Objective To discuss the endothelial cell which was modified by exogenous anticoagulant genes contribute to the increase of antithrombosis activity of lined vascular prosthesis and the influence to other physiological functions of endothelial cells. Methods This summarized paper was made on literature review of recent years.Results The transfection of genes, including plasminogen activator (t-PA, u-PA, Urokinase), thrombomoduline (TM) and hirudin, etc, to endothelial cells resulted in not only the increase of antithrombosis activity of local vascular, but also the decrease of endothelial cell function in adherence and proliferation.Conclusion The increase of antithrombosis activity of lined vascular prosthesis has been done by exogenous genes. However, this technique ought to be studied, intensively.

Objective To elucidate the mechanism of accelerated apoptosis of monocytes in patients with lupus nephritis (LN). Methods 17 LN patients were enrolled in the study and 15 healthy volunteers were selected as controls. The expression of Faa and Fas ligand (FasL) was examined by immunofluorescent staining and flow cytometer analysis. The apoptosis and survival rate of monocytes cultured in vitro with human FasL (rHu-FasL) was analyzed by flow cytometer analysis and 5-diphenyl tetrazolium bromide (MTT) staining, respectively. Results Faa expression on monocytes was significantly higher in LN patients than that in healthy subjects. There was no difference of monocytes Fas expression between active LN patients and silent LN patients. There was no detectable FasL expression on monocytes from both LN patients and healthy controls. When monocytes cultured in vitro with rHu-FasL, there was a significantly increasing apoptosis and a decreasing survival rate of monocytes in LN patients as compared to healthy controls. Conclusion The up-regulated expression of functional Fas on monocytes may contribute to the accelerated apoptosis of monocytes in LN patients.

The incidence of locally recurrent rectal cancer is about 10%. Most patients have serious symptoms after tumor recurrence, which seriously affects the quality of life and prognosis of patients. Although it is more difficult to perform reoperation, it is still the main method to treat local recurrence of rectal cancer. At present, the main methods of reoperation include transabdominal anterior resection, combined pelvic organ resection, combined pelvic organ resection, and sacral resection. Open surgery is the main method. In recent years, with the extensive development of laparoscopic technology, laparoscopic techniques have been tried at home and abroad to perform reoperations for locally recurrent rectal cancer, showing good short-term results. The authors systematically introduce the application of laparoscopic technology in the reoperation of locally recurrent rectal cancer based on relevant research advances at home and abroad, in order to explore its clinical application prospects and promotion value.

Objective To evaluate the safety and effect of local resection of the duodenum for the treatment of gastrointestinal stromal tumors(GISTs). Methods In this study, 12 duodenal GIST cases in which tumors located more than 1.0 cm away from the major duodenal papilla, underwent simple local resection of the duodenum along with the tumors measuring at 4 cm in diameter or less. The clinical data were retrospectively analyzed. Results After simple local resection of the duodenum for GISTs, no stomal leak occurred and the margin was all negative. Pathologically tumors were all low risk grade. There was no recurrence during a follow-up of 9-38 months. Conclusions Local resection of the duodenum is safe and efficient in the treatment of duodenal GISTs should the tumors locat more than 1.0 cm away from the papilla.

