Abstract:Objective To compare the extracellular space diffusion at different stages of rat C6-gliomas determined by MRI tracer method and analyze the influencing effect of extracellular matrix (ECM) on the diffusion process.Methods Introducing adolinium-diethylene triaminepentaacetic acid (Gd-DTPA) into extracellular space (ECS) as a tracer.The diffusion parameters and half-life time were quantified according to mathematical model of diffusion. The main ECM components (e.g. chordroitin sulfate proteoglycans (CSPGs),collagen IV tenascin C) were detected by immunohistochemical and immunoblot analysis. Results Gd-DTPA introduced into 20-day glioma in the rats diffused more slowly [(6.67±1.78) ×10^-5mm²/s vs. (1.26±0.27) ×10^-4mm²/s; t=4.265; P<0.01)], deriving a larger tortuosity [(3.99±0.57) vs. (2.83±0.29); t=4.11; P<0.01)], localized within the tumor with a smaller clearance rate [(7.67±2.29) ×10^-5mm²/s)vs.(1.46±0.36) ×10^-4mm²/s);t=3.87; P<0.05), and a longer half-life time((0.86±0.23 h)vs.(1.64±0.12 h); (t=5.91; p<0.01)] compared with 10-day gliomas in the rats. The increased levels of extracellular matrix of glioma were associated with different diffusion and clearance parameters of 20-day gliomas in the rats in comparision with those in the 10-day rat gliomas, in which the chordroitin sulfate proteoglycans[(0.48±0.07) vs.(0.32±0.09);t=4.663;P<0.01)], tenascin C [(0.29±0.04) vs. (0.58±0.11);t=6.50;P<0.01] and collagen IV [(0.24±0.07)vs.(0.33±0.06);t=3.81;P<0.05] were tested. Conclusions The ECS parameters are changed with the C6 glioma progression due to the increased ECM content. The results of our study may help us to better understanding the glioma micro-environment and provide beneficial references for the brain interstitial drug delivery to treat gliomas.

Abstract:Objective To observe the functional changes of T lymphocytes in the spleen of rats with bone cancer pain.Methods forty-one healthy female Sprague-Dawley rats were used in this study, and were divided into blank control, PBS and Walker-256 tumor groups. Bone cancer pain model was established by inoculation of Walker 256 cancer cells into the tibial cavity. The paw withdrawal threshold (PWT), paw withdrawal thermal latency (PWL), and spontaneous pain(SP) were all measured before modelling (as base) and at 4, 6, 8, 10, 12, 14, 16, 18, and 20 days after modelling.The function of T lymphocyte proliferation, and the content of T lymphocytes and their subgroups in the spleen were detected by cell counting kit-8 method and flow cytometry, respectively, on day 20 after modelling.Results Before modellng, there were no differences of PWT, PWL, and SP between the PBS and model groups. After modelling, the PWT and SP of model group were significantly decreased on day 4, and were always lower than that of PBS group during the experiment. Statistical analysis revealed that Walker-256 cancer cell inoculation in the tibia induced a significant decrease in PWL on day 8, 10 and 12 after modellng. Compared with the control group, T lymphocyte proliferation, content of T lymphocyte (CD3) and subgroups (CD4 and CD8) in the PBS group were not significantly decreased. However, T lymphocyte proliferation and the content of CD3 lymphocytes in the model group were significantly lower than those in the blank control group and/or PBS group.Conclusions The bone cancer pain rat model may appear obvious mechanical allodynia and spontaneous pain. Its thermal pain hyperalgesiaonly occurred in the intermediate stage of bone cancer pain. The content of T lymphocytes and its subgroups, and the function of T lymphocyte proliferation are weakened to some extent in the bone cancer pain rat model.

Abstract:Objective To study the real-time gait behavioral changes in rat models of experimental diabetic peripheral neuropathy.Methods Twenty-five SPF Sprague-Dawley rats were randomly divided into experimental group (6 males and 7 females) and control group (6 males and 6 females). Diabetes was induced in rats by intraperitoneal injection of streptozotocin (STZ) in a dose of 45 mg/kg. The gait behavior in all rats was tested at 12 weeks after diabetes modelling.Results Compared with the control group,the rats with diabetic peripheral neuropathy showed statistically significant different walk cycle extension, walk speed, average print intensity, balance and coordination. The abnormal gait behavior of the rat models was mainly reflected in the increased average and each foot walk cycle extension(P<0.01),average intensity (P<0.05), absolute average body rotation (P<0.01),and shortened both homologous coupling and homolateral coupling(P<0.05).Conclusions Experimental rat models of diabetic peripheral neuropathy can exhibit obvious changes of gait behavior, and may provide a reference for related clinical and basic research.

