Objective: To provide an effective hGM CSF gene transferring vector mediated by gene gun and a basis for study of hGM CSF gene modified tumor cell vaccines. Method: The gastric tumor cell line (SGC) was transfected with eukarytic expression plasmid deoxyribonucleic acid containing the human granulocyte macrophage colony stimulating (hGM CSF) gene using the gene gun. The SGC cell clones (SGC GM CSF 1～5)secreting high hGM CSF level were obtained after G418 resistance selection. The hGM CSF gene had been integrated into chromatosome of SGC by the assay of RT PCR.Results: There was hGM CSF production whose lane was about 30 kD in the culture medium of SGC GM CSF by the assay of SDS PAGE and Western blot. SGC GM CSF had the ability of the high level of GM CSF for a long time(mean 247ng/(10 6 cell?24 h).Conclusion: The hGM CSF gene transferring vector mediated by gene gun was effective and safe. These results provide a basis for study of GM CSF gene therapy for cancer.

Objective:To investigate the clinical significance of thrombospondin-2 (THBS2) mRNA expression in gastric carcino-ma and its relationships with clinicopathologic features, microvessel density (MVD), and matrix metalloproteinase-2 (MMP-2). Meth-ods:THBS2 mRNA expression was detected in 82 cases of gastric carcinomas and adjacent tissues using real-time quantitative fluores-cence polymerase chain reaction. The correlation of this expression with clinicopathologic features was also analyzed. Cluster of differ-entiation 34 (CD34) and MMP-2 protein expression was examined using an immunohistochemical Elivision method. MVD was deter-mined based on CD34-positive tubular structures. Results:The THBS2 mRNA expression level was significantly higher in the gastric carcinomas than in paraneoplastic tissues (P=0.002). The expression was associated with the depth of tumor invasion, MVD, and MMP-2 (P=0.02, r=0.35, P<0.01, and P=0.004, respectively) but not with patient gender, patient age, tumor size, histological type, and lymph node metastasis (P=0.53, P=0.53, P=0.21, P=0.84, and P=0.96, respectively). Conclusions: THBS2 may be significantly in-volved in the occurrence and progression of gastric carcinoma. The effects of THBS2 on gastric tumor growth and metastasis can be monitored by controlling the MMP-2 expression in the carcinoma. However, the specific functions and underlying mechanisms of TH-BS2 require further investigation.

BACKGROUND: Both MRI and transcranial Doppler can be used for ultra-early assessment of acute cerebral infarction (ACI), the former is considered as a reliable diagnostic means mainly for the morphological changes of ACI, however the latter is often used before obvious morphological changes appeared, but their combined diagnostic value is still uncertain. OBJECrIVE: To investigate the value of combined MRI and transcranial Doppler in ultra-early assessment of highly suspected ACI.DESIGN: Case analysis and double-blind design.SETTING: MRI Department of Daqing Oil-field General Hospital and Department of Clinical Epidemiology of West China School, and Medical-Economic Department of Public Health College, Sichuan University.PARTICIPANTS: From May 2000 to July 2001, totally 225 highly suspected ACI patients with stoke-like onset, 121 males and 104 females,aged of 33-86 years old with the means of (62±12), within 12 hours after onset, informed of the experiment, were selected from Daqing Oil-field General Hospital.METHODS: All cases subjected to transcranial Dopller examination and following MRI examination with interval of no more than 1 hour, then received follow-up MRI examination at 15 days later (as golden diagnostic standards for CI). MRI was performed to detect the position, morphology,size and signal of pathological changes. Transcranial Doppler was used to determine the blood flow velocity, peak value of systolic and diastolic stage, frequency spectrum and sound spectrum of blood flow of bilateral cerebral anterior, middle and posterior arteries, internal carotid artery and basal vertebral artery.MAIN OUTCOME MEASURES: The sensitivity, specificity and accuracy in the diagnosis of ACI with MRI, transcranial Doppler and the combination of MRI and transcranial Doppler.RESULTS: Totally 225 cases were selected, and 5 excluded for not accordant to the inclusion standards, finally 220 statistical analyzed. The diagnostic sensitivity of MRI was 79.3%,. specificity of 98.4% and accuracy of 90.5%, comparing to corresponding 80.4%, 81.3% and 80.9% of transcranial Doppler, and 96.7%, 80.5%, 87.3% of the combination of MRI and transcranial Doppler.CONCLUSION: The combination of MRI and transcranial Doppler can be used to make rapid and accurate assessment of ACI due to higher sensitivity and specificity.

