Abstract:Objective To investigate the effects of Fu-Yuan-Zai-Zao capsule, a novel compound drug of traditional Chinese medicine, on learning and memory ability and its mechanism related to oxidative stress induced by global brain ischemia. Methods The mouse model of global brain ischemia was established by bilateral common carotid arteries occlusion and reperfusion. The learning and memory ability was measured by Morris water maze and step down tests. Nissl staining was used to detect the pathological changes in the hippocampal neurons. The activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) were evaluated by xanthinoxidase assay and thiobarbituric acid method, respectively. Results The global brain ischemia and reperfusion induced an impairment of learning and memory function, a decrease in the number of neurons in the hippocampal CA1 region, a decline in SOD activity and an increase in MDA content in the cerebral cortex of mice. Intragastrical administration of Fu-Yuan-Zai-Zao capsule for 14 d after brain ischemic surgery significantly improved the learning and memory impairment, increased the number of neurons in the hippocampal CA1 region, elevated SOD activity and reduced MDA content in the cerebral cortex. Conclusions Fu-Yuan-Zai-Zao capsule ameliorates the learning and memory impairment through inhibiting oxidative stress induced by global brain ischemia in mice. The results suggest that Fu-Yuan-Zai-Zao capsule may have future application in the treatment of ischemic cerebrovascular diseases.

Abstract:Objective To determine the normal range of hematological and visceral weight parameters of F1 4-week and 8-week old, male and female transgenic CB6F1 mice. The influence of gender and week age on the biochemical parameters was assessed. Methods 4-week and 8-week old CB6F1 mice, half male and half female (n=20 in each group), were weighed alive, dissected to weigh the main viscera, and blood samples were collected to test the physiological and biochemical parameters. Results When 4-week old and 8-week old CB6F1 mice were compared, there were significant differences in 22 parameters (body weight, heart, liver, spleen, left ovary, left testis, right testis, WBC, RBC, HGB, HCT, MCV, MCH, PCT, MPV, PDW, LYM, TP, ALT, ALB, P and TG) (P<0.01 for all), and in 8 parameters (left kidney, right kidney, right adrenal, thymus, left ovary, RDW, MON% and BUN) (P<0.05 for all). When male and female 4-week CB6F1 mice were compared, there were significant differences in 14 parameters (body weight, heart, liver, spleen, lung, left kidney, right kidney, MCHC, LYM, ALT, ALP, GLU, P and CHO) (P<0.01 for all), and in 6 parameters (right adrenal, WBC, PCT, MPV, TP and BUN) (P<0.05 for all). For male and female 8-week old CB6F1 mice, there were significant differences in 15 parameters (body weight, heart, liver, lung, left kidney, right kidney, MCV, PCT, LYM, LYM%, NEUT%, ALT, GLU, P and CHO) (P<0.01 for all), and in 5 parameters (WBC, RBC, MPV, NEUT and TP) (P<0.05 for all). Conclusions The normal range of hematological and visceral weight parameters of 4-week and 8-week old male and female CB6F1 mice are determined. Our study establishes normal detection indexes of CB6F1 mice and provides useful reference for its application.

