Clinical analysis for the application of endoscopic ultrasonography in the diagnosis of patients with a retroperitoneal space-occupying lesion

To report our experience and to evaluate the application of endoscopic ultrasonography [EUS] in the qualitative diagnosis of retroperitoneal space-occupying lesions. Twenty-six patients with retroperitoneal space-occupying lesions confirmed by CT or MRI were studied. All the patients underwent endoscopic ultrasonography guided fine needle aspiration [EUS-FNA] at the Department of Gastroenterology, Xiangyang Central Hospital, Hubei Province, China from August 2009 to August 2011. Different parameters were evaluated, such as complications of EUS-FNA, the ratio of definite pathological diagnosis, and the pathologic types of all the specimens. Of the 26 patients, 24 had definite pathological diagnosis; the ratio of histodiagnosis was 92.3%. There were no complications such as hemorrhage, infection, or injury to the abdominal viscera in the process of EUS-FNA. Eight patients had benign tumors, with a ratio of 33.3%, and 16 patients had malignant tumors, with a ratio of 66.7%. Two patients had no definite pathological diagnosis because of the shortness of tissue sample. Eight patients did not undergo operation due to the diagnosis of benign tumors. The EUS-FNA has the advantage of lower complications, and higher diagnostic ratio, which is valuable in the qualitative diagnosis of retroperitoneal space-occupying lesions

relationship among human papilloma virus infection, survivin, and p53 gene in lung squamous carcinoma tissue

To study the relationship between the infection of human papillomavirus [HPV] type 16, type 18, the expression of surviving, and the mutation of p53 gene in lung squamous carcinoma tissue for the research of pathogenesis of lung carcinoma. This study was carried out at the Laboratory of Molecular Biology, Xiangfan Central Hospital of Hubei Province, China from September 2008 to May 2010. Forty-five specimens of lung squamous carcinoma tissue confirmed by histopathology were the excisional specimens taken by the Thoracic Surgery of Xiangfan Central Hospital. Normal tissue, closely adjacent to the fresh carcinoma specimens, was used as the control group for p53 gene mutation analysis. Sixteen surgical excisional specimens of benign lung disease were used as a control group of non-carcinomatous diseases. human papillomavirus DNA were detected by polymerase chain reaction [PCR], and we used the PCR-single-strand conformation polymorphism-ethidium bromide [PCR-SSCP-EB] method to detect the mutations of the p53 gene. The expression of the surviving gene was detected by immunohistochemistry methods. Approximately 68.9% of 45 lung squamous carcinoma tissue had p53 gene mutations. The mutation rate of exon 5-8 p53 were 15.6%, 17.8%, 15.6% and 20%. Approximately 42.2% of lung squamous cell carcinoma samples were shown to be positive for HPV DNA expression and 62.2% were positive for surviving expression. There was an inverse correlation between the presence of HPV infections and mutaions of p53 gene; and the mutations of p53 gene and expression of surviving had a positive relationship. Mutation of p53 gene and HPV infection may facilitate each other in the generation of lung squamous cell carcinoma. abnormal expression of the surviving gene may take part in the onset and progression of lung squamous cell carcinoma

Construction and expression of anti-CD62P/anti-GPIIb-Illa diabody

To construct 3 expression plasmids for the targeted therapy of thrombosis: single chain variable fragment [scFv] of monoclonal antibody [mAb] 7E3 that can identify and bind platelet glycoprotein GPIIb-IIIa complex, scFv of mAb WAPS12.2 that can identify and bind P selectin [CD62P], a diabody that can identify and bind GPIIb-IIIa and CD62P imultaneously, and to investigate whether the vectors can express correctly. This study was carried out at the Laboratory of Hunan Yuantai Biological Technology Co. Ltd, Hubei, China from September 2007 to May 2008. Total RNA of mAb 7E3 cells and WAPS12.2 cells were obtained. Reverse transcriptase polymerase chain reaction [PCR] was carried out to obtain the genes of variable regions of light and heavy chains of 7E3 and WAPS12.2. Target genes were named 7E3VL, 7E3VH, CD62PVL, and CD62PVH. The 7E3VL-7E3VH and CD62PVL-CD62PVH were obtained by PCR and connected with pET-22b[+]. Products were named pET-scFv7E3 and pET-scFvCD62P. The 7E3VL-CD62PVH and CD62PVL-7E3VH were obtained by using overlap PCR and were ligated to pET-22b[+]. The products were named pET-ED1 and pET-ED2. The PCR was performed by taking pET-ED2 as a template to obtain the complete operon gene and was ligated to pET-ED1. The product was named pET-7ECD. The identification by restriction endonuclease cleavage and DNA sequencing confirmed that the construction of these expression plasmids was successful. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot confirmed that these plasmids expressed correctly. The expression plasmids pET-scFv7E3, pET-scFvCD62P, and pET-7ECD were constructed and expressed successfully, and laid a good foundation for further research on target-oriented thrombolytic agents

Construction of adeno-associated virus coexpression system for human angiopoietin-1 and VEGF gene / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Ischemic disease is one of the leading causes of death in the world. In order to further study gene therapy for ischemic disease, we constructed a recombinant plasmid for co-expression of human angiopoietin-1 and vascular endothelial growth factor 165(VEGF165) gene in adeno-associated virus (AAV) gene delivery system.</p><p><b>METHODS</b>Human angiopoietin 1 and VEGF165 gene were obtained using PCR. The upstream of angiopoietin 1 contained restriction enzyme site HindIII, and the downstream of angiopoietin 1 contained restriction enzyme site BamHI. The upstream of VEGF165 contained restriction enzyme site BglII, and the downstream of VEGF165 contained restriction enzyme site BamHI. Using the multiple cloning sites (MCS) in plasmid pZero++ such as BamHI, BglII, HindIII, NotI, XhoI, XbaI, SalI, BspHI, KspI and the corresponding MCS in plasmid pAAV-MCS, angiopoietin 1 and VEGF165 gene were subcloned into pAAV-MCS.</p><p><b>RESULTS</b>DNA sequencing revealed that the PCR- amplified angiopoietin 1 and VEGF165 were consistent with NCBI Gene Bank. The recombinant plasmid was identified using PCR and digestion, which proved to be consistent with our hypothesis. In recombinant plasmid, angiopoietin1 and VEGF possessed a CMV promoter and polyA terminator system respectively, thus assuring co-expression of the two genes.</p><p><b>CONCLUSION</b>Successful construction of AAV co-expression system for human angiopoietin 1 and VEGF165 gene will provide the foundation for gene therapy to cure severe ischemic disease.</p>