BACKGROUND:Compared with the bone marrow mesenchymal stem cells (BMSCs),cells from the umbilical cord blood (UCB)are considered"very young",and its proliferation and differentiation cannot decrease with age.Thus,cells from UCB are a great substitute of BMSCs.OBJECTIVE:To observe the characteristics of in vitro differentiation of UCB-MSCs into cardiomyocytes.DESIGN,TIME AND SETTING:The observational experiment was performed at the Beijing Sijitan Hospital from March 2005 to February 2007.MATERIALS:Cells from UCB were collected from healthy arturients after signing the informed consent.METHODS:Mononuclear cells were isolated to further harvest MSCs.At the third passage,CD34,CD44 and CD90 were measured using immunofluorescence flow cytometry.After 4 weeks of 5-azacitidine treatment,myosin heavy chain,GATA4 and troponin 1 were identified using immunohistochemistry and reverse transcription-polymerase chain reaction.MAIN OUTCOME MEASURES:β-myosin heavy chain,GATA4 and troponin I expression.RESULTS:UCB-MSCs showed a fibrolast-like morphology,clonally expanded after 5-azacitidine reatment.The immunophenotype of these clonally expanded cells is consistent with that reported for BMSCs.Immunohistochemistry and reverse transcription-polymerase chain reaction showed β-myosin heavy chain,GATA4 and troponin I expression.CONCLUSION:UCB-MSCs differentiate into cardiomyocyte-like cells,which have the characteristics of cardiomyocytes.

BACKGROUND: Umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) have been shown to lead to new tissue formation after homing and engrafting to the heart. But the safety of UCB-MSCs engrafting remains to be further investigated. OBJECTIVE: To study the safety and apoptosis inhibition of the UCB-MSCs under co-culture conditions on human cardiomyocytes. METHODS: UCB was collected at delivery with informed consent obtained from 10 donors. The UCB-MSCs were treated with 5-azaserine to induce differentiation into cardiomyocytes. The in vitro cultured cells of the 3rd-5~(th) passages and dividing cells were taken to detect telomerase activity, tumor-related gene expression, G-banding patterns of chromosomal karyotupes, cell surface antigen expression, tumor formation in nude mice, and inhibited apoptosis under co-culture conditions. RESULTS AND CONCLUSION: Prior to and after 5-azaserine induction, telomerase activity and tumor-related gene expression (p53, cyclin A, cdk2, β-actin, C-fos, h-TERT, c-myc) of UCB-MSCs were similar, no abnormal chromosomal karyotupes were observed, immunophenotype exhibited no change, CD34 was negative, but CD44 and CD90 (Thy-1) were positive. At 10 weeks after inoculation of UCB-MSCs, nude mice still survived healthily and no formed tumor in vivo was observed. Hematoxylin-eosin staining suggested normal subcutaneous tissue. Compared with simple cardiomyocytes, UCB-MSCs could significantly inhibit cardiomyocyte apoptosis under co-culture conditions (P < 0.05), indicating that human UCB-MSCs are a valuable, safe, and effective source of cell transplantation treatment.

Objective To study the safety and effect of the umbilical cord blood(UCB)-derived mesenchymal stem cells(MSCs)on apoptosis of human cardiomyocytes(HCM). MethodsUCB was collected at the time of delivery with informed consent obtained from 10donors.The UCB-derived MSCs were treated with 5-azaserube(5-AZA)and were further induced to differentiate into cardiomyocytes.Telomerase activity,G-banding patterns of chromosomal karyotypes,tumor formation in nude mice,RT-PCR,and the effect of inhibiting apoptosis of HCM were investigated. ResultsMSCs derived from UCB were differentiated into cardiomyocytes in vitro,which possessed telomerase activity after 5-AZA induction,and no abnormal chromosomal karyotypes were observed.Expression of p53,cyclin A,cdk2,β-actin,C-fos,h-TERT and c-myc were similar in MSCs before and after 5-AZA treatment.There was no tumor formation in nude mice after injection of UCB-derived MSCs.UCB-derived MSCs significantly inhibited apoptosis of HCM. ConclusionUCB-derived MSCs are a valuable,safe and effective source of cell-transplantation treatment.

