Abstract:Objective To investigate the reproductive performance and compare pain threshold of ASIC3-/- and TRPV1-/- mice with C57BL/6 mice. Method: Twelve ASIC3-/- mice, twelve TRPV1-/- mice and twelve C57BL/6 mice were chosen and monogamously mated. There are 6 male and 6 female mice in each group, with age of 8-10 weeks. The pregnancy rate, the litter size, and the weaning rate of the ASIC3-/- mice and TRPV1-/- mice were analyzed. The pain threshold among the C57BL/6 mice, the ASIC3-/- mice and TRPV1-/- mice were compared. Result: The pregnancy rate, the litter size, and the weaning rate of the ASIC3-/- mice is slower than that of the TRPV1-/- mice. Comparing with C57BL/6 mice，the tactile threshold of ASIC3-/- mice was significantly higher (P<0.001) and the thermal threshold of TRPV1-/- mice was significantly higher (P<0.001). Conclusion The reproductive performance of the ASIC3-/- mice is weaker than that of the TRPV1-/- mice. The tactile sensitivity of the ASIC3-/- mice decreased. And the thermal sensitivity of the TRPV1-/- mice decreased.

Abstract:Abstracts: Objective To explore the chronic toxicity and carcinogenicity of pesticide pyraflufen-ethyl in rats. Methods According to National Standards of “Toxicological Methods of Pesticides for Registration” (GB15670-1995 of PRC), chronic toxicity tests and carcinogenicity tests were conducted. These samples were fixed in 10% neutral formalin firstly. Then the samples were embedded in paraffin after dehydration process. Afterward, the samples were sectioned and stained by hematoxylin-eosin(HE) staining. Finally, the sections were cemented by gum and observed in light microscope. Results Chronic toxicity tests revealed that the degeneration ratio of renal tubular epithelial cells in high dosage group was significantly increased comparing to control group (p≤0.01). Furthermore, the middle dosage group results showed there was a similar increasing in the proportion of tubulointerstitial nephritis (p≤0.05), however, there were no evident renal pathology differences between low dosage group and control group. In addition, other pathological changes in any dosage group were not remarkably different from control groups (p＞0.05). Statistical incidence variance of tumor, multiple primary neoplasms and tumor latency in different groups were not observed in carcinogenicity tests. Conclusion We found that the main target organ was kidney and sexual differences existed in chronic toxicity, in addition, the tumorigenesis of rats would not be elevated by Pyraflufen-ethyl.

Abstract:To investigate the protective effect of Apocynin on acute renal injury after ischemia/reperfusion (I/R). Methods Acute renal ischemia/reperfusion injury in male SD rats was made by cutting right kidney and clamping left renal artery.SD rats were randomly divided into groups(sham-operated group, I/R group, I/R+ Apocynin groups). Serum creatinine was assayed by biochemistry.Renal morphology changes were observed by light microscope. The amount of monocyte were observed by immunohistochemistry.MDA was assayed by thiobarbiturate. GPx was assayed by DTNB. IL-1 was assayed by Enzyme-linked immunoadsordent assay. Result Compared with sham-operated group, Scr levels were significant increased in I/R group（P<0.05）,kidneys displayd significant histology changes in I/R group（P<0.05）, the amount of monocyte was significant increased in I/R group（P<0.05）, MDA and IL-1 were significant increased in I/R group（P<0.05）， GPx was significant decreased in I/R group（P<0.05）. Compared with I/R group, Scr was decreased in I/R+Apocynin groups, the amount of monocyte was decreased in I/R+Apocynin groups（P<0.05）, MDA and IL-1 were decreased in I/R+Apocynin groups（P<0.05）. GPx was increased in I/R+ Apocynin groups（P<0.05）. Conclusions Apocynin protect kidney against ischemia/reperfusion and accelerate recovery after acute renal ischemia/reperfusion injury,which may associate with the anti-oxidation and anti-inflammation

Abstract:Esophageal variceal bleeding is critically ill patients, usually urgent medical conditions, and high mortality. How the prevention and control of bleeding, prevent further bleeding, reduce mortality, is still the focus and the difficulties in the present study. In this paper, esophageal varices establish animal models are reviewed.

