Objective To explore the effects of Zhaoke defibrase and anti alpha;vbeta;3mAb (23C6) on the adhesion and immigration of bovine retinal vascular endothelial cells. Methods The culture dishes coated with vitronectin (Vn) and collagen，assays of adhesion and immigration were performed 60 minutes after different concentration of Zhaoke defibrase and anti-alpha;vbeta;3 mAb was added to the bovine retinal vascular endothelial cells. The apoptosis of bovine retinal vascular endothelial cells induced by Zhaoke defibrase and anti-alpha;vbeta;3 mAb was detected by electron microscopy. Results Both Zhaoke defibrase and anti-alpha;vbeta;3 mAb inhibited the adhesion and immigration of bovine retinal vascular endothelial cells in a dose-dependent manner. The inhibited concentration (IC50) of Zhaoke defibrase was less than 0.05 mu;mol/L, while (IC50) of anti-alpha;vbeta;3 mAb was more than 2.5 mu;mol/L. 81.8% endothelial cells adhering to Vn were inhibited by 0.1 mu;mol/L Zhaoke defibrase, while 76.3% by endothelial cells adhering to Vn were inhibited by 10 mu;mol/L anti-alpha;vbeta;3 mAb. Typical apoptosis cells were found in bovine retinal vascular endothelial cells after affected by Zhaoke defibrase and anti-alpha;vbeta;3 mAb. Conclusion Both Zhaoke defibrase and anti- alpha;vbeta;3mAb can significantly inhibit the adhesion and immigration of bovine retinal vascular endothelial cells to extracellular matrix， and the mechanism may lie in inducing the apoptosis of endothelial cells. (Chin J Ocul Fundus Dis, 2005,21:118-121)
Abstract： Objective To observe the significance of the changes of cell adhesion molecules (CAM) CD11b/CD18 and sPselectin during the perioperative period of open heart surgery under cardiopulmonary bypass (CPB), and investigate the roles of CD11b/CD18 and sPselectin in systemic inflammatory response triggered by CPB. Methods Thirty patients including 18 males and 12 females, age ranged from 29 to 55 years (45.3±8.1 years) having undergone valvular replacement for rheumatic heart disease in our hospital were selected as the subjects of this research. After anesthesia induction, radial arterial blood sample was collected at six different time points including the time prior to skin incision, and 30 min, 1 h, 6 h, 12 h and 24 h following the start of CPB. The expression levels of CD11b/CD18 were tested by flow cytometry, and concentration of sP-selectin in the plasma was measured with enzymelinked immunosorbent assay(ELISA). Results The expression of CD11b/CD18 was elevated at 30min after CPB, and it reached the peak (581.44±215.26) at 6 h after CPB with significant differences (Plt;0.05). Its expression started to drop at 12 h after CPB, but it was still higher than the expression level before CPB. The expression returned under the level before CPB at 24 h after CPB with insignificance differences (Pgt;0.05). The expression of sPselectin in the peripheral blood started to rise evidently at 30 min after CPB, reaching the peak (51.44±10.06 ng/ml) with significant differences (Plt;0.05). Its expression level decreased at 12 h after CPB and fell back below the level before CPB with insignificant differences (Pgt;0.05). Conclusion CPB can cause the expression of CD11b/CD18 and sPselectin to rise in the peripheral blood, which may play an important role in the systemic inflammatory response triggered by CPB.
OBJECTIVE To evaluate the effects of sodium hyaluronate on adhesion prevention after flexor tendon surgery. METHODS In 47 cases with the flexor tendon surgery, two kinds of sodium hyaluronate jelly preparations were injected into the tendon sheath before suture. Preparation I (20 mg/2 ml) was for group A (17 patients) and preparation II (20 mg/2 ml) was for group B (16 patients). The control group (group C, 14 patients) were treated in the same way except injection of sodium hyaluronate. The functions of afflicted fingers including flexibility, pain and swelling were measured immediately, at the first, second and the third month after operation. RESULTS All 47 patients were followed up 1 to 3 months. 64.71% patients in group A and 68.75% in group B showed significant improvement. There were significant difference compared with group C (P lt; 0.05). There were no significant adverse reactions were observed in all groups. CONCLUSION Two sodium hyaluronate preparations have effects in adhesion prevention after flexor tendon surgery with safety and expedience.
Objective To investigate the clinical effect of chitosan in prevention of knee dysfunction due to adhesion after operation for patellar fracture. Methods From March to October 1999, 40 cases of patellar fracturewere treated by internal fixation, with intraarticular injection of 2% chitosan in only 24 cases after fixation and with no chitosan injection in 16 cases(control group). The function of the knee joint, including extension and flexion, was evaluated 1month and 1 year after operation respectively. Results One month after operation, the knees with chitosan injection could actively move in the average range of 104°±23°, and the knees in the control group could move in the average range of72°±16°, which showed significant difference between two groups(P＜0.01); 1 year after operation, the range of movement of the knees with injection was 165°±38° on average, and that of the knees in the control group was 110°± 31°, which also indicated significant difference between two groups (P＜0.05). Conclusion Medical chitosan could effectively prevent or reduce the post-operative adhesion of knee joint after patellar operation.
