Abstract:Objective To investigate the distribution of histone deacetylase 2 (HDAC2) in the hippocampus of adult C57BL/6 mice and the co-localizations of HDAC2 and PSD components, such as N-methyl-D-aspartate (NMDA) receptor 1 (NR1) and PSD-95, and provide morphological evidence for inherent relationship between postsynaptic density (PSD) components in synapses and HDAC2 in nucleus. MethodsThe morphology and distribution of HDAC2 positive neurons in the hippocampus were observed by immunohistochemistry. The hippocampal distributions of NR1 and PSD-95, as well as co-location with HDAC2 were detected using double immunofluorescence staining. ResultsHDAC2 immunoreactivity was mainly observed in hippocampal CA1-CA3 pyramidal cells and dentate gyrus granule cells. The scattered positive cells were accidentally present in the oriens layer, radiate layer and lacunosum-moleculare layer of CA1-CA3 and molecular layer and polymorphic layer of dentate gyrus. The distributions of NR1 and PSD-95 positive neurons are similar to that of HDAC2 positive neurons in the hippocampus. The co-location analysis showed that almost all HDAC2 immunoreactive neurons observed in pyramidal cells of CA1-CA3 and granule cells of dentate gyrus. Conclusions HDAC2 is rich in hippocampal pyramidal cell layer and granule cell layer, and co-located with PSD proteins. The results provide a morphological clue to explain the role of HDAC2 in postsynaptic glutamatergic neuron-dependent synaptic plasticity. 

Abstract:Objective To investigate the effect of IL-21 on secretion of IFN-γ in CD8⁺ T cells of rhesus monkeys infected by SHIV. MethodsThe in vitro study was carried out on the culture of CD8⁺ T cells isolated from rhesus monkey models infected by SHIV. CD8⁺ T cells were incubated with various concentrations of IL-21 (1～15 ng/mL). Then IFN-γ concentrations in the supernatant were detected by ELISA, RT-PCR was performed to detect the expression of IFN-γ mRNA in the CD8⁺ T cells, and the percentage of IFN-γ-positive CD8⁺ T cells was assayed by flow cytometry. ResultsIL-21 (10 ng/mL) significantly increased IFN-γ level (P <0.05) in the supernatant of CD8⁺ T cell culture, compared with that in the control group. The expression of IFN-γ mRNA was significantly enhanced. IFN-γ which requires de novo synthesis following activation peaked at 4 h post stimulation. ConclusionIL-21 can increase the secretion of IFN-γ in vitro. 

Abstract:ObjectiveTo explore the relationship between DJ-1, DJ-1M26I, DJ-1L166P with the cell proliferation and apoptosis of NIH 3T3 cells at cellular level, and provide a basis for the construction of a transgenic animal model of Parkinson’s disease and further study on the pathogenesis of this disease. MethodsRecombinant plasmids pcDNA3.1/myc-His-DJ-1, pcDNA3.1/myc-His-DJ-1L166P and pcDNA3.1/myc-His-DJ-1M26I were transfected into NIH 3T3 cells, respectively, using lipofectamine. The cells were screened with G418 at a dose of 500 μg/mL. Stable clones were identified on the DNA, RNA and protein levels. MTT assay and annexin V-FITC kit were used to detect the viability and apoptosis of those stable cell clones. ResultsAfter the G418-screening of of NIH 3T3 cells transfected with recombinant plasmids pcDNA3.1/myc-His-DJ-1, pcDNA3.1/myc-His-DJ-1L166P or pcDNA3.1/myc-His-DJ-1M26I, one, four and three positive clones were obtained, respectively, by PCR detection. RT-PCR and Western blot detected the expression of DJ-1-His in the positive clones. NIH 3T3 cells transfected with DJ-1L166P and DJ-1M26I had a higher expression of caspase-3 mRNA than normal NIH 3T3 cells, while NIH3T3 cells transfected with DJ-1 had a lower expression. MTT assay showed that NIH 3T3 positive cells transfected with DJ-1L166P and DJ-1M26I had a lower proliferation rate than that of normal NIH3T3 cells (P<0.05), while the NIH 3T3 positive cells carrying DJ-1 gene did not show significant difference compared with the normal NIH 3T3 cells. Apoptosis test indicated that the apoptosis rates of DJ-1L166P and DJ-1M26I transfected cells were higher than that of normal NIH 3T3 cells, however the apoptosis rate of the DJ-1-transfected cells was significantly lower than that of normal NIH 3T3 cells (P<0.05). ConclusionsDJ-1L166P and DJ-1M26I mutations reduce the proliferation of NIH 3T3 cells. DJ-1L166P and DJ-1M26I mutations also enhance apoptosis in NIH 3T3 cells. Their effects on NIH 3T3 cell proliferation and apoptosis are similar.

