Objective To evaluate the effect on microvessel density (MVD) and vascular endothelial growth factor (VEGF) expression of combining radiofrequency ablation (RFA) with arsenious acid (AA) locally treating liver VX2 tumor in rabbits. Methods Twenty-eight New Zealand White rabbits with implanted liver VX2 tumors were randomly divided into four groups, control group (n=7), AA group (n=7), RFA group (n=7) and combination (RFA+AA) group (n=7). All rabbits were killed 14 days after treatment. MVD and VEGF expression were examined by immunohistochemistry. Results The MVD degraded one by one in control group,AA group,RFA group and RAF+AA group, which were (38.50±0.44), (23.07±0.47), (18.65±0.39) and (11.36±0.36)/HP respectively, compared while each two groups, P＜0.05. The VEGF expression also degraded one by one, the ratio of positive cases were 7/7, 5/7, 4/7 and 2/7 respectively, compared while each two groups, P＜0.05. There was positive correlation between VEGF expression and MVD (Person conefficient of product-moment correlation r=0.47, P＜0.01). Conclusion Combining RAF with AA therapy can greatly decrease MVD and VEGF expression of tumor tissue.
Objective To explore the best flexion angle of the transplantation tendon for fixing joint in simultaneously reconstructing of the anterior cruciate l igament (ACL) and posterior cruciate l igament (PCL) using semitendinosus tendon as autologous graft. Methods Twenty-four clean level New Zealand White rabbits [(aged 6-8 months, male or female, and weighing (2.5 ± 0.2) kg] were selected and divided randomly into 3 groups (n=8) according to fixation angle of the reconstructed l igaments. The bilateral semitendinosus tendons of hind legs were used to reconstruct the PCL and ACL of right hind leg, and the reconstructed l igaments were fixed at knee flexion angles of 90° (group A), 60° (group B), and 30° (group A). The rabbit general situation was observed after operation, and the specimens of the knee joints (including 10 cmdistal end and 10 cm proximal end) were harvested for testing extension and flexion, displacement, and internal and external rotation at 3 months after operation. Results All the rabbits survived to the end of experiment. There was no significant difference in maximal displacements of ACL and PCL among 3 groups (P gt; 0.05). The anterior and posterior displacements of shift in 3 groups were less than 1 mm, suggesting good stabil ity. The anterior displacement and the posterior displacement at 30° flexion and 90° flexion in group A were significantly larger than those in group C (P lt; 0.05). There were significant differences in internal rotation angle and external rotation angle between group A and group C (P lt; 0.05), and there was no significant difference among other groups (P gt; 0.05). Conclusion When simultaneously reconstructing ACL and PCL, the knee flexion angle of 60° for fixing the reconstructed l igaments can achieve the best effect.
Objective To study the effect of two cytokines, basic fibroblast growth factor(bFGF) and insulin-like growth factor-I(IGF-I), on cell proliferation in chondrocytes of adult rabbits. Methods The primary chondrocytes of adult rabbits were harvested and cultured with bFGF and IGF-I at different concentrations,respectively, as well as with the mixture of the two cytokines; the quantity of cultured chondrocytes was detected by MTT assay at the 24th, 48th and 72th hours; and the final fold increase of different groups was measured by cell count for the 3rd passage; and the proliferation index of the groups was recorded by flowing cytometer on the 14th day. Results ① The cultured chondrocytes with either bFGF, IGF-I or their mixture were significantly more than that of control group at the 24th, 48th and 72th hours (P＜0.01). ② After the 3rd passage, the final folds of proliferation were significantly higher in the groups with cytokinesthan in the control group (P＜0.01); and the final fold with the mixture ofcytokines was significantly higher than that of both IGF-I and bFGF (P＜0.01). ③ Theproliferation index was significantly higher in the groups with cytokines than in the control group (P＜0.01); the proliferation index with the mixture of cytokines was significantly higher than that of both IGF-I and bFGF (P＜0.05); besides, proliferation index was higher when cytokine was applied twice than once (P＜0.05). Conclusion bFGF and IGF-I could promote chondrocytes proliferation of adult rabbits obviously and they are synergistic in cell proliferation.
In order to repair cartilage defect in joint with transplantation of cryopreserved homologous embryonic periosteum, 30 rabbits were used and divided into two groups. A 4 mm x 7 mm whole thickness cartilage defect was made in the patellar groove of femur of each rabbit. The homologous embryonic rabbit skull periosteum (ERSP), preserved in two-step freezing schedule, was transplanted onto the cartilage defect of joints of one group and autogenous periosteal graft was done in the joint defect of the other group. The knees were not immobilized, following operation and 16 weeks later, the newly formed tissue in the defects were assessed by gross observation, histochemical examination and biochemical analysis. The results showed that new hyaline-like cartilage was formed in the cryopreserved ERSP grafted knee, and had no significant difference from that of the knee receiving autogenous periosteal graft, but had significant difference from that of the fresh ERSP grafted knee and the non-grafted knee. Furthermore, the new hyaline-like cartilage had the biochemical characteristics of a fibrous cartilage. The conclusion was that this method might be feasible to repair articular cartilage defects.
