Objective To investigate the effect s of T lymphoma invasion and metastasis inducing factor 1 ( Tiam 1) antisense oligonucleotides (ASODN) on morphological remodeling of gast ric cancer cells. Methods The high-invasive and metastastic subgroup (MH ) was separated f rom human gast ric cancer cell line MKN245 (M0 ) by laminin adhesion method in vi t ro. And they were divided into four group s according to different further t reatment s : no t ransfection group (cont rol group ) , liposome t ransfection group , sense oligonucleotides2liposome t ransfection group ( SODN t ransfection with liposome group ) and antisense oligonucleotides2liposome t ransfection group (ASODN t ransfection with liposome group) . Then the expressions of Tiam 1 mRNA and protein were detected by RT-PCR and flowcytomet ry , respectively. The morphology changes between Tima 1 ASODN t ransfected MH cells and no t ransfected cells were observed by using HE stain , cytoskeletal protein stain and scanning elect ronic microscope (SEM) . Results Compared with the other group s , the expressions of Tiam 1 mRNA and protein in MH cells were significantly decreased af ter the cells were t ransfected with 0. 43 μmol/ L Tiam 1 ASODN ( P lt; 0. 01) . Additionally , it was observed that the t ransfected MH cells had less membrane surface projections , fewer or shortener pseudopodia , less irregular cytoskeletal network and less spotted-like actin bodys than no t ransfected MH cells did. Conclusion ASODN t ransfection could effectively suppress the expression of Tiam 1 and the remodeling in gast ric cancer cells , which may play an important role in the invasion and metastasis of gast ric cancer cells.
ObjectiveTo explore the suitable method for isolation and maintenance of primary cultures of human gallbladder epithelial cells (GECs) for establishing the basis of research works in physiological function of gallbladder and its related diseases.MethodsGECs were isolated with collagenase type Ⅳ and blunt separation.The dishes were coated with fibronectin, laminin and polyDlysine respectively.Additional 10 ng/ml epidermal growth factor was added to DMEM medium containing 20% fetal calf serum.The cells were studied under light and electron microscope to determine their shape and distribution.ResultsEach gallbladder yielded approximately (1-5)×107columnar epithelial cells,greater than 95% of which were viable by trypan blue exclusion.The cells grew vigorously within one week which was flat and multangular in shape. CK19 expressed positive.Electron microscope showed typical gallbladder epithelia with microvilli,tight junctions and mucus droplets.ConclusionCombination of collagenase type Ⅳ，mechanical blunt separation and twostep attachment is of great benefit for separating and harvesting GEC.Fibronectin coated culture dish and DMEM medium containing 20% calf serum and 10 ng/ml hEGF is of great benefit for culturing gallbladder epithelial cells.
By using noninvasive venous plethysmography, venography and skin morphology, 44 patients (57 limbs) with chronic venous insufficiency (CVI) in lower extremity were studied , and compared with 12 normal subjects (24 limbs). The results showed that dermal nutrient disturbance caused by deep venous insufficiency accounted for 68%, and followed by perforating venous insufficiency was 44%. Furthermore compared venous refill time (VRT), segmented venous capacitancy (SVC) and maximum venous outflow (MVO) of dermal nutrient disturbance with those of exterior normal skin and normal subjects; and compared VRT, SVC, MVO of deep vein 3-4 stage reflux with those of 1-2 stage reflux and normal subjects,the differences were very significant (Ｐ＜0.05). Compared the VRT of perforating incompetence with that of competence (Ｐ＜0.01). Dermal pathology and ultramicrostructure showed that leucocytes trapping in capillary was a cause of microangiopathy. These results suggest that deep vein 3-4 stage reflux followed by calf perforating insufficiency was a main cause for dermal nutrient disturbance; lower extremity VRT reduced obviously and SVC increased significantly were hemodynamic character, leucocytes trapping in capillary was pathology basis of skin damage.