Abstract:Objective To explore whether streptozotocin-induced diabetic rat models are suitable to evaluate the pharmacokinetic processes of glibenclamide in the diabetic state. MethodsDiabetic rats induced by intraperitoneal injection of 60 mg/kg streptozotocin and normal rats were administered glibenclamide at a dose of 10 mg/kg. The concentration of glibenclamide was determined by high performance liquid chromatography. Pharmacokinetic parameters were calculated using DAS 2.0 software. ResultsThe pharmacokinetic parameters of glibenclamide in normal rats were as following:T_max was 84.784 min, C_max was 0.259 mg/L, CL/F was 0.092 L/min/kg and AUC(0-720min) was 509.523 mg/L·min. The same parameters in diabetic rats:T_max was 255.427 min, C_max was 0.910 mg/L, CL was 0.092 L/min/kg and AUC(0-720min) 1528.280 was mg/L·min. ConclusionsThere were significant differences between pharmacokinetic values of normal and diabetic rats. The results were different from that reported in the literature. Therefore, streptozotocin-induced diabetic rat models might not be suitable for pharmacokinetic evaluation of drugs in type Ⅱ diabetic conditions.

Abstract:Objective To identify the P. mirabilis strain isolated from infant rhesus monkeys in the period of artificial feeding, and provide useful data for detection and identification of the bacteria in laboratory rhesus monkeys. MethodsP. mirabilis strains isolated from infant rhesus monkeys were identified by cultural characteristics, colony morphology, staining, motility and biochemical test and serological test. Meanwhile, antibiotic sensitivity, virulence and toxin tests were conducted.ResultsThe results of cultural characteristics, colony morphology, staining, biochemical and serological tests showed that the isolate was a strain of P. mirabilis. Effective therapeutic drugs were screened by antibiotic sensitivity test, and the disease was effectively controlled. Virulence tests showed that it is a highly pathogenic strain to mice. ConclusionsThe results of this study demonstrate that P. mirabilis isolated from infant monkeys is the pathogenic agent leading to their diarrhea and death. This bacteria strain is a conditional pathogen, which may cause some potential harm to researchers and laboratory rhesus monkeys. Although laboratory rhesus monkeys are allowed to be P. mirabilis carriers in our country, it is worth to pay high attention to the serious consequence of P. mirabilis infection, and it should be considered as an important issue. 

Abstract:Objective To investigate the relationship between tissue tropism of influenza viruses and sialic acid receptors in ferret. MethodsThe distribution of influenza viruses H5N1 (SZ406H, A/VN/1203/04), SH1N1, H3N2(Brisbane/09, HK/09) in infected ferrets was analyzed by virus isolation, and the distribution of sialic acid receptors in tissues was determined by direct immunofluorescence assay. The binding of influenza virus to sialic acid receptor was detected by indirect immunofluorescence assay. ResultsH5N1 (SZ) and H5N1 (A/VN/1203/04) viruses were detected in the liver, spleen, lung and intestine. In addition, H5N1(A/VN/1203/04) virus was also detected in the brain. SH1N1 and H3N2(Brisbane/09, HK/09) viruses were detected in the intestine. Sialic acid receptors type I SAα2,6Gal and SAα2,3Gal were distributed in the spleen, heart, lung, intestine, and brain, type II SAα2,3Gal receptors were distributed in the spleen, heart, lung, intestine, brain, and liver. SH1N1 virus was bound to SAα2,6Gal in the tissues, while H5N1 was bound to SAα2,3Gal. ConclusionsOur results indicate that H5N1(SZ,VN) virus can be distributed in many organs in ferret. SH1N1 and H3N2(HK,BR) viruses are only distributed in the intestine, while both SAα2,3Gal and SAα2,6 Gal receptors are distributed in many organs. So that we would conclude that SA receptors are not determinant factors for organ distribution of different subtype influenza viruses in ferrets. 

