Immunological surveillance, analysis including cells to biological is one of the important components in clinical laboratory immunology. This paper focuses on three issues. First of all is the measurement of antigen-specific T cell, which is very important for understanding the pathogenesis of immunological related diseases because of the central role played by T cell in immune response. It is optional to adopt different assays according to different facilities and puroposes such as flow cytometer or the number or function of T cells. Secondly, it is no doubt that positive autoantibodies are key bio-markera in the diagnosis of autoimmune diseases. And what is more autoantibodies could predict the occurrence of certain autoimmune diseases. More and more researches showed that autoantibodies could be detected in patients with tumors, which help for either diagnosis of tumors or screening of therapeutic target markers. Finally, the application of cytokine measurement is discussed. Clarification of cytokine profiles is the way to understanding pathogenesis of immunological related diseases, it is also in favor of monitoring severity of the diseases, But there are shortages concerning organ or tissue specific, as well as good quality control.

MicroRNAs (miRNAs) are a new class of noncoding RNA which regulate gene expression at post-transcription level.It is believed that miRNAs are widely involved in the pathological and physiological processes,including the proliferation,development,differentiation and apoptosis regulation.

Autoantibodies are useful laboratory parameters for diagnosis of autoimmune diseases (AID).Methods for the detection of autoantibodies have achieved quantitative or semi-quantitative to some extent.Furthermore,with the development of detection technology,high-throughout and quantitative technology has been the trend in autoantibody measurement.Compared with qualitative results,the quantitative ones may provide more values for diagnosis,prediction,prognosis and therapeutic monitoring for AID.Therefore,although quantitative technology of autoantibodies is still faced a number of challenges,the detection for autoantibodies has come into the era of quantitation.

Primary biliary cirrhosis(PBC) is an autoimmune disease with unknown causes.Most PBC patients have abnormal lipid metabolism characterized by hypercholesterolemia.Paradoxically,clinical observations and pre-experimental studies showed that the risk of hyperlipidemia associated cardiovascular event and the mortality of PBC patients were not increased.In this review we summarize the possible reasons and the underlying mechanisms.

Autoimmune diseases (AID) are a group of diseases,in which the tolerance of immune system to self component is broken.However,the etiology and pathogenesis of AID has not yet been clear so far.For better understanding the pathogenesis of AIDs and providing new idea on the diagnosis and treatment of AID,this review will focus on the latest development on pathogenesis of AID,including genetic background,environment factors,abnormal immune regulation,and the role of target cells.

Laboratory diagnostics is an important discipline functioning as a bridge between basic and clinical medicine and it is closely associated with the diagnosis of clinical physician.But there are some problems in the laboratory diagnostic teaching including unreasonable curriculum standard,simple teaching method and unpractical theory.This paper explored and summarized the problems and the reform of laboratory diagnostics teaching mode for students of clinical medicine.

Objective:In view of analysis of role of autoantigen Dsg3 in specific T cell response to understand the molecular mechanism of autoimmune diseases.Methods:Based on the sequence analysis of autoantigen Dsg3,the five fractions cDNAs of Dsg3 were cloned and Dsg3-GST fusion proteins induced by IPTG in E.coli.JM109 were purified,then mixed-cultured with T cell from PV patients.The T cell proliferation response were analyzed by MTT.Results:Dsg3 E1,E2 and E4,E5 stimulated T cell from PV patients,not controls.Conclusion:Dsg3 E1,E2 and E4,E5 contained the epitopes relevant to T-B cell interaction,which play an important role in the pathogenesis of pemphigus vulgaris.

The present study was to analyze the epitopes of autoantigen Dsg3 in pemphigus vulgaris (PV), and to elucidate the molecular mechanisms underlying autoimmune diseases. Based on the sequence analysis of autoantigen Dsg3 in Genbank five fragments of Dsg 3 cDNA (E1, E2, E3, E4, and E5) were cloned using RT PCR. Then PCR products were inserted into the expression vector pGEX 2T, and transformed into JM109 strain of E. coli. Dsg3 recombinant proteins were purified from the cell lysates. The epitopes of Dsg3 in PV patients were analyzed with five Dsg3 recombinant proteins by Western blot. The results showed that Dsg3 E1, E2 and E4 recombinant proteins reacted with PV patients′ sera, but not with control or normal sera. The dominant epitopes of Dsg3 in PV patients were different. The above data indicated that the epitopes of Dsg3 were located in E1, E2, and E4, which might play an important role in the pathogenesis of pemphigus vulgaris.

