Abstract:Objective To investigate the effect of gut bacteria under chronic colitis on the progression of hepatoma in mice. Methods 22 hepatitis B virus (HBV)-transgenic mice (male, 8 weeks) were randomly divided into two groups, one group (n=10) was fed the drinking water containing 2% dextran sodium sulphate(DSS)to induce chronic colitis and the control group(n=12)was fed with normal drinking water. In order to investigate the effect of gut microbes, 7 male HBV-transgenic mice(8 weeks, with no detectable hepatoma under microscopy) were cohoused with 4 mice with hepatoma for 16 weeks. Results No significant liver cell damage was observed in the group of the mice fed with 2% DSS-containing drinking water. By the 22-week old, 9 of the 10 mice(90.0%) fed with 2% DSS-containing drinking water, 2 of the 12 mice(16.7%) fed with normal drinking had hepatoma. Both the hepatoma incidence and the tumor numbers in the group of mice fed with DSS-containing water were significantly higher than that in the controls (P=0.002 and P=0.028respetively). Compared to controls, the bacteria family Prevotella (P=0.022) and Anaeroplasma (P=0.014) reduced significantly in the mice with induced chronic colitis. All the mice (n=7) cohoused with the mice with hepatoma had the liver tumor developed at 24-week-old. Conclusion Alterations of gut bacteria under chronic colitis may promote the development of liver cancer.

Abstract:Objective To detect the expression change of ETFβ in diabetic nephropathy rats and study the role of ETFβ in fatty acid-induced apoptosis in renal tubules. Methods Diabetic nephropathy model was established by intraperitoneal injection of streptozotocin and unilateral nephrectomy. In vivo ETFβ expression was detected in renal cortex, as well as tubular injury evaluated. In vitro fatty acid-induced apoptosis in renal tubular cells NRK 52E model was established and ETFβ recombinant plasmid was constructed to be transfected into NRK 52E cells and furtherly to observe the effect of ETFβ over-expression on the fatty acid-induced apoptosis. Results In the rats model of diabetic nephropathy induced by streptozotocin injection and unilateral nephrectomy, ETFβ mRNA and protein expression were decreased as obvious tubular damage occurred. Fatty acids could induce apoptosis in NRK 52E, and ETFβ over-expression reduced the apoptosis. Conclusion The expression of ETFβ is decreased in diabetic nephropathy model, and ETFβ over-expression can reduce apoptosis induced by fatty acid in renal tubular cells.

Abstract:Objective To assess the protective of Curcumin and Tea Polyphenols against different time of UVB-induced acute photo damage in hairless mice skin. Methods Thirty six BALB/C hair1ess mice were randomly divided into control groups and treatment groups(all of them were divided into 0 s, 30 s, 60 s, 120 s, 240 s groups with two hairless mice).Curcumin and Tea Polyphenols were applied to the mice’s skin before 30min irradiation. The irradiation distance was 15cm. The mice in each group were given three times of UVB MED about 200～540 mJ/cm² and the irradiation time were 30 s、60 s、120 s、240 s. After irradiation, the skin on the back of the radiation center were taken into paraffin section for light microscopy observation. Results Different amount of squamae appeared in the control group of hairless rat, and lymphocytes scattered in collagen tissues. In the 120 s and 240 s UVB irradiated group, the collagen was red stained, disorderly arranged, even homogenized. In the treatment group, inflammatory cell infiltration was lessened, and collagen damage weakened. Conclusion By weakening inflammatory cell infiltration and collagen damage, topical application of Curcumin and Tea Polyphenols can protect hairless mice from acute photodamage caused by UVB irradiation.

Abstract:Objective To establish a rat model at the same time in accordance with the "hot sheng syndrome" of traditional Chinese medicine and primary immune thrombocytopenic purpura of peripheral blood platelet reduction. Methods Using back multi-point injection of 20% dry yeast suspension on SD rats and 1:4 dilution of rabbit anti SD rats platelet serum (APS) by intraperitoneal injection to establish a primary immune thrombocytopenic purpura "heat sheng" rat model. And observing rats of TCM syndrome characteristics, hemogram, myelogram and serotonin (5-HT) level of the temperature regulating center in thalamus. Results After injection of 2 h ～ 6 h temperature and daily water of the model group rats increased significantly, toe purper showed in fourth day of modeling and intestinal mucosal bleeding in thirty day of modeling(P < 0.05);Platelet count in peripheral blood decreased significantly, bone marrow megakaryocyte number reduced significantly((P < 0.05);5-HT level of the temperature regulating center of brain increased significantly((P < 0.05). Conclusions The study of the primary immune thrombocytopenic purpura heat sheng rat model of combination of disease and syndrome reflected basically the pathological characteristics of purpura caused by "heat sheng" in primary immune thrombocytopenic purpura rat mode.

