Abstract:Objective The functional status of the host immune system plays an important role in viral infection．The aim of this work was to study the differences among four immunodeficiency strains of mice and two wide-type mouse strains after infection of influenza A ( H1N1) ( A /California /7 /2009 ( CA7) ) virus ． Methods The six inbred laboratory strains of mice were anesthetized by inhalation of ether and inoculated intranasally with 0. 05 mL of infectious virus H1N1 ( CA7) suspension． The body weight loss kinetics，survival rates and pathological changes in the lung tissue after infection were examined． Results BALB / c mice and four immunodeficiency strains of mice showed high susceptibility in the viral infection compared with C57BL /6 mice． Half of C57BL /6 mice survived，while the mortality rates of the remaining five strains were 100% ． C57BL /6 mice exhibited diffuse interstitial pneumonia in the initial stages of infection，and the lesions were gradually restricted to the late stage． BALB / c mice and four immunodeficiency strains of mice showed severe diffuse interstitial pneumonia，bronchial epithelial degeneration and necrosis，and there were significantly less inflammatory cells than that in the C57BL /6 mice． Conclusions Innate immunity and adaptive immunity play a major role at the early and late stages of the infection of influenza A ( H1N1) virus． The state of the host immune system affects the infection and prognosis of influenza A ( H1N1) virus infection．

Abstract:Objective To generate a transgenic mouse line expressing human APP695 K595N / M596L ( Swedish mutation) and establish a transgenic Alzheimer disease model． Methods The transgenic plasmid was constructed by inserting the mutated APP695 K595N / M596L gene into the downstream of mouse prion protein promoter． The transgenic mice were produced by microinjection and the genotype was detected by PCR． The gene expression levels were determined by Western blotting． The senile plaques were detected by thioflavin-S staining and visualized directly by fluorescence microscopy． The behavioral changes was examined by Morris water maze test． Results Transgenic C57BL /6J mice were generated with the expression of the APP695 K595N / M596L in the brain tissue． The transgenic mice showed significant learning and memory impairments in the Morris water maze at 5 months of age and the extent of the impairments was developed at 7，9 and 11 months of age，comparing with that of age-matched wild type mice ( P ＜ 0. 05) ． Senile plaques were visualized in the CA1 area of hippocampus at 9 and increased at 12 months of age． Conclusions The transgenic mice show a progressive deficits of learning and memory and a progressive formation of senile plaques in the hippocampus，suggesting that this APP transgenic mouse is an useful animal model of Alzheimer disease．

Abstract:Objective To investigate the effects of SHP-2 D61G / + mutations on leukocyte infiltration，cytokines secretion and multi-organ impairment． Methods The SHP-2 D61G / + gain-of-function mutant gene knock-in model mice and wild-type 57BL /6 mice ( WT) were used in this study． Serum level of IL-2 and TNF-α and the cytokines concentration in the supernatant of cultured WBC were quantified by enzyme-linked immunosorbent assay ( ELISA) ． Leukocyte infiltration in the lung，spleen and heart tissues was observed by histopathology，and serum levels of alanine aminotransferase ( ALT) and cardiac troponin I ( cTnI) were studied by radioimmunoassay． Results Compared with the WT control，leukocyte infiltration in the lung and spleen tissues were significantly increased and myocardial hypertrophy was observed in the SHP- 2 D61G / + mutant mice． The serum levels of IL-2 and TNF-α were significantly increased in SHP-2 D61G / + mutant mice ( P ＜0. 01，compared with the WT control) ，and so were the cytokines concentrations in the supernatant of cultured WBC ( P ＜0. 01，compared with the WT control) ． The serum levels of ALT and cTnI were significantly higher in the SHP-2 D61G / + mutant mice than that of the WT controls ( P ＜ 0. 01 ) ． Conclusions Gain-of-function mutations in SHP-2 tyrosine phosphatase ( SHP-2 D61G / + ) greatly induce leukocyte infiltration in tissues，IL-2 and TNF-α secretion and multi-organ impairment，which may lead to multiple organ failure．

