in salt-sensitive hypertensive rats.

Abstract:Objective To develop a RT-PCR method for determination of Mammalian Orthoreovirus 3 (Reo3) in laboratory animal and the biological materials or the biological products for human use from the animal. Methods Two pair of primers specific to the sequences of M1 gene were designed according to the published Mammalian Orthoreovirus 3.With which the RNA of Reo3 was extracted and reversely transcribed to cDNA as a template for PCR amplification.The developed RT-PCR method was optimized, verified for specificity and sensitivity. Results The developed RT-PCR method is good in ensitivity, specificity and stability, and its minimum detection limit using the recombinant plasmid containing Reo3 gene as atemplate was 0.42pg/μL,and the lowest detection virus titer is 10-9/mL. Conclusion The developed RT-PCR method can be used in detecting the Mammalian Orthoreovirus 3 (Reo3) in laboratory animaland the biological materials or the biological products for human use from the animal.

Abstract:ObjectiveTo develop a PCR for the detection of corynebacterium kutscheri(C. kutscheri)and apply it to clinical samples. Methods Genomic DNA was extracted as template for PCR from C. kutscheri (ATCC 11306) recoveried and cultivated in brain heart infusion medium. According to the C. kutscheri 16S rRNA gene sequence available in GenBank a pair of primes were designed and synthesized in order to develop a PCR for detection of C. kutscheri. After evaluated for sensitivity and specificity the PCR method was applied to detect the C. kutscheri in clinical livers and kidneys of mice artificially infected with C. kutscheri. ResultsThe PCR for the detection of C. kutscheri was developed successfully and were specific enough to distinguish C. kutscheri from salmonella, streptococcus pneumoniae and Pasteurella. A minimum of 100 positive plasmids could be detected, indicating a good sensitivity of the assay. Conclusion This method repoted here is specific, sensitive and provides a fast detection of C. kutscheri and could be used for C. kutscheri clinical diagnosis.

Abstract:ObjectiveTo explore the ideal experimental rats model for research on hyperlipidemia. Methods Three prescriptions on experimental hyperlipidemia-rats were modified and emulsion was made by references. At 14 day blood samples were taken from ocular orbit and content of TC、TG、LDL、HDL in the serum were measured. ResultsThree prescriptions could lead to the disorder of lipid metabolism at different levels. Conclusion Hyperlipidemia-rats model can be formed in a short time by prescription 3.

Abstract:ObjectiveThis study was designed to establish a simple and rapid methods for observing animal models of echinococcosis.MethodsMultilocular echinococcus in a dose of 2000 protoscolex/mL were intraperitoneally inoculated in the C. migratorus. Using a simple visual observation and microscopic examination method, the growth of intraperioneal cyst, cyst fluid and protoscoleces was observed in the 10d,15d, 18d, 22d, 39d and 60d after inoculation. ResultsProtoscoles were found in the 18d, and cysts became big and many in the 15d,18d and 22d. There was protoscoles blastemal growth in the 39d. Different degree of protoscolex was developed in the 60d. With the extension of the inoculation time, the cyst weight increased. Conclusion The methods for examination of animal models of echinococcosis were established.

Abstract:Objective The objective of this study is to observe the effect of photochemical ischemic stroke model made by hand-held laser in rats. Methods 36 rats were randomly divided into four groups:group A, a skull hole was made deep to dura, irradiated by hand-held laser without rose-bengal injection for 5 minutes; group B, a skull hole was made deep to dura, irradiated by hand-held laser for 5 minutes with rose-bengal injection; group C, a skull hole was made deep to inner plate, irradiated by hand-held laser for 5 minutes with rose-bengal injection; group D, a skull hole was made deep to dura, irradiated by cold light source for 40 minutes with rose-bengal injection. Neurological deficit was evaluated 24 and 48 hours after surgery. MR scan was performed at 24 hours and the rats were killed at 48 hours, then the infarct volume was measured by TTC staining and the pathological changes was observed by light microscopy, comparing the success rate between two different irradiation sources. ResultsThe neurological deficit differed at 24 hours but converged at 48 hours. MR scan showed no infarction in group A and some in group D, obvious infarction in all rats from group B and C, but epidural hematoma occured in two rats of group C. TTC staining showed no infarction in group A, an infarction lesion occurred constantly in the left irradiated region in group B and C, but infarction ocurred only in half of the rats in group D. The successful rate of model-made was 100%、80% and 50% in group B, group C and group D respectively. Combining group B and group C into hand-held laser group, compared with the cold light source group (group D), there was a significant difference between two groups (P=0.026). ConclusionsIn the procedure of photochemical ischemic stroke model made, compared with cold light source, the successful rate was higher when using hand-held laser as the irradiating source.

