Abstract:Objective To study the function of Fkbp51 in the heart and liver by analyzing the differential RNA expression profiles in the wild-type mice (WT) and Fkbp51 knockout (KO) mice, and to elucidate the role of Fkbp51 gene in metabolic pathways in the heart and liver. Methods Using the second generation of high-throughput gene sequencing technology, the mRNA expression profiles of heart and liver were sequenced in WT and Fkbp51 KO mice. The data of sequencing of heart tissues were analyzed by DEGseq, and the results of sequencing of liver tissues were analyzed by BRB-Array Tools. The differential genes of the heart and liver in the mice were screened respectively. Gene ontology (GO) analysis and KEGG pathway analysis were performed to analyze the differentially expressed genes using the online tool DAVID. In addition, the differential genes of the two organ tissues were analyzed by Venn diagram. The interaction network of proteins was analyzed using the STRING database. Results (1) The absence of Fkbp51 led to changes in mRNA expressions of heart-related signal pathways such as vascular smooth muscle contraction, chemokine, retinol, and MAPK signaling pathways. (2) The lack of Fkbp51 mostly induced changes in cholesterol synthesis and metabolism, lipid metabolism, redox and other related genes and pathways in the liver. (3) In the heart and liver, Fkbp51 deletionresult ed in four co-differential genes, among them, down-regulation of Rnaset2b, Hmga1 and Fkbp51, while Cyp2b10 was down-regulated in the heart but up-regulated in the liver. All these proteins may interact with HSP90 protein and participat in the metabolism of heart and liver tissues. Conclusions Fkbp51 is involved in different metabolic and gene expression regulation pathways of heart and liver, and the roles are both independent and interrelated.

Abstract:Objective To establish a mouse model of H. pylori infection, and to evaluate the chronic pathological changes in the gastric mucosa associated with H. pylori infection. Methods 34 male 5~6-week old SPF C57BL/6 mice were used in this study. The mice were intragastrically administrated with a suspension of H. pylori SS1 strain. Two weeks after infection, rapid urease test and PCR were performed to confirm the H. pylori infection. Successfully infected mice were randomly divided into 3 groups including the control group, 6-week and 12-week infected groups. Samples of gastric mucosa were taken for pathological analysis using HE and borax methylene blue staining. The contents of myeloperoxidase (MPO), superoxide dismutase (SOD), malondialdehyde (MDA) and catalase (CAT) in the gastric tissues were detected by biochemistry, and the expression levels of COX-2, iNOS, TNF-α and IL-1β were examined by RT-qPCR. Results Compared with the control group, H. pylori colonization was observed in the gastric mucosa of the 6-week and 12-week infected mice, with chronic inflammatory cell infiltration, glandular atrophy and intestinal metaplasia to varying extents. The contents of CAT and SOD were significantly decreased, while the levels of MPO and malonaldehyde MDA, and the expression levels of COX-2,iNOS,TNF-α and IL-1β were significantly increased (P<0.05 or P<0.01). Conclusions Intragastric administration with H. pylori in C57BL/6 mice can be successfully used to generate the bacterial colonization, leading to chronic inflammatory cell infiltration, enhanced oxidative stress, and up-regulated expression of proinflammatory genes in the gastric glandular tissues at 6 and 12 weeks after inoculation. However, the inflammatory changes are more extensive in the mice at 12 weeks after infection, with glandular atrophy and intestinal metaplasia.

