MICE

摘要： Objective:To study the regulatory effect of cryoablation on MAPK/ERK signaling pathway in mice with lung adenocarcinoma.Methods:Lewis mouse lung adenocarcinoma cell line was used to establish subcutaneous transplanted tumor model in C57BL/6 mice.Ten mice were randomly divided into two groups:sham operation group and cryoablation group,with 5 mice in each group.The cryoablation group was treated with double circulation-rewarming ablation,and the sham operation group was treated with incision and suture at the transplanted tumor.The tumor tissues were taken 14 days after operation.Detect the effect of cryoablation on MAPK/ERK pathway related proteins by Western blot,such as KRAS,RAF1,MEK1,ERK1/2,P-RAF1,P-MEK1,P-ERK1/2.The expression of KRAS gene was further verified by qRt-PCR.Results:Compared with the sham operation group,the phosphorylated proteins P-RAF1,P-MEK1 and P-ERK1/2 in tumor tissue after cryoablation were decreased(P<0.05),and the key molecule KRAS in MAPK/ERK pathway was decreased in protein and gene expression(P<0.05).Conclusion:Cryoablation can negatively regulate MAPK/ERK signaling pathway by down-regulating KRas expression.

摘要： BACKGROUND: To investigate effects of Maxingloushi decoction on lung inflammation and programmed death markers(programmed death-1 [PD-1], programmed death-ligand 1 [PD-L1]) in the lung tissue, peripheral blood, and bronchoalveolar lavage fl uid(BLF) in a mouse model of chronic obstructive pulmonary disease(COPD).METHODS: Thirty-six mature male BALB/C mice were randomly divided into normal group(group A, n=6), COPD model group(group B, n=10), Maxingloushi decoction + COPD group(group C, n=10), and PD-1 inhibitor + COPD group(group D, n=10). The COPD model was established by smoke inhalation combined with lipopolysaccharide(LPS). Levels of PD-1 and PD-L1 in plasma and BLF were measured by enzyme-linked immunosorbent assay(ELISA). Histopathological techniques were used to semi-quantitatively analyze the immuno-fluorescence optical density(IOD) value of the lung tissue. RESULTS: In plasma and BLF, the expression of PD-1 in the group B was higher than that in the group A, and the expression of PD-L1 was lower than that in the group A. The expression of PD-1 and PD-L1 in the lung tissue was normalized in the group C in comparison with the group B(P<0.05) and the group D(P<0.05), and infl ammatory cell infiltration in the lung tissue was also improved.CONCLUSIONS: These findings reveal that COPD causes an immune imbalance in the peripheral blood and lung tissue, and that both Maxingloushi decoction and PD-1 inhibitor treatment can mitigate lung inflammation in COPD by reducing PD-1 expression and increasing PD-L1 expression. The treatment effect of Maxingloushi decoction may be superior to that of PD-1 inhibitor.

摘要： Interleukin 17A(IL-17A)was previously shown to be a key pro-inflammatory factor in diabetes mellitus and associated complications.However,the role of IL-17A in diabetic encephalopathy remains poorly understood.In this study,we established a mouse model of diabetic encephalopathy that was deficient in IL-17A by crossing Il17a-/-mice with spontaneously diabetic Ins2^(Akita)(Akita)mice.Blood glucose levels and body weights were monitored from 2-32 weeks of age.When mice were 32 weeks of age,behavioral tests were performed,including a novel object recognition test for assessing short-term memory and learning and a Morris water maze test for evaluating hippocampus-dependent spatial learning and memory.IL-17A levels in the serum,cerebrospinal fluid,and hippocampus were detected with enzyme-linked immunosorbent assays and real-time quantitative polymerase chain reaction.Moreover,proteins related to cognitive dysfunction(amyloid precursor protein,β-amyloid cleavage enzyme 1,p-tau,and tau),apoptosis(caspase-3 and-9),inflammation(inducible nitric oxide synthase and cyclooxygenase 2),and occludin were detected by western blot assays.Pro-inflammatory cytokines including tumor necrosis factor-α,interleukin-1β,and interferon-γin serum and hippocampal tissues were measured by enzyme-linked immunosorbent assays.Microglial activation and hippocampal neuronal apoptosis were detected by immunofluorescent staining.Compared with that in wild-type mice,mice with diabetic encephalopathy had higher IL-17A levels in the serum,cerebrospinal fluid,and hippocampus;downregulation of occludin expression;lower cognitive ability;greater loss of hippocampal neurons;increased microglial activation;and higher expression of inflammatory factors in the serum and hippocampus.IL-17A knockout attenuated the abovementioned changes in mice with diabetic encephalopathy.These findings suggest that IL-17A participates in the pathological process of diabetic encephalopathy.Furthermore,IL-17A deficiency reduces diabetic encephalopathy-mediated neuroinflammation and cognitive defects.These results highlight a role for IL-17A as a mediator of diabetic encephalopathy and potential target for the treatment of cognitive impairment induced by diabetic encephalopathy.

