Abstract:The use of animals in research will be necessary for scientific advances in the basic and biomedical sciences for the foreseeable future． As we learn more about the ability of animals to experience pain，suffering，and distress，and particularly for mammals，it becomes the responsibility of scientists，institutions，animal caregivers，and veterinarians to seek ways to improve the lives of research animals and refine their care and use． Refinement is one of the three R’s emphasized by Russell and Burch，and refers to modification of procedures to minimise the potential for pain，suffering and distress． It may also refer to procedures used to enhance animal comfort． This paper summarizes considerations for refinements in research animal．

Abstract: Objective Methods 28 male Mongolian gerbils were divided into 4 groups，7 per group． Set a negative control group，three CP groups ( low-dose group，medium-dose group and high-dose group) ． Cyclophosphamide was given to the CP groups by intraperitoneal injection for 5 days． 30 days after the giving drugs，we anaesthetized the gerbils，opened their abdomen and took out their two testicles． Then we made smears with the sperm suspension，and air dried them，and then fixed them with methanol for 10 minutes，and then air dry them again． After the above steps，we stained them with 2% eosin for 30 minutes，then we rinsed them with distilled water，and dry them for film-reading observations． The observation was about the influence of cyclophosphamide on the Mongolian gerbils' sperm． Results Compared with the negative control group，the rate of sperms abnormalities in the three CP groups showed great difference． The rate in the low-dose group and the medium-dose group was obvious different from the rate in the high-dose group( P＜0. 01 ) ． The rate in the low-dose group showed little difference from the rate in the medium-dose group( P＞0. 05) ． Morphological observation showed that:the abnormal effect of cyclophosphamide on gerbils' sperms included tail folding，tail weight / length / distortion and no hook． Conclusions Cyclophosphamide has a certain teratogenic rate on Mongolian gerbils' sperms． And it influenced their tails most． In a certain dose range，the rate of abnormalities showed little difference，while when the dose was higher than a certain level，the rate of abnormalities rose together with the dose of cyclophosphamide rose．

Abstract: Objective Methods Mensurating by Chinese Medicine Dictionary ( edition of 2005) ． Results The consequence of the 1st screening eligible shows the remarkable difference in the different seasons，different body weights，different sexes and different humidities of New Zealand Rabbits． The higher screening eligible rate of 1. 7 ～ 2. 0 kg，male New Zealand Rabbits from July to September，the suitable humidity is 61 ～ 70% ; The consequence of the 2nd screening eligible shows the remarkable difference in the different seasons，room temprature and humidity of New Zealand Rabbits． The higher screening eligible rate occurs from Jan． to Mar． ，the suitable room temperature is 22. 1 ～23. 0℃ and humidity is 61 ～ 70% ． Conclusions Under different factors，the first screening eligible rate and second screening eligible rate of New Zealand Rabbits will be influenced．

Abstract: Objective Methods cells from the bone marrow，spleen and thymus were analyzed by flow cytometry． HSCs were sorted and cultured in vitro． Results Compared with wild type mice，there are no great differences in the cell percentage and function of bone marrow，spleen and thymus in Cramp over expression mice． Conclusions Compared with the wild type mice，there was no great difference in the HSCs ` differentiation and single cell clone in Cramp over expression transgenic mice．

Abstract: Objective Methods MSCs were isolated from mice limb bone chips and in vitro culture． The proliferation and growth characteristics of MSC were observed during primary and passage cultures． Results Cells culture showed that mice bone chips derived MSCs exhibited the characteristics of mesenchymal cells． MSCs clones assay of WT mice were higher than Terc^－/－mice in primary and passage cultures ( P＜0. 01 ) ． Conclusions MSCs derived from mice bone chips can become an easy and reproducible method that does not require depletion of hematopoietic cells by sorting or immunomagnetic techniques ． MSCs of short telomere mice( Terc^－/－mice) has a more feeble feasibility in cells proliferative ability than WT mice as the seed cells for tissue engineering reconstruction．

Abstract: Objective Methods Weights data of brain，spleen，heart，lungs，liver，kidneys，adrenal glands，testis and ovaries from control group SD rats with 13 weeks，26 weeks，52 weeks，78 weeks and 104 weeks ages was obtained． These weights data and their coefficients were analyzed． Results Weights of brain，spleen，heart，lungs，liver，kidneys，adrenal glands and ovaries were increased in female rats from 13 to 104 weeks． Weights of brain，spleen，heart，lungs，liver and kidneys of male rats were higher than that of female rats，whereas adrenal weights，its organ weight / body weight ratio and organ weight / body weight ratio were significantly higher than that of males from 13 to 104 weeks． Conclusions This study firstly established different week ～ old SD rats organ weight background data and their reference range which in line with experimental animal status in our country and summarized their changes trend．