Objective To investigate the effects of human acellularamnion membrane on SD rat tendon adhesion and to obtain the experimental data for clinical application in preventing postoperative tendon adhesion. Methods The tendons of 28 adult SD rats hindlimb were cut and sutured. The tendons of left hindlimb were encapsulated by human accellular amnion membraneas the experimental group and the ones of the other side were not encapsulatedas control group. The rats were killed 1, 2, 4, 6, 8 and 12 weeks after operation. The results were evaluated grossly and histologically. Results There were no differences in healing of injury tendon and inflammatory response between the two groups. The anatomical and histological results showed the experimental group had less adhesion than the control group(Plt;0.05). Conclusion Human acellular amnion membrane can prevent adhesion of tendonwithout affecting tendon healing and is an optimal biological material to prevent tendon adhesion.
Objective To investigate the effects of carboxymethylchitosan- carboxymethylcellulose (CMCH-CMC) film on the adhesion and heal ing of colonic anastomosis. Methods Sixty-four healthy adult male SD rats was randomly divided into control group and experimental group (n=32). The model of colonic anastomosis was made according to Buckenmaier’ smethod in all rats. The experimental group was treated by wrapping anastomosis with CMCH-CMC film (3 cm × 2 cm) and the control group was not treated. At 7 days and 14 days after operation, the adhesion formation of colonic anastomosis was observed, the tensile strength of the anstomosis was assessed and compared with 6 normal rats, and the hydroxyprol ine (HP) content of the anastomotsis was detected. Results There were 3 deaths in the experimental group and 2 deaths in the control group. The adhesive scores of the experimental group on the 7th and 14th postoperative day [(0.50 ± 0.16) points and (0.45 ± 0.14) points, (Plt; 0.05)] were significantly lower than those of the control group [(1.67 ± 0.15) points and (2.29 ± 0.18) points, (P lt; 0.05)], (Plt; 0.01). Tensile strength were more marked on the 14th postoperative day than on the 7th postoperative day in the control group (Plt; 0.05), but there was no significant difference between the 7th day and the 14th day in the experimental group. The tensile strength of thecontrol group and the experimental group on the 14th postoperative day [(178.36 ± 20.10) and (172.74 ± 22.18) mmHg] were respectively higher than those on the 7th postoperative day [(138.67 ± 16.65) and (130.81 ± 18.38) mmHg] (Plt; 0.01). The tensile strength of the control group and the experimental group on the 7th postoperative day were respectively significantly lower than that of the normal rats (P lt; 0.01). The level of HP in the anastomosis was significantly higher on the 7th postoperative day in the experimental group [(84.47 ± 11.87) μg/mg dried weight] than that of the control group [(55.47 ± 12.89) μg/mg dried weight), (Plt; 0.05)], but there was no significant difference between the experimental group and the control group on the 14th postoperative day [(146.07 ± 14.81) μg/mg dried weight, (137.14 ± 16.81) μg/mg dried weight, (P gt; 0.05)]. Conclusion The CMCH-CMC film can decrease adhesion the formation of colonic anastomosis, but does not interfere with the heal ing of colonic anastomosis.
ObjectiveTo observe the effect of interleukin-8 (IL-8) on the adhesion and migration of retinal vascular endothelial cells (RCEC). MethodsA cell experiment. Human RCEC (hRCEC) was divided into normal control group (N group), advanced glycation end product (AGE) treatment group (AGE group), and AGE-induced combined IL-8 antagonist SB225002 treatment group (AGE+SB group). The effect of AGE on IL-8 expression in hRCEC was observed by Western blot. The effect of SB225002 on hRCEC migration was observed by cell scratch assay. The effects of SB225002 on leukocyte adhesion and reactive oxygen species (ROS) on hRCEC were detected by flow cytometry. Student-t test was performed between the two groups. One-way analysis of variance was performed among the three groups. ResultsCompared with group N, the expression level of IL-8 in cells of AGE group was significantly increased, with statistical significance (t=25.661, P＜0.001). Compared with N group and AGE+SB group, cell mobility in AGE group was significantly increased (F=29.776), leukocyte adhesion number was significantly increased (F=38.159, 38.556), ROS expression level was significantly increased (F=22.336), and the differences were statistically significant (P＜0.05). ConclusionIL-8 antagonist SB225002 may down-regulate hRCEC adhesion and migration by inhibiting ROS expression.
