Objective To explore the feasibility of repairing the large area skin and soft tissue defect in thefront upper of tibia by free flap with anastomosis of saphenous vessels.Methods From June,2009 to April,2014,16 cases (10 males and 6 females) of large area skin and soft tissue defect in the front upper of tibia were treatedwith free flap.The ages ranged from 24 to 56 years old,averaged of 34.5 years old.The supportive therapy and repeated debridement combined with VSD,and designed to use free anterolateral thigh flap with anastomosis of saphenous vessels before repairing operation.Results Fourteen patients accorded with preoperative design,in which 2 cases were adopted cross leg flap anastomosis posterior tibia vessels of the healthy side (1 case of saphenous artery diameter too small,and 1 case of saphenous artery long injury degeneration,unsuitable for vascular anastomosis).All 14 flaps survived,expect 2 cases were part-necrosis in the end of the flap,and gradually healed by dressing exchange.Conclusion Reconstruction of large area skin and soft tissue defect in the front upper of tibia with free flap can use anastomosis of saphenous vessels.

BACKGROUND AND OBJECTIVEAs a barrier to metastases, cells normally undergo apoptosis after they lose contact with their extra cellular matrix (ECM). This process has been termed "anoikis". Tumour cells that acquire malignant potential have developed mechanisms to resist anoikis and thereby survive after detachment from their primary site while traveling through the lymphatic and circulatory systems. This "anoikis resistance" is considered the first step to tumor metastases. The aim of this study was to screen metastasis-associated genes from anoikis resistant and adherent growth A549 lung cancer cell by Human Genome Array.

METHODSEstablish anoikis resistant A549 lung cancer cell lines by using poly-hydroxyethyl methacrylate resin processed petri dishes, which causes cell free from adherent. The different expressed gene between anoikis resistant A549 cell and adherent growth A549 cell was tested using human V2.0 whole-genome oligonucleotide microarray, a product of Capitalbio Corporation, Beijing. Screen metastasis-associated genes.

RESULTS745 different expressed genes were screened, including 63 highly metastasis-associated genes.

CONCLUSIONThe successfully established anoikis resistant A549 cell lines and screened different expressed genes provide us basis for further research on metastasis of lung cancer.

Anoikis ; genetics ; physiology ; Cell Line, Tumor ; Flow Cytometry ; Gene Expression Profiling ; Genome, Human ; genetics ; Humans ; Lung Neoplasms ; genetics ; Oligonucleotide Array Sequence Analysis

BACKGROUNDTo observe the suppressive effects of exogenous p27 gene on human lung cancer cell line A549.

METHODSAn adenovirus expression vector (pAd CMV-p27) containing 570 bp human full-length p27 cDNA was transfected into human lung cancer cell line A549. Expression of exogenous p27 gene was detected by dot-blot hybridization and laser co-focal system. MTT was adopted to measure the effects of exogenous p27 gene on cell cycle progression and cell features of the infected A549.

RESULTSThe mRNA and protein expression level of p27 was remarkably increased after transfecting with exogenous p27 gene. The apoptosis of infected A549 occurred and the progression of cell cycle was arrested in G1 phase.

CONCLUSIONSp27(kipl) gene transfer may play a therapeutic role in the treatment of lung cancer.

BACKGROUNDTo investigate the biological effects of anti VEGF₁₆₅ ribozyme on human lung adenocarcinoma cell.

METHODSHammerhead ribozyme (VRz) against VEGF₁₆₅ gene transcripts (site 212) and its paired mutant ribozyme (mVRz) were designed and synthesized, and the cleavage activity of the ribozymes on target RNA in a cell-free system was observed. The replication-incompetent adenovirus-mediated eukaryotic expression vectors (rpAdVRz) containing VRz and mVRz gene were constructed and identified. Then the human lung adenocarcinoma cells (A549) were infected with recombinant adenovirus. The biological characteristics of A549 cell before and after infection in vitro were inspected by Northern blot, laser confocal imaging system analysis, flow cytometry and transmission electron microscopy.

RESULTSVRz specifically and efficiently cleaved the VEGF₁₆₅ mRNA. The rpAdVRz was successfully constructed and infected A549 cell. The level of VEGF₁₆₅ expression decreased 87% in rpAdVRz infected cells compared with the other groups, but their biological characteristics were not influenced by the expression of the exogenous gene.

CONCLUSIONSThe adenovirus mediated hammerhead ribozyme against VEGF₁₆₅ can significantly decrease the expression of VEGF₁₆₅. This provides an experimental basis for human lung cancer gene therapy with antiangiogenesis method.