Abstract:To understand the genetic characteristics of Shanyi colony inbred strain E of Chinese hamster, the reproduction records of Shanyi colony inbred strain E of Chinese hamster in 2001-2010 were used to estimate the heritability of the reproduction traits by the method of average information restricted maximum likelihood (AIREML). The results showed that the heritability of the litter size, weaning number, birth interval 2-1, birth interval 3-2 was low, with the value of 0.5,0.6,0.182 and 0.116, respectively. 

Abstract:Objective To analyze the genetic diversity and phylogenetic of Macaca mulatta in Yunnan Province, which provide important reference and background data for researches of macaca mulatta protection and reasonable application. MethodsNinety six blood samples of Macaca mulatta were collected for amplifying the complete sequences of mitochondrial DNA control region by polymerase chain reaction (PCR). The variation sites, parsimony informative sites, haplotypes, haplotype diversity, and nucleotide diversity were analyzed using Mega 4.0 and DNA SP software. The phylogenetic tree was constructed by neighbor-joining (NJ) and minimum-evolution(ME) methods.ResultsOne hundred and forty-nine variation sites and 46 haplotypes were found in the ninety-sixth Macaca mulatta, the haplotype diversity (Hd) was 0.968±0.007 and nucleotide diversity (Pi) was 0.020. The phylogenetic analysis based on mitochondrial control region showed that these Macaca mulatta lived in Yunnan was divided into two sister groups geographically. ConclusionsThe results of this study indicate a high variation and phylogenetic evolution relationship of Macaca mulatta in Yunnan province. 

Abstract:Objective To design and manufacture a practical experimental feed for tree shrew breeding and growth, according to the biological characteristics of tree shrew and relevant standards for laboratory animal feed. The results indicate this feed can meet the needs of growth, reproduction and experiments of tree shrews. Both the feed nutritional ingredients and uniformity of admixture are accord with local governmental standards for tree shrews. It is not only convenient to manufacture and feed but also promising in application and popularyzation. 

Abstract:ObjectiveTo establish the normal range of body weight and blood biochemical indexes of Tianjin wild (TW) inbred mice, and to determine their coat color gene. MethodsThe body weight and blood biochemical parameters of TW inbred mice were examined. The coat color gene was determined by mating with DBA/2 mice. ResultsWhen the TW mice were younger than six weeks old, there was no significant difference between the body weights of TW male and female mice (P<0.05). When the TW mice were older than six weeks of age, the body weight of TW male mice was higher than that of TW female mice (P<0.05). Compared with the female group, the total protein was higher in male group (P<0.05), while the triglycerides were lower (P<0.05). The coat color gene analysis revealed that hair color of first generation hybrid was white belly wild color, and its gene type was AWAWBBccDD. ConclusionsTW inbred mice have their own characteristic blood biochemical indexes, and some are different from those of commonly used inbred mouse strains. The coat color gene of TW mice is homozygosis. 

Abstract:ObjectiveThe targeting fusion protein XE-TNFαm2 produced by the genetic engineered E. coli strain DH5α/pCW-PL-XE-TNFαm2 was shown a prospect to effectively eliminate the HIV in AIDS patients. Its expression level reached 32%~36% of total cell soluble proteins. The purpose of this study was to investigate the hereditary stability of this engineered strain. MethodsThis strain was transferred on LBAmp⁺ and LBAmp- solid culture media day by day as generation by generation for a total of 100 generations, and then stored at 4℃ for 4,5, 6 months, respectively. Finally, to check the protein XE-TNFαm2 (20.3 kDa) expression levels of each transferred generation strain on LBAmp⁺ and LBAmp- media cases, respectively, by general protocol for temperature control expression, so as to compare the expression levels in the above different cases. The samples were taken from generations transferred for 1,0, 0,0, 0,0, 0,0, 0,0, 100 days on LBamp⁺ and LBAmp-media, respectively. ResultsThe protein XE-TNFαm2 expression levels were not obviously changed from the 1st generation to 100th generation under LBamp⁺ and LBAmp- cases, respectively (P>0.05). Only in the case that after the strains were transferred for 100 generations on LBamp⁺ or LBAmp-media and then stored at 4℃ for 4,5 or 6 months, respectively, the protein XE-TNFαm2 expression levels were reduced by 8%. ConclusionsCIts857 sequence, PL promoter and termination T1T2 sequence, existing in this plasmid vector, are three key elements to maintain such a high expression level. This genetic engineering E. coli strain DH5α/pCW-PL-XE-TNFαm2 has good hereditary stability. 

