Cysteinyl leukotriene receptor antagonist alleviates global cerebral ischemia/reperfusion injury in gerbils through down?regulating autophagy / 中国实验动物学报

Objective Cysteinyl leukotrienes are potent inflammatory mediators. Their actions are mediated by specific receptors,the CysLT receptors(CysLT1R and CysLT2R),which have been cloned and characterized. In this stud-y,we investigated the protective effects of the CysLTR antagonist Pranlukast and HAMI 3379 on global cerebral ischemia/reperfusion(CI/R)injury in gerbils and its underlying mechanisms. Methods The gerbil model of CI/R was established by bilateral common carotid artery occlusion for 10 min followed by 24 h reperfusion. Then the animals were equally ran-domized into four groups: sham, model, Pranlukast(0.1 mg/kg)and HAMI 3379(0.1 mg/kg)groups. The later two groups were treated with intraperitoneal injection of Pranlukast and HAMI 3379,respectively,once daily for 4 days before carotid artery occlusion,while the former two groups with saline only,all at 10 mL/kg. After 24 h reperfusion,neurologi-cal deficit scores were observed and the behavioral dysfunction was assessed. The neuron morphology of cerebral cortex and CA1 subregion of hippocampus were observed in brain sections stained with cresyl violet. The expression of autophagy-relat-ed proteins beclin-1 and LC3 in the homogenate of cerebral cortex and hippocampus were determined using western blotting analysis. The ultrastructure of autophagosomes in the CA1 subregion of hippocampus was observed by electron microscopy. Results Compared with the model group, Pranlukast and HAMI 3379 attenuated neurological deficits, improved the be-havioral dysfunction,inhibited the neuron injury and loss, decreased the expression of autophagy-related protein beclin-1 and LC3 and the number of autophagosomes. Conclusions cysteinyl Leukotriene receptor antagonist Pranlukast and HAMI 3379 can alleviate global cerebral ischemia/reperfusion injury in gerbils. The protective effects of Pranlukast and HAMI 3379 appear to be associated with the inhibition of autophagy.

Preliminarily genetic characterizations of nonalcoholic fatty liver disease mongolian gerbil / 中国比较医学杂志

Non-alcoholic fatty liver disease (NAFLD) has become one of the major metabolic diseases.In view of the defects of traditional animal models, this study was the first to establish the NAFLD model of Mongolian gerbil (Meriones unguiculatus) with simple feed formula which is similar to human (from simple fatty liver to steatohepatitis, fibrosis,Liver cirrhosis).This study discussed the mechanism of rapid fatty liver deposition in Mongolian gerbil, revealed its molecular mechanism,main regulatory target and network function of fatty liver susceptibility.We provide a new animal model of NAFLD with relatively clear background and less time-consuming for clinical treatment and new drug development.The theoretical and practical basis for the breeding of inbred strain NAFLD gerbil was established.

Establishment and evaluation of a high throughput sequencing technology for detection of Salmonella in laboratory animals / 中国比较医学杂志

Objective To establish a detection method of Salmonella in laboratory animals based on a high-throughput sequencing technology, and to apply it in detection of Salmonella in laboratory animals.Methods DNA samples were extracted from mouse feces.Universal primers for 16S rDNA, 23S rDNA, 16S-23S rDNA, 23S-5S rDNA region, gyrB preferred area were designed, respectively.Each primer was tested and analyzed to determine the best amplification conditions and build a database.Forty-two samples of Salmonella were assayed by Illumina high-throughput sequencing technology and evaluated the specificity and stability of this method.Results The species preferred region of Salmonella was gyrB gene region.The primers for gyrB gene were FP5 ’-AACCACCGCAATCAGACCTT3‘ and FP5 ’-AGCCACGAAACCTTCACYA-3’.The primers were optimized and determined, through a high-throughput sequencing, and the sequence analysis detected very small amount of Salmonella in the 42 samples, indicating that this detection method is stable, highly sensitive, and the limit of detection reached to 0-102 CFU.Conclusions We have established a complete detection system for detection of Salmonella in laboratory animals based on a high-throughput sequencing technology, This system can detect trace amounts of Salmonella in laboratory animals, and this detection method is stable and highly sensitive, which can be also used in detection of other kinds of pathogenic microorganism in laboratory animals.

Effect of mitochondrial cytochrome c on hepatocyte apoptosis in non-alcoholic fatty liver disease in rabbits / 中国比较医学杂志

