Intravitreal fibrin clots were produced by intravitreal injection of 0.2 ml autologous plasma in 18 rabbit eyes. Subsequently these eyes were treated by intravitreal injection of either tissue plasminogin activator(t-PA) or saline. In the t-PA(12.5mu;g) group(n=10),intravitreal fibrin clots of 6 eyes cleared up within 6 hours, and that of the other 4 eyes within 1 day. In the saline group( n=7), the complete clearallce was not seen after 7 days. The difference of time between two groups was significant ( P＜0. 05), and no evidence of toxic effect was observed as measured by slit-lamp biomicrocopy, ERG, light microcopy and transmission electron microscopy.One eye treated with 100mu;g t-PA also resulted in a total clearance within 6 hours,but retinal toxicity was demonstrated with ophthalmoscopy and light microscopy. （Chin J Ocul Fundus Dis,1994,10:14-16）
Objective To observe the inhibitory effects of local co-transfection of tissuetype plasminogen activator(tPA) gene and proliferating cell nuclear antigen antisense oligodeoxynucleotides(PCNA-ASODN) on the intima proliferation and restenosis of autograft artery in rabbits. Methods One hundred and twenty male Zelanian rabbits were randomly divided into four groups(n=30, in each group): control group, PCNA-ASODN group, tPA group and tPA+PCNAASODN group. The left and right external iliac arteries (length 1.0 cm) were transplanted reciprocally. The transplanted arteries were respectively soaked in lipofection, PCNAASODN, pBudCE4.1/tPA and pBudCE4.1/tPA+PCNA-ASODN solution about 15 minutes. The transplanted arteries were sutured with 9-0 sutures soaked in PCNA-ASODN and pBudCE4.1/tPA solution. Each group were divided into five subgroups(n=6, in each subgroup) according to the sacrifice time (3 d, 7 d, 14 d, 28 d and 56 d after operation). On every sacrifice time point, the vascular specimens were harvested. The thrombocyte assembling and thrombus forming lining vessel wall were observed by scanning electron microscope. The pathological morphology of transplanted arteries were observed under microscope(HE). The intimal areas and stenosis ratio(%) of transplanted arteries were calculate and analyzed statistically among groups by computer system. The mRNA expression of tPA gene in transplanted ressel wall was detected with vevere transcriptionPCR(RT-PCR). The number of PCNA positive cells in transplanted vessel wall was counted by SP immunochemisty.Results The mRNA expression of tPA gene in the transplanted vessel wall in tPA and tPA+PCNA-ASODN groups was higher than that of the other two groups(P＜0.01).The number of PCNA positive cells in the transplanted arteries in PCNAASODN, tPA and tPA+PCNAASODN groups were significantly lower than that of control group(P＜0.05，P＜0.01). The intimal areas and degrees of luminal stenosis of PCNAASODN, tPA and tPA+PCNAASODN groups were lower than those of control group(P＜0.05，P＜0.01), and those of tPA＋ PCNA-ASODN group were lower than those of PCNA-ASODN and tPA groups(P＜0.05). Scanning electron microscopy showed that there were a few thrombocytes lining the vessel wall of tPA group and tPA+PCNAASODN group and no thrombus, whereas there were abundant thrombocytes and thrombi lining the vessel wall of the control group. Conclusion Co-transfection of tPA gene and PCNA-ASODN can effectively inhibit the proliferation of VSMC, hyperplasia of intima and restenosis of transplanted artery.
【Abstract】ObjectiveTo investigate the relationship between the expression of urokinase-type plasminogen activator (uPA) mRNA and breast cancer, lymph node metastasis. MethodsSixty patients with breast tumor were selected randomly and the expression of uPA mRNA was detected with RT-PCR. The patients were divided into benign group (18 cases) and malignant group (42 cases) which included 22 cases with lymph node metastasis and 20 cases without lymph node metastasis. The relationship between uPA mRNA expression and breast cancer, lymph node metastasis was analyzed. ResultsAmong these 18 benign tumors, low expression of uPA mRNA was found in 2 cases and the others were negative. While in 42 cases of malignant tumor, uPA mRNA were positive in 22 cases of lymph node metastasis, 16 of which were high expression, 5 of which were moderate expression, and 1 was low expression. uPA mRNA were positive in 18 of 20 cases of nonmetastatic lymph node, 1 of which was high expression, 5 of which were moderate expression and 12 of which were low expression, the other 2 were negative expression. The expression of uPA mRNA had significant difference between benign and malignant tumors (P＜0.05). The expression in lymph node metastasis was much higher than no lymph node metastasis (P＜0.05). ConclusionThe expression of uPA mRNA in malignant breast cancer is obviously higher than that in benign breast tumor. The expression tensity of uPA is highly relevant to lymph node metastasis in malignant breast cancer, which can provide evidence for clinical staging and therapy.
