Establishment and behavioral evaluation of a mouse model of long-term free-choice alcohol drinking / 北京大学学报(医学版)

OBJECTIVE@#To establish a model of long-term free drinking mouse by feeding mice with alcohol to simulate the state of human voluntary long-term drinking, and on this basis, to further discuss the evaluation criteria of long-term free drinking mice model in sports, anxiety and cognitive behavior.@*METHODS@#Forty six-week-old SPF C57BL/6 male mouse were randomly divided into two groups: Long-term free drinking group (n=20) and normal control group (n=20). The two groups were given solid feed normally. The long-term free drinking group was free to take 10% alcohol and water every day, while the normal drinking group only took water every day. The mice were fed for 7 months, and were evaluated by a series of behavioral methods, including Rota-rod test, balance beam test, open filed test, the elevated plus maze, two-box social behavior, new object recognition, Y maze and water maze.@*RESULTS@#With the increase of drinking days, the mice showed significant alcohol addiction in the alcohol preference test. With the increase of alcohol intake, the mice in the long-term free choice drinking group had slightly shiny fur and reduced diet. Compared with the control group, the weight gain began to slow down from the third month, and the weight decreased significantly by the sixth and seventh months (P=0.006, P < 0.001). The mice showed reduced balance locomotion ability (P=0.003, P=0.001) in the rotary bar and balance beam test. In the open field and elevated cross test, the mice had obvious anxiety-like behavior (P < 0.001). The mice showed decreased social ability in the two boxes of social behavior (P < 0.016). In the experiment of new object recognition and Y maze, the exploration of new object decreased (P=0.018, P=0.040). In the water maze, cognitive functions, such as learning and spatial memory were reduced (P < 0.001).@*CONCLUSION@#The successful establishment of the long-term free drinking mouse model is more convenient for us to carry out further research on the neural mechanism of alcohol addiction, and lays an experimental foundation for exploring the neural mechanism of alcohol addiction and related new targets.

Study on the modeling method and pathological parameters of chronic atrophic gastritis / 中华消化杂志

Objective:To explore an ideal method for establishing a mouse model of chronic atrophic gastritis (CAG).Methods:CAG mouse models were established with five different modeling methods ( N-methyl- N′-nitro- N-nitrosoguanide (MNNG), sodium salicylate, sodium deoxycholate, Helicobacter pylori infection, and combinations of them) in BALB/c and C57 mice. The effect of each modeling method was evaluated by histological observation of gastric mucosa, plasma biochemical parameters, inflammatory response score, and the expression of anti-inflammatory factors. Results:The results of histological observation of gastric mucosa showed that all of the 5 methods could successfully establish CAG mouse models. In BALB/c mice, compared with the healthy control group, significant features of CAG accompanied with intestinal metaplasia was found in the model group established by combination of MNNG-free drinking, 2% sodium salicylate and 20 mmol sodium deoxycholate. From the results of serological detection, compared with the normal control group, the mRNA expression levels of related anti-inflammatory factors interleukin-2, interleukin-10, interleukin-13 and growth differentiation factor-15 of each model group decreased, which indicated that the mice of each CAG model group had different degrees of inflammation. The results of plasma biochemical parameters indicated that plasma gastrin of each group decreased and the ratio of pepsinogen Ⅰ and pepsinogen Ⅱ significantly dropped. The above results demonstrated that in BLAB/c mice, MNNG-free drinking, 2% sodium salicylate and 20 mmol sodium deoxycholate was better than other four modeling methods. For C57 mice, it was also found that simple chemical drug mutagenesis and Helicobacter pylori replication method both could successfully establish CAG models. No matter from pathological observation, relative expression of anti-inflammatory factors and analysis of plasma biochemical parameters, the effects of combination of the two methods was better. Conclusion:The CAG mouse model established by MNNG-free drinking, 2% sodium salicylate and 20 mmol sodium deoxycholate can provide a certain reference for the establishment and application of mouse model in CAG experiments in the future for pharmacological research.

Pathogenic role of NDUFA13 inactivation in spontaneous hepatitis in mice and the mechanism / 南方医科大学学报

OBJECTIVE@#To investigate the role of NDUFA13 inactivation in the pathogenesis of spontaneous hepatitis in mice and explore the possible mechanisms.@*METHODS@#Hepatocyte-specific NDUFA13 knockout (NDUFA13@*RESULTS@#Liver-specific NDUFA13 heterozygous knockout mice were successfully constructed as verified by PCR results. HE staining revealed severe liver damage in both 4- week-old and 2-year-old NDUFA13@*CONCLUSIONS@#Hepatocytes-specific NDUFA13 ablation can trigger spontaneous hepatitis in mice possibly mediated by the activation of ROS/NF-κB/NLRP3 signaling.