Abstract:Objective To investigate the effects of a Chinese traditional medicine，Tong-guang-san ( TGS) ，on airway hyperresponsiveness in asthmatic mouse models． Methods Thirty five BALB / c mice at 6 weeks of age were randomized into 3 groups． The mice of model group and experimental group were sensitized with ovalbumin ( OVA) by intraperitoneal injection on day 0，7 and 14，then challenged with OVA by intranasal administration on day 28，29，30 for asthmatic models． From on day 15，the mice of the experimental group were administered TGS ( 0. 72 mL，about 0. 04 g crude drug ) via intragastric gavage once daily for 14 days． The mice of control group received saline． Bronchial hyperresponsiveness ( BHR) was examined 48 hours after the final challenge on day 32 with a whole body plethysmography ( WBP) system． Bronchoalveolar lavage fluid ( BALF) and histological changes in the airways were examined on day 32. The data were analyzed statistically with one-way analysis of variance ( ANOVA) using SPSS package ( version 13. 0 ) ．Results Airway resistance of the mice in the experimental group was significantly decreased compared with that of the asthma groups ( P ＜ 0. 05) ． In asthma group，the Eos percentage in BALF were 70% but those cells were scarcely seen in the normal group ( 0. 08% ) ． Eos in BALF in the experimental group were decreased markly compared with that in the asthma group ( P ＜ 0. 05 ) ． The histological examination of the lungs from the asthmatic animals revealed characteristic inflammatory cell infiltration in the peribronchial regions and around the submucosal blood vessels while those of experiment group displayed less inflammation and improved mucous edema and epithelial lesions of the bronchi and bronchioles．Conclusions TGS can significantly reduce airway hyperresponsiveness of the asthmatic mice．

Abstract:Objective To determine the optimal dosage of Folium Artemisiae argyi in air disinfection of animal facility and respiratory irritation to animals． Methods 1. Three different dosages of Folium Artemisiae argyi，1 g /m3，2 g / m3 and 4 g /m3，corresponding to low，middle and high dosage groups，were used in air fumigation disinfection of empty animal facility，respectively． Colony forming units ( CFU) of bacteria was examined before disinfection and 12 hours after disinfection，once a week，for ten weeks． The eligible standard of CFU was ≤30，The eligible rate was calculated in these groups after 10 times of disinfection． 2. Forty SD rats were grouped into 4 groups，the control and 1 g /m3，2 g /m3，4 g /m3 dose groups，each group has a disconnector． Fumigation disinfection was done with 1 g /m3，2 g /m3 and 4 g /m3 Folium Artemisiae argyi in different groups． The lungs of rats were taken out to observe the histopathological injury after 12 hours．Results 1. CFU assay of bacteria was not eligible in all groups before disinfection，but it was eligible in every experimental group after 10 hours disinfection． There were significant differences between the low dosage group and middle，high dosage groups，and no significant difference between middle dosage group and high dosage group． The eligible rate was 80% in low dosage group and 100% in middle and high dosage groups． 2. Pathologicl injury of lungs was observed in all experimental groups． Conclusions The optimal dosage of Folium Artemisiae argyi in air disinfection of animal facility is 2g /m3，meanwhile，it may irritate the respiratory system of experimental animals．

Abstract:Objective To observe the intestinal pathological changes of dead rhesus monkeys bred under captive conditions，and to explore the rule of distribution of characteristic pathological changes． Methods One hundred and fiftyfive naturally died monkeys in the age of 2 to 20 years，bred in institutes in Yunnan province，were autopsied from 1998 to 2008. The monkeys were divided into young，adult and old groups according to their age． The intestinal changes were investigated by routine histopathological examination and analysis by chi-square test． Results Of the 155 monkeys，58 animals had intestinal lesions ( 37. 42% ) ． The main lesions included chronic enteritis，acute enteritis，bleeding and erosion，ulcers，perforation，parasite infections and others． All kinds of intestinal lesions occured in the 3 age groups． Among these lesions the incidence rate of acute enteritis is the highest ( 20. 00% ) and increased with increasing ages． The chi-square test showed that the distribution of main intestinal lesions had no statistically significant difference among the 3 age groups． Conclusions The incidence rate of intestinal diseases in the bred rhesus macaques is high． The intestinal changes are not correlated with the ages of the captive rhesus macaques． Severe acute eneritis is the main cause of death of the captive monkeys． We should pay enough attention to the intestinal lesions，especially acute enteritis，when raise and study rhesus monkeys． The results of our test provide useful information in quality control of captive monkeys and relevant animal studies．

