Humans ; Laparoscopy ; methods ; trends ; Pancreaticoduodenectomy ; methods

OBJECTIVETo design and construct miRNA expression vector dual-targeting on HIF-1α and survivin genes and to investigate its effects on proliferation of human pancreatic cancer cells.

METHODSThe specific pre-miRNA single strand DNA oligos for HIF-1 α and survivin genes were designed and synthesized, then via annealing and ligating with pcDNA6.2-GW/EmGFP-miR plasmids in order, two kinds (eight in total) of miRNA expression vectors for HIF-1α and survivin genes were constructed. The vectors, which were most effective to knockdown target genes, were screened with real-time RT-PCR and combined by chaining technology to construct dual-targeting plasmid. The recombined dual-targeting plasmid, mono-targeting plasmids and negative plasmid were transfected into Panc-1 cells, the suppression effect on two genes was identified by real-time RT-PCR, Western blot and MTT assays.

RESULTSThe miRNA expression plasmids anti-H, anti-S and anti-H+S were successfully constructed by identification of sequencing analysis, and they were able to effectively inhibit the target genes expressing. MTT assays showed that the inhibition effect of dual-targeting vector anti-H+S was higher than that of mono-targeting vectors anti-H and anti-S 72 h after transfection (P<0.05).

CONCLUSIONThe effective miRNA expression vector dual-targeting on HIF-1α and survivin genes has been successfully constructed. The inhibition effect on proliferation of pancreatic cancer Panc-1 cells by dual-targeting plasmid was higher than that by mono-target plasmids.

Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; MicroRNAs ; genetics ; Pancreatic Neoplasms ; genetics ; metabolism ; pathology ; Plasmids ; genetics ; Transfection

BACKGROUNDDaxx has been identified as a nuclear protein that involves in apoptosis and transcriptional repression. Daxx co-localizes with the promyelocytic leukemia (PML) protein and regulates transcription. Human Daxx (hDaxx) is a protein that functions as a transcriptional regulation through its interaction with some DNA-associated proteins. The aim of this study was to explore the transcriptional regulatory effect of hDaxx interacting with adenovirus (Ad) 12 E1B (Ad12E1B) 55-kDa oncoprotein.

METHODSThe co-localization of hDaxx-Ad12E1B or hDaxx-PML protein in the nucleus was observed under a confocal microscope. Interaction of hDaxx and Ad12E1B was analyzed by yeast two-hybrid assay. Direct binding of hDaxx and Ad12E1B was analyzed using coimmunoprecipitation and Western blot in vivo and in vitro. The activity of a luciferase reporter gene, which was regulated by an hDaxx modulated thymidine kinase (TK) promoter, was detected in an automat luminometer.

RESULTSAd12E1B, which co-localized with hDaxx in the nuclei of G401-CC3 cells, disrupted the co-localization of hDaxx and PML in the PML oncogenic domains (PODs). hDaxx bound directly to Ad12E1B in vivo and in vitro. hDaxx interacted with Ad12E1B along its full length. Ad12E1B enhanced transcriptional repression activity of hDaxx.

CONCLUSIONAd12E1B disrupts the co-localization of hDaxx with PML in PODs and enhances transcriptional repression activity of hDaxx.

Adaptor Proteins, Signal Transducing ; Adenovirus E1B Proteins ; physiology ; Carrier Proteins ; analysis ; genetics ; Cell Line, Tumor ; Humans ; Intracellular Signaling Peptides and Proteins ; Neoplasm Proteins ; analysis ; Nuclear Proteins ; analysis ; genetics ; Promyelocytic Leukemia Protein ; Repressor Proteins ; physiology ; Transcription Factors ; analysis ; Transcription, Genetic ; Tumor Suppressor Proteins

Objective To summarize the experience in laparoscopic spleen-preserving distal panereatectomy.Methods From Nov 2003 to July 2006,six patients with distal pancreatic cystic lesions underwent laparoscopic spleen-preserving distal pancreatectomy with splenic vessels preservation. Results All operations were successful with the operative time ranging from 140～265 min and the intraoperative blood loss ranged from 350～600 ml.One case received combination resection of right adrenal adenoma,1 case with combined laparoscopic myomectomy and left ovarian teratomeetamy,1 case with combined laparoscopic myomectomy,1 case with combined laparoscopic cholecysteetomy.All patients were discharged 4 to 9 days postoperatively.The pathologic diagnosis was retained cyst in 2 cases,serous cystadenoma in 2 cases,mucious cystadenoma in 2 cases.Symptoms disappeared in all cases after operations and there was no recurrence during a follow-up period that ranged from 1 month to 31 months.Conclusions Laparoscopic spleen-preserving distal pancreatectomy with splenic vessels preservation is the most suitable procedure for the distal pancreatic benign lesions,and in experienced hands this procedure is safe and effective.

