1.Experimental study on influence of cytarabine on K562 cells proliferation and apoptosis by autophagy pathway
Hao LUO ; Zan MENG ; Zehong LIU ; Xiaolu CHEN
Chongqing Medicine 2017;46(13):1736-1739
Objective To investigate the effect of cytarabine (Ara-C) on proliferation and apoptosis of human erythroleukemia K562 cell linethrough autophagy pathway and its possible mechanism.Methods The cellular proliferation inhibiting rate after different concentrations of Ara-C acting for 24,48 h was detected by CCK-8;the cell cycle and apoptosis were detected by flow cy tometry(FCM);the chromatin morphological changes in nucleus were observed by Hoechst staining;the cell acidic autophagy vesicles were detected by acridine orange staining;the expression changes of p38 and p-p38 proteins were detected by Western blot.The expressions of autophagy apoptosis related gene and protein were examined by RT-PCR and immunofluorescence.Results The CCK-8 results found that different concentrations of Ara-C could inhibit the proliferation of K562 cells with dose-and time-dependent manners.FCM detecting indicated that Ara-C could increase apoptosis and could arrest the cell cycle at S phase;Hoechest staining showed that K562cells had typical apoptotic morphological changes after Ara-C treating;the Acridine orange staining revealed that Ara-C caused the inclease of the green fluorescene in cells of the Ara-C group,and the cells appeared a great number of acidic autophagy vesicles;RT-PCR results showed that Ara-C up-regulated the expression of autophagy key genes Beclin-1,LC3A and LC3B;Western blot results showed that Ara-C increased the expression of phosphorylated p-p38.Immunofluorescence results showed the expression of LC3B was significantly enhanced.Conclusion Ara-C canactivate p-p38 mediated K562 cells to generate autophagy,then inhibit the cell proliferation and promotes apoptosis.
2.Antigenic Localization of Specific Allergen in the Body of Dermatophagoides pteronyssinus by Immunohistochemistry
Renlong FU ; Zhigang LIU ; Miao XING ; Li LI ; Zehong ZOU
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(04):-
Objective To study the localization of specific allergen of Dermatophagoides pteronyssinus. \ Methods\ Through optical microscope,the specific allergens of D.pteronyssinus were observed in paraffin sections using D.pteronyssinus\|specific IgE antibodies from the patient sera. \ Results and Conclusion \ The digestive system was found occupying large parts of body cavity of D.pteronyssinus by HE staining, while the specific allergens of D.pteronyssinus were mostly occurred in the midgut tissue, gut contents, cuticle and reproductive system in the immunostained sections. The results also showed that many parts of D. pteronyssinus were recognized by the specific IgE antibodies obtained from allergic individuals to D.pteronyssinus, which provided a theoretic base for further study of isolation and purification of the specific allergen.
3.Allergenicity evaluation and three dimensional structure analysis of pollen panallergen Profilin from Amaranthus spinosus L.
Ailin TAG ; Linchuan LIU ; Yongfei WANG ; Zehong ZOU ; Sanmei MA ; He LAI ; Lu YU ; Qiurong WU
Chinese Journal of Microbiology and Immunology 2008;28(7):616-620
Objective To clone and characterize Profilin encoding genes in Amaranthus spinosus and to analyze the contribution of different amino acids in isoallergens to allergen antigenicity and tertiary structure. Methods The primers were designed according to the core sequences which were obtained by bioinformatic analysis of the known Profilin amino acid sequences, followed by gene cloning from the Ama- ranthus spinosus cDNA pool and subsequent confirmation by double-digestion, colony PCR and DNA sequen- cing. Antigenicity evaluation and tertiary structural modeling of the encoded protein were accomplished by online software MULTIPRED and SWISS-MODEL, respectively. Results Two panallergenic genes, named as PRF7 and PRF23, were acquired from Amaranthus spinosus. Sequence and structure analysis demonstra- ted that there was some discrepancy in tertiary structures of the encoded proteins, besides distinct difference in their amino acid sequences. PRF7 exhibited high homology with panallergen Profilins Q64LH0, with the identities 98%, whereas the homology of PRF23 and Q9XF42 (apple allergen) was 81%. Q64LH0 and PRF23 were modeled as 3nulA (Q42449) and lg5uB (Q9LE18), respectively. PRF23 exhibited distinct0 three dimensional structural difference in certain fragments compared with Q64LH0 and other Profilins. Though the average values of antigenicity displayed no difference between Q64LH0 and PRF23 on whole se- quences, the antigenicity of PRF23 on certain fragments was obviously lower than that of Q64LHO because of the alteration of some amino acids with different characters, implying the cause of lower incidence of hay fe- ver in South China than in North China. Conclusion Based on sequence analysis, antigenicity evaluation and tertiary structural modeling for Q64LH0 and PRF23, we obtained lots of useful information about the contribution of different amino acids to antigenicity and protein structures, thus would facilitate allergen ge- netic improvement by amino acid replacement.
