Abstract:Objective To compare the tumor growth and metastasis of three different mouse breast tumor cells with Luc-labeled (4T1-Luc, 66c14-Luc and 4TO7-Luc) by using bioluminescent imaging system, and to develop ideal tumor model and living animal analysis technique for tumor metastasis and anti-metastatic therapy. Method The vector containing luciferase gene was constructed and transfected into mouse breast tumor cell line cells and selected with G418 to obtain stable Luc-expressing clones. The cells in logarithmic phase was collected and diluted to 1?07 cells/mL. Then 0.1 mL cell suspension was inoculated into the second mammary fat pad on the right side of normal BALB/c mice to establish the orthotopic models. And another 0.1 mL cell suspension was intravenously inoculated into the tail vein of BALB/c mice to establish the tail vein xenografted model. Their tumorigenesis and metastasis were analyzed in vivo. Result The stable Luc-expressing cell lines were obtained. The whole-body optical imaging found that tumors could be formed after 7 days when the cells were inoculated orthotopically. After 28 days, the tumor sizes of 4T1 were the biggest, the tumor sizes of 66c14 were the second biggest and 4TO7 were the smallest. After 35 days, the tumor sizes were similar among three kinds of cells, while the tumor metastases were showed between 4T1 and 66c14 cells. The metastasis of 4T1 tumor was more serious than 66c14 tumor. 4TO7 tumors didn’t transfer. After 42 days, the tumor sizes were similar among three kinds of cells. The metastasis of 4T1 and 66c14 tumor became more seriously with time passed, and the metastasis of 4T1 tumor was more extensive than 66c14 tumor. 4TO7 tumors still didn’t transfer. After 7 days, the whole-body optical imaging found that tumor could be formed in lung when the stable Luc-expressing cell suspension was intravenously inoculated into the tail vein. The fluorescent signal of 4T1 tumor was the strongest, and deaths of mice occurred in succession. The fluorescent signal of 4TO7 tumor was the second strongest, and 66c14 was the weakest. After 14 days, the metastasis of 4TO7 and 66c14 tumor became more seriously with time passed, and the fluorescent signal of 4TO7 tumor was stronger than 66c14 tumor, and deaths of mice occurred in succession. Conclusions Breast spontaneously metastatic model could show the metastatic characteristics and the whole metastatic process more authentic than the tail vein xenografted model，which is an ideal model for the study of tumor metastasis.

Abstract:【Abstract】Objective To discuss the differences of Muc2 and DCN gene knockout mice on kronismus. Methods Knockout homozygote males and females (Muc2－/－,DCN－/－ ) were mated respectively according to 1:1 or 1:2. The 1-3 generation of average litter sizes ,parity time interval, and kronismus in Muc2 and DCN gene knockout mice were observed.Results An average litter sizes of Muc2 gene knockout mice are 5.80 + 0.95. the average birth time interval is 42.29 + 2.28 days; DCN knockout mice seed production on average 3.85 + 0.76, the average birth time interval is 24.86 + 10.42 days.Muc2 and DCN gene knockout mice in average litter sizes ,parity time interval, and kronismus are obviously different. Conclusions Two groups of knockout mice reproductive performance are different.It reveal may be associated with Muc2 and DCN gene.

Abstract:Objective To study the changes of liver, aortic wall and heart function in hyperlipidemia hamster at different time points via ultrasonic imaging, and investigate the value of ultrasonic diagnosis in hamster model of hyperlipidemia. Methods 32 Syrian Golden hamsters were randomly divided into two groups that were fed with normal chow (control group) and high fat diet (model group) respectively. Serum lipids levels were analyzed after 0, 3, 6, 9, 14, 16, 20, 24, 40 weeks using commercial kits. Hamsters from each group were examined with ultrasound and then sacrificed for pathological examination in the 20th week and 40th week. Results Serum levels of TC, TG, HDL-C and LDL-C increased remarkably in hamsters fed with high fat diet and maintained significantly above the normal levels throughout the study course (p<0.05). Compared with the control group, hamsters fed with high fat diet for 20 weeks had greater internal-media thickness (IMT) (p<0.0001) and their IMT increased significantly when the dietary intervention period was extended to 40 weeks (p<0.0001). It was revealed by both ultrasonic imaging and pathological examination that fatty liver and aortic arch atherosclerotic lesions were developed in hyperlipidemia hamsters fed with the high fat diet for 20 weeks, and aggravated during the prolonged 20 weeks. Conclusion IMT of aortic arch walls of hamster can be dynamically measured through ultrasonic imaging. It can also be used to monitor liver and sensitively detect the morphological changes of aortic arch walls of hyperlipidemia hamster.