BACKGROUND:Tranexamic acid is a synthetic anti-fibrinolytic drug, and can effectively control blood loss after total knee arthroplasty through vein. OBJECTIVE:To evaluate the effectiveness and safety of tranexamic acid in reducing blood loss after primary unilateral total knee arthroplasty. METHODS:From 2014 to 2015, 100 patients from the Department of Lower Limb, Sichuan Orthopaedic Hospital underwent primary unilateral total knee arthroplasty, and randomly divided into test group which used tranexamic acid and control group which used tranexamic acid, with 50 cases in each group. 1 g tranexamic acid was infused into the vein at 10 minutes before total knee arthroplasty in the test group, but tranexamic acid was not given in the control group. RESULTS AND CONCLUSION:The amount of total blood loss, postoperative wound drainage in 24 hours, and the amount of blood transfusion were significantly less in the test group than in the control group (P < 0.05). No significant differences in intraoperative blood loss and the number of patients receiving blood transfusion were found between the test and control groups. No significant difference in preoperative hemoglobin levels was detected before surgery, but hemoglobin levels were decreased after surgery between the two groups. Postoperative hemoglobin levels were apparently higher in the test group than in the control group after surgery. At 24 hours after surgery, no significant difference in coagulation was detectable between the two groups. At 6 days, double lower limb vein color Doppler ultrasound examination did not reveal deep vein thrombosis in both groups. These findings verify that treatment with tranexamic acid in the vein before primary unilateral total knee arthroplasty can evidently diminish the amount of perioperative blood loss and blood transfusion, reduce the risk and cost of blood transfusion, which is good for rehabilitation and does not increase the risk of deep vein thrombosis.

Objective To compare the effects of peripatellar peripheral nerve partially and totally depolarized in total knee arthroplasty (TKA). Methods 66 TKA patients were divided into partially depolarized group (A, 42 knees) and totally depolarized group (B, 38 knees). The scores of knee-joint, Feller and Visual Analogue Scale were recorded. Results There was no significant difference in all the scores between 2 groups before operation (P>0.05). All the scores improved after operation (P<0.05), however, there was still no significant difference between 2 groups (P>0.05). Conclusion Patellar peripheral nerve partially depolarization technique can reduce postoperative knee pain, which has the same effect with totally depolarization.

Objective To explore the clinical curative effect and complications of digital three-dimensional molding of titanium mesh for repairing skull defect. Methods The clinical data of 42 patients having underwent repairing skull defect with three-dimensional molding of titanium mesh were retrospectively analyzed. Results The average operation time was about 2 h, and 42 patients were operated successfully. The bilateral skull was symmetry and the appearance was good. In 42 patients, subcutaneous dropsy occurred in 1 case, epilepsy occurred in 3 cases, and intracranial bleeding again surgery occurred in 1 case. Patients were satisfied with the results of cranioplasty. Conclusions Repairing skull with digital three-dimensional molding of titanium mesh is simple, with shorter operation time, lower operation risk, and lower postoperative complication, and the clinical curative effect is satisfactory.