Abstract:Oxidative stress and damage is closely related to skin functional decreased and diseases. Traditional biological evaluation of the skin antioxidant often use animal model through creative a model of local skin injury or disease. With the deepening understanding of the mechanism of skin oxidative damage and the development of in vitro culture techniques, using cultured skin cells or reconstructive skin model test system can be establish a new rapid testing method which can be used in the evaluation and screening of antioxidant effects, as well as mechanisms research and product development.

Abstract:【abstract】 Objective To explore the relationship of pulse pressure with renal function parameters in middle-aged patients with hypertension. Methods One hundred and Thirty-eight patients with hypertension were picked out of First Affiliated Hospital of Liaoning Medical examination in 2010-2012.Indexes such as blood pressure, age, serum creatinine was measured., creatinine clearance was estimated by the Cockcroft-Gault equations. And the relationship between pulse pressure and creatinine clearance was analyzed by Pearson test.Result Negative correlation was proved between pulse pressure and CCr.Conclusion Pulse pressure is one of the risk factors of renal dysfunction; creatinine clearance rate decreased with the increased pulse pressure in Middle-aged patients with hypertension

Abstract:Objective To investigate the management situation of Zhejiang Province of laboratory animals, and give suggestions of this science. Methods We designed some tables for this investigation, and investigated 61 units who have administrative licenses on the spot. Results Most of the units have perfect organization rules and regulations and SOP, but about 8.2% did not have contingency plan of laboratory animal, 24.6% never calibrate their autoclaves, 16.4% did not conduct sufficient verification of sterilization effects, and 4.9% units’ programs of processing of animal corpse and waste were not standard. Conclusion In order to improve the status of the subjects, and eventually promote the development of disciplines, and to adapt to the needs of the development of transformation, we should to further strengthen the management of autoclaves, and to formulate operable emergency plans.

Abstract:【Abstract】: Object: Research and experimental techniques of \"rabbit hemorrhagic shock\". Improve the successful rate of the experiment for a better effect of experimental teaching. Methods: 1.Student surveys and statistical analysis before and after the improvements.2. According to the experimental design of the experimental program.3. Animal experiments for improve experimental techniques and adjust the instrument is connected.4. Determine the technical path experiment and put into experimental teaching. Result: The much better teaching results. Conclusions:After the Improve of \"rabbit hemorrhagic shock\", The experimental results valid,operation difficulty is reduced,the successful rate significantly improved and better achieve the purpose of teaching.

Abstract:Objective Through comparing the drinking water, deionized water, pure water, double steamed water, purified water and sterilized water for injection and so on the various character water in vitro hemolysis test of hemolysis degree, we hope to provide the better positive control. Methods The hemocytolysis of various character water was observed by means of the routine test tube method in vivo (macroscopic observation methods) and modified hemolytic test method in vitro (spectrophotometry). Results Hemolytic crisis occur between the six character water, the hemolysis degree of sterilized water for injection is the largest, drinking water is minimum, the difference of several other character water is small, macroscopic observation method and spectrophotometry results are consistent. Conclusions In this test conditions, several character water to the rabbit red blood cells of hemolysis degree is sterilization water for injection > purified water > double steamed water > pure water >deionized water > drinking water, we recommended sterilized water for injection for hemolysis test positive control.

Abstract:Objective Investigate the application of in vitro fertilization (IVF) and embryo cryopreservation in C57-ras transgenic mice. Methods　 2-cell embryos were collected in two ways, first of which is by 4-week superovulated C57BL/6J female mice mated with C57-ras transgentic male mice, and the other is by IVF. The 2-cell embryos were cryopreserved after that. Then the cryopreserved 2-cell embryos were thawed and transplanted to caculate the ratio. Genetype of offsprings were analysed by PCR. Results The average recovery rate of cryopreserved normal 2-cell embryo was 67.78%?.08, and 21.31%?.09 were born after thawed 2-cell embryos transplanted. 23.08%?.14 of the offsprings were positive of human c-Ha-ras gene; The average recovery rate of cryopreserved IVF 2-cell was 63.33%?.03, and 15.56%?.04 were born after transplanted, and 33.33% of the offspring were positive . Conclusion The results suggested that there was no significant difference in cryopreservation of normal and IVF C57-ras 2-cell embryos. The research provided the cryopreservation method for C57-ras transgenic mouse model.