Objective To investigate the effect of repeated freezing and thawing combining nuclease treatment on the decellularization of bovine tendons, and the morphology, structure, biochemical compositions, and mechanical properties of the decellularized tendons. Methods A total of 48 fresh 1-day-old bovine Achilles tendons were randomly divided into 3 groups (n=16): fresh normal tendons (group A), repeated freezing and thawing for 5 times (liquid nitrogen refrigeration/37℃ thawing, group B), and repeated freezing and thawing combining nuclease processing for 24 hours (group C). In each group, 2 tendons were used for scanning electron microscope (SEM), 3 tendons for histological and immunohistochemical observations, 3 tendons for DNA content detection, and 8 tendons for biomechanical testing. Results SEM observation indicated the intact, aligned, and densely packed collagen fibers with no disruption in groups A and B, and the slightly loose collagen fibers with little disruption in group C. The alcian blue staining, sirius red staining, and immunohistochemical staining showed that the most of glycosaminoglycan, collagen type I, collagen type III, and fibronectin in group C were retained after decellularization treatment. HE and DAPI staining showed that the cell nuclei between the collagen fibers were clearly visible in groups A and B; however, the cell nuclei between collagen fibers almost were invisible with a few residual nuclei on the endotendineum in group C. DNA quantitative detection confirmed that DNA content in group C [(0.05 ± 0.02) μg/mg] was significantly lower than those in group A [(0.24 ± 0.12) μg/mg] and group B [(0.16 ± 0.07) μg/mg] (P lt; 0.05). Biomechanical testing showed that the values of tensile strength, failure strain, stiffness, and elastic modulus were different among 3 groups, but no significant difference was found (P gt; 0.05). Conclusion Repeated freezing and thawing combining nuclease processing can effectively remove the component of cells, and simultaneously retain the original collagen fibrous structure, morphology, most of the extracellular matrix compositions, and mechanical properties of the bovine tendons.
Objective To investigate the morphological anatomical abnormal ities of high congenital dislocation of hip in adults and provide anatomical basis for the total hip arthroplasty (THA). Methods From May 1997 to July 2008, 49 patients (57 hi ps) with high congenital dislocation of hip (Hartofilakidis type III) were treated. There were 6 males and 43 females with an average age of 29.4 years old (18-56 years old). The locations were left in 24 hi ps and right in 33 hi ps. The morphological parameters (including femoral length, isthmus, height of femoral head center, neck-shaft angle, medialhead offset, anteversion angle, canal flare index, anteroposterior diameter of the true acetabulum, posterior thickness of the true acetabulum, depth of the true acetabulum) of suffering hips (dislocation group, n=57) were measured by preoperative X-ray, CT and intraoperative cl inical observation and were compared with those of contralateral hips (control group, n=41). The intraoperative situations of hip were observed. Results The height of dislocation was (45.41 ± 2.15) mm. The length difference of both lower extremities was (40.41 ± 2.02) mm. In dislocation group, isthmus was shortened; height of femoral head center, neck-shaft angle and medial head offset were decreased; and anteversion angle was increased. CT showed that the canal flare index was larger than 4.7, femoral shape was funnel-shaped according to Noble classification. Anteroposterior diameter of the true acetabulum became smaller, posterior thickness of the true acetabulum became thicker, and depth of the true acetabulum was shallower. There were statistically significant differences in the morphological parameters of femur and acetabulum between two groups (P lt; 0.05). The intraoperative measurements showed that the anteroposterior diameter of acetabulum was (32.98 ± 1.02) mm and the depth of acetabulum was (14.21 ± 0.56) mm. There was no statistically significant difference between intraoperative measurements and preoperative measurements (P gt; 0.05). The acetabulum was full of fat and fibrous tissues. Running of the sciatic nerve in 40 cases were changed and it ran upward and laterally. Conclusion When high congenital dislocation of the hip in adults is treated with THA, anatomical variation must be fully taken into account. The acetabulum is expanded toward posterosuperior, excessive reamed should be avoided to prevent femoral fractures, and appropriate or tailor-made prosthesis was selected.
【Abstract】 Objective To explore the methods and appl ication value of surface shaded display (SSD) and multiplanarreconstruction (MPR) in the evaluation of acetabular morphology in patients with developmental dysplasia of the hip (DDH) before total hip arthroplasty (THA). Methods From October 2003 to November 2006, 17 patients (3 males and 14 females, aging from 35 years to 61 years) with osteoarthritis secondary to DDH were scanned with spiral CT preoperatively. According to the Crowe standard, 19 dysplasia hips were classified as type I in 4 hips, type II in 9 hips, type III in 6 hips. The obtained hip CT data were developed with SSD and MPR to observe spational position and bone stock of the acetabula. Results The dislocated extent was 25%-89% in these dysplasia hips according to the Crowe method and their sharp angles all exceeded 45°. Bone defect occurred to each of the acetabula, among which it was located in anterosuperior acetabulum in 5 hips, in superolateral acetabulum in 11 hips and in posterosuperior acetabulum in 3 hips. The hip images made with MPR showed that the minimum thickness of the medial wall of acetabula ranged from 2.0 mm to 10.9 mm. Among 15 unilateral dysplasia patients, the opening difference anddepth difference between the dysplasia acetabulum and the contralateral one ranged from 2.7 mm to 19.1 mm and from 2.3 mm to 13.1 mm, respectively. Conclusion SSD and MPR of spiral CT are effective methods in evaluating acetabular morphology preoperation and contribute to intraoperative acetabular reconstruction in patients with DDH performed THA.