Objective To express the immunodominant epitopes of the branched chain 2 oxo acid dehydrogenase complex (BCOADC), the pyruvate dehydrogenase complex (PDC), the 2 oxo glutarate dehydrogenase complex (OGDC) and a triple hybrid clone (designated as BPO), and use BPO as a tool for the detection of M2 specific for primary biliary cirrhosis (PBC). Methods The cDNA fragments encoding the M2 reactive epitopes of BCOADC, PDC and OGDC were amplified using PCR with total RNA extracted from human peripheral mononuclear blood cell. The fragments were cloned into pQE 30 and then transformed into plasmid E.coli M15. Its products were induced by isopropylthio ? D galactoside and confirmed with SDS PAGE and Western blot. Results Four specific proteins induced from the transformants containing the fused plasmid were shown to have antigenic reactivity with PBC sera, but not with normal control sera. Conclusions We succeeded in expressing three immunodominant epitopes and a hybrid clone which can be used as a powerful and specific method for the diagnosis of PBC.

Objective By in vitro culture of mouse macrophage cell line RAW264. 7 and primary mouse bone marrow macrophages, the expression of Siglec-1 when stimulated by ox-LDL was observed. Meanwhile, Siglec-1 was up-regulated by M-CSF and down-regulated by small interference RNA targeting Siglec-1 ( si-RNA-Siglec-1) , and the expression of chemokines and lipid uptake ability by macrophages were observed, to explore the role of Siglec-1 on macrophages in atherosclerosis. Methods LDL was oxidized by copper. According to preliminary experiment results, ox-LDL 100 μg/ml was selected as a stimulus. There were 6 experimental groups:normal control group,ox-LDL 100 μg/ml group, ox-LDL 100 μg/ml + si-RNA 2509 2 ng/ml group,ox-LDL 100 μg/ml + si-RNA 3618 2 ng/ml group,ox-LDL 100 μg/ml + M-CSF 5 ng/ml group and ox-LDL 100 μg/ml + M-CSF 10 ng/ml group. si-RNA-Siglec-1 was transfected into macrophage to inhibit the expression of Siglec-1, whereas M-CSF 10 ng/ml or 5 ng/ml were added into the culture medium to enhance the expression of Siglec-1. Quantitative real-time polymerase chain reaction ( qRT-PCR) was used to determine the interfere efficiency of si-RNA-Siglec-1 or M-CSF. After stimulation with ox-LDL for 48 h, cell culture supernatants were collected to determine MIP-1 alpha, MCP-1 and IL-8 concentration by ELISA (n =3 for each group) to evaluate the activation of macrophages. Internalization of lipid particles by macrophages was analyzed by oil red 0 staining. Results Observed by fluorescence microscope, si-RNA-Siglec-1 could be effectively transfected into macrophages with a transfection efficiency about 90% ;PCR results showed that si-RNA 2509 and si-RNA 3618 in a concentration of 40 pmol/L had an inhibition rate of 0. 54 ±0. 11 or 0. 52 ±0. 16 vs 1. 00 ±0. 24 (control group) , t =5. 227 and 4. 992, respectively, all P < 0.01, while M-CSF 10 ng/ml could increase Siglec-1 mRNA expression approximately 4-fold (4. 16 ± 1. 25 vs 1.00 ±0. 24, t =7. 448, P<0. 01). The secretion of MCP-1, MIP-1 alpha, and MIP-2 in si-RNA3618-Siglec-1 group [(359. 28±47. 80) pg/ml, (33. 76 ± 14. 28) ng/ml and (7.87±1.55) ng/ml for MCP-1,MIP-1 alpha, and MIP-2, respectively] was significantly reduced in compare with ox-LDL 100 μg/ml group [ (577. 89 ± 35. 95 ) pg/ml, (69. 17 ± 11. 82) ng/ml and (12.28 ± 1.19) ng/ml for MCP-1, MIP-1 alpha, and MIP-2, respectively], with P value of 0.01, 0.05 and 0.01. In contrary, ox-LDL 100 μg/ml plus M-CSF 10 ng/ml group could significantly promote macrophage chemokine secretion [ (672. 89 ± 43.80) pg/ml, (101.31 ±24.17) ng/ml and (14.81 ±0.54) ng/ml for MCP-1, MIP-1 alpha, and MIP-2, respectively], with P < 0.05 compared with ox-LDL 100 μg/ml group. Meanwhile, lipid intemalization and foam cell formation was inhibited in si-RNA3618-Siglec-l group while ox-LDL 100 μg/ml plus M-CSF 10 ng/ml group could enhance the phagocytosis of ox-LDL by macrophage. Conclusions Siglec-1 may served as a potential phagocytic receptor for ox-LDL involving in macrophage uptake of lipid and turn into foam cells. Furthermore, it can active macrophages and enhance the secretion of MIP-1 alpha, MCP-1 and IL-8, attracting more macrophages and lymphocytes to the site of inflammatory plaque. Targeted inhibition of Siglec-1 reduces macrophage uptake of lipid and secretion of chemokines. Siglec-1 may possibly serve as a potential target of treatment or delay the development of atherosclerosis.