Aphasia is one of the major complications after stroke, which needs an effective assessment. Event-related potential (ERP) has been widely researched in neurology, the endogenous components, such as P300, N400, mismatch negativity (MMN), contingent nega-tive variation (CNV) have been widely used in diagnosis and evaluation of the impairment of brain function, including the language func-tion. This paper discussed the application of different endogenous components of ERP in aphasia after stroke, especially the comparative analysis of N400 and P300.

The relevant policies of national essential medicine system were reviewed, major concerns and actions in the implementation of essential medicine system in Shanghai were introduced, and the suggestions to improve the implementation of essential medicine system of Shanghai were made. These provided the information for policy making and provided a useful experience for facilitating the establishment of essential medicine system and the improvement of its implementation in Shanghai as well as China.

Objective To investigate the performance of MicroScan WalkAway 96 Plus (MSW) system in detection of carbapenem-resistant Enterobacteriaceae (CRE).Methods A total of 81 stock CRE strains were used in this study. Bacterial identification and antimicrobial susceptibility test were performed by MSW system. Beta-lactamases genes blaKPC,blaIMP,blaVIM, blaOXA-48 and blaNDM were amplified by PCR and subjected to sequencing analysis. Disk diffusion method and PCR were used as gold standard to evaluate the performance and reliability of MSW system in identifying carbapenem-resistant and carbapenemase-producing Enterobacteriaceae.Results Overall, 69.1 % (56/81) of the Enterobacteriaceae strains were identified as CRE by the MSW system. The results of PCR showed that 48 strains were carbapenemase-producing Enterobacteriaceae. When carbapenemase-producing Enterobacteriaceae strains were identified by the instrument using an advanced expert system, the sensitivity was 93.8 % and specificity was 42.4 %. The positive predictive value was 70.3 %, the negative predictive value was 82.4 % and the predictive accuracy value was 72.8 %.Conclusions The MicroScan WalkAway 96 Plus system has shown good performance in detection of CRE.

OBJECTIVE:To establish a method for the simultaneous determination of 5 unsaturated fatty acids in Perilla oil cap-sule. METHODS:With the reference material of α-linolenic acid methyl ester,GC was used to determine and calculate the relative correction factors of α-linolenic acid methyl ester with methyl palmitate,methyl stearate,methyl oleate and linoleic acid methyl es-ter,and the correction factors were used to calculate the contents of 5 unsaturated fatty acids;the column was Agilent Innowax cap-illary column,the detector was FID,the inlet temperature was 230 ℃,the detector temperature was 250 ℃,the gas flow rate was 20 ml/min(nitrogen),40 ml/min(hydrogen)and 350 ml/min(air),split ratio was 30 to 1,the column temperature was 190 ℃, and injection volume was 1 μl. RESULTS:The linear range was 0.018-0.792 μg(r=0.9994)for methyl palmitate,0.0016-0.0176μg(r=0.9993)for methyl stearate,0.0056-0.2464 μg(r=0.9999)for methyl oleate,0.003-0.132 μg(r=0.9990)for linoleic acid methyl ester and 0.018-0.792 μg(r=0.9998) for α-linolenic acid methyl ester;RSDs of precision,stability and reproducibility tests were lower than 5%;recoveries were 98.990%-101.70%(RSD=0.720%,n=6) for methyl palmitate,99.599%-100.699%(RSD=0.368%,n=6) for methyl stearate,98.996%-101.680%(RSD=1.240%,n=6) for methyl oleate,99.813%-100.963%(RSD=0.434%,n=6)for linoleic acid methyl ester and 97.185%-99.602%(RSD=0.874%,n=6)for α-linolenic acid methyl es-ter. CONCLUSIONS:The method is simple and stable with good reproducibility,and can be used for the simultaneous determina-tion of methyl palmitate,methyl stearate,methyl oleate,linoleic acid methyl ester,α-linolenic acid methyl ester in Perilla oil cap-sule.