4.MRS in quantitative assessment of lumbar discs degeneration
Yong LI ; Zhaoxi CAI ; Jianyu CHEN ; Zehong YANG ; Zhenzhen LIU ; Xinhua JIANG ; Mengdi ZHANG
Chinese Journal of Medical Imaging Technology 2017;33(3):440-444
Objective To evaluate the value of MRS in quantitative assessment of degeneration of lumbar discs.Methods Totally 82 patients with lumbago underwent lumbar MR scanning.All the discs were classified with Pfirrmann grades in the sequences of sagittal T2WI.The area under N-acetyl peak,under water peak and the ratio of N-acetyl/Water were measured by MRS.Correlation between MRS values and Pfirrmann grade,age were analyzed.Results In 82 patients,204 lumbar discs were measured by MRS.There were 89,73,39,3 discs in Pfirrmann Ⅱ,Ⅲ,Ⅳ,V respectively.The areas of N-acetyl,water peak and N-acetyl/Water ratio of nucleus region were positively correlated with Pfirrmann grading,respectively (rs =-0.460,-0.204,-0.526,all P<0.05).There were 62,25,37,51,29 discs in patients aged <30,30-39,40-49,50-59,>59 years respectively.The ares of N-acetyl peak,N-acetyl/Water ratio of nucleus region was negatively correlated with the age (rs=-0.247,-0.385,both P<0.05).Conclusion MRS can be used for quantitative assessment of lumbar discs degeneration.
5.Expressions of Fas,FasL,Bcl-2 and Bax proteins in thyroid tissue from patients with Graves′ disease
Jialing WANG ; Yaping ZHAO ; Zhiyong FAN ; Hejun WU ; Wei ZHOU ; Zehong LIU
Chinese Journal of Endocrinology and Metabolism 1985;0(02):-
Objective To investigate the association of the expressions of Fas, FasL, Bcl-2 and Bax in thyroid tissue with the pathogenesis in Graves′ disease (GD). Methods Thyroid tissues from 54 patients with GD and 10 patients with thyroid adenoma (paraadenoma tissue as normal controls) were studied for Fas, FasL, Bcl-2 and Bax expressions in the thyrocytes and lymphocytes by immunohistochemical method. Quantitative analysis was performed by Mias 99 pathological image system. Results (1) The positive granule area, average light density and integrated light density of Fas, FasL, Bcl-2 and Bax in the thyroid tissue from patients with GD were higher than those from normal controls (P
6.Donor ischaemic preconditioning in liver transplantation: A systematic review and meta-analysis
Xingjian ZHANG ; Zhongzhong LIU ; Qi XIAO ; Zehong FANG ; Qifa YE ; Yanfeng WANG ; Yan XIONG
Chinese Journal of Hepatobiliary Surgery 2017;23(3):149-153
Objective To systematically review the effect of donor ischaemic preconditioning in liver transplantation.Methods Databases including the Cochrane Library,PubMed,EMbase,CNKI,VIP and WanFang database were searched up to June 2016 for studies which involved donor ischaemic preconditioning (IPC) in liver transplantation.The data retrieved included 1-year mortality,incidence of Primary Graft Non-Functioning (PGNF),intensive therapy unit (ICU) hospitalization and liver function tests which were used to evaluate the treatment outcomes.The data were analyzed using both the fixed-effect and the random-effects models.For categorical outcomes,risk ratio (RR) with 95% confidence intervals (CI) were calculated.For continuous outcomes,the mean difference (MD) with 95% CI were calculated.The metaanalysis was performed using Review Manager 5.2 software.Results Six clinical studies with 322 patients were qualified for this meta-analysis.There were no significant differences in the 1-year mortality (OR =0.51,95% CI 0.24 ~ 1.05,P > 0.05),PGNF (OR =0.33,95% CI 0.08 ~ 1.40,P > 0.05) and ICU hospitalization (OR =-0.17,95 % CI-2.72 ~ 2.38,P > 0.05) between the donor ischaemic preconditioning and the control groups.There were also no significant differences in the transaminase and bilirubin levels on postoperative day 1,3 and 7 between the two groups.Conclusion There is currently not enough evidence in evidenced based medicine to recommend the routine use of ischaemic preconditioning in donor liver retrieval.