Abstract:Objective To develop a detection method for antibiotic residues in laboratory animal serum, which ensure the accuracy of the results of the microbiological quality monitoring. Methods Antibiotic residues of 1640 laboratory animal sera were test by microbial inhibition assay of Bacillus stearothermophilus. And the results compared with the antibiotic detection kit Premi@Test. Results Antibiotic residues were positive in sera of mice, rats, guinea pigs, rabbits and dogs, the positive rate of 27.2%, 16.4%, 39.9%, 23.7% and 47.6%, respectively. The results were consistent with Premi@Test（X2=16.680，P>0.05). Sera antibiotic residues and DHL agar colony growth had certain correlation(X2=9.939, P <0.05). Conclusions The microbial inhibition method of Bacillus stearothermophilus used as indicator is not only capable of detecting antibiotic residues in animal sera, low cost, simple operation, but also has a good sensitivity and repeatability. This study provides a basis for detection of antibiotic residues in the laboratory animals, and makes up for the blank in the quality testing of laboratory animals.

Abstract:[Abstract] Objective To study the toxicity of rats that caused by long-term intake of low-dose Clethodim Technical and determine the maximal non-effect of Clethodim Technical, in order to provide the reference dose for safety in production and chronic toxicity experiment. Methods A total of 80 SD rats(half female and half male) were randomly divided into 4 groups, respectively, for the orginal drug 14.9mg/kg?bw?d group, 89.6mg/ kg?bw?d group, 537.5mg/ kg?bw?d and normal control group based on body weight. Execute the rats meanwhile detect the indexes including hemology、biochemistry detection of serum、body weight and organ module after the end of the experiment, and the results were analyzed statistically. Results and Conclusions When the oral dosees of Clethodim Technical at 200 mg/ kg?bw?d and above, it can cause toxic effects on rats, and its maximal non-effect dose of long-term intake of low-dose Clethodim Technical on rats is 40 mg/kg?bw?d.

Abstract:Objective:Based on the environmental testing results of the 2012 annual laboratory animal license in Hubei Province, we will discuss the results and make suggestions to promote the reliable operation of the experimental animal facility in Hubei Province.Methods:The environment was monitored in 30% of the regional sampling way on the basis of GB14925-2010 Laboratory animal.Result:In addition to the air velocity and maximum average concentration Settlement bacteria, other indexes don’t meet national standards..Conclusion:To strengthen Laboratory animal facility management and training and improve the level of management overall improve the environmental conditions of laboratory animal facility in Hubei Province.

Abstract:Object: There is growing concern about the role of monocytes-macrophages in the progress of atherosclerosis (AS). However, the mechanism of lipid change in those cells in the foaming process is not clear. Methods: LDL was isolated by density gradient centrifugation at one time and was made as Ox-LDL. Furthermore, the changes of lipid composition and cellular shape could be dynamically observed in rat macrophages. Results: the purity level of purified LDL can reach 92.39%. The increased red lipid droplets in macrophages were observed by the experimental method of cellular morphology. The electron microscopy showed that chromatin around nucleus , numerous mitochondria and ribosome in the sparse cytoplasm. In LDL group, with the increasing of LDL level , the TC, FC, CE were all increased .There was the same result in Ox-LDL group. Conclusion: Interaction of Ox-LDL and monocytes-macrophages could promote part of arterial wall to form the cells of AS feature. OX-LDL was easier to make monocytes/macrophages change to foam cells than LDL. Moreover, it was confirmed that high level of OX-LDL can lead to damage of cells membrane

Abstract:Premature Ovarian Failure (POF) is a classic heterogeneous disease with different causes. In recent years, studies show there has been a significant increase in its incidence. POF seriously affect physical and memtal health of women. Finding a perfect and reliable animal model plays an important role in POF research. This review summarized the relatively successful animal models established by researchers at home and abroad and discussed their advantages and disadvantages briefly.