Metabolite profiling and antidiabetic attributes of ultrasonicated leaf extracts of Conocarpus lancifolius

Objective: To profile the secondary metabolites and to evaluate the antidiabetic potential of hydroethanolic leaf extracts of Conocarpus lancifolius.Methods: The various hydroethanolic extracts o f Conocarpus lancifolius leaf were prepared by ultrasonication assisted freeze-drying. Total phenolic contents, flavonoid contents, antioxidant activity, α-glucosidase and α-amylase inhibitions of leaf extracts were determined. The metabolite profiling was accomplished by UHPLC-Q-TOF-MS/MS analysis. The antidiabetic assessment of the most potent extract was carried out by measuring the hypoglycemic and hypolipidemic effect in the high fat diet-fed diabetic albino mice. The blood glucose level, haemoglobin, total cholesterol, high-density lipoproteins (HDL) and low-density lipoproteins (LDL) were determined. Results: The 60％ ethanolic extract exhibited the highest phenolic and flavonoid contents of (349.39 ± 2.13) mg GAE/g dry extract and (116.95 ± 2.34) mg RE/g dry extracts, respectively, and the highest DPPH scavenging activity with an IC50 value of (32.87 ± 1.11) μg/mL. The IC50 values for α-glucosidase and α-amylase inhibitions were (38.64 ± 0.93) μg/mL and (44.80 ± 1.57) μg/mL, respectively. UHPLC-Q-TOF-MS/MS analysis confirmed the presence of gallic acid, ellagic acid, corilagin, kaempherol-3-O-rutinoside, caffeic acid derivative, isorhamnetin and galloyl derivatives in the 60％ ethanolic extract. Plant extract at a dose of 450 mg/kg body weight reduced blood glucose level, total cholesterol, LDL and HDL, and increased haemoglobin in alloxan-induced diabetic mice, Conclusions: Conocarpus lancifolius leaves are proved as a good source of biologically functional metabolites and possess antidiabetic activity which may be further explored to treat diabetes.

Metabolite profiling and antidiabetic attributes of ultrasonicated leaf extracts of Conocarpus lancifolius

Objective: To profile the secondary metabolites and to evaluate the antidiabetic potential of hydroethanolic leaf extracts of Conocarpus lancifolius. Methods: The various hydroethanolic extracts of Conocarpus lancifolius leaf were prepared by ultrasonication assisted freeze-drying. Total phenolic contents, flavonoid contents, antioxidant activity, α-glucosidase and α-amylase inhibitions of leaf extracts were determined. The metabolite profiling was accomplished by UHPLC-Q-TOF-MS/MS analysis. The antidiabetic assessment of the most potent extract was carried out by measuring the hypoglycemic and hypolipidemic effect in the high fat diet-fed diabetic albino mice. The blood glucose level, haemoglobin, total cholesterol, high-density lipoproteins (HDL) and low-density lipoproteins (LDL) were determined. Results: The 60% ethanolic extract exhibited the highest phenolic and flavonoid contents of (349.39 ± 2.13) mg GAE/g dry extract and (116.95 ± 2.34) mg RE/g dry extracts, respectively, and the highest DPPH scavenging activity with an IC50 value of (32.87 ± 1.11) μg/mL. The IC50 values for α-glucosidase and α-amylase inhibitions were (38.64 ± 0.93) μg/mL and (44.80 ± 1.57) μg/mL, respectively. UHPLC-Q-TOF-MS/MS analysis confirmed the presence of gallic acid, ellagic acid, corilagin, kaempherol-3-O-rutinoside, caffeic acid derivative, isorhamnetin and galloyl derivatives in the 60% ethanolic extract. Plant extract at a dose of 450 mg/kg body weight reduced blood glucose level, total cholesterol, LDL and HDL, and increased haemoglobin in alloxan-induced diabetic mice, Conclusions: Conocarpus lancifolius leaves are proved as a good source of biologically functional metabolites and possess antidiabetic activity which may be further explored to treat diabetes.

Mechanism of Duanteng Yimu Decoction on Expression of MEKK2 in Rheumatoid Arthritis Synovial Fibroblasts and Arthritis in CIA Mice / 中国实验方剂学杂志

