Primary culture and identification of rat glomerular microvascular endothelial cells / 生理学报

The aim of the present study was to establish a simple and efficient method for isolation and culture of primary rat glomerular microvascular endothelial cells in vitro. The bilateral kidneys were taken from 7-10-day old Sprague-Dawley rats, and the renal cortex was separated. Glomeruli were obtained by cutting and continuously passing 200-mesh and 300-mesh sieves. After type IV collagenase digestion for 15-20 min, renal microvascular globules were collected for inoculation and culture. The cultured cells were identified by cell morphology observation and immunocytochemical staining with factor VIII related antigen. The results showed that the renal microvascular globules were irregularly spherical, without cysts, and the capillary loop structure was clear; after 3 days of primary culture, short spindle-shaped cells crawled out around the renal microvascular globules and gradually formed cell colonies, showing an "island-like shape" distribution; 4-5 days later, the cell colonies fused with each other; 6 days later, the cells covered the bottom of the dish, showing a typical monolayer, paving stone-like, mosaic arrangement. The immunocytochemical staining of factor VIII related antigen showed that the cytoplasm was lightly stained brownish red, and factor VIII related antigen-positive rate of cells was nearly 100%. The above results suggested that this study successfully established a method combining continuous screening and collagenase digestion for culture of primary rat glomerular microvascular endothelial cells in vitro. It provides an important tool cell for studying the mechanism of the occurrence and development of diabetic nephropathy.

Long term follow up of midline partial glossectomy and uvulopalatopharyngoplasty for treating obstructive sleep apnea hypopnea syndrome / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To observe the long-term results of midline partial glossectomy with uvulopalatopharyngoplasty (UPPP) in the treatment of obstructive sleep apnea hypopnea syndrome (OSAHS).</p><p><b>METHODS</b>Twenty-four severe OSAHS patients treated with midline partial glossectomy and UPPP from January 2003 to March 2004 were included in this study, the follow-up was 5 years. The median of preoperative lowest arterial oxygen saturation (LSaO(2)) of this group at night (the same below) 0.650, and AHI was 56.5 times/h, UPPP was performed under general anesthesia, no tracheotomy performed. Criteria of curative effects: AHI < 5 times/h was recovery, AHI < 20 times/h and decreased beyond 50% marked improvement, only AHI decreased beyond 50% improvement.</p><p><b>RESULTS</b>Post-operation AHI (6 months, 1 year, 2 years and 5 years after surgery) decreased significantly compared to that before the surgery, and post-operation LSaO(2) was significantly higher than that of preoperative (Wilcoxon's signed rank test, the same below, P < 0.01). The LSaO(2) and AHI were significantly different between 1 year, 2 years, 5 years and 6 months post-operatively (P < 0.01). Six months after surgery, PSG results showed that 21 were recovery, marked improvement for the other 3 cases, the recovery rate was 87.5%. One year after surgery, 18 were recovery, marked improvement in 3 cases, the recovery rate 75.0%. Two years after surgery, 14 cases recovery, marked improvement in 4 cases, the recovery rate 58.3%. Five years after surgery, 6 were recovery, the recovery rate 25.0%. Among 5 cases with hypertension before the surgery, after surgery antihypertensive drugs were not necessary in 4 cases, and the dosage was decreased in 1 case.</p><p><b>CONCLUSION</b>The midline partial glossectomy with UPPP surgery may be an effective treatment for the severe OSAHS, long-term effect is satisfactory.</p>