1.Role of sphingolipids signaling in pathogenesis of pulmonary fibrosis
Jiping YI ; Ming ZENG ; Xingxuan HE
Chinese Journal of Pharmacology and Toxicology 2016;(2):158-164
Pulmonary fibrosis is a group of chronic lung diseases caused by various factors and characterized by chronic inflammations,lung tissue structure damage,increase of pulmonary interstitial collagen and massive deposition of extracellular matrix (ECM). Because of its complicated etiology, there is no effective treatment currently. Recent studies showed that the activation of sphingolipids signaling and pulmonary fibrosis were closely related. This paper describes the composition and function of sphingolipids signaling pathway and its effect on fibrosis in order to provide new ideas about further study of the pathogenesis of pulmonary fibrosis and methods of prevention.
2.Experimental Model of Acute Myocardial Infarction Induced by Ligation of Coronary Artery in Rabbits
Lin SUN ; Yi LI ; Xuefeng GUANG ; Zhang XU ; He ZENG
Journal of Kunming Medical University 2001;22(1):39-41
Objective an experimental animal model of acute myoc ardial infarction (ANI) was established by opening chest and ligation of the left anterior descending (LAD) coronary artery. Methods a total of 20 rabbits were opened chest and ligated LAD under sterilization. Electrocardiogram (ECG) and myocardial-enzymes in blood serum were investigated. Results ECG of all rabbi t s showed normal before operation. Irmediately after and 1/2 hour after ligation , ST-segment elevated and ECG showed ambulatory changes for 7 and 9 rabbits respectively. Two hours after LAD ligation, the change of ECG for 2 rabbits wa s not typical and 2 of them died during experiment. Four weeks after operation, E CG of 18 rabbits showed the chest leads had pathologic Q waves. Twenty-four ho urs after LAD ligation, AST, LDH, LDH-1, CK and CK-MB in blood serum were significantly increased. There was significant difference compared with before operation (except LDH) (P<0. 0l). Conclusions:The method was sim ple and well repeated. The formation of myocardial infarction was reliable and rabbits were maintained for a long time after operation. It provides a valuable animal mode l for the experiment study of coronary heart disease.
3.In vitro antifungal activity of tacrolimus alone or in combination with itraconazole or terbinafine against Exophiala dermatitidis
Chengyan HE ; Yi SUN ; Lujuan GAO ; Ming LI ; Tongxiang ZENG
Chinese Journal of Dermatology 2017;50(4):283-285
Objective To evaluate in vitro antifungal activity of tacrolimus combined with itraconazole or terbinafine against Exophiala dermatitidis (E.dermatitidis).Methods The minimum inhibitory concentrations (MICs) of itraconazole and terbinafine against 12 strains of E.dermatitidis were determined using the Clinical and Laboratory Standards Institute (CLSI) broth microdilution susceptibility method (M38-A2 Document).A broth microdilution checkerboard method was used to evaluate the in vitro antifungal activity of tacrolimus combined with itraconazole or terbinafine against E.dermatitidis.Results The MIC ranges of terbinafine and itraconazole against E.dermatitidis were 0.060-0.125 mg/L and 0.5-1 mg/L,respectively.The combination of tacrolimus with terbinafine showed synergistic inhibitory effects against 5 strains of E.dermatitidis,while the combination of tacrolimus with itraconazole revealed synergistic effects against 10 strains of E.dermatitidis.No antagonism was observed in either of the two combinations.Conclusion In vitro combination of tacrolimus with itraconazole or terbinafine can enhance the antifungal activity of itraconazole or terbinafine against E.dermatitidis.
4.Age-related changes in phenotypes of T lymphocytes in human peripheral blood
Xianhui HE ; Lihui XU ; Yi LIU ; Yaoyin ZENG
Chinese Journal of Pathophysiology 2000;0(08):-
AIM: To investigate the details of age-related changes of T lymphocytes in order to seek for sensitive biomarker for immunosenesence. METHODS: Heparin anticoagulated venous blood was collected freshly from young (20-35 years) and elderly (50-75 years) volunteers and three or four color immunofluorescence staining was performed. The nucleated cells were acquired and the phenotypes of T lymphocyte subpopulations were analyzed with flow cytometry. RESULTS: There was no significant difference in percentages of pan-T (CD3 +), helper T (CD4 +) and cytotoxic (CD8 +) T subsets between young and elderly, whereas the density of CD3 molecule (MFI) on T cells in elderly group decreased significantly. It was also found that the rates of CD44 + and CD62L + T cell subsets in young group did not have statistical difference from elderly. However,the rates of CD95 + pan-T, helper T and cytotoxic T subsets of elderly group were all markedly higher than that in young group. CONCLUSIONS: The relative rates of T cell and its subsets displayed no age-related changes while the density of CD3 was down-regulated during aging in these groups investigated. Moreover, the expression percentage of CD95 (Fas) on T cells increased as aging, suggesting that it is a potential biomarker for evaluating immunosenescence.
