OBJECTIVE: To study the characteristics of, morphology histology and ultrastructure of anterior cruciate ligament(ACL) autograft and two-step cryopreserved ACL allograft after transplantation. METHODS: Sixty New Zealand rabbits and sixty Japanese rabbits were randomly divided into two groups: ACL autograft group and two-step cryopreserved ACL allograft group. Immunosuppressant were not used after transplantation. The histology and ultrastructure of the ACL of transplantation and normal knee were observed after 4 weeks and 12 weeks, respectively. RESULTS: The rate of remodeling process was faster in ACL autograft than in two-step cryopreserved ACL allograft, but there was similar remodeling process between two groups 12 weeks after transplantation. The proportions of large-diameter fibers(gt; or = 80 nm) of ACL autograft and cryopreserved ACL allograft were 6% and 24% in the 4th week, and were 0 and 2% in the 12th week, respectively. The proportions of small-diameter of fibers(lt; 80 nm) of ACL autogrft and cryopreserved ACL allograft were 94% and 76% in the 4th week, and 100% and 98% in the 12th week, respectively. Histologic incorporation in ACL autograft was similar to that in cryopreserved ACL allograft. CONCLUSION: Two-step cryopreserved bone-ACL-bone allograft were similar to bone-ACL-bone autograft cryopreserved in remodeling process and histology. The rate of remodeling process was faster in ACL autograft than in cryopreserved ACL allograft.
Objective To study ultrastructure and clinical significance of gastrin secretory granule in colorectal carcinoma cells. Methods The gastrin expression in colorectal carcinoma tissue and blood of 10 cases was examined by using radioimmunity analysis and immunohistochemistry. The ultrastructure of gastrin secretory granule of 10 cases, the positive of gastrin immunohistochemistry of colorectal carcinoma were examined by using immunoelectron microscopic technique. Results The gastrin concentration of the colorectal cancer group 〔（130.75 ±21.34） pg/ml〕 was significantly higher than that of control group 〔（95.63± 12.26） pg/ml〕，Plt;0.05. In 10 specimens of colorectal cancer, 5 cases were gastrin immunohistochemistry positive (+++), 4 moderate positive (++) and 1 weak positive (+). Cells in colorectal cancer were polyshaped, with unusual nucleoli different in size, concentrating on the edge, the cytoplasm mitochondrion was plentiful with vacuolates, and more secretion granules could be seen, 400-1500 nm in diameter with a clear border of membrane. There were two types of granular appearance: type A was largest in bulk size, low electrodensity was welldistributed, granular core appeared loose; type B was smaller in bulk size, high electrodensity was welldistributed, nucleus was usually compact.protein A gold (pAg) positive granules were located partially in secreting granules. pAg positive granules in highly differentiated cancer were mainly located in secreting granules of type A. pAg positive granules in low differentiated cancer were mainly located in secreting granules of type B. A part of cancer cell membrane, and inside and outside of microvillus membrane, adhering to pAg granules in line could be seen. Conclusion The colorectal carcinoma cells may synthesize and secrete gastrin themselves, which may be the mechanism of high gastrin levels in colorectal cancer. The use of gastrin antagonist and receptor antagonist may treat the patents with colorectal carcinoma.
Objective To observe the relationship of osteoblasts, endothelial cells and ceramic scaffold during reconstruction of rat critical size calvarial defects with tissue engineering technique under transmission electron microscope. Methods Fourteen male adult Sprague Dawley rats were divided randomly into experimental and control groups. Bone marrow was obtained from left femurs and tibias of all rats. In experimental group, respective autogenous osteoblasts derived from bone marrow stromal cells(MSCs) different iated and proliferated in vitro and then were seeded and subcultured on porous calcium phosphate ceramics. The cell-ceramic compounds were used to repair critical-sized (8 mm diameter) calvarial defects in the corresponding rats. In control group, the ceramic without autogenous osteoblosts was used. One rat of each group was sacrificed postoperatively in the 4th, 8th, 12th, 24th, 28th weeks respectively and involved samples were removed to make decalcified ultrath in sections and observed under transmissionelectron microscope. Results Osteoblasts or osteoblast-like cells always located next to sprouting capillaries and the relationship between osteoblasts and endothelial cells was relevant in experimental group. There was a calcium depositzone distributed along the boundary of newly formed bone and the remnants of decalcified ceramic, which meant osseointegration between the ceramic and newly formed bone. The above changes did not appear in control group simultaneously.Conclusion The nanometer scale structure of ceramic scaffold benefits to angiogenesis, osteogenesis and extracellular matrix formation in repair bone defects with tissue engineering technique.
