Animal experiment is one of the most important method in life science research. In this article,factors affecting the results of animal experiment were summarized,which include laboratory animal,environment,nutrition,chemicals,isolation and feeding density,transportation,grasping and fixing,microbiology,society and people etc.

Allogeneic stem cell transplantation is the only treatment which may cure multiple myeloma (MM).The high transplantation related mortality (TRM) limits the wide clinical application of myeloablative allogeneic stem cell transplantation.Non-myeloablative allogeneic stem cell transplantation can reduce the TRM,but the risk of relapse and progress of the disease may get increased. Autononmyeloablative allogeneic stem cell tandem transplantation can reduce both the TRM and the relapse and progress of the disease. At present,there exist many new methods which may improve the effect of allogeneic stem cell transplantation for MM in clinical.

Comparative medicine is a synthetical subject,to provide protection and improve healthy medicare of human being by studying all kinds of human diseases using laboratory animals.Comparative medicine is widely involved in training medical talents,elucidating disease mechanism,improving medicare level and benefiting human healthy services.Therefore,comparative medicine research is an important foundation of medicine development,and indispensable supporting condition and organic compartment in all-round development of "medication,education and research" in modern comprehensive hospital.

[Abstract ] Objective Evidence from previous studies indicated that over-activation of C3a/C3aR axis existed in the renal tubular epithelial cells of patients with renal diseases including diabetic nephropathy .However , the pathological significance of over-ac-tivating C3a/C3aR axis still remains to be elucidated .In this study, we constructed a renal tubular epithelial cell line over-expressing C3a in a secretory manner in order to provide a cell model to investigate the pathological significance of over -activating C3a/C3aR axis under various pathological scenes . Methods We designed a synthesized C3a secretory expression unit and cloned it into the multi-clonal site of lentivirus expression vector pLenti 6.3-MCS-IRES2-EGFP.After identification by sequencing , recombinant lentivirus was packaged by using pLenti 6.3-C3a-IRES2-EGFP and packaging plas-mid in 293T cells.Then, the recombinant lentivirus was used to in-fect HK2, a cell line of human renal tubular epithelial cells .After screening in medium with blasticidin , blasticidin resistant cell clones were obtained .Real-time PCR and ELISA method were applied to analyze the expression and secretion of stable transfected cells cloned C3a and identify renal tubular epithelial cell lines with stable over-activating C3a. Results ①C3a secretory expression unit was suc-cessfully synthesized and correctly cloned into the multi-clonal site of pLenti6.3-IRES2-EGFP; ②C3a secrectory expression recombi-nant lentivirus LV-C3a was successfully packaged with a high titer of 5 ×108/mL;③HK2 Cell clones resistant for blasticidin were ob-tained;according to the analysis of Real-time PCR and ELISA, the C3a mRNA level in HK2-C3a cell lines was significantly higher than that of HK2 cells(1.0 ±0.5 vs 1321.0 ±18.0, P<0.01) and the secreted C3a level increased significantly ([0.3 ±0.2]ng/mL vs [249.0 ±37.0] ng/mL, P<0.01). Conclusion The present study successfully constructed C 3a secretory expression vector pLenti6.3-C3a-IRES2-EGFP and C3a over-expression renal tubular epithelial cell line HK 2-C3a, which is very useful in further study of the function and significance of C 3a/C3aR axis not only in renal tubular epithelial cells but also in other cell types .

Objective Increased renal mast cells have been found in significant correlation with clinicopathological features of diabetic nephropathy ( DN) .However, the exact pathological significance of mast cells still remained to be elucidated.As one of the most abundant serine proteases, tryptase is specifically expressed by mast cells.The study was to observe the expression of tryptase in the urine of patients with diabetic nephropathy and realize the pathological significance of urinary tryptase and renal mast cells in the development of diabetic nephropathy. Methods Urine samples from 103 patients with diabetic nephropathy who were hospitalized at the clinical unit of the nephrology centre of Jingling Hospital from January 2012 to June 2013 were collected.According to concentra-tion of urinary albumin excretion rate, the patients were conducted to early-stage DN group (microalbuminuria,n=10), middle-stage DN group (proteinuria stage, n=31) and end-stage DN group (renal insufficiency stage,n=62).Meanwhile, urine samples from 20 normal persons were taken as the normal control group.Tryptase level in each urine sample was measured by using an ELISA kit.The average tryptase levels were compared among different groups and the correlation between urinary tryptase level and clinical indices was analyzed. Results ①In most cases, tryptase was not detected in the urine samples from normal persons(0.7 ±1.4 ng/mg creati-nine).However, the urinary trypatse level increased significantly with the development of diabetic nephropathy (11.6 ±10.5 ng/mg creatinine for early stage, 29.7 ±19.2 ng/mg creatinine for middle stage, and 44.6 ±43.4 ng/mg creatinine for late stage group).②The urinary tryptase level was found in significant correlation with ser-um creatinine (r=0.325, P<0.01), serum cystatin C (r=0.391, P<0.01), serum urea nitrogen (r=0.27, P<0.01), 24 hour uri-nary protein (r=0.389, P<0.01), urine C3 (r=0.276, P<0.05), urine retinol-binding protein (r=0.365, P<0.01), urine lysozyme (r=0.461, P<0.01), urine N-acetyl-β-glucosaminidase level (r=0.568, P<0.01), urinary kidney injury molecule-1 (r=0.434, P<0.05) and interleukin-18 level (r=0.375, P<0.05).No significant correlation was found between urinary tryptase level and age, gender, fasting blood sugar, postprandial blood sugar, HbA1c, triglyceride, high density lipoprotein or low density lipo-protein. Conclusion Mast cells play an important role in the development of diabetic nephropathy and might be a novel and effective target for the treatment of DN.

