Abstract:Objective To investigate the regulation role of gastric cancer related miR-148a on gastrin receptor CCKBR expression, and find the correct binding sites of miR-148a in CCKBR 3'UTR. Methods The potential binding sites of miR-148a in the CCKBR 3'UTR were predicted with the bioinformatic tools; The miR-148a expressing plasmid was constructed by PCR, and miR-148a expression was verified by Northern Blot; The luciferase report plasmids containing the wild type and mutated binding sites of CCKBR 3'UTR were constructed, and were used to study the regulation mechanism and identify the binding sites of miR-148a by luciferase activity analysis; The regulation effect of miR-148a on CCKBR protein expression was checked by Western Blot. Results Three potential binding sites of miR-148a in the CCKBR 3'UTR were found; The miR-148a expressing plasmid was constructed successfully, and highly expressed miR-148a after transfected to gastric cancer cells; The inhibitory effect of miR-148a on CCKBR protein expression was checked by Western Blot. Over-expression of miR-148a inhibited CCKBR expression by directly binding to the binding site in CCKBR 3'UTR 423bp. Conclusion CCKBR is a target of miR-148a, and its expression is inhibited by the binding of miR-148a on its 3'UTR, indicating that miR-148a may participates in the progression of gastric cancer by regulating CCKBR expression.

Abstract:Objective To investigate the effects of different doses Alpha-Linolenic acid (ALA) on the expressions of fatty acid synthesis-related genes in HepG2 cells. Methods HepG2 cells were divided into two groups, normal control group (NC) and high fat group (HF) in which cells were firstly cultured for 36h in the medium contained 0.5mmol/L stearic acid. Real-time quantitative PCR was taken to detect mRNA expression of genes SREBP1C, FAS and ACC which are related to fatty synthesis. While there are significant differences in fatty synthesis, 10%, 20%, 50%, 70% and 100% ALA took the place of stearic acid, 36h later, real-time quantitative PCR and Western blotting were taken to detect mRNA and protein expression of genes related to fatty synthesis. Results SREBP1C mRNA expression of ALA substitution groups were significantly lower than the high-fat group (P< 0.001).The FAS of 0.5 mmol/L ALA group and 0.35 mmol/L ALA group were significantly lower than the high-fat group (P < 0.001). ACC genes mRNA level was not significantly different from high-fat group. SREBP1C and FAS protein expression were significantly lower than the high-fat group, but ACC showed no significant difference. Conclusions Saturated fatty acids promote hepatocyte fatty acid synthesis, ALA abates fatty acid synthesis by inhibiting FAS and SREBP1C gene expression.

Abstract:Objective To investigate the effects of morroniside on the expression of Wnt7a and APC in rat cortex of focal cerebral ischemia/reperfusion. Methods After middle cerebral artery occlusion (MCAO) model was performed SD rats were divided into groups of sham, model, and morroniside (30 mg/kg, 90 mg/kg,270 mg/kg). The expression of Wnt7a and APC were evaluated by Western blot. Results Compared with the sham group, the expression of Wnt7a in rats of MCAO model was increased, and the expression of APC was decreased. Compared with model group, morroniside at the concentrations of 30 mg/kg, 90 mg/kg, 270 mg/kg decreased APC expression respectively, while 270 mg/kg morroniside-treatment obviously increased the expression of Wnt7a. Conclusion Morroniside could increase the expression of Wnt7a, and inhibit the expression of APC after ischemic injury, therefore activating the Wnt signaling pathway.

Abstract:Objective To study the effect of Xi Yanping injection on electrocardiogram in Beagle dogs. Methods 24 Beagle dogs were divided into negative control group (0 mg/kg), low-dose group (15 mg/kg), middle-dose group (75 mg/kg), high-dose group (250 mg/kg).Each group consist of 3 animals/sex/group, all animals were administrated by intravenous drip at 20 mL/kg once a day. Administration period was 28-days with 14-days recovery phase. Animals LeadⅡ electrocardiogram (ECG) was measured during quanrantine, administration and recovery phase. The following parameters included heart rate, P -R interval,Q - T interval, P, Q, R S wave amplitude and ST and T wave amplitude were determined, serum biochemical parameters were measured to evaluate the effect on main organs. Results All parameters of quarantine period were fell in the normal limit. After the first administration, heart rate decrease was found in low dose group and P-R interval prolongation was found in high dose group. No abnormal was found in dose metaphase,end of dose stage and recovery stage. Conclusion Xi Yanping injection had no obvious influence on ECG of Beagle dog.