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 investigate the morphological changes of the proximalfemur and their implication to the total hip arthroplasty in patients with Crowe Ⅱ/Ⅲ developmental dysplasia of the hip (DDH). Methods The experimental gr oup was composed of 15 hips in 14 patients (Crowe Ⅱ, 9 hips; Crowe Ⅲ, 6 hips ) with osteoarthritis secondary to Crowe Ⅱ/Ⅲ DDH (2 males, 12 females; age, 35-61 years). None of the patients had accepted any osteotomy treatment. The control group was composed of 15 normal hips in 15 patients with unilateral DDH (3 males, 12 females; age, 35-57 years). Twelve hips came from the experimental group and the other 3 came from the patients with unilateral Crowe Ⅰ DDH. The femurswere examined with the CT scanning. The following parameters were measured: theheight of the center of the femoral head (HCFH), the isthmus position (IP), theneckshaft angle(NS), the anteversion angle, the canal flare index, and the canal width. Then, the analysis of the data was conducted. Results HCFH and IP in theexperimental group and the control group were 50.1±6.7 mm, 50.1±7.4 mm, and 107.4±21.5 mm, 108.7±18.1 mm，respectively, which had no significant differencebetween the two groups（Pgt;0.05）. In the experimental group and the control group, the NS were 138.3±10.0° and 126.7±5.7°，the anteversion angles were 36.5±15.9° and 18.8±5.4°, and the canal flare indexes were 4.47±0.40and 5.01±0.43. There was a significant difference between the two groups in the above 3 parameters （Plt;0.05）. As for the canal width of the femur, therewasa significant difference in the interior/exterior widths and the anterior/posterior widths at the level of 2 cm above the lesser trochanter and 4 cm belowthe lesser trochanter between the two groups （Plt;0.05）; however, there was nosignificant difference in the canal width of the femur at the isthmus between the two groups（P＞0.05）. Conclusion It is necessary to evaluate the morphology of the proximal femur before the total hip arthroplasty performed in patients with Crowe Ⅱ/Ⅲ DDH. The straight and smaller femoral prosthesis should be chosen and implanted in the proper anteversion position duringoperation.
Objective To investigate the effect of the morphological changes in the proximal femur on the prothesis selection in the total hip arthroplasty in the patients with ankylosing spondylitis. Methods The experimental group was composed of 13 patients (16 hips) with ankylosing spondylitis, which was treated with the total hip arthroplasty, and the control group was composed of 16 patients（19 hips）with non-ankylosing spondylitis，which was also treated with the total hip arthroplasty. In the two groups, the measurements of Singh index，canal flare index，morphological index of the cortex and cortical index were performed in the two groups. Results The results of the statistical analysis on Singh index，canal flare index, morphological index of the cortex and cortical index in the experimental group were 3.81±0.54, 2.63±0.41, 2.02±0.38 and 1.69±0.69, respectively, but 4.63±0.62, 3.03±0.27, 2.76±0.28 and 2.12±0.24, respectively in the control group. Therewas a significant difference in Singh index, canal flare index, and morphological index of the cortex between the two groups (Plt;0.05)，while there was no statistical difference in cortical index between the two groups (P＞0.05). The patientswith ankylosing spondylitis had more serious osteoporosis in their proximal femur. Conclusion Cemented femoral prosthesis should be used in the total hip replacement in patients with ankylosing spondylitis, and the revision total hip arthroplasty should be performed on patients with more serious osteoporosis.
Objective To study the healing ability of the central area tissue of flexor tendons after injury. Methods Tendons of flexor digitorum profundus of the long toes from 8 white Leghorn hens were harvested in zone II. Tissues were cut in 4 mm segments and divided into the experimental group(the central area tissue of flexor tendons) and the control group(the tendon segments without epitenon). There were 12 tendon segments cultured in each group. Specimens were obtained and examined under light microscope on the 9th, 18th and 27th days after culture, respectively. Another 4 tendons were used as normal control, and they were directly examined under light microscope. Results The number of tenocytes was significantly less in the control group than in the experimental group and the uncultured state (P＜0.01); the number of tenocytes was significantly higher in the experimental group than in the uncultured state (P＜0.01). The number of tenocytes of the experimental group were higher on the 9th day than on the 18th and 27th days after culture(P＜0.01). Conclusion The central area tissue of flexor tendons has favorable healing ability after injury.