ABSTRACT
Objective To establish a low density, high purity and high stability in vitro culture method of primary hippocampal neurons of fetal rats by co-culturing hippocampal and cortical cells, so as to obtain higher purity and better vitality of primary hippocampal neurons disease. Methods The fetal rat hippocampal tissue was isolated from 16-18 days pregnant SD rats, then cut and digested by 0.125% trypsin. The obtained cell suspension was filtered by 200 mesh cell sieve, and then the obtained cell suspensions were then inoculated into the inner layer and outer ring of the culture plate in a surrounding form. They were co-cultured in DMEM/F12 medium containing 10% horse serum. After 4-6 hours of cell adhesion, the culture medium was changed to maintenance medium (Neurobasal+2% B27+0.5 mmol/L glutamine). Then the cell viability was detected with CCK-8 kit and the purity of hippocampal neurons was detected by immunofluorescent staining. Results Hippocampal neurons grew well and formed crisscross neural networks after 5 days. And it could survive for 3 weeks. The purity of hippocampal neurons was up to 98%. Conclusion The method of co-culturing hippocampal and cortical cells can obtain high-purity, high activity, high survival rate, and high stability primary hippocampal neurons from fetal rats, which can provide certain experimental conditions for the study of hippocampal neuron related diseases in the nervous system and is worthy of promotion and application.
ABSTRACT
Huangdi Neijing (Yellow Emperor 's Canon of Medicine) establishes the theory that yuan-source points are indicated in zangfu diseases. However, compared with the yuan-source points of yin meridians for the treatment of zang-organ diseases, there is less attention to the indication of yuan-source points of yang meridians for fu-organ diseases, and this statement is even questioned. Collating the early literature and associating with the researches of medical experts, it is found that Nanjing (Classic of Difficult Questions) should be the theoretic origin that yuan-source points of yang meridians are indicated in fu-organ diseases. Regarding the reasons why this theory has not received clinical attention, the three aspects are involved, i.e. the theoretic completion of "he-sea points of three-foot-yang meridians for the diseases of six fu-organs", the limitation of the theory itself, and the lack of literature materials. It is proposed that the exploration on this theory should be deepened in views of the essence of yuan-source points relevant with the characteristic of the wrist-ankle pulse palpation region, acupoint combination, modern technology, etc.
Subject(s)
Meridians , Acupuncture Points , Foot , Lower Extremity , MedicineABSTRACT
<p><b>OBJECTIVE</b>To explore the therapeutic effectiveness of Ilizarov technique in treatment of infected tibial defects combined with overlaying skin defects.</p><p><b>METHODS</b>Twenty-one cases with infected tibial defects combined with skin defects were treated between 2001 and 2008 includeing 18 males and 3 females with an average age of 31 years ranging from 19 to 43 years. The length of bone defect ranged from 3 to 13 cm (means 6 cm). Skin defect area was from 3 cm x 3 cm to 6 cm x 10 cm; 11 cases combined with drop foot, 5 cases with arthrocleisis of knee. Preoperative X-ray of the affected limb was performed and zone of skin necrosis was marked, then the point and length of osteotomized bone, and scope of bone and soft tissue need for removing were determined. The internal fixation were removed. Opening irrigation, vacuum sealing drainage (VSD), and dressing changing were appllied. The skin was fixed with Kirschner wire and bone was transferred with Ilizarov technique in all patients. The lengthening of bone and skin was carried out for 4 to 7 days after surgery, 1/6 to 1/4 mm once, 4 to 6 times a day. The clinical effectiveness was determined mainly through wound and lengthening of skin.</p><p><b>RESULTS</b>All patients were followed up for from 6 to 62 months (means 49.5 months). Fourteen of 21 cases received one stage treatment, there was still secretion from end of bone in 3 patients whose bone healed after debridement, the other 4 patients were cured via trimming end of bone and compression fusion. The defects of bone were extended to full length in 18 patients. Abutting end was slightly absorbed and became rattailed in 2 cases, there was lack of blood supply to abutting ends in one patient who was cured via bone graft from iliac bone. Skin defects was cured in 18 patients with one stage treatment, the other 3 patients were cured after infection was controlled. The deformity of drop foot were corrected in 11 patients, and function of knee was improved in five patients. The external fixator was removed at 1.2 to 2.6 years after surgery. At last, bone infections were cured, defects of bone and skin recovered in all patients.</p><p><b>CONCLUSION</b>One stage treatment of infected tibial defects combined with skin defects using Ilizarov technique has minimal invasion with less complex surgeries, could reduce the time and expense of treatment.</p>