We compared the sensitivities of human embryo conjtmctival fihroblasts(HECF)and rabbit conjunctival fibroblasts(RCF)m five anlineoplasties in vitro.When the concentration of vincrismum and doxorubicin was 0.001~10mg/L.5-FU was 1~1000mg/L and cisplatin was 0.01~10mg/L,the sensitivities of HECF to tile drugs were lower than that of RCF (Plt;0.01).while the difference of the sensitivilics be;ween HECF and RCF to VP-16 was not significant (P＜0.05). The results suggested that the selection of therapeutic agents for intraocular proliferative disease wilh'hunmn conjunctiva fihroblasts may be more valuable than that with RCF. （Chin J Ocul Fundus Dis,1994,10:223-225）
An experimental model of proliferative vitretinopathy(PVR) induced by macrophages was used for the evaluation of drug efficacy of daunomycin encapsulated in liposomes in the treatment of PVR.Five mu;g daunomycin(n=40),10mu;g daunomycin-liposome(DL,n=30)and 0.1 ml saline or empty liposomes(n=40,as controls)were injected into the rabbit vitreous after macrophage injection.Retinal detachment developed in 77.5% of the control eyes on day 28,compared to 33.3% of the eyes treated with DL(P＜0.01)and 50% of the daunomycin-treated eyes(P＜0.05).The results suggest that encapsulation in liposomes of cytotoxic agents can enhance drug efficacy.The phasic course of development of PVR is important in the selection of particular drugs. （Chin J Ocul Fundus Dis,1993,9:77-80）
Objective To investigate the influence of colectomy on the expressions of 5-hydroxy tryptamine (5-HT) and chromogranin A (CgA) in colon mucosa of Chinchilla rabbits. Methods Colon (7-8 cm) upon colon-rectum junction (control group) of 15 Chinchilla rabbits was cut out. After two weeks, these rabbits were executed and the samples of colon at anastomotic stoma (study group) were taken. 5-HT positive cells and CgA positive cells in two groups were detected by immunohistochemical method. Results The number of 5-HT positive cells was 10.40±2.22 in control group， and 26.27±2.35 in study group; the number of CgA positive cells was 20.60±5.34 in control group, and 51.51±6.13 in study group. There were significant differences between two groups respectively (P＜0.01). Conclusion The increase of 5-HT positive cells and CgA positive cells can be caused by colectomy.
This research was aimed to study the effect of Emodin gel on the hypertrophic scars of rabbit ears. A total of 18 rabbits were randomly divided into Emodin group (9 rabbits) and control group (9 rabbits) after the successful animal model for hypertrophic scars had been made. The rabbits in the Emodin group were treated with Emodin Gel,while no special treatment was given to those in the control group. The other living conditions were all kept the same in the two groups. The diameter,hardness, and expression of transforming growth factor-β (TGF-β) and interleukin-1 (IL-1) of hypertrophic scars were measured after 4 weeks. Transmission electron microscopy was applied to observe the ultra-structure of the fibroblasts of hypertrophic scars. But there was no difference between the two groups in the diameter of hypertrophic scars (P>0.05). The hardness, expression of TGF-β and IL-1 in hypertrophic scars in the Emodin group decreased, compared to the control group (P<0.05, P<0.01, P<0.05). Transmission electron microscopy showed that the fibroblast and organelle lessened in the cytoplasm and the collagen fibers dissolved obviously. The study showed that Emodin gel decreased the hardness of hypertrophic scars in the rabbit ears, and inhibited the proliferation of fibroblasts in local area. Therefore, Emodin gel treatment would be one of the methods to prevent and treat hypertrophic scars.