Objective::Duanteng Yimu decoction(DTYMD)is effective in treatment of rheumatoid arthritis (RA) by relieving joint inflammation and down-regulating some inflammatory factors in a short period of time, but the mechanism is still unclear. We aimed to investigate upstream kinase of mitogen activated protein kinases(MAPK) and define the anti-inflammatory mechanism of DTYMD. Method::Fibroblasts-like synovial cells(FLSs) were divided into blank group, model group (IL-1β), high-dose DTYMD group (1 000 mg·L-1), medium-dose DTYMD group (800 mg·L-1), low-dose DTYMD group (600 mg·L-1) and armour ammonia butterfly(MTX) group (20 μmol·L-1). The protein and mRNA expressions of mitogen-activated protein kinase kinase kinase 2 (MEKK2) were analyzed by real-time fluorescence quantitative PCR(Real-time PCR). Totally 42 male DBA/1J mice were randomly divided into 6 groups, with 7 mice in each group, namely normal group, model group and MTX group (2 mg·kg-1), low-dose DTYMD group (6.25 mg·kg-1), medium-dose DTYMD group (12.5 mg·kg-1), and high-dose DTYMD group (25 mg·kg-1). Except for the normal group, the other five groups were included in collagen-induced arthritis(CIA) model by secondary immunoassay. After administration, the posterior limbs and ankle joints were stained with htoxylin-eosin(HE), and the pathological scores of the joints were evaluated. Result::Compared with the model group, DTYMD inhibited the activity of FLSs in a concentration-dependent manner (P<0.01). Compared with the blank control group, the cell proliferation rate of the model group increased (P<0.01). Compared with the model group, high and middle-dose DTYMD groups could inhibit protein and mRNA expressions of MEKK2 (P<0.01), but there was no significant difference in low-dose group. However, the expression of DTYMD protein in high/medium/low-dose groups was significantly higher than that in blank group (P<0.01), but there was no significant difference in MTX group. Compared with the model group, the expressions of matrix metalloprotease-1 (MMP-1), tumor necrosis factor-α(TNF-α) and interleukin(IL)-6 were negatively regulated in different DTYMD groups(P<0.01), and the expressions of MMP-1, IL-6, TNF-α in the model group were significantly higher than those in the blank group (P<0.05, P<0.01). In the animal experiment, compared with the model group, high/middle-dose DTYMD groups could decrease the degree of joint swelling in CIA mice (P<0.01), but there was no significant difference in the low dose group, and the joint swelling in the model group was significantly higher than that in the blank group (P<0.05). In HE staining of ankle joint of CIA mice, the pathological scores of high/small-dose DTYMD groups were significantly lower those of model group (P<0.05, P<0.01), and the pathological score of model group was higher than that of blank group (P<0.01). Conclusion::DTYMD might down-regulate MEKK2 to negatively regulate inflammatory cytokines IL-6, TNF-α and MMP-1, thereby alleviating the inflammatory response in rheumatoid arthritis.

Therapeutic Effect and Mechanism of Chaige Qinlian Tang on Young Mouse MRSA Pneumonia Model / 中国实验方剂学杂志

Objective::To explore the therapeutic effect and mechanism of Chaige Qinlian Tang on pneumonia in young mice. Method::The pneumonia model was duplicated by slowly dripping Staphylococcus aureus into the nasal cavity of mice.After successful modeling, the mice were randomly divided into model group, clindamycin group, and high and low-dose Chaige Qinlian Tang groups, with sham operation group as negative control group.The rats were given 200 mg·kg-1 high-dose Chaige Qinlian Tang, 100 mg·kg-1 low-dose Chaige Qinlian Tang and 120 mg·kg-1 clindamycin.The mice were observed every day.Colonies were counted in the lungs of each group five days later.The expression levels of interleukin(IL)-16, tumor necrosis factor (TNF)-α in lung lavage fluid of each group were determined by enzyme linked immunosorbent assay (ELISA). Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to measure the expression levels of IL-16, TNF-α in lung lavage fluid of each group.The expressions of tumor necrosis factor receptor (TNFR) 1, Caspase-3 and Caspase-7 in lung and the pathological changes of lung were observed. Result::Compared with the sham operation group, the respiratory state and the activity state of the model mice were worse, and the survival rate was higher in the high-dose Chaige Qinlian Tang group.Compared with the sham operation group, the pulmonary colony counts in the model group and treatment groups were increased, compared with the model group, the lung colony counts in clindamycin group and high-dose Chaige Qinlian Tang group were improved significantly (P<0.05, P<0.01). Compared with the control group, the expression levels of IL-16, TNF-α, TNFR1, Caspase-3, Caspase-7 mRNA and protein in the lung of model group and treatment groups were significantly increased (P<0.01). Compared with model group, the expression levels of IL-16, TNF-α and TNFR1, Caspase-3, Caspase-7 in the lung of clindamycin group and high and low-dose Chaige Qinlian Tang groups were significantly increased (P<0.01). The expression levels of protein and mRNA were significantly decreased (P<0.05, P<0.01), and the pathological changes of lung were improved, especially in clindamycin group and high-dose Chaige Qinlian Tang group. Conclusion::Chaige Qinlian Tang has a certain therapeutic effect on Staphylococcus aureus pneumonia in young mice.This effect may be related to regulating TNFR1, Caspase-3 and Caspase-7 pathways, reducing the secretion of IL-16 and TNF-alpha, and enhancing the clearance of staphylococcus aureus.

