Alternative splicing of pre-mRNA is an important mechanism for regulating gene function at the post-transcription level and for producing proteomic diversity in higher eukaryotes. The alternative splicing is regulated by the interaction between diverse cis-acting elements and trans-acting factors. Alternative splicing events of oncogenes, tumor suppressor genes and metastasis suppressor genes are associated with the initiation and development of human neoplasms. The protein isoforms sourced from alternative splicing take part in regulating the gene transcription, cell cycle, apoptosis of cells, and playing a role in tumor growth. It is possible for molecular therapy to target directly isoforms of protein produced by alternative splicing or to interfere with the process of alternative splicing.
Alternative Splicing ; genetics ; Humans ; Neoplasms ; genetics ; RNA Precursors ; metabolism ; RNA, Neoplasm ; analysis ; Transcription, Genetic

Objective To evaluate the safety and efficacy of the valve sparing aortic reimplantation in selected patients with acute type A aortic dissection(AAAD).Methods From October 2012 to March 2014, 65 AAAD patients with entry tear located in the sinus of Valsalva and/or genetic: syndrome underwent emergent operation.Of them, 34 patients had valve sparing aortic reimplantation(David Ⅰ group) , and 31 patients underwent aortic composite replacement(Bentall group).Results No operative mortality was observed in this study.In-hospital mortality(8.8％ vs.9.7％ , P ＞ 0.05) and morbidity (25.4％ vs.27.9％, P ＞0.05) were comparable between two groups.All the patients underwent arch replacement and stented elephant trunk implantation concomitantly.Mean cross-clamp time [(149 ± 23) min v s.(124 ± 21) min, P ＜ 0.05] was longer for David Ⅰ group, while mean cardiopulmonary bypass time[(186 ± 77) min vs.(193 ± 89) min, P ＞0.05] and mean operation time [(341 ± 137) min vs.(378 ± 174) min, P ＞ 0.05] had no significant difference between two groups.The blood transfusion was significantly reduced in David Ⅰ group than that in Bentall group[(1 180 ±490) ml vs.(1 790 ±560) ml, P ＜0.05].The mean follow-up was(17.6 ± 5.4) months(range, 8-26 months).In David Ⅰ group, one patient with genetic syndrome died of ruptured abdominal aortic aneurysm 18 months postoperatively.Two late deaths occurred in Bentall group due to intracranial hemorrhage after 9 months and ruptured infective pseudoaneurysm after 13 months respectively.In David Ⅰ group, average grade of aortic regurgitation 6 months postoperatively was 0.6 ± 0.4.At the latest visit, no pseudoaneurysm on anastomosis was observed.Besides two patients from Bentall group were in NYHA class Ⅱ , all the other patients presented in NYHA class Ⅰ.Conclusion David Ⅰ aortic root reimplantation can be performed safely and obtain excellent short-term results in selected patients with AAAD.Long-term results need continuing follow-up.

Objective:To investigate the clinical, endoscopic and pathological characteristics of synchronous multiple early gastric cancer (SMEGC), and to reduce the rate of missed diagnosis.Methods:Clinical data of 227 early gastric cancer patients treated by endoscopic submucosal dissection (ESD) and/or surgery in Songjiang Hospital, Shanghai Jiaotong University School of Medicine from January 2017 to December 2019 were retrospectively analyzed. The differences of clinical, endoscopic and pathological characteristics between solitary early gastric cancer (SEGC) group (200 cases) and SMEGC group (27 cases) were compared. The relevance of endoscopic and pathological features of major and minor lesions of SMEGC was also analyzed.Results:Among the 227 early gastric cancer patients, 27 (11.9%) were SMEGC (58 lesions), of which 25 cases were detected preoperatively, and 2 cases were reexamined within 6 months after surgery with another lesion found at a different site from the previous lesion. In the SMEGC group, the percentages of male and atrophy and intestinal metaplasia in surrounding mucosa were significantly higher than those of the SEGC group [85.2% (23/27) VS 61.5% (123/200), χ2=5.815, P=0.016; 96.3% (26/27) VS 81.0% (162/200), χ2=3.912, P=0.048]. The mean age of the SMEGC group was significantly higher than that of the SEGC group (68.7±6.7 years VS 63.8±9.8 years, t=-2.561, P=0.011). The correlation analysis showed a significant correlation between the major and minor lesions of SMEGC in the size of lesion ( r=0.640, P<0.001), vertical location ( r=0.518, P=0.006), macroscopic type ( r=0.904, P<0.001) and depth of invasion ( r=0.470, P=0.013). Conclusion:SMEGC is prevalent in elderly males with atrophic gastritis and intestinal metaplasia. It is necessary to be alert to the possibility of multiple cancer lesions, if an early cancer lesion is found under endoscopy, especially those that may have the same or similar shape and invasion depth in the same vertical distribution range.