5.Effects of Ghrelin on human alveolar epithelial cell apoptosis in response to lipopolysaccharide through AKT/ERK signaling pathway
Chunrong HUANG ; Wanmei HE ; Hui YI ; Haichong ZHENG ; Mian ZENG
Chinese Journal of Immunology 2017;33(8):1129-1134
Objective:To investigate the protective effects of Ghrelin on LPS-induced apoptosis of human alveolar epithelial A549 cells,along with the possible molecular mechanisms.Methods: CCK-8 assay was used to examine the cell viability of A549 treated by LPS.Apoptosis of A549 cells was measured by TUNEL.NO(Nitric oxide)production was detected by NO-specific fluorescent probe 3-Amino,4-aminomethyl-2′,7′-difluoresceindiacetate(DAF-FM DA).Western blot was also performed to examine the expressions of iNOS(inducible nitric oxide synthase),AKT,ERK,p-AKT,p-ERK and apoptotic proteins,such as Bcl-2,Bax,and cleaved caspase-3.Results: LPS exposure impaired cell viability and increased apoptosis of A549 cells significantly in concentration-and time-dependent manners accompanied with increased Bax and cleaved caspase-3 production,coupled with decreased Bcl-2 levels.Meanwhile,LPS promoted iNOS expression and the production of NO.Ghrelin pretreatment ameliorated LPS-caused alterations in the ratio of Bax/Bcl-2 and cleaved caspase-3 expression.TUNEL analysis showed that Ghrelin could decrease the apoptosis induced by LPS in A549(P<0.05).Simultaneously,LPS remarkably decreased the expression of p-AKT and p-ERK in A549 cells,which was abrogated by Ghrelin pretreatment.However,Ghrelin had no significant effect on NO production induced by LPS.Conclusion: Ghrelin reduces LPS-induced apoptosis of human alveolar epithelial cells partly through activating the AKT and ERK pathway,but the level of iNOS derived NO could not be reduced.
6.PRELIMINARY STUDY ON SCREENING OF PHAGE RANDOM PEPTIDE LIBRARY USING RABBIT SERUM FROM A NEW MODEL INFECTED WITH SCHISTOSOMA JAPONICUM
Yilan HU ; Li HE ; Mingsen JIANG ; Xinyuan YI ; Xianfan ZENG
Chinese Journal of Schistosomiasis Control 1989;0(02):-
Objective To screen the 12 mers-phage random peptide library using the serum from the new model rabbit and to identify the immuno-protection of the positive phages. The new model infected with Schistosoma japonicum was proved that has a high protection against the challenge infection. Methods After being absorbed by E.coli antibody, the serum of the new model rabbit was used to screen the peptide library. Through three rounds of biopaning and enriching, lots of positive phages were obtained and their antigenic ability was tested. Every mouse was immunized by subcutaneously injecting 1?10 14 pfu positive phages from the new model rabbit serum respectively at 0-2-4 th week. After 4 weeks of the last immunization, the challenge infection was performed. At the same time, several control groups including the group immunized with the phages from the rabbit serum of the normal model infected with Schistosoma japonicum, the group immunized with the original 12 mers-phage random peptide library and the control group of challenge infection were arranged. Results ①The positive clones of phage(1?10 14) from the new model rabbit serum were strongly recognized by the rabbit serum of the new model, weakly recognized by the rabbit serum of the normal model infected with Schistosoma japonicum,but not recognized by the serum of healthy rabbit. ②The reduction rate of adult worms and liver eggs induced by phages screened with the rabbit serum of the new model group and the nomal model group and that induced by the original peptide library were respectively 27 2% and 38 8 %, 17 8% and 35 0%, 4 5% and 6 0% Conclusion The new model group obtained a higher reduction rate of adult worms than the nomal model group (P
7.Construction of mammalian cell expression vector for HLA-A~*0201 and EGFP fusion protein and its expression and localization in K562 cells
Xianhui HE ; Lihui XU ; Yi LIU ; Xiaochang CAI ; Yaoying ZENG
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To construct the mammalian cell expression vector for enhanced green fluorescent protein (EGFP) and HLA-A*0201 fusion protein and analyze its expression and subcellular localization in the transfected K562 cells. METHODS: The HLA-A*0201 cDNA was cloned by RT-PCR and the gene was inserted into pEGFP-N1 to construct a vector for the fusion protein. The expression of the fusion protein in K562 cells transfected with the vector was evaluated by flow cytometry and its subcellular localization was investigated by confocal microscopy. RESULTS: The full-length encoding region of HLA-A*0201 cDNA was cloned from two HLA-A2 positive donors and the expression vector for the HLA-A*0201-EGFP fusion protein was constructed by PCR using a primer pair to introduce a Kozak sequence before ATG and the stop codon was deleted. Five hours after K562 cells was transfected with the vector, the expression percentages of HLA-A*0201 and EGFP were 25.12?2.26 and 27.37?3.59, respectively and no significant increase was observed after 24 h. The fusion protein was predominantly located on the membrane with low level distribution within the cells. In contrast, no HLA-A*0201 but only EGFP was detected in the empty vector transfected K562 cells and the EGFP was dispersed within the cells. CONCLUSIONS: The expression vector for HLA-A*0201-EGFP fusion protein was constructed and the fusion protein expressed in K562 cells was primarily distributed on the membrane. The results suggest that the transfected K562 cells are potential antigen-presenting cells.
