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.
Objective To detect the effects of plasmin combined with hyaluronidase or hexafluoride SF6 on inducing posterior vitreous detachment (PVD). Methods Eighteen young pigmented rabbits were randomly divided into group A, B, and C with 6 rabbits in each. All of the right eyes were the experimental eyes and the left ones was the control. The right eyes in group A, B, and C were injected with plasmin 1 U, plasmin 1 U and hyaluronidase 20 U, and plasmin 1 U and SF6 0.5 ml, respectively; while all of the left eyes underwent intra-vitreous injection with balanced salt solution 0.1 ml. The eyes were observed by indirect ophthalmoscopy, slit lamp examination, biomicroscopy, B-ultrasonography, and electroretinography (ERG) before and after injection respectively. At last, the retinal sections were examined by light microscopy, scanning and transmission electron microscopy. Results The results of scanning microscopy showed incomplete PVD in 2 (33.3%) experimental eyes in group A, and complete PVD in 4 (66.7%) experimental eyes in both group B and C, and the positive rate of PVD in both group B and C significantly differed from that in group A (Plt;0.05). The b-wave amplitudes of ERG in the three groups after injection didn’t differ much from that in the control group or before the injection（Pgt;0.05）. The results of transmission electron microscopy and light microscopy indicated unchanged retinal structure. Conclusions Compared with the application of only plasmin, plasmin combined with hyaluronidase or hexafluoride SF6 can induce complete PVD more efficiently and do no harm to the retina. (Chin J Ocul Fundus Dis, 2005, 21: 388-390)
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.
Objective To determine whether kringle 4-5 could inhibit choroidal neovascularization (CNV) in mice induced by argon laser photocoagulat ion. Methods Fundus laser photocoagulation was performed on C57BL/6J mice to induce CNV. In treatment group, 20 μg (low dosage group) and 50 μg (high dosage group) kringle 4-5 were injected retrobulbarly after photocoagulation. In control group, equilibrium liquid was injected retrobulbarly. Choroidal neovascularization was evaluated on the 7th and 14th day after photocoagulation by fundus fluorescein ang iography. The mice were killed on the 14th day after photocoagulation, the lesions were evaluated histologically and immunohistochemically, and the expression of CD105 was detected. The Expression of VEGF and bFGF was detected by immunohist ochemistry on the 4th day after photocoagulation.Results The incidence of CNV was 64.3% in control group, 51.2%（P＜0.05）in low dosage group, and 44. 1% (P＜0.01) in high dosage group. The CNV lesions were smaller in kringle 4-5 injected eyes in a dose-dependent manner and the number of proliferative vascular endothelial cells in the subretinal membrane of the treated eyes was smaller than that of the control eyes. There was no significant difference of the expression of VEGF and bFGF between the mice in control and treatment group.Conclus ions Kringle 4-5 could inhibit the development of choroidal neovascularization in the experimental mice model.(Chin J Ocul Fundus Dis，2003，19：201-268)
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 explore the clinical value of the soluble urokinase type plasminogen activator receptor (suPAR) level in bronchoalveolar lavage fluid (BALF) for evaluateting the disease severity and prognosis of severe community-acquired pneumonia (SCAP). MethodsEighty-four patients with SCAP were recruited as a SCAP group from the respiratory department, ICU and RICU between April 2014 and April 2016. According to their organ dysfunction, the SCAP patients were subdivided into a MODS group and a non-MODS group. Depending on the treatment response on the 7th day of treatment, they were subdivided into an effective group and an ineffective group. According to the survival condition within 28 days, they were subdivided into a survival group and a death group. Meanwhile, 50 cases with non-severe common community acquired pneumonia were recruited as a control group. On the admission day, all cases were evaluated by PSI score and APACHE Ⅱscore. The serum suPAR level were detected by ELISA on the 1st day in hospital. The suPAR and procalcitonin (PCT) levels in the patient's BALF and serum were detected on the 1st, 3rd, 7th day, discharge or death day. The symptoms and signs, biochemical and pulmonary imaging changes were also observed. ResultsThere were no differences in the sex, age, body weight, duration of pneumonia, or complicated diseases such as hypertension, coronary heart disease and cerebral vascular diseases between the SCAP group and the control group (all P > 0.05). The suPAR levels in serum and BALF of the SCAP group were higher than those of the control group with significant differences (all P < 0.05). The suPAR level in BALF was obviously higher than that in serum in the SCAP group with significant difference (P < 0.05), and slightly higher than that in serum in the control group with no significant difference (P > 0.05). The level of suPAR in BALF of the MODS group was significantly higher than that in the non-MODS group with significant difference (P < 0.05), but there was no significant difference in the PCT level between the two groups (P > 0.05). The suPAR level in the ineffective treatment group was significantly higher than that in the effective treatment group on the 7th day in hospital with significant difference (P < 0.05). The suPAR levels in BALF of the death group were higher than those in the survival group at each time point after admittion with significant difference (all P < 0.05), and the PCT levels had no significant difference between the two groups within 1 week of each time point (all P > 0.05). The suPAR level in BALF of the SCAP group was positively correlated with APACHEⅡ score and PSI score (r=0.578, P=0.0085; r=0.565, P=0.0071), and plasma PCT level was weakly correlated with the APACHEⅡ score and PSI score (r1=-0.0137, r2=-0.0152). ConclusionThe SuPAR level in BALF of patients with SCAP is closely related to the severity and prognosis, and can be used as an index to assess the severity and prognosis.
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 suppressive effect of co mbi nation of tissue plasminogen activator (t-PA), heparin and homoharringtonine on the formation of proliferative vitreoretinopathy (PVR) after vitreoretinal surgery. Methods Forty-three cases (44 eyes )of complicated retinal detachment who received vitreoretinal surgery were divided into 2 g roup s.Twenty cases(20 eyes)in group A were treated by intravitreal injection of abo ve mentioned drugs at the end of operation,while no intraocular injection of drugs given in 23cases(24 eyes)in group B.The mean follow-up period was 7.9 months. Result The rate of recurrent PVR in group A was 15.8%(3 of 19),and 45.5%(10 of 22) in group B (P＜0.05). The rate of recurrent retinal detachment was 5.5%(1 of 18) in group A,an d 33.3%(7 of 21) in group B，in group B(P＜0.05). Conclusion Combination use of the above mentioned drugs can effectively suppress the post operative recurrent PVR and lower the rate of subsequent recurrent retinal detac hment. (Chin J Ocul Fundus Dis, 2001,17:24-25)