ObjectiveTo investigate the effect of macrophages (MCs) on the differentiation of mouse induced pluripotent stem cells (iPSCs) into hepatic progenitor cells (HPCs). MethodsA total of 24 C57BL/6N mice were used to obtain MCs by peritoneal irrigation, and the supernatant was collected to obtain the conditioned medium of MCs (MC-CDM). Activin A, bone morphogenetic protein 4, and fibroblast growth factor were used to induce the differentiation of mouse iPSCs into HPCs. The differentiation of HPCs were randomly divided into control group (normal medium) and experimental group (MC group; use of MC-CDM medium on day 5 of induction). Morphology, immunofluorescence assay, and Western blot were used to compare the morphology of HPCs and the expression of related proteins between the control group and the MC group. The t-test was used for comparison of continuous data between two groups. ResultsHPCs derived from iPSCs were established in vitro, and HPCs had the potential to differentiate into hepatocytes. Immunofluorescence assay showed that compared with the D12 control group, the D12 MC group had a significant increase in the protein expression of the HPC-specific protein CK19 (0.901±0.072 vs 0.686±0.097, t=-3.093, P＜0.05). Western blot showed that compared with the D12 control group, the D12 MC group had a significant increase in the protein expression of the HPC-related protein CK19 (1.922±0.103 vs 1.448±0.012, t =-7.881, P ＜005), as well as a significant increase in the protein expression of the autophagy-related protein LC3 (1.392±0.042 vs 1.101±0048, t =-5.978, P＜005). ConclusionMCs can promote the differentiation of mouse iPSCs into HPCs, possibly by increasing the autophagy level of HPCs.

Objective Ultraviolet radiation can induce the expression of matrix metalloproteinases (MMPs) in pterygium epithelial cells. To investigate the effects of curcumin on the expression of MMP 2 and MMP 9 in human pterygium fibroblasts (HPFs) on UVA-irradiation and its possible mechanism.Methods After optimizing the dose of UVA irradiation and the concentration of curcumin, HPFs in the second generation were divided into control group (no exposure to UVA, no medication), UVA group (exposure to UVA), and UVA + curcumin group (exposure to UVA + curcumin). MMP-2 and MMP-9 levels were tested by zymography. The mRNA expression of MMP-2 and MMP-9 was detected by RT-PCR. The NF-κB-DNA binding activity was detected by electrophoretic mobility shift assay (EMSA).Results Compared with the control group, the secretion of MMP-2 and MMP-9 in the UVA group was increased significantly (P<0.05), which could be suppressed by curcumin (P<0.05). The MMP-9 mRNA levels were also significantly increased in UVA group \[(100±0)% vs (247.0±10.8)%, P<0.05\], and could be inhibited by curcumin \[(88.7±5.1)% vs (247.0±10.8)%, P<0.05\]. The NF-κB-DNA binding was remarkably increased in UVA group (P<0.05), and significantly decreased after treatment with curcumin (P<0.05).Conclusion Curcumin can inhibit the expression of MMP-2 and MMP-9 in UVA-irradiated HPFs, and the mechanism involved in this protective effect might be its inhibitory effect on NF-κB activation.

Precision medicine has become a new mode of modern medicine, and personalized medication is the important embodiment of clinical application of precision medicine. The advances of life science technologies greatly facilitated precision medicine, and also promoted the shift of the mode of clinical pharmacy care from rational drug use and individualized medication to precision medication. To achieve"one person, one mode" clinical dosage regimens,it is necessary to rely on the supports of advanced life science technologies and precisely analyzing and accurately characterizing the biomarker clusters related to individual differences among patients, pathological differences of disease, and disease progression. This article illustrated the recent advances in the application of pharmacokinetics/pharmacodynamics, omics technology and liquid biopsy to the design of dosage regimen, prediction of therapeutic effect and adverse drug reactions, etc. in the era of precision medicine. Furthermore, the development direction of the new model of clinical pharmaceutical care faced on the precision medicine is prospected.