7.Establishment of an anti-CD45 monoclonal antibody that inhibits T-cell proliferation
Zehong SU ; Yalin LI ; Xiangwen TAN ; Xin LIU ; Yueqin YUAN ; Gaojian LIAN
Acta Laboratorium Animalis Scientia Sinica 2014;(5):35-38
Objective To identify molecules that modulate T-cell functions and serve the studies on T-cell media-ted autoimmune diseases.Methods Bone marrow-derived dentritic cells were collected from BALB/c mice to immunize Wistar rats, and to establish many hybridoma cell lines.Many hybridoma cell lines which could modulate T-cell functions were obtained.One of the cell lines, most actively inhibiting T-cell proliferation, was further studied.Results The anti-CD45 mAb recognized CD45 and significantly suppressed T-cell proliferation in proliferation assays.Conclusions Our re-sults indicate that the anti-CD45 mAb can effectively suppress T-cell proliferation, and is promising to be used in the pre-vention and treatment of T-cell mediated autoimmune diseases in the future.
8.Prediction of antigenic epitopes on HA, NA amino acid sequences of novel influenza A (H7N9) virus and analysis association between susceptibility and HLA-Ⅱalleles
Xueting LIU ; Shan WANG ; Junyan ZHANG ; Zhaoyu LIU ; Huifang CHEN ; Zehong ZOU ; Lanyan XIAO ; Zhiheng JI ; Ying HE
Chinese Journal of Immunology 2015;(1):16-21
Objective:To compare the amino acid sequences difference of HA,NA novel influenza virus A/H7N9 isolates, decipher possible B cell epitopes and T cell epitopes of HA,NA protein,and analyze the association between susceptibility and HLA polymorphisms.Methods:The amino acid sequences of novel influenza A ( H7N9) virus were downloaded from Genbank.Phylogenetic trees were constructed based on the amino acid sequences of HA and NA by using software Clustal X and MEGA 4.0.B cell and T cell epitopes were respectively predicted with Protean software and NetMHCⅡ2.2 Server online server.Results:The homology of HA and NA proteins of H7N9 virus was high.10 B cell epitopes and 15 T cell epitopes were randomly distributed throughout HA sequence and 12 B cell epitopes and 9 T cell epitopes were randomly distributed throughout NA sequence.HLA-DRB1*0701 allele which was commonly observed in Northern Chinese population have a high binding affinity for 9-mer peptides of HA and NA proteins.Conclusion:The prediction of B and T cell epitopes of HA and NA proteins with multiple methods benefits the research and development of vaccine against human infection with avian influenza A H7N9 virus.HLA-DRB1*0701 allele may contribute to susceptibility to novel influenza A (H7N9) virus.H7N9 influenza virus is more easily spread in Urumqi,Harbin,Shandong Province,Liaoning Province,Beijing, Shijiazhuang and Tianjin of China.
9.Pathological typing and clinical features in 70 cases of pediatric interstitial pneumonia
Min LI ; Zehong WEI ; Xin YAN ; Tao SONG ; Yuan DONG ; Minzhu ZHAO ; Yunzhi LIU ; Jianbo LI ; Renkuan TANG
Journal of Clinical Pediatrics 2014;(8):727-731
Objective To better understand pathological types of pediatric interstitial pneumonia and improve clinical diagnosis by analyzing the clinical records and pathological typing of interstitial pneumonia. Methods 70 cases of children diagnosed as interstitial pneumonia by autopsy were retrospectively analyzed. Results The number of males was more than that of females. There was a signiifcant predominance of infants less than 2 years. The clinical features include acute onset, rapid development, short duration and atypical clinical manifestations. Most patients had poor prognosis and curative effect with general therapies. Twelve cases had dubious etiology. Pathologic types of 58 cases with unclear etiology were diffuse alveolar damage type (DAD type, 38/58), desquamative interstitial pneumonia type (DIP type, 5/58), lymphoid interstitial pneumonia type (LIP type, 3/58), DAD type complicating DIP type (6/58), DAD type complicating LIP type (2/58), DAD type complicating respiratory bronchiolotitis-associated interstitial lung disease type (RB-ILD type, 3/58), DIP type complicating LIP type (1/58). Conclusions The conifrmed diagnosis rate was relatively low for pediatric interstitial pneumonia. Postmortem examination was helpful for diagnosis and improving clinical diagnosis and pathological typing.
10.Research progresses of lymphopenia induced T cell homeostatic proliferation in antitumor immunity
Nan LAI ; Zehong CHEN ; Zike YANG ; Yanyouhong LIU ; Shijun KANG
Journal of International Oncology 2018;45(2):100-102
Generally speaking,the organism can maintain the stability of T cells.The lymphopeniainduced homeostatic proliferation of T cells could be driven by the recognition of autoantigen including tumor antigen in the absence of foreign antigens or inflammatory signals.This process can break tumor-induced immune tolerance and induce a powerful antitumor immunity.It is confirmed that some negative immune molecules are recruited during the homeostatic proliferation,then the antitumor immunity will be impaired.