Abstract:Objective To investigate the effects of dried whey on the intestinal bacterial community and probiotics in weaned laboratory rabbits. Methods A single factor design was employed to investigate the effects of dried whey supplemented at levels of 0%, 2%, 5% and 10%, respectively, on 48 weaned (40-day-old) laboratory rabbits. At the day 30, eight rabbits in each group were taken and sacrificed after anesthesia. The total bacterial DNA from the ceacal content of each selected rabbit was drew to analyze the bacterial community and intestinal probiotics (Bifidobacterium and Lactobacillius) population by PCR-DGGE and real-time fluorescence quantitative PCR, respectively. Results 1) The DGGE parameters of ceacal bacterial community were increased with the increasing dried whey supplemental levels. The number of DGGE band in 2%, 5% and 10% dried whey supplement groups (P<0.05, P<0.01), the Shannon index in 5% and 10% supplement groups (P<0.01) were significantly higher than that in the 0% supplement group, but the indices of DGGE band and Shannon index had no significant differences among the 2%, 5% and 10% dried whey supplement groups (P>0.05). Supplying dried whey has no significant effects on the homogeneity index (P>0.05). 2) The population of Bifidobacterium and Lactobacillius in ceacal content had a trend of increase with the rising dried whey supplement levels. Compared with the 0% supplement group, the Lactobacillius population in the 2%, 5% and 10% supplement groups (P<0.05), the Bifidobacterium population in the 10% supplement group (P<0.05) were significantly increased. Conclusions The results of our study indicate that: 1) Supplying dried whey in the feed of laboratory rabbit can effectively increase the diversity of ceacal bacterial community. 2) Dried whey may effectively improve the intestinal probiotics population.

Abstract:Objective Beagle dogs are commonly used animal for drug safety evaluation. As the necessary parameters, blood biochemical indicators are detected in acute or chronic toxicity tests. This study aims at assessing the influence of different preservation conditions and different preservation time on blood biochemical indicators to ensure the reliability of test results of long-term toxicity assessment.Methods Six Beagle dogs (3 males and 3 females) were used in this study. After collection and preparation of serum samples, biochemical indicators were detected after preservation in refrigerator at 2-8℃ for 1, 2, 5, 8, and 12 hours; after preservation in ice transportation boxes at 2-10℃ for 2, 5, and 8 hours; and after preservation in refrigerator at-20℃ for 1, 3, and 5 days. The biochemical indicators included alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP), total protein (TP), albumin (propagated), urea, creatinine (CREA), glucose (GLU), total cholesterol (TCHO), total bilirubin (TBIL), creatine kinase (CK), gamma pancreatic acyl transferase (GGT), calcium (CA), lactate dehydrogenase (LDH), phosphorus (P), high-density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), triglyceride (TG), sodium (Na⁺), potassium (K⁺) and chloride (Cl^-). Results Compared with the results of samples preserved for 1 hour, the LDL-C result of that preserved in refrigerator at-20℃ for 5 days was significantly increased (P<0.05), and the coefficient of variation was 39%. The CK and LDH results of specimens preserved in refrigerator at-20℃ for 5 days was increased but not significantly (P>0.05), and the coefficient of variation of LDH was 41%. Conclusions According to the test results of blood biochemical indicators in the Beagle dogs detected after different preservation conditions and different preservation time in this study, detection test should be done within 1 hour, if not, detection should be done within 12 hours for the samples preserved at 2~8℃, or within 3 days for the sample preserved at-20℃. For transportation of serum samples, the serum samples should be placed in the ice box at 2~10℃, and detection test should be done within 8 hours.

Abstract:Objective To compare the efficiency of bacteria culture and PCR assays for detection of Staphylococcus aureus (S. aureus), Pseudomonas aeruginosa (P. aeruginosa) and Klebsiella pneumoniae (K. pneumoniae) in laboratory rats and mice. Methods Bacteria culture combined with biochemical identification and PCR assay were used to detect 78 SPF rats and 422 SPF mice and the results of the two methods were compared. Results All the 78 rats were negative. Of the 422 mice, the positive rate by culture was 7.11% (30/422), of which, 10 were S. aureus, 22 were P. aeruginosa, and 2 were K. pneumoniae. The positive rate by PCR was 7.58% (32/422), of which, 10 were S. aureus, 25 were P. aeruginosa, and 2 were K. pneumoniae. Conclusions The high sensitivity, rapid procedure and easy to operate of PCR assay makes it valuable for rapid bacteria diagnosis and large-scale screening in laboratory animals.