Abstract:Objective To investigate the effect of hydrogen-rich saline on apoptosis in hippocampal neurons induced by cerebral ischemia-reperfusion in rats and PI3K/Akt/FoxO1 signaling pathway. Methods The rat model of focal cerebral ischemia-reperfusion was established by thread-occlusion of the middle cerebral artery in rats. SD rats were randomly divided into sham operation group (Sham group), ischemia-reperfusion group (I/R group) and hydrogen-rich saline treatment group (HRS group), 10 rats in each group. At 24 h after reperfusion, the serum levels of IL-6, TNF-a and IL-1β were detected by ELISA. Histological changes of the hippocampus were observed by pathology using HE staining. Apoptosis in brain tissues was observed by TUNEL staining. The expression changes of p-PI3K, Akt, caspase-3 and FoxO1 proteins were detected by Western blot assay. Results Compared with the sham group, pyramidal cells were arranged loosely in the I/R group and a large number of pyramidal cells were necrotized, and the amount of apoptotic hippocampal cells was increased. The levels of IL-1β, IL-6 and TNF-α were significantly increased (P< 0.05), as well as the expression of p-PI3K, Akt and caspase-3 in the brain tissue. However, the expression of FoxO1 protein was decreased. There were significant differences between the two groups (P< 0.05). Compared with the I/R group, the inflammatory factors were significantly decreased in the HRS group. The expressions of p-PI3K, Akt and caspase-3 were also significantly decreased, while the expression of FoxO1 protein was increased (P< 0.05). Conclusions Hydrogen-rich saline can reduce brain injury caused by ischemia-reperfusion, and its mechanism may be related to PI3K/Akt/FoxO1 signaling pathway.

Abstract:Objective This paper reports a modified method for the isolation of rat thoracic aortic vascular smooth muscle cells based on combined enzyme double digestion. Methods An enzyme mixture containing collagenase Ⅱ, soybean trypsin inhibitor and elastase was prepared and used to remove the aortic tunica intima and tunica adventitia, and then the tunica media was subjected to second digestion using the same enzyme mixture and to isolate vascular smooth muscle cells. Results The isolated VSMCs were cultured in vitro and the growing cells had an elongated spindle-shape, reached confluence within a week, and displaying a typical "hill-and-valley" pattern. After 1 week, the cells were passaged. Contractile smooth muscle markers(α-SMA, myosin-Ⅱ and β-tubulin) were highly expressed in the isolated cells. More than 90% of the cells significantly expressed alpha smooth muscle actin and myosin-Ⅱ. Conclusions Themethod established in this study has advantages of simple and easy to operate, and good reproducibility, with a high activity and purity of the separated cells. It can ensure to obtain a large amount of contraction-type vascular smooth muscle cells within a short time.

Abstract:Objective To establish a stable and reliable mouse model as an alternative to the traditional model of impaired glucose tolerance induced by calorie restriction and its effect on glucose homeostasis. Methods Forty 16-week-old SPF C57BL/6J mice (half male and half female) were randomly divided into four groups by sex and the way of feeding. The mice in the ad libitum (AL) group had free access to basic diet, while the mice in the intermittent fasting (IF) group had normal diet and fasting on alternate days, with free access to water on the fasting days. The changes of body weight and blood glucose concentration in each group were monitored, and intraperitoneal glucose tolerance test and insulin tolerance test in mice were performed before and after the 12-week IF treatment. Results At 12 weeks after IF treatment, the body weight and blood glucose concentration of mice did not show significant difference. After i.p. injection of glucose, the blood glucose concentration of IF mice was less increased than the AL group, and after the insulin injection, the blood glucose concentration was more decreased. Compared to the AL group, the areas under the curve of tolerance test in the IF group were significantly decreased (P< 0.05). Conclusions After IF treatment, the mice show an enhanced sensitivity to insulin and improved glucose tolerance. This establishment method of mouse model of intermittent fasting is easy and simple, therefore, can be used as an effective alternative to traditional calorie restriction model of impaired glucose tolerance.

Abstract:Objective Monkey B virus(BV), also known as Cercopithecine herpesvirus 1,is an important zoonotic pathogen. According to the national standard, antibodies are detected using BV as an antigen. However, the preparation of BV antigen is very stricted due to biosafety issues. Therefore, in this study, we used alternative antigens to detect the BV antibody by serological assay and verified their specifity and sensitivity. Methods A total of 135 blood samples from rhesus monkeys were tested by two ELISA method (BV and HVP2) and enzyme immunosorbent assay (EIA)method. The positive and suspicious samples were verified by immuno-fluorescence assay (IFA), Western blot and immunoblotting technique using HSV-1 gC1 purified glycoprotein as an antigen. Results The positive rates of HVP2-ELISA, BV-ELISA and HSV-1-EIA were 32.6%, 37.8% and 34.8%, respectively. Consistant result of the three detection method accounted for 91.1% (123/135), and the positive result were confirmed by IFA And WB.There were 12 suspicious samples,in which 33.3% (4/12) were verified to be positive. Conclusions Compared with BV antigen, the sensitivity and specificity of the alternative antigen HSV-1 are moe close than HVP2. Positive and suspicious samples should be verified by several method to avoid missed detection.

