ObjectiveTo investigate the effects of thrombospondin-1 active fragment (TSP-1) synthetical peptide VR-10 on proliferation and migration of rhesus choroidal-retinal endothelial (RF/6A) cell and the expressions of apoptosis relative genes in RF/6A cell.
MethodsThe survival rate of RF/6A cell were detected by methyl thiazolyl tetrazolium, and migration ability was measured by transwell chamber after exposure to 1.0 μg/ml TSP-1 and synthetic peptide VR-10 (0.1, 1.0, 10.0 μg/ml) for different times (6, 12, 24, 48 hours). Caspase-3 and factor associated suicide (FAS) protein levels were measured by Western blot. The mRNA level of bcl-2 and FAS ligand (FASL) were measured by reverse transcription-polymerase chain reaction (RT-PCR).
ResultsThe survival rate of RF/6A cells was determined by the treatment time and concentration of TSP-1(1.0 μg/ml) and the synthetic peptide VR-10 (0.1, 1.0, 10.0 μg/ml). The lowest survival ratio of RF/6A was 78% (P < 0.001) when cells were treated by 10 μg/ml synthetic peptide VR-10 after 48 hours. TSP-1 and synthetic peptide VR-10 could inhibit migration of RF/6A cells in transwell chamber (P < 0.001). 10.0 μg/ml synthetic peptide VR-10 had the strongest effect, 1.0 μg/ml TSP-1 was the next. Migration inhibition rate was increase with the increase of the concentration of VR-10 (P < 0.001). There was no significant differences between 0.1 μg/ml and 1.0 μg/ml VR-10 (P=0.114). Western bolt showed that RF/6A cell in control group mainly expressed the 32×103 procaspase-3 forms. To 10.0 μg/ml VR-10 treated group, it showed decreased expression of procaspase-3 (32×103) and concomitant increased expression of its shorter proapoptotic forms (20×103). Compared with control group, expression of FAS peptides were significantly increased in 10.0 μg/ml VR-10 treated group. Compared with control group, expression of FasL mRNA was significantly increased in 10.0 μg/ml VR-10 treated group(t=39.365, P=0.001), but the expression of bcl-2 mRNA was decreased(t=-67.419, P=0.000).
ConclusionTSP-1 and synthetic peptide VR-10 had the ability to inhibit proliferation and migration of endothelial cell, and also induce apoptosis by increasing FAS/FASL expression and repressing bcl-2 expression.
Objective To explore the feasibility of high-pressure injection to transfer human thrombomodulin (hTM) gene into arterial wall of rabbits.Methods Eighty-four healthy New Zealand rabbits were randomly divided into three groups: pcDNA3.1/hTM plasmid group (n=28), pcDNA3.1(+)/neo plasmid group (n=28) and untransfected group (n=28). After gene transfection, the model of arterial injury-blocking was established. Then, the expressions of hTM mRNA and protein in arterial wall were examined by RT-PCR and immunohistochemistry at 3 d, 7 d, 14 d and 28 d after operation. Results Seventeen rabbits died accidentally from the day of operation to 3 d after operation. The expressions of hTM mRNA of different time points in pcDNA3.1/hTM plasmid group were significantly higher than that in pcDNA3.1(+)/neo plasmid group and untransfected group (Plt;0.01). For the expressions of hTM mRNA at different time points in pcDNA3.1(+)/neo plasmid group and untransfected group, the difference of inter-group and intra-group was not significant (Pgt;0.05). hTM protein was expressed in every group and mainly localized in the inner lining of arterial wall. The expressions of hTM protein at different time points in pcDNA3.1/hTM plasmid group were significantly higher than that in pcDNA3.1(+)/neo plasmid group and untransfected group (Plt;0.05). The expression of hTM protein at different time points in pcDNA3.1(+)/neo plasmid group and untransfected group kept relative constancy, the difference of inter-group and intra-group was also not significant (Pgt;0.05). Conclusion High-pressure injection is feasible to transfer pcDNA3.1/hTM plasmid into arterial wall of live animals.
ObjectiveTo explore the ultrasonic morphological indexes influencing the effectiveness of ultrasound-guided thrombin injection (UGTI) in the treatment of iatrogenic femoral artey pseudoaneury (IFAP).MethodsThe patients with IFAP treated by UGTI in the West China Hospital of Sichuan University from 2015 to 2017 were collected according to the inclusion criteria. The demographic data, comorbidities, clinical symptoms, ultrasonic morphological parameters, thrombin dosage, and postoperative complications were analyzed. The risk factors of technical success of UGTI were analyzed by logistic regression analysis. The technical success was defined as absence of flow within the FAP immediately after UGTI. ResultsA total of 53 patients who met the criteria of inclusion were included in this study. The technical and treatment success rates of UGTI were 84.9% (45/53) and 96.2% (51/53), respectively. No deep venous thrombosis, arterial thromboembolism, infection, bleeding, allergy, and other complications or death occurred in all patients. There were no statistical differences in the age, clinical symptoms, comorbidities (except hypertension, P=0.040), ultrasonic morphological parameters (except femoral artery diameter, P<0.001), and thrombin dosage between the patients with technical success and failure (P>0.05). Multivariate logistic regression analysis showed that the smaller femoral artery diameter was the risk factor of the technical failure [OR=0.350, 95%CI (0.165, 0.743), P=0.006]. Further adjustment of thebaseline data of patients (excluding patients with hypertension), the logistic regression analysis still found that smaller femoral artery diameter was the risk factor of the technical failure [OR=0.419, 95%CI (0.205, 0.860), P=0.018].ConclusionsUGTI in treatment of IFAP is minimally invasive and has a higher technical success rate. Smaller femoral artery diameter might affect technical success rate of UGTI.
