Objective To construct the bicistronic retroviral vectors containing GFP gene and human survivin gene(wildtype and T34A mutant). Methods Survivin fragments were amplified from eukaryotic expression vectors by PCR, then cloned into pMIG retroviral vectors. The constructed recombinant was identified by double digestion with restriction enzymes. The constructed retroviral vectors was transfected into Phoenix E package cells under mediation of liposome. The expression of mRNA was examined by RT-PCR and the transcription of gene was determined by FACS analysis of GFP. Results The target fragments were successfully bound to pMIG vector. After the vectors were transfected into NIH3T3 cells, the expression of mRNA was confirmed by RT-PCR and the expression of GFP was confirmed by FACS analysis. Conclusion Human survivin gene wildtype and T34A mutant retroviral vectors have been constructed successfully, which can provide strong molecular tools for further studies and a novel approach for selective cancer gene therapy.

Air Pollutants, Occupational ; analysis ; Benzenesulfonates ; analysis ; Chromatography, High Pressure Liquid ; methods ; Workplace

Objective To analyze the incidence of remaining symptoms after discectomy with different operation,and investigate its causes and clinical significance.Methods Two hundred and twentyfour patients were followed up.The incidence of remaining symptoms and clinicai situation after discectomy were observed with different operation:discectomy (110 patients,single group),discectomy with bone grafting internal fixation (70 patients,fusion group),and discectomy with dynamic fixation (44 patients,non-fusion group).The SPSS 19.0 software was used to analyze the data.Results One hundred and sixpatients (47.32％,106/224) occurred remaining symptoms in 224 patients.In single group was 69 patients (62.73％,69/110),in fusion group was 26 patients (37.14％,26/70),and in non-fusion group was 11 patients (25.00％,11/44).The incidence of remaining symptoms among three groups was significant difference(x2 =22.177,P =0.000).The incidence of remaining symptoms in single group was significantly higher than that in non-fusion group and fusion group (x2 =17.921,P =0.000;x2 =11.235,P =0.001).The incidence of remaining symptoms in non-fusion group and fusion group had no significant difference (x2 =1.817,P =0.178).Conclusions There is significantly different in the incidence of lumbar disc herniation with different surgery methods.Discectomy with bone grafting internal fixation and with dynamic fixation can effectively reduce the incidence of remaining symptoms.

Arteriovenous Malformations ; therapy ; Child ; Female ; Humans ; Pulmonary Artery ; abnormalities ; Pulmonary Veins ; abnormalities ; Septal Occluder Device

Objective To investigate the biological effect of 50 Hz sinusoidal electromagnetic fields at 1 mT on the proliferation of the human osteosarcoma cell line MG-63.Methods Osteosarcrma MG-63 cells were divided into control and experimental groups.The control group was incubated without an electromagnetic field; the experimental group was incubated in a 50 Hz,1.0 mT sinusoidal electromagnetic field.On the 2nd,4th and 6th day,their proliferation was determined using a cell counting kit-8(CCK-8)assay.Variations in the cell cycle were detected with flow cytometry(FCM).Semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR)was used to measure cyclin B1 and cyclin D1 mRNA.Results Compared with the control group,proliferation of the experimental group cells was reduced significantly.The percentage of cells at G0-G1 phase increased,and the mRNA expression of cyclin B1 and cyclin D1 was significantly reduced.Conclusions A 50 Hz sinusoidal electromagnetic field at 1.0 mT can inhibit the proliferation of osteosarcoma cell line MG-63 significantly.

An α-amylase inhibitor (α-AI) was isolated from white kidney beans (Phaseolus vulgaris. L) by ethanol fractional precipitation, ion exchange chromatography and gel filtration column chromatography. It was a homogeneity glycoprotein demonstrated by SDS-PAGE and gel filtration on CL-6B. The glycoprotein contained 88.2% protein and was rich in aspartic acid, glutamic acid, leucine, threonine and serine. The carbohydrate moiety was consisted of Man, Glc, Gal and Xyl in a mole ratio of 2.42∶1.50∶1.52∶1.00. The glycan and the core protein backbone was connected by O-linkage as determined by β-elimination reaction. The continuous oral administration of the α-AI (150 mg·kg-1·d-1 ) for 7 days can lower fasting blood glucose and 300 mg·kg-1 ·d-1 α-AI for 7 days can improve the sugar tolerance on alloxan-dependent diabetic model rats. The result showed the α-AI obtained from white kidney beans had good hypoglycemic effect on alloxan induced diabetic rats and may have high potential pharmaceutical value as a regulative digestive-starch degradation in patients suffering from diabetes.

Objective To assess the efficacy of laryngeal mask airway (LMA) i-gel in patients required insertion of nasogastric tube before laparoscopic surgery. Methods Fifty-five ASA Ⅰ - Ⅲ patients ( Mallampati Ⅰ -Ⅲ ), aged 26-64 yr, weighing 54-73 kg, scheduled for elective laparoscopic surgery were randomly divided into 2 groups: group Ⅰ ( n= 28) and group Ⅱ ( n = 27). The size of i-gel LMA was chosen based on the patient' s weight, and i-gel LMA was inserted after induction of anesthesia with TCI of propofol and remifentanil, and iv injection of rocuronium. In group Ⅰ , the nasogastric tube was inserted through the drain tube of i-gel LMA. In group Ⅱ , the nasogastric tube was inserted through the nostril before operation. The hemodynamic parameters, SpO2 ,PETCO2 and peak airway pressure were monitored during operation. The fiberoptic laryngoscopy scores were assessed and nasogastrice tube displacement was recorled after successful LMA placement. The LMA placement time, success rate of LMA placement at the first attempt, airway sealing pressure, the occurrence of air leakage of LMA, and nasogastric tube drainage were recorded. The problems after removal of the LMA were observed and the adverse reactions within 24 h after operation were recorded. Results The hemodynamics was stable and the SpO2,peak airway pressure were within the normal range during operation in both groups. There was no significant difference in the LMA placement time, success rate of LMA placement at the first attempt, nasogastric tube drainage rate, airway sealing pressure, incidence of air leakage, fiberoptic laryngoscopy scores, problems after removal of the LMA and adverse reactions between the two groups ( P ＞ 0. 05). Conclusion i-gel LMA can provide adequate ventilation and does not interfere with the nasogastric tube drainage during laparoscopic surgery and can be used effectively for the patients required insertion of nasogastric tube before operation.

