1.DATA ANALYSIS OF RADIORECEPTOR ASSAY WITH MICROCOMPUTER
Jiang LIU ; Yifan FU ; Xiawen WANG
Journal of Chongqing Medical University 1987;0(01):-
A BASIC program was written to analyse the data from radioreceptor assay by microcomputer. Concentrations of ligand, total binding, nonspecific binding, specific binding and the ratio of specific binding (number of receptor sites) to free ligand content were calculated for each point. The receptor binding affinity constant and binding capacity were obtained by Sca-tchard analysis. Results and graphs can be displayed on the screen and/ or printed out by using a graphic printer
2.The Growth Regulation and Postreceptor Signal Transduction of Somatostatin on Cell Line of Human Colon Cancer
Mingjun SUN ; Ying PANG ; Ruolan JIANG ; Baoyu FU ; Yingjei LIU
Journal of China Medical University 2001;30(1):33-35
Objective:Our purpose was to investigate the growth regulation and postreceptor signal transduction of somatostatin (SS) on human colon cancer cell line. Methods:Low differentiated clone A cells of human carcinoma were treated with different concentrations of SS and the growth status was observed. Intracellular 1,4,5-trisphosphate (IP3) and cyclic adenosine monophosphate (cAMP) were extracted, then the concentrations of them were measured by liquid scintillation technique and gamma scintillation counter.Intracellular free calcium concentration was detected by loading Fura-2 and fluorescental technique. Protein kinase C (PKC) was extracted from cytosol and membrane, then the activity of both parts was determined with TaKai method. Results:The clone A cell growth was inhibited greatly by different concentrations of SS (10-10 to 10-5 mol/L) and it is related to the doses of SS.Somatortatin could largely inhibit the production of intracellular IP3, [Ca2+]i, and cAMP, and decrease the activity of PKC. Conclusion:The growth of Clone A cell can be inhibited by SS. The inhibition may be mediated by phosphate inositol pathway, so intracellular IP3, [Ca2+]i decreased, or inhibited the cell growth by inhibiting the activity of PKC.On the other hand, the cell proliferation may be inhibited by adenosine cyclase pathway, that is decreasing intracellular cAMP ,inhibiting cAMP-depending protein kinase.
3.Screening and identification of HIV-1 gp41 C-helix mimotopes displayed on phages
Beiyi LIU ; Ping ZHU ; Qiangtao HAN ; Shibo JIANG ; Ning FU
Chinese Journal of Immunology 2000;0(09):-
Objective:To find small molecular leads for inhibition on early stage of HIV infection by identification and characterization of the HIV-1 gp41 C-helix mimotopes.Methods:For identification of the gp41 C-helix mimotopes,C7C phage display peptide library was biopanning by using a synthetic peptide N36 which was derived from the gp41 N-helix as target.After three rounds of screening,positive phage clones were identified by ELISA and sequenced.Results:16 of 26 phage clones were identified to bind with peptide N36,and 10 of them were sequenced.Every clone of ten clones contains at least two hydrophobic residues,which may dock into the hydrophobic pocket in the gp41 N-helix domain.9 of the 10 clones have a conservative sequence WW,which may mimic the W628 and W631 in C-helix to interact with the hydrophobic residues in the gp41 pocket.One clone expressing the conservative sequence named clone No.8(CYWWHRLHC) was selected for characterization.The binding between the clone No.8 and N36 was blocked by free peptide N36.And the binding between clone No.8 and peptide N36 was inhibited by peptide C34(IC 50=12.5 ?g/ml).Conclusion:The short circular peptides displayed on phages containing WW residues may mimic the conformational epitope of the HIV-1 gp41 C-helix to interact with the N-helix.This information may be useful for design of HIV-1 fusion inhibitors.
4.Studies on activation,apoptosis and proliferation of NKT like cells in untreated HIV infected patients
Tingting LIU ; Xue ZHAO ; Zining ZHANG ; Yajing FU ; Yongjun JIANG
Chinese Journal of Immunology 2016;32(12):1797-1800
Objective:To study the changes of the NKT like cells after HIV infected. Methods:We collected peripheral blood from 47 untreated HIV infected individuals and 31 healthy controls,and analyzed the expression of Annexin-V,Ki-67,HLA-DR and other surface molecules in NKT like cells by flow cytometry. Results:The NKT like cell percentage of untreated HIV infected group was (3. 03±1. 61)%,the NKT like cell percentage of normal control group was (8. 30±7. 42)%,the percentage of NKT like cells in HIV infected individuals was significantly lower than the healthy controls ( P<0. 05 );the NKT like cell HLA-DR expression of untreated HIV infected group and normal control group were (5. 40±4. 10)% and (0. 89±0. 83)%,the NKT like cell Annexin-V expression of untreated HIV infected group and normal control group were (30. 21±13. 15)% and (5. 40±8. 05)% ,and the activation and apoptosis of NKT like cells was significantly higher after HIV infection compared with health individuals (P<0. 001,P<0. 01),the degree of activation was negatively correlated with CD4 count (r=-0. 885 7,P<0. 05);and the NKT like cell Ki-67 expression of untreated HIV infected group and normal control group were (11. 15±4. 76)% and (27. 63±18. 31)%,the proliferation ability was significantly lower after HIV infection compared with healthy controls(P<0. 05). Conclusion:HIV infection can significantly reduce the number of NKT like cells and its ability to proliferate,and increase its ability to activation and apoptosis.