Abstract:Objective To analyze the effects of blood-glucose fluctuation on inflammatory hippocampus damage in diabetes mellitus model of rats. Methods The diabetic rats model (M) was established by injection of streptozotocin (STZ), the persistent high glucose diabetes rats model group (MS) was established by intraperitoneal injection of glucose, and the blood-glucose fluctuation rats model group (MF) was developed by alternative intraperitoneal injection of insulin and glucose administration. Body weight, glucose (Glu), triglyceride (TG), low density lipoprotein (LDL-C）and high density lipoprotein (HDL-C) were measured at 6 weeks after MF model establishment. The mRNA expression levels of IL-1β, IL-2, IL-6, IL-8 and TNF-α in the rat hippocampus were detected by real-time PCR. The influence of blood-glucose fluctuation on learning and memory function of hippocampus was observed using of Morris water maze. Results (1) The body weight of M group, MS group and MF group were significantly decreased comparing to normal rats group (N) (P<0.01). (2) 1 week after intravenous injection of STZ, the levels of Glu, TG and LDL-C in M group, MS group and MF group were significantly increased (all P<0.01), and HDL-C was significantly decreased comparing to N group (P<0.01). (3) Comparing to N group, the expression levels of IL-1β、IL-6、IL-8 and TNF-α mRNA in M, MS and MF groups were all changed significantly (all P< 0.05), while there was no significant difference in the expression of IL-2 mRNA in all rats groups (P> 0.05). The expression levels of IL-1β and TNF-α mRNA in MF group were significantly higher than those of M and MS group (all P<0.01). (4) The escape latency, the number of rats passing hidden platform and the swimming distance in the quadrant of hidden platform of M, MS and MF groups were significantly higher than those of N group (all P<0.01). And the escape latency, the number of rats passing hidden platform in MF group were significantly higher than those of M and MS groups. While there was no significant difference in swimming distance in the quadrant of hidden platform among M, MS and MF groups (P> 0.05). Conclusion Blood glucose fluctuation could induce inflammatory damage and dysfunction of hippocampus in rats more seriously than that induced by acute hyperglycemia and chronic persistent hyperglycemia.

Abstract:Tree shrews is similar to human beings in biological characteristics, physiology and biochemistry, metabolism, genome and so on, it has played an important role in many scientific research fields, such as physiology, pathology, virology, immunology, pharmacology, genetics and so forth . To this day most of wild tree shrews were used in biomedical field, there is no national quality control standard. In order to assure standardization and scientific of tree shrews introduction, domestication, alimentation, reproduction, quality control, animal welfare, and increase reproduction rate and the survival rate of tree shrew. According to requirement of veterinary public health and laboratory animal sciences, this paper summarized some basic principles, technical operation and welfare requirements relate to tree shrews, which includes wild catching ,transportation, quarantine, feeding , breeding and others, it is beneficial to interrelated workers and correlation studies.

Abstract:Objective To observe the clinical efficacy of pioglitazone after intensive glycemic control in treating with type 2 diabetes. Method 60 patients suffering from type 2 diabetes previously treated with oral antihyperglycemic drugs with inadequate glycemic control first received continuous subcutaneous insulin injection. After plasma glucose levels achieved the target(FPG＜7.0mmol/L,2hPG＜10.0mmol/L), the treatment was changed to mealtime insulin aspart combining with bedtime insulin glargine. One week later, patients were randomized in a 1:1 ratio into two groups, one group only received insulin aspart and insulin glargine (control group), the other group took pioglitazone hydrochloride 30mg per day in addition to insulin aspart and glargine(treatment group). If FPG or 2hPG decreased too fast or a hypoglycemic reaction happened, the insulin dosage would be reduced. The observation period was 12 weeks. We observed the FPG, 2hPG, HbAlc, insulin dosage, triglyceride(TG), total cholesterol(TC), high density lipoprotein cholesterol(HDL-C), low density lipoprotein cholesterol(LDL-C) in these two groups before randomization and in week 12. Results 1、The insulin dosage decreased obviously in treatment group after the addition of pioglitazone(P＜0.05), but no significant difference was observed in control group(P＞0.05); 2、FPG and 2hPG did not changed significantly in both groups after 12 weeks of treatment. 3、HbA1c declined significantly in both groups after the treatment (P＜0.05), but no significant difference was observed between two groups(P＞0.05). 4、TG decreased and HDL-C increased significantly in treatment group after 12 weeks of treatment(P＜0.05)，while no significant difference was observed in control group, and the TG was significantly lower and HDL-C was significantly higher in treatment group than those in control group(P＜0.05). Conclusions After the patients had received the intensive insulin treatment and the plasma glucose has reached the goal. The addition of pioglitazone can significantly decrease the insulin dosage that patients need, improve patient compliance, and at the same time improve lipid metabolism, providing some benefits in reducing the occurrence of cardiovascular complications.

