Objective To assess the effects of eszopiclone (ESZ) on the pentobarbital sodium-induced sleeping time and spontaneous activity in mice exposed to acute hypobaric hypoxia.Methods 120 mice were randomly divided into 6 groups by using two factors 2×3 levels factorial design,in which two factors were interventions (ESZ and 0.9％ sodium chloride,2 levels) and altitudes (800 m,3500 m and 6000 m,3 levels).The pentobarbital sodium-induced sleeping test and the open field test were engaged to assess the effects of ESZ on sleeping time and spontaneous activity.Results (1) The drug and altitude had no interaction in the results of both the pentobarbital sodium-induced sleeping test and the open field test(P＞0.05).(2)The time of pentobarbital sodium-induced sleeping of mice in the groups of ESZ at each altitudes were (37.77± 18.22) min,(37.02± 13.67) min,(95.67±47.68)min and in the groups of NS were(17.78± 14.10) min,(15.09± 12.46) min,(39.54±28.24) min respectively,and the sleep time in ESZ groups were significantly longer than those in the groups of NS (P＜0.05).The time of pentobarbital sodium-induced sleeping were longer in group of 6000 m than those in the other two groups,both the ESZ and NS groups (P＜0.05).(3)No significant difference was found in the open field test between the ESZ and NS groups in the same altitude(P＞0.05) ; while the mice at the altitude of 6000 m in groups of ESZ and NS decreased compared with the groups at the altitude of 800 m after the relevant drugs intra-perineally for 6 h (P＜0.05).Conclusion ESZ may prolong pentobarbital sodium-induced sleeping time especially at the altitude of 6000 m and with no influence on the spontaneous activity in mice exposed to acute hypobaric hypoxia.High altitude at 6000 m may prolong the sleep time induced by pentobarbital sodium and reduce the spontaneous activities.

Objective To construct cDNA entry library and cDNA expression library of Armillifer agkistrodontis (A.) nymphs and make a preliminary immunoscreening for the cDNA expression library.Methods The nymphs were collected from the Kunming mice infected experimentally with A.agkistrodontis eggs and the total RNA were extracted from the nymphs using TRIzol Reagent.After purifying the mRNA,the synthesized cDNAs were cloned into the donor vector pDONR222 by BP reaction of Gateway technology and the recombinants were transformed into the DH10B cells by electroporation,the cDNA entry library was obtained.Next,the expression vector pDEST17 was ligated with entry clones by LR reaction,and the recombinants were transformed into the BL21 (DE3) cells.Hence,the cDNA expression library was constructed.Then,the expression library was immunoscreened with the mixed sera of mice infected with A.agkistrodontis,and the insertions of positive clones were sequenced.After that,the open reading frame(ORF) of positive slone sequence,the homology of the screened genes and their encoded proteins were analyzed by Finder and BLAST (basic local alignment search tool) program of National Center of Biotechnology Information(NCBI),and the discovered new genes were submitted into the GenBank.Besides,the physico-chemical properties,secondary structure and B cell epitopes of encoded proteins were also analyzed by bioinformatics software.Results The average titer and total clones of the cDNA entry library were 1.45 × 105 CFU/ml(colony-forming unit,CFU) and 1.74 × 106 CFU,respectively,and the range of fragment length of the inserted cDNA was between 0.2-4.0 kb,with an average of 1.4 kb.The total clones of cDNA expression library were 1.00 × 105 CFU,and the fragment length of the inserted cDNA was between 0.3-2.2 kb,with an average of 1.0 kb.Five positive clones,coded S1,S5,A1,D1 and F1,respectively,were obtained through preliminary immunoscreening.The sequence and homology of the five positive clones were sequenced and analyzed by BLAST program.No significant similarities were found in pentastomida species,which meant that they were all novel genes of A.agkistrodontis.The gene sequences were submitted to GenBank,with the accession number from JQ180451 to JQ180455.Also,results obtained by bioinformatics software showed that the predictive encoding proteins were all potential to be valuable recombinant diagnostic antigens.Conclusions The cDNA library of A.agkistrodontis nymphs is successfully constructed,and five new genes of A.agkistrodontis are discovered.The establishment of cDNA library and the discovery of the new genes will lay a foundation for further studying the gene functions and screening the immunodiagnostic antigens.

Background: The side effects of glucocorticoid in treatment of systemic lupus erythematosus (SLE) have been the focus of debate, and our preliminary study indicates that ginsenosides can enhance the efficacy of dexamethasone. Objective: To observe the effects of ginsenosides combined with prednisone in SLE patients. Design, setting, participants and interventions: A total of 60 SLE patients from Department of Rheumatology and Immunology, Changhai Hospital, Second Military Medical University, were randomly divided into treatment group and control group, with 30 patients in each group. Patients in the treatment group were given routine treatment with prednisone plus ginsenosides, while those in the control group were given routine treatment with prednisone plus placebo. They were all treated for 3 months. Main outcome measures: After three-month treatment, syndrome score in traditional Chinese medicine (TCM), total response rate and symptom improvement rate were measured and evaluated. Results: Twenty-eight cases in treatment group and twenty-seven cases in control group were included in analysis. The total response rates in the treatment group and control group were 89.28% and 66.67% respectively, and there was a significant difference between the two groups (P<0.05). After treatment, the TCM syndrome scores in the two groups were lower than those before treatment (P<0.01), and prednisone plus ginsenosides was better in decreasing the TCM syndrome score than prednisone plus placebo (P<0.05). The symptoms were improved in the treatment group as compared with the control group (P<0.05). Conclusion: Prednisone combined with ginsenosides can increase the clinical effective rate and improve the clinical symptoms of SLE patients.

