ABSTRACT
Objective: To evaluate the value of next generation sequencing (NGS) in the genetic testing of Lynch syndrome. Methods: Immunohistochemical method was used to detect the expressions of DNA mismatch repair (MMR) proteins, including MutL homolog 1 (MLH1), PMS1 homolog 2 (PMS2), MutS homolog 2 (MSH2) and MutS homolog 6 (MSH6) in colorectal cancer, gastric cancer and endometrial cancer tissues collected from Shandong Provincial Hospital between 2016 and 2018. The genomic DNA of 45 patients who were suspected with Lynch syndrome was extracted from non-cancerous tissue paraffin samples, which were postoperatively confirmed by microscope. The mutations of 12 genes including MLH1 and MSH2 were detected using NGS. The germline mutant sites and significance were analyzed by bioinformatics technology and further confirmed by using Sanger sequencing. Results: The immunohistochemical results showed that the 45 cases of suspected Lynch syndrome included 22 cases of MLH1 and PMS2 deficient expression, 16 cases of MLH2 and MSH6 deficient expression, and 7 cases of MMR proteins normal expression. The NGS result showed that 28 cases of adjacent sample from colon cancer patients included 4 cases of MLH1 pathogenic mutation, 1 case of suspected MLH1 mutation, 2 cases of MLH2 pathogenic mutation, 2 cases of suspected MLH2 mutation. No MMR gene mutation was found in adjacent samples of 6 cases of rectal cancer, 6 cases of gastric cancer and 7 cases of colorectal cancer with MMR normal expression. One case of MLH1 or MHL2 pathogenic mutation and one case of MLH1 suspected mutation was detected in adjacent samples of 5 cases of endometrial cancer. Moreover, NGS also detected many other genes mutations and unreported gene mutation sites. Pathogenic and suspected MLH1 and MSH2 mutations were verified by Sanger sequencing. Conclusions: High-throughput NGS is a quick, accurate and reliable technique to identify gene variants in suspected Lynch syndrome patients. It has a wide application prospect for gene testing of tumors associated with Lynch syndrome.
Subject(s)
Colorectal Neoplasms, Hereditary Nonpolyposis , Colorectal Neoplasms, Hereditary Nonpolyposis/diagnosis , Colorectal Neoplasms, Hereditary Nonpolyposis/genetics , DNA Mismatch Repair/genetics , Female , Genetic Testing , Germ-Line Mutation , High-Throughput Nucleotide Sequencing , Humans , Microsatellite Instability , Mismatch Repair Endonuclease PMS2/genetics , MutL Protein Homolog 1/genetics , MutS Homolog 2 Protein/genetics , MutS Homolog 2 Protein/metabolismSubject(s)
Astrocytoma , Brain Neoplasms , Ganglioglioma , Astrocytoma/diagnostic imaging , Astrocytoma/surgery , Humans , Infant , Magnetic Resonance ImagingABSTRACT
Objective: To analyze the expression of SMARCE1 in clear cell meningioma (CCM), and evaluate the role of SMARCE1 in the differential diagnosis in morphologically similar diseases. Methods: Thirteen samples/11 cases of CCMs were collected from the First Affiliated Hospital of Fujian Medical University, Shandong Provincial Hospital, Xuanwu Hospital of Capital Medical University and Thaihe Hospital of Hubei Province from January 2000 to December 2018, as well as 17 cases of meningiomas with clear-cell-like morphology, 782 cases of other types of meningiomas and other intracranial tumors with clear-like morphology. A tissue microarray was made using these cases, on which immunohistochemical/histochemical staining of SMARCE1, SSTR2, EMA, Ki-67, p53, PAS and D-PAS were performed. Result: The tumor cells of CCM had sheet-like architecture, without typical whorl formation.The CCM had round to polygonal cells, with clear, glycogen-rich cytoplasm and prominent blocky perivascular and interstitial collagen. The immunohistochemistry staining showed that none of the CCMs expressed SMARCE1(0/13).However, all of the other types of lesions, including meningioma(782/782), meningiomas with clear-like morphology(17/17), intracranial metastatic clear cell renal cell carcinoma(10/10), haemangioblastoma(10/10), central neurocytoma(10/10), oligodendroglioma(10/10), ependymoma(13/13), lioblastoma(42/42), and solitary fibrous tumor/hemangiopericytoma(35/35) showed positive nuclear staining of SMARCE1. Ki-67 index were 1%-5%, and p53 positive-rate were 0-40% in CCMs. PAS stain showed cytoplasmic granular positive and D-PAS were negative in all CCMs and meningiomas with clear-like morphology. Conclusion: SMARCE1 is a useful marker for the diagnosis of CCM and its mimickers.