Abstract:ObjectiveTo explore the effect of an AT1 inhibitor losartan on the expression of TNF-α and myeloperoxidase (MPO) in the lung of mechanically ventilated rats, and further study the protective effect of losartan on ventilator induced lung injury. MethodsTwenty-four healthy male Wister rats were randomly divided into three groups:control group, ventilator induced lung injury (VILI) group and losartan group. ELISA in situ was used to determine the blood content of angiotensin II (Ang Ⅱ) in the lung tissue, immunohistochemical staining was employed to detect the expression level of TNF-α protein, and colorimetry was applied to measure the activity of myeloperoxidase (MPO) in the homogenate of lung tissue. ResultsThe Ang Ⅱ concentration and expression level of TNF-α protein in the lung tissue, and MPO activity in the homogenate of lung tissue of the VILI group were significantly higher than those in the control group (P<0.01). There was no significant difference between the Ang Ⅱ concentration and MPO activity in the losartan group and VILI group (P>0.05). However, the expression level of TNF-α protein, MPO activity and wet/dry lung weight (W/D) ratio were significantly decreased in the losartan group in comparison with that of the VILI group (P<0.01), and the lung injury was apparently less serious than that in the VILI group. ConclusionsAng Ⅱ regulates the expression of TNF-α in the lung tissue and play an important role in the pathogenesis of ventilator-induced lung injury. Losartan may block the binding of Ang Ⅱ and AT1 receptor, down-regulate the expression of TNF-α protein and decrease the MPO activity of lung tissue homogenate, and may have preventive and therapeutic effect on ventilator-induced lung injury to some degree.

Abstract:ObjectiveTo establish a GK rat model of type 2 diabetic nephropathy induced by high-calorie and high-protein diet, and to explore its possible mechanism of action. MethodsA total of 24 28-week old GK rats were randomly divided into normal group and model group, 12 in each group. The normal rats were given normal diet while the model rats were fed a high-calorie and high-protein diet for 8 weeks. 24 h-U-ALB, 24 h-U-TP, Ucr, U-ALB/Ucr were determined at week 0,4, and 8. FBG, Scr, BUN, TC, TG, NO were measured at week 4 and 8. The rats were sacrificed 8 weeks later, and the two kidneys of each rat were taken and weighed to calculate the kidney hypertrophy index. The renal pathological changes were examined and the Na⁺K⁺-ATPase activity in renal tissue was detected. ResultsThe 24 h-U-ALB, 24 h-U-TP, U-ALB/Ucr, FBG, TC, TG, NO, kidney hypertrophy index and Na⁺K⁺-ATPase activity were significantly increased in the model group rats. They showed an increased glomerular volume of the kidney, mesangial matrix hyperplasia and thickening of the glomerular basement membrane. ConclusionsA GK rat model of type 2 diabetic nephropathy is successfully established by a high-calorie and high-protein diet. The rise in blood glucose and lipid levels contribute to the development of diabetic nephropathy. Besides, increased activity of Na⁺K⁺-ATPase further damages the function of renal tubules. The increased NO leads to glomerular hyperperfusion and hyperfiltration. These factors also accelerate the formation of nephropathy in GK rats. 

Abstract:ObjectiveTo compare the biological characteristics of three modeling methods to establish human large cell lung cancer (LCLC) NCI-H460 xenograft models in nude mice, and provide an experimental basis for finding a suitable modeling approach for different studies. MethodsCultured NCI-H460 cells, NCI-H460 tumor tissue pieces and tumor homogenate were used respectively to establish subcutaneous tumor models in BALB/c-nu/nu nude mice. The tumor formation rate, tumor weight, doubling time and morphology were observed. Blood alanine aminotransferase (ALT), aspartate aminotransferase (AST), glucose (G1u), urea nitrogen (BUN) and creatinine (CREA) were analyzed using an automatic biochemical analyzer. WBC count and classification were determined, and finally the phagocytic activity of macrophages and NK cell activity in vitro were assessed. ResultsThe tumor formation rate, tumor growth rate and uniformity in the cultured cells and tumor homogenate groups were significantly better than those of the tissue transplantation group. Five weeks after inoculation, the blood ALT and AST were significantly increased, while BUN, CREA were significantly lower in all the three groups of tumor-bearing mice, the AST and BUN of the tissue transplantation group were significantly higher than those of the other two groups. However, the Glu of tumor-bearing mice decreased significantly 2 weeks after inoculation. The LYM%, MN% and HGB of the peripheral blood of the three groups were lower than those in the normal control group, and WBC, NEUT%, PLT of the cultured cell and homogenate groups were significantly higher than those of the tissue transplantation group. The immune cell activity of the macrophages and NK cells showed a lowering trend in an order of normal control group>homogenate group>cultured cell group>tissue transplantation group. ConclusionsThe modeling method with cultured cells is controllable in number and uniform of tumor growth. It is suitable for the establishment of different experimental xenograft models. The tissue transplantation can meet the need for the establishment of animal models to serve Chinese anti-cancer drug screening, and the homogenate inoculation method is not recommended. The physiological and immunological characteristics of the mouse models have a close relationship with tumor growth.

