Some Chinese traditional medicines were found to inhibit rejection of graft. The antirejection effects of chuanxiong, LCH and HXI in thyroid allografts of rabbits were studied for selecting an immune depressor from Chinese traditional medicine with efficient and less sideeffect. The rabbits were divided into 5 groups in the study, with 7 in each group. Group I: The control group, no drug was used. Group II: dexamethason 0.25mg/kg/day, intramuscularly. Group III: chuanxiong water solution, 5g/kg/day, orally. Group Ⅳ： LCH water solution, 10g/kg/day, orally. Group Ⅴ： HXI water solution, 6g/kg/day, orally. The medication was given for 28 days. The grafted thyroids were removed for histopathological examination on the 28th day postoperatively and were scored and classified. The rejection and the survival of grafts were scored and classfied according to the La Rosa and Warrens criterion. The histopathological findings were as following: in Group I, follicles were badly damaged with much lymphocytes infiltration and fibrosis; in Gracup Ⅱ， two rabbits died, the other three showed damaged of the thyroid tissue and much lymphocytes infiltration; in group Ⅲ and Ⅴ, three cases showed damage of thyroid tissue, however, better revascularization was evident in Group Ⅲ; in Group Ⅳ, there was one case with much lymphocytes infiltration. It seemed that the degree of damage of grafts in the experimental groups was better than that in the control group, and had less lymphocytes infiltration, especially in Group Ⅳ. It was suggested that chuanxiong, LCH, HXI and dexamethason could protect the grafted thyroid, but the sideeffect of dexamethason was more than the other three. The antirejection of LCH was the best of the three. It was worth doing more research. HXI.
Objective To provide a reliable experimental model for gastroesophageal reflux (GER) study. Methods Twenty Japan 5-month-old male rabbits wererandomly divided into two groups: group cardiomyotomy(n=10), group partial cardiomyectomy(n=10). The operations of cardiomyotomy and parital cardiomyectomy were performed in 2 groups respectively. All the animals underwent intraesophagealpH detection 1 week before operation and 4 weeks after operation. The mean changes of reflux ratios were compared between before operation and after operation.Results In gastroesophageal reflux ratio between before operation and after operation, there was no significant difference in group cardiomyotomy (1.98%±1.52% and 4.32%±2.39%, Pgt;0.05) and there was significant difference in group partialcardiomyectomy（1.56%±1.57% and 13.56%±3.27%, Plt;0.05）. Conclusion The reliable experimental model of GER can be made with procedure of partial cardiomyectomy. It can be used in estimating the operative procedure of antireflux and is conducive to dynamic observation and study of esophagitis.
To investigate the effect of propofol intra-aortic and intravenous infusion on the concentration of propofol for an ischemia-reperfusion spinal cord injury in rabbits. Methods Forty-six healthy adult New Zealand white rabbits were randomly divided into 3 groups: sal ine infusion group (group N, n=10), propofol intra-aortic infusion group (group A, n=16) and propofol intravenous infusion group (group V, n=16). The infrarenal abdominal aorta was occluded for 30 min during which propofol 50 mg/kg was infused continuously intra-aortic or intravenous with a pump in group A and V. In group N, the same volume of normal sal ine was infused in the same way and at the same rate as in group A. Upon reperfusion, propofol concentration of the spinal segments of L4-6 and T6-8 was examined in group A and V. At 48 hoursafter reperfusion, the neurological outcomes were recorded in each group. Results Mean blood pressure in group V from the time of 5 minutes after occlusion decreased more than in group N (P lt; 0.05) and than in group A from the time of 10 minutes after occlusion(P lt; 0.05). The mean blood pressure in group N increased more than in group A from 15 minutes after occlusion (P lt; 0.05). The heart rate increased more in group V from 10 minutes after occlusion than in group N and A (P lt; 0.05) in which no difference was observed. The propofol concentration in L4-6 of group A (26 950.5 ± 30 242.3) ng/g was higher than that in T6-8 of group A (3 587.4 ± 2 479.3) ng/g and both L4-6 (3 045.9 ± 2 252.9) ng/g and T6-8 (3 181.1 ± 1 720.9) ng/g of group V(P lt; 0.05). The paraplegia incidence was lower (30%) and the median of normal neurons was higher (8.4) in group A than in group N (80%, 2.2) and group V(100%, 1.9), (P lt; 0.05). There was no significant difference in group N and V in paraplegia incidenceand the median of normal neurons (P gt; 0.05). Conclusion Intra-aortic infusion shows a better neurological outcome than intravenous infusion and could contribute to higher concentration of propofol in the ischemia spinal cord.
