1.OBSERVATION OF THE KINETIC CHANGE ON IgG,T-CELL SUBSETS,IL-2 LEVEL OF MICE INFECTED TRICHINELLA SPIRALIS
Lijie SHEN ; Shenghua ZHU ; Zhiyong LUO
Chinese Journal of Zoonoses 2000;(6):70-71,66
AimTo explore the kinetic change of the IgG,T-cell subsets and IL-2 level from the mice infected with T. Spiralis. Methods The level of specific IgG A band IL-2 was determined by ELISA,the percentage of CD4+and CD8+ T-cells were examined by flow cytometry on 7, 14,21,28,35 days after mice infected with T. spiralis respectively. ResultAfter infected with T. spiralis,the level of IgG in mice rised gradually,and reached its peak on the 35th day. The change of T-cell subsets showed that CD4+T cells decreased,which CD8+Tcells increased. The ration of CD4+/CD8+cells decreased,and which was the obvious on the 14thday. It did not recover to normal level even on the 35th day. The IL-2 of level reached the peak on the 7th day after infected,then IL-2 level decreased quickly and lower than that of normal mice on 35th day after infected. Conclusion When the acute phase of T. spiralis infection,the immune function of host was inhibited. The protective immunity of against T. spiralis infection was cellular immunity mainly ,in cooperation with humoral immunity.
2.Protective immunity of Trichinella spiralis encapsulated larva antigens
Mingzhi DONG ; Minfang WU ; Lijie SHEN
Chinese Journal of Schistosomiasis Control 2010;22(1):59-62
Objective To compare the immune protective effects of three antigens of Trichinella spiralis encapsulated larvae on mice.Methods The mice were immunized with Trichinella spiralis encapsulated larvae somatic antigen,encapsulated larva excretory-secretory antigen and encapsulated larva surface antigen,3 times with a 7-day interval,and the adjuvant control and normal control group were set up.Seven days after the final immunization,each mouse was orally challenged with 200 Trichinella spirais larvae.The intestinal adult worms and muscle larvae of Trichinella spiralis of each group were recoveried and examined on Day 7 and Day 30 post-challenge,respectively.The level of 8eruln IgG to antigens of Trichinella muscle muscle larvae wa8 detected by ELISA.Results The intestinal adult worms were reduced by 84.89%.89.73%,85.65%.2.57% in the encapsulated larva somatic,excretory-secretory and surface antigen groups,respectively.The muscle lalwae were reduced by 71.71%,80.98%,73.66%,5.60%, respectively.Adtlltwornl reduction rates(P<0.05) and musclelarva reduction rates(P<0.01) of the encapsulated larva excretory-secretory antigen group and surface antigen group were higher than those of encapsulated larva somatic antigen group.The antibody titers in all the immunized groups increased significantly.and the GMRT values of the encapsulated larva somatic,excretory-secretory and surface antigen groups were 32 798.89,3 474.51,2 984.83,respectively,and were 6.09,7.56,6.50 times higher than those of the normal control group(459.32).Conclusions Trichinella spiralis encapsulated larva antigens can induce strong resistance of host to a subsequent challenge infection.Among these antigens,excretory-secretory antigen is more immunogenic.
3.ER stress involves in neuronal degeneration
Lijie FENG ; Haiping WANG ; Yuxian SHEN
Chinese Pharmacological Bulletin 1986;0(04):-
Endoplasmic reticulum(ER) is a locus where proteins are modified,folded and calcium stored.The accumulation of unfolded protein and disorder of calcium ion can lead to ER stress.Early ER stress will induce unfolded protein response(UPR)which may be protective to cells.On the contrary,long-time strong ER stress will induce cell apoptosis,even death.ER stress has been suggested to be involved in some human neurodegenerative diseases.However,the exact contributions of ER stress in the various disease processes are yet unknown.This review mainly summarizes recently reported discoveries concerning ER stress associated with neurodegenerative diseases and highlights current knowledge in this field that may reveal novel insight into disease mechanisms and help to design better therapies for these disorders.