Establishment of premature ovarian insufficiency kidney deficiency and blood stasis pattern mouse model with Tripterygium wilfordii polyglycoside for Bushen Culuan Decoction therapy / 中国中药杂志

To establish a mouse model of premature ovarian insufficiency( POI) with kidney deficiency and blood stasis pattern by Tripterygium wilfordii polyglycoside( TWP) gavage,and to evaluate the ovarian function and fertility of the model,in order to find Bushen Culuan Decoction therapeutic mechanism. 60 SPF level Blab/c female mice with normal estrous cycle were randomly divided into 6 groups of 10 each: blank group 1( BG1),blank group 2( BG2),blank fertility group( BFG),model group( MG),model recovery group( MRG) and model fertility group( MFG). The mice in three model groups were treated by gastric gavage with TWP suspension 40 mg·kg-1 twice a day for 14 days,while the mice in three blank groups were treated by gastric gavage with same volume normal saline for 14 days. The mice in BG1 and MG were sacrificed and dissected on day 15. The mice in BG2,BFG,MRG and MFG were returned normal feeding from day 15 and were sacrificed and dissected on day 29. The mice in BFG and MFG were cohabited with male mice with a ratio of 2 ∶1( female ∶male) from day 15. The general situation and estrous cycles of all mice were observed every day. Serum sex hormone levels,ovarian index,uterine index,ovarian morphology,follicle count,ovarian VEGF and ES index were observed within the mice in BG1,BG2,MG and MRG. Pregnancy rate,litter size,survival number of newborn mice and male-female proportion were reported within the mice in BFG and MFG. In model establishing stage,the body weight of mice significantly decreased( P <0. 05) in MG and MFG. Compared with BG1,the mice in model group had irregular estrous cycle,decreased ovarian and uterine indexes,less primordial and developing follicles,more atretic follicles,increased VEGF expression and decreased ES expression( P <0. 05). Compared with blank group 2,the mice in model recovery group had irregular estrous cycle,increased FSH level,decreased ovarian indexes,less primordial and developing follicles,more atretic follicles,increased VEGF expression( P<0. 05). Compared with blank fertility group,the mice in model fertility group had smaller litter size and newborn mice survival count( P<0. 05). Gastric gavage with TWP 40 mg·kg-1 twice a day for 14 days is a feasible way to establish a POI kidney deficiency and blood stasis pattern mouse model. The mice ovarian functions didn't recovery on day 14 after stopping TWP intervening,which could suggest the effectiveness of subsequent therapeutic intervention.

Blockage of macrophage-mediated PD1/PD-L1 pathways inhibits tuberculosis relapse in mice / 中国比较医学杂志

Objective To study the immune intervention effect and mechanism of blockage of macrophage-mediated PD1 /PD-L1 pathways with functional PD-L1(programmed cell death ligand-1,PD-L1)monoclonal antibody upon tuberculosis(TB)relapse in mice. Methods Female C57BL/6 mice were infected by tail vein injection of 106CFU M. tuberculosis H37Rv to obtain active TB infection. Two weeks postinfection, the mice in different groups were administered isoniazid(10 mg/kg)(group ISO)and isoniazid combined with PD-L1 monoclonal antibody(50 μg/each)(group ISO+PD-L1)respectively,continued for four weeks to obtain latent infection. The subsequent relapse was monitored. Among the treatment groups,the TB relapse was induced by TNF-α antibody(50 ug/each)for four weeks from the beginning of latent stage. At each scheduled time point, bacterial loads and pathological changes in the lung, spleen and liver were quantitatively analyzed,thereby,the in vivo intervention effect of PD-L1 monoclonal antibody on tuberculosis recurrence in mice was revealed. The in vitro experiment was further explored whether knock-down the expression of PD-L1 on the infected macrophages could accerlate the macrophage apoptosis. Results The bacterial burden reached 3-4 Lg(CFU/mL),and granuloma lesions were extensive in the lung, spleen and liver in the all infected groups, which appeared as active TB stage at 2nd week postinfection. After treated,the bacterial burden of the lung,spleen and liver was decreased, and the pathological lesions alleviated in the group ISO and group ISO+PD-L1, compared with the model control group, showing significant differences, but there was no significant difference between the two treatment groups. However, compared with the group ISO,the group ISO+PD-L1 had a significantly lower bacterial load and milder pathological lesions during the relapse period. Futhermore, knock-down the expression of PD-L1 on macrophages with anti-PD-L1 or PD-L1-siRNA promoted apoptosis in macrophages. Conclusions Blockade of the PD1/PD-L1 pathway by PD-L1 functional antibody can inhibit TB relapse in mice,and knock-down the expression of PD-L1 on macrophages or PD1/PD-L1 pathway with functional antibody can promote apoptosis in macrophages,which together indicate that PD-L1 blockage can effectively promote isoniazid treatment of TB and remarkably inhibit the recurrence of TB in mice.