【摘要】 目的 研究双侧迷走神经切断对肺缺血再灌注引起的氧化应激反应的影响。 方法 将24只健康雄性新西兰大白兔随机分为:假手术组（S组）、缺血再灌注组（IR组）、双侧迷走神经切断合并缺血再灌注组（NIR组）。缺血前和再灌注末抽取动脉血进行血气分析，观察动脉血氧分压PaO2及肺泡动脉氧分压差（A-aDO2）的变化。再灌注末取肺组织检测肺的湿干重比值（W/D）和氧化应激指标，包括丙二醛（MDA）、超氧化物歧化酶（SOD）及过氧化氢酶（CAT）。 结果 与S组比较，缺血再灌注明显降低了PaO2，增加了A-aDO2和W/D值，增加了肺组织MDA含量并降低了SOD、CAT活性；双侧迷走神经切断进一步降低了SOD活性。 结论 切断实验兔的双侧迷走神经，降低了肺组织抗氧化酶-超氧化物歧化酶的活性，提示迷走神经在降低肺缺血再灌注引起的氧化应激反应中发挥了重要的调节作用。【Abstract】 Objective To evaluate the effect of bilateral vagal nerves transection on lung ischemia-reperfusion induced oxidative stress. Methods A total of 24 New Zealand male rabbits were randomly divided into 3 groups: sham group (S group), ischemia-reperfusion group (IR group), and bilateral vagal nerves transection with ischemia-reperfusion group (NIR group). Before ischemia and at the end of reperfusion, arterial blood samples were collected for blood gas analysis. Arterial partial pressure of oxygen (PaO2) and alveolo-arterial oxygen tension difference (A-aDO2) were detected. At the end of reperfusion, lung tissues were obtained to measure wet/dry weight ratio (W/D). Evaluation of oxidative stress indicators, including content of lung malondialdehyde (MDA), superoxide dismutase enzyme (SOD) and catalase (CAT) activities was also performed. Results Compared with the S group, lung ischemia-reperfusion significantly decreased the PaO2, elevated A-aDO2 and lung W/D weight ratio. At the same time, MDA level in the lung tissue was elevated and SOD and CAT activities were decreased. After bilateral vagal nerves transection, SOD activity was further decreased. Conclusion Transection of bilateral vagal nerves reduced the activity of antioxidant enzyme, especially superoxide dismutase in lung tissue, suggesting that the integrity of the vagal nerves plays an important regulatory role in ischemia-reperfusion mediated oxidative stress in the lung.