BACKGROUNDThe role of epigenetics in gene expression regulation and development significantly enhances our understanding of carcinogenesis. All the tumor related genes may be the target of epigenetical or genetic regulation. We selected some epigenetically regulated genes for cDNA array analysis and observed variability in the expression of the DICER1 gene in distinct stages of gastric cancer. The aim of this study was to assess the correlation between the expression of DICER1, an epigenetically regulated gene, and gastric cancer.

METHODSTo detect the expression of 506 tumor-associated genes, including DICER1, in the matched cancerous mucosa, pre-malignant lesion (adjacent mucosa), non-cancerous gastric mucosa and distant lymphocyte metastatic lesion in 3 cases of gastric cancers using cDNA array. DICER1 mRNA expression and DICER1 protein expression were further analyzed by Real-time PCR and Western blot in 32 cases of progressive gastric cancer. DICER1 protein expression was also detected in 33 early and 30 progressive gastric cancers by the immunohistochemistry (IHC) method.

RESULTSIn 3 cases of gastric cancer cDNA array showed dramatically decreased expression of DICER1 in pre-malignant lesion, cancerous mucosa and distant lymphocyte metastatic lesions compared with matched noncancerous gastric mucosa, pre-malignant lesion and cancerous mucosa. Real-time PCR results showed that the expression level of DICER1 mRNA in gastric cancer was significantly down-regulated compared to normal gastric tissue (P < 0.05). The IHC assay also showed that the expression of DICER1 was significantly decreased in progressive gastric cancer. Among the 63 cases of gastric cancers, 13/33 early (39.4%) and 19/30 (63.3%) progressive cancers showed negative expression of DICER1 (50.8%). The difference in expression of DICER1 between early and progressive gastric cancers was significant (P < 0.01). The result of Western blotting showed that DICER1 protein was down-regulated significantly in advanced gastric cancer (P < 0.05).

CONCLUSIONSDICER1 expression is decreased during the progression of gastric cancer, especially in progressive gastric cancers, which indicating DICER1 may play an important role in the development of cancer and the epigenetical regulation involved.

Blotting, Western ; DEAD-box RNA Helicases ; analysis ; genetics ; physiology ; Endoribonucleases ; analysis ; genetics ; physiology ; Epigenesis, Genetic ; Humans ; Immunohistochemistry ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; Ribonuclease III ; Stomach Neoplasms ; chemistry ; etiology ; genetics

OBJECTIVEWith the objective of discovering novel putative chromosomal regions and special genes involved in the carcinogenesis, progression and metastasis of laryngeal squamous cell cancer (LSCC).

METHODSDNA copy profile of LSCC were obtained and analyzed by comparative genomic hybridization (CGH) and a computerized digital image analysis system. cDNA microarray of LSCC was performed and the profile was analyzed by Hierarchical clustering.