ObjectiveTo explore the value of modified method for intratumoral injection of thrombin in the treatment of iatrogenic pseudoaneurysms.MethodsClinical data of 28 patients with iatrogenic pseudoaneurysms after interventional treatment in our hospital from October 2012 to June 2018 were retrospectively analyzed. Twenty-one cases were treated with ultrasound-guided thrombin injection for pseudoaneurysms (Ultrasound group), and seven cases were treated with DSA-mediated balloon occlusion and thrombin injection for pseudoaneurysms (DSA group). The patients were followed-up at 1 day, 1 month and 3 monthS after operation.ResultsThe total success rates of the two groups were 100%. There was no treatment-related complications in the two groups. There was no recurrence after 1–3 months of treatment.ConclusionsIntratumoral injection of thrombin can be used for the treatment of iatrogenic pseudoaneurysm. The effect of the improved treatment is more significant. These two methods can be used as the best way to treat iatrogenic pseudoaneurysm with different neck diameters.
Objective To investigate whether the sleep-induced hypoxemia ( SIH) at different time and different level have different effects on pulmonary emphysema and coagulation systemfunction in the rats with pulmonary emphysema. Methods Thirty Wistar rats were randomly divided into three groups( n = 10 in each group) . All rats were exposed to cigarette smoke twice a day ( 30 min each time) . From29th day on, the rats in Group A ( pulmonary emphysema with short SIH) were also exposed to mixed gas of 12. 5% oxygen for 1. 5 hours during sleeping time every day ( the expose time was divided into 4 periods, 22. 5 min each) . The rats in Group B ( pulmonary emphysema with mild SIH) were also exposed to mixed gas of 15% oxygen for three hours during sleeping time every day( the expose time was divided into 4 periods, 45 min each) . The rats in Group C( pulmonary emphysema with standard SIH) were also exposed to mixed gas of 12. 5% oxygen for three hours during sleeping time every day( the expose time was divided into 4 periods,45 min each) . After continuous exposure for 56 days, the rats were sacrificed. Semi-quantitative image analytic method was employed for histopathological analysis including pathological score of lungs, mean linear intercept ( MLI) and mean alveolus number( MAN) . ATⅢ, FIB, vWF, FⅧ were measured. Results All animals in three groups manifested the histopathological features of emphysema. Pathological scores of lungs and MLI of every group were significantly different from each other( F = 21. 907, F = 18. 415, all P lt; 0. 05) , Group A [ ( 61. 90 ±4. 25) % , ( 92. 45 ±1. 78) μm] and Group B[ ( 64. 60 ±3. 95) % , ( 92. 80 ±3. 65) μm] were significantly lower than Group C[ ( 73. 30 ±3. 86) % , ( 99. 32 ±2. 81) μm, q= 8. 96, q =6. 84, q = 12. 64, q =9. 65, all P lt; 0. 05] . Levels of FIB were significantly different among three groups ( F = 20. 592, P lt; 0. 05) while FIB in Group A[ ( 189. 98 ±5. 29) mg/ dL] and Group B[ ( 182. 70 ±2. 78) mg /dL] were significantly lower than that in Group C[ ( 198. 40 ±7. 37) mg/ dL, q = 4. 86, q= 9. 07, all P lt; 0. 05] , and FIB in Group A was significantly higher than that in Group B( q = 4. 20, P lt; 0. 05) . Levels of FⅧ were significantly different from each other( F = 33. 652, P lt;0. 05) while FⅧ in Group A[ ( 232. 26 ±4. 17) % ]and Group B[ ( 242. 53 ±14. 50) % ] were significantly lower than that in Group C[ ( 303. 25 ±32. 93) % ,q= 10. 73, q = 9. 18, all P lt; 0. 05] . Conclusions Pulmonary emphysema and hypercoagulable states increases with time and severity of SIH in rats with pulmonary emphysema. The elevated activity of blood coagulation factor may be a critical role in the hypercoagulable states.
ObjectiveTo explore the efficacy of thrombin in treatment of subcutaneous effusion after radical resection of breast cancer.
MethodsOne hundred and ninety patients underwent radical resection of breast cancer from July 2008 to July 2013 in this hospital were divided into postoperative observation group and postoperative control group according to the operation time. A daily injection of thrombin by drainage tube was performed on day 3 after operation in the postoperative observation group, the negative pressure drainage only was performed in the postoperative control group. The drainage volume in 72 h after operation, time of extubation, cases of subcutaneous effusion were counted after operation. Then the patients with subcutaneous effusion were divided into subcutaneous effusion observation group and subcutaneous effusion control group according to the time of extubation, the thrombin was injected into cavity after pumping subcutaneous effusion with pressing and dressing in the subcutaneous effusion observation group and only pressed after pumping subcutaneous effusion in the subcutaneous effusion control group, respectively. The healing time of subcutaneous effusion was counted in these two groups.
ResultsCompared with the postoperative control group, the drainage volume in 72 h after operation was less(P < 0.001), the time of extubation was earlier(P < 0.001), the rate of subcutaneous effusion was lower(P < 0.05), color of drainage fluid on day 2 after mastectomy was lighter(P < 0.001)in the postoperative observation group. Compared with subcutaneous effusion control group, when subcutaneous effusion was 20-50 mL or > 50 mL, the healing time of subcutaneous effusion was significantly shorter in the subcutaneous effusion observation group(P < 0.05).
ConclusionsInjecting thrombin by drainage tube after operation can reduce the drainage volume, decrease the rate of subcutaneous liquid, and shorten the time of extubation. Injecting thrombin into cavity of subcutaneous liquid can shorten the healing time of patients with middle and large subcutaneous effusions after radical resection of breast cancer.
