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.
Abstract： Objective To investigate the early and mid-term outcomes of morphologic tricuspid valve replacement by means of intravalvular implantation in corrected transposition of great arteries（cTGA）. Methods From January 2009 to January 2012，11 patients with cTGA were surgically treated in Fu Wai Hospital. There were 9 male patients and 2 female patients with their mean of age of（37.8±11.7）years and mean body weight of（73.0±11.3）kg. All the patients underwent morphologic tricuspid valve replacement with preservation of the entire valvular and subvalvular apparatus. Simultaneous surgical procedures included repair of ventricular septal defect in 2 patients，repair of atrial septal defect in 4 patients，pulmonary valvuloplasty in 1 patient，reconstruction of functional right ventricular outflow tract in 4 patients and repair of coronary-pulmonary artery fistula in 1 patient. Postoperative New York Heart Association （NYHA） classification, cardiothoracic ratio, morphological right ventricle ejection fraction, end-diastolic dimension of morphological right ventricle and left atrium were evaluated during follow-up. Results All the 11 patients were successfully surgically treated and followed up for an average duration of（13.0±10.6）months. There was no statistical difference between postoperative and preoperative average cardiothoracic ratio （0.54±0.06 vs. 0.57±0.09，t=1.581，P＞0.05），morphologic right ventricle ejection fraction （52.8%±9.0% vs. 54.9%±9.5%, t =0.712，P＞0.05），and end-diastolic dimension of . morphological right ventricle （54.3±7.5 mm vs. 56.9±9.2 mm，t =0.988，P＞0.05）. There was statistical difference between postoperative and preoperative average end-diastolic dimension of left atrium（42.1±8.9 mm vs. 53.4±11.1 mm，t =3.286，P＜0.05）and NYHA classification（Z = -2.640，P＜0.05）. Conclusion Intravalvular implantation of morphologic tricuspid prosthesis can protect the physiological structure of morphologic right ventricular and prevent furtherdamage to its function caused by morphologic tricuspid valve insufficiency. Postoperative dimension of morphologic left atrium and cardiac function are significantly improved. The early and mid-term outcomes are satisfactory．
Objective To investigate the surgical indications and the mid and long term results of morphologic tricuspid valve replacement for corrected transposition of the great arteries（cTGA）. Methods From September 1997 to September 2007, 18 cases with cTGA were treated in Fu Wai Hospital. There were 15 male and 3 female, aged from 16 to 51 years（33.3±12.8 years）, and weighed from 47 to 90 kg(60.9±14.7 kg). There were 10 cases with isolated morphologic tricuspid valve insufficiency, 3 complicated with ventricular septal defect, 2 complicated with ventricular septal defect and pulmonary valve stenosis, 2 with morphologic tricuspid valve insufficiency after septal defect repair, and 1 with mechanical valve dysfunction after morphologic tricuspid valve replacement. The preoperative mean morphologic right ventricle ejection fraction was 562%±11.6%. Of the 18 cases, 12 were in grade Ⅱ and 6 were in grade Ⅲ according to New York Heart classification（NYHA）.All the cases had undergone morphologic tricuspid valve replacement. Postoperative indices such as cardiac function and morphological right ventricle ejection fraction were followed up. Results One patient died of postoperative low cardiac output syndrome. Two had pervavlvular leak, which were cured by pervavlvular leak repair at 7th and 30th day after operation, respectively. Sixteen were followed up with a followup time of 57.0±407 months. There was no statistical significance between preoperative and postoperative mean morphologic right ventricle ejection fraction（52.8%±9.2% vs.56.2%±11.6%; t=2.062, Pgt;0.05）. The followup showed that 12 were in NYHA grade Ⅰ or Ⅱ, and 4 were in NYHA grade Ⅲ. There was no statistical significance between preoperative and postoperative percentage of cases in NYHA grade Ⅲ（χ2=1.532，Pgt;0.05）. Conclusion Morphologic tricuspid valve replacement can prevent the further damage to morphologic right ventricular function caused by morphologic tricuspid valve insufficiency. The mid and long term results were satisfying. During the followup, the morphologic right ventricle can function appropriately.
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 changes of lumbar bone histomorphometry after exposure to low frequency pulsed electromagnetic fields (PEMFs), and to further understand the effect of PEMFs on osteoporosis (OP) in ovariectomizedOP rats. Methods Sixty-six 3-month-old Sprague Dawley rats were randomly divided into 4 groups: group A(n=12), groupB (n=12), group C (n=12), and group D (n=30). In group A, the ovaries were not resected as sham-ovariectomy; in groupsB, C, and D, the ovaries were resected. At 12 weeks after ovariectomy, the rats were exposed to PEMFs at 8 Hz, 3.8 mT, and 40 minutes/ day for 30 days in group B; the rats were administered with premarin [0.065 mg/(kg·d) by gavage for 30 days] in group C; in group D, the rats were housed as ovariectomy control. The hair and activity of rats were observed; the levels of serum estradiol were determined. At 30 days after intervention, all rats were sacrificed to harvest the L4 vertebrae for bone histomorphometry. Results General observation showed hair loss and decreased activity in group D, and no abnormal appearances in groups A, B, and C. The level of serum estradiol in group A was significantly higher than that in group D [(54.93 ± 23.52) pg/mL vs. (31.99 ± 23.45) pg/mL] (t=2.345, P=0.029). Histological observation showed thinness of sclerotin, bigger medullary cavity, and sparse and thinner bone trabecula in group D; uniform bone trabecula with no breakage in groups A, B, and C at 30 days after intervention. The ratio of trabecular bone area in group B was significantly higher than that in group D (P lt; 0.05); it was higher than that in groups A and C, showing no significant difference (P gt; 0.05). The trabecular thickness in group B was significantly higher than that in group D (P lt; 0.05), but it was lower than that in groups A and C, showing no significant difference (P gt; 0.05). The trabecular number in group B was significantly lower than that in group D (P lt; 0.05), but it was higher than that in groups A and C, showing no significant difference (P gt; 0.05). The trabecular separation in group B was higher than that in group D and lower than that in groups A and C, showing no significant difference (P gt; 0.05). Conclusion PEMFs at 8 Hz and 3.8 mT can significantly improve the character of bone microstructure in ovariectomized OP rats, increase the ratio of bone trabecular area and trabecular thickness, and decrease the trabecular number.