Abstract: ObjectiveM26I，and provid a basis for further study on the relationship of DJ-1 ^M26I mutation and cell proliferation and apoptosis and establish transgenic animal model． Methods Using site-directed mutagenesis kit to make the 26th amino acid mutated，constructed pcDNA3. 1 /myc-His-DJ-1 and pcDNA3. 1 /myc-His-DJ-1 ^M26I recombinant expression vectors．And then transfected them into NIH3T3 cells respectively using lipofectamine． Cells were selected with G418 at the level of 500μg /ml． Stable clones were identified on the DNA，RNA and protein levels． Using MTT assay and AnnexinV-FITC kit to detect the stable cloned cells’ viability and apoptosis level． Results NIH3T3 cells transfected with recombinant plasmid pcDNA3. 1 /myc-His-DJ-1 or pcDNA3. 1 /myc-His-DJ-1 ^M26I screened by G418． We obtained 1 and 3 positive clones of pcDNA3. 1 /myc-His-DJ-1 and pcDNA3. 1 /myc-His-DJ-1 ^M26I transfected cells by PCR detected． The Results of RT -PCR and western blot also showed the expression of DJ-1-His in pcDNA3. 1 /myc-His-DJ-1 and pcDNA3. 1 /myc-His-DJ-1 ^M26I transfected cells． MTT assay demonstrated the NIH3T3 positive cells transfected with DJ-1 ^M26I had the lower proliferation rate than normal NIH3T3 cells( p＜0. 05) ． NIH3T3 positive cells carrying the DJ-1 genes showed no significant difference compared with the normal cells NIH3T3 cells． Apoptosis test indicated that the apoptosis rate of DJ-1 ^M26I transfected cells was higher than normal NIH3T3 cells，however the apoptosis rate of the DJ-1 transfected cells was lower than normal NIH3T3 cells( p＜0. 05) ． Conclusions Recombinant expression vectors pcDNA3. 1 /myc-His-DJ-1 and pcDNA3. 1 /mycHis-DJ-1 ^M26I have constructed successfully． The NIH3T3 cells with stable expression of DJ-1 and DJ-1 ^M26I have been constructed successfully． DJ-1 ^M26I mutation can result in the apoptosis of NIH3T3 cells easily ．

Abstract: Objective Methods MSCs of Wistar rats were isolated and cultivated by bone marrow adherent culture． The cells between the third and fifth passages were prepared for transplantation． 20 male Wistar rats were divided into four groups by the number of MSCs transplanted intravenously: the 5 × 10⁵group、the 1 × 10⁶ one、the 5 × 10⁶ one and the control one ( n= 5) ． RVSPs were measured at the time of pro-transplantation，5 min，30 min and 24 h post-transplantation． The other 40 rats were randomly divided into four groups: ①the MCT /MSCs 5 × 10⁵ group; ②the MCT /MSCs 1 × 10⁶ one; ③the MCT one; ④ the control one． Rats were injected intraperitoneally with 60 mg·kg 1 MCT，except the control group with physiological saline． Phosphate buffered saline with 5 × 10⁵ MSCs and 1 × 10⁶ MSCs were transplanted into rats through external jugular vein in the first two groups respectively． At the forth week after cell administration，RVSPs were measured and lungs’tissues were stained with Hematoxylin-eosin staining，lichen dyed red staining and smooth muscle Actin immunohistochemistry． Lungs’pathological changes were observed． All data were analyzed statistically by one way analysis of variance using SPSS 11. 0． Results It was safe to transplant MSCs less than 1 × 10⁶ intravenously． At the forth week after MSCs administration，RVSP and ventricular ratio significantly decreased in the MCT /MSCs 1 × 10⁶ group compared to that in MCT group ( 47. 2 ± 10. 5 mmHg vs 35. 8 ± 8. 4 mmHg，0. 3572 ± 0. 0923 vs 0. 4454 ± 0. 0935，respectively，both P＜0. 05) ，but they didn’t differ statistically in MCT /MSCs 5 × 10⁵ group compared to in the MCT one． By pathological staining，medium thickness of the pulmonary arterioles was observed comparatively thinner in the MCT /MSCs 1 × 10⁶ group than those in the MCT group． In contrast they had no significant difference between in the MCT /MSCs 5 × 10⁵ group and in the MCT one ( P＞0. 05 ) ． Conclusions Intravenous MSCs administration can prevent lungs injury induced by monocrotaline and 1 × 10⁶ MSCs administration has better effects than 5 × 10⁵ MSCs’．

