Expression of ZBTB8A in gastric cancer and its clinical significance / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the expression of ZBTB8A (zinc finger and BTB domain containing 8A) in gastric cancer tissues and its clinical significance.</p><p><b>METHODS</b>Level of ZBTB8A mRNA in human normal gastric cell line GES-1, human gastric cancer cell line SGC7901 and MGC803 was detected by real-time PCR. Levels of ZBTB8A mRNA and protein in cancer tissues, adjacent cancer tissues from 104 cases with primary gastric cancer and normal gastric mucosal tissues from 40 cases without malignant gastric diseases were detected by RT-PCR and immunohistochemistry, respectively. Association between ZBTB8A expression and clinicopathology was analyzed.</p><p><b>RESULTS</b>ZBTB8A mRNA expressions in SGC7901, MGC803 and GES-1 cells were 0.00138±0.00015, 0.00158±0.00021, 0.00036±0.000055, respectively, and differences among SGC7901, MGC803 and GES-1 were significant respectively (all P<0.05). ZBTB8A mRNA expression was significantly up-regulated in cancer tissues as compared to adjacent cancer tissues and normal tissues (0.0152±0.0126 vs. 0.0070±0.0061 and 0.0079±0.0036, all P>0.05), while no significant difference was found between adjacent cancer tissues and normal tissues (P>0.05). ZBTB8A expression was significantly associated with invasive depth, lymph node metastasis, tumor stage, and degree of adenocarcinoma differentiation (all P<0.05), but not with age, gender, histological type,gross type (all P>0.05).</p><p><b>CONCLUSION</b>ZBTB8A may be a potential carcinogenic factor in gastric carcinoma, and may also be involved in gastric adenocarcinoma cell differentiation, cancer invasion and metastasis.</p>

[Measurement of activated clotting time during cardiopulmonary bypass in infective endocarditis patients].

OBJECTIVE: To investigate the proper dosage of heparin and protamin in treating the patients with infective endocarditis (IE) during the operation. METHODS: Activated clotting time (ACT) was measured during and after cardiopulmonary bypass (CPB) in 30 patients with IE and 30 patients with rheumatic heart disease (RHD) respectively. RESULTS: The dosage of heparin before bypass in IE group was significantly higher than that of RHD group (P<0.05); the dosage of protamin for neutralization after bypass in IE group was significantly higher than that of RHD group (P<0.05); the ratio of protamin and heparin were 0.76 +/-0.23 in IE group and 0.74 +/-0.12 in RHD group (P>0.05). CONCLUSION: The dosage of protamin should be increased to 3 mg/kg as the heparin is over 400 U/kg before CPB in the patient with IE, but the ratio of protamin and heparin will not be obviously changed.

Transfection of human VEGF165 gene into bone marrow mesenchymal stem cells in rats.

OBJECTIVE: To create a method for transfecting human vascular endothelial growth factor165 (hVEGF165) gene into bone marrow mesenchymal stem cells (MSCs) in rats. METHODS: MSCs of Wistar rats were isolated by density gradient centrifugation and purified based on their ability of adhesion to plastic. Detections of cell surface antigens, including CD34, CD45, CD44, and SH3, were performed using flow cytometry. MSCs' potential of differentiating into osteoblast and lipoblast in vitro was tested. The vector pcDNA(3.1)-hVEGF165 was transfected into MSCs with the liposome mediated method. The expression of hVEGF165 in the transfected cells was detected by enzyme linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (RT-PCR), and Western blot analysis. RESULTS: The cultured MSCs were CD34-, CD45-, CD44+ , and SH+, which were differentiated into osteoblasts and lipocytes successfully. The expressed hVEGF165 in the transfected rat MSCs was demonstrated. CONCLUSION: The vector pcDNA(3.1)-hVEGF165 is successfully expressed in MSCs.

[Effects of myocardial transplantation of mesenchymal stem cells transfected with vascular endothelial factor gene on improvement of heart function and angiogenesis after myocardial infarction: experiment with rats].

