Study On The Meaning Of The Expression Of Folate Receptor Beta In Tuberculosis Granuloma By Self-Made Tissue Microarray

Background and purposeTuberculosis is the second largest infectious disease in the world, 2014 new TB cases 9.6 million, 150 million deaths, new cases of 120 million HIV positive, 48 million for multi drug resistant tuberculosis, resistance to 9% in multi drug tuberculosis XDR-TB, that is to say, about 5 million TB patients in the present medical condition cannot treat. In low- and middle-income countries, the total burden of TB was $8 billion in 2015.May 19, 2014, who in the World Health Assembly through the "in 2015 the global TB prevention, treatment and control strategies and objectives, the goal is to end global tuberculosis epidemic situation, in 2035 to TB deaths decreased by 95%, occurred to reduce the rate of 90% target, also make the tuberculosis risk families from catastrophic costs.With multi-drug resistant tuberculosis and immune related diseases associated with tuberculosis and immune agents users of tuberculosis, and inhibited by decades of chemical treatment of tuberculosis is the resurgence of the, which gives the broad masses of medical workers for TB treatment and the new challenges; at the same time the world 1 / 3 of the people infected with Mycobacterium tuberculosis, and only 5-10% of infected individuals develop to TB patients and other latent infection in the form of parasitic in the human body, but because of various reasons lead to decreased immunity, may lead to lose control in Mycobacterium tuberculosis, caused by tuberculosis.Pathological tissue microarray is from dozens of first thousands of neatly arranged in on a microscope slide and made into tissue microarray, the technology has been applied in the studies of tumor molecular pathology, the earliest in 1998 by Kononen report, which was mainly used pathological tissue microarray for tumor group of fabric in the DNA, RNA and protein in situ measurement and analysis, after gradually extended to the tissue staining, fluorescence staining, pathological teaching, pathology of standardization training and determination, in situ hybridization and fluorescence in situ hybridization. In particular, the generation of the information processing system of the biological chip, the pathological tissue chip has been able to use the computer automatic analysis. Compared with the traditional pathological slides and pathological tissue microarray can be a lot of fast and automatic processing, which can greatly reduce the manual work, also a large number of pathological tissue under similar conditions were pathological recognition processing, make the diagnosis process more standardized. Pathological tissue chip to improve efficiency, clear diagnosis can also be measured lesion tissue specific gene, mRNA and proteins on the specificity of change, while reducing the need for organizations amount. A large number of results can be obtained in a single experiment.Review of the literature. At present there is no literature reported tuberculosis pathological tissue microarray fabrication and application, this study tries to by collecting a certain number of pathological diagnosis of TB cases in tumor tissue and made tuberculosis pathological chip. And use the pathological chip as a platform for research on TB, relationship of tuberculosis in situ cytokine and Mycobacterium tuberculosis infection, tuberculosis specific genetic changes, and tuberculosis macrophages change, and for tuberculosis diagnosis, treatment and late model in vitro and in vivo experimental models provide a theoretical basis.Methods1. A collection of 2015 hospitalized in the Department of thoracic surgery, affiliated to Fudan University in Shanghai public health center May to December 2016, diagnosis and surgical patients 50 cases as experimental group, including 27 male, 23 female, with a mean age of 33 years, including 18 cases of lymphoid tuberculosis, 12 cases of bone and joint tuberculosis, tuberculosis of vertebral body in 11 cases, 2 cases of chest wall tuberculosis, tuberculosis pleurisy or tuberculosis 7 cases. Preoperative anti-tuberculosis treatment for 0-96 months, excluding the patients with tumor. Strictly following the surgical aseptic principles tuberculosis debridement, in resected specimens in sterile conditions for tissue collection, including tuberculosis necrotic granuloma tissue and adjacent normal tissue. After operation for routine pathological examination, retaining only postoperative pathological acid fast bacilli stain color positive tissue into the group. At the same time, cases of normal tissue(normal tissue 3cm from the edge of the tuberculosis lesion) were collected, and 50 cases were taken as the negative control group. In the positive control group, there were 50 cases of splenic tissue(splenic rupture and other splenic diseases), including 36 males and 14 females, with an average age of 29 years. Saline frozen 50 cases as a blank control group.2. Application of tissue microarray technology to 4 groups respectively according to the order array design into a recipient paraffin block, fusion, slicing, fishing, baking sheets to tissue chip.3. Made of pathological tissue chips were stained with immunohistochemistry, changes in the application of confocal microscope detection of pathological tissue chip Fr beta and other substances, respectively to get the data analysis, observation of tuberculous granuloma, lesions of peripheral tissues and adjacent normal tissues draws the relation between the change of tuberculosis infection state and Fr beta and other substances, experimental data using SPSS statistical software 22.0 analysis, P < 0.05 expression was statistically difference.Results1. Pathology tissue microarray:Appearance: chip points arranged in neat rows, staining distinct, uniform thickness of the tissue section, clear, missing piece rate low, folded tissue rare.Morphological examination: pathological chip because of the smaller slice, there is a certain lack of information theory, but the actual observation and the traditional pathological section is not a big gap.Accuracy: the coincidence rate between the experimental group and the traditional pathological results was 98%, among which there was a case of neck tumor which was not consistent with the traditional pathology.2. Experimental results of immunohistochemistry:Pathological tissue microarray was made on the collected data of patients, and the content of FR- beta and tuberculosis antibody in the tuberculosis foci was significantly increased compared with the negative control group(P<0.05).Conclusion1. The successful production of the first kind of tuberculosis pathological tissue chip, tuberculosis pathological tissue chip production is simple, is expected to become a new type of tuberculosis diagnosis method.2. The upregulation of Fr expression in the tuberculosis foci is related to the infection of Mycobacterium tuberculosis. The molecular targeted therapy of tuberculosis, which takes Fr as the target, is feasible and worthy of further study.3. The concentration of tuberculosis antibody in some cases of tuberculosis is increased, and the change of the antibody in the tuberculosis focus is related to the infection.