Abstract:Objective To study the expression of divalent metal transporter 1(DMT1)and ferroportin 1(FPN1)in obese mice's duodenal epithelium and investigate the mechanism of the effect of obesity on iron absorption in mice. Methods C57BL/6J mice were randomly divided into control group and obesity model group, each group of 6, To establish obese mice model by having a high-fat diet and the control group were fed with a normal diet for 12 weeks. After completion of modeling, The level of DMT1 and FPN1 mRNA expression in the duodenum were measured by real-time fluorescent quantitative PCR(Real-time PCR) method, the protein expression of FPN1 was measured by Western-Blot. Results Compared with the control group, the level of DMT1、FPN1 mRNA and FPN1 protein expression in the duodenum were decreased significantly in obese mice(P < 0.05). Conclusion Obesity can decrease the expression levels of DMT1、FPN1 mRNA and FPN1 protein and induce iron deficiency,in order to provide experimental and theoretical basis for studying the mechanism of iron deficiency caused by obesity further.

Abstract:Objective To establish a rapid modified Local lymph node assay (LLNA) for the sensitization and irritancy potential of chemicals, and evaluate cosmetics products. Methods 3 kinds of chemicals (2, 4-Dinitrochlorobenzene (DNCB), Eugenol and Hexyl cinnamic aldehyde) and 3 kinds of cosmetics were selected. The dorsal sides of both ears of female BALB/c mice were treated with 25μL/ear of the test solutions on 3 consecutive days, and the thickness of each ear was measured. BrdU solution was injected inter-peritoneally on day 5. On day 6, each auricle was weighed, the bilateral draining auricular lymph nodes were excised and weighed, then made into single cell suspension. BrdU was measured by ELISA using a commercial kit. Results Ear swelling and auricle weight of DNCB (1.0%), Eugenol (25% and 50%) and number 3 cream foundation increased significantly (P < 0.05 or P < 0.01), which may be considered as irritants. The others were negative. For LLNA, three chemicals and number 3 cream foundation pretended positive. Conclusion The combined LLNA/ear swelling/ear weight assay appears to be a reliable predictor of sensitization and irritancy potential, and is expected to play an important role in safety evaluation of cosmetics.

Abstract:Objective To study the main pharmacodynamics effect of compound lavender ointment on the scald,anti-inflammation and analgesia. Methods Sixty guinea pigs were divided into normal groups,model groups(superficial Ⅱ°scalding models),compound lavender ointment high,median,low dose groups,and jing wan hong ointment groups,then observation of the wounds and healing was made and recorded. Histopathological examination was made to examine the wounds,and ELISA to analyze serum TNF-α、IL-6、IL-8. Xylene ear swelling model and writhing test were used to observe the analgesic and anti-inflammatory effects of the compound lavender ointment. Results Compound lavender ointment can relieve the wounds swelling and exudation,which may be related to the decrease of proinflammatory cytokines(P< 0.01),and promote the wound healing significantly(P< 0.01),that the side-effects were not observed. High dose of compound lavender ointment can inhibit ear swelling and have certain analgesia with the writhing tests. Conclusion Compound lavender ointment have the promoting wound healing,anti-infectious and antalgic effects on the scald.