Objective To investigate the therapeutic effects of throm bolytic drug infusion via carotid artery on experimental central retinal artery occlusion (CRAO), and observe the changes of fibrinolytic activity in the system ic circulation. Methods To dissolve the thrombi in 15 cats (30 eyes) with CRAO established by laser irradiating a branch of central retinal a rtery after intravenous injection of photochemical drugs, urokinase (UK) was dir ectly infused via carotid artery in 5 cats (10 eyes) in group A or intravenously injected in 5 cats (10 eyes) in group B, and isotonic saline solution was intra venously injected in 5 cats (10 eyes) in group C respectively. The patency of the artery was evaluated by fundus fluorescein angiography. Moreover, the changes of fibrinolitic activity in the blood were observed by blood biochemical examination. Results Four hours after UK infusion, the complete repatency proportion was 80% (5 cats 8 eyes) in group A, and 50% (4 cats 5 eyes) in group B. There was significant difference between the two groups. Besides, after the infusion, the indexes of coagulation, fibrinolysis, and anti-fibrinolysis in group A were better than those in group B and C (Plt;0.01). Conclusion In the treatment of experimental CRAO, thrombolytic drug infusion via carotid artery is better and more effective than via intravenous injection, which may provide a new method of thrombolytic drug delivery and animal models. (Chin J Ocul Fundus Dis，2004，20：186-188)
ObjectiveTo investigate the therapeutic effect of kinetin on bleomycin A5 (BLM-A5)-induced pulmonary fibrosis in rats. MethodsSixty female Wistar rats were randomly divided into three groups. Group A (n=20) was intratracheally injected with saline as control. Group B (n=20) were intratracheally injected with BLM-A5 to establish pulmonary fibrosis model. Group C (n=20) was intratracheally injected with BLM-A5 and received intraperitoneal injection of kinetin at 0.5 mL/100 g once daily. The rats were sacrificed on the 3rd,7th,14th and 28th day respectively. HE and Masson staining were performed to observe lung pathological changes. The contents of hydroxyproline (HYP),urokinase-type plasminogen activator (u-PA),tissue-type plasminogen activator (t-PA),and PAI-1 in lung and plasma were measured by ELISA. ResultsAlveolitis was most obvious on the 7th day and pulmonary fibrosis was most severe on the 28th day in group B compared with other two groups (P<0.05). Alveolitis and pulmonary fibrosis in group C were significantly alleviated compared with group B (P<0.05),but still more severe than group A (P<0.05). The HYP contents in group B,coincided with fibrosis,began to increase on the 7th day and reached the peak on the 28th day,significantly higher than those in other two groups (P<0.05). The u-PA contents of lung tissue in group B began to decline on the 3rd day,reached the minimum on the 7th day,and was still significantly lower than those in other two groups (P<0.05).On the 14th day, the u-PA contents had no significant difference among three groups. The u-PA plasma contents in group B began to decline on the 3rd day,reached the minimum and had significant difference compared with other two groups on the 7th day (P<0.05),and there was no significantly difference among three groups after the 14th day. The t-PA contents change of lung tissue and plasma in three groups were generally consistent with u-PA,but the t-PA plasma contents in group B were still significantly lower than those in group A on the 14th day (P<0.05). The PAI-1 contents of lung tissue in group B began to increase on the 3rd day,reached the maximum on the 7th day,was still significantly higher than those in other two groups (P<0.05),and there was no significant difference among three groups on the 14th day. The PAI-1 contents in group C decreased compared with those in group B (P<0.05),but still higher than those in group A (P<0.05),and there was no difference among them on the 14th day. The PAI-1 plasma contents in group B began to increase on the 3rd day,reached the maximum and was significantly higher than other two groups on the 7th day (P<0.05),and there was no significant difference among three groups on the 14th day. ConclusionThe contents of u-PA and t-PA are increased by inhibiting PAI-1 generation in lung tissue through kinetin treatment,so that,kinetin can suppress pulmonary fibrosis induced by BLM-A5.