The protective effects of WIN55212-2 for lung injury in paraquat poisoned mice / 中华急诊医学杂志

Objective To explore the protective effects of cannabinoid analogues WIN55212-2 on paraquat poisoned mice. Methods Totally 35 healthy male C57BL/6 mice were randomly(random number) divided into four groups: PQ group (paraquat poisoned, n=10), WIN 1 mg group (PQ+WIN55212-21 mg n=10), WIN 2 mg group (PQ+WIN55212-22 mg, n=10), control group (n=5).The PQ poisoned animal models were established in the PQ group, WIN 1 mg group and WIN 2 mg group by intraperitoneally injection of paraquat with a concentration of 20 mg/kg. Intraperitoneal injection of WIN55212-2 (containing Tween 80 cosolvent) at the concentration of 1 mg/kg and 2 mg/kg was performed 1 h before PQ exposure in the two interfered groups. Equivalent volume of saline was given to the control group. WIN55212-2 was injected twice a week from the second week. In the acute phase (14 d), 5 mice were randomly sacrificed in the PQ group, WIN 1 mg group and WIN 2 mg group, and 3 mice were sacrificed in the control group to obtain blood sample, bronchoalveolar lavage fluid (BALF) and lung tissue. All the remaining mice were executed on day 28, and the tissue samples were collected as mentioned above. HE staining and Masson staining were performed to observe the changes of lung tissues after PQ poisoning. Changes of TNF-α, IL-6 and TGF-β in plasma and BALF were measured by ELISA. Results In the acute phase, the pathological sections of lung tissues in the PQ group, WIN 1 mg group and WIN 2 mg group showed diffuse inflammation, which was improved after the intervention of WIN5522-2, especially in the WIN 1 mg group. IL-6 levels of BALF in the PQ group, WIN 1 mg group, WIN 2 mg group and the control group were (1024.77±124.74)U/L, (620.48±99.76)U/L, (823.29±157.88) U/L, and (180.42±20.22)U/L, respectively. IL-6 levels in the WIN 1 mg group and the WIN 2 mg group were statistically lower than those in the PQ group (P=0.021, P=0.016). However, no difference was found between the two intervention groups(P=0.114). The similar condition was also found in TNF-α in BALF and plasma. In the chronic phase, mice in the PQ group, WIN 1 mg group and WIN 2 mg group showed fibrosis in tissue by HE and Masson staining, and the inflammatory condition was improved after the intervention of WIN5522-2, which was more obvious in the WIN 1 mg group. In BALF, TNF-α level was (321.64±50.54)U/L, (260.23±48.19)U/L, (278.89±29.40)U/L, (89.76 ± 10.87)U/L in the PQ group, WIN 1 mg group, WIN 2 mg group and the control group. Differences were found between the WIN 1 mg group and the control group and the WIN 2 mg group. Similar differences were also observed in plasma TNF-α, but not in TGF-β. Conclusions A small dose of WIN55212-2 can improve the general condition of PQ poisoning mice, and reduce the inflammatory and fibrosis-related cytokines levels in PQ poisoning mice.

Roles of human leukocyte antigen-G ( HLA-G) in MDSC proliferation, M1/M2 macrophage differ-entiation and tumor immune evasion / 中华微生物学和免疫学杂志

Objective To investigate the roles of human leukocyte antigen-G ( HLA-G) in mye-loid-derived suppressor cell (MDSC) proliferation and M1/M2 macrophage differentiation in C57BL/6-NCI-H446-G tumor-bearing mice for better understanding the mechanisms of HLA-G involved in tumor immune evasion. Methods NCI-H446 ( human small cell lung cancer cells) and NCI-H446-G ( NCI-H446 cells ex-pressing HLA-G) cells were labeled with CFSE at a final concentration of 1μmol/L. CFSE fluorescence lev-els were measured by flow cytometry at different time points. Mouse tumor models were established by subcu-taneous injection of C57BL/6 mice with NCI-H446 and NCI-H446-G cells, respectively. PBS was used to set up negative control group. The mice in each group were sacrificed to collect tissue samples on 5 d, 10 d, 15 d and 20 d after injection. The percentages of splenic CD11b+Gr1+MDSCs, F4/80+CD80+M1 and F4/80+CD206+M2 macrophages were analyzed by flow cytometry. Results Steady expression of HLA-G in NCI-H446-G cells was confirmed by Western blot and flow cytometry. HLA-G enhanced the proliferation of NCI-H446 cells. Tumor size increased dramatically in tumor-bearing mice in the first five days and then de-creased over time. The tumor-bearing mice in the NCI-H446-G group had larger tumor than those in the NCI-H446 group in every time point (P<0. 05) and required longer time to fully reject the tumor. Compared with the PBS and NCI-H446 groups, the percentage of splenic MDSCs in tumor-bearing mice was significantly in-creased in the NCI-H446-G group (P<0. 05). Moreover, the ratio of M1/M2 in NCI-H446-G tumor-bearing mice was much lower than that in the other two groups (P<0. 05). Conclusion This study indicated that HLA-G could increase the percentage of MDSCs and decrease the ratio of M1/M2, which might illustrate the role of HLA-G in tumor immune evasion and its potential clinical significance in cancer immunotherapy.