摘要： There is a critical need to develop animal models to alleviate vaccine and drug development difficulties against zoonotic viral infections.The coronavirus family,which includes severe acute respiratory syndrome coronavirus 1 and severe acute respiratory syndrome coronavirus 2,crossed the species barrier and infected humans,causing a global outbreak in the 21st century.Because humans do not have pre-existing immunity against these viral infections and with ethics governing clinical trials,animal models are therefore being used in clinical studies to facilitate drug discovery and testing efficacy of vaccines.The ideal animal models should reflect the viral replication,clinical signs,and pathological responses observed in humans.Different animal species should be tested to establish an appropriate animal model to study the disease pathology,transmission and evaluation of novel vaccine and drug candidates to treat coronavirus disease 2019.In this context,the present review summarizes the recent progress in developing animal models for these two pathogenic viruses and highlights the utility of these models in studying SARS-associated coronavirus diseases.

摘要： The effect of Fuzhuan brick tea(FBT)on metabolism in obese mice is mediated by regulation of N-methyltransferase by aryl hydrocarbon receptor.The expression of the phosphatidylethanolamine N-methyltransferase gene is regulated by many transcription factors,and those specific to this effect need further investigation.Experimental animal studies have been designed to observe the effects of a single drug or the sequential effects of drugs.A washout period should be included if different drugs(e.g.,antibiotics and FBT)are given to avoid or reduce additive effects or synergy.Currently,most experimental studies performed in mice used only male animals.However,experience has revealed that the results of using only male mice are very likely to have sex differences.

摘要： Objective:To characterize the antifungal activity of methanolic leaf extract of Calotropis gigantea alone or in combination with amphotericin B against invasive pulmonary aspergillosis in mice.Methods:GC/MS was used for analysis of active constituents of Calotropis gigantea extract.Spore germination assay and broth micro-dilution method were used to determine antifungal potential of Calotropis gigantea/amphotericin B against Aspergillus fumigatus.Neutropenic mice were randomly assigned into 5 groups:group 1 was neutropenic(control);group 2 was infected with Aspergillus fumigatus;group 3 was infected with Aspergillus fumigatus,and treated with Calotropis gigantea extract;group 4 was infected with Aspergillus fumigatus and treated with amphotericin B;group 5 was infected with Aspergillus fumigatus and treated with both Calotropis gigantea extract and amphotericin B.Fresh lung tissues were histopathologically examined.Fungal burden and gliotoxin concentration were evaluated in lung tissues.Catalase,superoxide dismutase,and malondialdehyde content were determined in lung tissues.Myeloperoxidase,tumor necrosis factor-alpha,interleukin-1,and interleukin-17 were also estimated by the sandwich enzyme-linked immuno-sorbent assay.Results:Calotropis gigantea/amphotericin B had a minimum inhibitory concentration and minimum fungicidal concentration of 80 and 160μg/mL,respectively,for Aspergillus fumigatus.Additionally,Calotropis gigantea/amphotericin B significantly reduced lung fungal burden by 72.95%and inhibited production of gliotoxin in lung tissues from 6320 to 1350μg/g lung.Calotropis gigantea/amphotericin B reduced the oxidative stress of the lung via elevating the activity of antioxidant enzymes and decreasing the levels of lipid peroxidation.Myeloperoxidase activity and the production of pro-inflammatory cytokines were also significantly reduced.Scanning electron microscopy revealed deteriorations in the hyphae ultrastructure in Calotropis gigantea/amphotericin B treated Aspergillus fumigatus and leak of cellular components after damage of the cell wall.In vivo study revealed the suppression of lung tissue damage in mice of invasive pulmonary aspergillosis,which was improved with Calotropis gigantea/amphotericin B compared to the control group.Conclusions:Calotropis gigantea/amphotericin B is a promising treatment to reduce lung fungal burden and to improve the drugs’therapeutic effect against invasive pulmonary aspergillosis.

