Adult ; Anastomosis, Surgical ; methods ; Female ; Humans ; Laser-Doppler Flowmetry ; methods ; Male ; Middle Aged ; Surgical Flaps

Anastomosis, Surgical ; methods ; Humans ; Microsurgery ; methods ; Microvessels ; surgery

OBJECTIVETo construct a lentiviral expression vector for RNA interference of human CDH22 gene, and assess its gene silencing effect in colorectal cancer cells to provide a basis for investigating the role of CDH22 gene in the signaling pathway involved in human colorectal carcinoma metastasis.

METHODSHuman CDH22 gene short hairpin RNA (shRNA) sequence was designed using a software available on-line. After synthesis and annealing, the double-stranded oligonucleotides (dsOligoe) were cloned into the pENTR(TM)/U6 plasmid followed by sequence analysis. A positive clone was subcloned into pLenti6/BLOCK-iT(TM)-DEST vector and transformed into stb13 competent cells, with also verification by sequencing. The recombinant lentivirus was harvested from 293FT cells contransfected with the positive recombined plasmid and lentiviral packing materials. SW480 cells were infected with the recombinant lentivirus and the cells with stable CDH22 knock-down were screened by blasticidin selection. CDH22 expression in the cells was determined by real-time reverse transcription-polymerase chain reaction.

RESULTSA recombinant lentiviral vector expressing shRNAs against CDH22 gene was obtained and confirmed by DNA sequencing. Fifteen clones of SW480 cells infected with the recombinant lentivirus were selected, and clone 11 exhibited substantial knock-down of CDH22 mRNA expression.

CONCLUSIONThe lentiviral shRNA expression vector targeting human CDH22 gene capable of stable CDH22 gene knock-down in SW480 cells has been successfully constructed, which provides a basis for further study of the relationship between human colorectal carcinoma and CDH22 gene.

Base Sequence ; Cadherins ; biosynthesis ; genetics ; Cell Line, Tumor ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Molecular Sequence Data ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo identify the region in PRL-3 gene promoter where the transcriptional factor Snail can bind.

METHODSPRL-3 promoter and the possible binding sites of the transcription factors were analyzed by bioinformatical methods. Chromatin immunoprecipitation and PCR were performed using the antibody specific for Snail to verify the binding of Snail to PRL-3 promoter.

RESULTSAccording to the prediction by TRED, a promoter prediction software, PRL-3 gene promoter was located between -700 bp to 299 bp of PRL-3 gene. Many possible transcription factor binding sites such as for Snail, n-MYC, ARNT, E74A, NF-kappaB, NRF-2 and AML-1 were predicted by Consite, a promoter analysis web system. Interestingly, a 5'-CACCTG-3' core sequence and other related sequences of Snail binding sites were found in the promoter region of PRL-3 genes by Consite software. Two regions in PRL-3 promoter were validated to allow binding of Snail by chromatin immunoprecipitation analysis of SW480 cells.

CONCLUSIONSSnail regulates the activity of PRL-3 gene by binding to the promoter of PRL-3 gene in SW480 cells.

Base Sequence ; Binding Sites ; Cell Line, Tumor ; Computational Biology ; Humans ; Molecular Sequence Data ; Neoplasm Proteins ; metabolism ; Promoter Regions, Genetic ; Protein Tyrosine Phosphatases ; metabolism ; Snail Family Transcription Factors ; Software ; Transcription Factors ; metabolism

OBJECTIVETo study the degeneration and regeneration of skeletal muscle after denervation.

METHODSDenervation was carried out in gastrocnemius muscles in 30 adult BALB/C mice by cutting the sciatic nerve. The gastrocnemius muscles were removed at 1, 2, 4, 8, 12, 16 weeks after denervation, respectively. Specimens were processed for histological study and immunohistochemical technique.

RESULTSMuscle fiber atrophy followed by degerneration and regeneration was observed in the early period of denervation. Fusion of the regenerated muscle cells with each other followed by degeneration of the cells and growth of fibro-connective tissue were observed in the later stage. The expression of myoglobin and actin decreased in 1-4 weeks after denervation. The postive expression of the proteins was observed in some 8 weeks' cells and in many degenerated 12-14 weeks' muscle cells.