ObjectiveTo investigate the effect of zinc finger protein A20 on lumbar intervertebral disc degeneration in rabbits.MethodsTwenty-six 3-month-old New Zealand rabbits, 2.0-2.5 kg in weight, were used to establish the model of intervertebral disc degeneration at L3, 4, L4, 5, and L5, 6 by transabdominal needle puncture. At 4 weeks after operation, the 24 rabbits were randomly divided into 4 groups after successful modeling, which checked by MRI. The target intervertebral discs of each group were injected with zinc finger protein A20 overexpressed adenovirus (Ov-A20 group), empty carrier adenovirus (NC group), phosphate buffer saline (control group), and shRNA-A20 adenovirus (Sh-A20 group). The biological responses of animals in each group were comprehensive scored before 1 day of injection and after 1, 2, 3, and 6 days of injection. At 2, 4, and 8 weeks after injection, the animals in each group were observed by MRI to obtain the exact T2 relaxation time (T2 signal value). After MRI examination, the animals were killed to take the degenerative intervertebral disc tissue; and the tissue was detected by Alcian blue staining to observed the intervertebral disc degeneration. The expressions of zinc finger protein A20, collagen Ⅱ, and aggrecan were detected by immunohistochemistry staining. The expressions of zinc finger protein A20, nuclear factor κB binding protein [P65, phosphate P65 (P-P65), collagen Ⅱ, aggrecan], inflammatory factors [tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β)], autophagy-related protein [LC3 (LC3Ⅱ/LC3Ⅰ) and P62] were detected by Western blot.ResultsThe comprehensive score of biological response in each group after injection was significantly lower than that before injection (P<0.05). At 6 days after injection, the comprehensive score of biological response in the Sh-A20 group was significantly lower than that in other groups (P<0.05), and there was no significant difference among other groups (P>0.05). The detection of MRI showed that the T2 signal value in the Ov-A20 group was the highest at 2, 4, and 8 weeks after injection (P<0.05), and the T2 signal value in the Sh-A20 group was the lowest at 2 and 4 weeks after injection (P<0.05). There was no significant difference between other groups (P>0.05). Alcian blue staining showed that the expression of aggrecan was the highest in Ov-A20 group and the lowest in Sh-A20 group at 4 weeks (P<0.05); the expression of aggrecan in Ov-A20 group was the highest at 8 weeks (P<0.05), and there was no significant difference between other groups (P>0.05). Immunohistochemical staining showed that the expressions of zinc finger protein A20, collagen Ⅱ, and aggrecan were the highest in Ov-A20 group and lowest in Sh-A20 group (P<0.05). Western blot showed that the expressions of zinc finger protein A20, collagen Ⅱ, aggrecan, and LC3 (LC3Ⅱ/LC3Ⅰ) proteins were the highest in the Ov-A20 group and the lowest in Sh-A20 group (P<0.05), while the expressions of P-P65, TNF-α, IL-1β, and P62 proteins were the lowest in Ov-A20 group and the highest in Sh-A20 group (P<0.05). There was no significant difference in the expression of p65 protein between groups (P>0.05).ConclusionZinc finger protein A20 can effectively regulate the process of lumbar intervertebral disc degeneration in rabbits by inhibiting inflammation.
ObjectiveTo study the long-term prevention effect of self-developed chitosan electrospun membrane on cerebrospinal fluid leakage. MethodsTwenty-five healthy adult New Zealand rabbits were selected to prepare the bilateral dural defect (0.8 cm×0.8 cm in size) via midline incision of head.Defect of the right was repaired with chitosan electrospun membrane as the experimental group; defect of the left was not repaired as the control group.At 2-16 weeks after operation,one rabbit was sacrificed for the general observation of inflammatory response surrounding bone window and absorption of chitosan electrospun membrane; at 3 and 6 weeks after operation,5 rabbits were sacrificed for sampling to observe histological change and collagen expression by HE and Masson staining,and to measure the expressions of epidermal growth factor receptor (EGFR) and basic fibroblast growth factor (bFGF) by immunohistochemical staining. ResultsNo inflammatory reaction of swelling,exudation,and sppuration appeared in the skin and subcutaneous tissue after operation in 2 groups.There was no adhesion around the chitosan electrospun membrane,and new fiber membrane formed under the chitosan electrospun membrane in the experimental group; no cerebrospinal fluid leakage happened; the chitosan electrospun membrane was gradually degraded with time,and was completely absorbed at 16 weeks.There was uneven scar around the dural detect in control group.Histological observation showed less inflammatory cell infiltration in the experimental group,showing significant difference in the number of inflammatory cells compared with control group at 3,6 weeks (P<0.05); capillary,granulation tissue and collagen fiber massively proliferated; collagen fiber arranged in line,and there was a clear borderline between chitosan electrospun membrane and adjacent collagen fiber.The immunohistochemical staining showed that there were high expressions of bFGF and EGFR in the experimental group,and low expressions of bFGF and EGFR in the control group. ConclusionChitosan electrospun membrane for dural defect of rabbit can effectively reconstruct the dura,and it has exact long-term prevention effect on cerebrospinal fluid leakage.