4.Investigate Lymphangiogenesis in Carcinoma of Rectum of Human and Cancer Metastasis
Lijie YAO ; Lei SHEN ; Yang JIANG
Journal of Medical Research 2006;0(08):-
Objective To investigate the effects of vascular endothelial growth factor-C(VEGF-C)and VEGF-R3(fms-like tyrosine kinase-4,Flt-4)on lymphangiogenesis in adenocarcinoma of rectum.Methods A series of 32 adenocarcinoma of rectum were detected for the expression of VEGF-C and VEGF-R3(fms-like tyrosine kinase-4,Flt-4)and Podoplanin by immunohistochemical SABC methods.Results The density of lymphatic vessel in the perimeter of the adenocarcinoma tissue of rectum was much higher than in the interior(P
5.Immune Responses to Challenge Infection in Mice Immunized with Trichinella spiralis Adult Worm Soluble Antigen
Lijie SHEN ; Zhiyong LUO ; Shenghua ZHU ;
Chinese Journal of Parasitology and Parasitic Diseases 1997;0(05):-
Objective To study the immune response induced by the mice immunized with Trichinella spiralis(T.spiralis) adult worm soluble antigen(AWSAg). Methods T.spiralis AWSAg was prepared to immunize Kunming strain mice. Dynamic changes of IgG, IL 2 and T lymphocyte subsets from immunized mice were determined after challenge infection on d 7, d 14 , d 21 , d 28 and d 35 . Results On d 7 after challenge infection, IgG and CD4 + T cells of immunized group were markedly elevated and persisted higher over the observation period. In contrast, CD4 + T cells and CD4/CD8 ratio were significantly decreased in unimmunized group resulted from immune suppression after infection. IL 2 levels reached the peak on d 7 and persisted in high level from d 7-d 21 in both immunized and unimmunized group after infection, then decreased gradually. Till 35 days after infection, IL 2 level was still higher than the normal mice. Conclusion Mice immunized with AWSAg of T. spiralis produced a potential cellular and humoral immune response.
6.Effect ofα1-antitrypsin Z variant overexpression on autophagy regulation
Na ZHU ; Lijie FENG ; Haiping WANG ; Yujun SHEN ; Yuxian SHEN
Chinese Pharmacological Bulletin 2014;(7):921-925
Aim To investigate the effect of α1-anti-trypsin Z variant (ATZ)overexpression on cell autoph-agy.Methods HEK 293T cells were transfected with pcDNA3.1 zeo+/ATM or pcDNA3.1 zeo+/ATZ,e-qual amount of empty vector was used as control.Cells were treated with NH4Cl for 4 hours and processed for detecting ATZ,LC3 and p62 by immunoblot.Mean-while ,expression and intracellular localization of ATZ, LC3 in 293 T cells were observed with double labeled immunofluorescence.The mRNA levels of autophagy-related genes were measured by real-time PCR.Immu-nohistochemistry was used to observe the morphology of ATZ-positive cells.Results Compared with the control,higher LC3Ⅱ levels and LC3 puncta were observed in ATZ transfected cells.Meanwhile,the levelsof p62 were decreased in ATZ transfected cells,andreversed by NH4 Cl (25 mmol·L -1 )treatment.Overexpression of ATZ increased the mRNA levels of Atg5and Atg12,but had no obvious influence on Beclin1.ATZoverexpressing cells presented abnormal morphologies.The nuclei became reduced,condensed,and even disappeared in ATZpositive cells.Conclusion ATZ overexpression increases autophagy activity whichmay be related to increasing Atg5 and Atg12 levels.
7.Expression of RNF2 in breast carcinoma and its significance
Qing LI ; Zhengsheng WU ; Yujun SHEN ; Lijie FENG ; Yuxian SHEN
Chinese Journal of Clinical and Experimental Pathology 2017;33(3):237-240
Purpose To investigate the expression of RNF2 in breast disease tissues and cell lines,and to analyze the association between expression of RNF2 and clinicopathological characteristics in breast carcinoma.Methods Expression of RNF2 protein and mRNA levels was detected using immunohistochemistry of EnVision two-step and qRT-PCR in breast carcinoma and benign breast disease as well as in cell lines.Results RNF2 expression was sigmificantly higher in breast carcinoma tissue specimens compared with benign breast disease specimens (P <0.05).Besides,the expression of RNF2 protein was significantly associated to tumor size,lymph node status and TNM stage (P < 0.05 for both),but was not related to age,histological grade,the expression of ER,PR and HER-2 (P > 0.05 for both).Higher expression of RNF2 mRNA was detected in breast carcinoma cell lines compared with breast epithelial cell lines (P < 0.05).Conclusion RNF2 is overexpressed in breast carcinoma and can be a potential therapeutic target for breast carcinoma.