Abstract:【Abstract】 Objective To confirm whether the gene of PPARα imposes influences on the physiology, biochemistry index of mouse. The index of PPARα transgenic mouse is collected, analysed and the difference is obstained. The database content of transgenic mouse is enriched to provide the theoretical basis that can make the animal model apply in the preclinical drug evaluation. Methods The experiment animals were divided into 4 groups: PPARα male group（PPARα ♂）、PPARα female group (PPARα ♀)、C57BL/6 male group (C57BL/6 ♂)、C57BL/6 female group. (C57BL/6 ♀) . When the mice were at the age of 10 weeks, 14weeks, 18weeks, 22weeks, 34weeks, the blood routine , blood biochemistry of them were detected. At the age of 26 weeks，the pathological changes of heart, liver, kidney were detected .At the same time , the growth of them were observed. Results PPARα transgenic mice were compared with C57BL/6 mice ①The weight: When the animals were 6-23-week-old , there was no difference between the weight of PPARα transgenic mice and that of C57BL/6 mice（P＞0.05）. ②The blood routine : At the age of 10 weeks, the RBC、BAS%、LYM% of PPARα ♀ mice were obviously higher than those of C57BL/6 ♀ mice (P＜0.01) . At the age of 14 weeks, the WBC、LYM% of PPARα ♂ mice were obviously higher than those of C57BL/6 ♂ mice (P＜0.05). ③The blood biochemical index: At the age of 10 weeks, the BUN、LDL-C of PPARα ♂ mice were obviously higher than those of C57BL/6 ♂ mice (P＜0.05). At the age of 14 weeks, the AST、TG of PPARα ♂mice were obviously lower than those of C57BL/6 ♂ mice (P＜0.05). ④ The histopathology: Compared with C57BL/6 mouse,the heart of PPARα transgenic mice at the age of 26 weeks had no obvious change. The liver cell of them had transparent lesions , and the tubular epithelial cells had steatosis . Conclusion The weight, blood routine , blood biochemistry，pathology date of PPARα transgenic mice were systematically collected . The difference between PPARα transgenic mouse and C57BL/6 mouse was successfully analysed, and the referential date was provided for the application of the model.

Abstract:【Abstract】 Objective To explore the oxidative damage effect of methyl iodide on the liver of male rats. Methods Twenty-four male SD rats were randomly divided into 4 groups. Three experimental groups were given the methyl iodide with concentrations of 650mg/m3, 260mg/m3and130mg/m3 respectively for a week period of time. The duration of daily inhalation was 4 hours, The control group was given the same environmental conditions without exposure. At the end of the exposure，the level of superoxide dismutase (SOD) in serum and liver tissue and the activity of glutathione S transferase (GST) in serum were measured. Results Methyl iodide exposure can cause gradually increase of the SOD activity in rat serum, liver tissue and of GST activity in serum. There was significant difference, between experimental groups and control group. There was also significant difference among the low dose group, middle dose group and high dose group in the serum level of GST, Conclusions Methyl iodide exposure has the effect of oxidative damage on rat liver.

Abstract:Objective To investigated the therapeutical effects of Prescription No. 1 (PN-1) on Alzheimer’s disease by KinexTM Antibody Microarray which is a kind of high-throughput antybody microarray. Methods The Alzheimer’s disease mouse models are divided into treatment group and control group. There are 8 mouse in the treatment group and given once PN-1 every day by intragastric administration for 3 months，The 4 in the control group are given double distilled water every day by intragastric administration for 3 months too.The different expression protein between the two groups are screened by KinexTM Antibody Microarray , the molecules relevant Alzheimer’s disease is determined by Ingenuity Pathway Analysis（IPA）and critical review of literatures. Results Compared to control group, the proteins of the mouse in the treatment group which are associated with Alzheimer’s disease and have different expression between the two groups : p- tau（T522）、COX2、caspase6 and caspase9 which are decreased；bcl-xl and CaMK4 which are increased. Conclusions Prescription No. 1 (PN-1) can ameliorate the pathophysiological performance of Alzheimer’s disease mouse models by inhibiting the hyperphosphorylation of tau , inflammatory response,neuronal apoptosis and improving memory, is multitargeted，and may be an effective drug treatment for Alzheimer’s disease.