RESULTSCGH analysis showed average-12.9 gains and losses of chromosomes in LSCC. Relatively high frequencies of gains were found at 3q15-21 (14/18), 5p12-13 (11/18), 8q22-24 (6/18), 11q12-13 (8/18), 15q21-23 (7/18) and 18p11 (8/18), while those of losses at 1p13-21 (8/18), 3p21-23 (14/18), 5q21-22 (14/18), 9p12-pter (11/18) and 13q21-31 (8/18). Hierarchical clustering analysis showed that the differentially expressed genes were segregated into three groups. Three genes differentially expressed in process I (normal tissue to cancer) and process II (cancer to lymph node metastasis), and the Cy5/Cy3 ratios of twelve genes were either higher than 5.0 or lower than 0.2 in process I or process II. The fifteen special genes were first reported possibly to be the relationships with LSCC. In particular, 4 genes of them, which were cytochrome C oxidase Va, PPBP, EPHX2 and PON1, were first reported to correlate with tumorigenesis. SH3GL2, which was one of the 15 special genes, was located at one of the special chromosome regions, 9p12-pter.

CONCLUSIONThe important genes and special chromosomal aberrances might provide us a clue for further investigation of carcinogenesis, progression and metastasis in LSCC.

Adult ; Aged ; Carcinoma, Squamous Cell ; genetics ; pathology ; Chromosome Aberrations ; DNA, Neoplasm ; analysis ; Disease Progression ; Female ; Gene Expression ; Humans ; Karyotyping ; Laryngeal Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Nucleic Acid Hybridization ; Oligonucleotide Array Sequence Analysis

BACKGROUNDLaryngeal carcinoma is a common malignant tumor of the upper respiratory tract, and in 95% of cases the tumor is laryngeal squamous cell carcinoma (LSCC). The abnormity of SH3-domain GRB2-like 2 (SH3GL2) gene was found in LSCC. In order to clarify the relationship between SH3GL2 gene and LSCC, we evaluated the expression of the SH3GL2 gene in LSCC.

METHODReal-time PCR, immunohistochemistry and Western blotting were used to detect the mRNA and protein expression and find the various rules of SH3GL2 gene in LSCC.

RESULTSThe result of real-time PCR showed that the expression level of SH3GL2 mRNA in LSCC tissue was apparently down-regulated; immunohistochemical analysis showed that SH3GL2 protein was mainly located in cytoplasm, the rate of positive cells and SH3GL2 protein expression level were fluctuated with the pathological classification of LSCC; the result of Western blotting showed that SH3GL2 protein was down-regulated significantly in LSCC samples, especially in metastatic lymph nodes.

CONCLUSIONSThese results suggest that SH3GL2 is a LSCC related gene and its expression level is fluctuated with the pathological classification which indicate that SH3GL2 participates in the development and progression of LSCC. And it may be considered as a novel tumor marker to find both a new anti-oncogene and relative factors of invasion and metastasis of laryngeal carcinoma.

Adaptor Proteins, Signal Transducing ; analysis ; genetics ; Blotting, Western ; Carcinoma, Squamous Cell ; chemistry ; genetics ; Humans ; Immunohistochemistry ; Laryngeal Neoplasms ; chemistry ; genetics ; Polymerase Chain Reaction ; src Homology Domains

OBJECTIVETo explore mechanism of S100A8 in the oncogenesis and development of laryngeal cancer.

METHODSProteins interacting with S100A8 were isolated from laryngeal cancer cell lines Hep-2 by immunoprecipitation assay with anti-S100A8 antibody. The target bands were cut out and identified by maxtrix assisted laser desorption/ionization time of flight (MALDI-TOF). The peptide mass fingerprinting data of the proteins identified were analyzed based on the Mascot database. The NF-kappa B binding sites of the proteins were predicted by P-Match software. The binding ability of one of the proteins to S100A8 was confirmed by co-immunoprecipitation and immunocytochemistry methods.

