Humans ; Myocardial Reperfusion Injury

Objective To investigate the distribution, isolation and culture of the epidermal stem cells from rats. Methods Immnohistochemical methods were used to confirm the location of the epidermal stem cells. The skin of neonatal rats were dissociated into single cells by dispaseⅡ and trypsin solution,the rapidly adherent cells to collgenⅣ were cultured with KSFM,and those no rapidly adherent cells were regarded as control. Immunohistology and flow cytometry were conducted to identify the epidermal stem cells. Results ? 6-integrin and K15 were expressed in the basal layer cells and hair follicle bulge cells, while the CD71 was negative negatively expressed. CD34 were expressed in the hair follicle bulge cells while not in the basal layer cells. The epidermal stem cells isolated by collgenⅣ had higher colony forming efficiency. Immunocytochemical staining showed that ? 6-integrin and K15 were strongly expressed in the cultured epidermal stem cells. Flow cytometry indicated that 84% cultured epidermal stem cells were expressed ? 6-integrin. Conclusion The epidermal stem cells of rats are located at the basal layer of epidermis and the hair follicle bulge.

ATP-binding cassette protein E (ABCE1) has been annotated as an Rnase L inhibitor in eukaryotes. Previous study showed that the overexpression of ABCE1 was related with the occurrence and clinical stage of lung adenocarcinoma. As an initial investigation into the novel functions of ABCE1, siRNA-expressing vectors targeting sites of the ABCE1 gene were constructed from RNAi-Ready pSIREN-DNR-DsRed-Express vector. Cultured 95-D and NCI-H446 lung carcinoma cells were transfected with the siRNA-expressing vectors using FuGENE 6 and transfection efficiency was determined by using fluorescence microscopy. The expression level of ABCE1 protein was determined by Western blot and immunofluorescence staining. Cell viability was determined by MTT, cell cycle was analysed by flow cytometry.The apoptotic rate was observed by ELISA. Fluorescence microscopy showed a satisfactory transfection efficiency which was about 42.70%. Cell viability and the growth fraction were markedly suppressed, whereas the apoptosis was significantly increased in SiRNA-95-D and SiRNA-NCI-H446 cells than controls(P< 0.05). It can be concluded that the siRNA targeting ABCE1 gene shows a dramatic inhibitory effect on RNA transcription and protein expression and a promoting effect on the apoptosis in 95-D/NCI-H446 cells, which offers a reliable base for the further in vivo experiment.

The Cashmere goat is mainly used to produce cashmere, which is very popular for its delicate fiber, luscious softness and natural excellent warm property. Keratin associated protein (KAP) and bone morphogenetic protein (BMP) of the Cashmere goat play an important role in the proliferation and development of cashmere fiber follicle cells. Bacterial artificial chromosome containing kap6.3, kap8.1 and bmp4 genes were used to increase the production and quality of Cashmere. First, we constructed bacterial artificial chromosomes by homology recombination. Then Tol2 transposon was inserted into bacterial artificial chromosomes that were then transfected into Cashmere goat fibroblasts by Amaxa Nucleofector technology according to the manufacture's instructions. We successfully constructed the BAC-Tol2 vectors containing target genes. Each vector contained egfp report gene with UBC promoter, Neomycin resistant gene for cell screening and two loxp elements for resistance removing after transfected into cells. The bacterial artificial chromosome-Tol2 vectors showed a high efficiency of transfection that can reach 1% to 6% with a highest efficiency of 10%. We also obtained Cashmere goat fibroblasts integrated exogenous genes (kap6.3, kap8.1 and bmp4) preparing for the clone of Cashmere goat in the future. Our research demonstrates that the insertion of Tol2 transposons into bacterial artificial chromosomes improves the transfection efficiency and accuracy of bacterial artificial chromosome error-free recombination.
Animals ; Bone Morphogenetic Proteins ; genetics ; Chromosomes, Artificial, Bacterial ; DNA Transposable Elements ; Fibroblasts ; Goats ; genetics ; Keratins ; genetics ; Transfection

With the development of modern radiotherapy techniques,radiotherapy has been widely used in the multimodality therapy for various malignant tumors,including head and neck cancers such as nasopharyngeal cancer and laryngeal cancer.A combination of surgery and radiochemotherapy significantly improves patients' cure rate and survival time;however,with the increase in survival time,some patients receiving radiotherapy develop marked cognitive impairment.Ionizing radiation-induced cognitive impairment mainly nanifests as hippocampus-dependent cognitive impairment,which is associated with inhibited hippocampal neurogenesis due to ionizing radiation.Therefore,it is necessary to investigate the mechanisms of the inhibition of hippocampal neurogenesis by ionizing radiation.This article reviews the molecular mechanism of neurogenesis disorders induced by ionizing radiation.