Abstract:Objective To observe parameters blood pressure, weight, organic weight and coefficient of the main organs on SD rats by alcohol interference. Methods Healthy Sprague-Dawley male rats were randomly divided into the blank group, the low dose alcohol group, the middle dose alcohol group, the high dose alcohol group. The low dose alcohol group, the middle dose alcohol group and the high dose alcohol group were respectively administered with 0.25mL/(100g bw.d), 0.5mL/(100g bw.d), 1.0mL/(100g bw.d) 52% alcohol through tube for 4, 8 and 12 consecutive weeks. During which time we recorded changes in blood pressure, weight, organic weight and coefficient of the main organs. Results Analysis of variance revealed no significant differences on blood pressure after 4 weeks, 8 weeks and 12 weeks of the SD rat induced by three doses of alcohol. After 4 weeks, there were significant difference in weight, heart weight, liver weight and kidney weight between the high dose alcohol group and the blank group (P<0.05); There were difference in coefficient of heart weight between the middle dose alcohol group and the blank group (P<0.05). After 12 weeks, there were difference in weight and coefficient of liver weight between the alcohol group and the blank group (P<0.05); There were difference in kidney weight between the high dose alcohol group and the blank group (P<0.05). Conclusions Biological parameters of SD rats with alcohol interference and normal SD rats are established which provide basic data for further study.

Abstract:Objective In order to research the development effect of icariin on the structure and function of mouse main organs. Methods 100 6-week-old female mice were divided into two groups randomly and fed with icariin and saline solution, at 0, 1st, 4th, 7th, 10th, 13th, 16th and 19th day, we measured the level of AST, ALT, ALP and LDH in serum by reagent-box, and the histological change of heart, liver, lung and kidney were observed. Result Icariin can condition the biochemical indicator of main organ obviously, increase the levels of AST and ALT in serum, decrease the levels of ALP and LDH in serum, but there is no change in organizational structure of main organs. Conclusion Icariin can affect the function of main organs but don’t change its structure.

Abstract:Objective The aim of this study is to revaluate the experimental animal model of diabetic retinopathy induced by streptozotocin (STZ) rats systematically. Medthods 75 Sprague- Dawley rats were divided into two groups randomly:30 for control and 45 for diabetic. Diabetes was induced by a single injection of 60 mg/kg STZ and monitored for 6 months. Body weight and blood glucose were determined dynamically. Three indexes of hemorheology were assayed. Retinal morphometry were determined by trypsin digest preparations. Expression of vascular endothelial growth factor (VEGF), pigment epithelium derived factor (PEDF) of retina were analysed by immunohistochemistry. Expression of occludin and intercellular adhesion molecule- 1(ICAM-1) of retina were measured by Western blotting. The activity of aldose reductase of retina was assayed by ultraviolet spectrophotometry. Results Compared with control group, the whole blood viscosity, plasma viscosity and erythrocyte aggregation index of STZ rats reduced, the basement membrane of retinal vessel thickened, numbers of capillaries degenerated with occlusion, retinal venular caliber dilated, the number of pericytes reduced obviously. The expression of retinal VEGF raised(IOD0.66?.28 vs control 0.25?.03), the expression of retinal PEDF declined(IOD0.07?.06 vs control0.19?.04)， occludin protein decreased to 70%, ICAM-1 protein increased by 2.58 times, the activity of aldose reductase increased by 3 times. Conclusion The diabetic retinopathy model in rat with STZ-induced diabetes occurs in multiple sites， and this animal model can be researched for diabetic retinopathy through multiple aspects and levels.