Abstract:Objective To provide an experimental reference for clinic application of Jiazhu vascular ligating clips on ligation effects and the impact on the body in rabbits after laparoscopoic cholecystectomy. Methods 36 rabbits were randomly divided into 3 groups, normal control group, Jiazhu group and similar product control (Hem-o-lok) group, 12 rabbits in each group. Except for normal control group, the rabbits were used gallbladder duct and gallbladder artery closed by vascular ligating clips and Hem-o-lok clips in Jiazhu group and similar product control group, respectively. The changes of blood biochemical, electrolytes, hematological parameters, coagulation, ligation effects and organ coefficients were observed at post-operation during 12 months. Results Compared with normal control group, the rabbit weight was reduced in the Jiazhu group at 3 days postoperatively (P< 0.01), ALT level was increased at 1week postoperatively (P< 0.05), CREA and ALB levels were all significant decreased at 1～2 weeks postoperatively (P< 0.05, P< 0.01), NEUT numbers and TG content were significant increased at 2 weeks postoperatively (P< 0.05), GLU content was significant decreased at 2 weeks and 1 month postoperatively (P< 0.01); LYM and RBC numbers were significant decreased at 1 month and 1 week postoperatively (P< 0.05), respectively. PLT numbers were significant increased at 1～2 weeks postoperatively (P<0 .01). While the rabbit weight was reduced in the Hem-o-lok group between 3 days and 1month postoperatively (P< 0.05, P< 0.01), and ALT level, NEUT and PLT numbers were increased at 1 week postoperatively (P< 0.05, P< 0.01), ALB and GLU contents were all significant decreased at 2 weeks postoperatively (P< 0.01), TC content was significant increased at 1～2 weeks postoperatively (P< 0.05). The rabbit weight, blood biochemical and hematological parameters in the Jiazhu group and Hem-o-lok group were all no significant differences with normal control group in the remaining time (P> 0.05), and there were all no significant differences with electrolyte, coagulation and organ coefficients between each group (P> 0.05). Conclusion Jiazhu vascular ligating clips are safe and reliable to close gallbladder duct and blood vessels, It shows that there are no differences in the validity and security between Jiazhu vascular ligating clips and Hem-o-lok clips.

Abstract:Objective To select a best mouse strain for establishing an animal model of orthotopic transplanted liver cancer through comparison analysis.Methods C57,C3H and BALB/c mice(each 10)were selected as model groupⅠ,Ⅱ and Ⅲ.H22 cells were inoculated into the liver parenchyma of mice in model group along the ventral midline laparotomy.Fifteenth days after injection of laparotomy,the rate of tumor was observed,the volume of ascites and gross tumor volume were measured,and tumor pathology was analyzed.Results The survival rate of mice in three groups was 100%.After 15 days,ascites in three groups were produced,but with no significant difference among the three groups of the volume of ascites.Tumor transplantation rate in model groupⅠ was 100%,which was higher than 60% in model group Ⅱ and 30% in model group Ⅲ.Liver tumors in model group Ⅰ were all large compact grayish white lesions,and the average tumor volume was significantly greater than that of model group Ⅱ and Ⅲ(P< 0.05). Pathology results confirmed that white lesions in liver of three groups were hepatocelluar carcinoma in situ.Conclusion C57 is the better ideal experimental animal for establishing orthotopic liver caner model,and these results will provide a good experimental platform for research in pathogenesis of hepatocellular carcinoma.

Abstract:Objective To explore the basic ingredients of the tree shrew's(Tupaia belangeri)milk and compare with the dairy ingredients of other milks. Methods We select ten seed tree shrews after delivery (1 ～ 21) d with lactation mother tree shrews, and use artificial passive breastfeeding method let the young tree shrews suck breast milk,we took the milk from the young tree shrews in the stomach, directly using aseptic operation with a syringe immediately, once every two days, for consecutive three to five times, and a total of 18 mL milk was taken from each seed tree shrew. Then the milk was detected according to the national standard method for component testing. Results The total solid content of the tree shrew's milk was 43.63%, including 26.01% of fat, 10.41% of protein, 0.45% of lactose and 0.99% of ash content. Compared with cow's milk, the tree shrew's milk contained 3.36 times of total solid contents, 1.24 times of ash, 2.74 times of protein, 6.67 times of fat, and 0.09 times of lactose.Compare with baby formula milk, the tree shrew's milk contained 1.44 times of total solid contents, 0.20 times of ash, 0.58 times of protein, 1.53 times of fat, and 0.06 times of lactose. The trace mineral composition of the tree shrew's milk showed that the calcium, phosphorus, potassium, sodium, magnesium, and iron contents were 1.83 times, 2.73 times, 1.25 times, 1.93 times, 1.28 times, and 1.48 times higher than those in the cow's milk, and were 0.66 times, 0.85 times, 0.34 times, 0.26 times, 0.85 times, 0.24 times lower than those in baby formula milk. Conclusions The main nutrients of tree shrew's milk is of high fat, high protein and low sugar, and it can provide a basis for tree shrews artificial brood and breeding work.