Abstract: Objective To investigate the messenger ribonucleic acid (mRNA) expression level of tissue-type plasminogen activator (t-PA) in endothelial cells derived from adult mesenchymal stem cells (MSCs) after fluid shear stress loading which is within the physiological range. Methods After culturing in vitro, bone marrow MSCs of SD rats were seeded on slides.When it come to 80% confluence,26 slides were exposed to 5dyn/cm2 fluid shear stress for 3h in a flow chamber, and then induced to endothelial cells. Among them,13 slides constituted group Ⅰ, and the rest 13 slides set up group Ⅱ, which would be cultured for 3-4d further and passaged in 1∶3. At the same time, control group was set up, which including the cells never exposed to fluid shear stress before the endothelial differentiation. Fluid shear stress were exerting to cells in a specially made flow chamber. The expression level of t-PA mRNA of all groups were measured by real-time fluorescent quantitation reverse transcriptionpolymerase chain reaction (RTPCR). Results After endothelial differentiation for 7 days, the SD rats bone marrow MSCs acquired typical endothelial cell appearance. The t-PA mRNA expression level of group Ⅰ and group Ⅱ have an obviously enhance compared with control group(P＜0.05). The t-PA mRNA expression level of group Ⅱ step down a little (P＞0.05), but it is still significantly higher than that of control group (P＜0.05). Conclusion Fluid shear stress could provide a protective action on the endothelial cells induced from MSCs in vitro, and the effect maintains with the cells passages. This formulates a theoretical foundation to the therapeutics of atherosclerosis and selection of seed cells in vascular tissue engineering.
ObjectiveTo observe the efficacy and safety of vitrectomy combined with submacular injection of tissue-type plasminogen activator (t-PA), gas filling and anti-VEGF drugs (multiple therapy) for thick submacular hemorrhage.MethodsA retrospective case study. From January 2014 to June 2018, 24 patients (24 eyes) with thick submacular hemorrhage who received multiple therapy at the Department of Ophthalmology of Peking University Third Hospital were included in the study. Among them, there were 15 males and 9 females with the average age of 69.05±8.86 years. The average submacular bleeding time was 17.15±10.30 days, the average bleeding area was 13.85±8.63 DD. Seventeen eyes showed hemorrhagic RPE detachment. The international standard visual acuity chart was used to BCVA examination, which was converted to logMAR visual acuity in statistics. The frequency domain OCT was used to measure the height of the foveal elevation. The average logMAR BCVA of the affected eye was 1.37±0.38. The average height of the macular fovea was 824.94±294.38 μm. All eyes underwent 23G or 25G vitrectomy. During the operation, 0.1-0.5 ml t-PA (10 μg/0.1 ml) was injected under the macula. The vitreous cavity was filled with 15% C3F8 after gas-liquid exchange in 13 eyes, and 11 eyes were filled with sterilized air. Eleven eyes were injected with anti-VEGF drugs at the end of the operation, and anti-VEGF drugs were given PRN treatment according to the activity of the lesion. The average follow-up time after treatment was 27.90±22.21 months. The absorption of bleeding under the macula, the improvement of vision, the occurrence of rebleeding and treatment-related complications were observed and recorded. The Wilcoxon rank sum test was performed to compare the BCVA and the height of foveal elevation before and after treatment.ResultsOne month after the treatment, the blood in the fovea of all affected eyes was cleared. At the last follow-up, the logMAR BCVA and macular foveal elevation were 0.82±0.28 and 253.88±71.75 μm, respectively. Compared with those before treatment, the difference was statistically significant (Z=－3.727, －3.234; P＜0.001, ＜0.001). The average intravitreal injection of anti-VEGF drugs was 1.08 times. During the operation, a tiny hole was formed in the center of the macula when t-PA was injected under the retina. Two eyes showed mild vitreous hemorrhage early after the operation. During the follow-up period, bleeding recurred in 2 eyes.ConclusionsVitrectomy combined with submacular injection of t-PA, gas filling, and anti-VEGF drugs is an optimal combination for the treatment of thick submacular hemorrhage. It can effectively remove submacular hemorrhage, improve vision, reduce foveal elevation with good safety.