Abstract:Abstract : Objective To compare an optimized method of transverse aortic constriction (TAC) without ventilator and the traditional TAC by the survival rate of mice in one week. Methods Ninety C57/BL6 mice were randomly divided into sham-operated group (n=30), TAC without ventilator group (n=30) and TAC with ventilator group (n=30). The survival rates were recorded in 1 week and the cardiac function was evaluated by using ultrasonic instrument 4 weeks after the operation. Results The mice survival rate was higher in the TAC group without ventilator than in that with ventilator, and the new improved method resulted in the same cardiac hypertrophic model after 4 weeks. Conclusions The new optimized method of TAC without ventilator is better for improving survival rate and more convenient than the traditional TAC.

Abstract:【Abstract】 Objective To determine and analyze variation characteristics of heart rate (HR) and electrocardiogram (ECG) parameters in adult beagles within 24h. Methods The electrocardiogram was recorded by RM6240 multi-channel physiological signal acquisition decency via standard limb lead II in 12 adult beagle dogs which were under conscious state from 8:00 am to 8:00 am the next day. Results HR of beagle dogs showed a significant circadian rhythm with the fastest at 08:00 and the lowest at 12:00-14:00, HR at 08:00-09:00 was similar with that at 20:00-00:00. Corresponding with circadian rhythm of HR, P-R interval at 12:00 and S-T and Q-T interval at 11:00 was significantly increased, compared with that at 08:00 and 00:00 (p<0.05). Correlation analysis showed that there was remarkable correlation between HR and P-R, S-T and Q-T interval (P<0.01).Conclusion HR and part of ECG parameters in non-anesthetized adult beagle dogs showed a significant circadian rhythm, HR was slowly in the day, while P-R, S-T and Q-T interval prolonged correspondingly .

Abstract:Objective To establish an easy and efficient method of the isolation, purification and culture of Hamster primary hepatocytes. Methods Hamster Primary Hepatocytes were isolated using two-step collagenase perfusion in situ and purified by centrifugation with Percoll. The yields and viabilities were measured by standard trypan blue exclusion. Hepatocytes were identified using morphologic observation. Cell function was evaluated by Western blot method. Results Under the optical microscope, the obtained hepatocytes exhibited as irregular polygon with two or more circular nucleuses in each cell. The average viability of hepatocytes was 93.7% ± 2.1%. After incubated with adenosine, intracellular AMPK phosphorylation was significantly increased. Conclusion The method for isolating, purifying and culturing Hamster primary hepatocytes was successfully established.

Abstract:Objective BNIP3 mediate mitochondrial dynamic equilibrium in liver fat metabolism and glucose metabolism. Mitochondrial metabolism plays an important role in liver regeneration. Some studies have shown that EPO may promote liver regeneration, but the exact mechanism is remained to be elucidate. To study the effect of recombinant human erythropoietin (rhEPO) on liver function and liver regeneration after partial hepatectomy in rats, as well as the expression of BNIP3 and TNF-αmRNA in regenerating liver tissue. Methods 36 Wistar rats were randomly divided into the rhEPO group and blank group, and were submitted to 70% hepatectomy. The rhEPO group received 3000IU/kg of rhEPO throught portal vein injection and the blank control group received normal saline. Parameters of liver function and liver regeneration were assessed 1，3，5 days after 70% hepatectomy by means of automatic biochemical analyzer，immunohistochemical staining，ELISA assay and real-time PCR. Results 1 day after hepatectomy alanine aminotransferase (ALT) were significantly lower in rhEPO group than the blank group (P <0.05). 1,5 day after hepatectomy Ki-67 labeling rate in rhEPO group were significantly higher than the blank group (P <0.05). 1day after liver resection serum TNF-α in rhEPO group were significantly higher than the blank group (P <0.05), 1,3d ays after hepatectomy serum IL-6 in rhEPO group were significantly higher than the blank group(P <0.05). 1,3days after liver resection, BNIP3 and TNF-α mRNA in rhEPO group were significantly higher than the blank group (P <0.05). Conclusions Portal vein injection of rhEPO can protect liver function and promote liver regeneration, the mechanism may be related to BNIP3 and TNF-α.