Abstract: Objective Methods Experimental mouse were exposed to sodium fluoride ( NaF) added to drinking water，The histopathological changes of testis tissue were observed by HE staining． The apoptosis were detected by TUNEL ( Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling) apoptosis Detection Kit． Results TUNEL-positive cells could be detected in fluoride-treated rat testis．The apoptotic rates of fluoride-treated testis cells were higher than that of control significantly． Conclusions There is tendency of apoptosis in testis of fluorosis in mouse． It is most similitude with change spot of pathology． There may exist apoptosis mechanism in testis lesion of fluorosis．

Abstract: Objective Methods Electrocardiogram ( ECG) and autonomic activities were recorded before，during and after ingestion 0h，2h and 4h，from conscious and unrestrained miniature swine using a non-invasive telemetry system and the autonomic nervous function was investigated by power spectral analysis of heart rate variability ( HRV) ． Results Compared with before ingestion ( BI) ，the Heart rate，autonomic activities and normalize low frequency ( LFnu) power were obviously increased，R-R interval，total power ( TP) ，very low frequency ( VLF) power and high frequency ( HF) power were significantly decreased，the LF to HF ratio was raised during ingestion ( IP) ． After food，the HR，autonomic activities and LFnu were decreased while RRI，TP，VLF and HF were increased and the LF to HF ratio was lowed，especially after ingestion for 2h and 4h． Correlation analysis showed that feeding behavior and TP，VLF，HF，LF and LF /HF were closely related，and the multivariate linear gradual regression analysis also showed that feeding behavior and the VLF，LF /HF and TP were closely related，and VLF played a major role in feeding behavior． Conclusions Feeding behavior was not only affected heart activity in miniature swine，but also could affect cardiac autonomic nervous control，which VLF played an important role in the process of feeding behavior．

Abstract: Objective Methods Twenty db / db mice aged 4 weeks were randomized to pioglitazone group ( PIO group) and db / db group，fed with 10mg / kg·d pioglitazone and placebo respectively． Ten age-matched db /misty mice were concurrently treated with placebo as non-diabetic control group ( db /m group) ． Body weight and blood glucose were measured every week． After 4 weeks' treatment，the expression of PTP1B in muscle of mice were determined with Western blotting． Results The expression of PTP1B in muscle of db / db mice significantly increased compared with db /m( P＜0. 05) ． Pioglitazone treatment significantly decreased fasting blood glucose and HOMA-IR． The expression of PTP1B in muscle of pioglitazone-treated mice obviously decreased compared with db / db group ( P＜0. 05 ) ． Conclusions Pioglitazone treatment could improve insulin resistance possibly by reducing the expression level of PTP1B in the muscle．

Abstract: Objective Methods A total of 80 Wistar male rats were randomly divided into control group and early quitting smoke groups on the smoking cessation stage 0 day，1 week，2 weeks，4 weeks，6 weeks，8 weeks，12 weeks． Levels of IL-8 protein in serum of different group rats were detected by enzyme-linked immunosorbent assay ( ELISA) ，expressions of NF-κB p65 in lung tissue were detected by S-P immunohistochemistry，and pathology score of inflammation on airway were carried out by observing HE stained lung tissue sections under light microscope． Results Early quitting smoke group could be seen Cilia sticking together，lodging，epithelial cell vacuoles deformation，necrosis，hyperplasia，inflammatory cell infiltration． The levels of IL-8 protein in serum，expression of NF-κB and pathological score in lung tissue of smoking cessation groups on any smoking cessation stage was significantly higher than non-smoking control group，statistically significant ( P＜0. 05) ． In early quitting smoke groups，the serum level of IL-8，expression of NF-κB and airway inflammation pathological score of groups was the highest after 8 weeks of smoking cessation，and then the serum level of IL-8 tended to decrease and the lung inflammation alleviated on the smoking cessation stage 12 weeks． Conclusions Although on the early stage of smoking cessation，inflammatory response can be seen a slight increase，but with the cessation prolonged inflammatory response can still be seensome relief． Therefore，it is necessary to persist on quitting smoking as soon as possible