OBJECTIVE: To detect the mutation site in a pedigree affected with autosomal dominant polycystic kidney disease (ADPKD) and verify its impact on the protein function. METHODS: Peripheral blood samples were collected from the proband and his pedigree members for the extraction of genomic DNA. Mutational analysis was performed on the proband through whole-exome sequencing. Suspected variant was verified by Sanger sequencing. A series of molecular methods including PCR amplification, restriction enzyme digestion, ligation and transformation were also used to construct wild-type and mutant eukaryotic expression vectors of the PKD2 gene, which were transfected into HEK293T and HeLa cells for the observation of protein expression and cell localization. RESULTS: The proband was found to harbor a c.2051dupA (p. Tyr684Ter) frame shift mutation of the PKD2 gene, which caused repeat of the 2051st nucleotide of its cDNA sequence and a truncated protein. Immunofluorescence experiment showed that the localization of the mutant protein within the cell was altered compared with the wild-type, which may be due to deletion of the C-terminus of the PKD2 gene. CONCLUSION The c.2051dupA (p. Tyr684Ter) mutation of the PKD2 gene probably underlay the pathogenesis of ADPKD in this pedigree.
DNA Mutational Analysis ; Female ; Frameshift Mutation ; HEK293 Cells ; HeLa Cells ; Humans ; Male ; Pedigree ; Polycystic Kidney, Autosomal Dominant/physiopathology* ; Protein Kinases/genetics* ; Protein Transport/genetics* ; Whole Exome Sequencing

OBJECTIVE: To explore the genetic basis for a Chinese patient with amyotrophic lateral sclerosis (ALS). METHODS: Peripheral blood samples were collected from the patient and his parents for the extraction of genomic DNA. Genetic variant was identified by whole exome sequencing. Candidate variant was verified by Sanger sequencing of his parents and healthy controls. RESULTS: The patient was found to harbor a heterozygous c.420C>G (p.Asn140Lys) variant of the SOD1 gene. The same variant was not detected in his parents and 100 healthy controls. The variant has not been included in HGMD, dbSNP and other databases. CONCLUSION The c.420C>G variant of the SOD1 gene may underlie the ALS in this patient. Above finding has enriched the spectrum of SOD1 gene variants.
Amyotrophic Lateral Sclerosis/genetics* ; China ; Heterozygote ; Humans ; Superoxide Dismutase-1/genetics* ; Whole Exome Sequencing

Objective To study the clinical effect of diversion treatment for complex anal fistula.Methods A total of 60 patients with complex anal fistula were enrolled in this study were divided into control group and experimental group according to the random number table method,with 30 cases per group.All patients were given routinely imaging examination and other related checks,and the intestine tract was cleaned in the morning of the operative day.The control group were treated with low anal fistula resection,while the experimental group with diversion treatment.The efficacy of treatment,postoperative anal function,wound healing time and pain were compared between the two groups.Results The total effective rate in the experimental group was higher than that in the control group (P ＜ 0.05).The Wexner score of the anus function in the experimental group was lower than that in the control group at 1,7,14 and 21 d after the operation,and the differences were statistically significant (P ＜ 0.05).The wound healing time,VAS score on 14thpostoperative day,intraoperative wound area and postoperative scar size were lower in the experimental group than that in the control group (P ＜ 0.05).Conclusion Diversion treatment for complex anal fistula has significant efficacy,faster postoperative wound healing,and it can effectively relieve clinical symptoms and signs,improve anal function,reduce body pain,so it is worthy of clinical promotion.

Objective To study the clinical effect of diversion treatment for complex anal fistula.Methods A total of 60 patients with complex anal fistula were enrolled in this study were divided into control group and experimental group according to the random number table method,with 30 cases per group.All patients were given routinely imaging examination and other related checks,and the intestine tract was cleaned in the morning of the operative day.The control group were treated with low anal fistula resection,while the experimental group with diversion treatment.The efficacy of treatment,postoperative anal function,wound healing time and pain were compared between the two groups.Results The total effective rate in the experimental group was higher than that in the control group (P ＜ 0.05).The Wexner score of the anus function in the experimental group was lower than that in the control group at 1,7,14 and 21 d after the operation,and the differences were statistically significant (P ＜ 0.05).The wound healing time,VAS score on 14thpostoperative day,intraoperative wound area and postoperative scar size were lower in the experimental group than that in the control group (P ＜ 0.05).Conclusion Diversion treatment for complex anal fistula has significant efficacy,faster postoperative wound healing,and it can effectively relieve clinical symptoms and signs,improve anal function,reduce body pain,so it is worthy of clinical promotion.