Abstract:Objective To explore the population reproductive regularity and reproductive performance of rhesus and cynomolgus monkeys in Kunming area, and to provide reference data for the construction of rhesus and cynomolgus monkey breeding base, reproductive biology research and biological resources protection. Methods The reproductive regularity and reproductive performance in a rhesus monkey population of 20 males and 130 females, and a cynomolgus monkey population of 120 males and 780 females in a large-scale experimental monkey breeding base in Kunming were observed for a whole year and statistically analyzed. Results There was a distinct seasonal variation in the reproduction of rhesus monkeys in Kunming area, while no obvious seasonal changes was observed in cynomolgus monkeys. The pregnancy rate, reproductive rate and neonatal survival rate of rhesus monkeys were 76.15%, 69.23% and 90.70%, respectively. The pregnancy rate, neonatal reproduction rate and survival rate of cynomolgus monkeys were 78.98%, 74.87% and 94.81%, respectively. The menstrual cycle and pregnancy period of rhesus monkeys were (28.80 ±2.33) d and (165.87 ±7.52) d, respectively. The menstrual cycle and pregnancy period of cynomolgus monkeys were (29.35 ±3.05) d and (157.93 ±5.42) d, respectively. The average birth weight and weaning weight of rhesus monkey pups were (425.00 ±100.50) g and (1491.67 ±172.35) g, and those of cynomolgus monkeys were (314.33 ±61.18) g and (1013.50 ±115.50) g, respectively. Conclusions The reproduction regularity of rhesus and cynomolgus monkeys in Kunming area are defined. The reproductive performance of rhesus and cynomolgus monkeys are reported in detail. It provides basic data for the breeding of rhesus and cynomolgus monkeys in Kunming area and the development of research of experimental monkey reproductive biology.

Abstract:Objective To study the effect of drinking water at different pH on intestinal microbial diversity in SPF mice. Methods Ninety 21-day-old SPF mice were randomly divided into three groups:drinking acidified water (pH 3.0), neutral water (pH 7.0) and alkaline water (pH 9.0), respectively, for 3 months, 30 mice in each group. 10 mice were sacrificed at the end of 4, 8 and 12 weeks in each group. Bacteria, virus and parasites in serum were detected according to the national standard for SPF. Then, primer sequences were designed according to the conservativeness of bacteria 16S rRNA, and libraries were constructed. Subsequently, the ileocecal contents were subjected to high-throughput sequencing (Illumina). 10 G data were obtained and analyzed. Results The sequencing results revealed that the intestinal microbial abundance of the mice receiving pH 3.0 drinking water was moderate with a high stability. Conclusions Under the conditions of large-scale breeding, the mice receiving pH 3.0 drinking water can maintain the intestinal microbial diversity for a long time, and reach a stable platform. This study provides theoretical and technical support for the stable long-term maintenance of animal quality production. It is also a specific application of gastrointestinal macrogenomics in animal quality control.

