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OBJECTIVE@#To explore the laboratory phenotype and molecular pathogenesis in a Chinese pedigree affected with Hereditary coagulation factor Ⅻ (FⅫ) deficiency.@*METHODS@#A male proband admitted to Ningbo No.2 Hospital on July 17, 2021 due to chronic gastritis and members of his pedigree (7 individuals from three generations) were selected as the study subjects. Prothrombin time (PT), activated partial thromboplastin time (APTT), FⅧ activity (FⅧ: C), FⅨ activity (FⅨ: C), FⅪ activity (FⅪ: C), FⅫ activity (FⅫ: C), and FⅫ antigen (FⅫ: Ag) were determined. All of the exons, exon-intronic boundaries, as well as the 5'- and 3'-untranslated regions of the F12 gene were subjected to Sanger sequencing. Candidate variants were verified by cloning sequencing. The effect of candidate variants on the protein function was analyzed by bioinformatics software.@*RESULTS@#The proband, a 47-year-old male, had significantly prolonged APTT (180.0 s) and decreased FⅫ:C and FⅫ:Ag levels (< 1%). His father, mother, brother and two sons also showed certain degrees of reduction. Genetic testing revealed that the proband has harbored compound heterozygous variants of the F12 gene, namely c.1092_1093insC (p.Lys365Glnfs*69) in exon 10 and c.1792_1796delGTCTA (p.Val579Hisfs*32) in exon 14. His mother and elder son were heterozygous for the c.1092_1093ins variant, whilst his father, brother, and younger son were heterozygous for the c.1792_1796delGTCTA variant. Analysis of the promoter region of exon 1 also showed that the proband and both sons had harbored a 46T/T polymorphism, whilst other family members were 46C/T. Bioinformatic analysis suggested that the p.Val579 is a highly conserved site. Protein model analysis showed that, with the p.Val579Hisfs*32 variant, a benzene ring was added and the hydrogen bond of surrounding amino acids was changed. Based on the guidelines from the American College of Medical Genetics and Genomics, the c.1792_1796delGTCTA was classified as a pathogenic variant (PVS1+PM2_Supporting+PM4).@*CONCLUSION@#The c.1092_1093insC (p.Lys365Glnfs*69) and c.1792_1796delGTCTA (p.Val579Hisfs*32) compound heterozygous variants of the F12 gene probably underlay the decreased FXII levels in this pedigree. Above finding has also enriched the mutational spectrum for FⅫ deficiency.
Subject(s)
Male , Humans , Aged , Middle Aged , Pedigree , East Asian People , Exons , Introns , Family , Factor XII Deficiency/genetics , 3' Untranslated Regions , Factor XII/geneticsABSTRACT
BACKGROUND: Maintenance and activation of cascade reaction influence T cell proliferation or transformation into nonreactive state even apoptosis. B7 binding to CD28 effectively activates T cells in combination with T cell receptor pathway, and enhances T cell proliferative activity. OBJECTIVE: To investigate the costimulated activation of peripheral blood mononuclear cells (PBMC) with CD28 and CpG containing oligodeoxynucleotides (CpGODN) MoAb combined with CD80, and its killing effect on human gastric cancerous cell line MKN45 in vitro.METHODS: PBMCs were isolated by Ficoll density gradient centrifugation method, and cocultured with interleukin-2, CD28 and CpGODN MoAb for 1-5 days. MKN45 cells were divided into 4 culture conditions: CD28/CpGODN, CD80 plus CD28/CpG ODN, CD80 alone, and blank control.The killing efficiency was measured by MTT method.The ultramicrostructure of cells was observed by electron microscope. Apoptosis was verified by a flow cytometery. RESULTS AND CONCLUSION: CD80 alone did not display killing effect on MKN45 cells. By MTT method, combination of costimulated activation of PBMC with CD28/CpGODN and CD80 showed enhanced killing effect compared with single therapy (P < 0.05), and the ratio of effector cell and target cell at 15: 1 resulted in half killing efficiency. Electron microscope and flow cytometery verified necrotic or apoptotic cells after 24 hours exposure to costimulated activation. Compared with blank control group, CD80 alone elevated the apoptosis rate of MKN45 cells (P < 0.01). Results from the present study show that CD80 can elevate the killing effect of costimulated activation of PBMC with CD28/CpGODN on MKN45 cells in vitro.
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Objective To explore the value of lymphatic mapping (LM) and sentinel lymph node(SLN) analysis in laparoscopic colectomy for colon carcinoma. Methods Thirty-two patients with clinically localized colonic neoplasms were subjected to submucosal injection of isosulfan blue dye (0.5-1.0 mL) via a colonoscope during operation. Blue-stained lymphatics were visualized through the laparoscope and followed to the SLN,which was tagged. The colectomy was completed in standard fashion. All lymph nodes were stained by hematoxylin and eosin,and multiple sections of each SLN were examined by immunohistochemical (IHC) staining using cytokeratin antibody. Results At least one SLN was identified laparoscopically in all patients. The SLN accurately predicted the tumor status of the nodal basin in 94% of cases. In 8 cases (25%),an unexpected lymphatic drainage pattern altered the extent of mesenteric resection. The SLN was negative by HE staining in 4 (13%) cases,which were demonstrated positive for micrometastases through immunohistochemical staining. Conclusions SLN mapping during laparoscopic colon resection can alter the margins of resection and in combination with immunohistochemical staining may improve staging,which may more accurately assign patients to prospective protocols.
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Objective To investigate an effective method for detecting lymphatic micrometastasis in gastric carcinoma.Methods RT-PCR technique was applied to examine MMP-7 mRNA in 94 lymph nodes in 24 cases of gastric carcinoma.Results Routine pathological method detected lymphatic metastasis in 54 lymph nodes,while MMP-7 mRNA RT-PCR showed positive in 78 lymph nodes. When re-examination for the 28 negative lymph nodes in the initial pathological examination and positive in RT-PCR, 8 lymph nodes with metastasis were found by routine pathological method.2 of 5 patients whose lymph nodes were negitive in pathological examination,but positive in MMP-7mRNA were found liver metastasis 16 and 22 months after radical gastrectomy. Conclusions MMP-7 mRNA RT-PCR is a sensitive method for detecting lymphatic micrometastasis for patients with gastric carcinoma. It is a great help for eluvating the postoperative prognosis and supplemental treatmemt.