Application of Hanging Drop Technique for Kidney Tissue Culture.

BACKGROUND/AIMS: The hanging drop technique is a well-established method used in culture of animal tissues. However, this method has not been used in adult kidney tissue culture yet. This study was to explore the feasibility of using this technique for culturing adult kidney cortex to study the time course of RNA viability in the tubules and vasculature, as well as the tissue structural integrity. METHODS: In each Petri dish with the plate covered with sterile buffer, a section of mouse renal cortex was cultured within a drop of DMEM culture medium on the inner surface of the lip facing downward. The tissue were then harvested at each specific time points for Real-time PCR analysis and histological studies. RESULTS: The results showed that the mRNA level of most Na+ related transporters and cotransporters were stably maintained within 6 hours in culture, and that the mRNA level of most receptors found in the vasculature and glomeruli were stably maintained for up to 9 days in culture. Paraffin sections of the cultured renal cortex indicated that the tubules began to lose tubular integrity after 6 hours, but the glomeruli and vasculatures were still recognizable up to 9 days in culture. CONCLUSIONS: We concluded that adult kidney tissue culture by hanging drop method can be used to study gene expressions in vasculature and glomeruli.

Unicondylar osteoarticular allograft reconstruction of the distal femur in a patient with a traumatic osteoaticular defect of the lateral femoral condyle.

PURPOSE: The objective of this study was to report the clinical results for a patient with a post-traumatic osteoarticular defect of the lateral femoral condyle treated with unicondylar osteoarticular allograft reconstruction. METHOD: We describe the case with a massive osteoarticular loss of the lateral femoral condyle combined with a grade IIIA open distal femoral diaphyesal fracture. The patient underwent unicondylar osteoarticular reconstruction with an allogenic fresh-frozen condyle. RESULTS: There were no complications after the reconstructive procedure. At the five year follow-up, the patient showed a favorable clinical outcome with a full range of motion of the knee joint. Degenerative changes were seen in the graft compartment, but they did not cause serious functional limitations. CONCLUSION: Unicondylar allograft reconstruction appears to be a surgical option for patients who suffer from a post-traumatic distal femoral osteoarticular defect in which only one condyle is absent or severely damaged.

Total hip arthroplasty for treatment of fused hip with 90 degrees flexion deformity.

Conversion of a fused hip to total hip arthroplasty is a challenging procedure. The results depend on the degree of the change that occurs in the muscles and soft tissues at the hip from long immobilization, and considerable postoperative rehabilitation is necessary. Complications such as nerve injury, dislocation, infection and heterotopic ossification are common. Most of the authors recommended trochanteric osteotomy for better exposure. We are reporting a case of conversion total hip arthroplasty performed for a hip fused in 90 degrees of flexion. Surgical technique is described in detail and pertinent literature is reviewed.

Bilateral simultaneous two-incision minimally invasive total hip arthroplasty.

BACKGROUND: This study aimed to assess the feasibility of bilateral simultaneous minimally invasive two-incision total hip arthroplasty in terms of clinical and radiological outcomes, complications, and patient satisfaction. METHODS: A total of 62 patients, ages 24-69 years, were operated on with bilateral simultaneous minimally invasive total hip arthroplasty using a modified two-incision technique, after which they were followed for an average 41 months. The acetabular component was inserted through part of the Watson-Jones approach, and a femoral component was inserted through a space between the gluteus medius and piriformis tendon. RESULTS: The mean duration of surgery was 180.4 min, and the mean perioperative blood loss was 1513.2 ml. There were no intraoperative complications. Postoperative periprosthetic fracture occurred in two patients and delayed infection in one patient. The average Harris Hip Score improved from 41.8 to 95.3. The WOMAC score improved from an average of 66.2 to 5.0. The average lateral opening angle of the acetabulum was 40 degrees , and the angle of anteversion was 12 degrees . All the femoral components were implanted in neutral to 5 degrees valgus position. CONCLUSIONS: Bilateral simultaneous minimally invasive total hip arthroplasty using a modified two-incision technique gave satisfactory clinical, radiological, and functional results. Efforts should be made to prevent perioperative periprosthetic fractures.

Interaction between malignant transformation of human pulmonary epithelial cells and activation of fibroblasts induced by Yunnan tin mine dust / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the interaction between transformation of human pulmonary epithelial cells and activation of fibroblasts induced by Yunnan tin mine dust.</p><p><b>METHODS</b>(1) The immortalized human bronchial epithelial cell line BEAS-2B and human embryo lung fibroblast cell line WI-38 were grown in MEM medium containing 5% and 10% FBS, respectively, at 37 degrees C and 5% CO2 with saturated humidity. The cells were subcultured every 6 days. BEAS-2B cells and WI-38 cells were induced with Yunnan tin mine dust on every other generation at the concentration of 100 microg/ml. From the 11th generation, the cells were co-cultured. Epithelial cell transformation was tested using concanavalin A (ConA) agglutination and anchorage-independent growth assays. The cell cycles were analyzed through flow cytometry. The expressions of alpha-SMA in fibroblasts were determined with immunocytochemistry.</p><p><b>RESULTS</b>(1) Cell morphology of mine dust-exposed epithelial cells began to transform at the 28th generation. Similar transformations were observed with mine dust-induced epithelial cells co-cultured with fibroblasts from the 20th generation and mine dust-induce epithelial cells co-cultured with mine dust-induced fibroblasts from the 16th generation. ConA agglutination assay and anchorage-independent growth assays were negative in normal BEAS-2B cells. At the 26 th generation, the agglutination test result of the mine dust-exposed epithelial cells was positive. Co-cultured with fibroblasts and mine dust-exposed fibroblasts, the agglutination time of the mine dust-exposed epithelial cells became short. Epithelial cell anchorage-independent growth assay was positive for mine dust-exposed epithelial cells co-cultured with fibroblasts at the 36th generations and for mine dust-exposed epithelial cells co-cultured with mine dust-exposed fibroblasts at the 26th generations. The clone formation rate of the 26th generation was 6.00 per thousand +/- 1.00 per thousand and 15.33 per thousand +/- 2.52 per thousand respectively, with the significant differences (P < 0.05). With generation adding, the portion of S phase increased for mine dust-exposed epithelial cells. (2) At the 26th generations, fibroblasts expressed alpha-SMA. Co-cultured with epithelial cell, the alpha-SMA expression of fibroblasts increased. Especially, positive cell numbers and intensity of staining dramatically increased with generation adding.</p><p><b>CONCLUSIONS</b>(1) The tin mine dust can induce malignant transformation of human pulmonary epithelial cells BEAS-2B and activation of fibroblasts WI-38. (2) The epithelial cells are major target in carcinogenesis induced by Yunnan tin mine dust. (3) Transformation of epithelia and activation of fibroblasts co-evolve in the developing process of induced lung cancer by Yunnan tin mine dust.</p>