Abstract:Objective To understand and compare the proportion of neural stem cells (NSCs) in the whole brain and cerebral cortex of mice at different embryonic days, and provide quantitative data for the later optimization of NSCs isolation and culture. Methods The whole brains (at embryonic 12.5, 14, 16 and 18 days) and cerebral cortex (at embryonic14, 16 and 18 days) were isolated and digested into single cell suspension, and were adherently cultured for 3-4 h.Immunofluorescence staining of Nestin, a NSCs specific marker, was used to statistically analyze the proportion of NSCs in each group.Expression of Nestin mRNA in the cerebral cortex of mice at E12.5, E14, E16, and E18 was detected by real-time fluorescence quantitative PCR. Results The result of immunofluorescence assay showed that there were Nestin-positive cells in the whole brain and cerebral cortex of mice at different embryonic days. In the whole brain,the proportion of NSCs was highest at E12.5 (53.42±1.57%) and lowest at E18(25.96±1.31%), and the proportions at E14 and E16 were placed in the middle among the groups. In the cerebral cortex, the highest proportion of NSCs was at E14 (33.65±0.29%), and the lowest at E18(25.29±0.28%), and the middle at E16 (26.82±0.30%).The result of real-time PCR showed that when the mRNA expression of Nestin in the cerebral cortex was set to 1, the relative mRNA expression of Nestin was 0.83±0.04 at E14, 0.77±0.05 at E16, and 0.44 ±0.05 at E18. Thus, the mRNA expression level of Nestin in the mouse cerebral cortex was gradually decreasing with the increase of embryonic days. Conclusions During the brain development, the proportion of NSCs is gradually decreasing in the whole brain and cerebral cortex of mice with the increase of embryonic days.

Abstract:Objective To observe the effects of road transport on hematological and biochemical parameters in New Zealand rabbits. Methods A total of 12 healthy New Zealand rabbits were selected for 2 h road transport. Blood samples were collected at 0, 24, 48, 72 and 96 h after transport, respectively. White blood cells (WBC), red blood cells (RBC), hemoglobin (HGB), hematocrit (MCV), mean erythrocyte hemoglobin content (MCH), mean erythrocyte hemoglobin concentration (MCHC) and platelets (PLT) were measured using a blood analyzer. Blood alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (ALB), total protein (TP), urea nitrogen (UREA), creatinine, uric acid (UA), triglycerides (TG), total cholesterol (COHL), glucose (GLU), hypersensitive C-reactive protein (CRP), α-amylase (AMYL), and creatine kinase (CK) were detected by an automatic biochemical analyzer. Results Compared the parameters before and after transport, The WBC count was increased first (P< 0.05 or P < 0.01) and then decreased after transport, the levels of RBC, HGB, HCT and PLT were decreased first (P< 0.05 or P < 0.01) and then increased after transport, and MCV was significantly high at 96 h after transport (P< 0.05). Among the clinical biochemical parameters, ALT, AST and BUN were firstly elevated (P< 0.05 or P < 0.01) and then decreased. TP, ALB as well as CREA and TG were firstly decreased (P< 0.05 or P < 0.01) and then increased. GLU was significantly low at 24 h after transport (P< 0.05). All parameters except MCV at 96 h after transport were not significantly different from those before transport. Conclusions Changes of blood routine, liver and kidney function indexes, lipid metabolism indexes, glucose metabolism index and creatine kinase index are observed in the New Zealand rabbits after 2-hour road transportation, and all the indicators except MCV return to pre-transport levels within 96 h.

Abstract:Objective To screen the markers of chromosome 12, which is vacant in the Beijing local standard DB11/T828.3-2011 for genetic quality control of miniature pigs in Beijing, and to study the applicability of the local standard by comparison of two different methods for evaluation of the genetic quality of the same population. Methods According to the literature, we selected four pairs of microsatellite markers of chromosome 12 and studied the polymorphism through monitoring the genetic quality of two populations of China Agricultural University miniature pigs. We screened the highly polymorphic markers of chromosome 12, combined them with the microsatellite primers of the standard 18 pairs of chromosomes to establish the whole chromosome method. We compared and analyzed the applicability of the local standard DB11/T828.3-2011 through monitoring the genetic quality of the same population of miniature pigs with different method. The data were processed and analyzed using software Popgen32. Results All the screened four pairs of microsatellite markers of chromosome 12 were highly polymorphic (PIC>0.5). The local standard showed a chromosome coverage of 94.7%, stability of amplification of 96.0%, and certified that the China Agricultural University miniature pig Ⅲ was qualified, while the China Agricultural University miniature pig I was not qualified. When the markers of chromosome 12 were added, the whole chromosome method showed result of 100% chromosome coverage and 96.6% amplification stability, both of the two populations of pigs were certified as qualified. Conclusions The four screened markers of chromosome 12 are all highly polymorphic, and provide a support for supplement of the local standard DB11/T828.3-2011.