Objective To observe ultrastructural changes of the intervertebraldisk in the corresponding area after internal fixation of spinal column. Methods Twenty-four Japanese big ear rabbits were divided into internal fixation of spinal column group (n=12) and control group (n=12). The internal fixation model was made as follows: The spinous processes and erector spinal muscle were exposed and the T10L3 spinous processes and the relevant two-side articular processes under the periosteumwere isolated. With the help of L-shaped Kirschner wires, the steel wire was threaded through the articular of T11,T12,L1 and L2, and were connected with L-shaped Kirschner wries. After 6 months of operation, the following intervertebral disk tissues were observed with transmission electeon microscope: nucleus pulposus, internal annlus fibrosus and external anulus fibrosus of L1 intervertebraldisk. The T12and L2 intervertebal disk surface structure was observedhorizontally and longitudinally with scanning electron microscope, respectively. Results After internal fixation of spinal column, the structural changes of cells in nucleus pulposus and internal annulus fibrosus occurred earlier than that in the external annulus fibrosus. Proteoglycan and special structure were found in nucleus pulposus and matix of annulus fibrosus. However, the forms of special structure in nucleus pulposus and internal layer of annulus fibrosus were different. In the degeneration matrix of intervertebral disc, the proteoglycan particles and special structure were obviously decreased. Conclusion Abnormal stress environment can result in the degeneration of intervertebral disk. There is a regular distribution of the special structure in nucleus pulposus and matrix of annulus fibrosus, which is related to biology behaviour of proteoglycan particles in the degeneration of intervertebral disk.
ObjectiveTo observe the ultrastructural changes of vasa vasorum endothelial cells in the walls of the great saphenous vein and splenic vein, and to evaluate the effect of high hydrostatic pressure and hypoxia upon vasa vasorum endothelial cells.
MethodsThirty-four varicose great saphenous vein samples and splenic vein samples with portal hypertension were obtained, and the same number of normal great saphenous vein and splenic vein were used as the control groups. Semi-thin sections stained with HE staining vasa vasorum of the adventitia in great saphenous vein and splenic vein were observed for light microscopy. Samples were made into ultrathin-slices again. The ultrastructural changes of endothelial cells were observed under transmission electron microscopy.
ResultsIn varicose great saphenous veins and diseased splenic veins, the nuclear architecture of endothelial cells in vasa vasorum were integrity and the distribution of chromatin were normal. In some mitochondria, the trachychromatic groundplasm, undefined and ruptured cristae were found.
ConclusionUnder high hydrostatic pressure and hypoxia conditions, the ultrastructure of vasa vasorum endothelial cells between the great saphenous vein and the splenic vein may appear remodeling phenomenon, and both changes are similar.
In order to study the influence of biliary tract obstruction on enteromicroflora,we ligate the canine biliary tract to observe the acrobic and anerobic bacteria in the duodenum and ileum at intervals of post-ligation(the 10th,20th,30th days),and to study the pathogenesis and ultramicroscopic of the ileal mucosa at the same intervals.The results showed that:the population and species of enteroflora in small intestine gradually increased after biliary obstruction.Bacteria(especialy E.coli) ascended to the upper part of small intestine,from their normal habitant of lower part of small intestine.Therefore the radio of general aerobia and E.coli risen obviously in duodenum.The longer the obstruction,the more pathologic changes were observed in ileal mucosa.such as edema,leukocytes infiltration and destruction of epithelial villi.All of those changed may be the causative factor of biliary tract infection.So that,in the programs of preventing enterogenic infection at the state of biliary tract obstruction,the protection and adjusting of normal enteroflora should be adventently considered.