RESULTSFour proteins interacting with S100A8 were obtained, which were hypothetical protein LOC80154, MHC class I HLA-B, similar to T-box 1 isoform C and sarcolemmal associated protein 1. The four genes were predicted to have NF-kappa B binding sites. MHC class I HLA-B, which is one of targets in NF-kappa B pathway, was first confirmed to have the binding ability to S100A8.

CONCLUSIONThe novel partners of S100A8 identified in the study might be involved in NF-kappa B pathway. The binding ability of MHC class I HLA-B to S100A8 implies that S100A8 might function as a new member with other proteins including HLA-B in NF-kappa B pathway. These findings provide a new clue to further study on the molecular mechanism of S100A8 in the genesis of laryngeal carcinomas.

Animals ; Binding Sites ; Calgranulin A ; genetics ; metabolism ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; HLA-B Antigens ; genetics ; metabolism ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; pathology ; NF-kappa B ; metabolism ; Signal Transduction

OBJECTIVETo investigate the feasibility of multiple displacement amplification (MDA) to apply in the non-invasive prenatal genetic diagnosis of Duchenne muscular dystrophy (DMD).

METHODSMaternal blood was obtained from 20 pregnant women at 7 to 25 weeks of gestation. After the discontinuous density gradient centrifugation with Percoll, the fetal nucleated red blood cells (NRBCs) were stained with Kleihauer test. All positive NRBCs were collected by micromanipulator and then performed with MDA. Sex and short tandern repeat (STR) analysis were determind from a small aliquot of the reaction. The origin of NRBCs was verified and prenatal diagnosis of DMD was made at the same time.

RESULTSThe product length of MDA was >15 kb, while primer extension preamplification (PEP) is only about 1 kb. We completed non-invasive prenatal genetic diagnosis of 6 fetus at high risk of DMD using MDA. The results were all coincident with amniotic fluid control.

CONCLUSIONThe MDA method which provides a highly uniform representation across the genome, representing the entire genome with minimal amplification bias, shows good application prospects.

Erythroblasts ; metabolism ; Feasibility Studies ; Female ; Fetal Diseases ; blood ; diagnosis ; genetics ; Humans ; Muscular Dystrophy, Duchenne ; blood ; diagnosis ; genetics ; Polymerase Chain Reaction ; methods ; Pregnancy ; Prenatal Diagnosis ; methods

Animals ; DNA, Fungal ; Phosphorylation ; RNA Polymerase II ; Sequence Homology, Nucleic Acid ; T-Box Domain Proteins ; genetics

OBJECTIVEFour single nucleotide polymorphisms (SNP) in HOXD10, HOXD12 and HOXD13 genes were chosen to investigate SNP and haplotypes distribution in idiopathic congenital talipes equinovarus nuclear pedigrees.

METHODSGenotypes of 4 SNPs in 84 idiopathic congenital talipes equinovarus nuclear pedigrees were analyzed by restriction fragment length polymorphism and DNA sequencing. Analysis of association between SNP locus and idiopathic congenital talipes equinovarus was performed using ETDT software. Haplotypes and their frequencies in 84 nuclear pedigrees were established and analyzed by TRANSMIT software.

RESULTSrs847151 polymorphism was not detected; the rs847154 located in 5' flanking sequence of HOXD12 gene and the rs13392701 located in exon 1 of HOXD13 gene were noted to have transmission disequilibrium in 84 nuclear pedigrees (P < 0.05).

CONCLUSIONrs847154 located in 5' flanking sequence of HOXD12 gene and rs13392701 located in exon 1 of HOXD13 gene are associated with idiopathic congenital talipes equinovarus; HOXD12 andHOXD13 are important susceptible genes of idiopathic congenital talipes equinovarus.

Adolescent ; Adult ; Child ; Child, Preschool ; Clubfoot ; genetics ; Exons ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Homeodomain Proteins ; genetics ; Humans ; Infant ; Male ; Pedigree ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Young Adult