Objective To explore the protective effect and mechanism of Astragalus polysaccharides (APS) on liver injury in the state of brain death in New Zealand rabbits. Methods Twenty-four New Zealand rabbits were randomly divided into 3 groups (n=8): the blank control group, the brain death group, and the APS group. We obtained blood and liver tissue specimens from rabbits of three groups at 4 h and 8 h after treatment respectively (n=4). The rabbits of blank control group simulated the procedures of anesthesia and surgery of the brain death, without the Foley balloon catheter being pressurized, and maintained anesthesia. The brain death group: brain-dead models were established. The APS group: injection of APS (12 mg/kg) via the femoral vein bolus immediately after anesthesia, brain-dead models were established as same as rabbits of brain death group. The blood and liver tissue samples were taken at 4 h and 8 h after treatment to detect aminotrans-ferase (AST), alanine amino-transferase (ALT) and tumor necrosis factor α (TNF-α), and to observe the change of liver tissue by HE staining and immunohistochemical staining〔expression level of nuclear transcription factor p65 protein (NF-κB p65) could be detected by immunohistochemical staining〕. Results ① ALT and AST. Compare with the blank control group at the same time (4 h and 8 h), levels of ALT and AST in brain death group and APS group were significantly increased (P<0.05), and the levels of ALT and AST in brain death group were higher than those of APS group at each time point (P<0.05). In the same group, compared with 4 h, there was no significant difference in the levels of ALT and AST in blank control group at 8 h (P>0.05); the levels of ALT and AST in brain death group at 8 h were both higher than those of 4 h (P<0.05); the levels of ALT at 8 h in APS group was higher than that of 4 h, but there was no significant difference in the level of AST between 4 h and 8 h (P>0.05). ② TNF-α. Compare with the blank control groups at same time (4 h and 8 h), levels of TNF-α in brain death group and APS group were significantly increased(P<0.05), and level of TNF-α in brain death group was higher than that of APS group at 4 h and 8 h (P<0.05). ③ The HE results. The liver tissue structure of blank control group, brain death group, and APS group at 4 h had no obvious change. The liver tissue structure of brain death group at 8 h showed the evident tissue damage: liver cells showed the balloon samples, disordered arrangement, cytoplasmic loose light dye net-like, and inflammatory cells infiltrated in portal area. The liver tissue structure of APS group at 8 h showed that, liver cells showed mild edema, normal arrangement, and a small amount of inflammatory cells infiltrated in portal area. The liver tissue structure damage of APS group at 8 h was milder than that of brain death group. ④ Immunohistochemical staining results. There was no significant difference in expression levels of NF-κB p65 protein among blank control group, brain death group, and APS group at 4 h (P>0.05). But at 8 h, the expression levels of NF-κB p65 protein in brain death group and APS group were higher than that of blank control group (P<0.05), and the expression level of NF-κB p65 protein in brain death group was higher than that of APS group (P<0.05). The expression levels of NF-κB p65 protein in brain death group and APS group at 8 h was higher than that of 4 h in the same group (P<0.05), but there was no significant difference between 4 h and 8 h in blank control group (P>0.05). Conclusions Brain death will cause liver damage and the injury degree may be related to the continuous time. The damage at 8 h was more serious than that of 4 h. APS has a protective effect on liver of brain-dead rabbits' and its mechanism may be closely related to inhibit TNF-α and NF-κB by diverse ways to reduce the inflammation of the liver injury.
Objective To investigate the effect of ultrasonic irradiation time on enhanced green fluorescent protein (EGFP) gene transfection efficiency and local tissue in bone defects using ultrasound-mediated microbubble destruction. Methods Thirty 3-month-old New Zealand rabbits (2.5-3.0 kg in weight) were randomly divided into 5 groups (n=6) and bone defect models were made on the right ulna. At 10 days after modeling, suspension of microbubbles and EGFP plasmids were locally injected (0.3 mL/kg) and then ultrasound was performed on defect at a frequency of 1 MHz, a intensity of 0.5 W/cm2, and a duty ratio of 20% for 1, 2, 3, 4, and 5 minutes respectively (in 1, 2, 3, 4, and 5 minutes groups respectively). The survival condition was observed. Rabbits were sacrificed for gross observation at 7 days after transfer. The gene expression was observed by fluorescence staining. HE staining and transmission electron microscopy were used to observe the local tissue damage. Results The animals all survived. New soft tissue formed in bone defects area at 1 week after transfer, the surrounding muscle tissue was partly filled in it. Green fluorescence expression was observed in all rabbits. The expression was the strongest in 2 minutes group, and was the weakest in 1 minute group. The absorbance (A) value showed significant differences when compared 1 minute and 2 minutes groups with other groups (P<0.05), but no significant difference was found between 3, 4, and 5 minutes groups (P>0.05). Tissue damage was observed in all groups and it was aggravated with the increase of irradiation time. Conclusion EGFP transfection efficiency in bone defect by ultrasound-mediated microbubble destruction is related to irradiation time. EGFP gene can be efficiently transfected without obvious toxicity at 1 MHz, 0.5W/cm2, and duty ratio of 20% for 2 minutes in bone defects of rabbits.