Abstract:Objective The experiment investigates the influence of the Low-Intensity Pulsed ultrasound Stimulation to restore the segment bone defect. Methods Insert 12mm diametric 20mm long TiC/Ti Foam into the area of Beagle dogs’ left tibia with segmental bone defect. Divide the six dogs into two groups randomly, the experiment group with LIPUS (1.5MHz, ISATA30mW/cm2, pulsed width 200μs, repeating 1kHz pulsation, once/d), the contra group without LIPUS. Observe and analyse formation around the material with X-ray film and the admeasurement of the bone mineral density at 4w and 8w. Results Six Beagle dogs were included in the final analysis. At week 4, the maturation of newly formed bone under the LIPUS treatment group appeared earlier than that of the contra group, on the basis of the bone mineral density increase and bone callus growth. On the bone mineral density aspect, the experimental group’s rate is higher than the contra group; on the bone callus aspect, the experimental group grow from marginal to central, the contra group still has vacancy. At week 8, the skeletal structure stability under the LIPUS treatment group is higher than that of the contra group, on the basis of the bone callus area and the formability. Measured the bone mineral density at week 4, the experimental group is higher than the contra group(p<0.05). Conclusion Combining applying the Low-Intensity Pulsed Ultrasound Stimulation and the artificial bone material could enhance the formation rate of the bone callus and the bone mineral density, and shorten the healing time of the segmental bone defect.

Abstract:Objective ultrasonography was used to estimate alcoholic fatty liver model in rats. Methods Forty healthy male SD rats were selected randomly and divided into two groups(n=20). According to weight measured weekly, the rats in model group were gavaged with ethanol(2 times/day). The ethanol concentration was 40% at the first week, then was increased to 45% and 50% at the second and third week, respectively. Until to the forth week, the ethanol concentration was increased to 55% for twelve weeks; Rats in normal control group were gavaged with physiological saline. At the end of 4, 8, 12 week, the rats in two groups were monitored with ultrasonography, and 3 rats in two groups were selected randomly for pathological observation. At last, the results of ultrasonography and pathology were analyzed comparatively. Results Ultrasonography and pathological examination results showed that alcoholic fatty liver model have been established successfully, and Ultrasonography can be used for monitoring gradual process of rats liver fatty lesions(from mild to heavy) in group model, and control rats without fatty lesions. The results of ultrasonography were consistent with the pathological examination results. Conclusion Ultrasonography can be better used for evaluating alcoholic fatty liver model in living rats.

Abstract:Objective To get histological data of laboratory animal for pathology inspection standard by comparing histological features of system main organ. Methods 30 of ordinary adult Macaca mulatta, 20 of Kunming mice, 20 of SD rat, 18 of Japanese big ear rabbit, 16 of Beagle, and 20 of Tupaia belangeri were selected for study. After intravenous anesthesia blood letting executed, trachea and lung were anatomised，fixed in 10% formalin and stained with HE, the organizational structure of the trachea and lungs were observed under microscope. The similarities and differences of lungs and trachea were compared among the different animal species. Results (1) Laboratory animals tracheal epithelial goblet cells have differences: The goblet cell of Macaca mulatta, Beagle, Japan big ear rabbit are more than those of SD rat ，Kunming mice and Tupaia belangeri . Epithelial mucus secretion type to neutral mucus is given priority to, Beagle goblet cells secrete mucus types have neutral mucus and acid mucus. (2) Laboratory animals submucosal gland bubble distribution difference: the submucosal gland bubble of Beagle are the most, Macaca mulatta, SD rat and Kunming mice, Tupaia belangeri too few quantity, Japanese big ear rabbit submucosal gland bubble at least. (3) Laboratory animals lung bronchial branch have difference: Beagle, Macaca mulatta, Japan big eared rabbit by lobar bronchus, segmental bronchus, bronchia, bronchi, terminal bronchi and respiratory bronchioles composition, SD rat and Kunming mice, Tupaia belangeri only by bronchi, terminal bronchi and respiratory bronchioles composition. (4) Laboratory animals bronchi organization structure have differences: Beagle, Macaca mulatta bronchi smooth muscle for complete circular ,there is no lack, SD rat , Kunming mice, Tupaia belangeri and Japan big eared rabbit bronchi smooth muscle thin or missing. Macaca mulatta, Tupaia belangeri and SD rat bronchi have a small amount of goblet cells,the rest of the laboratory animals were no goblet cells.(5) Laboratory animals Clara cell morphology are differences: Beagle Clara cell showed the cube,the rest of the animals is column. Conclusion There are some histological differences in trachea and lung of laboratory animals, which should be fully considered in drawing up pathology inspection standard, experimental research, or drug safety evaluation.