CONCLUSIONDegeneration and regeneration may coexisted in the denervated muscles. The regenerated muscle cells can't fully develop due to the deficit of nerve regulation and degenerate again. The regenerated muscle cells will melt each other and can't develop to mature muscle fiber in the later stage.

Animals ; Female ; Mice ; Mice, Inbred BALB C ; Muscle Denervation ; Muscle Fibers, Skeletal ; pathology ; Muscle, Skeletal ; innervation ; physiology ; Muscular Atrophy ; physiopathology ; Nerve Regeneration ; physiology ; Sciatic Nerve ; physiology ; surgery

Objective To understand the status on AIDS awareness,AIDS-related behaviors,risk factors on HIV infection status among 15-90 years or older men at the sexually transmitted disease clinics.Methods Data from the 2009 and 2010 national sentinel surveillance system,regarding men attending sexually transmitted disease clinics was collected from Guangdong,Guangxi,Henan,Sichuan,Yunnan and Jiangxi provinces,where the AIDS epidemic among 15-90 years or older population was serious.Data was uploaded to National Center for AIDS/STD Control and Prevention (NCAIDS) through the comprehensive AIDS control and prevention information system.Data was then analyzed by SPSS 18.0.Results A total of 64 003 pieces of data were collected.Among them,8783 ( 13.7％ ) were related to men at ≥ 50 years or older,and to men 15-49 years older were 55 220.The rates on the awareness of AIDS knowledge were from 69.6％ vs.80.1％,on frequently having had commercial sexual contacts in the last three months were between 34.1％vs.36.6％,on having had casual sexual contact in the last three months were 18.7％ vs.28.4％,on having had homosexual anal intercourse as 0.7％ vs.1.4％.The rates of taking HIV antibody testing in the last year (14.3％ vs.17.1％ ) among this population were all significantly lower than the rate among the 15 to 49 years age group.However,the HlV-positive rate among the older age group (fifty years of age or older) was significantly higher than the rate among 15 to 49 year age group ( 1.1％ vs.0.7％ ).Regard the fifty years of age or older men.Data from the multivariate logistic regression analysis demonstrated that factors as:having regular partner (OR=0.588,P=0.034),having homosexual anal intercourse (OR=5.226,P=0.006) were associated with positivities of HIV antibody.Conclusion High-risk sexual behaviors,including homosexual anal intercourse were the major risk factors for men at ≥50 years or older age,related to the infection of HIV.

OBJECTIVETo summarize clinical application experience of treatment of occluded iliofemoral veins with great saphenous vein cross-over bypasses.

METHODSFrom September 1998 to December 2005,6 patients(5 men, 1 woman) with occluded iliofemoral veins underwent great saphenous vein cross-over bypasses. They ranged in age from 36 to 52 years (mean, 41 years old). All patients had unilateral thigh and leg edema and swelling with accompanying pain. The normal contralateral great saphenous vein was dissected and it was tunneled across the super-pubic fat pad and anastomosed to the femoral vein or the proximal portion of great saphenous vein in distal to occluded iliofemoral veins.

RESULTSThe postoperative course was uneventful. One case sustained superficial infection postoperatively at donor site and the gradual wound healed by daily wound dressings. At 1 to 3.5 years (mean 1.8 years) followed up, all patients had pain disappearance and obvious relief of the edema but some swelling.

CONCLUSIONGreat saphenous vein cross-over bypasses is effective operative method of treatment of occluded iliofemoral veins. The operation is relatively simple and safe.

Adult ; Embolism ; surgery ; Female ; Femoral Vein ; surgery ; Humans ; Iliac Vein ; surgery ; Male ; Middle Aged ; Saphenous Vein ; transplantation

OBJECTIVETo summarize clinical application of the sural medial gastrocnemius island muscle flap to cover wound of infection on upper region of the tibial.

METHODSNine patients (7 men, 2 women) with soft tissue defects on the upper region of the tibial underwent reconstruction with the sural medial gastrocnemius island muscle flap. The age ranged from 21 to 60 years old (mean, 34 years). The immediate coverage of the muscle flaps were performed by a meshed split-thickness skin graft. The donor site was closed directly. The donor leg was ipsilateral in all cases.

RESULTSOnly one case sustained superficial infection postoperative and the gradual wound healed by daily wound dressings. All the muscle flaps and skin graft had survived completely without major complication with satisfactory clinical results. All patients were followed-up for 13 months to 4 years (mean 21 months), the donor site was healing, there was no remarkable donor site morbidity.