Objective To construct and identify the recombinant vector pcDNA3. 1 (-) B/myc-BRMS 1 carrying breast-cancer metastasis suppressor 1 (BRMS 1) which can express in eukaryote cells and which will provide the basis for further researching the mechanisms of metastasis suppression and working on cancer metastasis gene ther-apy. Methods To isolate total RNA from MCF - 7 cells and design a pair of primers, and coding sequence of aRMS 1 cDNA were amplified from human breast cancer cells MCF -7 by reverse transcription-polymerase chain reaction (RT-PCR). Then the product was inserted to the PcDNA3. 1/myc-His (-) B plasmid. The recombined pcDNA3. 1 (-)B/myc-BRMS1 was identified by gene sequence analysis,then recombinants was transfected into HEK-293 cells and was identified by Western blot. Results The recombinant of pcDNA3.1 (-) B/myc-BRMS1 was structurally confirmed by analysis of sequencing. The inserted fragment in the vector was in the right direction and its sequence was structurally confirmed to be consistent with CDS sequence of human BRMSI cDNA that of the published data. GenBank, [AF159141]. The recombinants was transfected into HEK-293 cells ,then the cells expressed protein tagged c-myc identified by Western blot indicated it can express in eukaryote cells. Conclusion cDNA of human BRMS1 can be successfully cloned and inserted into Eukaryote-expression vector. The newly constructed vector may serve as the potential tool to conduct further comprehensive experiments in future on BRMS1 function and on gene therapy.

AIM: To provide experimental evidence for gene therapy of thrombophilia disease, we constructed the eukaryotic expression plasmid with human thrombomodulin (hTM) gene and observed the alteration of hTM expression on the surface of human umbilical vein endothelial cells (HUVECs) with and without the reconstructive plasmid. METHODS: The whole expressive fragment of hTM gene was amplified by PCR from human genome. Both hTM gene and pcDNA3.1(+)/neo empty vector was digested by HindⅢ and EcoRⅠ. Two digested fragments were ligated into pcDNA3.1/hTM with T_4DNA ligase. After identifying, the reconstructive plasmid transfected into HUVECs using lipofectin. The hTM antigen on the HUVECs was detected by immunohistochemistry. RESULTS: The hTM reconstructive plasmid was confirmed by double endonuclease redigesting and sequencing. About 10% HUVECs were transfected by pcDNA3.1/hTM plasmid with lipofectin and the high-level hTM was detected on the transfected cells. CONCLUSION: We constructed the pcDNA3.1/hTM plasmid successfully, and it could be expressed on the HUVECs. [

Objective To retrospectively analyze whether the kyphosis exists after removal of the internal fixators for thoracolumabar vertebrae fractures. Methods A total of 18 patients (35-68 years old) with thoracolumabar vertebrae fractures (T11-L2 ) were fixed with short segment pedical screw. The fixators were removed one year postoperatively to observe the changes of the Cobb' s angle and trauma vertebra'height. Results All the patients were followed up for 6-24 months ( average 18.7 months),which showed no intraoperative or postoperative complication, breakage or loosening of the screws. Compared to the Cobb angle and the vertebra height before removal of the internal fixators, the average loss of the Cobb angle was 0.7° and that of the vertebra height was 0.8 mm six months after removal of the fixators, 1.9° and 1.1 mm respectively one year after removal of the fixators, and 2.4° and 1.3 mm respectively two years after removal of the fixators in 16 patients without osteoporosis (P ＞0. 05). Among two patients with osteoporosis, the average loss of the Cobb angle and the vertebra height was 6° and 8°respectively and 3 mm and 5 mm respectively six months after removal of the fixators; 13° and 17° respectively and 5 mm and 7 mm respectively one year after removal of the fixators; 15° and 19° respectively and 6 mm and 7.5 mm two years after removal of the fixators. Conclusions After the internal fixation for thoracolumbar vertebrae burst fractures, kyphosis develops mildly, with insignificant change of the vertebral height. While the kyphosis becomes worse after removal of the fixators for thoracolumbar vertebrae burst fractures in patients with osteoporosis.