Objective To examine the effects of acute hypobaric hypoxia on abilities of learning and memory,the water content in brain,the level of nitric oxide(NO) and endothelin-1 (ET-1) in hippocampus of mice.Methods The acute hypobaric hypoxia environment were made by putting the mice in a hypobaric chamber simulated at altitude of 6 000 meters for 8 hours.The capabilities of learning and memory of mice were detected by Morris water maze test.The content of water in hippocampus were examined by measuring the ratio of dry/wet weight,and the level of NO and ET-1 in brain was detected by colorimetric method.Results Morris water maze test showed that the mean escape latency of mice in the hypobaric hypoxia group were longer than that in the normobaric normoxia group((44.60±7.80) s vs (26.39±8.44)s,P＜0.01),and the target quadrant residence time were decreased((19.78±2.74) s vs (22.98±6.14)s,P＜ 0.05).Compared with the normobaric normoxia group (NO:(2.37 ± 1.07) μmol/gProt,ET-1:(38.87 ± 6.17) ng/L),the water content in brain of mice in the hypobaric hypoxia group was increased (P＜ 0.05),meanwhile,both the level of NO ((4.48 ± 1.45) μmol/gProt) and ET-1 ((52.09±6.75)ng/L) in brain were increased too(P＜0.01).Conclusion Hypobaric hypoxia can decrease the abilities of learning and memory of mice,and these changes might be related with the increased water content and the increased level of NO and ET-1 in hippocampus of mice.

Antigens ; analysis ; Cervical Intraepithelial Neoplasia ; metabolism ; Citric Acid ; Cyclin-Dependent Kinase Inhibitor p21 ; analysis ; Edetic Acid ; Formaldehyde ; Hot Temperature ; Humans ; Hydrogen-Ion Concentration ; Immunohistochemistry ; Intestinal Mucosa ; immunology ; Ki-67 Antigen ; analysis ; Microwaves ; Palatine Tonsil ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; analysis

AIM:To evaluate the application and effect of bandage contact lens in pterygium excision combined with conjunctival transplantation (CAT).METHODS:In a prospective, randomized, controlled clinical study, 110 patients (110 eyes) diagnosed with primary pterygium were collected in PLA NO.474 Hospital from January 2015 to January 2016.The 110 patients enrolled in the study in turn, and divided into two groups by the odd and even number.The odd number divided into bandage contact lens group (CAT + bandage contact lens, n=55), while the even number divided into control group (CAT, n=55).Visual analog scale (VAS) and corneal irritation were evaluated on 1, 3 and 7d after operation.Cornea fluorescent staining testing was carried out on 3d after operation.Following-up all the patients with 1a at least observed the recurrence of pterygium.RESULTS:The score of VAS in bandage contact lens group less than that in control group on 1d (4.13±2.06 vs 5.80±1.93, t=4.391, P<0.001) and 3d (2.09±1.36 vs 3.65±1.65, t=5.422, P<0.001) after operation, while there was no significant difference between two groups on 7d (t=1.295, P=0.198) after operation.The corneal irritation in bandage contact lens group less than that in control group on 1d and 3d after operation (P<0.05), while there was no significant difference between two groups on 7d after operation (P=0.052).Cornea fluorescent staining testing area in bandage contact lens group was less than that in control group on 7d after operation (0.33±0.37mm2 vs 2.73±2.21mm2, t=7.921, P<0.01).There was no significant difference in recurrence rate between two groups after 1a operation (P=1.000).CONCLUSION:Bandage contact lens could significantly release pain and corneal irritation, promote the healing of the corneal epitheliums in the defected area, and increase the postoperative comfort level in patients after operation.

Objective To study the role of hematopoietic growth factor(HGF)of placenta chorionic villus in fetal hematopoiesis during embryo ontogeny by observation of the appearance time and the content changes with the fetal growth, which was compared with HGF in cord blood. Methods Thirty embryo villus (2 g each) and 30 cord blood (2 mL each) were collected separately from early pregnant stage(6- 8 weeks), middle pregnant stage(16-22 weeks)and late pregnant stage (37-42 weeks). The levels of HGF were detected by enzyme - linked immunosorbent assay. Results HGF were produced on the early pregnant stage and the content of FL-T3,IL-3 increased gradually.There were significantly differences at different stages(P

