1.Optimization of culture conditions for oligodendrocytes of the rat cerebral cortex
Kai YANG ; Yipeng LI ; Yingfu LIU ; Yuanchi CHENG ; Fengwu TANG ; Bing LIANG ; Zhongwei XU ; Xuyi CHEN
Chinese Journal of Tissue Engineering Research 2016;20(29):4328-4333
BACKGROUND:Oligodendrocytes are mostly differentiated from oligodendrocyte precursor cel s. A suitable medium and cel seeding density have a significant impact on the process of the isolation of oligodendrocyte precursor cel s to obtain oligodendrocytes. OBJECTIVE:To explore the optimization of oligodendrocyte culture conditions. METHODS:Oligodendrocyte precursor cel s isolated from the newborn rats 48 hours after birth were cultured in DMEM/high glucose medium or DMEM/F12 medium using seeding densities of 2×104 cel s/cm2, 4×104 cel s/cm2, 8×104 cel s/cm2, 16×104 cel s/cm2, 32×104 cel s/cm2, and 64×104 cel s/cm2, respectively. Oligodendrocyte precursor cel s were induced to differentiate into oligodendrocytes at 72 hours after cel adhesion. Morphology of differentiated oligodendrocyte precursor cel s were observed under a light microscope, and the differentiation results were identified by immunofluorescence staining after 7-day induced differentiation. RESULTS AND CONCLUSION:Morphology of oligodendrocyte precursor cel s were recognized when cultured in DMEM/high glucose medium or DMEM/F12 medium using seeding densities of 2×104 cel s/cm2, 4×104 cel s/cm2, and 8×104 cel s/cm2, respectively. Immunofluorescence staining showed that myelin basic protein-positive cel s were found after 7-day induced differentiation, and the positive cel number were 16.40±3.30, 49.95±2.33, and 76.95±4.86 in DMEM/F12 medium, and 12.65±2.53, 32.10±1.17, and 54.05±1.56 in DMEM/high glucose medium (P<0.05). These findings indicate that DMEM/F12 medium is more suitable for culturing oligodendrocyte precursor cel s compared with DMEM/high glucose medium to some extent. The number of differentiated oligodendrocytes was gradual y increased with the enhanced seeding density of oligodendrocyte precursor cel s, and the seeding densities from 4×104 to 8×104 cel s/cm2 were appropriate for the observation of cel morphology.
2.Early diagnosis of acute bilirubin encephalopathy in the newborn
Danhua MENG ; Xinnian PAN ; Dan ZHAO ; Yan LI ; Qiufen WEI ; Hongjuan BI ; Kaiyan SHEN ; Jing XU ; Jinmei GAN ; Yingfu LIANG
Chinese Journal of Neonatology 2017;32(5):346-350
Objective To investigate the diagnostic correlation and sensitivity of amplitude integrated electroencephalogram (aEEG),brainstem auditory evoked potential (BAEP) and cranial magnetic resonance imaging (MRI) for acute bilirubin encephalopathy (ABE) in the newborn.Method Term and near-term neonates (gestational age ≥ 35 weeks) with hyperbilirubinemia (the level of bilirubin over than 95th percentile) of high and intermediate risk group admitted in the neonatal ward of Guangxi Maternal and Child Health Care Hospital from Jan 2014 to Dec 2015 were recruited retrospectively.The infants were assigned to ABE group and non-ABE group according to the diagnostic criteria of ABE.The clinical data of the newborns were collected and the diagnostic correlation between clinical diagnosis and aEEG,BAEP and cranial MRI were analyzed.The receiver operating characteristic (ROC) curve was adopted to assess the diagnostic efficiency of the peak level of serum bilirubin,aEEG,BAEP and cranial MRI on the early diagnosis of ABE.Result A total of 152 newborns with hyperbilirubinemia were recruited,including 33 cases in the ABE group and 119 cases in non-ABE group.(1) The results of aEEG and MRI were marginally positively correlated with clinical diagnosis of ABE (aEEG:r =0.487,P < 0.001;MRI:r =0.220,P=0.018),while the results of BAEP were closely related to the clinical diagnosis of ABE (r =0.593,P < 0.001);(2) The results of BAEP and MRI on the diagnosis of ABE were positively correlated with those of aEEG (BAEP:r =0.424,P < 0.001;MRI:r =0.307,P < 0.001).(3) The area under the ROC curves for predicting the onset of ABE were 0.899 for the peak level of serum bilirubin,0.767 for BAEP,0.738 for aEEG and 0.590 for MRI.Conclusion There was the correlation on the diagnosis of ABE among the methods of aEEG,BAEP and MRI.The combined diagnosis of the three methods could play a complementary role.The aEEG contributed to the early diagnosis of ABE with high sensitivity.
3.Analysis of Composition Principles of Prescriptions for Simple Obesity Based on Traditional Chinese Medicine Inheritance Support System
Huiyan ZENG ; Yingfu XIAO ; Jinming ZHANG ; Xianyu TANG ; Liu HE ; Qiting LIANG ; Jiali HE ; Lu SUN ; Wenwen XIE
Journal of Guangzhou University of Traditional Chinese Medicine 2017;34(4):612-616
Objective To explore the composition principles of prescriptions for simple obesity based on the analysis of medical literature issued in recently 30 years about Chinese medicine for simple obesity. Methods We collected literature of Chinese medicine in treating simple obesity from VIP, CNKI, Wanfang, CBM and CMCC/CMCI databases, and screened out the formulas for simple obesity to establish a formula database. And then the medication rules were figured out with the Traditional Chinese medicine Inheritance Support System version 2.5 (TCMISS 2.5) software. Results A total of 57 formulas were screened out. And the top 10 herbs with highest occurrence frequency in the prescriptions were Poria, Rhizoma Atractylodis Macrocephalae, Rhizoma Alismatis, Radix et Rhizoma Rhei, Fructus Crataegi, Radix Astragali, Radix Glycyrrhizae, Folium Nelumbinis, Pericarpium Citri Reticulatae and Semen Cassiae. After data mining, we got 13 high-frequency herb-pair combinations and 31 core combinations, and 8 new prescriptions were extracted. Conclusion The research has achieved the quantitative description of the relationship between the Chinese medicines, the extraction of the core combinations and the discovery of new Chinese medicine prescriptions for simple obesity. The results will provide references for the development of new Chinese medicines for the treatment of simple obesity .