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BACKGROUND:Primary culture in vitro of neurons plays an important role in the development, regeneration, signal transduction mechanisms, neuropharmacology and gene expressions of the nervous system. OBJECTIVE:To establish a simple method for primary culture of high-purity cortical neurons in neonatal Sprague-Dawley rats. METHODS:Cortical tissues were acquired from neonatal Sprague-Dawley rats born 1 day. In traditional experimental group, the whole cortex was removed;in improved experimental group, the cortical tissues, 2-3 mm thick on the brain surface were removed. Single cel suspensions were prepared after papain digestion and centrifugation and were then seeded onto 24-wel culture plates containing neuron solutions for primary culture (1×105 per wel ). Cel s were identified by neuronal specific markers MAP-2 and Tuj1 after 3-day culture. The number of neurons and neurite length were observed under inverted phase contrast microscope and recorded at 6, 24, 48 and 72 hours, 5 and 7 days of culture, resprctively. RESULTS AND CONCLUSION:The cultured cel s expressing MAP-2 and Tuj1 were neurons that could be used in the fol owing experiments. The purity of neurons in the improved experimental group was 92%at 3 days, while only 51%in the traditional experimental group. Cel s in both two groups had attached to the wal presenting with smal processes at 6 hours, and a simple neural network formed at the 3rd day until dense neural networks could be found at the 5th day. To conclude, our culture method herein is simple and convenient, and can be used to produce neurons with high purity, which wil be helpful for the experimental studies on cortical neurons from Sprague-Dawley rats.
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Summarize all kinds of methodology of scar’s treatment,reflect the condition of research on the treatment of scar,and outlook the prospect of traditional Chinese medicine on scars in the future.
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Objective To probe the classification of diabetic retinopathy (DR) and the different grade of diabetic and type of macular edema according to fundus fluorescein angiography (FFA). Methods FFA was performed on 1 058 patients (2 097 eyes) to classify DR and macular edema with the analysis of duration of DM, visual acuity, manifestation of FFA images and results of ophthalmoscopic examination. Results In 2 097 eyes, there were 124 (5. 9%) without DR, 396 (18. 9%) with DR I, 430 (20. 5%) with DR II, 563 (26.8%) with DR III, 262 (12. 5%) with preproliferative diabetic retinopathy ( PPDR), 254 (12.%) with DR IV, 60 (2. 9%) with DR V, and 8 (0. 4%) with DR VI. In 2 097 eyes there were 819 (39. 1%) with macular edema, including 311 (38%) with focal macular edema, 322 (39. 3%) with diffused macular edema, 112 (13. 7%) with cystoid macular edema, 25 (3. 1%) with ischemia macular edema, and 49 (6. 0%) with proliferative macular edema. Conclusion With the analysis of the results of FFA of 2 097 eyes, we classify DR in stage Ⅰ (primary stage Ⅰ and Ⅱ), Ⅱ (primary stage Ⅲ), Ⅲ (preproliferative diabetic retinopathy), Ⅳ,Ⅴ, and Ⅵ; classify macular edema of DR in focal, diffused, cystoid, ischemic, and proliferating ones.
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Objective To observe the therapeutic efficacy of endoscopic pancreaticobiliary duct drainage for acute necrotizing pancreatitis. Methods 41 cases of acute necrotizi ng pancreatitis were randomly divided into pancreaticobiliary duct drainage (21 causes) group and control group (20 cases).Results The procedure was successful in all 21 cases. The difference bet ween the two groups was statistically significant including hospital stay 〔(2 8?12) days vs. (37?19) days,P
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Objective To explore the expression and change on the neurokinin A(NKA) of rat ileum during the development. Methods PAP immunohistochemistry method and image analysis were used to detect systematically the expression of NKA in rat ileum from E13 to adult stage. Results 1.In rat ileum, the NKA-IR could first be found in the myenteric plexus at E17, and then appeared on the longitudinal muscle layer, circular muscle layer, intestinal villus, intestinal gland, submucosa and deep muscular plexus. On P30, they showed the distribution feature of adult rat.2.The results of quantitative analysis were in correspondence with the developmental change of NKA-IR in ileum.3.The NKA-IR positive cells showed the typical feature of morphology and distribution of gastrointestinal endocrine cells.Conclusion 1. The ontogeny and development of NKA in rat ileum mainly appeare in the last E1 week and P4 weeks, whereas the density of positive fiber in villus is not as same as adulthood until P60. 2.There are two key periods during the development of NKA in rat ileum. The first crucial stage is from the last El week to P1 week. The second important stage is in the end of P4 weeks. 3.The NKA-IR cells may be the endocrine cell which produces NKA in rat digestive tract.4.NKA may play an important role in the development and physiological function of rat ileum.