Abstract:Objectives To explore the relativity between Alzheimer's disease (AD)-like lesions and metabolic syndrome models induced by high-sugar high-fat diet in rats. Methods Forty-eight Sprague Dawley rats were randomly divided into 2 groups.The control group (fed with normal diet, 12 rats) and high sugar and high fat group (fed with high-sucrose and high-fat diet, 12 rats) continuously for 12 months. At the end of 6, 9 and 12 months of the experiment, we observed the animal body weight and visceral fat weight. The blood lipid levels, blood glucose and MS-related biochemical parameters were determined. The brain tissues were examined by histopathology. The characteristic AD molecules hippocampus Aβ and Tau were detected using ELISA and Western blotting to confirm the presence of AD lesions in the brain. Results Compared with the normal control group, the body weight and visceral fat weight of the rats in the high-sugar high-fat groups were significantly increased; the levels of TG, FPG, LDL, HOMA-IR and hippocampus Aβ, phosphorylated Tau (p-Tau) were higher, but the level of HDL was decreased (P<0.05 for all).The histopathological examination revealed inflammatory cell infiltration in the brain tissues. Conclusions Characteristic AD-like lesions may occur and accompany the rat models of metabolic syndrome, induced by high-sugar high-fat diet, and provide a new idea for the construction of Alzheimer's disease animal models.

Abstract:Objectives To evaluate the effect of a Chinese traditional medicine, Huoxue Xiaoyi Fang, on the embryo development after in vitro fertilization (IVF) in mouse models of endometriosis. Methods To observe the quantity of oocytes including the average number of superovulation, fertility rate, 2-cell and 4-cell embryo cleavage rates at different stages in vitro fertilization, and morulation rate among the traditional Chinese medicine group (TCM group), endometriosis model group (EM model group) and sham-operated group. To explore the influence of endometriosis and the effect of Huoxue Xiaoyi Fang on the quantity of oocytes and embryonic development after IVF in the mouse models of endometriosis. Results All the average number of superovulation, fertility rate, 2-cell and 4-cell embryo cleavage rates in the three groups showed significant differences (P<0.05), and the average number of superovulation was in an order of sham operated group>TCM group>model group. There were no significant differences in the morulation rates in the three groups (P>0.05). There were significant differences among the IVF rates of the three groups (P<0.05), and to be in an order of sham operated group>TCM group>model group. There were significant differences in the final morulation rate among the three groups (P<0.05). Conclusions Endometriosis may influence the quality of embryonic development of EM model mice, including reduced average number of superovulation, fertility rate, and embryo cleavage rate. The Chinese traditional medicine Huoxue Xiaoyi Fang can improve the the quantity of embryonic development after IVF.

Abstract:Objective To accurately determine the biological characteristics, especially the disease-related indexes of SPF rabbits, and their gender differences. Methods A total of thirty 70-80 day old SPF New Zealand rabbits (male : female=1:1) were fed for a week, and then we weighed the body weight and main organs, determined the blood physiological and biochemical indexes, blood gas and arterial pressure, and measured the ventricular pressure by thoracotomy under respiratory support. Results Comparison of the male and female SPF New Zealand rabbits showed that there were significant differences in the mass of thyroid, adrenal gland, and liver (P<0.05 or P<0.01); the brain, pituitary gland, thyroid gland organ coefficients (P<0.05 or P<0.01); the thyroid/brain, adrenal/brain, and liver/brain ratios (P<0.05 or P<0.01); the mean erythrocyte volume and mean hemoglobin content (P<0.05 or P<0.01); and in glutamyl transpetidase and amylase (P<0.05 or P<0.01), but no significant differences in blood gas analysis, heart rate and carotid artery systolic and diastolic blood pressure, left ventricular systolic and diastolic blood pressure, and right ventricular systolic and diastolic blood pressure. Conclusions The gender has an impact on organ weight and blood physiological and biochemical indexes in New Zealand rabbits, but no significant influence on blood gas, blood pressure, heart rate and ventricular pressure in the rabbits.