Objective To observe the clinical effect of intravenous thrombolytic therapy for central retinal artery occlusion (CRAO) with poor effect after the treatment of arterial thrombolytic therapy. Methods Twenty-four CRAO patients (24 eyes) with poor effect after the treatment of arterial thrombolytic therapy were enrolled in this study. There were 11 males and 13 females. The age was ranged from 35 to 80 years, with the mean age of (56.7±15.6) years. There were 11 right eyes and 13 left eyes. The visual acuity was tested by standard visual acuity chart. The arm-retinal circulation time (A-Rct) and the filling time of retinal artery and its branches (FT) were detected by fluorescein fundus angiography (FFA). The visual acuity was ranged from light sensation to 0.5, with the average of 0.04±0.012. The A-Rct was ranged from 18.0 s to 35.0 s, with the mean of (29.7±5.8) s. The FT was ranged from 4.0 s to 16.0 s, with the mean of (12.9±2.3) s. All patients were treated with urokinase intravenous thrombolytic therapy. The dosage of urokinase was 3000 U/kg, 2 times/d, adding 250 ml of 0.9% sodium chloride intravenous drip, 2 times between 8 - 10 h, and continuous treatment of FFA after 5 days. Comparative analysis was performed on the visual acuity of the patients before and after treatment, and the changes of A-Rct and FT. Results After intravenous thrombolytic therapy, the A-Rct was ranged from 16.0 s to 34.0 s, with the mean of (22.4±5.5) s. Among 24 eyes, the A-Rct was 27.0 - 34.0 s in 4 eyes (16.67%), 18.0 - 26.0 s in 11 eyes (45.83%); 16.0 - 17.0 s in 9 eyes (37.50%). The FT was ranged from 2.4 s to 16.0 s, with the mean of (7.4±2.6) s. Compared with before intravenous thrombolytic therapy, the A-Rct was shortened by 7.3 s and the FT was shortened by 5.5 s with the significant differences (χ2=24.6, 24.9; P＜0.01). After intravenous thrombolytic therapy, the visual acuity was ranged from light sensation to 0.6, with the average of 0.08±0.011. There were 1 eye with vision of light perception (4.17%), 8 eyes with hand movement/20 cm (33.33%), 11 eyes with 0.02 - 0.05 (45.83%), 2 eyes with 0.1 - 0.2 (8.33%), 1 eye with 0.5 (4.17%) and 1 eye with 0.6 (4.17%). The visual acuity was improved in 19 eyes (79.17%). The difference of visual acuity before and after intravenous thrombolytic therapy was significant (χ2=7.99, P＜0.05). There was no local and systemic adverse effects during and after treatment. Conclusion Intravenous thrombolytic therapy for CRAO with poor effect after the treatment of arterial thrombolytic therapy can further improve the circulation of retinal artery and visual acuity.
The activities of tissue- type plasminogen activator (t-PA) and plasminogen activator inhibitor(PAl) in plasm from 61 patients with retinal vein occlusion (RVO) were measured by using chromatogenous substrate s-2390 assay.The results showed that the t-PA activity in the patients with RVO were decreased (1.69plusmn;0. 56IU/ml, P＜0.01) and PAI activity increased (8.80plusmn;1.60AU/ml, P＜0. 01) comparing with health subjects 2.07plusmn;0.40IU/ml and 7.33plusmn;0.67AU/ml respectively. Among the patients, t-PA activity in the patients with ischemic retinopathy was more obviously decreased (1.35plusmn;0.43IU/ml, P＜0.01) and the activity of PAI was increased (9.35plusmn;1.37AU/mi) comparing with those patients suffering from nonischemic retinopathy (the activities of t-PA and PAI were 1.92 + 0.53IU/ml and 8.42plusmn;1.29AU/ml respectively, Plt;0.01). In addition, these changes were getting more obvious with the degree of severity of the disease. These results indicated that there was disorder in the balance between t-PA and PAI in patients with RVO,which my play an important role in the course of occurrence and development of RVO, especially in ischemic type. （Chin J Ocul Fundus Dis,1994,10:71-73）
ObjectiveTo investigate the therapeutic effects of thrombolysis infusion via microcatheter on the treatment of central retinal artery occlusion(CRAO). MethodsUrokinase (UK) was directly infused via ophthalmic artery (OA) by microcatheter (6 patients) or via intravenous (7 patients) to dissolve the thrombus. The patency of the artery was evaluated by fundus fluorescein angiography (FFA), and the effect of fibrinolytic activity on the systemic changes was observed by blood biochemical examination simultaneously. ResultsIn 6 patients in the microcatheter group, 5 had completely and 1 had partly reopened OA on the morrow of UK infusion with the patency rate of 83.33%, while in 7 patients in vein group, 3 completely reopened, 2 partly reopened and 2 obstructed OA were found with the patency rate of 42.86%. The difference between the two groups was significant. No obvious change of index of blood coagulation system was found in catheter group, which had great disparity compared with the vein group.ConclusionUrokinase infusion via microcatheter in CRAO has better therapeutic impact and smaller effect on systemic action. (Chin J Ocul Fundus Dis, 2005,21:16-19)