Subject(s)
Meningeal Neoplasms , Meningioma , Brain Neoplasms , Chromosomal Proteins, Non-Histone , DNA-Binding Proteins , Diagnosis, Differential , Humans , ImmunohistochemistryABSTRACT
Fumonisin B1 (FB1) is a food-borne mycotoxin produced by genus Fusarium and was classified as possible carcinogen to humans by the International Agency for Research on Cancer (IARC). Human leukocyte antigen (HLA) class I antigen presentation pathway plays an important role in immunosurveillance. Defects in HLA class I antigen presentation pathway can down-regulate the expression of HLA class I antigen on the surface of nucleated cells that will confer a survival advantage to randomly mutant cells and may lead to malignant transformation. In the present study, we analyzed the effects of FB1 on the expression of HLA class I heavy chain (classical HLA-A, -B and -C genes included), beta2-microglobulin (ß2m), LMP2 and TAP1 genes in human gastric epithelial immortalized GES-1 cells in vitro using semi-quantitative Reverse Transcription Polymerase Chain Reaction (RT-PCR), Western blot and immunocytochemical methods in dose- and time-effect studies. Our results revealed that FB1 have an effect on HLA class I antigen presentation pathway via the decreased expression of HLA class I heavy chain and/or defects of LMP2 and TAP1 expression. However, the importance of this effect in carcinogenesis needs further investigation.
Subject(s)
Antigen Presentation/drug effects , Carcinogens, Environmental/toxicity , Fumonisins/toxicity , Gene Expression/drug effects , Histocompatibility Antigens Class I/genetics , Antigen Presentation/genetics , Blotting, Western , Cell Culture Techniques , Cell Line , Cell Survival/drug effects , Dose-Response Relationship, Drug , Epithelial Cells/drug effects , Epithelial Cells/immunology , Humans , Immunohistochemistry , Reverse Transcriptase Polymerase Chain Reaction , Time FactorsABSTRACT
Triboluminescence (TL) is the emission of light induced by the application of mechanical energy to a solid. It links the spectroscopic, structural, mechanical, and electrical properties of solids. Here, materials of ZnS doped with various contents of Mn2+ were investigated. It was shown that they all have favorable triboluminescent property. The effects of the content of Mn2+, the sintering temperature and time on the luminescent property of ZnS:Mn were discussed. It was found the ZnS doped with 1.2% Mn2+ exhibited the strongest TL intensity among the materials investigated. The growth conditions were obtained through experiments, and ZnS:Mn with highly efficient triboluminescence was prepared. The mechanism of triboluminescence is proposed as follows: the electrons are excited from ground state to excited state by mechanic energy, then recombines with holes and give lights. The broad range of exciting energy may contribute to the high triboluminescent efficiency.
Subject(s)
Luminescence , Manganese/chemistry , Silicates/chemistry , Silicon/chemistry , Zinc Compounds/chemistryABSTRACT
This paper presents the recent international progress of the breath diagnosis systems. It discussess the basic principles and method of the breath diagnosis. Especially, the paper introduces the newest progress and the developing trends in diagnosis of the diabetes and gastrointestinal diseases etc. Finally, it introduces the progress of the medical diagnosis expert system and its application in the breath diagnosis based on the electronic nose.