UNLABELLED: To establish a method to transfect vascular endothelial factor (VEGF) gene into mesenchymal stem cells ( MSCs) , to investigate the effects of the gene-transfected MSCs on heart function restoration and angiogenesis after myocardial infarction, and to compare the differences among cell therapy, gene therapy, and combined therapy. METHODS: Seventy-one Wistar rats underwent ligation of the left anterior descending coronary artery so as to establish heart ischemia models. Fifteen rats underwent sham operation. MSCs were isolated from several Wistar rats by density gradient centrifugation, purified, and transfected with pcDNA3.1-hVEGF165 or blank plasmid pcDNA3.1 respectively using the liposome mediated method. ELISA, Western blotting, and RT-PCR were used to detect the protein and mRNA expression of hVEG in these MSCs Forty-eight surviving rats that underwent ligation were randomly divided into 4 equal groups: combination group (Combo group) to be injected into the heart infarct zone with suspension of hVEGF165-transfected MSCs 2 weeks after the establishment of the model, cell group to be injected with suspension of MSCs not transfected with VEGF, gene group to be injected with suspension of DNA-liposome containing pcDNA3.1-VEGF165 and control group to be injected with cold culture fluid only. Twelve surviving rats that underwent sham operation were used as normal non-ischemic group. Four weeks after the injection the surviving rats underwent examination of heart functions by the Buxco system. The rats were killed and their hearts were taken out to undergo immunohistochemistry with 5-bromodeoxyuridine (Brdu) and troponin T and factor VIII to measure the area of cardiac infarction and the capillary density. RT-PCR was used to examine the mRNA expression of VEGF. The heart infarcted size was calculated by Evan's blue staining. RESULTS: (1) MSCs can be successfully isolated and cultured by density gradient centrifugation followed by adherence-separation. The expression of hVEGF165 in the transfected MSCs was demonstrated with ELISA, RT-PCR and Western Blot Assay. (2) Four weeks after the cells were transplanted, among all groups but the nonischemic group, the heart infarcted size of the Combo group was 27.8% +/- 3. 0% ,significantly less than those of the cell group (37.0% +/- 10. 1% ) and gene group (37.1% +/- 5.2%, both P <0.05). The heart function of the Combo group was better than those of other groups. (3) The capillary density of the Combo group was 40. 2 +/- 5.5/visual field, significantly greater than those of both the cell group (27.2 +/- 6. 3/visual field, P <0. 01) and that of the control group (18.5 +/- 5.8/visual field, P <0. 01) , and greater to some degree than that of the gene group (35. 8 +/-7.7/visual field, P =0. 189). (4)The heart infarcted size, heart function and capillary density of the cell and gene groups were similar and were smaller, better and greater than those of the control group. (5) Brdu and troponin T double staining detected a varied increase in the number of surviving cardiomyocytes at the heart infarcted area, some of which were double stain positive. RT-PCR showed mRNA expression of hVEGF165 in the Combo and gene groups, that in the Combo group being higher than that in the gene group. CONCLUSION: Eukaryotic expression vector pcDNA3.1-hVEGF165 can effectively be expressed in MSCs. Transplantation of VEGF gene by means of transfected MSCs brings better improvement in myocardial perfusion and in restoration of heart function than either cellular or gene therapy alone.

Low-dose anticoagulation in Chinese patients with mechanical heart valves.

The aim of this study was to provide guidelines for optimal anticoagulation in Chinese patients after mechanical heart valve replacement. A Carbomedics valve was implanted in 178 patients between July 2000 and July 2003. During follow-up, 22 bleeding events and 1 thromboembolic complication occurred. The linearized rates of bleeding and thromboembolism were 5.83% and 0.26% per patient-year, respectively. The linearized mortality rate was 0.79% per patient-year. The final mean international normalized ratio (INR) was 1.68+/-0.38, however there was a significant variation between the early and late periods of follow-up. For Chinese patients with mechanical heart valves, bleeding was the major complication rather than thromboembolism. Low-dose anticoagulation (international normalized ratio 1.4-2.0) could markedly decrease bleeding and effectively prevent thromboembolism. As the INR was most unstable in the first postoperative month, re-examination of patients in this period is critical.

[Anticoagulation in chinese patients with carbomedics mechanical prosthetic heart valves].

OBJECTIVE: To provide some references for defining the Chinese optimal intensity of anticoagulation after mechanical heart valve prostheses replacement. METHODS: For the 178 patients with carbomedics mechanical prosthetic heart valves, the means of INR were compared between the patients with complications and those without complications at the standard of INR1. 4 - 2.0. Also, the variations of INR were compared among different follow-ups. RESULTS: During the follow-up, 22 hemorrhagic and 1 thromboembolic complication occurred. The total linearized rate of anticoagulation-related hemorrhage was 5.83% pty. The total linearized rate thromboembolism was 0.26% pty. The late mortality was 0.79% pty (3 cases ). The final mean INR was 1.68 +/- 0.38. The final mean oral warfarin dose was 2.34 +/- 0.80 mg. The differences of variations of INR in five periods were significant (F = 5.072, P < 0.05). The mean INR in the first month of follow-up was 1.75 +/- 0.27. CONCLUSION: For Chinese patients with mechanical prosthetic heart valve, hemorrhage is the principal complication, the ratio of which is much higher than that of thromboembolism. The low-dose anticoagulation (INR1. 4-2.0) could remarkably decrease hemorrhagic events as effectively as prevent the thrombolic events. Moreover the INR is the most unstable in the first month of follow-up, so re-examination for the patients in the first month after the operation is vitally important.