Objective: To assess the implementation effects of clinical pathways, compared with usual care, among patients with stroke. Methods:Two investigators independently searched PubMed, Embase, the Cochrane Library, Web of Science, Chinese Biomedical Literature Database and Wanfang Database for studies published before December 2014. Jadad methodological approach was applied to assess the quality of included studies and RevMan software (version 5.2.7) was used for meta-analysis. Results: A total of 11 RCTs involving 913 patients were included in this meta-analysis. The overall results showed that a shorter average length of stay [MD = -2.92; 95% CI (-4.06, -1.78); P < 0.001] and a lower inpatient expenditures [SMD = -1.64; 95% CI (-1.80, -1.48); P < 0.001] in clinical pathways group comparing with the usual care group. The higher score of patient satisfaction was also seen in clinical pathways group. Conclusion: clinical pathways may reduce the average length of stay, reduce the inpatient expenditures, increase patient satisfaction and improve the quality of care in stroke management.

This study examined the cytotoxicity of a new implant material modified by microarc oxidation technique. Cells on different surfaces of the implant were evaluated 2, 4 and 6 days after treatment. The results showed that cell attachment, cell morphology, and cell proliferation were influenced by the different surface treatments, and a significant increase in the osteoblast cell activity was observed on the porous MAO-Ti coating. Our results suggest that the porous MAO-Ti surface has a better biocompatibility and electrochemical performance than pure titanium surface.

Objective: To assess the implementation effects of clinical pathways, compared with usual care, among patients with stroke. Methods:Two investigators independently searched PubMed, Embase, the Cochrane Library, Web of Science, Chinese Biomedical Literature Database and Wanfang Database for studies published before December 2014. Jadad methodological approach was applied to assess the quality of included studies and RevMan software (version 5.2.7) was used for meta-analysis. Results: A total of 11 RCTs involving 913 patients were included in this meta-analysis. The overall results showed that a shorter average length of stay [MD = -2.92; 95% CI (-4.06, -1.78); P < 0.001] and a lower inpatient expenditures [SMD = -1.64; 95% CI (-1.80, -1.48); P < 0.001] in clinical pathways group comparing with the usual care group. The higher score of patient satisfaction was also seen in clinical pathways group. Conclusion: clinical pathways may reduce the average length of stay, reduce the inpatient expenditures, increase patient satisfaction and improve the quality of care in stroke management.
Stroke

Objective To explore the variation of insulin like growth factor-I (IGF-I) and glucose and correlation in children with sepsis. Methods Forty-two children with sepsis in pediatric intense care unit were enrolled from January 2009 to January 2010. In the morning (2nd morning) after admission, the blood glucose, serum IGF-I, cortisol, insulin, IL-6, and IGF-binding protein-I (IGFBP-1) were detected. In the 3rd and 5th morning, the serum IGF-1 was detected again. According to the blood glucose level of the 2nd morning, the children with sepsis were divided into hyperglycemia group and normal group. Meanwhile, 60 healthy children were served as control group. The data had been compared among three groups. Results In the 2nd morning, the levels of blood glucose, serum IGF-I, cortisol, insulin, and IL-6 were signiifcantly different among three groups (all P<0.05), but the serum IGFBP-I was not signiifcantly different (P>0.05). Compared with control group, the sepsis children with hyperglycemia and with normal blood glucose all had signiifcantly higher serum levels of cortisol and IL-6, and signiifcantly lower serum level of IGF-I. In the 2nd, 3rd, and 5th morning, the serum levels of IGF-1 were not signiifcantly changed with time in sepsis children with hyperglycemia and with normal blood glucose (all P>0.05). Meanwhile, there were no signiifcant differ-ences in the serum levels of IGF-1 between sepsis children with hyperglycemia and with normal blood glucose in the 2nd, 3rd, and 5th morning (all P>0.05). In children with sepsis, the blood glucose and serum IGF-1 was not correlated in the next morning (r=0.152, P=0.267). Conclusions The serum level of IGF-I decreased but maintain stable in children with sepsis. The change of blood glucose may be not related with IGF-I.

Objective:To establish a simple HPLC method for the determination of warfarin in human plasma and analyze the re-sults of 100 monitoring reports. Methods:Warfarin was extracted from the plasma samples by acetonitrile. The extract was separated on a ZORBAX Eclipse XDB-C8 column (150 mm × 4. 6 mm, 5 μm)with a mobile phase consisting of methanol and 0. 1% acetic acid (65∶35) at flow rate of 1. 0 ml·min-1 and detected by a diode array detector. The detection wavelength was at 308nm and the sample size was 20 μl. Results:The calibration curve was linear within the range of 0. 1-5. 12μg·ml-1 . The average recovery and RSD was ranged from 85. 0% to 102. 3% and 1. 2% to 3. 9%(n=6), respectively. Inter-and intra-day RSD were 1. 1%-1. 8% and 1. 6%-2. 9%, respectively. Conclusion:The proposed method is rapid, convenient and accurate in the detection of warfarinin concentration in plasma.