Abstract:The irritable bowel syndrome (IBS) is a common disorder of unknown etiology. A group of dysregulated microRNAs (miRNAs) in blood microvesicles and in colon tissue have been identified in IBS patients. Research on miRNA, first phase may serve as a new way to further understand the pathophysiology of IBS characteristics, and eventually will leads us to use new strategy for diagnosis and treatment of IBS method by using miRNAs. Currently, there is not a lot of study with the role of miRNA in IBS. This paper summarize the work about miRNA on IBS.

Abstract:Experimental rabbits,one of the most important experimental animals,played an imptant role in medical field. All kinds of genetic markers methods provided more simple, reliable research methods for system development, population genetic structure and quality control, and other fields. However, the application of domestic experimental rabbits is restricted because of their unstalbe quality and unclear genetic background. In this paper, applications of the technology of molecular markers on the analysis of genetic diversity of rabbits are summarized, aimed at providing some help to genetic testing of experimental rabbits.

Abstract:ObjectiveTo design a kind of individual ventilated cage for breeding tree shrews on infectious diseases experimental purpose. MethodsThe cage was designed for tree shrews according to its biological characteristics, biosafety requirement and considering the cages of other experimental animals. ResultsThe individual ventilated cage applies to use and operate in infectious diseases tree shrews model. ConclusionThe individual ventilated cage can meet requirements of maintaining animal welfare, ensuring animal quality, protecting person health and the environment against infection pollution. It has broad popularizing prospect to carry out infectious disease experiment.

Abstract:ObjectiveTo isolate, culture and identify the pathogenic bacterium from 6 cases of bacterial pneumonia within 1 month in tree shrews. MethodsWe Dissected the dead tree shrew, cut open the lung tissue with sterile blade to insert focus of infection in the lung with inoculating loop to get samples for inoculating on nutrient agar and another two samples for bacterial smear with Gram staining and acid-fast staining. The bacterium obtained was isolated and cultivated for bacterial growth observation, then the pathogenic bacterium of tree shrew and its antibiotic susceptibilities was initially identified by Gram staining, acid-fast staining, oxidase test, biochemistrical encoding identification system and 9-kind antibiotic susceptibility test. ResultsA amount of negative bacteria were observed from samples with Gram staining. Acid-fast staining appears it was nontuberculous mycobacterium about size 0.2 μm×2~6 μm. Only 1 strain strong bacterium was observed in every 6 case on nutrient agar plate, it was verified as Gram-negative bacterium and nontuberculous mycobacterium, its size and staining results were the same as samples. It was identified as enteropathogenic E.coli. Antibiotic susceptibility test shows it is highly susceptibility to cefoperazone, furantoin, ampicillin, amikacin, ofloxacin, norfloxacin, sulfonamide methoxybenzyldine and low susceptibility to gentamicin and penicilin. ConclusionWe conclude that Bacterial pneumonia is the cause of these 6 tree shrew death cases, the enteropathogenic E.coli.was initially identified as the reason. Drugs selected by antibiotic susceptibility could instruct clinical treatment like these cases in tree shrews.

Abstract:As an efficient way, introduction of disease mouse models could provide support for scientific research in China. The practices about conservation and breeding of disease mouse models were reviewed in the article. It will be useful to guarantee the quality of imported disease mouse models.