Abstract:Aim To explore the effects of substance P ( SP) and calcitonin gene-related peptide ( CGRP) on the occurrence and development of inflammatory bowel disease ( IBD) ． Methods The IBD rat model was generated by chemical induction with trinitrobenzene sulfonic acid ( TNBS) ． The expression of SP and CGRP in the colon tissues was determined by real-time RT-PCR． Results With the development of pathological changes ( on days 3，7) ，the expression level of SP and CGRP in the colon was significantly increased，and then followed by a decline and gradual return to normal levels ( on days 21，28) ． Generally speaking，the expression of both SP and CGRP was positively related to the occurrence and development of IBD． Conclusions The results of this study revealed that the neurotransmitters，SP and CGRP，play an important role in signal transduction in the inflammation and repair of inflammatory bowel disease

Abstract:Objective To compare two types of anastomosis in a rat model of intimal hyperplasia of autologous vein graft in rats． Methods Twenty SD rats were randomly divided into two groups． The external jugular veins were anastomosized with the common carotid artery of the same side with interrupted suture and twice continuous suture，respectively． Tissue samples were harvested at 4 weeks after the operation． The vein graft patency was observed and the thickness and area index of the neointima and media were analyzed by histopathology with HE staining． Results Compared with the interrupted suture group，the twice continuous suture showed less time consuming，less bleeding，but also less graft patency． The intimal hyperplasia in the vein grafts occurred after the two anastomosis methods had no significant difference． Conclusions The twice continuous suture has advantages of less time consuming and less bleeding，but it requires more skill． There is no significant difference in the degree of intimal hyperplasia of autologous vein grafts in rats by the two anastomosis methods．

Abstract:Objective To study the mechanisms responsible for the changes in alveolar fluid clearence in rats with malnutrition． Methods Rats were fasted but allowed access to water for 120 hours． Alveolar fluid clearance ( AFC) ，lung water content ( TLW) and extravascular lung water ( EVLW) of the rats were calculated in the rats with malnutrition．Isotonic 5% album in solutions with amiloride，ouabain and terbutalin were instilled into the distal airways in the malnutrition and control rat lungs，and the AFC was examined． Results As compared with the control rats ( 18. 5 ±2. 21% ) ，AFC in the rats with malnutrition for 48 h was not decreased ( 19. 7 ± 3. 19% ) ． AFC in the rats under conditions of malnutrition for 120 h was decreased ( 9. 50 ± 2. 19% ) ． Amiloride and ouabain could reduce the AFC in both malnutrition and control rats． There was no significant difference in the magnitude of the terbutalin-stimilated AFC in rats under conditions of malnutrition compared with that in control rats． Conclusion Malnutrition primarily impairs amiloridesensitive and amiloride-insensitive alveolar fluid clearance，leading to pulmonary edema．

Abstract:Objective To observe the characteristics of coronary artery ligation and coronary embolism for preparation of acute myocardial infarction ( AMI) models in canine． Methods The left main anterior descending coronary artery was obstructed by either open thoracotomy ligation or closed-chest coronary embolism． The electrocardiogram changes were recorded before and after blocking coronary blood flow． At 72 hours after the acute myocardium infarction model preparation，the dogs were sacrificed and specimens of the myocardium were taken for histopathological examination．Results Both electrocardiogram and histopathology confirmed that both methods could establish AMI models successfully．Compared with open-chest coronary artery ligation，the closed-chest coronary embolism had a higher survival rate．Conclusion Closed-chest coronary artery embolism deserves recommendation due to the advantages such as safety and convenience in the preparation of dog models of acute myocardial infarction．

Abstract:Objective To determine the complete sequences of the mitochondrial DNA control region in Macaca mulatta from Yunnan province． The complete sequences were analyzed for phylogenetic tree and nucleotide composition．Methods The complete sequences was amplified by polymerase chain reaction ( PCR) ． The nucleotide composition was analyzed by comparison with the known control region sequence ( AY612638) downloaded from GenBank database． The homology，the rate of transition /transversion and phylogenetic tree constructed by neighbor-joining ( NJ) and minimumevolution ( ME) methods were also analyzed． Results The length of the mitochondrial DNA control region complete sequences were 1084 to 1089 bp，the average contents of A，T，G and C were 29. 9% ，26. 9% ，12. 3% and 30. 9% ，respectively． The content of A + T ( 56. 8% ) was higher than G + C ( 43. 2% ) in the complete sequence． The homology of these sequences was 91. 5% ～ 99. 5% ． The nucleotide mutational rate was 4. 5% on average，including transition，transversion，insertion and loss． The rate of transition /transversion was 26. 1 on average． The phylogenetic analysis based on mitochondrial DNA control region showed that the Macaca mulatta of Yunan was divided into two sister groups geographically． Conclusions The complete sequences of the mitochondrial DNA control region of Macaca mulatta has been determined． This study may be of importance for studies on evolution and function of the mitochondrial DNA control region of Macaca mulatta．