The correlation between als3 gene expression and in vivo Candida albicans biofilm formation in mice / 中华传染病杂志

Objective To study the correlation between expression level of als3 gene and the in vivo biofilm formation of Candida albicans in mice.Methods The real-time polymerase chain reaction (PCR) assay was used to detect als3 gene expressions of the clinical Candida albicans isolates from February 2016 to August 2016 in Tianjing No.1 Central Hospital.According to the expression levels of als3 gene, Candida albicans isolates were divided into high and low-expression groups.Thirty C57 mice were randomly assigned to high-expression group (n=15), low-expression group (n=5) and blank group (n=5).Animal model of Candida albicans biofilm was established based on venous catheter and intraperitoneal injection of Candida albicans.Catheters were removed after two weeks;inverted microscope was used for the observation of Candida albicans biofilm formation and transmission electron microscope was used for the observation of its ultrastructure.After irrigating the catheter, the growth of Candida albicans was observed;real-time PCR was used to detect the expression levels of als3 gene 12, 24, and 48 h after the catheter being removed.In this study, t test was used for measurement data and chi-square test was used for rate comparisons.Results In high-expression group, 11 strains (11/15) formed biofilms.In als3 low-expression group, only one strain (1/10) formed biofilm.The difference between these two group was statistically significant (x2=9.64,P0.05).In the als3 high-expression group, the expression of als3 gene declined gradually during the biofilm formation.In the als3 low-expression group, the change of als3 gene expression was not obvious.The expressions of als3 gene over time between two groups were significantly different (t=8.7, 10.3 and 9.2, respectively, all P<0.05).Conclusion The high expression of als3 gene in Candida albicans facilitates the formation of biofilm in vivo.

Construction of Recombinant Adenovirus Vector Expressing Mouse SPINK5 Gene and Its Curative Effect on Skin Lesions in Atopic Dermatitis Mice / 中国医科大学学报

Objective To construct a recombinant adenovirus vector expressing mouse SPINK5 gene,and observe its curative effect on the skin lesions in atopic dermatitis mice model. Methods By recombining DNA technology,the sequence of mouse SPINK5 gene was cloned into adeno?virus shuttle plasmid. Then it was transformed into HEK 293 cells with the adenoviral backbone plasmid to obtain the recombinant adenovirus. A mouse model of atopic dermatitis was established by system and local sensitization of Balb/c mice with ovalbumin . The effect of recombinant adeno?virus on the lesions of atopic dermatitis mice model was observed. Results The SPINK5 over?expressing adenovirus vector and atopic dermatitis mice model were successfully constructed. After 2 weeks of adenovirus?mediated SPINK5 gene intracutaneous injection,the redness and edema of lesions of AD model mice were obvious relieved. The pathological detection indicated that epidermal thickness and prickle cell layer ,inflammatory cell infiltration significant decreased accompanied with the model blank control. Conclusion The adenovirus?mediated SPINK5 gene had signifi?cant therapeutic effect to the atopic dermatitis mice model ,which provided a laboratory basis of application of SPINK5 gene product to therapy atopic dermatitis.

Mechanism of Pharmacological or Toxicity Effect of Ribavirin on Kidney-Yang Deficiency Syndrome Mice Model Infected with Influenza Virus A Based on Serum Metabolomics / 中国药学杂志

OBJECTIVE: To analyze the endogenous metabolite changes in the sera of kidney-yang deficiency syndrome mice infected with influenza virus A after intervention by ribavirin. And to explore the mechanism of pharmacological or toxicity effect of ribavirin. METHODS: KM mice were randomly divided into three groups as normal group, model group and ribavirin group. Mice were infected with virus A after fifteen days Kidney-Yang deficiency syndrome was established. Ribavirin group were orally administrated with ribavirin for 6 consecutive days after inoculation, and the other two groups were given with equal volume of saline solution in the same way. Body weight, rectal temperature were recorded daily. Serum samples were collected from mouse 24 h after the last administration for HPLC-TOF/MS analysis. RESULTS: The results show that ribavirin has good therapeutic effects on the lung index and high mortality rate of mice model. Compared with normal and model groups, the body weight and rectal temperature of them performed falling continuously. The LC-MS data were analyzed with multivariate statistical analysis and 14 potential metabolic markers were obtained which contained D-glucose, sphinganine, linoleic acid and so on. In ribavirin group, metabolism of linoleic acid, arachidonic acid and sphinganine appeared the trend of normal. And sugar and glycerophospholipid became disorders. CONCLUSION: The metabolomics study and pharmacological experiment show that ribavirin might play a role of efficacy in a way that has close correlation with the linoleic acid, arachidonic acid and sphingolipid metabolic pathways. And the toxicity effect may be related to sugar and glycerophospholipid metabolic pathways.