Abstract:Objective to provide the evidence for inducing the SAP model in rats with proper concentration of sodium taurocholate. Methods 60 SD rats were divided into sham operated group, group of 1.5% in concentration, group of 3.5% in concentration and group of 5% in concentration randomly, while the SAP model was induced by the sodium taurocholate concentration of 1.5%, 3.5% and 5% with the method of retrograde injection into the biliopancreatic duct.To calculate the mortality of different groups, measure the serum amylase, tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6), and to observe the pancreatic pathological scores of HE staining in rats. Results The mortality in group of 5% in concentration has a significant ascending compared with group of 1.5% in concentration, while the serum amylase, tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), pathological score of hemorrhage and acinar necrosis in group of 5% in concentration have a significant ascending compared with group of 1.5% in concentration and group of 3.5% in concentration. Conclusions A better SAP model may be induced by sodium taurocholate with the concentration of 5% by the method of retrograde injection into the biliopancreatic duct, which may accord with the physiological and pathological manifestation of SAP.

Abstract:Objective To determine whether dopamine D5 receptor (D5R) regulates the development of dilated cardiomyopathy (DCM) by inhibiting oxidative stress. Methods We developed heart-specific hD5 mutant (α-MHC-hD5F173L) and wild type (α-MHC-hD5WT) transgenic mice. The NOX2 expression and ROS production were tested in the transgenic mice at three month of age. The α-MHC-hD5F173L mice were treated with either NADPH oxidase inhibitor Apocynin (1mmol/kg/day) or phosphate-buffered saline (PBS) as control by intraperitoneal injection for 4 weeks. After then, the indexes of heart function were measured. The hD5WT and hD5F173L were transfected respectively in rat H9C2 cells, in which ROS production and NOX2 expression were detected at basal level. Results The ROS production and NOX2 expression were higher in the heart of α-MHC-hD5F173L than α-MHC-hD5WT mice. Apocynin treatment improved the heart function of α-MHC-hD5F173L mice. NOX2 expression and ROS production were higher in hD5F173L than hD5WT transfected H9C2 cells. Conclusions Dopomine D5 receptor may prevent DCM development by inhibiting oxidative stress.

Abstract:Objective The aim of this study is production of organ specific animal model for studying reproductive toxicity in mice. Methods F1 generation was gotten by mating the Ddx4-cre transgenic male mice with the Rosa26mT/mG transgenic female mice. F1 offspring and its parents phenotype was screened by molecular biological, histopathological and in vivo imaging technology. Results At molecular level, specific DNA fragment was only found in testis of F1 offspring; At the organ level, the expression of green fluorescent protein could only be observed in testis of F1 offspring; Testicular frozen sections and sperm fluorescence observation showed that green fluorescent protein were mainly expressed in the germ cell lineage such as secondary spermatocyte and spermatocyte and spermatozoon. Conclusions The production of the mice with specific germ cell expressed green fluorescent protein by Cre/loxP recombination system were built successfully.

Abstract:Objective To analyse and monitor the genetic quality of closed colony NIH mice in Beijing district for the last 3 years. Methods We use biochemical genetic markers(including alkaline phosphatase-1 and the like 14 biochemical markers), selecting A and B colonies from different facilities for genetic monitoring in 2011 to study the polymorphism. And in 2014, 30 NIH mice just from B colony were monitored using the same testing and sampling methods. Results In 2011, NIH mice form both A and B facilities existed 6 polymorphic biochemical markers(Ce2, Car2, Gpi1, Es10, Gpd1, Pgm1); and NIH mice of B company also existed polymorphism in Es3 loucs. Between the 2 NIH mice colonies, there were significant difference in Es3、Gpd1、Pgm1 loci (P < 0.01), and difference in Car2 locus(P < 0.05). FST of the 2 colonies was 0.0406, the genetic identity was 0.9619, and the genetic distance was 0.0388. In B company, NIH mice of 2014 appeared 2 homozygous loci(Ce2 and Gpd1) when compared with NIH mice of 2011. Between the 2 NIH mice colonies, there were significant difference in Es10 and Gpd1 loci (P < 0.01), and difference in Pgm1 locus(P < 0.05). Fst of the 2 colonies was 0.1103, the genetic identity was 0.8847, and the Genetic distance was 0.1266. Conclusions Population isolation, breeding and selection, populationpopulation quantityquantity and generation significantly affected the genetic architecture of NIH mice. So when breeding and reserving seeds, we should strengthen the genetic monitoring of outbred NIH mice, in order to offer reliable genetic quality protection.

