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 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 prepare chitosan-gelatin/hydroxyapatite (CS-Gel/HA) composite scaffolds, and to investigate the influence of components and preparing conditions to their micromorphology. METHODS: The CS-Gel/HA composite scaffolds were prepared by phase-separation method. Micromorphology and porosity were detected by using scanning electron microscope and liquid displacement method respectively. RESULTS: Porous CS-Gel/HA composite scaffolds could be prepared by phase-separation method, and their density and porosity could be controlled by adjusting components and quenching temperature. CONCLUSION: The study suggests the feasibility of using CS-Gel/HA composite scaffolds for the transplantation of autogenous osteoblasts to regenerate bone tissue.
ObjectiveTo investigate the cytotoxicity of triamcinolone acetonide (TA) on cultured human retinal pigment epithelial (RPE) cells.MethodsEffect of TA with different concentraion (0.4, 02, 01, 0.05, and 0.025 mg/ml) on the proliferation of RPE cells was detected by methyl thiazolyl tetrazolial (MTT) assay; The changes of cellular cycle treated by TA with the drug concentration of IC50 for 72 hours were measured by flow cytometry (FCM) ananlysis, and the morphological and ultrastructural changes of the cells were observed by phasecontrast microscopy and transmission electron microscopy.ResultsWith the concentration of 0.4-0.025 mg/ml, TA inhibited the growth of RPE cells obviously in a dose-dependent manner compared with the control (P＜0.05), and the cells in S phase treated with TA decreased 10.6% compared with the control (P＜0.05). Under the light microscope, sparse cells with irregular configuration and many prominences and cellular vacuoles in TA group can be seen. The results of transmission electron microscopy showed asymmetrically distributed cellular chromatin was, cavitated cytoplasm, and even some necrotic cells.ConclusionTA can inhibite the S-phase karyomitosis of RPE and inhibite the cellular proliferation; and destroye the cellular structure and lead to the necrosis, which indicates the cytotoxicity of TA on RPE.(Chin J Ocul Fundus Dis, 2005,21:233-236)
ObjectiveTo investigate the anatomical morphology of the proximal femur in adult patients with Crowe type IV developmental dysplasia of the hip (DDH) so as to provided a reference for the selection of femoral components or personalized artificial prosthesis for DDH. MethodsBetween June 2004 and December 2014, 49 patients (58 hips) with Crowe type IV DDH were included in this study. There were 7 males (8 hips) with an average age of 47.3 years and 42 females (50 hips) with an average age of 30.0 years. X-ray films were taken in all cases and CT scanning in 17 cases (17 hips). The anatomical parameters were measured through Osirix V 5.8.5 software. ResultsThe distance of femoral head dislocation was (6.09±1.04) cm;the femoral head and neck disappeared completely in 8 hips (13.8%);femoral head atrophy and deformation and femoral neck disappearance were observed in 11 hips (19.0%). The midpoint width of the small trochanteric canal was (1.53±0.29) cm;the isthmus diameter was (0.69±0.09) cm;the canal flare index (CFI) value was (3.50±0.42) cm;the femoral neck anteversion angle was (37.0±18.2)°;and all indexes showed significant difference when compared with ones of normal population and Crowe type I, II, and III patients. But there was no significant difference in femoral neck-shaft angle (132.3±9.1°), isthmus height (11.66±2.67) cm (P<0.05). The chimney type cavity (CFI<3) accounted for 51.7%, which was significantly higher than the value of normal population. There was a positive correlation between metaphyseal CFI and CFI (r=0.63, P=0.00), but there was no correlation between metaphyseal CFI and distal CFI (r=-0.17, P=0.21). ConclusionThe proximal femur has the morphological characteristics of narrow medullary cavity, increased anteversion angle, and high ratio of chimney type cavity, so it is necessary to design specialized femoral components or personalized artificial prosthesis for the DDH.