Objective To investigate The Th1/Th2 and Tc1/Tc2 polarization in the peripheral blood of first degree relatives of pemphigus vulgaris(PV) and healthy control individuals, and to approach the mechanism of the Dsg3-specific autoimmunity in PV. Methods The peripheral blood mononuclear cells (PBMC) from first degree relatives and healthy control was stimulated for72 h with Dsg3 and without Dsg3. Th1/Th2, Tc1/Tc2 was assessed by four-color flow cytometry. Results The mean frequency of Dsg3-spe-cific Th2 cells for PV antibody positive first degree relatives was 10.13%±3.72%, compared with stimula-tion without antigen 7.28%±3.58%, the difference was significant (P<0.05). The percentage Dsg3-spe-cific Th2 was markedly higher in the PV antibody positive first degree relatives group than that in the control group(10.13%±3.72% vs 6.10%±2.82%, P<0.05) , Tc2 was markedly higher also (20.01%± 10.43% v514.91%±8.06%, 20.01%±10.43% vs 9.58%±5.49%, P<0.05). Conclusion When Dsg3 stimulated PBMC were used to stimulate autologous T cells an increased amount of Th2 and Tc2 was observed, it is implied that the imbalance of Th1/Th2, Tc1/Tc2 might play an important role in the initia-tion of PV.

Objective To investigate therapeutic autcomes of using telomerase inhibitors to treat cancer at the presumably most and least opportune circadian stages basing on our earlier study. Methods Twenty-four BALB/C nude mice were synchronized to a regimen of LD12:12 for 4 wk. Hepatic cancer cells (SMMC-7721) were implanted into both flanks of each mouse.Two weeks after transplantation,the hTERT-5'RZ was used to treat the hepatic cancer transplanted into the nude mice daily for two weeks,the injection times being either 9 or 21 HALO.Results The tumorinhibition ratio of mice treated at 21 HALO (65%) was statistically significantly higher than that of mice treated at 9 HALO (48%). Telomerase activity was also reduced to a greater extent in mice treated withhTERT-5'RZ at 21 than at 9 HALO, that was at the time of maximal circadian telomerase activity. Conclusion Injection of ribozyme targeted to telomerase during the tumor's DNA synthesis is associated with a betterinhibition of tumor growth and a better therapeutic outcome in hepaticcancer.

Objective To explore the significance of MYCN gene amplification in children with neuroblastic tumors(NT).Methods The clinicopathological data of 154 cases with NT were reviewed,including general data, classification of pathology,clinical stage and prognosis.MYCN gene amplification was detected by fluorescence in situ hybridization(FISH) and its relationship between pathological characteristics and prognostic significance was analyzed.Results There was 154 cases of NT aged 1 day to 11 years,with a mean age of 26.1 months,and the median age of 20.5 months.Male and female ratio was 1.48 : 1.00.According to International Neuroblastoma Staging System (INSS) ,20 cases were of stage Ⅰ (13.0％) ,23 cases of stage Ⅱ (14.9％) ,43 cases of stage Ⅲ (27.9％) ,64 cases of stage Ⅳ(41.6％) and 4 cases of Ⅳs (2.6％).There were 72 cases(46.8％) with favorable histology,and 82 cases(53.2％) with unfavorable histology.MYCN amplification was found in 20 cases (13.0％) and the signal ratio of MYCN and chromosome 2 (CEP2) was 4.08-43.29.One hundred and thirty-four cases of MYCN non-amplification included MYCN gain in 91 cases(68.0％) ,MYCN negative in 43 cases(32.0％).MYCN expression showed the significant differences in ages, neuroblastoma type, international neuroblastoma pathology classification (INPC), mitosis karyorrhexis index (MKI), and clinical stages (all P ＜ 0.05).No significant difference was found in gender(P ＞ 0.05).Of 20 MYCN amplification cases,4 cases (20.0％) survived and 16 cases (80.0％) died,and the overall survival rate was 20.0％ (4/20 cases) ,with survival time was (17.10 ± 2.24) months;of 134 MYCN non-amplification cases,96 cases (71.6％) survived and 38 cases (28.4％) died, with survival time of (28.71 ± 1.28)months.Survival analysis showed the cases with MYCN amplification had worse prognosis (x2 =19.596, P ＜ 0.05).Conclusions Patients with MYCN amplification had poorer prognosis and lower incidence of MYCN amplification of pediatric NT was found in China.