Abstract:Objective To investigate the synergism and attenuation effects of Scapharca Subcrenata Extraction on NP chemotherapy in model mice. Methods The models of nude mice were induced with A549 cell line xenograft. The tumor inhibiting rates, body weight, food intake, hematology and blood biochemistry index were determined to evaluate the synergism and attenuation effects of Scapharca Subcrenata Extraction (125、250、500 mg/kg, i.g) on NP chemotherapy. Results Compared with NP chemotherapy group, the tumor inhibiting rates, body weight, food intake, white blood cell number were increased and glutamate pyruvate transaminase, glutamic oxalacetic transaminase and urea nitrogen were decreased markedly in NP chemotherapy plus Scapharca Subcrenata Extraction groups. Conclusion Scapharca Subcrenata Extraction has a remarkable synergism and attenuation effects on NP chemotherapy.

Abstract:Objective To explore effects of parents exposure to TBT on blood routine of F1 generation mice. Methods 80 mice including 40 males and 40 females, were randomly divided into control groups (CK), low dose groups (LTBT), middle dose groups (MTBT) and high dose groups (HTBT). They were given dose of TBT (0, 0.2, 2, 20μg/kg) every day. The experiment lasted 45 days. At 60 days, one female and one male of the same concentration were bred in the same cage according to 1:1. At postnatal day 60, blood was collected for the determination of blood routine. Results Compared with control group, the number of red blood cells and hemoglobin of F1 generation male mice in LTBT and HTBT groups were significantly increased (P < 0.01); Red blood cell volume, mean corpuscular hemoglobin (P < 0.01), and the lymphocyte absolute value in F1 generation male LTBT were significantly reduced (P < 0.05); HTBT of female mice were significantly increased about the number of red blood cells (P < 0.01). A dose-dependent increase of the hemoglobin, red blood cells, and platelet count of F1 generation female experimental groups was observed. Conclusion Parental TBT exposure affects the F1 mice blood routine. There is the greatest influence on LTBT in F1 generation male mice and on HTBT in F1 generation female mice.

Abstract:Objective To investigate endoscropic assist in rat orotracheal intubation. Methods 40 rats were randomly divided into two groups. The procedure time, total time, intubation frequency, the success rate of the first intubation and the survival rate in 24h were compared between the conventional group and the endoscropic group. Results The total time in two groups were nearly the same. But the procedure time and frequency of intubation were significantly less in the endoscropic group than those in the conventional group. The success rate of the first intubation in the endoscropic group was obviously higher than that of the conventional group. There was no obviously difference between the survival rate after extubation in 24h between groups. Conclusion Endoscropic assist is superior to the conventional method and worth promoting.

Abstract:Objective To improve the method of retinal vascular preparation and application in experimental animals of mice, rats, dogs and monkeys. Methods The retinal vessels were isolated with proteinase K-trypsin digestion technique. The samples were stained by hematoxylin-eosin(HE) and Periodic-Acid Schiff(PAS). The expression of CD31 and VEGF were detected by immunohistochemistry(IHC). Results The spread sheets of vascular network are complete, and can clearly show the blood vessels. No nerve tissue residue is left. Different levels of vascular morphology are clear. The vascular branches are clear and complete with no fracture. The cell morphology is intact, and the cell nucleus is clearly dispayed. The entire process can be completed in a short time, and has a high success rate. The vascular antigen are successfully retained: Expression of CD31 can be seen in both pericytes and endothelial cells; VEGF is mainly expressed in endothelial cells. Conclusions The improved method is successfully applied in different experimental animals. This approach will provide an important contribution to the retinal vascular disease research.

Abstract:Objective To analyze and compare results of local lymph node assays (LLNA) between BALB/c and CBA mice. Methods 4 chemicals (2, 4-dinitrochlorobenzene, eugenol, hexyl cinnamic aldehyde and isopropanol) and 3 cosmetics were applied to the dorsum of both ears of Balb/c and CBA mice for three consecutive days. BrdU solution was injected inter-peritoneally on day 5. On day 6, the bilateral draining auricular lymph nodes were excised and made into single cell suspension, the lymph cell proliferation was measured by BrdU ELISA kit. Results 2, 4-dinitrochlorobenzene, eugenol, hexyl cinnamic aldehyde and NO.3 perm agent pretended positive for both strains, EC1.6 values of three chemicals were found to be 0.08% (very strong), 4.02% (moderate), 6.68% (moderate) and 0.07% (very strong), 6.08% (moderate), 8.89% (moderate) for BALB/c and CBA mice respectively. Isopropanol, NO.1 and NO.2 cosmetics pretended to be non-sensitizers with SI<1.6 for both strains. Conclusion This study showed that BALB/c mouse was essentially equal to CBA for LLNA: BrdU-ELISA, which suggested that BALB/c mouse was a good alternative for CBA used in chemicals and cosmetics allergenic evaluation.