Abstract:ObjectiveTo establish a rat model of transplanted lung cancer by intravenous injection of tumor cells and evaluate the feasibility of this method. MethodsThree doses of Walker-256 tumor cells were intravenously injected into the tail vein in rats:the high dose group (1.5×10⁶cells/mL), mid-dose group (0.5×10⁵ cells/mL), and low dose group (2.5×10⁴ cells /mL), 10 rats in each group. The survival time, body weight, tumor formation rate in the lung and metastasis to other organs in the rats of different groups were observed. The lung and several other organ tissues were examined by histopathology. ResultsCompared with the normal group, the body weight and daily food intake of the rats in model groups were markedly decreased 14 days after the injection of tumor cells. Some rats of the high-dose group began to die at 21 days after the injection of tumor cells. Therefore, at the 21 days after the injection of Walker tumor cells, all rats were sacrificed and organ samples were taken for pathological examination. The tumor formation rate in the rat lungs of the high-dose,mid-dose and low-dose groups were 100%, 80% and 30%, respectively. The liver coefficient and lung coefficient of model groups were much higher than that of the normal group. Pathological examination revealed that tumor nodules occurred in the rat lung tissues of model groups, but no was found in the liver, spleen and other organs. ConclusionsIntravenous injection of high-dose of Walker-256 cells can be used to establish successfully transplanted lung cancer model in rats, therefore, to provide a useful method for establishment of rat models of lung cancer and serve further experimental studies of lung cancer.

Abstract:ObjectiveTo establish a loop-mediated isothermal amplification (LAMP) technique for detecting Staphylococcus aureus in laboratory animals. MethodsFour primers were designed and screened specifically to recognize the nuc gene of Staphylococcus aureus. Optimal reaction conditions were screened. Results2 cfu extracted crude DNA and no cross reaction with other commonly seen pathogenic bacteria was found when the conditions were 60℃ for 40 min at least, Mg²⁺at a concentration of 8.0 mmol/L, dNTP 2.0 mmol/L and betaine 0.8mmol/L. The result of reaction can be directly observed with naked eye by adding a fluorescent SYBR green I dye. ConclusionsThis LAMP technique for detectiong Staphylococcus aureus is sensitive, rapid and simple to perform, and can satisfy the requirements of grassroots lab, mass samples and quick detection.

Abstract:Animal models of liver cancer are an important platform and method for research of etiopathogenesis and mechanism of liver cancer, and it is important to establish animal models of liver cancer to serve liver cancer research．Establishing better animal models of liver cancer can provide experimental and theoretical references for studies of pathogenesis, diagnosis and treatment of liver cancer．

Abstract:The biochip technology is used to build micro-biochemical analysis systems on the chip surface, through micro-processing and micro-electronics technology. It can make high-throughput detection of tissue cells, proteins, DNA or other biological components. Biochips are widely used for life sciences, judicial expertise, food and nutritional science, environmental science, agriculture and forestry science, military science and other areas. This article makes a brief summary of the application of biochip in cancer research, diagnosis and treatment.

Abstract:Along with the socio-economic development, acceleration of urbanization process and continuous improvement of living standard,more and more pets are bred in urban households and playing an important role in people''s everyday life．However,many pets are important carriers of infectious zoonotic diseases, especially dogs and cats．In more than 200 kinds of major reported zoonoses, there are over 70 kinds directly or indirectly related to dogs and cats. With the rapid development of pet industry and increasing number of dogs and cats, the incidence of pet-derived zoonoses are increasing, which cause some problems to their prevention and control. In order to improve the pet management system, establish an efficient epidemic prevention and supervision system, and effectively prevent and control the zoonoses in dogs and cats, strategies are discussed in this article, including perfecting the laws and regulations, forming a comprehensive management system, strengthening the health supervision and safty disposal, training professional personnel and intensifying publicity and education, etc.