Abstract:Objective To observe the regulatory effects of Smilax glabra on blood pressure and vasoactive substances in renovascular hypertensive rats． Method Seventy healthy adult male rats were used to establish the model of renovascular hypertension by two-kidney two-clip procedure． Model rats were divided into model control group，high ( Smilax glabra 6 g / kg) ，moderate ( Smilax glabra 3 g / kg) ，low ( Smilax glabra 1. 5 g / kg) dose groups and the positive control group ( captopril 5 mg / kg) according to systolic blood pressure in the renal hypertensive rats． Rats of the drug groups were administrated the corresponding drugs by once daily for 4 weeks，and measured blood pressure once a week．After the last administration，the blood ANP，NO，ET，CGRP and Ang II of the rats were measured． Result Smilax glabra significantly reduced SBP，DBP and MBP in the renal hypertensive rats ( P ＜ 0. 05，P ＜ 0. 01 ) ，significantly reduced the ANP，ET levels ( P ＜ 0. 05，P ＜ 0. 01) ，and increased the NO level ( P ＜ 0. 05) ． Conclusion Smilax glabra has a blood pressure lowering effect and regulates the blood pressure by reduced ANP，ET，and increased NO levels．

Abstract:Objective Two non-radioactive methods for determining glomerular filtration rate ( GFR) in conscious rat using FITC-labeled inulin ( FITC-inulin) and micro-osmotic pumps were evaluated． Methods FITC-inulin ( 24% ) was dissolved in 0． 9% NaCl and the concentration decreased to 8% after 24 h of dialysis． Two micro-osmotic pumps filled with 200 μL of 8% FITC-inulin were inserted into the peritoneal cavity of rats． After their complete recovery from anesthesia，the rats were housed individually in metabolic cages． Urine and the residual fluorescence remaining on the cages were collection over 24 h on day 7 after micro-osmotic pump implantation． Blood sample was collected through the saphenous vein at the end of 24-h urine collection． Only blood was sampled using the same method in another group． GFR was evaluated on day 7 after micro-osmotic pump implantation using two methods expressed in microliters per minute，microliters per minute per kilogram body weight and per gram kidney weight． Results Based on the approach measuring urinary inulin clearance with urine collection and without urine collection the estimated GFR was ( 2. 31 ± 0. 33) microliters per minute and ( 2. 53 ± 0. 33) microliters per minute ( P = 0. 564) ，respectively． These values of GFR in conscious rats were only ca． 70% of that obtained in anesthetized rats determined in other previous studies． It was revealed that anesthesia may significantly influence GFR． Conclusions The results of the present study demonstrate the feasibility of the osmotic micropump approach to monitor GFR in conscious rat using FITC-inulin． Especially，the method without collecting urine is more convenient．

Abstract:Objective To evaluate the feasibility of endotracheal intubation route in preparation of H5N1 virus infection in rhesus macaques and to obtain a more sensitive animal model． Methods The rhesus macaques were infected with H5N1 avian influenza virus by endotracheal intubation． Clinical signs were assessed daily，and throat swab，nasal wash，major organs and blood were collected for detection of virus load，isolation and blood routine analysis． Results After infection the animals exhibited anorexia，lethargy and transient fever，and the total number of circulating leukocytes and lymphocytes was decreased． Virus isolation，RT-PCR assay of the throat swabs，nasal washs materials showed that the intestine，lung，brain，spleen，kidney，heart and liver were the target of the virus replication and the virus could replicate in organs outside the lung． In addition，virus was also detected in the blood 3 days after infection． Conclusions Rhesus macaques can be infected with H5N1 virus by endotracheal intubation，and it is proved to be a good method to obtain a more sensitive animal model of H5N1 virus infection． The experimental results will lay the foundation for further studies of pathogenetic mechanism of H5N1 infection．