Adult ; Aged ; Female ; Humans ; Intestine, Small ; surgery ; Lymphangiectasis, Intestinal ; pathology ; surgery ; Male ; Middle Aged

OBJECTIVETo evaluate the feasibility and efficacy of laparoscopic distal pancreatectomy.

METHODSTotally 68 patients (male 23, female 45) aged 17 to 77 years, with distal pancreatic lesions, underwent laparoscopic distal pancreatectomy from November 2003 to December 2010. The clinical data were collected. Safety, feasibility and crucial technique manipulation were analyzed retrospectively.

RESULTSAll 68 operations were successful with two cases conversion to open, including 48 cases combined with splenectomy, and 18 cases with preservation of spleen. Fourteen cases received with combination resection of multi-organs, including 4 cases with cholecystectomy, 1 case resection of right adrenal adenoma and cholecystectomy, 1 case with myomectomy and left ovarian teratomectomy; 1 case with right ovarian teratomectomy, 1 case with resection of left adrenal adenoma, 1 case with resection of both adrenal adenoma, 1 case with resection of liver metastasis, 1 case with cholecystectomy and resection of liver metastasis, 1 case with resection of left adrenal adenoma and liver metastasis, 1 case with resection of left adrenal adenoma and colon and spleen, 1 case with biopsy of liver nodule. The mean operative time was (209 ± 58) minutes, the mean intraoperative blood loss was (191 ± 123) ml, and the mean postoperative hospital stay was (8 ± 4) days. The rate of overall postoperative complications was 18.1%, including an 12.1% rate of clinical pancreatic fistula. Only one case needed a reoperation, and there was no postoperative mortality.

CONCLUSIONLaparoscopic distal pancreatectomy with or without splenectomy is safe and feasible in the treatment of most distal pancreatic tumors.

Adolescent ; Adult ; Aged ; Female ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Pancreatectomy ; methods ; Retrospective Studies ; Young Adult

OBJECTIVETo summarize the experience of laparoscopic distal pancreatectomy with preservation of the spleen.

METHODSFrom Nov 2003 to Dec 2004, 2 patients with cystic lesions in the body and tail of the pancreas underwent laparoscopic distal pancreatectomy with preservation of the spleen.

RESULTSBoth of the operations were successful with the operative time of 220 min and 190 min respectively, and the blood loss were 450 ml and 350 ml. There was no postoperative complication and the pathological diagnosis was pancreatic serous cystadenoma. Both patients' symptom disappeared after operation without recurrence during the follow-up of 18 and 5 months.

CONCLUSIONLaparoscopic distal pancreatectomy with preservation of the spleen is safe and feasible for the management of benign tumor in the body and tail of pancreas with the advantages of reduced injury, earlier recovery and less complication.

Adult ; Cystadenoma, Serous ; surgery ; Female ; Follow-Up Studies ; Humans ; Laparoscopy ; Male ; Middle Aged ; Pancreatectomy ; methods ; Pancreatic Neoplasms ; surgery ; Spleen

[Objective]To explore the distribution and expression changes of tight junctional protein JAM-1 in rat models after intracerebral hemorrhage (ICH) and their significance.[Methods]One hundred and twenty-eighty healthy male SD rats were randomly divided into normal control group (n=16) and ICH group (n=112),and the ICH models were induced by stereotactically injecting 75 uL autologous blood into the right caudate nucleus.Seven time points after ICH (6,12,24 and 48 h,and 3,7 and 14 d after ICH,16 rats for each time point) were chosen.BBB permeability was evaluated by Evans blue dye extravasation.The distribution and expression of JAM-1 were detected by immunofluorescence and real-time quantitative PCR.[Results] As compared with that in the normal control group,BBB permeability in the ICH group significantly increased at 24 and 48 h,and 3 and 7 d after ICH (P＜0.05).JAM-1 expression decreased at blood vessels at 12,24 and 48 h after ICH,and JAM-1 expressed at the circulatingleukocytes3 dafterlCH,and abundant JAM-1 positive cells around hematoma were noted in the ED-l-positve macrophages 7 d after ICH.JAM-I mRNA significantly decreased at 12,24 and 48 h after ICH,and significantly increased 7 d after ICH as compared with that in the normal control group (P＜0.05).[Conclusion] JAM-1 experssion changes not only participate in regulation of BBB permeability but also play roles in inflammatory insult after ICH.