Abstract:Objective To establish a rat model of superior mesenteric vein thrombosis by vein ligation and to simulate the pathological process of the disease, and to provide the basis for studies of its pathogenesis and treatment. Methods Ninety-six SPF male SD rats were randomly divided into three groups:Group A (sham operation group), group B (strangulation group) and group C (simple group), 32 rats in each group. Rats in group A were only opened the abdominal cavity but not blocked the blood supply. The rats were sacrificed at 8, 24, 48 and 72 h after operation. The rats in groups B and C were subjected to establish the strangulation and simple models by superior mesenteric vein thrombosis, respectively, and were sacrificed at 8, 24, 48 and 72 h after modeling. Histological changes (H&E staining) in the rat intestinal tissues were evaluated by a pathological scoring system. The levels of intestinal fatty acid binding protein (IFABP) and α-glutathione S-transferase (α-GST) were detected by ELISA. Results The rat model of mesenteric vein thrombosis was successfully established, with a success rate of 100% (96/96). The pathological analysis revealed that compared with the group A, different degrees of blood stasis and injuries were observed in the intestinal tissues of groups B and C, and the injury were gradually increased in the group B, while gradually reduced in the group C. The degrees of blood stasis and injury were positively correlated with the scope of ligation. The result of ELISA showed that the serum levels of IFABP and α-GST of the rats in groups B and C were significantly higher than those in group A (P < 0.05), and the degree of elevation was positively correlated with the scope of ligation. Conclusions In this study, the rat model of superior mesenteric vein thrombosis is successfully established by vein ligation. This model is simple and easy to operate with a high success rate, and can be used in related research.

Abstract:Objective To observe the promoting effect of hypoxia on proliferation of human bone marrow mesenchymal stem cells (BMMSCs) and maintaining their potential of multi-directional differentiation in vitro. Methods BMMSCs were isolated from bone marrow blood samples of patients with bone fracture, and cultured under hypoxic (group A) or normoxic (group B) conditions. The morphology, proliferation and osteogenic and adipogenic potential of the BMMSCs were observed. Results BMMSCs in the group A showed a long spindle shape and a fish shoal-like distribution, and were well-grown, while the morphology of cells in the group B appeared polygonal or flat. The quantity and growth rate of BMMSCs in the group A were increased compared with the group B (P< 0.05), with an osteogenic and adipogenic potential. Conclusions Hypoxia can promote the proliferation of BMMSCs in vitro and maintain their multi-directional differentiation potential.