Abstract: Objective Methods Mouse MIP-1α and B7-1 genes were synthesizeed and amplification by PCR．Target genes were directly connected with Lentivirus vector，the production of which were transformed into Bacterium coli DH5α cells，and the positive colones were identified by PCR and direct sequencing analysis． Then the plasmids of MIP-1α and B7-1 genes infected 293T cells，respectively，green fluorescence protein ( GFP ) in 293T cells was observed by fluorescence microscope; Western Blot was used to test protein expression of MIP-1α and B7-1 genes and Real-time PCR was used to detect the titer of lentivirus． Results Lentivirus-mediated mouse MIP-1α and B7-1 gene vectors were successfully constructed and the titer of which was 2. 00E + 8 TU /ml tested by real-time PCR． Conclusions Lentivirusmediated mouse MIP-1α and B7-1 gene vectors were successfully constructed and lay a foundation for gene therapy with lymphoma in the future．

Abstract: Objective Methods 22 male Kun Ming mice were divided into the normal control ( NC) group and CIH-CR group． The mice of CIH-CR group suffered 8 hours intermittent hypoxia everyday for 4 weeks． The concentration of O₂ and CO₂ in the cabin was detected by oxygen analyzer and Carbon dioxide detector，meanwhile，the blood oxygen saturation were detected by blood oximeter． At the end of the experiment，weight of right and left ventricles was measured． The general pathological changes of myocardium，pulmonary，kidney and brain were observed by HE staining． Result The concentration of O₂ and CO₂ in the cabin and the SO₂ in the empennage of mouse accord with the cyclical change process． Compared with the NCgroup，the right ventricular hypertrophy were increased in the CIH-CR groups( P＜0. 01 ) ． The tissue of myocardium，pulmonary，kidney and brain were showen obvious hypoxic change in CIH-CR group． Conclusions A CIH-CR animal model were be established successfully．

Abstract: Objective Methods Rats of model groups were given silaenafil 10mg / kg by gavage． Rats of control groups were given normal saline only． All rats were gavaged once a day for two weeks． Rats were observed for 2 hours after each drug administration for penile erection，genital grooming and possible copulation mounting． The number of responding rats was recorded along with the number of sexual activity episodes ( penile erection，genital grooming or copulation mounting) ． Penile erection index ( PEI) was calculated for each group by multiplying the percentage of active rats ( responding rats) by the total number of activity episodes． By using the modified assessment method，male rats were observed by camera． Meamwhile，we also stimulated the male rats by female rats at different periods． Results Among female rat-stimulated groups，the model groups compared with the control groups，PEI was significantly increased ( P＜0. 05) ． Among the model groups，the camera observed group compared with the human observed group， PEI was significantly increased ( P＜0. 05 ) ． Compared with non-female rat-stimulated group，PEI was significantly increased in female rat-stimulated group( P＜0. 05) ． The 2 h PEI showns no significant differences between sustained induction group and intermittent group． But，during the period of the second hour，PEI was significantly increased in intermittent group ( P＜0. 05 ) ． Conclusions After optimized，the assessment methods can evaluated the strengthening yang efficacy of candidate drugs more effective．

Abstract:As a tool of genetic engineering，DNA transposons have been widely used for transgenesis and insertional mutagenesis in various organisms． Till now，transposons active in mammalian include: 1) hAT-like Tol2; 2) ，two Tc1-like transposons，Sleeping Beauty ( SB) and Frog Prince ( FP) ; 3) PiggyBac family． Among those transposons， the moth-derived transposon piggybac appears to be the most promising genetic tool due to its higher efficient transposition and higher cargo capacity． Therefore，it can be used for transgenesis，discovery of cancer gene and tumor suppressor gene，and gene therapy． The traceless excision transposition property of the PB transposon can be used to generate transgene-free iPS cells while maintaining an unaltered genome． In this review we will discuss the application and the future of PB transposon in mammalian．

Abstract:Eye irritation test is a major field for cosmetics safety assessment． Many different in vitro methods were designed to replace traditional animal testing，these methods are in the process of development，validation and acception with different stages． It is no single in vitro methods can completely replace the rabbit eye test，but with reasonable evaluation the scientific principle，relevance and standardization of alternative methods，which will contribute to select and guide the application of alternative methods including mechanism study，classification and labeling，quality control and product development for cosmetics ．

Abstract:The characteristic of autoimmune diseases is the loss of B cell tolerance，BAFF plays an important role in B cell proliferation and survival by binding to its receptors． BAFF-transgenic mice is likely to develop autoimmune diseases． So BAFF antagonism or inhibition may be a target for the treatment of autoimmunity． In this paper，we summarize progress in understanding the role of BAFF in autoimmune diseases．