Abstract:Objective　To Screen suitable development stage of Mugilogobius chulae for acute toxicity test. Methods The prelarva, middle larva, postlarva, juvenile,young, adult of M.chulae were exposed to five concentrations of water-based drilling fluid and a control group and compare the acute toxicity of drilling fluid to M. chulae in different development stages. Results The 48h median lethal concentration(LC50) of the drilling fluid to M. chulae prelarva was 157 mg/L and to the middle larva was more than 500 mg/L. The 96h LC50 to the middle larva was 79 mg/L and to the postlarva was 625 mg/L.The 96h LC50 of the drilling fluid to juvenile, young and adult obviously exceeded 500 mg/L. That of the drilling fluid to Artemia nauplius was 105 mg/L.The sensitivity sequence of M.chulae in different development stages to drilling fluid was prelarva﹥middle larva﹥postlarva﹥juvenile, young and adult.Concolusion Prelarva of Mugilogobius chulae is suitable as laboratory organism for acute toxicity evaluation of marine pollutants

Abstract:Objective To set up an assay method of heart rate and noninvasive blood pressure determination in tree shrews，the data will be valuable reference values for developing tree shrews as an experimental animal. Methods 180 tree shrews were randomly divided in three groups，which included wild adult group, F1 generation artificial breeding adult and juvenile groups. Each group included 60 animals ( half female and male). The smart noninvasive blood pressure monitor mouse instrument（BP-98A）was used to measure HR(heart rate), SBP (systolic blood pressure), DBP (diastolic blood pressure) and MBP (mean arterial pressure). Results The heart rates of wild adult tree shrews, artificial breeding adult and juvenile tree shrews were 394.33?7.74BPM, 351.61?2.76BPM, 378.19?9.04BPM, respectively. There were be significant differences between wild adult and artificial breeding adult tree shrew groups (P<0.05). The SBP, DBP and MBP of artificial breeding adult group were lower than those of juvenile group, and there were great significant differences between two groups(P<0.01). There were no differences between wild adult and artificial breeding adult groups in SBP, DBP and MBP(P> 0.05). Conclusion We have established a technology for noninvasive blood pressure determination and obtained the reference values of heart rate and blood pressure from wild adult, artificial breeding adult and juvenile tree shrews. It will enrich the data of physiology in tree shrews, which provides scientific reference for studying on the animal experiment.

Abstract:Objective To prepare the polyclonal antibody against mouse HDAC2 and identify its reactivity and specificity. Methods An anti-peptide antibody has been developed by immunizing rabbits with a peptide synthesized according to the amino acid sequences of mouse HDAC2 protein. The peptide was coupled with KLH and injected into rabbits subcutaneously. The sera were collected and the antibody activity was evaluated with ELISA, Western blot and immunohistochemistry. Results The titer of the antibody in ELISA was estimated as high as 1:4000. Western blot assay of the brain extracts from the APP/PS1mice and HDAC2 peptide showed a positive band at 80kD when incubated with the antiserum. The immunohistochemistry stain show that the antiserum get the positive result with APP/PS1mice brain at the titer of 1:100, 1:200, 1:400, 1:800.Conclusion These antiserum have specific reactivity with HDAC2 in serum and tissue, and may possibly be applied in vivo and vitro in associated diseases.