Abstract:Objective To evaluate the diagnostic value of quantitative tissue velocity imaging (QTVI) in the detection of right ventricle function in a rabbit model of pulmonary artery embolism. Methods Twenty-two healthy adult rabbits were used in this study. The animal models of acute pulmonary embolism were created by injecting gelatin sponge into the ear vein of rabbits. The changes of right ventricle function were dynamically observed at immediately, 1 week, and 3 weeks after the establishment of acute pulmonary embolims models. QTVI images of the 4-chamber view at cardiac apex were recorded and analyzed by off-line analysis. Results The V_P was significantly reduced immediately after the models were built compared with the data before modeling in both the basal and middle segments, the TQ-S was prolonged, and Ve/Va>1 (P<0.05). The V_P at 1 week after modeling was not significantly decreased than the data before modeling in both the basal and middle segments, and the TQ-S was prolonged, but the Ve/Va ratio was <1 (P<0.05). At 3 weeks after modeling, the V_P was decreased compared with the data before modeling, and the Ve/Va ratio was <1 (P<0.05). The intra-group comparison showed that the V_P of the basal segment of right ventricle free wall was enhanced than that of the middle segment (P<0.05), but TQ-S was not obviously prolonged in both the two segments. Conclusions Quantitative tissue velocity imaging can reflect the contraction and relaxation of right ventricle in rabbits precisely. It is a good method to evaluate the function of right ventricle in animal models of pulmonary artery embolism.

Abstract:Objective To find the similarities and differences between SD rat and C57 mouse models of chronic renal failure established by 5/6 nephrectomy, and select an appropriate experimental animal for the study of the treatment and prevention of chronic renal failure. Methods 60 SD rats and C57 mice were randomly and equally divided into 5/6 nephrectomy group and sham operatied group. The mice and rats were killed at 4 weeks, 8 weeks and 12 weeks after the operation, respectively. Serum and renal tissue samples were collected for the detection and analysis of renal function and α-SMA. Results There was no significant difference in the survival rates between the rat and mouse models (P>0.05). The serum creatinine and urea nitrogen in the 5/6NX rat group showed a stable state at 8-12 weeks, which was different from that of the 5/6NX mouse group. Conclusions A 5/6 nephrectomy mouse model of chronic renal failure is successfully established. Although both 5/6 NX mouse and rat groups appear progressive renal failure after the operation, their disease processes are not quite the same. In clinic, appropriate experimental animals should be selected according to the length of the observation period in order to accurately show the disease states.

Abstract:Objective The aim of this study was to investigate the effect of left-handed pantoprazole sodium on rat models of gastric ulcer generated by pyloric ligation. Methods The rat models induced by pylorus ligation were treated with a single intravenous injection of left-handed pantoprazole sodium. The gastric ulcer inhibition rate, volume of gastric juice, activity of pepsin, and level of serum gastrin were determined and gastric mucosa tissue was examined by pathology. Results The gastric ulcer index, volume of gastric juice, activity of pepsin and level of serum gastrin in the rats pretreated with left-handed pantoprazole sodium (0.9, 1.8, 3.6 mg/kg, i.v.) were significantly lower than those in the model group. The lesions of gastric mucosa were also improved in the pantoprazole-treated groups. Conclusions Left-handed pantoprazole sodium has remarkable protective effects on the gastric mucosa in rat models of gastric ulcer induced by pyloric ligation.

