目的 建立犬开放性气胸海水浸泡的实验模型 ,探讨实验动物早期死亡原因。 方法 2 0条健康成年杂种犬随机分为两组。对照组 :实验动物受伤后直接观察 ;实验组 :动物受伤后置入人工配制的海水中。监测血流动力学、呼吸、血液渗透压、血液电解质、动脉血气变化以及肺部病理改变。 结果 实验组死亡率明显高于对照组 ,平均生存时间为 45分钟。实验组经海水浸泡后有急性呼吸和循环功能衰竭、严重电解质平衡紊乱、高渗血症、重度肺损伤以及严重代谢性和呼吸性酸中毒。 结论 开放性气胸后海水浸泡可引起一系列严重的病理生理变化 ,其结果是导致实验动物早期死亡的重要原因。
Objective To investigate the result of the transplantation of frozen canine phalangeal joint allografts perforated and incorporated with autogenic bone marrow. Methods A proximal interphalangeal joint defect of 1.5 cm was prepared at bilateral sides of twenty-four adult healthy out-bred dogs. Three different types of allografts were applied to repair the defects: fresh autogenic phalangeal joints (group A,n=16), frozen phalangeal joint allografts perforated and incorporated with fresh autogenic bone marrow(group B, n=16), and frozen phalangeal joint allografts(group C, n=16). Radiographic and histological study wereused to evaluate the survival of transplanted joints. The observation was done 1, 3, 6 and 12 months after operation respectively. Results Based on the radiographic and histological changes of the transplanted joints, the osteoarthropathy of transplanted canine phalangeal joints could be divided into 3 degrees: mild degeneration, moderate degeneration and severe degeneration. Mild degeneration was observed in group A from 3 to 12 months. Mild degeneration was also found in group B from 1 to 6 months, and the endochondral ossification was obvious within the drilled bony holes.However, some joints in group B underwent moderate degeneration 12 months after operation. Group C joints in the first month had moderate degeneration, which progressed to severe egeneration 3 months after operation. Conclusion Transplantation of frozen canine phalangeal joint allografts perforated and incorporated with autogenic bone marrow can effectively delay the degeneration of transplanted osteoarticular allografts at the early and middle stage.
Objective To explore an effective method to culture and purify canine bladder transitional epithelial cells.Methods Bladder tissue was obtained from healthy puppy under sterile conditions. Bladder mucosa was removed from the remaining tissue with fine scissor and minced into small pieces, and then were dissociated into single cell suspensions with 0.125% trypsin. The bladder epithelial cells were cultured in defined keratinocyte serum free medium. The cells were passaged and purified by 0.05% trypsin and 0.02% EDTA. Morphological characterization were studied under inverted phase contrast microscope and transmission electron microscope. Expression of cell specific marker protein was assessed by immunohistochemistry. Results Canine bladder transitional epithelial cells could be efficiently cultivated and expanded in serum-free medium without fibroblast contamination. The cells could be passaged 4-6 times without a distinguished decrease in cell proliferation. The cells were characterized by well-developed micro filament and desmosome junction under transmission electron microscope. Immunohistochemical staining with broadly reacting anticytokeratin antibodies (AE1/AE3) confirmed the epithelial phenotype of the cells.Different generations of cells showed diploid cells. Conclusion A large number of bladder transitional epithelial cells can be obtained from small bladder tissue with our digestion method. The cultured bladder epithelial cells can be proliferated to sufficient quantities for further reconstructive purposes.
Objective To observe whether additional penehycl idine hydrochloride (PHC) in mechanical ventilation produces therapeutic effect on oleic acid (OA) induced acute lung injury (ALI) in canine. Methods Seventeen male canines (weighing 12-17 kg) were divided into control group (n=5), OA group (n=6) and PHC group (n=6). ALI model was developed by central venous injection of OA in canines of OA and PHC groups. ALI model was kept steady in air, all groups received mechanical ventilation 90 minutes later. Three groups received normal sal ine 0.25 mg/kg without injection of OA(control group), normal sal ine 0.25 mg/kg after injection of OA (OA group) and PHC 0.25 mg/kg after injection of OA (PHCgroup) respectively at 0 h (90 minutes after onset time of ALI/ARDS). The heart rate (HR), mean arteial pressure (MAP), mean pulmonary arterial pressure (MPAP), central venous pressure (CVP), pulmonary artery wedge pressure (PAWP), artery blood gas analysis, cardiac output (CO), extravascular lung water index (EVLWI), FiO2 and VT were observed respectively at basel ine, onset time of ALI/ARDS and 0 h, then again at 1 hour intervals for 6 hours. Besides the above, airway peak pressure (Ppeak), airway plat pressure (Pplat), mean airway pressure (Pmean) and positve end-expriatory pressure (Peep) were also observed each hour during 1-6 hours. Oxygenation index (OI), pulmonary vascular resistance (PVR), systemic vascular resistance (SVR), alveolar-arterial differences for O2 (AaDO2) and dynamic lung compl iance (DLC) were calculated and pulmonary tissue was collected for histopathologic investigation and dry wet weight ratio (WDR) test. Results The functional parameters of PHC group were improved when compared those of OA group, but there was no siginficant difference; WDR of independent region of three groups were 80.42% ± 3.48%, 82.67% ± 4.01% and 82.26% ± 1.43% respectively; WDR of dependent region of three groups were 80.51% ± 3.60%, 83.71% ± 1.98% and 82.57% ± 1.08% respectively. WDR of PHC group were obviously improved when compared with those of OA group, but there was no significant difference. Independent and dependent regions of PHC group were significantly improved when compared those of OA group in histopathologic scores, alveolar edema, inflammatory infiltration and over-distension (P lt; 0.01). Conclusion Additional PHC in mechanical ventilation produces obvious therapeutic effect on OA induced acute lung injury in canine.