Objective To analyze the clinical characteristics ,diagnosis and treatment of acute leukemia complicating deep ve‐nous thrombosis(DVT) to deepen the cognition on this complication .Methods The clinical data of consecutive patients with acute leukemia were performed the retrospective analysis .The occurrence situation of deep venous thrombosis was investigated and the a‐broad related literatures were reviewed .Results A total of 116 cases of acute leukemia in our department from July 2011 to March 2014 were treated ,in which 85 cases were acute myeloid leukemia and 31 cases were acute lymphoblastic leukemia;5 cases devel‐oped DVT with the proportion of 4 .31% (5/116) .Of these cases ,3 cases were acute promyelocytic leukemia and 2 cases were acute lymphoblastic leukemia .Conclusion The occurrence rate of DVT in the patients with acute promyelocytic leukemia is relatively higher ,if the patients have the corresponding symptom ,timely diagnosis and treatment should be conducted .

Objective To explore the source of endothelial cells in tumor vessels by A 549 tumor model with GFP nude mouse.Methods To establish the A 549 lung cancer models with GFP nude mice, expression of CD 31 was determined by immunofluorescence to label tumor vessels; to observe and take a picture of the tumor frozen section by confocal microscopy and invert microscope;expression of GFP in tumor vessels was determined by immunohistochemistry. Result The results of immunofluorescence showed:Tumor interstitial vascular endothelial cells or endothelial cells clusters and micro-vascular lumen size and shape are clearly visible by immunofluorescence, and part of vessels with no obvious lumen or irregular lumen.We can see green fluorescent in tumor cells of tumor tissue and endothelial cells which form of tumor vessels.The results of immunohistochemistry showed: expression of GFP was determined in cytoplasm of tumor stromal cells and endothelial cells in tumor vessels.Conclusion The endothelial cells which formed tumor neovessels that derived from GFP nude mice partly and the other part derived from tumor cells.

Objective There are a few reports on rats with spontaneous congenital cataract in China .The purpose of this study was to investigate the morphological changes of lens in rats with spontaneous congenital cataract . Methods 24 d, 1-year rats with cataract and microphthalmos cataract and normal rats (n=5) were selected as research objects .Their lens were observed by a slit lamp microscope and taken photos in front of them , followed by examination through light micrograph and transmission electron micros-copy. Results Rats with microphthalmos cataract showed narrowed palpebral fissure and broaden nucleus while rats with cataract showed normal palpebral fissure and narrowed nucleus .As for 24 d,1-year rats with microphthalmos cataract , the fibers of their lens showed derangement and vacuole-like degeneration by light microscope , in addition, the abnormal connection between fiber cells were observed by electron microscopy .As for 1-year normal rats , the fibers were in consistent structure and regular arrangement without cell ingredient . Conclusion The appearance and morphological changes of the lens in rats with spontaneous congenital cataracts are in consistence with the pathological changes of cataracts , which is appli-cable in further research on the pathogenesis of cataract .

Objective To measure the range of routine blood counts and blood biochemical parameters ,to analyze hemorheology of the rats with congenital cataract .Methods Blood samples were taken from 90 rats with congenital cataract weight about 185 ~211 g.Routine blood analysis was performed and blood biochemical and hemorheology paramenters were determind using an automatic blood biochemical and hemorheology analyzer .Results There were no significant difference ( P >0.05 ) between the cataract rats and the normal rats in Blood test results; but there were significant difference between the microphthalmos cataract rats and the normal same -sex rats ( P <0.01 or P <0.05 ) . The biochemical results is the cataract rats and the normal rats were different significantly in ALB group ( P <0.01 or P<0.05), and the female microphthalmos cataract rats compared with the control rats had significant difference in Ure (P<0.01) , the female cataract rats ompared with the normal rats were very significant difference in Cr group ( P <0.05 or P <0.01).The erythrocyte counts of the male cataract rats and male microphthalmos cataract rats were significantly lower than that in the female ones, respectively(P <0.05, P <0.01).The platelet counts of the male cataract rats and the male microphthalmos cataract rats were significantly higher than that in the female ones , respectively(P <0.01), and the creatinine of the male cataract rats and the male microphthalmos cataract rats were significantly lower then that in the female ones, respectively(P <0.01).There were no significant difference in every group on hemorheology .Conclusions There were significant differences in some blood indexes between the congenital cataract rats and the normal rats .These data may become useful reference for biomedical researcher in this field .

Objective:To design the mode of supervision, forwarn and communication system in barrier enviroment of laboratory animal. Methods and Results:Distribution inspect system of industrial computer center was used for live monitoring,displaying,reserving, printing temperature humidity,pressure,difference and ammonia density,according to necessity.The upper and low limit of parameter was set up according to national standard of laboratory animal,if parameter surpass the limit,it will warn the people on duty in multiple spots.Working and animal illumination were established respectively on the basis of illuminace light period of standard,the switch of working illumination was controlled by the people,but the later was controlled, automatically and slowly in order to disinterfere with physiology activity of animal.The mode of moniter add video together with multiple pickup camera in cable television were used,which binded wirh the intellectual industral control group.The communication system was designed using miniature program control changer and terminal telephone, which joined the unit inner telephone to communicate with outside. The double and stereophonic was used to install background music in this enviroment,which was timely played with suitable volume. Conclusion:This system provided the method for the administration of laboratory animal standardization?mathematics and informatics.