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Objective@#To detect the expression and the differential significance of CK5/6, DSG3, P40, TTF-1, CK7, NapsinA in small biopsy specimens of non-small cell lung cancer (NSCLC), squamous cell carcinoma (SCC) and adenocarcinoma (AC).@*Methods@#Immunohistochemical SP method was used to detect the expressions of CK5/6, DSG3, P40, TTF-1, CK7 and NapsinA in 120 small biopsy specimens of NSCLC hospitalized in the Central People's Hospital of Tengzhou from January 2016 to December 2017, and the results were analyzed combined with the clinical characteristics of NSCLC.@*Results@#The positive expression rates of CK5/6, DSG3 and P40 in lung SCC were 100.0%(56/56), 89.3%(50/56) and 96.4%(54/56), respectively, with specificity of 90.6%, 100.0% and 100.0%, respectively.The positive expression rates of NapsinA, TTF-1 and CK7 in lung AC were 81.3%(52/64), 90.6%(58/64) and 93.8%(60/64), respectively, with specificity of 100.0%, 92.9% and 96.4%, respectively.The positive expression rates of CK5/6, DSG3, P40 in SCC had statistically significant differences compared with those in AC (all P<0.05), and the expression of TTF-1, CK7 and NapsinA in AC had statistically significant differences compared with those in SCC (all P<0.05).@*Conclusion@#CK5/6, DSG3, P40, TTF-1, CK7 and NapsinA are of great significance in the differential diagnosis of SCC and AC in small biopsy specimens of NSCLC.
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Objective To detect the expression and the differential significance of CK 5／6,DSG3,P40,TTF-1,CK7,NapsinA in small biopsy specimens of non -small cell lung cancer ( NSCLC),squamous cell carcinoma (SCC) and adenocarcinoma (AC).Methods Immunohistochemical SP method was used to detect the expressions of CK5／6,DSG3,P40,TTF-1,CK7 and NapsinA in 120 small biopsy specimens of NSCLC hospitalized in the Central People's Hospital of Tengzhou from January 2016 to December 2017,and the results were analyzed combined with the clinical characteristics of NSCLC.Results The positive expression rates of CK5／6,DSG3 and P40 in lung SCC were 100.0%(56／56),89.3%(50／56) and 96.4%(54／56), respectively,with specificity of 90.6%,100.0% and 100.0%,respectively.The positive expression rates of NapsinA ,TTF-1 and CK7 in lung AC were 81.3%(52／64), 90.6%(58／64) and 93.8%(60／64),respectively,with specificity of 100.0%,92.9% and 96.4%,respectively. The positive expression rates of CK5／6,DSG3,P40 in SCC had statistically significant differences compared with those in AC (all P＜0.05),and the expression of TTF -1,CK7 and NapsinA in AC had statistically significant differences compared with those in SCC ( all P＜0.05).Conclusion CK5／6,DSG3,P40,TTF-1,CK7 and NapsinA are of great significance in the differential diagnosis of SCC and AC in small biopsy specimens of NSCLC .
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Objective To investigate The Th1/Th2 and Tc1/Tc2 polarization in the peripheral blood of first degree relatives of pemphigus vulgaris(PV) and healthy control individuals, and to approach the mechanism of the Dsg3-specific autoimmunity in PV. Methods The peripheral blood mononuclear cells (PBMC) from first degree relatives and healthy control was stimulated for72 h with Dsg3 and without Dsg3. Th1/Th2, Tc1/Tc2 was assessed by four-color flow cytometry. Results The mean frequency of Dsg3-spe-cific Th2 cells for PV antibody positive first degree relatives was 10.13%±3.72%, compared with stimula-tion without antigen 7.28%±3.58%, the difference was significant (P<0.05). The percentage Dsg3-spe-cific Th2 was markedly higher in the PV antibody positive first degree relatives group than that in the control group(10.13%±3.72% vs 6.10%±2.82%, P<0.05) , Tc2 was markedly higher also (20.01%± 10.43% v514.91%±8.06%, 20.01%±10.43% vs 9.58%±5.49%, P<0.05). Conclusion When Dsg3 stimulated PBMC were used to stimulate autologous T cells an increased amount of Th2 and Tc2 was observed, it is implied that the imbalance of Th1/Th2, Tc1/Tc2 might play an important role in the initia-tion of PV.
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Objective To study the effect of single chain variable fragment (ScFv) antibody to EC3-4 fragment of desmoglein (Dsg) 3 in a mouse model of pemphigus vulgaris.Methods The ScFv an- tibody to EC3-4 fragment of Dsg-3 was injected subcutaneously into neonatal BALB/c mice at different time points;the mice were then evaluated clinically,histopathologically and by direct immunoflorescence exami- nation for the development of lesions.Results When injected alone,the ScFv antibody did not induce the appearance of key clinical features of pemphigus vulgaris.The antibody also did not prevent the develop- ment of pemphigus vulgaris features induced by sera of patients with pemphigus vulgaris,regardless of the time point of injection of ScFv antibody.Conclusion The ScFv antibody to EC3-4 fragment of Dsg-3 lacks pathogenicity in neonatal BALB/c mice,and also could not inhibit the development of lesions induced by sera from patients with pemphigus vulgaris.
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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.
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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.