Abstract:Objective To establish the amplified luminescent proximity homogeneous assay(AlphaLISA) for the detection of Sendai virus. Methods The antigen concentration, serum concentration and the donor beads/acceptor beads ratio used in the AlphaLISA method were optimumized by the phalanx experiments, then the antibodies of Sendai Virus in 40 rat sera were detected by the established AlphaLISA method and ELISA detection method. The results were compared and the differentia between the two methods was confirmed by IFA. Results The optimum concentration of SV bio-peptide in AlphaLISA assay was 250 nmol/L, the best proportion of donor beads/acceptor beads ratio was 1:1, using the concentration of 20 μg/mL and the serum dilution was 1:10000.7 of the 40 rat sera were detected SV positive by ELISA, the positive rate was 17.5%, 8 of the 40 rat sera were determined SV positive by AlphaLISA, and the positive rate was 20.0%, the AlphaLISA positive serum was confirmed by IFA. Conclusions We preliminary established the Amplified Luminescent Proximity Homogeneous Assay(AlphaLISA) for the detection of Sendai virus. The sensitivity of the method is comparable to classical ELISA method, but this method use less serum samples and without washing steps. The method has some advantages in degeneracy and accuracy.

Abstract:Diabetic neuropathic pain(DNP)is one of the most common complications in clinical, which influenced patients’ daily functions greatly, without clear mechanisms and effective methods. P2X3 receptors play a pivotal role in the formation, transmission and conduction of pain under neuropathic pain models, associated with peripheral sensory nerve excitability enhancement. This paper focuses on the establishment of DNP models, and the effects of P2X3 receptors in diabetes mellitus.

Abstract:Objective To assess the health status of rodent laboratory animals by pathological diagnosis, our lab has being take apart in investigating the quality of laboratory animals in Beijing area for years and offer some advices for standardized breeding to ensure accurate results of scientific research. This paper focuses on the analysis of laboratory rodent samples that collected in October 2014. Methods We collected the heart, liver, spleen, lung, kidney, large intestine and small intestine, and put these organs into 10%Calcium formaldehyde solution for fixation, and then prepared into two different sections for optical microscopy observation including all paraffin specimens stained with H&E and the frozen sections stained with Oil Red-O and PAS. Results The vast majority of laboratory rodents were up to standard, but there still a problem in individual units. The main problem is liver and lung disease. The rate of Hepatocyte swellingis 6% (mouse), 2.5% (rat), 8.2% (guinea pig), moreover part of them were lipidosis, according to Oil Red-O stain. the mainly problem of lung is congestion, edema and Interstitial pneumonia, the detectable rate of pulmonarydiseases is 15.5%(guinea pig). Conclusions The vast majority of laboratory rodents were pathologically diagnosed as healthy animals.The liver disease may be caused by improper feeding.And disease of lung may led by haze, unqualified bedding and low temperature.

Abstract:Objective To provide a theoretical basis for the prevention of Ebola virus disease, this survey aims to understand the distribution and prevalence status of Reston subtype of Ebola virus in north china area. Methods Collecting a variety of mammalian serums from zoos and farms in north china area, and detecting Ebola virus of Reston subtype. Results 109 kinds of collected mammalian serums, there were no positive samples detected by the Real-time PCR method. Conclusion Ebola virus has not been found in north china area, we should strengthen the publicity of Ebola virus disease, and do well security work.

Abstract:Objective To investigate the internal parasites infection in albino Marmota himalayan, and to provide a basis for seting up the quality standard of Marmota himalayana. Methods 21 wild albinism and 30 wild nomal Marmota himalayana from the same origin were detected by method as intestinal parasites in stool examination and serological testing, toxoplasma gondii and echinococcosis parasite infections of each animal, then detected the internal parasites and eggs under platoscope and microscope. Results The natural infectious rate of the roundworm in wild albinism and nomal Marmota Himalayana were respectively 71.4%(15/21) and 66.7%(20/30), using the exact probability method of inspection, there was no significant difference in the roundworm infection rate between two groups marmota; antibody test results of Serum toxoplasma gondii and Echinoocosis were negative. Application of albendazole and Ivermectin injection drug expelling and purification, effect to be trusted. Conclusions Monitoring results and the cure method can be used as an albino Himalayan marmot displacement experiment of animal parasite quality monitoring index and purification method.