Objective: To examine the correlation between the promoter methylation of Sterol regulatory-element binding protein-2 (SREBP-2) and miR-33a expression as well as serum markers in patients with coronary artery disease (CAD). Methods: The case-control study. 100 participants who underwent coronary angiography from August 2017 to April 2018 in TaiheHospital, Hubei University of Medicine, were recruited in this study.The methylation level of two fragments, including 12 CpG sites in the promoter region of SREBP-2, have been detected by pyrosequencing in 50 patients with coronary artery disease (CAD) and 50 non-CAD controls. Serum miR-33a level and a panel of 15 CAD related biomarkers were examined by qPCR and routine biochemistry methods. Results: Methylation level of one CpG site (F1-4 loci) in SREBP-2 promoter region were significant higher in CAD patients than in controls(4.56%±0.70% vs 3.54%±0.72%, t=-3.864, P<0.001); methylation level of F1-4 site was negatively correlates with the serum miR-33a levels and high-density lipoprotein cholesterol (HDL-C) levels(r=-0.318, P=0.001; r=-0.225, P=0.024, respectively). Furthermore, F1-4 hypermethylation was an independent risk factor of CAD, independent of age, gender, histories of hypertension, hyperlipidemia, and diabetes(OR=2.452, 95%CI=1.398-4.299, P=0.002). Conclusion These results suggest that DNA methylation and miRNA might cooperate to regulate the lipid metabolism in CAD.

Organoselenium compounds are bioactive substances with extensive physiological activities .As a representa-tive compound ,ebselen could be used as mimics of glutathione peroxidase ,and in the treatment of many diseases ,such as car-diovascular and cerebrovascular diseases ,inflammation and noise-induced hearing loss .the research progress in physiological activities and synthetic methods of ebselen were reviewed in this paper .

OBJECTIVETo investigate the methylation status of Runx3 promoter and Runx3 expression in breast lesion tissues.

METHODSOne hundred and fourteen breast lesions, including 35 cases of fibroadenoma, 39 cases of intraductal carcinoma, 40 cases of invasive ductal carcinoma, and 33 cases of normal breast tissue from Fabruary 2010 to August 2012 were included in this study. Runx3 protein expression was assessed by immunohistochemical SP method; whereas methylation of Runx3 promoter was assessed by high resolution melting (HRM) analysis.

RESULTSRunx3 protein was mainly expressed in the cytoplasm of ductal epithelial cells. The expression rates of Runx3 in normal breast tissue, fibroadenoma, ductal carcinoma in situ, invasive ductal carcinoma were 87.9% (29/33), 85.7% (30/35), 53.8% (21/39), and 40.0% (16/40) respectively. The methylation rates of Runx3 promoter were 12.1% (4/33), 20.0% (7/35), 46.2% (18/39), and 57.5% (23/40), respectively. Correlation analysis between promoter methylation and protein expression of Runx3 in different breast tissue showed the r value in normal breast tissue, fibroadenoma, ductal carcinoma in situ and invasive ductal carcinoma was -0.431 (P = 0.012), -0.408 (P = 0.015), -0.589 (P = 0.000) and -0.743 (P = 0.000) respectively.

CONCLUSIONSRunx3 protein expression shows a downward trend in ductal carcinoma in situ and invasive ductal carcinoma, meanwhile its promoter methylation increases significantly. The methylation of Runx3 promoter may be one of the important factors in the occurrence and development of breast cancer.

Breast ; metabolism ; Breast Neoplasms ; metabolism ; Carcinoma, Ductal, Breast ; metabolism ; Carcinoma, Intraductal, Noninfiltrating ; metabolism ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; DNA Methylation ; Female ; Fibroadenoma ; metabolism ; Humans ; Neoplasm Proteins ; metabolism ; Promoter Regions, Genetic

Objective: To investigate the expression of long non-coding RNA (lncRNA) SNHG8 in EB virus related gastric cancer and their correlation prognosis. Methods: The expression of SNHG8 in 93 gastric cancers and 93 cancer-free controls, matched by age and sex, were determined by real-time PCR. EB virus expression was detected by EBER in situ hybridization. Results: Forty-one gastric cancers were EB virus associated. For all gastric cancers, SNHG8 expression was 14 times higher (P=0.001) than that in non-cancer controls; in the EB virus related gastric cancers, SNHG8 expression was increased 25 times (P<0.05) over EB virus negative gastric cancers. SNHG8 expression level was also significantly associated with TNM staging (P<0.05). Conclusions SNHG8 may act as a proto-oncogene, participating in gastric carcinogenesis.EB virus infection of gastric mucosa may promote SNHG8 expression.