To study the cognitive effects of diterpene ginkgolides (DG), transient middle cerebral artery occlusion (tMCAO)-induced rats were established. tMCAO-rats induced by suture method were divided into sham operation group, solvent control group, NBP group, DG group. The animal experiments in the present study were performed in accordance with the Ethical Guidelines of the Laboratory Animal Welfare Ethical Committee of Peking Union Medical College (00000646, 00000635). The effects of DG on tMCAO rats were evaluated by neurological severity score, cerebral infarction volume measurement, step-down and Morris water maze test. In the acute tMCAO rat model, 100 mg·kg^-1 DG improved the neural score and infarction volume. In the chronic tMCAO rat model, DG 100 mg·kg^-1 significantly improved the survival rate of tMCAO-induced rats. The Morris water maze results showed 100 mg·kg^-1 DG decreased the latency of tMCAO-induced rats to find the platform, while the effect was weaker than the NBP. However, DG 30 mg·kg^-1 did not show a significant effect. In conclusion, DG exerted a therapeutic effect on transient middle cerebral artery occlusion.

Objective To investigate the expression and the significance of toll-like receptor 3 (TLR-3)in placenta,tumor necrosis factor-α(TNF-α)in maternal and cord blood of idiopathic fetal growth restriction(IFGR),and their correlation with the pathogenesis of symmetric and asymmetric IFGR.Methods From April 2008 to April 2009,42 primiparae of singleton pregnancy and their IFGR babies,who delivered at term through cesarean section, in the Third Affiliated Hospital of Zhengzhou University were enrolled. All subjectects were divided into symmetric IFGR group (n=20) and asymmetric IFGR group (n =22). Another 42 non-IFGR pairs were randomly selected as the control group. The polink-2 plus polymerized horseradish peroxidase (HRP) immunohistochemical method and the enzyme linked immunosorbent assay (ELISA) were applied to detect TLR-3 and TNF-α levels. Results (1) The expression of TLR-3 protein were observed in all maternal placenta of the three groups. TLR-3 essentially expressed in syncytiotrophoblasts and hofbouer cells in the symmetric IFGR and control group, but expressed mostly in hofbouer cells and less in syneytiotrophoblasts in the asymmetric IFGR group. (2) The expression of TLR-3 in the syncytiotrophoblasts of the symmetric and asymmetric IFGR group was significantly lower than in the control group (111±14 and 118±11 vs. 156 ± 9, P<0. 01). The number of TLR-3 positive in Hofbourer cell in the symmetric IFGR group was lower than the control group (8. 9±2. 8 vs 17.5±2. 8, P <0. 01 ), but the number in the asymmetric IFGR group was higher (23.8±3.7) compared with the control group (P <0. 01). (3) The TNF-α levels in the maternal and cord blood of the symmetric and the asymmetric group were higher than that of the control group [maternal : (90±10) μg/L and ( 86±11 ) μg/L vs. (73±9) μg/L;cord blood: (92±12) μg/L and (96±8) μg/L vs. (79±9) μg/L;P<0.01]. (4) Neither symmetric nor the asymmetric IFGR group showed any correlations between the maternal and cord blood levels of TNF-α (P>0. 05). (5) Significant correlation was found between the TNF-α level of the cord blood and TLR-3 expression in the placenta in both the symmetric and asymmetric IFGR group(P<0. 05),but no relationship was found between the maternal blood TNF-α level and TLR-3 expression in the placenta (P>0. 05). Conclusions The variantions of TLR-3 expression in placenta and the increased expression of TNF-α in cord blood are associated with the genesis IFGR. The reduced expression of TLR-3 may related to symmetric IFGR, while the increased TLR-3 level in hofbouer cells may lead to asymmetric IFGR.

Objective To explore the application of interferon(IFN)response-related genes in predicting the outcome of antiviral treatment in chronic hepatitis C. Methods SuperArray microarray was used to detect the expression of IFN response-related genes in peripheral blood monocytes(PBMC)from chronic hepatitis C patients before treatment. SPSS 12.0 was used for statistical analysis. Results Ten patients were classified as rapid responders(RVR)and seven patients as non-RVRs according to the serum HCV RNA level after 4 weeks of treatment in 17 patients. Compared with healthy controls, nine differentially expressed genes were found in RVR patients, one up-regulated and eight down-regulated; eighteen differentially expressed genes were found in N-RVR patients, all down-regulated. Five differentially expressed genes were found between the patients with RVR and N-RVR: four up-regulated genes were PRKCZ, PRKRA, IRF5 and TNFSF10(t =5.44, 3.13, 5.24 and 2.30, P=0. 000, 0.010, 0.005 and 0. 044); one up-regulated gene was IFIT5(t = 2.43, P = 0. 035). Of seventeen patients, 12 were HCV genotype 1b, 5 were HCV genotype 2a. Compared with HCV2a, IFI6 and IFI44 gene in HCV1b were downregulated(t = 2.42 and 2.45, P = 0. 038 and 0. 033). Conclusions The expression of IFN responserelated genes is associated with response to IFN treatment. HCV genotype 1 b is more successful in inducing the down-regulation of IFN response-related genes than that of HCV genotype 2a, thus leading to the resistance to IFN.