Abstract:Objective To define the loci of the mutant gene in the loop-tail mouse.Methods To study the heredity pattern, loop-tail mice were mated with normal C57BL/6J and C3H mice. Their offsprings with loop-tail or normal phenotype were registered respectively. Microsatellite marker D1Mit113 and D1Mit149 were used to locate the mutant gene. Based on fine mapping, the candidate gene Vangl2 was found. Vangl2 gene from the loop-tail mice was amplified by PCR followed by sequencing. Incision enzyme FspBI(BfaI)identified the genotype of offspring from loop-tail mice intercrossing. Results Heredity test indicated that the loop-tail phenotype was controlled by a single dominant gene not with 100% penetrance but was affected by genetic background. A C-to-T transversion was at the 1345bp in Vangl2 gene of the loop-tail mice.Conclusions The C-to-T transversion introduces a pre-termination codon of amino acids and causes the phenotype of loop-tail phenotype. None homozygous mice were found in the offsprings, suggesting that the homozygous mice are lethal.

Abstract:Objective Mongolian gerbil can make themself urine concentration for saving water and adapt to the harsh desert environment, due to their very unique moisture control system in the body. Methods Mongolian gerbil is resistant to drought on account of their special kidney. Histology of the kidney, ureter and bladder in Meriones Unguiculataus were observed by light microscopy using HE staining. Results The results showed that compared with rats and mice, the Mongolian gerbils have more developed distal tubules, and well developed inner renal medulla. Conclusions We hope that the findings of this study enrich our understanding of the histology of urinary system in Mongolian gerbils and provide support for the laboratory animalization of this animal.

Abstract:Objective To analyze and evaluate the population genetic quality of outbred KM mice from National Rodent Seed Center (Shanghai). Methods A total of 30 outbred KM mice were randomly chosen. The genetic characteristics of the population were determined by PCR and STR scanning using 30 selected microsatellite loci. Popgen1.32 software was used to process the data.Results Thirty microsatellite loci shared 123 alleles in the KM mouse population. The average effective allele number and the average heterozygosity were 2.3989 and 0.5342, respectively. The average polymorphism information content (PIC) was 0.4735. Conclusions The outbred KM mouse population of Shanghai Seed Center has genetic stability and genetic diversity, and is satisfied with the genetic characteristics of closed colony laboratory animal.

Abstract:Objective To analyze the population genetics of two outbred colony guinea pigs from two institutions and provide basical information in developing genetic detection methods and standardization of the outbred guinea pigs. Methods 25 polymorphic microsatellite markers were screened by a fluorescent based semi-automated genotyping method for the two populations of guinea pigs, and the population genetic parameters were calculated. Results A total of 121 alleles were detected in the two populations, with 2-10 alleles and a mean 4.84 alleles at each locus. The mean expected heterozygosity was 0.6067, and the average polymorphism information content was 0.552. In the two populations, 103 and 116 alleles were detected, the mean expected heterozygosity was 0.5195 and 0.5838, and mean polymorphism information content was 0.459 and 0.518, respectively. Five loci and six loci, respectively, showed significant deviation from Hardy-Weinberg equilibrium (P<0.05) in the two populations, mostly resulted from heterozygote deficiency. The average Fst of all loci was 0.1056, which implied a moderate genetic differentiation between populations. The Nei' (1972) genetic distance and Nei'(1978) unbiased genetic distance between the two populations were 0.3302 and 0.3204, respectively. Conclusions Both the two populations are consistent with a closed group of animal population genetic characteristics. Several loci deviate from HWE, which probably indicates that a certain degree of inbreeding phenomenon exists during the breeding process.

Abstract:Objective To improve the method of paraffin section preparation of whole mouse embryos and newborn mice. Method Specimens of mouse embryos and newborn mice were collected. Embryos aged 14.5, 15.5 and 16.5 days were directly placed in 4% neutral formaldehyde and fixed for more than 24 hours. For embryos aged 17.5 and 18.5 days, and 0-, 1- and 2-day old newborn mice, a small amount of formaldehyde was slowly injected into the chest, abdomen and brain at first, respectively, and then the whole specimens were fixed for more than 24 hours. Tissues were processed together using a fully-enclosed tissue processor. Paraffin-embedded slices were observed under microscope after HE staining. Results The quality of specimens following the improved method was better than before. The structure of different tissues from both mouse embryos and newborn mice can be easily determined. No fragmentation was found in any organ tissue under the light microscope. Conclusions We can handle whole body specimens of both mouse embryos and newborn mice simultaneously by adjusting the program of the fully-enclosed tissue processor. The staining results of the sections which were dehydrated using this improved method are clear and stable. This improved method may provide a useful approach in research of genetics and developmental biology.