摘要： Recent studies have demonstrated a new role for Klf10, a Krüppel-like transcription factor, in skeletal muscle, specifically relating to mitochondrial function. Thus, it was of interest to analyze additional tissues that are highly reliant on optimal mitochondrial function such as the cerebellum and to decipher the role of Klf10 in the functional and structural properties of this brain region. In vivo (magnetic resonance imaging and localized spectroscopy, behavior analysis) and in vitro (histology, spectroscopy analysis, enzymatic activity) techniques were applied to comprehensively assess the cerebellum of wild type (WT) and Klf10 knockout (KO) mice. Histology analysis and assessment of locomotion revealed no significant difference in Klf10 KO mice. Diffusion and texture results obtained using MRI revealed structural changes in KO mice characterized as defects in the organization of axons. These modifications may be explained by differences in the levels of specific metabolites (myo-inositol, lactate) within the KO cerebellum. Loss of Klf10 expression also led to changes in mitochondrial activity as reflected by a significant increase in the activity of citrate synthase, complexes I and IV. In summary, this study has provided evidence that Klf10 plays an important role in energy production and mitochondrial function in the cerebellum.

摘要： Nuclear factor erythroid-derived 2-like 2(Nrf2)is the master regulator of antioxidant defenses.High-intensity interval training(HIIT)has been proposed as a time-efficient training program and has become a substantial component of modern training program In the present study,we evaluated the effects of sulforaphane(SFN),a dietary isothiocyanate derived from cruciferous vegetables and a potent Nrf2 activator,on Nrf2-mediated antioxidant defense responses of skeletal muscle induced by exhaustive exercise in HIIT mice.Male C57 BL/6 J mice were randomly allocated into control group,HIIT group,and HIIT pretreated with SFN(HIIT+SFN)group.On the third day after completion of a 6-weeks HIIT protocol,an exhaustive treadmill test was conducted in all mice.Mice were intraperitoneally injected with SFN(HIIT+SFN group)or PBS(HIIT and control mice)4 times in 3 days prior to the exhaustive treadmill test.The results indicated that the 6-weeks HIIT protocol did not increase the antioxidative capacity of skeletal muscle during exhaustive exercise.Importantly,SFN treatment improved anti oxidative capacity of skeletal muscle in response to the acute exhaustive exercise by increasing mRNA and nucleoprotein expression of Nrf2 and these genes involved in antioxidant generation and decreasing blood creatine kinase(CK)and 4-hydroxy-2-nonenal(4-HNE)-modified protein levels in the HIIT mice.

摘要： Objective:To evaluate the antioxidant potential and pancreatic lipase inhibitory action of optimized hydroethanolic extracts of Solanum nigrum.Methods:Optimized extraction for maximum recovery of metabolites was performed using a combination of freeze-drying and ultrasonication followed by determination of antioxidant and antiobesity properties.The ultra-high performance liquid chromatography equipped with mass spectrometry was used to analyze metabolite profiling of Solanum nigrum.Computational studies were performed using molecular docking and electrostatic potential analysis for individual compounds.The hypolipidemic potential of the most potent extract was assessed in the obese mice fed on fat rich diet.Results:The 80%hydroethanolic extract exhibited the highest extract yield,total phenolic contents,total flavonoid contents along with the strongest 2,2-diphenyl-1-picrylhydrazyl scavenging activity,total antioxidant power,and pancreatic lipase inhibitory properties.The 80%hydroethanolic extract not only regulated the lipid profile of obese mice but also restricted the weight gain in the liver,kidney,and heart.The 80%hydroethanolic extract also reduced alanine transaminase and aspartate transaminase concentrations in serum.The effects of plant extract at 300 mg/kg body weight were quite comparable with the standard drug orlistat.Conclusions:Solanum nigrum is proved as an excellent and potent source of secondary metabolites that might be responsible for obesity mitigation.

