Directed differentiation of human induced pluripotent stem cells into midbrain / 南方医科大学学报

OBJECTIVE@#To establish an efficient protocol for directed differentiation of human induced pluripotent stem cells (hiPSCs) into functional midbrain dopaminergic progenitor cells (DAPs) in vitro.@*METHODS@#hiPSCs were induced to differentiate into DAPs in two developmental stages. In the first stage (the first 13 days), hiPSCs were induced into intermediate cells morphologically similar to primitive neuroepithelial cells (NECs) in neural induction medium containing a combination of small molecule compounds. In the second stage, the intermediate cells were further induced in neural differentiation medium until day 28 to obtain DAPs. After CM-DiI staining, the induced DAPs were stereotactically transplanted into the right medial forebrain bundle (MFB) of rat models of Parkinson's disease (PD). Eight weeks after transplantation, the motor behaviors of PD rats was evaluated. Immunofluorescence assay of brain sections of the rats was performed at 2 weeks after transplantation to observe the survival, migration and differentiation of the transplanted cells in the host brain microenvironment.@*RESULTS@#hiPSCs passaged stably on Matrigel showed a normal diploid karyotype, expressed the pluripotency markers OCT4, SOX2, and Nanog, and were positive for alkaline phosphatase. The primitive neuroepithelial cells obtained on day 13 formed dense cell colonies in the form of neural rosettes and expressed the neuroepithelial markers (SOX2, Nestin, and PAX6, 91.3%-92.8%). The DAPs on day 28 highly expressed the specific markers (TH, FOXA2, LMX1A and NURR1, 93.3-96.7%). In rat models of PD, the hiPSCs-DAPs survived and differentiated into TH+, FOXA2+ and Tuj1+ neurons at 2 weeks after transplantation. Eight weeks after transplantation, the motor function of PD rats was significantly improved as shown by water maze test (P < 0.0001) and apomorphine-induced rotation test (P < 0.0001) compared with rats receiving vehicle injection.@*CONCLUSION@#HiPSCs can be effectively induced to differentiate into DAPs capable of differentiating into functional neurons both in vivo and in vitro. In rat models of PD, the transplanted hiPSCs-DAPs can survive for more than 8 weeks in the MFB and differentiate into multiple functional neurocytes to ameliorate neurological deficits of the rats, suggesting the potential value of hiPSCs-DAPs transplantation for treatment of neurological diseases.

A new isobenzofuranone derivative from Phlomis betonicoides and its antibacterial activity / 中国中药杂志

A new isobenzofuranone derivative has been isolated from Phlomis betonicoides by using various chromatographic techniques, including silica gel, Sephadex LH-20, MCI-gel resin and RP-HPLC. This compound was determined as 5-(3-hydroxypropyl)-2,2-dimethyl-2-furo[3,4-]chromen-7(9)-one (1) by NMR, MS, IR and UV spectroscopic data. Compound 1 showed potent antibacterial activity with an MIC₉₀ value of (58.4 ± 4.2) mg·L⁻¹ for methicillin resistant Staphylococcus aureus (MRSA) strain [levofloxacin as a control with MIC₉₀ value of (52.8±4.6) mg·L⁻¹].

Q-analysis real-time ultrasonic elastography quantitative analysis in the diagnosis of breast benign and malignant lesions / 中华医学超声杂志（电子版）

Objective To evaluate the value of GE′s Q-analysis real-time ultrasonic elastography quantitative analysis software in the diagnosis of breast diseases and explore the best cutoff point of the breast diseases diagnosis.Methods The elastograms of 71 breast lesions(66 patients)were evaluated by using the GE LOGIQ E9 with the Q-analysis software.All the 71 lesions were diagnosed as malignant and benign lesions by surgery or biopsy.The receiver operating characteristic (ROC)curves of the entirety elasticity rate and local elasticity rate was performed.We compared the areas under ROC curves ( AUC) of the two modalities and confirmed the best cutoff point of the breast diseases diagnosis .We also assessed the sensitivity and specificity of the two modalities .Lastly we assessed the ROC curves with Z test to determine whether the difference between the two modalities was significant .Results Among the 71 leisions (66 patients),there were 37 benign lesions and 34 malignant ones.The elastograms of the leisions showed that the elasticity of 37 benign leisions was good ,the hardness value was low and the texture was even .As for the 34 malignant leisions,the elasticity was bad,the hardness value was high and the texture was uneven .The AUC of the local modality was 0.899 ( >0.5).The AUC of the entire modality,was 0.985 ( >0.5).Both modalities were accurate for the diagnosis of breast diseases .When the cutoff point of entirety elasticity rate was set as 3.60,the sensitiveness was 82.4%and the specificity was 86.5%.When the cutoff point of local elasticity rate was set as 2.60,the sensitiveness was 97.1% and the specificity was 91.9%.The difference between the two modalities was significant (Z =2.621,P=0.0088).Conclusions Q-analysis real-time ultrasonic elastography quantitative analysis is able to evaluate the rigidity and texture of tissue and is valuable to distinguish benign breast lesions between malignant breast lesions .The sensitiveness and specificity of local modality are higher than that of entirety modality .