BACKGROUND:Bone marrow mesenchymal stem cells(MSCs)are a kind of adult stem cells with multi-potential differentiation property.At present,it has served as cell carrier for the treatment of Parkinson's disease.OBJECTIVE:To construct pDsRed-C1-CDNF eukaryotic expression vector and induce its expression in rat MSCs.METHODS:CDNF gene was amplified from mouse tissues using RT-PCR,and sequence with Xho I,BamHI restriction enzyme cutting site.The CDNF gene was inserted into the eukaryotic expression vector pDsRed-C1 encoding red fluorescent protein gene.The plasmid pDsRed-C1-CDNF was constructed and transfected into rat bone marrow MSCs.RESULES AND CONCLUSION:The pDsRed-C1-CDNF recombinant plasmid was confirmed by double digestion of Xho I and BamHI restriction enzyme or single digestion of BamHI,and PCR sequence.Results show that the recombinant pDsRed-C1-CDNF eukaryotic expression vector was successfully constructed.

Objective To explore the efficacy and the prognostic factors of docetaxel combined with gemcitabine for patients with recurrent or metastatic breast cancer.Methods Forty-six patients with recurrent or metastatic breast cancer received docetaxel combined with gemcitabine regimen (docetaxel 75 mg/m2,intravenous drip,dl ; gemcitabine 1000 mg/m2,intravenous drip,dl,d8; The regimen was repeated every threeweeks for 3-6 cycles).The response rate (RR) was evaluated after 2 cycles,and the overall survival (OS)and progress-free survival (PFS) were recorded.Single factor chi square test and multivariate Cox proportion hazard model were used to evaluate the relationship between clinic pathologic features and RR,OS.Results All patients completed 3-6 cycls chemotherapy.The overall response rate was 56.5％ (26/46) and disease control rate was 82.6％ (38/46),with 4 patients of complete remission (8.7％),22 patients of partial remission (47.8％),12 patients of stable disease (26.1％) and 8 patients of progressive disease (17.4％).The median OS of 46 patients was 16.0 months (95％ CI:6.5-25.5 months) and PFS was 8.0 months (95％ CI:6.2-9.8 months).Single factor analysis showed that age,menopausal status,PS score,the number of metastasis had correhtions with RR (P ＜ 0.05) ; Cox proportion hazard model also showed that age,menopausal status,PS score,the number of metastasis had correlations with OS,and were the prognostic factors (P ＜0.05).The most frequent treatment-related adverse events were myelosuppression,gastrointestinal reaction,rash,alopecia,fatigue,and were tolerable (Ⅰ-Ⅱ level).Condusion Docetaxel combined with gemcitabine is effective and safe in the treatment of recurrent or metastatic breast cancer.Age,menopausal status,PS score and the number of metastasis are the prognostic factors for efficacy of docetaxel combined with gemcitabine regimen.

Mutations in genes encoding amyloid precursor protein (APP) and presenilins (PSs) cause familial forms of Alzheimer's disease (AD), a neurodegenerative disorder strongly associated with aging. It is currently unknown whether and how AD risks affect early brain development, and to what extent subtle synaptic pathology may occur prior to overt hallmark AD pathology. Transgenic mutant APP/PS1 over-expression mouse lines are key tools for studying the molecular mechanisms of AD pathogenesis. Among these lines, the 5XFAD mice rapidly develop key features of AD pathology and have proven utility in studying amyloid plaque formation and amyloid β (Aβ)-induced neurodegeneration. We reasoned that transgenic mutant APP/PS1 over-expression in 5XFAD mice may lead to neurodevelopmental defects in early cortical neurons, and performed detailed synaptic physiological characterization of layer 5 (L5) neurons from the prefrontal cortex (PFC) of 5XFAD and wild-type littermate controls. L5 PFC neurons from 5XFAD mice show early APP/Aβ immunolabeling. Whole-cell patch-clamp recording at an early post-weaning age (P22-30) revealed functional impairments; although 5XFAD PFC-L5 neurons exhibited similar membrane properties, they were intrinsically less excitable. In addition, these neurons received smaller amplitude and frequency of miniature excitatory synaptic inputs. These functional disturbances were further corroborated by decreased dendritic spine density and spine head volumes that indicated impaired synapse maturation. Slice biotinylation followed by Western blot analysis of PFC-L5 tissue revealed that 5XFAD mice showed reduced synaptic AMPA receptor subunit GluA1 and decreased synaptic NMDA receptor subunit GluN2A. Consistent with this, patch-clamp recording of the evoked L23>L5 synaptic responses revealed a reduced AMPA/NMDA receptor current ratio, and an increased level of AMPAR-lacking silent synapses. These results suggest that transgenic mutant forms of APP/PS1 overexpression in 5XFAD mice leads to early developmental defects of cortical circuits, which could contribute to the age-dependent synaptic pathology and neurodegeneration later in life.
Mice ; Animals ; Alzheimer Disease/pathology* ; Amyloid beta-Peptides/metabolism* ; Receptors, N-Methyl-D-Aspartate/metabolism* ; Amyloid beta-Protein Precursor/metabolism* ; Mice, Transgenic ; Neurons/metabolism* ; Receptors, AMPA/metabolism* ; Disease Models, Animal