Abstract:Objective To investigate the expression and significance of Maspin and IKKα in nasosinusoidal mucosa of rats with fungal rhinosinusitis (FRS). Methods A total of 40 SD rats were used to establish the FRS model, and randomly divided into nasal obstruction group, FRS group, immunosuppressive group and invasive FRS group, 10 rats in each group. Another 10 normal rats were used as control group. Mice in the control group were fed with normal diet. In the nasal obstruction group, the mice had only hemostatic cotton stuffed in the nasal cavity and injection of 0.9% NaCl in the abdominal and nasal cavities. In the FRS group, the mice were injected Aspergillus fumigatus spore suspension into the nasal cavity and 0.9% NaCl i.p. The mice of the immunosuppressive group were given cyclophosphamide i.p. and 0.9% NaCl injection into the nasal cavity. The invasive FRS group was injected with cyclophosphamide i.p. and Aspergillus fumigatus spore suspension into the nasal cavity. The serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA). The expression of Maspin and IKKα in nasosinusoidal mucosa was detected by immunohistochemical staining. The expression of Maspin mRNA and IKKα mRNA in the nasosinusoidal mucosa was detected by fluorescence quantitative PCR. Results The serum levels of IL-6 and TNF-α in different groups were significantly different (P< 0.05). The level of IL-6 in the FRS group was (69.3 ±10.9) ng/L, significantly higher than those in the control group, nasal obstruction group, immunosuppressive group and invasive FRS group[(45.2 ±7.1)ng/L, (46.4 ±6.7) ng/L, (21.3 ±4.5) ng/L, and (20.9 ±4.3 ng/L)] (P < 0.05). The level of TNF-α in the FRS group was (30.4 ±4.8) ng/L, significantly higher than those in the control group, nasal obstruction group, immunosuppressive group and invasive FRS group[(14.8 ±2.7) ng/L, (13.9 ±1.4) ng/L, (7.9 ±0.6) ng/L, and (7.8 ±0.4 ng/L)] (P < 0.05). The levels of IL-6 and TNF-α in the control group were significantly higher than those in the immunosuppressive group and invasive FRS group (P< 0.05). There was no significant difference between the immunosuppressive group and the invasive group (P> 0.05). Theresult of immunohistochemical staining showed that the protein expression of Maspin in the FRS group and invasive FRS group was significantly lower than that in the control group, nasal obstruction group and immunosuppressive group, while the expression of IKKα protein was significantly higher than that of control group, nasal obstruction group and immunosuppressive group (P< 0.05). The protein expression of Maspin in the invasive FRS group was significantly lower than that in the FRS group, by contrast, the expression of IKKα protein was significantly higher (P< 0.05). The PCRresult revealed that the expression levels of Maspin and IKKα mRNA were (0.217 ±0.013) and (0.193 ±0.012), significantly lower than that in the control, obstruction and immunosuppressive groups [(0.309 ±0.021),(0.302 ±0.017),and (0.293 ±0.02)](P< 0.05), while the expressions level of IKKα mRNA were significantly higher [(0.319 ±0.043),(0.384 ±0.048) vs (0.169 ±0.015),(0.171 ±0.018),and (0.175 ±0.019)](P< 0.05). Conclusions Down-regulation of Maspin expression after IKKα activation is the main cause of the onset of FRS, which may also be one of the mechanisms of invasive FRS.

Abstract:Objective To detect the expression of long non-coding RNA (lncRNA) CCAT1 in human papillary thyroid cancer, and to observe the effect of CCAT1 down-regulation on the invasion and migration of human papillary thyroid cancer. Methods The expression of CCAT1 was detected in human normal thyroid Nthy-ori 3-1 cells and human papillary thyroid cancer TPC-1 cells. CCAT1 siRNA plasmid was transfected into TPC-1 cells. The effect of CCAT1 down-regulation on cell invasion and migration was observed by Transwell chamber assay and scratch test, and the expressions of BRAF, MUC15 and RKIP proteins were detected by Western blot. Results The level of CCAT1 in human papillary thyroid cancer TPC-1 cells was significantly higher than that in human normal thyroid Nthy-ori 3-1 cells. CCAT1 down-regulation significantly inhibited the invasion and migration of TPC-1 cells. The Transwell invasion assay revealed that the number of migrated TPC-1 cells in the CCAT1 down-regulation group was significantly lower than that in the control group. The scratch test showed an increased distance between cells in the CCAT1 down-regulation group compared to the control group, suggesting a reduced cell motility. The expressions of BRAF and MUC15 proteins were decreased in the CCAT1 down-regulation group, while that of RKIP protein was increased. Conclusions The expression of CCAT1 in papillary thyroid cancer cells is significantly higher than that in normal human thyroid cells. Down-regulation of CCAT1 in papillary thyroid cancer cells may inhibit the cell invasion and migration by regulating the expression of BRAF, MUC15 and RKIP proteins.