Objective To investigate whether the agonist of delta opoid receptor D-Ala(2),D-Leu(5) enkephalin (DADLE) has the effect of decreasing myocardial injury during ischemia-reperfusion of adult rabbits’ myocardium，so that a new mehanism and way to myocardial protection could be found. Methods Langendorff model was used during the experiment. Thirty rabbits were divided into three groups randomly (each group 10 rabbits). Control group: St.Thomas Ⅱ cardioplegic solution was used; group 1: St.Thomas Ⅱ cardioplegic solution and DADLE (1mg/kg) were used; group 2: St.Thomas Ⅱ cardioplegic solution and naloxone(3mg/kg) were used to induce the hearts to arrest respectively. After arrest the hearts were reperfused respectively. Data of left ventricle development pressure(LVDP) was recorded before and after ischemia. Biochemical indicators of myocardium, lactate dehydrogenase(LDH) were detected before and after ischemia. Some myocardial tissues were used to explore the changes of the tissue of ultrastructure with electron microscope，when the experiment was over. Still some myocardial tissues were to be detected by flow cytometer to evaluate the apoptosis of the myocardium. Results The LDH and LVDP showed significant difference among three groups after ischemia(Plt;0.05); LVDP in group 1 was higher than those in group 2 and control group(69.8±5.8 mmHg vs. 23.4±3.9 mmHg; 69.8±5.8 mmHg vs. 37.9±4.7 mmHg; Plt;0.05), the LDH in group 1 was lower than those in group 2 and control group(1 272.6±59.1 U/L vs. 2 764.4±27.7 U/L, 1 272.6±59.1 U/L vs. 1 884.4±37.5 U/L; Plt;0.05). The apoptosis rate in group 1 was lower than those in group 2 and control group. As could be shown from the ultrastructure: mitochondria structure was nearly normal in group 1; mitochondria structure was injuried severely in group 2; there was a minor injury in control group. Conclusion Agonist of δ opoid receptor DADLE in cardioplegic solution could induce hibernation, which has myocardial protection effect during ischemia-reperfusion injury.
Polypoidal choroidal vasculopathy (PCV) is originally defined as a separate disease, but with the development of imaging techniques, it has now been included in the spectrum of neovascularization. In the Asian population, the prevalence of PCV is high, and with the deepening of clinical studies, the pathological characteristics, pathogenesis and clinical manifestations of PCV have been more deeply understood. Through dynamic observation and histopathological study of PCV lesions during operation, it can be confirmed that the lesions are mainly located between the retinal pigment epithelium and the Bruch membrane, rather than originating from the choroidal circulation, which is of great significance for understanding the origin and natural course of PCV. It is worth noting that although a theoretical bridge has been established between age-related macular degeneration (AMD)/PCV, there is a lack of intuitive clinical data on the ultrastructural and molecular manifestations of the cells/stroma in the local lesions of the eye, especially the progression of AMD/PCV from early/middle stage to exudative stage. It is precisely because of this that highly attractive research topics and exploration space are proposed for the future.
Objective To explore the ultrastructure characteristics of pulmonary arteries in smokers with normal lung function and with chronic obstructive pulmonary disease ( COPD) . Methods 33 patients who undertook surgery for peripheral lung cancer were collected. According to smoking history and pulmonary function, the patients were divided into three groups, ie. non-smokers with normal pulmonary function ( group A, n = 10) , smokers with normal pulmonary function ( group B, n = 13) , and smokers in stable phase of COPD ( group C, n = 10) . Normal lung tissues without cancer were sampled and observed under light and electric microscope. Results ①Compared with group A, the thickness of intimal layer of intra-acinar pulmonary muscular arteries of group B and C were significantly higher, the area of their lumenwas lower, and the proportion of their muscular arteries was higher( P lt; 0. 01) . ②Ultrastructure of small pulmonary arteries of group A showed that intimal layer was normal, so as to endothelial cells and smooth muscle cells. Collagen fiber was not increased. Ultrastructure observation of group B showed that endothelialcells were distorted, basal membrane was thick, and collagen fiber increased in vessels. Ultrastructure observation of group C showed that endothelial cells degenerated, vascular intima thickness increased, andsynthetic phenotype smooth muscle cells increased. ③ Smoking index was positively correlated with the proportion of muscular arteries and the proportion of intimal area( r =0. 464,0. 635, P lt;0. 05, respectively) ,and negatively correlated with the proportion of lumen area( r= - 0. 603, P lt;0. 05) . Conclusions Smokers with normal lung fuction and with COPD show the similar ultrastructural characterizations in endothelial cells, smooth muscle cells, and pulmonary arterial remodeling, which related closely to smoking.
In order to investigate the effect of nerve compression on neurons, the commonly used model of chronic nerve compression was produced in 48 SD rats. The rats were sacrificed in 1, 2, 3, 4, 5 and 6 months after compression, respectively. The number of neuron and ultrashruchure of alpha-motor neurons and ganglion cells of the corresponding spinal segment were examined. The results showed as following: After the sciatic nerve were crushed, the number of neuron and ultrastructure of alpha-motor neurons and ganglion cells might undergo ultrastructural changes, and even the death might occur. These changes might be aggravated as the time of crushing was prolonged and the compression force was increased. It was concluded that for nerve compression, decompression should be done as early as possible in order to avoid or minimize the ultructural changes of the neuron.
