1.Effect of eszopiclone on pentobarbital sodium-induced sleeping time in acute hypobaric hypoxia mice
Ling ZHONG ; Yongbing SONG ; Jun YANG ; Qian CAI ; Jiangtao XU
Chinese Journal of Behavioral Medicine and Brain Science 2014;23(4):307-309
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.
2.Construction and immunoscreening of cDNA library of Armillifer agkistrodontis nymphs
Ling-ling, ZHANG ; Jia-xu, CHEN ; Shao-hong, CHEN ; Jian, GUO ; Yu-chun, CAI
Chinese Journal of Endemiology 2012;31(6):599-603
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.
3.Effects of acute hypobaric hypoxia on abilities of learning and memory, the level of nitric oxide and endothelin-1 in hippocampus of mice
Ling ZHONG ; Yongbin SONG ; Jianchun XU ; Yan JIAO ; Rong WANG ; Qian CAI ; Jiangtao XU
Chinese Journal of Behavioral Medicine and Brain Science 2014;23(10):878-880
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.
4.Efficacy of ginsenosides combined with prednisone in patients with systemic lupus erythematosus: a prospective, randomized, double-blind, placebo-controlled trial.
Yanli YOU ; Yinglu FENG ; Qing CAI ; Jianlong GUAN ; Lanling ZHANG ; Meijuan XU ; Xia XU ; Changquan LING
Journal of Integrative Medicine 2010;8(8):762-6
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.
5.Effects of pH9.0 Tris-EDTA and pH6.0 citrate on antigen retrieval.
Xin-Lan LUO ; Yan-Hui LIU ; Heng-Guo ZHUANG ; Xiu-Ling CAI ; Jie XU
Chinese Journal of Pathology 2005;34(11):749-751
Antigens
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analysis
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Cervical Intraepithelial Neoplasia
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metabolism
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Citric Acid
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Cyclin-Dependent Kinase Inhibitor p21
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analysis
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Edetic Acid
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Formaldehyde
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Hot Temperature
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Humans
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Hydrogen-Ion Concentration
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Immunohistochemistry
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Intestinal Mucosa
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immunology
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Ki-67 Antigen
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analysis
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Microwaves
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Palatine Tonsil
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metabolism
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Proto-Oncogene Proteins c-bcl-6
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analysis
6.Application of amputation of secondary structures of splenic pedicle and self-made spleen-removing bag in laparoscopic splenectomy using 3-hole method
Guofeng CHEN ; Fuzhen QI ; Dianhua GU ; Jianhuai ZHANG ; Yebo WANG ; Ling LIU ; Yong CAI ; Gang XU
Chinese Journal of Hepatobiliary Surgery 2010;16(9):681-682
Objective To summarize the experience in application of amputation of secondary structures of splenic pedicle and self-made spleen-removing bag in laparoscopic splenectomy the using 3-hole method. Methods The clinical data of 11 patients receiving the procedure from June 2007 to April 2009 in our hospital were retrospectively analyzed. Results Advantages of the procedure were less bleeding, small wound, quick recovery and no occurrence of postoperative complications. Six cases had slight postoperative pain. All 11 patients were cured. Conclusion The technique of amputation of secondary structures of splenic pedicle is safe and feasible in laparoscopic splenectomy using the 3-hole method.
7.Analysis of risk factors for severe acute pancreatitis
Li LI ; Ling HUANG ; Ping XU ; Guijun HE ; Qinghua LI ; Min CAI
Chinese Journal of Pancreatology 2015;15(3):150-153
Objective To investigate the risk factors for prognosis of severe acute pancreatitis (SAP).Methods The clinical data of 563 patients with acute pancreatitis (AP) treated from January 2008 to December 2014 were analyzed retrospectively.There were 334 patients with mild acute pancreatitis (MAP),and 198 patients with moderately severe acute pancreatitis (MSAP),and 31 patients with severe acute pancreatitis (SAP).Risk factors associated with MAP and MSAP + SAP group,MSAP and SAP group were determined by univariate logistic regression,and multivariate analysis was used to determine the risk factors for severity of AP.Results Univariate logistic regression analysis showed that age,WBC,serum glucose,calcium,urea nitrogen arterial partial pressure of oxygen (PaO2),SIRS,pleural effusion,peripancreatic fluid collection were associated with severity of AP in MAP and MSAP + SAP patients;age,serum calcium,PaO2,SIRS,pleural effusion,peripancreatic fluid collection were associated with severity of AP in MSAP and SAP patients.Multivariate logistic regression analysis showed that age,serum calcium,PaO2 and peripancreatic fluid collection were risk factor of MSAP and SAP;age,serum calcimm and SIRS were risk factors of SAP patients for persistent organ failure and pancreatic necrosis.Conclusions For elderly AP patients,measurement of arterial partial pressure of oxygen and serum calcium,peripancreatic fluid collection can help evaluate the severity of AP and early recognition.For SAP patients,the presence of SIRS,high serum urea nitrogen,and low serum calcium and peripancreatic fluid collection raise the concerns of persistent organ failure and pancreatic necrosis.
8.Correlation of toll-like receptor 3 and tumor necrosis factor-α with idiopathic fetal growth restriction
Ling LIU ; Shihong CUI ; Guomei CHENG ; Yajuan XU ; Xiaojuan WANG ; Lindong ZHANG ; Yiming CAI ; Yanni TIAN
Chinese Journal of Obstetrics and Gynecology 2009;44(12):909-914
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.
9.Interferon response-related gene array in predicting the outcome of antiviral treatment in chronic hepatitis C
Tongjing XING ; Hongtao XU ; Wei ZHAO ; Ling SHEN ; Hao LI ; Rentian CAI
Chinese Journal of Clinical Infectious Diseases 2010;03(5):285-289
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.
10.Effects of Chinese herbal medicine Neiyi Recipe-medicated serum on angiopoiesis of endometriosis in the chick chorioallantoic membrane model.
Ling XU ; Qiaoling ZHOU ; Jie HAN ; Danying ZHANG ; Huijuan DONG ; Zailong CAI ; Chaoqin YU ; Dongxia ZHAI
Journal of Integrative Medicine 2012;10(7):800-6
To compare angiopoiesis ability of eutopic and ectopic endometrial tissue isolated from women with endometriosis and endometrium isolated from women without endometriosis (control), and to explore the inhibitory effects of medicated serum of Neiyi Recipe, a compound traditional Chinese herbal medicine.