Abstract:Objective To compare and analyze the genetic structure of NIH mice bred in Unites A and B, using microsatellite technology. Methods Thirty SPF 8-week old outbred NIH mice (half male and half female) of each population were randomly chosen from the Units A and B, respectively. PCR amplification and STR scan were performed to determine the genetic characteristics of two outbred populations using microsatellite loci, and the population genetic structure was analyzed with statistical software Popgene 1.32. Results In the NIH mouse population form the Unit A, 74 alleles were obtained, with an average heterozygosity of 0.3108 and polymorphism information content of 0.2637. In the NIH mouse population from the Unit B, 76 alleles were obtained, with an average heterozygosity of 0.3257 and polymorphism information content of 0.2777. The inter-population comparison showed that genetic differentiation coefficient Fst was 0.3932, the genetic identity was 0.3971, and the genetic distance was 0.9235. The population difference was significant. Conclusions There is serious genetic differentiation between the two NIH mice populations, resulting in the formation of two different closed populations.

Abstract:Objective To explore the effects of cyclophosphamide administered by different routes or in different doses on the embryo-fetal development in pregnant rabbits, and to determine the optimal mode of cyclophosphamide administration to induce fetal malformation. Methods Pregnant rabbits were divided into control group C (saline), group Y1 (intravenous injection of 15 mg/kg cyclophosphamide,), group Y2 (subcutaneous injection of low dose cyclophosphamide, 20 mg/kg), and group Y3 (subcutaneous injection of high dose cyclophosphamide, 30 mg/kg). Each rat was administrated according to the corresponding mode once daily on GD10~13. The day of conception was designated as GD0. The pregnant rabbits were sacrificed and dissected on GD28. Then, the number of corpora lutea and implantation, the weight of uterus with contained fetus, and fetal resorption rate were detected, the fetuses were removed and the fetal sex, body length, tail length, the number of live births and stillbirths were recorded, and the appearance of deformities, visceral deformities and skeletal malformations were detected. Results Pregnant rabbit fetuses in the cyclophosphamide intravenous injection group and subcutaneous injection of low dose cyclophosphamide group showed deformities. The appearance malformation rates in the two groups were 30.77% and 95.65%, the skeletal deformity rates were 7.69% and 73.91%, and the visceral abnormality rates were 20.51% and 47.83%, respectively. The fetal resorption rate in the high dose cyclophosphamide subcutaneous injection group was 100%. Conclusions Subcutaneous injection of 20 mg/kg cyclophosphamide to pregnant rabbits at GD10~13 can be used as a positive administrationmethod for rabbit embryo-fetal developmental toxicity test. Thismethod has the advantages of short administration period, easy operation, few fetus resorption, and high rate of fetal malformation, thus, providing the evidence for selection of appropriate model of rabbit embryo-fetal developmental toxicity.

Abstract:The method for preparation of models of myocardial ischemia in rats have been well described in the literature, are of practical value and have been chosen by many researchers for pharmacological studies of drugs for human diseases. However, there is still lack in some details of their operability and practicability. We re-selected the coronary artery ligation site, simplified the procedures and improved the experimental method for preparation of the models, and made satisfactory result. In this paper we will review the selection of method for preparing myocardial ischemia model in rats, describe some details of the surgical operation, explored the influencing factors and so on, and provide a reference for selecting most appropriate animal model in research.

Abstract:Infertility has become a global problem affecting human reproductive health. As an important treatment for infertility, assisted reproductive technology has made great progress over the past few decades. Rapid development has also taken place in medical devices for human assisted reproductive technology. It is imperative to establish the risk management and safety evaluation system of these products. In 2016, the industry standard YY/T 1434-2016 "Human in vitro Assisted Reproductive Technology With Medical Equipment in vitro Mouse Embryo Test" was officially released. In this paper, the key notes and elements of this in vitro mouse embryo test are briefly reviewed.

Abstract:A safe and effective anesthesia technique is necessary in ensuring a successful surgical operation in rabbit experiments. A variety of anesthesiamethod have been reported, yet, no one matured and widely accepted anesthesiamethod is available so far. This article aims to provide an information basis for further research on general anesthesia in rabbits by reviewing the literature on single and combined anesthesia techniques in rabbits reported in the last decade.