Abstract:Objective To establish a miniature pig model of abdominal wall biological zipper，to study its biological characteristics，and to provide a useful and convenient tool for research and teaching． Methods A selfconstructed special biological zipper was fixed on the abdominal wall of miniature pigs by surgery，and to establish an animal model of abdominal wall zipper． The changes of physiological and biochemical indexes of blood and urine of the miniature pig models were measured at 0，1，3，7，14，21，28，35，49，56 days after surgery and the data were analyzed． The blood physiological and biochemical parameters were RBC，Hb，PLT，total proteins ( TP) ，albumin ( ALB) ，total cholesterol ( TC) ，triglyceride ( TG) ，sodium ( Na + ) ，potassium ( K + ) ，and chlorine ( Cl － ) ． The urine parameters included glucose ( Glu) ，proteins ( Pro) ，bilirubin ( BIL) ，urobilinogen ( URO) ，specific gravity ( SG) ，pH，leukocytes( WBC) ，erythrocytes ( RBC) ，and nitrite ( Nit) ． Results The general condition，daily behavior，physiological and biochemical parameters of blood and urine of the miniature pig models showed no significant changes during the 7 － 49 day period after modeling，compared with those in the control pigs． Conclusions A miniature pig model of abdominal wall biological zipper has been successfully established in this study，showing stable physiological and biochemical functions during the 7 － 49 day period after modeling． This model is suitable for wide application in relevant research and teaching works．

Abstract:Objective To identity the prevalence of Helicobacter spp． infection in laboratory mice and laboratory rats in the area around Shanghai，and provide reference and basis for the establishment of Chinese laboratory animals grades and monitoring standards． Methods A total of 352 mice ( 101 clean grade mice，251 SPF grade mice) and 101 rat ( 69 clean grade rats，32 SPF grade rats) obtained from the area around Shanghai were detected by polymerase chain reaction ( PCR) ． 88 mice ( clean grade mice，62 SPF grade mice) and 165 rats ( 84 clean grade rats，81 SPF grade rats) were detected by enzyme linked immunosorbent assay ( ELISA) ． Next，the positive rate of the two methods in 88 mice and 101 rats were compared． Results PCR detecting percentage in mice and rats were as follows: the average positive rate of mice was 35. 8% ( 126 /352) ，among them，the positive rates of clean grade rats and SPF grade mice were 51. 5% ( 52 /101) and 29. 5% ( 74 /251) ，respectively． The average positive rates of rats were 70. 3% ( 71 /101) ，among them，the positive rate of clean grade and SPF grade were 69. 6% ( 48 /69 ) and 71. 9% ( 23 /32 ) ，respectively． ELISA detecting percentage in mice and rats as follows: the average positive rate of mice were 15. 9% ( 14 /88) ，among them，the positive rates of clean grade and SPF grade mice were 19. 2% ( 5 /26) and 14. 5% ( 9 /62) ，respectively． The average positive rate of rats was 52. 7% ( 87 /165) ，among them，the positive rates of clean grade and SPF grade rats were 53. 6% ( 45 /84) and 51. 9% ( 42 /81) ，respectively． The positive rates of 88 mice tested by PCR were 72. 7% ( 64 /88 ) and by ELISA 15. 9% ( 14 /88 ) ． The positive rates of 101 rats detected by PCR were 70. 3% ( 71 /101 ) and by ELISA 49. 5% ( 50 /101 ) ． Conclusions Helicobacter spp． infection exists in laboratory mice and laboratory rats in the area around Shanghai． The PCR assay of feces is more sensitive than the examination of the serum antibody against Helicobacter spp．

Abstract:The anatomy and physiology， glucose-lipid metabolism， blood biochemical parameters and the mechanism of diabetes in minipigs is very similar to that of humans． Its price is also relatively modest． So minipigs are excellent animal models of induced diabetes． This article is a review on the progress in research on minipig models of diabetes in recent years．

Abstract:Objective To establish an evaluation index system for the laboratory animal management in provincial medical universities． Methods Thirty seven experts were selected according to the research topics，and the Delphi method was used for questionnaire survey and statistical analysis． Results There was a 70. 27% response rate of the first investigation and a 76. 92% response rate of the second investigation． The authority coefficient was 0. 855 in the first investigation． The harmonious coefficient was 0. 486 of the second investigation． Conclusions The evaluation index system for the laboratory animal management in provincial medical universities is a reliable and useful system．