摘要： Objective:To investigate the effect of Oroxylum indicum fruit extract on high-fat diet-induced hyperlipidemic mice.Methods:The phytochemical composition of Oroxylum indicum fruit extract was determined by liquid chromatographymass spectrometry/mass spectrometry(LC-MS/MS)and gas chromatography-mass spectrometry.Forty-two male mice were used.The mice were divided into six groups:normal control,high-fat diet control,simvastatin treatment(20 mg/kg BW/day),and Oroxylum indicum fruit extract(100,200,300 mg/kg BW/day)treatment groups.Food intake,body weight,serum parameters,lipid profile,and histopathological lesions of the kidney,liver,and epididymal fat were observed.Results:LC-MS/MS results revealed four major components of Oroxylum indicum fruit extract:luteolin,apigenin,baicalein,and oroxylin A.Twenty-seven volatile oils were identified from Oroxylum indicum fruit extract.Daily oral administration of Oroxylum indicum fruit extract at 100 to 300 mg/kg BW/day significantly reduced the body weight,total cholesterol,triglyceride,and low-density lipoprotein cholesterol level(P<0.05),whereas high-density lipoprotein cholesterol was higher than the high-fat diet control group.Treatment with 300 mg/kg BW/day Oroxylum indicum fruit extract reduced the pathological lesion and prevented fat accumulation in the kidney and liver.Conclusions:Oroxylum indicum fruit extract has hypolipidemic effect in hyperlipidemic mice,and the active ingredients of Oroxylum indicum fruit extract,both flavonoids and volatile oils,should be further explored as an antihyperlipidemic agent.

摘要： The present study was undertaken to compare the viability and infectivity ofCryptosporidiumparvum oocysts that had been stored for 1,4,7,10,13,16,20,25 and 30 months at4°C in 2.5% potassium dichromate (Cr) and chlorinated tap water,respectively. An excystationprotocol was performed in vitro to evaluate viability. One hundred and eighty female BABL/c micewere used to evaluate the infectivity of oocysts by detecting the prepatent period of C. parvuminfection,the quantity of oocysts excreted,and the number of parasites that colonized the villi of theileum. The results showed that C. parvum oocysts preserved in Cr for 1-16 months and in water for1-13 months were capable of excystation in vitro and infection of mice in vivo. The excystation ratesof oocysts and the prepatent periods in mice infected by oocysts stored in Cr and water were notsignificantly different (p>0.05),and there was a strong correlation between prepatent period andduration ofoocyst storage (Cr: Re=0.92; water. R2=0.98). There were no significant differences inoocyst shedding from feces or parasitism of the terminal ilea of mice by Cryptosporidia between thetwo storage media (p>0.05). In conclusion,C. parvum oocysts may be stored at 4°C in water insteadof Cr for the purposes of laboratory research. However,the presence of viable C. parvum oocysts inwater is a severe challenge to the drinking water treatment industry.

摘要： The experiment was conducted to compare the effect of different selenium sourceson the expression of glutathione peroxidase 1 (GPx 1) and iodothyronine deiodinase 1 (Dio 1) mRNAin mice by quantitative real time PCR. A total of sixty male Kunming mice at average body weightof 20g were allotted to three groups in a randomized complete block design,including twotreatments and one control. Mice in Group 1 were fed a basal diet as control,while mice in Groups2 and 3 were fed the basal diet supplemented with 0.1 mg/kg selenium as sodium selenite orselenized yeast,respectively. Whole feeding experiment lasted for 30 d. At the end of the feedingtrial,liver mRNA levels of GPxl and Diol were determined by quantitative real time PCR,as wellas growth performance,body composition,blood and GPx activity were determined. The resultsshowed that no significant differences in overall growth performance and body composition,including body weight,body length,heart weight,kidney weight and liver weight,were foundbetween the experimental groups (P > 0.05). Blood GPx activity increased in all of the seleniumsupplemented groups compared with control group (P 0.05). In conclusion,selenium increased the mRNA expression of GPxland Diol genes in murine liver,and there was no significant difference between the organic orinorganic form of selenium used.