Abstract:Objective To study the effects of toys for laboratory animals on reproductive performance and growing development of experimental mice. Methods ICR mice were fed with toys, the informations of reproductive performance and growing development were recorded. Results All the data of reproductive performance of the test group were higher than the control group except the number of newborn mice, and showed significant difference (P< 0.05). The data of growing development of the test group were higher than the control group too, but showed no significant difference (P> 0.05). Conclusion Toys for laboratory animals have good effects on reproductive performance and growing development of mice, and suggested to be used into the process of experimental mice raising.

Abstract:Objective The purpose of this study is to select suitable ages of rats for the CPR (cardiopulmonary resuscitation) animal model. The neurological function score and subgroups analysis are evaluated in 2 month old and 4 month old animal groups. Methods Based on the evaluation of physiological indexes including ECG, blood pressure and neurological function defect score (NDS) and subgroup analysis, the stability of CPR rats model was compared between 2 month old and 4 month old animal groups. Results The results showed that, the model rate of the ventricular fibrillation was induced by electrical stimulation, the 4 month old group was 87.5%, significantly higher than the 2 month old group, however, there was no significant difference between the two groups in the mortality rate; For the changes of blood pressure during the process of CA(cardiac arrest) induced by electrical stimulation, the 4 month old group was significantly lower than the 2 month old group (P< 0.01); for the NDS at each time point after CPR, there was no significant difference between the two groups; however, the NDS subgroup analysis at different time points showed that there were different degrees of differences between the two age groups (P< 0.05). Comparing with the 2 month old group, the 4 month old group had a stable process during the animal model preparation, had an obvious cerebral blood hypoperfusion phenomenon and aggravation of brain injury after CPR. Conclusion The 4 month old rats are more suitable for preparation of CPR animal mode, the model rate is high, the brain injury aggravate. It is more suitable evaluation for basic research and treatment of CPR.

Abstract:Objective To identify and compare melanin distribution in iris and skin tissues of rabbits from four breeds including WHBE rabbit. Methods Ferrous sulphate staining was performed for iris and skin tissue of rabbits from different breeds. Results It showed dark green staining in both iris stroma and iris pigment epithelium of WHBE rabbit, while light green staining was observed in its follicles. Conclusion The results of histologic evaluations provide some clues to the future research on mechanism of WHBE rabbit phenotype mutation.

Abstract:Objective To open pterygopalatine artery using autologous blood clot clogged the middle cerebral artery, resulting in cerebral infarction model of middle cerebral artery in the distal blood supply area. Methods Open PPA, ligation of ECA, embolus through CCA blood flow, rushed into the MCA distal embolus, cause infarction model. According to Bederson rating score the neural function. TTC staining to determine the infarction volume. Results Comparison of two operation models, scores were increased, and white infarcted area appears. There is no significant differences detected by statistical analysis. Conclusion Open PPA manufacturing cerebral infarction model rats of autologous blood clots could shorten the operation time, reduce the mortality rate.

Abstract:Vascular dementia is one of the common causes of cognitive impairment. Occlusion of bilateral common carotid arteries, middle cerebral artery occlusion, as well as induced hypertension are major methods to make rat model for vascular dementia. The present review will summarize research progress in medical and non-medical treatment of vascular dementia rat models in order to provide theoretical direction for clinical practice.

Abstract:Metagenomics (metagenome) obtained all the microorganisms genome in specific biological environment from DNA samples directly from soil,water and skin,body cavity of human and animals. The metagenomic library was constructed by cloning them into alternative host cells using the vector,which is a discipline of screening new active substances and new genes by high-throughput sequencing. Therefore,using metagenomics not only can effectively detect the composition of microbial community of specific environment,but also greatly expanded the utilize the space of the microbial resources,developed of the next sequencing, enriched the content of bioinformatics. Based on the advantages of metagenomics,we summarized the major advances of metagenomics in human and animal pathogenic microorganisms in recent years,which might be used for the investigation and analysis of microbial pathogens of laboratory animal, animal epidemic monitoring and diagnosis of biological safety.

Abstract:Laboratory animal science teaching has become an indispensable part of medical education. This paper summarized teaching characteristics of laboratory animal science at different levels, including the curriculum setting background, curriculum arrangement as well as assessment methods, in order to provide some references for the teaching staff of laboratory animal science.