8.Significance of Monitoring Serum Level of Gentamycin in Children with Serious Illness
ping, ZENG ; yi-yu, YANG ; jian-ping, TAO ; yan-ling, HE ; yi-lu, CHEN ; qi-yi, ZENG
Journal of Applied Clinical Pediatrics 1992;0(06):-
Objective To explore the serum level of gentamycin for orally in children with serious illness.Methods The serum level of gentamycin in 41 children who were in serious illness [multiple organ dysfunction(MODS)group with 21 cases and non-MODS group with 20 cases ] were monitored and the patients were treated with select decontamination of the digestive tract(SDD) from October 2004 to April 2005.Dosage:10 mg/(kg?d),orally taken three times(every 8 hours) one day.The blood after taking the drug one hour later in the fourth day was selected and the serum level of getamycin was monitored.Results Thirty-six children of 41 cases serum level of gentamycin were negative and 5 children(4 in MODS group and 1 in non-MODS group) who had alimentary tract hemorrhage were masccline in serum after taking gentamycin one hour later in the forth day.The absorption of gentamycin from enteric after orally was not(rela)-ted to MODS.There were statistics value between the gestrintestinal tract ulcer and serum level of gentamycin.Conclusions The safety for treating the children in serious illness with gentamycin for SDD is obvious.But we suggest to monitor the serum level of gentamycin for who has severe alimentary tract hemorrhage together with insufficiency of liver and kindey.
9.Serum immunopharmacological assessment:effects of cycloheximide on the activation of T lymphocytes of mouse
Shiming QUAN ; Yaoying ZENG ; Xianhui HE ; Shan ZENG ; Jingfang DI ; Jingxian ZHAO ; Yi LING
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To investigate the effect of cycloheximide on the T cells activation by mitogen in vitro with CD69 expression as activation marker for the application of this drug clinically. METHODS:Lymphocytes were isolated from lymphoid nodes of C57BL/6 mouse. The cells were preincubated with cycloheximide(CHX), 5% serum containing CHX respectively for an hour, then further incubated with polyclonal activators (Con A or PDB). Harvesting the cells after whole incubation for 24 h, we estimated the expression rates of CD69 on T cells by flow cytometry following two-color immunofluorescent staining. RESULTS: The expression rates of CD69 on the T cells preincubated with CHX, serum containing CHX after the stimulation in response to Con A or PDB all showed significant difference with the expression rates of control group, respectively ( P
10.The effect of recipient kidney function by Ber coadministration CsA used to induce immune tolerance in rats of allogenic cardiac transplantation
Xinghua NI ; Yaoying ZENG ; Jingfang DI ; Xianhui HE ; Tongcheng LIU ; Yi LIN ; Wenyue XIE ; Shan ZENG ; Zheng FENG ;
Chinese Pharmacological Bulletin 2003;0(10):-
AIM To investigate the effect of recipient kidney function by CsA coadministration Ber used to induce immune tolerance in rats of allogenic cardiac transplantation. METHODS The authors established the SD to Wistar rats heterotopic cardiac transplantation model by Onos methods.Observe the cardiac allograft survival and levels of BUN and Cr in the recipients plasma. The recipients were classified into 5 groups randomly after heterotopic cardiac transplantation were performed. Group A (Wistar to Wistar)): Received placebo intraperitoneal injected for 21 days; Group B (SD to Wistar): Saline intraperitoneal injected for 21 days; Group C (SD to Wistar):CsA 2 mg?kg -1 ?d -1 intraperitoneal injected for 21 days; Group D(SD to Wistar):Ber 16 mg?kg -1 ?d -1 gastrointubation for 21 days; Group E(SD to Wistar): Ber 16 mg?kg -1 ?d -1 gastrointubation coadministration CsA 2 mg?kg -1 ?d -1 ip for 21 days. RESULTS The levels of BUN and Cr in recipint plasma is lower evidently compare with the group with CsA ip simply. CONCLUSION Ber can reduce the renal toxicity in recipients by CsA which was intraperitoneal injected (ip) over a long period time.