Objective To evaluate the effect of WO-1 on repair of the bone defect in the New Zealand rabbit radius by an oral or local administration. Methods Bone defects were surgically created in the bilateral radii of 36 Zealand rabbits (1.6-2.0 kg), which were randomly divided into3 groups. In Group A, the defective areas were given WO-1 0.1 ml (50 mg/ml) by the local injections; in Group B, the rabbits were given WO-1 5 mg each day by the oral administration. Group C was used as a control group. Among each of the 3 groups, 4 rabbits were randomly selected and were sacrificed at 20, 30 and 60 days after operation, respectively. Then, the serological, X-ray and histological examinations were performed. Results The serum alkaline phosphatase and bone glaprotein levels were significantly higher at 20 and 30 days after operation in Groups A and B than in Group C, but significantly lower at 60 days after operation in Groups A and B than in Group C（Plt;0.01）. The X-ray and histological examinations at 20, 30 and 60 days after operation revealed that the callus formation and remodeling were earlier in Groups A and B thanin Group C, and the remodeling was earlier and better in Group A than in Group B. Conclusion WO-1 can promote the repair of the radial defect in a rabbit; however, further studies on the doseeffect relationship, administration time, and administration route are still needed.
Objective Bioactive borate glass (BG) has good biocompatibil ity and biodegradation. To investigate the feasibilty of bioactive borate glass as a carrier of the antibiotic controlled-releasing by implanting vancomycin-loaded BG (VBG)into the focus of tibia chronic osteomyel itis after debridement. Methods VBG and vancomycin-loaded calcium sulfate (VCS) were prepared with a vancomycin content of 80 mg/g. Sixty-five New Zealand white rabbits, weighing 2.12-3.91 kg (mean, 2.65 kg), were used. The tibia chronic osteomyel itis rabbit models were establ ished by injecting methicill in-resistant Staphylococcus aureus (MRSA, 0.1 mL, 1 × 109 cfu/mL) into the right tibia of 65 rabbits. After 3 weeks of injection, 54 rabbits of successful models were randomly divided into groups A (n=11), B (n=11), C (n=16), and D (n=16). Simple debridement was performed in group A; BG, VCS, and VBG were implanted into the infection sites of groups B, C, and D respectively after thorough debridement. A sample of the debrided tissues was harvested for bacterial examination. The vancomycin serum levels were determined in groups C and D at 1, 2, 4, 10, 24, and 48 hours after operation. The boron serum levels were determined in groups B and D at 10, 24, 48, 72, and 120 hours after operation. After 8 weeks, the effectiveness was assessed radiographically, bacteriologically, and histopathol ogically. Results Ten rabbits died after operation. No vancomycin was detected in group C; the vancomycin level increased gradually, reached the highest level at 4 hours after operation, and then decreased rapidly in group D. No boron was detected in group B; the boron reached the highest serum level at 10 hours after operation, and then decreased gradually in group D. At 8 weeks, calcium sulfate degraded in group C; BG degraded partially in group D; and no obvious degradation was observedin group B. The repair effect was better in group D than in group C. There was no significant difference in radiograph scoring between groups A, B, C and D (P gt; 0.05) before operation, but there was significant difference between group D and groups A, B, C (P lt; 0.05) at 8 weeks after operation. The bacterial culture showed that all the MRSA results were positive in 4 groups. At 8 weeks, the negative rates of MRSA examination were 36.36%, 18.18%, 73.33%, and 81.25% respectively in groups A, B, C, and D, showing significant differences between group D and groups A, B (P lt; 0.05). The histopathological observation showed that a large number of new bones formed and no foreign body reaction occurred in group D. The histopathologic scores of groups A, B, C, and D were 6.45 ± 3.62, 7.55 ± 3.36, 4.27 ± 2.91, and 3.81 ± 3.04 respectively, showing significant differences between group D and groups A, B, and between group C and group B (P lt; 0.05). Conclusion VBG can improve the repair of bone defect in the treatment of chronic osteomyel itis.