Inhibition of Yuyihe Powder on Tumor Growth in Mice Models of Sarcoma 180 / 中草药·英文版

Objective To explore the antitumor effect of Yuyihe Powder (Yu Yi He San, YYHS) and its antitumor mechanism. Methods After treatment, tumor weight, immune apparatus weight, the life span of transplanted animals, spleen lymphocyte proliferation assays, and IL-2 concentration in mouse serum were recorded or detected. Results YYHS showed strong antitumor ability. Compared with control group, mid-dose YYHS (1.0 g/kg) could inhibit the tumor growth, prolong the life span of S180-bearing mice to some extent, significantly increase the thymic and splenic indices of S180 mice, and strongly promote the secretion of IL-2 in blood; The inhibitory rate on tumor growth and life prolongation rate were 37.1% and 38.37%, respectively. Conclusion YYHS could not only significantly inhibit the growth of S180 cells, but also markedly prolong the survival time of S180 bearing mice. The mechanism of antitumor effect could obviously enhance immunologic function of the S180 bearing mice to inhibit the growth of S180 cells.

Action mechanism of Mahuang Xixin Fuzi decoction for mice with influenza based on metabolomics information / 中国中药杂志

This study aimed to analyze the endogenous metabolite changes in the serum of mice infected with H1N1 virus after intervention by Mahuang-Xixin-Fuzi decoction (MXF) based on metabolomics method, investigate potential biomarkers and related metabolic pathways, and explore the therapeutic mechanism of MXF through metabolomics technology. Thirty-six Kunming (KM) mice were randomly divided into three groups: normal group, model group and MXF group. Influenza virus H1N1 was used by nasal drip to establish influenza mice model. The mice in MXF group were orally administrated with MXF for 6 consecutive days after inoculation, and the other two groups were given with equal volume of saline solution in the same way. Body weight, rectal temperature, morbidity and mortality were recorded daily. Serum samples were collected 24 hours after the last administration for HPLC-TOF-MS analysis. The results showed that as compared with the normal group, the body weight and rectal temperature were decreased in model group, and their lung index and mortality rate were significantly increased (P<0.05); MXF had good therapeutic effects on the abnormity of body weight, rectal temperature, lung index and high mortality rate of mice infected with H1N1 virus. The original data collected from the serum samples were analyzed with R language, MPP, SIMCA-P and other software, and significant changes were found in 14 kinds of endogenous substances from mice serum (P<0.05). As compared with model group, the potential metabolic markers in MXF group recovered to normal levels to a certain degree after being intervened by MXF. Further analysis with MetPA data platform showed that, the pathways involved in 14 metabolites included glucose metabolism, arachidonic acid metabolism, glycerophospholipids and sphingolipids metabolism etc. The metabolomics study and pharmacological experiment showed that MXF might play a role of efficacy by improving glucose metabolism, regulating arachidonic acid metabolism, glycerophospholipid and sphingolipid metabolic pathways.

Effects of Abnormal Phlegmatic Temperament Granules on Behavioristics and Hippocampus Pathomorphism of APP/PS1 Transgenic Mice / 中国中医药信息杂志

Objective To investigate the effects of Abnormal Phlegmatic Temperament Granules on spatial learning and memory, histopathology morphological change in hippocampus CA1 zone; To discuss its mechanism of action.Methods Three-month-old APP/PS1 transgenic mice were randomly divided into 5 groups: model control group, positive control (donepezil 0.92 mg/kg) group, Abnormal Phlegmatic Temperament Granules high-, medium-, and low-dose groups (3, 2, 1.5 g/kg), 18 mice in each group. Another 18 three-month-old C57BL/ 6J mice were chosen as normal control group. All administration groups received relevant medicine for successive 6 months. Then the changes in learning and memory ability of mice were detected by Morris water maze test; pathomorphism in hippocampus CA1 zone was detected by HE staining method; changes of myelin sheath, microtubule, and microfilament in myelinated nerve of hippocampus CA1 zone were detected by electron microscope. Results Morris water maze test results showed that escape incubation period of APP/PS1 transgenic mice was significantly longer than the normal control group (P<0.01), and the original platform time was significantly shorter than normal control group (P<0.01); compared with model control group, Abnormal Phlegmatic Temperament Granules treatment groups escape latency time was significantly reduced (P<0.05, P<0.01). Space experiments and escape incubation period of Abnormal Phlegmatic Temperament Granules high-, medium-, low-dose groups were significantly shortened (P<0.05, P<0.01), and spatial searching test showed that the times of mice in Abnormal Phlegmatic Temperament Granules high-, medium-, low-dose groups passing through effective area increased (P<0.01). The integrity of HE staining pyramidal cell layer in the hippocampus CA1 zones of Abnormal Phlegmatic Temperament Granules high-, medium-, and low-dose groups was relatively good; cells arranged orderly; distribution was normal. Electron microscopic observation showed that compared with model control group, the hippocampus neurons nuclear had irregular shape; nuclear membrane was clear and complete; chromatin was clear; nucleolus was obvious; cell matrix was uniform; organelles were abundant; mitochondrial cristae was obvious; endoplasmic reticulum and free ribosomes were obvious. Conclusion Abnormal Phlegmatic Temperament Granules can improve spatial learning and memory in APP/PS1 mice, alleviate neuronal ultrastructure damage and ultimately improve cognitive function.