摘要： 本发明属于生物工程技术领域，具体涉及一种抗Mical2多克隆抗体及其制备方法。该抗Mical2多克隆抗体的制备方法包括如下步骤：构建含有Mical2基因的重组表达载体，所述Mical2基因的核苷酸序列如SEQ ID No.1所示；将所述重组表达载体转化至大肠杆菌感受态细胞中诱导表达，获得Mical2蛋白抗原；将所述Mical2蛋白抗原免疫动物，获得抗血清，然后分离纯化获得抗Mical2多克隆抗体。该制备方法得到的抗Mical2多克隆抗体可以检测Mical2的蛋白水平，还可以用来检测Mical2的组织类型或细胞类型特异性，在细胞或组织中准确定位Mical2的表达位置。

摘要： 本发明属于生物工程技术领域，具体涉及一种抗Mical2多克隆抗体及其制备方法。该抗Mical2多克隆抗体的制备方法包括如下步骤：构建含有Mical2基因的重组表达载体，所述Mical2基因的核苷酸序列如SEQ ID No.1所示；将所述重组表达载体转化至大肠杆菌感受态细胞中诱导表达，获得Mical2蛋白抗原；将所述Mical2蛋白抗原免疫动物，获得抗血清，然后分离纯化获得抗Mical2多克隆抗体。该制备方法得到的抗Mical2多克隆抗体可以检测Mical2的蛋白水平，还可以用来检测Mical2的组织类型或细胞类型特异性，在细胞或组织中准确定位Mical2的表达位置。

摘要： 本发明属于生物工程技术领域，具体涉及一种脑组织中与Mical2相互作用蛋白的鉴定方法。包括如下步骤：构建鼠脑组织mRNA文库；构建含有Mical2基因的融合表达载体，所述Mical2基因的核苷酸序列如SEQ ID No.1所示；利用酵母双杂交技术，将所述融合表达载体转化到感受态酵母中进行培养，然后与含有所述鼠脑组织mRNA文库的文库酵母进行混合培养；将所述混合培养后的培养液涂板，挑选单克隆菌落，筛选出与Mical2蛋白相互作用的蛋白质。通过该鉴定方法，在鼠脑中发现了125种可以与Mical2相互作用的蛋白质，这为进一步研究Mical2在脑组织中的功能指出了方向。

摘要： 本发明涉及一种蛋氨酸亚砜氧化酶MICAL1突变蛋白及其应用，通过将大量癌症患者作为研究病例，对病例进行基因检测并分析，确定出人的蛋氨酸亚砜氧化酶MICAL1的突变蛋白，根据该人的蛋氨酸亚砜氧化酶MICAL1突变蛋白制备基因芯片、单克隆抗体和ELISA试剂盒，为癌症的基因诊断提供指引作用，实现癌症的早诊断、早发现和相关治疗。

摘要： 本实用新型涉及一种三合一的Micor USB接口应用于手机的电路，该电路的Micor USB 5PIN数据线端的PIN1(VUSB)口与uart口下载线、USB口下载线的PIN1连接，Micor USB 5PIN数据线端的PIN2(D-)口与耳机的SPK_L/FM天线、USB口下载线的USB_DM口连接，Micor USB 5PIN数据线端的PIN3口(D+)与耳机的SPL_R口、uart口下载线的TX口、USB口下载线的USB_DP口连接，本实用新型将USB、耳机、下载做到三合一，下载可支持uart口和USB下载，能够节省一个耳机座。

摘要： 本发明公开了一种基于流表虚拟化的mice flow聚合方法，在OpenFlow环境下基于特征项和偏好的流聚合策略，通过该方式能够结合网络设备偏好和mice flow的状态动态调节流聚合策略，达到了更高的流聚合效率。同时，本发明利用虚拟数据流标识符和虚拟流表实现mice flow的聚合，决策行为由OpenFlow控制器完成，只需要依据虚拟数据流标识符就可以指示网络设备完成流聚合行为，将传统的流聚合控制面剥离到控制器。本发明在减少数据流个数同时符合网络设备个性化要求，从而提高了OpenFlow网络的性能。

摘要： 本发明公开了一种基于流表虚拟化的mice flow聚合方法，在OpenFlow环境下基于特征项和偏好的流聚合策略，通过该方式能够结合网络设备偏好和mice flow的状态动态调节流聚合策略，达到了更高的流聚合效率。同时，本发明利用虚拟数据流标识符和虚拟流表实现mice flow的聚合，决策行为由OpenFlow控制器完成，只需要依据虚拟数据流标识符就可以指示网络设备完成流聚合行为，将传统的流聚合控制面剥离到控制器。本发明在减少数据流个数同时符合网络设备个性化要求，从而提高了OpenFlow网络的性能。

摘要： 本发明涉及生物医药领域，特别是涉及Rolling Nagoya和Leaner mice突变小鼠模型大脑内Ca