Abstract:Objective To compare the advantages and values of several special staining methods of chondrocytes.Methods Twelve 7-day old healthy C57BL/6J mice were killed to obtain the cartilage tissue of the knee joint in order to isolate the chondrycytes. Type II collagen was used to assess the chondrocytes. Then the chondrocyte climbing slices were prepared. The materials were fixed, and HE staining, Safranin O-fast green staining, SA-β-gal staining and immunohistochemical staining of Type II collagen were performed and compared. Results HE staining showed clear morphology of the chondrocytes. The cell nuclei were stained blue and the cytoplasm was pink. Safranin O-fast green staining showed that the nuclei were pink and the cytoplasm green. SA-β-gal staining showed that the aging cells were green while the young cells were colorless. Immunohistochemical staining of type II collagen showed the distribution of type II collagen and they were stained brown while the cell nuclei were blue.Conclusions HE staining and safranin O-fast green staining can provide more information than the other staining techniques. SA-β-gal staining is useful in the analysis of aging chondrocytes. Immunohistochemical of type II collagen can be used to study type II collagen.

Abstract:Objective To establish a real-time quantitative PCR (qPCR) method for detection of Streptobacillus moniliformis, which can be used to rapidly detect this pathogen in laboratory animals. Method According to the S. moniliformis sequences published in NCBI, we designed specific primers and MGB probe. The specificity, sensitivity and stability of this method were evaluated using 24 standard reference strains. Total of 823 respiratory specimens of animals including mice, rats, guinea pigs, hamsters, rabbits, Mongolian gerbils and tree shrews, were detected by this established Taqman MGB qPCR method. Results We had successfully established the S. moniliformis Taqman MGB qPCR method. S. moniliformis was not detected in the samples of mice, rats, guinea pigs, hamsters and rabbits. The positive rate of S. moniliformis was 1.5% (1/65) and 61.7% (37/60) in conventional Mongolian Gerbils and tree shrews, respectively. Conclusions Our developed qPCR method can be used to effectively detect S. moniliformis in laboratory animals. Moreover, its accuracy and sensitivity are better than the national standard method. This study laid the foundations for optimizing the quality inspection system of laboratory animals.

Abstract:Autophagy is an important pathway in the body to remove damaged cell organelles and dysfunctional proteins to maintain protein equilibrium and intracellular homeostasis by the lysosomal system. As a degenerative disease of central nervous system, the main feature of Alzheimer's disease is abnormal protein aggregation. However, the role of autophagy in the pathogenesis of Alzheimer's disease is not fully clear yet. The aim of this paper is to take β-amyloid peptide as a starting point to explore the relationship between autophagy and β-amyloid peptide in the pathogenesis of Alzheimer's disease.

Abstract:Through years of experience to design, build, debug and operate the automatic control system in laboratory animal facilities, the author expounds the detailed effect of such system, analyzes the problems and causes, and finally provides some advice to improve the system.

Abstract:Objective To provide a practical device and protocol to hold conscious rats for subsequent operations which can overcome the disadvantages of existing methods. Users can complete the experiment more efficiently, with or without prior experience. Methods Using transparent plastic film, plastic sealing machine and sponge to make a simple device for holding rats, by taking advantage of their escaping nature. To compare the performance of the new method and existing methods for holding and injecting rats. Results Compared with existing methods, the new device and method can reduce the time-consuming to hold rats by 44.7%, from 18.13 seconds to 10.03 seconds. For holding and injecting, the new method can reduce the time-consuming by 55.3%, from 139.33 seconds to 52.26 seconds. Conclusions The new device and method is good for holding and injecting rats or drawing blood from the caudal veins. It can shorten the time of operation and reduce the stress reaction in the animals. It's especially helpful for inexperienced experimenters such as students in teaching and research tasks.

Abstract:China's scientific research and development sector are currently the second largest in the world, and laboratory animals are widely used across a range of research fields. As China's biomedical and pharmaceutical industry has grown, a laboratory animal science community has developed which now has its own specialist professional society-The Chinese Association for Laboratory Animal Sciences (CALAS). The importance of animal welfare is increasingly acknowledged in China and the value of, and need to provide, environmental enrichment is being recognized by more and more scientists. This article introduces the current understanding about and use of environmental enrichment for laboratory animals in China, and discussed the possible routes to develop environmental enrichment for laboratory animals in China.