BACKGROUND:PA6 cells are bone marrow stromal stem cells from the mouse skul , and scientists have found that PA6 cells co-cultured with some kinds of stem cells have shown neural differentiation, based on which, PA6 cells can be used for repair of nerve injury. Therefore, researchers give more and more attentions to PA6, but few studies have addressed isolation and culture methods of bone marrow stromal stem cells from the skul . OBJECTIVE:To isolate and culture bone marrow stromal stem cells from the skul of Sprague-Dawley rats and to observe cellular morpholopy in vitro and perform immunofluorescence identification. METHODS:Under sterile conditions, newborn Sprague-Dawley rat’s skul was cut into pieces. Smal skul pieces were washed using Dulbecco’s modified Eagle’s medium containing 10%fetal bovine serum. Single cellsuspension was made, and placed in culture flask. The culture medium was changed many times for cellpurification. The second passage of cells was obtained for morphology observation under an inverted microscope. cellsurface markers were detected by using immunofluorescence staining. RESULTS AND CONCLUSION:After the primary culture for 24 hours, cells exhibited adherent growth;after 3 days, cells were increased in number, and presented with irregular shapes, such as polygon, triangle, spindle and flat shape. After passage, cells were uniform in morphology, arranged in radial and bunch patterns, and exhibited strong adherent ability. Some cells grew in cluster, and proliferated faster than primary cells. Immunofluorescence staining showed that cells were positive for CD105, CD73, CD44, CD90, but negative for CD45, D34,CD14, HLA-DR. Results indicate that this method can obtain bone marrow stromal stem cells.

Objective Based on the Traditional Chinese Medicine Inheritance Support System (TCMISS) (V2.0), to analyze the point-selection rules in acupuncture-moxibustion prescriptions for gastroptosis indexed by China National Knowledge Infrastructure (CNKI), Wanfang and Vip databases, and to obtain novel prescriptions, for providing clinical references. Method Acupuncture-moxibustion prescriptions for gastroptosis indexed by CNKI, Wanfang and Vip databases were collected and filtered, and then input into the TCMISS. The prescriptions were analyzed by using data mining method. Result The frequency and core combination of the commonly-used acupoints were determined out of the 76 eligible prescriptions for gastroptosis, and 3 novel prescriptions were obtained. Conclusion The TCMISS is an important tool in mining and analyzing the point-selection rules in acupuncture-moxibustion treatment, and the new acupoint prescriptions generated by this system provide references to the treatment of gastroptosis.