Objective To provide an ideal seed cell for tissue engineered urinary bladder and urethra by serially culturing canine smooth muscle cells from urinary bladder in vitro and compare biological characteristics of different passagesof cells. Methods Bladder smooth muscle cells of 12-month-old male dogs weighing 10-12 kg were isolated from adult dogs’ urinary bladders by collagenase and trypsin digestion and serially cultured in DMEM medium supplemented with 10% serum of newborn bovines. Morphology and prol iferation of the cells were observed and the serially-cultured cells were identified with the transmission electron microscope and immunohistochemistry. Results The cells appeared spindle in parallel rows when they grew to the degree of subconfluence, and showed the “peak-valley” structure under the inverted phase contrast microscope. The cells could be prol iferated serially to the 12th passage in vitro. The growth curve showed the cells before the 7th passage had the similar prol iferation characteristics and the growth cycle was about 40 hours. The TEM showed myofilament and the dense body in cytoplasm of smooth muscle cells. Smooth muscle actin was positive by immunohistochemical staining. After the 7th passage, the cells’ growth became slow, and myofilament and the dense body in cytoplasm vanished. Conclusion The canine smooth muscle cells from urinary bladder can be serially cultured in vitro and highly purified and largely prol iferated by the appropriate method. The cells before the 7th passage can be used as optimal seed cells for tissue engineered urinary bladder and urethra.
目的:分析犬伤暴露后到我院犬伤门诊就诊病例的流行病学相关因素,为制定狂犬病的防治策略提供科学依据。方法:采用描述性统计方法根据犬伤门诊记录,对就诊患者的年龄、性别、发病时间、暴露级别、伤口处理、疫苗接种和犬伤Ⅲ度暴露者狂犬免疫球蛋白的接种及伤人犬只情况等相关因素进行统计。结果:2008年我院犬伤门诊共接诊2 589例,咬伤人犬只的接种率为46.26 %。就诊病例的年龄有两个高峰,分别是0～10岁组和21～30岁组。就诊时间除1～2月和11～12月病例较少外,全年各月份均在198～342人之间。犬伤暴露后,只有2.16 %病例及时、正确地处理了伤口。100%的犬伤暴露者进行了狂犬疫苗的注射。犬伤Ⅲ度暴露中,狂犬免疫球蛋白的使用仅为28.4%。结论:加强犬类动物的管理,对家犬实行免疫,加强对低年龄组儿童的保护。做好狂犬病的防治和知识的宣传工作,合理设置犬伤医学门诊。加大政府投入力度提供全程、免费接种服务。
Objective To evaluate the internal fixation effect, degradation, and biocompatibility of polylactic-co-glycolic acid/hydroxyapatite (PLGA/HA) absorbable cannulated screws in treatment of lateral femoral condyle fracture of canine so as to provide the theory basis for their further improvement and clinical application. Methods Sixteen adult male Beagles (weighing, 9-12 kg) were selected to prepare the models of bilateral lateral femoral condyle fracture; left fracture was fixed with PLGA/HA absorbable cannulated screws as experimental group and right fracture with metal screws as control group. At 2, 4, 8, and 12 weeks after operation, general observation was done and X-ray films were taken for observing fracture healing; bone mineral density was measured; the histological examination was performed; and the degradation property of absorbable cannulated screws was detected. Results All animals survived to the end of the experiment. General observations showed that no fracture displacement occurred and fracture healed at 12 weeks in 2 groups; no breakage, displacement, or loosening of screws was observed in experimental group. X-ray films results showed that the absorbable cannulated screws could not be found out by X-ray in experimental group, but metal screws could be found out in control group; fracture healed with time in 2 groups. The bone mineral density reached the peak at 8 weeks in 2 groups, and no significant difference was found between 2 groups and among different time points in the same group (P gt; 0.05). Histological examination showed that 2 groups had similar fracture healing process at different time points; no obvious inflammatory reaction was found around absorbable cannulated screws in experimental group. The degradation results of absorbable cannulated screws showed that the intrinsic viscosity and molecular weight distribution obviously decreased at 2 weeks; the number average molecular weight and the weight average molecular weight markedly decreased at 4 weeks; and the maximum shear force did not decrease obviously at 8 weeks, and then decreased significantly. Significant differences were found in all indexes among different time points in the same group (P lt; 0.05). Conclusion PLGA/HA absorbable cannulated screws and metal screws show similar fracture healing process for fixing lateral femoral condyle fracture of canine, and the absorbable canulated screws have good biocompatibility. The maximum shear force of PLGA/HA absorbable cannulated screw has no obvious decrease during 8 weeks after operation, so it can ensure full healing of fracture.