Abstract:Objective To establish and apply an effective PCR assay for detection of the Tupaia (tree shrew) adenovirus (TAV).Methods According to NCBI Genbank, TAV genome DNA from 19418 to 19917 were synthetized and inserted into a plasmid as positive standards. One pair of primers was designed based on this sequence. Sixty blood samples and fifty-six stool samples from tree shrew were detected with this PCR assay.Results A PCR method for detection of TAV was successfully established, with a high specificity and the sensitivity was 13.5 ×10^-7 μg/mL. The PCR results of testing sixty tree shrew blood DNA samples were negative. 24 positive cases were tested among 56 stool DNA samples. Sequencing of the samples confirmed a 42.9% infection rate of TAV in tree shrew stool samples, well consistent with the PCR results. Conclusions The PCR method for detecting TAV established in this study has good specificity and high sensitivity, therefore, can be used in conventional detection of tree shrew adenovirus.

Abstract:Objective To establish a real-time fluorescent quantitative PCR (Q-PCR) method for detection of feline herpesvirus 1(FHV-1)in experiment cats and clinical sick cats. Methods Primers and TaqMan probes were designed and synthesized according to the published FHV-1 specific sequences of TK gene. FHV DNA standards were prepared using molecular biological techniques. The linearity, specificity, sensitivity, stability of the established Q-PCR method were tested. The method was used to detect 48 samples of cats. Results The linear range was 10² copies/μL to 10⁹ copies/μL. The developed Q-PCR method showed no cross reaction with herpes virus type 1 (HSV-1), canine herpesvirus (CHV), pig pseudo rabies virus (PRV) and cat parvovirus (FPV). The sensitivity was 10 copies/μL. The coefficient of variation (CV) was less than 5%. There were 33 positive cases detected in the 48 samples of cats. Conclusions The developed Q-PCR method is good in linearity, specificity, sensitivity, stability, and may be used for rapid quantitative detection of FHV-1 in cats.

Abstract:Phospholipase A2(PLA2)is a class of enzymes with the ability to catalyze hydrolysis of lipoproteins and glycerophospholipid in the membrane, mainly including secreted PLA2(sPLA2),cytosolic PLA2(cPLA2), Ca²⁺-independent PLA2(iPLA2) and lipoprotein-associated phospholipase A2(Lp-PLA2). They are closely related to many diseases.The studies of the role of PLA2 in cardiovascular diseases have been a hot topic in recent years. This paper focuses on the recent advances in research on the effects of PLA2 on cardiovascular diseases in animal experiments.

Abstract:Objective This review was aimed to provide reference for production,management and use of laboratory animals by analyzing the test results on intestinal parasitic infections of mice and rats in different provinces from 1989 to 2013 in China. The results showed that the infection rates in clean and SPF mice and rats were reduced to 10%,being better than that in the past years,but the situation was still not optimistic for the control of flagellate parasites infections.

Abstract:In order to study of etiology, pathogenesis, and therapy of endometriosis(EM) through animal models, we review the literature on rodent and primate models of endometriosis, including establishment of models and their applications. We hope that animal models provide useful tools for the studies of endometriosis.

Abstract:Vitrification provides a rapid cooling to induce glass-like solidification inside cells and protect cell membrane and cytoskeleton system free of injury by ice crystals. The main factors that can influence the effect of vitrification are the size of ovarian tissue, the kind of cryoprotectants, the method of permeation, carrier system and so on. Ovarian tissue structure is very complex, and there is no uniform standardized protocol of vitrification yet. This paper presents also the current problems of ovarian tissue vitrification.

Abstract:Human amniotic epithelial cells (HAECs) are isolated and extracted from placenta inner membrane and express embryonic stem cell surface markers.The HAECs are formed at 8th day from the epiblast after fertilization and possess multiple differentiation potentials. HAECs have the potential differentiation ability into all three germ layers: endoderm, mesoderm, and ectoderm. HAECs promote therapeutic and preventive effects and especially play an important role in treatment of central nervous system diseases. These stem cells inhibit inflammatory reaction and release neurotrophic factors to support the function of neurons. HACEs also exhibit alleviative effect on animal models of Alzheimer's disease (AD), Parkinson's disease (PD), spinal cord injury (SCI), stroke, etc. The present review summarizes the recent advances of therapeutic approaches of hAECs on central nervous system diseases and explores their protective mechanisms.