5.Characterization of the epitopes recognized by monoclonal antibody 3A8 specific to broad-spectrum Gram-negative bacteria
Beiyi LIU ; Xiaotao JIANG ; Haibo LUO ; Ning FU
Chinese Journal of Immunology 1985;0(01):-
Objective:To screen and characterize the common epitopes recognized by monoclonal antibody 3A8,which binds several kinds of Gram negative bacteria,which were recognized by monoclonol antibody 3A8 agaisnt different strains of LPS.Methods:Phage clones binding to 3A8 were screened from C7C phage displayed peptide library,the attached phage clones were eluted by LPS2630(O111:B4).After 3 rounds of panning,the positive phage clones were identified by ELISA,and the amino acid sequences of these positive clones were deduced from DNA sequences.In order to predict how these peptide mimics interaction with 3A8,peptides(A2 and A5) based on conserved sequences of positive clones were synthesized and conjugated with KLH for further study.The binding of A2-KLH and A5-KLH to 3A8 were identified by ELISA.Results:15 of 33 phage clones were identified as positive clones and shown specific binding with 3A8.LPS2630 potently inhibited the binding of phage clone to 3A8.In analysis of the amino acid sequence,there were seven kinds of sequences containing highly hydrophilic residues,and Ser Pro Pro/Pro X Pro was the conserved sequence.The peptides A2 and A5 could bind to 3A8 specially.Conclusion:The conserved sequences containing Ser Pro Pro/Pro X Pro are obtained,which are recognized by mAb 3A8 against broad spectrum Gram negative bacteria and several strain LPS.The synthetic peptides based on the conserved sequence can bind to 3A8,indicating that the peptides can mimic a common epitope of Gram negative bacteria,which be expected as candidate epitope for vaccinization.
6.Study on senescence and proliferation of NKT like cells in HIV infected patients after antiviral therapy
Xue ZHAO ; Tingting LIU ; Zining ZHANG ; Yajing FU ; Yongjun JIANG
Chinese Journal of Immunology 2016;32(9):1354-1356
Objective:To better understand the changes of the NKT like cells after HIV infection and HAART treatment.Methods: Peripheral blood from HIV-infected individuals, HAART-treatment AIDS patients and healthy controls were collected, the expression of CD57 and the proliferation ability of NKT like cells before and after HAART were analyzed by flow cytometry.Results:We found that the percentage of NKT like cells before HAART was significantly lower than the healthy controls ( P<0.01 ) , and recovered after HAART treatment ( P<0.05 );the aging of NKT like cells was significantly higher before HAART compared with health individuals (P<0.01),and recovered after HAART treatment(P<0.05)the proliferation was significantly lower in vitro before HAART compared with healthy controls,and partial recovered after HAART.Conclusion: After HAART treatment,the number of NKT like cells,CD57 expression and the proliferation ability of HIV infected patients were restored.