CONCLUSIONThe sural medial gastrocnemius island muscle flap is nourished by the medial sural artery. The muscle flaps seem to have highly vascularize, a constant vascular anatomy and a long vascular pedicle. The muscle flap is thin and suitable for repairing soft tissue defect on the upper region of the tibial.

Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Muscle, Skeletal ; cytology ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; Tibia ; injuries ; Wound Infection ; surgery

OBJECTIVETo summarize clinical application of the high tibial osteatomy (HTO) with lateral closing-wedge for the treatment of pain of unicompartmental osteoarthritis of the knee.

METHODSFrom February 2000 to February 2004,9 patients (3 males and 6 females, ranging in age from 52 to 58 years, with an average of 56 years) with unicompartmental osteoarthritis of the knee treated by HTO with lateral closing-wedge. The proximal tibiofibular joint was exposed, the anterior part of the capsule was incised, and to remove a laterally based wedge with saw cuts and osteotomes. Stepped staples were used for the fixation of tibial osteotomies after closing the defect of osteotomy.

RESULTSThe operative course was uneventful. There were no complications after operation. The postoperative follow-up period ranged from 2 to 5.5 years (mean, 3.5 years). The results were evaluated with functional assessment criterion of the osteoarthritis of the knee, among the 9 cases, excellent was in 5 cases, good in 3 cases, fair in 1 case.

CONCLUSIONHTO with lateral closing-wedge is an effective operative method for the treatment of pain of unicompartmental osteoarthritis of the knee, but except for older patients over 60 years.

Female ; Humans ; Knee Joint ; surgery ; Male ; Middle Aged ; Osteoarthritis, Knee ; surgery ; Osteotomy ; methods ; Tibia ; surgery ; Treatment Outcome

OBJECTIVETo study proteins correlated with the mechanical properties of engineered cartilage by screening significantly changed proteins during cartilage formation by comparative proteomic analysis.

METHODSHuman chondrocyte, cultured and expanded, were seeded onto a polyglycolic acid/polylactic acid (PGA/PLA) scaffolds. After 4 weeks of culture in vitro, the constructs were divided into three groups. There were 6 specimens in each group. For the regular in vitro culture group (A), the constructs were kept in culture at the original condition for an additional 6 weeks. For in vivo groups, the constructs were implanted subcutaneously into nude mice for either 6 weeks (B) or 12 weeks (C). All specimens were harvested for gross observation, average wet weight and volume measurement, histology, immunohistochemistry and biomechanics to evaluate the results. Meanwhile, comparative proteomic analysis was performed for each group, and those proteins involved in extracellular matrix with at least 2 folds up-regulation were chosen for further exploration. The correlations between Young's modulus and the relative content of the selected proteins were analyzed by Pearson correlation coefficient.

RESULTSAll these samples in the three groups eventually formed hyaline-like cartilage structure. Specimens in C and B groups were similar with adult articular cartilage in appearance, and had multiple mature lacuna in histology. However, those specimens in A group had loose texture with irregular hypertrophy lacuna. Specimens implanted for 12 weeks in vivo had better wet weight (372.5 +/- 35.4) mg and Young's modulus (8.68 +/- 2.65) MPa than those cultured in vivo for 6 weeks (346 +/- 34.5) mg, (3.25 +/- 1.24) MPa (P < 0.01). In group A, they were (184.4 +/- 12.28) mg and (0.7 +/- 0.23) MPa. This study had detected 44 proteins in ECM by comparative proteomic analysis, then chosing the greatest ratio of 6 up-regulation proteins compared between C and A groups. The correlation results indicated the content of Decorin, Chondroadherin and Fibromodulin were linear correlation with the mechanical properties of engineered cartilage (P < 0.05).

CONCLUSIONSComparative proteomic analysis could provide large scale information of associated proteins, making it profit for advanced research on the relationship between extracellular matrix and mechanical properties of engineered cartilage by combination with tissue reconstruction techniques.

Animals ; Cartilage ; cytology ; metabolism ; physiology ; Cells, Cultured ; Chondrocytes ; cytology ; metabolism ; Fetus ; cytology ; Humans ; Mice, Nude ; Proteome ; metabolism ; Proteomics ; Tissue Engineering ; methods ; Tissue Scaffolds