8.Study and application of surface antigen in tachyzoites of Toxoplasma gondii
Li LENG ; Mi LUO ; Ju GAO ; Lijie SHEN
Chinese Journal of Schistosomiasis Control 2014;(6):687-689
Toxoplasma gondii is an intracellular protozoan parasite that infects all warm?blooded animals. The surface anti?gens of T. gondii with the potential for application as antigens of diagnosis and vaccines have been studied extensively in recent years especially for P43 P35 P30 P23 and P22. The studies on the surface antigen in tachyzoites of T. gondii are reviewed in this paper.
9.Autophagy alleviates neuronal toxicityinduced by abnormally phosphorylated tau protein
Qian DING ; Jin ZHANG ; Yuyang MA ; Yujun SHEN ; Yuxian SHEN ; Lijie FENG
Chinese Pharmacological Bulletin 2017;33(6):761-767
Aim To observe the effects of rapamycin (Rapa) and starvation-induced autophagy on the morphology of neuronal cells, tau protein aggregation and expression of phosphorylated tau protein, to explore the possible mechanism of cytoprotective effect of these two classical autophagy inducers on phosphorylated tau expressing cells.Methods N2a cells were transfected with GFP-tau plasmid, and equal amount of empty vector was used as control.Then cells were incubated with or without okadaic acid(OA) for 12 h, followed by treatment with autophagy inducers rapamycin(Rapa) and EBSS, autophagy inhibitor Bafilomycin A1(Baf A1) for 6 h.DAB was used to observe tau expression and cell morphology.Confocal microscopy was used to observe the intracellular tau aggregation.TUNEL assay and cleaved caspase-3 level were used to detect cell apoptosis.Immunoblot was used to detect the expression of phosphorylated tau and autophagy-related proteins.Results Our study showed that the N2a cells treated with OA exhibited small cell body, retracted processes and increased tau aggregation, compared with only tau-expressing cells.Rapa and EBSS treatment significantly improved cell morphology, decreased tau aggregation and reduced cell apoptosis.On the contrary, Baf A1 treatment induced aberrant cell shape and increased tau aggregation and cell apoptosis.In addition, Rapa significantly decreased the high molecular weight, phosphorylated tau whereas EBSS especially decreased the low molecular weight phosphorylated tau.Conclusions Rapa and EBSS is alleviate hyperphosphorylated tau-induced cytotoxicity through different mechanism.Rapamycin mainly decreases phosphorylated tau oligomers, while EBSS liable to decrease the soluble phosphorylated tau.
10.Expression and methylation status of CAV-1 gene in esophageal squamous cell carcinoma
Zhen ZHOU ; Yanli GUO ; Lijie HAN ; Wei GUO ; Shumei LI ; Supeng SHEN ; Zhiming DONG
China Oncology 2014;(10):789-793
Background and purpose: As one of the important epigenetic phenomena, DNA methylation plays an important regulatory function for the expression of genes. Study shows that abnormal changes of DNA methy-lation patterns of normal tumor cell genome leads to dysfunction of cancer related gene, and this may be associated with tumor occurrence and development. The study investigated the promoter methylation and expression of caveolin-1 (CAV-1) gene in esophageal squamous cell carcinoma (ESCC), and to elucidate its role in ESCC. Methods:We used MSP approach, RT-PCR, and immunohistochemistry method respectively to examine the methylation status of the 5’CpG island of CAV-1 gene and its expression at mRNA and protein levels in tumors and corresponding normal tissues. Results: CAV-1 mRNA expression in tumor tissues (0.86±0.56) was signiifcantly higher than that in corresponding normal tissues (0.40±0.36, P<0.05). The mRNA expression of CAV-1 was correlated with status of lymphatic metastasis and TNM stage of ESCC patients (P<0.05). The protein expression of CAV-1 in tumor specimens (66.7%, 34/51) was signiifcantly higher than that in corresponding normal tissues (15.7%, 8/51, P<0.01). The protein expression of CAV-1 was signiifcantly associated with lymphatic metastasis of ESCC (P<0.05), however, it was not associated with differen-tiation and TNM stage (P>0.05). The promoter methylation frequency of CAV-1 in tumor specimens was 2.0%(1/51), and the methylation phenomenon has not been found in corresponding normal tissues. The promoter methylation fre-quency of CAV-1 in tumor specimens showed no signiifcant difference compared with the corresponding normal tissues (P>0.05). Conclusion:The mRNA and protein expression of CAV-1 in tumor specimens was signiifcantly higher than that in corresponding normal tissues. Aberrant high expression of CAV-1 has played a certain role in promoting tumori-genesis and lymph node metastasis. The expression both in ESCC and corresponding normal tissues has no correlation with the promoter methylation status.