Objective To introduce a new classification system for femoral intertrochanteric fractures and propose classification to assist surgical management.Methods A total of 176 cases of femoral intertrochanteric fractures treated from September 2007 to October 2011 were enrolled in the study.X-ray and three-dimensional spiral CT reconstruction were performed to define fracture shape,location,number of major fragments and displaced direction.As combined with the intraoperative findings and cadaveric anatomical study,the femoral intertrochateric fractures was simulated using a three-dimensional entity of normal proximal femur produced by Mimics 11.1 program.A new classification system for femoral intertrochateric fractures was proposed.Results The new system categorized intertrochateric fractures of the femur into five types (Type Ⅰ,Type Ⅱ,Type Ⅲ,Type Ⅳ and Type Ⅴ) based on the integrity or destruction of the medial column,lateral column and posterior column set and divided on the basis of bone connection of femoral head-neck fracture fragments with the bone in intertrochanteric region.Conclusions The new classification system with introduction of three-column concept is brief and practical and has a clear definition of fracture stability.Besides,the new classification-based treatment is recommended.

Objective To detect the expression and function of ZNRF3 in different kinds of thyroid cancer tissues and cells.Methods Immunohistochemistry was used to detect the expressions of ZNRF3 protein in 35 cases of papillary thyroid carcinoma and 10 cases of poorly differentiated thyroid carcinoma.The expressions of ZNRF3 gene in TPC-1 and 8505C were detected by RT-PCR,and the cell lines were derived from papillary thyroid carcinoma and poorly differentiated thyroid carcinoma respectively.After silenced ZNRF3 gene expression with lentivirus,the proliferation ability of TPC-1 cells were detected with CCK-8,the invasion and metastasis ability of TPC-1 cells were detected with Transwell.Results According to results of immunohistochemistry and RT-PCR,the expressions of ZNRF3 in papillary thyroid carcinoma cells and tissues were higher than those in poorly differentiated thyroid carcinoma cells and tissues,the differences were statistically significant (4.83±0.44 vs.3.13 ±0.59,t =2.20,P ＜0.05;1.01±0.06 vs.0.21±0.04,t =11.80,P＜0.01).After ZNRF3 geng silencing,according to the results of CCK8,the proliferation ability of TPC-1 cells was significantly enhanced in 72 h,the difference was statistically significant (0.96 ± 0.10 vs.0.64 ± 0.05,t =3.19,P ＜ 0.05);and according to the results of Transwell,the TPC-1 cell's invasion (0.12 ± 0.01 vs.0.09 ±0.00,t =5.48,P＜0.01) and migration (0.22 ±-0.01 vs.0.17 ±0.01,t =4.58,P ＜0.05) also increased,the differences were statistically significant.Conclusion The expression of ZNRF3 in papillary thyroid carcinoma is higher than that in poorly differentiated thyroid cancer.ZNRF3 is tumor suppressor gene in the thyroid tumors.

Objective:To investigate the effects of electroacupuncture (EA) at Neiguan (PC 6) and Baihui (GV 20) by observing the changes of CCAAT/enhancer-binding protein(C/EBP) homologous protein (CHOP) and caspase-12 gene expressions in rats after cerebral ischemia-reperfusion injury (IRI), and explore whether the apoptosis pathway of endoplasmic reticulum stress (ERS) is involved in the protective mechanisms of EA.
Methods:Sixty rats were randomly assigned to five groups (12 in each group): a normal control group (group A), a sham-operation group (group B), an operation group (group C), an Edaravone group (group D) and an EA group (group E). The cerebral IRI rat model was induced by middle cerebral artery occlusion (MCAO) using intraluminal monofilament. 2,3,5-triphenyl tetrazolium chloride (TTC) staining was adopted in the measurement of cerebral infarction volume. Real-time polymerase chain reaction (RT-PCR) was used to determine the mRNA expressions of CHOP and caspase-12.
Results: Compared with group A and group B, the volume of cerebral infarction and mRNA expressions of CHOP and caspase-12 in group C, group D and group E were increased, with statistical significances (P<0.05 orP<0.01); compared with group C, the volume of cerebral infarction and mRNA expressions of CHOP and caspase-12 in group D and group E were decreased significantly (P<0.05 orP<0.01); there were no significant differences between group D and group E in comparing the above items (P>0.05).
Conclusion:EA at Neiguan (PC 6) and Baihui (GV 20) can effectively suppress the volume of cerebral infarction. Furthermore, the underlying mechanism of EA at Neiguan (PC 6) and Baihui (GV 20) is possibly related to the down-regulation of CHOP and caspase-12 mRNA expressions, so as to decrease cell apoptosis.