| ObjectiveDiscussion matrix protein Fibulin-5 fibrosis in normal lacrimal gland and orbital IgG4-related diseases of inflammation and hardening.MethodsCollected from March 2014 to June 2016 with orbital IgG4-related diseases of Orbital lacrimal gland tumor removal surgery in the People’s Liberation Army 474 hospital.,a tolal of 20 cases with 20 eyes(IgG4-ROD group)from acquisition of orbital lacrimal gland tumor resection.While collected from 20 cases with 20 cases of normal lacrimal gland tissue specimens(open-ended comminuted fracture surgery)as control group.All patients had no immunization and chemotherapy before surgery and were comparable in age(P> 0.05).After obtained the consent of the hospital ethics committee and were obtained the patient’s informed consent,all the organizations in the surgery were removed from the orbital lacrimal gland tumor.The lacrimal gland tissue specimens are divided into three sizes are about 0.3cm × 0.3cm × 0.3cm with saline fully washed.Take a piece of tissue into the 4% paraformaldehyde fixed 24 h,and then removed into the automatic tissue dehydration machine,followed by paraffin embedded,made of wax block back,the other two into the tin foil wrapped and equipped with Trizol EP tube,put Into-80 ℃refrigerator to save spare.1.The expressions of Fibulin-5 and mRAN were detected by real-time fluorescence quantitative PCR and the expression of Fibulin-5 and protein were detected by Western blot.The data were analyzed by SPSS19.0 statistical software.Shapiro-Wilk test was normal distribution,the results of t test,mean ± standard deviation(X ± s)that the number of groups by Levenne test to P <0.05 was significantly different.2.The expression of Fibulin-5 and protein was detected by immunohistochemical method.The expression of Fibulin-5 and protein was evaluated by Fisher’s exact rate method.The correlation test was performed by Sperman correlation.Sex analysis,with P <0.05 was significantly different.Results1.Real-time fluorescence quantitative PCR detection: Orbital IgG4-related disease compared with patients with normal lacrimal gland,orbital IgG4-related disease lacrimal gland tissue Fibulin-5mRNA significantly higher than normal lacrimal glands.The expression level of Fibulin-5 mRNA in IgG4-ROD group was 3.69 ± 1.72 and 1.69 ± 0.45 in the control group.The difference was statistically significant(P <0.05)according to the test level of P <0.05.2.Western blotting showed that the expression of Fibulin-5 protein and Fibulin-5 mRNA were consistent.Orbital IgG4-related disease compared with normal lacrimal gland patients,orbital IgG4-related disease lacrimal gland tissue Fibulin-5 protein significantly higher than normal lacrimal glands.The expression of Fibulin-5 protein in the IgG4-ROD group was 0.55 ± 0.12,while that in the control group was 0.19 ± 0.11,and the difference was statistically significant(P <0.01)).3.Immunohistochemical staining showed that Fibulin-5 protein was mainly distributed in the cytoplasm of the cells,and the positive staining was yellow or brown.The results of immunohistochemistry showed that the lacrimal gland cells in the IgG4-ROD group were irregularly arranged and stained with yellow and brown fibrous cells.In the control group,the laryngeal ducts were arranged in the control group,and the cells were arranged in blue.The positive rate of Fibulin-5 protein in the IgG4-ROD group was 80%(16/20).The positive rate of Fibulin-5 protein in the control group was 22.5%(9/40).According to the test level of P <0.05,the difference between the two groups was statistically significant The positive expression of Fibulin-5 protein in IgG4-ROD group was significantly higher than that in control group.Conclusion1.The expression of Fibulin-5 mRNA and protein in the lacrimal gland of the orbital IgG4-related disease was significantly higher than that in the normal lacrimal gland compared with the normal lacrimal gland in patients with Fibulin-5 mRNA and protein levels. |
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