Evaluation of Uygur medicine in treatment of tumor by dynamic contrast-enhanced magnetic resonance / 实用放射学杂志

Objective To evaluate whether the functional MRI could be used to reflect the change of angiogenesis after drug treat-ment,and obtain the related semi quantitative and quantitative parameters by dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI).Methods The subcutaneous transplantation colorectal cancer were constructed in 20 nude mice,which were treated with Xiaoaifeimi.The transplanted tumor microvascular density,VEGF and PCNA were monitored.The therapeutic effect and MRI monitoring results were evaluated by combining DCE-MRI with the pharmacokinetic model.The semi quantitative and quantitative parameters were obtained for evaluating the effection of medicine.Half of nude mice were sacrificed to obtain the immunohistochem-ical staining.Correlation between pathological findings and parameters were analyzed.Results Compared to the control group,hu-man colon HT-29 cell proliferation rate was significantly decreased (44.87%)(P <0.05),and the apoptosis rate of HT-29 cells was increased 2.45 times (P <0.05)after Uyghur medicine treatment.The correlation between pathological examination and DCE-MRI parameters showed that Ktrans value and Kep were positively correlated with VEGF,MVD and PCNA (P <0.05).Conclusion A good relationship is showed between immunohistochemistry and DCE-MRI quantitative parameters in Uygur drug treating human colon cancer mice.

Effect of over expression of C-terminal Src kinase binding protein (Cbp) and Cbp palmitoylation to proliferation of T-leukemia Jurkat cell in nude mice / 中国免疫学杂志

Objective:To establish T lineage leukemia Jurkat cell mice model of over expression of C-terminal Src kinase binding protein( Cbp ) and Cbp palmitoylation and to research the effect of Cbp and Cbp palmitoylation to proliferation of Jurkat cell.Methods:Virus transfected cell of neg-EGFP,Cbp-EGFP and Cbp-m-EGFP were used in mice model.24 female BALB/c-nu mice were randomly divided into blank control group,empty virus control group,over expression of CBP group and Cbp palmitoylation group, 6 mice in each group.The nude mice were weighed in 0,1,2,3,4,5 weeks.The amount of white blood cell in peripheral blood were counted in 0, 1, 2, 3, 4, 5 weeks.The proliferation of Jurkat cell in peripheral blood of mice were observed by laser confocal microscope.The pathological changes of liver were observed using HE staining.The proliferation of Jurkat cell in the bone marrow and peripheral blood of mice were detected with flow cytometry.Results:The weight of mice in over expression of Cbp group was less than that in blank control group,but higher than that in empty virus control group and Cbp palmitoylation group.The weight of mice in Cbp palmitoylation group was less than that in blank control group,empty virus control group and over expression of Cbp group.The amount of white blood cell in peripheral blood and proliferation of Jurkat cell in liver, bone marrow and peripheral blood of mice in over expression of Cbp group was higher than that in blank control group, but less than that in empty virus control group and Cbp palmitoylation group.The amount of white blood cell in peripheral blood and proliferation of Jurkat cell in liver, bone marrow and peripheral blood of mice in Cbp palmitoylation group was higher than that in blank control group,empty virus control group and over ex-pression of Cbp group.Conclusion:Over expression of Cbp and Cbp palmitoylation in T lineage leukemia Jurkat cell mice model was established.Over expression of Cbp has inhibitory effect on the proliferation of Jurkat cell.Cbp palmitoylation has promotable effect on the proliferation of Jurkat cell.

Postpartum depression animal model in mice and effect of Yuejuganmaidazaotang on PPD model / 中国药理学通报

Aim To establish a postpartum depression animal model,assess the abnormal maternal behaviors of depressive dams,and observe the rapid antidepres-sant effects of the Yuejuganmaidazaotang (YG)on the PPD model.Methods Thirty-two female Balb /c were randomly assigned to two groups,the control group (Control,con)and the pre-pregnancy stressed group (Vehicle,veh),and vehicle was subjected to 3 weeks chronic restraint stress.After the last stressor,the pre-pregnancy stressed group was housed with a male.Af-ter about 4 weeks later,the mice gave birth to pups. Then at 3 weeks postpartum,we tested the maternal depressive-like behaviors,including sucrose preference test,forced swimming test and novelty suppressed feeding test.Both YG and Ketamine were single ad-ministered 24 hours before behavior test,with single saline for control group and PPD model group.Results After 3 weeks postpartum,the vehicle mice showed depression-like behaviors.Reduced preference in drinking sucrose solution was found in SPT (P <0.01 ).Immobility in FST was significantly increased in vehicle groups (P <0.01 ).In NSFT,the vehicle group displayed a significantly increased latency and reduced unit of food intake compared with control group(P <0.01,P <0.01 ).Acute YG improved per-formance in the SPT(P <0.01),FST (P <0.01)and NSF (P <0.01,P <0.01),which was similar to ket-amine.Conclusions Chronic pre-pregnancy stress can induce dams into postpartum depression.Acute YG exert fast antidepressant effect on this PPD model simi-lar to ketamine.