Objective Surface modification of nitinol (NiTi) shape memory alloy is an available method to prevent nickel ion release and coating with titanium-niobium (TiNb) alloy will not affect the superelasticity and shape memory of NiTi. To evaluate the bone histocompatibil ity of NiTi shape memory alloy implants coated by TiNb in vivo. Methods NiTi memory alloy columns which were 4 mm in diameter and 12 mm in length were coated with Ti (Ti-coating group) and TiNb alloy (TiNb-coating group) respectively by magnetron sputtering technique. And NiTi group were not coated on the surface. Fifteen mongrel dogs were divided into 3 groups randomly with 5 dogs in each group. NiTi, Ti-coating and TiNb-coating columns were implanted into the lateral femoral cortex of each group, respectively. There were 10 columns embedded in eachdog’s femur whose distance was 1.0 cm to 1.5 cm from each other. The materials were obtained 12 months after operation. After X-ray photography, only those columns which were perpendicular to the cortex of the femur shaft were selected for subsequent analysis. Push-out tests were performed to attain the maximum shear strength (the number of specimens of TiNi group, Ticoating group, and TiNb-coating group were 12, 10, and 14, respectively). Undecalcified sections were used for histological observation and the calculation of osseointegration rate (the number of specimens of TiNi group, Ti-coating group, and TiNb-coating group were 8, 5, and 10, respectively). Results The maximum shear strength of Ti-coating group (95.10 ± 10.03) MPa, and TiNb-coating group (91.20 ± 15.42) MPa were significantly higher than that of NiTi group (71.60 ± 14.24) MPa (P lt; 0.01). Gimesa staining showed that no obvious macrophage and inflammation cell was observed in 3 groups. The osseointegration rates of NiTi group, Ti-coating group, and TiNb-coating group were (21.30% ± 0.23%), (32.50% ± 0.31%), and (38.60% ± 0.58%), respectively; there were significant differences among 3 groups (P lt; 0.01). Conclusion The implants of 3 groups all have good bone histocompatabil ity. But the osseointegration rate and the shear strength in the Ti-coating group and the TiNb-coating group were better than those in the NiTi group, the TiNb-coating group is the best among them.
To evaluate the effect of technique combination of implant-retented titanium lattice with decalcified dental matrix (DDM) implanting. Methods Six healthy male dogs (weighing of 10-20 kg) were randomly divided into 3 groups. All the premolars were extracted on both sides of the jaw in dogs. After 2 weeks, titanium lattice and implant were implanted in the maxillary premolar region with DDM on one side (experimental group), but without on the other side (control group) of each dog. After 4, 9 and 14 weeks, respectively, 2 animals were individually killed each time, and the samples wereevaluated by general observation, X-ray examination, histological observation and histomorphometric analyses. Results General observation: Among the 6 dogs, there was no postoperative infection or death. The X-ray examination showed that the bone density of the experimental group was greater than the control group at 4 and 9 weeks, and had no significant difference as to the vicinity bone at 14 weeks. On the other hand, the density of the control group was very low under the titanium lattice and around the implant. The experimental group revealed a ridge augment of (1.93 ± 0.24) mm, and control group (-1.02 ± 1.20) mm (P lt; 0.05). Developed bone sponge could be found after 14 weeks. Histological observation showed that in the experimental group, the DDM surface was nearly absorbed at 4 weeks. A few new bones were formed at 9 weeks. The whole DDM was absorbed; the trabecular bone was thick and arranged regularly; and the intergradations of implant were observed at 14 weeks. In the control group, there were some inflammatory fibers around the neck of implant at 4 weeks. The inflammatory condition extended to the root of implant and the titanium lattice at 9 weeks. There was no newly-formed bone under the titanium lattice at 14 weeks. Histomorphometric analyses showed that the implant contact bone ratio approached 1 ∶ 1, and showed no significant difference between the new bone fragment and former bone fragment in the experimental group. Conclusion This augmentation of alveolar ridge evaluated by the study is appl icable, but further study is necessary.