Objective To evaluate interventional therapy for biliary stricture (BS) after orthotopic liver transplantation (OLT). Methods The efficacy of interventional therapy for BS after OLT from Oct 2003 to Jan 2006 was analyzed retrospectively. Fifty-three patients received 107 times of interventional therapy through endoscopic retrograde cholangiography ( ERC) which included 68 nasobiliary catheter placements,26 biliary balloon dilatations and stent placements and 13 ERC. Nine patients received 11 times of interventional therapy through percutaneous transhepatic cholangiography ( PTC) including 2 PTC, 7 percutaneous drainages,3 biliary balloon dilatations and 1 biliary stent replacement. One patient received bile drainage through T tube. Results The success rate of ERC was 88. 8% (95/107) , that of nasobiliary catheter placement 94% (64/68) , biliary stent placement 88. 5% (23/26). The success rate of PTC was 81. 8% (9/11) , that of percutaneous drainage was 100% (7/7) , biliary stent replacement 100% (1/1). The curative rate of interventional therapy for 53 patients with BS was 28. 3% (15/53) ,the improvement rate was 41. 5% (22/53). The curative rate of interventional therapy for anastomotic, extrahepatic, intrahepatic hilar and diffuse BS was respectively 66. 7% (4/6)、66. 7% (10/15)、50% (1/2)、0 (0/7) and 0 (0/22). Conclusions The efficacy of interventional therapy for BS after OLT was not satisfactory. The result relates to the type of BS, for anastomotic, extrahepatic and solitary intrahepatic BS this therapy was effective, while that for hilar and diffuse BS the prognosis was poor.

OBJECTIVETo map the frequency and types of EGFR gene mutations present in lung cancer tissues. To evaluate the clinical applicability of a novel real-time double-loop probe PCR of which the ADx-EGFR kit is based, and to compare its performance with traditional Sanger DNA sequencing in the detection of somatic mutations of tumor genes.

METHODSA total of 208 formalin-fixed paraffin-embedded (FFPE) tumor samples were tested. Genomic DNA of the tissue samples was extracted and purified, and subjected to both traditional PCR amplification, Sanger sequencing of EGFR gene in exon 18, 19, 20, 21, and ADx's EGFR mutation detection kit. The mutation rates for EGFR gene in exon 18, 19, 20, 21, as well as the frequency of each mutation detected by the two methods, were analyzed.

RESULTSThe traditional Sanger DNA sequencing technique was successfully performed in 196 out of 208 (94.2%) lung cancer samples, and 22 samples (11.2%) showed EGFR gene mutations. ADx-EGFR kit was successfully used in the lung cancers of all of the 208 cases (100.0%), and 40 samples (19.2%) showed mutations. In the lung cancer samples analyzed, mutations were mainly detected in the exon 19 and exon 21 L858R point mutation, i.e. 4.8% (10/208) and 11.6% (23/208) of total mutations, respectively, and the remaining mutations were rare.

CONCLUSIONSThe success rate of ADx-EGFR real-time PCR for formalin-fixed and paraffin-embedded tissues samples is significantly higher than that of Sanger sequencing (P < 0.01). There are significant differences between the two methods. ADx-EGFR real-time PCR shows a much higher successful detection rate and mutation rate of lung cancer tissues compared with that of Sanger sequencing. As a result, the real-time PCR with ADx-EGFR kit is proved to have a good clinical applicability and a strong advantage over the traditional Sanger DNA sequencing. It is an effective and reliable tool for clinical screening of somatic gene mutations in tumors.

DNA Mutational Analysis ; methods ; Exons ; Genes, erbB-1 ; Humans ; Lung Neoplasms ; genetics ; Paraffin Embedding ; Point Mutation ; Real-Time Polymerase Chain Reaction ; methods