Objective To explore the effects of exogenous basic fibroblast growth factor (bFGF) on insheathed tendon healing and adhesion formation. Methods Ninety Leghorn chickens were randomly divided into 3 groups (groups A, B and C), 30 animals for each group, and the right third digitorum longus tendon of the chicken was transected to make defect models. In group A, the tendon was sutured in situ after transection. In group B, the tendon was sutured after 0.6 μl fibrin sealant (FS) was applied at repair site. In group C, the tendon was sutured after 0.6 μl FS mixed with 500 ng bFGF was appliedat repair site. At 1, 2, 4 and 8 weeks after operation, the tendons of 6 chickens in each group were harvested for morphological and histological evaluation. Six specimens of each group was obtained for biomechanical test at 8 weeks. Results The gross observation showed that the differences of grading of tendon adhesion were not significant between groups A, B, and C 8 weeks after operation(Pgt;0.05). Histological evaluation showedthat there were no significant differences in fibroblast counting and the content of collagen fibers between groups A and B(P＞0.05). The angiogenesis, fibroblast proliferation and collagen production in the sheath, epitendon and parenchyma at repair site in group C occurred earlier and were more than those in groups A and B, showing significant differences (Plt;0.05). The biomechanical tests showed that the gliding excursionof the tendon in group A, B and C were 3.44±0.43、3.51±0.56 and 2.84±0.42 mm respectively; the work of flexion were 14.87±1.72、14.08±1.85 and 20.62±3.52 Nmm respectively; the ultimate tensile strength of the tendon was10.34±1.45，11.26±1.83 and 15.02±2.20 N respectively; showing no significant differences between groups A and B(Pgt;0.05), but showing significant differences between group C and groups A, B(Plt;0.05). Conclusion The exogenous bFGF at tendon repair site can facilitate insheathed tendon healing, but also increase the tendon adhesion formation.
ObjectiveTo explore the relationship among plasma cytokines’ level, adhesion molecules expression and skin damage in patients with chronic venous insufficiency (CVI) of lower extremities.MethodsIn 32 patients with CVI and 8 normal individuals as control, blood TNFα, IL1β and IL2R were assayed with ELISA method; serum endothelial cellintercellular adhesion molecule1(ECICAM1), polymorphonuclearCD18(PMNCD18) and polymorphonuclearCD11b(PMNCD11b) were assayed with immunohistochemical method; and ultrastructure of diseased veins was examined by electroscope.ResultsThe results showed that the level of plasma TNFα and IL1β increased remarkably in Class 2-3 compared with Class 1 and control (P＜0.05), IL2R had no difference in Class 1,2,3(Pgt;0.05). The index of ECICAM1 and PMNCD11b positively expression increased remarkably in Class 2-3 compared with that in Class 1 and control. The index of PMNCD18 expression in Class 2-3 and Class 1 was greatly higher than that in control (P＜0.05). The expression of ICAM1 was positively correlated with that of CD11b/CD18. Electron microcopy showed that the change in microvessel was mainly PMN adhesion with endothelial cells (ECs) and trapped in microvessels.ConclusionThe results suggest that activated monocyte may release TNFα and IL1β, upregulate ICAM1 and CD11b/CD18 expression, and mediate the PMN adhesion to ECs, thus causing ECs and tissue damage. It may be one of important mechanism of venous ulcer.
ObjectiveTo identify the expression functions of human NF-κBp65 nuclear localization signals' deletion mutant plasmids(namely pcDNA3.1(+)-NF-κBp65ΔNLS, NF-κBp65ΔNLS, for short) and the changes of proliferation, migration and adhesion ability of A549 lung cancer cells with low expression of NF-κBp65 (namely A549/NF-κBp65 shRNA cells). MethodsHuman A549/NF-κBp65 shRNA cells were cultivated and divided into a control group, a transfection pcDNA3.1 (+) group, and a transfection NF-κBp65ΔNLS group. Indirect immunofluorescence, real-time fluorescent quantitative PCR and Western blot techniques were used to detect the NF-κBp65 intracellular localization and the change of NF-κBp65 mRNA and protein expression level. MTT, Transwell and cell adhesion experiments were used to analyze the changes of proliferation, migration and adhesion ability of A549/NF-κBp65 shRNA cells. ResultsThe human NF-κBp65ΔNLS eukaryotic expression plasmid was successfully constructed. Compared with the control group and the transfection pcDNA3.1(+) group, NF-κBp65 mRNA expression level in A549/NF-κBp65 shRNA cells was increased in the transfection NF-κBp65ΔNLS group(10.63±0.84 vs. 1.04±0.21 and 1.23±0.22, P < 0.01) and NF-κBp65 protein expression level was also increased (1.07±0.06 vs. 0.53±0.02 and 0.59±0.04, P < 0.01). NF-κBp65 protein mainly located in the cytoplasm, and did not significantly transferred into the nucleus after stimulated by TNF-α. At the same time, A549/NF-κBp65 shRNA cells' proliferation, migration and adhesion ability were enhanced compared with the control group and the transfection pcDNA3.1(+) group. ConclusionsThrough gene mutation technology to build the human NF-κBp65ΔNLS eukaryotic expression plasmid and transfect into A549/NF-κBp65 shRNA lung cancer cell lines, both mRNA and protein expression levels of NF-κBp65 were increased significantly, and NF-κBp65 protein mainly located in the cytoplasm. The overexpressed NF-κBp65 in cytoplasm can obviously enhance the A549/NF-κBp65 shRNA cell's proliferation, migration and adhesion ability. It suggests that NF-κBp65 stranded in the cytoplasm can still regulate biological behavior of lung cancer cells by influencing the NF-κB signaling pathway related proteins.