Objective To repair the defects in articular cartilage with collagen complex gradient TCP in vivo andto study the regenerated cartilage histomorphologically. Methods The models of defects in articular cartilage were madeartificially in both condylus lateral is femoris of mature rabbits, male or female, with the weight of 2.0-2.5 kg. The right defects were implanted with the material of Col/TCP as the experimental group and the left defects were untreated as the control group. The rabbits were killed at 4, 6, 8, 12 and 24 weeks after operation, respectively, with 6 ones at each time, and the macroscopic, histological, ultrastructural examinations and semi-quantity cartilage scoring employing Wakitanifa repaired cartilage value system were performed. Results Four weeks after operation, the defects in the experimental group were partly filled with hyal ine cartilage. Twelve weeks after operation, the defects in the experimental group were completely filled with mature hyal ine cartilage. Twenty-four weeks after operation, regenerated cartilage had no ataplasia. However, fibrous tissues were seen in the control group all the time. At 4, 6, 8, 12 and 24 weeks ostoperatively, the Wakitanifa cartilage scores were 7.60 ± 0.98, 5.69 ± 0.58, 4.46 ± 0.85, 4.35 ± 0.12 and 4.41 ± 0.58, respectively, in the experimental group and 10.25 ± 1.05, 9.04 ± 0.96, 8.96 ± 0.88, 8.88 ± 0.68 and 8.66 ± 0.54, respectively, in the control group. At 4, 6, 8, 12 and 24 weeks postoperatively, the collagen II contents were 0.28% ± 0.01%, 0.59% ± 0.03%, 0.68% ± 0.02%, 0.89% ± 0.02% and 0.90% ± 0.01%, respectively, in the experimental group, while 0.08% ± 0.02%, 0.09% ± 0.04%, 0.11% ± 0.03%, 0.25% ± 0.03% and 0.29% ± 0.01%, respectively, in the control group. Differences between the control group and the experimental group were significant (P lt; 0.05). By then, typical chondrocyte was observed by transmission electron microscope in the experimental group and much fiber with less fibrocyte was observed in the control group. Conclusion Three-dimensional scaffold collagen complex gradient TCP may induce cartilage regeneration to repair the defects of articular cartilage in vivo.
Objective To research the effect of porcine acellular dermal matrix in the reconstruction of abdominal wall defects in rabbits, and to investigate the appl ication feasibil ity of xeno-transplantation of acellular dermal matrix. Methods The porcine acellular dermal matrix was prepared from a health white pig. Twenty-six Japanese white rabbits (weighing 2.2-2.3 kg, female or male) were randomly assigned to 2 groups: the control group (n=6) and the experimental group (n=20). In the control group, the full-thickness abdominal wall defect of 5.0 cm × 0.5 cm was made, and the defect wassutured directly; in the experimental group, the full-thickness abdominal wall defect of 5.0 cm × 2.5 cm was made, and the defect was repaired with porcine acellular dermal matrix patch at the same size as the defect. At 5 weeks after surgery, the incidence of hernia and the intra-abdominal adhesions were observed and the wound breaking strength was compared between the patchfascia interface and the fascia-fascia interface. The graft vascularization was evaluated through histological analysis at 6 months after surgery in the experimental group. Results No hernia occurred in all rabbits of 2 groups. At 5 weeks after surgery, heal ing was observed between patch and the muscularfascia; the vascularization was seen in the porcine acellular dermal matrix patch. There was no significant difference in the adhesion grade (Z= —0.798, P=0.425) between the experimental group (grade 2 in 1 rabbit, grade 1 in 5, and grade 0 in 12) and the control group (grade 1 in 1 and grade 0 in 5). No significant difference was found (t= —0.410, P=0.683) in the breaking strength between the patch-fascia interface in the experimental group [(13.0 ± 5.5) N] and the fascia-fascia interface in control group [(13.6 ± 4.0) N]. In the experimental group, the small vessels and the infiltration of inflammatory cells were observed in the porcine acellular dermal matrix patch after 5 weeks through histological observations. The junctions of the patch-fascia interface healed with fibrous connective tissue. At 6 months after surgery, the inflammation was subsided and the collagen fiber of the patch was reconstructed. Conclusion The porcine acellular dermal matrix patchhas good results in repairing full-thickness abdominal wall defect. The patch-fascia interface has siml iar breaking strength to the fascia-fascia interface. The collagen fibers of the patch are reconstructed.