Objective To investigate the role of mitochondrial cytochrome c on hepatocyte apoptosis in non-alcoholic fatty liver disease in rabbits and its pathogenesis.Methods Forty Japanese white rabbits were randomly assigned to control group and model group.The model group was divided into three subgroups: 4-week, 6-week, and 8-week groups, with 10 rabbits in each group.The model groups were subcutaneously injected with peanut oil (1.2 mL/kg), twice a week for 4 weeks, 6 weeks or 8 weeks.The rabbits of all groups were killed at the right time.Serum samples were collected to detect the serum biochemical index levels.Liver tissue samples were taken for pathological observation using HE staining.The hepatocyte apoptosis index ( AI ) was measured by flow cytometry, and mitochondrial permeability transition pore ( MPTP) was evaluated by ultraviolet spectrophotometry.Immunohistochemistry was employed to detect the hepatic expressions of Bcl-2, Bax, CYT C and caspase-3.Western blot was performed to detect the changes of CYT C and caspase-3 protein expressions.Results The model groups showed hepatic injury and high level of TC, TG, CRP, IL-6 and TNF-αbeginning from 4 weeks.With the NAFLD process, the hepatocyte apoptosis index was significantly increased at 4-8 weeks and the MPTP was gradually increased.In the model group, hepatic Bcl-2, Bax, CYT C and caspase-3 expressions were increased steadily with the time passing.Conclusions NAFLD-induced liver damage is associated with apoptosis, and the mitochondrial cytochrome c-mediated apoptotic pathway plays a role in the occurrence of NAFLD.

Efficient preparation of a TXNIP knockout mouse model by transcription activator-like effector nucleases (TALEN) / 中国比较医学杂志

Objective To knockout the murine Txnip gene using microinjection of transcription activator-like effector nuclease ( TALEN) mRNAs.Methods TALEN knockout site recognizing Txnip was designed by tools on line, then constructed the vectors and assayed its cleavage activity at cellular level.TALEN mRNA was transcribed in vitro and microinjected into C57BL/6J mouse zygotes.F0 mice were verified at DNA level with BamHI and TXNIP-knockout mice were obtained.Results We designed and constructed TALENs which recognized and cut the first exon of Txnip, and got four TXNIP knockout mice, among which two were frameshift mutation, demonstrating that the TXNIP-knockout mice were generated by TALEN technique.Conclusions Microinjection of in vitro transcribed TALEN mRNAs into murine zygotes is a highly effective and convenient way to develop TXNIP-knockout mouse model.

PCR sequencing for detection and gene typing of hantavirus isolated from rodents / 中国实验动物学报

Objective To evaluate the application value of PCR-sequencing in clinical detection of hantavirus in rodents .Methods Based on 7 subtypes and 24 strains of representative hantavirus strains downloaded from Genbank , the virus S gene fragments were used for primer design and neighbor joining method was applied for phylogenetic analysis . Thereafter, we identified hantavirus strains isolated from wild rodents in recent years in Zhejiang Province by this method . Results The 24 analyzed strains were divided into 5 regions in the phylogenetic tree .Four of them with topology structure were more stable .Eleven strains of the virus were amplified by PCR and sequenced , and the results showed that the prim-ers were with high sensitivity and specificity .Three HTN strains and 1 strain of serotype SEO were distinguished from 9 strains of unknown strains isolated in Zhejiang Province .We also found that 5 strains of hantavirus belonging to two un-known serotypes .Discussion Our results suggest that the PCR-sequencing method proposed in this study can be used for clinical detection of hantavirus .

Effect of propolis on the expression of CD54 and activation of NF-?B p65 of lung tissue in acute lung injury rats / 中国病理生理杂志

AIM: To study the effect of propolis on the expression of CD54 and activation of NF-?B p65 in lung tissue of acute lung injury (ALI) rats. METHODS: 40 male Wistar rats were divided into 5 groups: normal control, model control, dectancyl group, water soluble derivative of propolis (WSP) group and ethanol extracted propolis (EEP) group. ALI animal model was performed by oleic acid and LPS twice attack. The pathologic slice was observed with light microscope and the NF-?B p65 activity and CD54 expression were tested by immunohistochemistry (SABC and SP). RESULTS: Both EEP and WSP antagonized the lung edema, decreased the inflammation and inhibited the expression of CD54 and activation of NF-?B p65. CONCLUSION: The increase in the expression of CD54 and the activation of NF-?B p65 in the lung tissues of ALI were involved in the formation of ALI. Propolis ameliorated the lung damage, which maybe related to the inhibition of CD54 expression and NF-?B p65 activation.

STUDY ON THE EFFECT OF PROPOLIS EXTRACTS ON ACUTE PLEURISY AND ITS MECHANISM IN RATS / 营养学报

Objective: To investigate the anti-inflammatory effect and the possible mechanism of water and ethanol extracts of propolis. Method: Forty male Wistar rats were divided into 5 groups: normal group, model group, medicine groups, two groups treated with water and ethanol extracts of propolis. The acute pleurisy model was established by injecting carrageenan. The effects of propolis on acute pleurisy was studied by counting leukocytes, measuring the content of MDA, lysozyme and activity of SOD in serum and the content of NO, protein and PGE2 in pleural effusion. Results: The propolis solutions extracted by water and ethanol presented obvious effect on inflammation. It could antagonize the purulent pleurisy, reduce the number of leukocytes and the content of MDA, lysozyme and activity of SOD in serum and the contents of NO, protein and PGE2 and decrease the inflammation. Conclusion: Propolis displays anti-inflammatory effects by decreasing the action of NO and PGE2 and preventing the activation of protein kinase.