7.Effect of Trasylol on Expression of CD4~+ and CD8~+ T Lymphocytes in Brain Tissue of Cerebral Ischemia- reperfusion Rats
Chengping JIANG ; Xiaoming WANG ; Bihua WU ; Fu LIU
China Pharmacy 2005;0(21):-
OBJECTIVE: To study protection effect of trasylol on cerebral injury resulting from cerebral ischemia-reperfusion in the interference of immunological response.METHODS: 105 SD rats were randomly assigned into sham operation group,model group and trasylol group (n=35).Focal ischemia-reperfusion model were established by occluding middle cerebral artery.24 h before ischemia and at the 0 h,24 h,4 d,6 d,8 d,10 d,12 d and 14 d of reperfusion,trasylol group were given trasylol via tail vein and other two groups were given normal saline.The infiltration of CD4+ and CD8+ T lymphocytes was observed using immunohistochemistry at different stages of cerebral ischemia-reperfusion.RESULTS: As compared with sham operation group,CD4+and CD8+ T lymphocytes count in model group were increased significantly after cerebral ischemia-reperfusion injury and obtained maximum value at 10th day.CD4+ and CD8+ T lymphocytes count in trasylol group were lower than in model group after cerebral ischemia-reperfusion injury and obtained maximum value at 8th day (P
8.Expressions of heat shock protein (HSP) 60 and 10 in cutaneous squamous cell carcinoma, basal cell carcinoma and solar keratosis tissue
Dongmei LIU ; Yixiu JIANG ; Youla ZHANG ; Chiqing HUANG ; Xuefeng FU
Chinese Journal of Dermatology 2011;44(2):106-109
Objective To measure the expressions of HSP10 and 60 in cutaneous squamous cell carcinoma (SCC), basal cell carcinoma (BCC) and solar keratosis (AK) tissue. Methods Lesion samples were resected from patients with SCC (n = 50), BCC (n = 50) and AK (n = 50), and control samples were obtained from the normal skin adjacent to the operation sites of 14 of the 50 patients with SCC, BCC and AK. Immunohistochemical Envision two step method was used to detect the expression of HSP60 and 10 in the tissue samples.Results The expression of HSP10 was significantly higher in BCC tissue samples (Z = 3.24, P < 0.001 ), but not in AK (Z= 0.74, P> 0.05) or SCC (Z= 0.52, P> 0.05) tissue samples than in the normal control tissue samples. Statistical significance was observed in the expression of HSP10 between AK and SCC and between AK and BCC tissue samples (both P < 0.05), but not between SCC and BCC tissue samples (P > 0.05 ). Elevated expression of HSP60 was found in AK, BCC and SCC tissue samples compared with the control samples (Z =-2.90, -2.15, -2.78,P < 0.01, 0.05 and 0.01, respectively). Furthermore, the expression of HSP60 in SCC tissue samples was higher than that in BCC tissue samples (P < 0.05 ) but similar to that in AK tissue samples. Conclusions There is likely to be a correlation between the high expression of HSP60 and biological behavior of SCC, and between the elevated HSP60 and HSP10 expressions and BCC initiation and development.
9.Study on content of eucalyptol in chao aiye and aiye tan
Liping JIANG ; Guoqian LIU ; Hongxiang XU ; Yinghua FU
Chinese Journal of Biochemical Pharmaceutics 2015;35(10):144-146
Objective To improve the quality of aiye processed products, an eucalyptol content in commercially available aiye two processed products of chao aiye and aiye tan was investigated.Methods A capillary gas chromatography was used.The sample was prepared with n-hexane by reflux condensation.Chromatographic conditions: The separation was carried on an Ailgent DB-1 capillary column(30 mm ×0.320 mm ×0.25 μm). Inlet temperature was 200℃ and FID temperature was 250℃.The programmed column temperature was set as follows:maintained at 100℃ for 6 min and raised to 160℃ at the rate of 20℃/min followed by holding for 3min.The splitting-ratio was 5.0:1.The carried gas was nitrogen, flow rate was 1.0 mL/min.Injection volume was 1μL.Results In the given chromatographic conditions, the eucalyptol chromatographic separation had good, and the separation degree was greater than 1.5 between eucalyptol and other impurity peak.The linear range of eucalyptol was 11.4-114.0 mg/mL(r=0.999 5). Methods repeatability and recovery were good.The minimum limit of quantification was 0.5μg/mL.The results of determination of eucalyptol show that the eucalyptol content in the commercially available 11 batch of chao aiye was between 5.6-78.2 μg /g, and 12 batch of aiye tan had no eucalyptol. Conclusion The processing technology of current commercially available aiye processed products of chao aiye and aiye tan need to be improved, and the quality standard need to be improved.
10.Clinical analysis of 31 cases with AIDS associated oral candidiasis
Qian FU ; Jiang XIAO ; Hongxin ZHAO ; Nan LIU
Journal of Practical Stomatology 2014;(6):839-841
Objective:To study the clinical features and treatment outcome of AIDS associated oral candidiasis.Methods:The clinical data of 31 cases with AIDSassociated oral candidiasis from 201209 to 201303 were studied retrospectively,including general data,clinical features,oral manifestation,CD4 cell count,opportunistic infections,and antifungal therapy outcome,etc.Results:CD4cell count <200 cell/μl was found in 30 cases,AIDSrelated multiple opportunistic infection was observed in 29 cases.30 cases hadpseudomembranous candidiasis,1 cases had erythematous candidiasis and 2 cases had pseudomembranous candidiasis with angular candidiasis.After antifungal treatment,the lesion of 8 cases reduced,that of 23 cases disappeared completely,lesion relapse after drugwithdrawal happened in 3 cases.Conclusion:AIDSassociated oral candidiasis was more common in AIDS patients with CD4 <200cells/μl,the main clinical form is pseudomembranous type,and with multiple opportunistic infections.The antifungal treatment is effective for the patients.