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.

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

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.

BACKGROUND:Liushen Pil is a traditional Chinese herbal formula, has the effects of heat-clearing and detoxicating, eliminating stagnation, detumescence and al eviating pain. Modern pharmacology verifies that Liushen Pil has anti-inflammatory, analgesic, cardiac, anti-viral, anti-tumor effects, and has been extensively used in the treatment of various infectious diseases and malignant cancer. OBJECTIVE:To investigate the inhibitory effects of Liushen Pil on esophageal cancer xenografts, and effects on microvessel density and vascular endothelial growth factor expression. METHODS:After reproducing nude mouse models of human esophageal cancer, 48 nude mice were randomly divided into high-dose Liushen Pil group, moderate-dose Liushen Pil group, low-dose Liushen Pil group, cisplatin group, model group and blank group. According to medication regimen, drugs were given. The growth of transplanted tumor of nude mice was dynamical y observed in each group. The nude mice were sacrificed after 20 days of treatment. Neoplasm weight was taken and the tumor-suppressing rate was calculated. Immunohistochemistry was used to detect microvessel density and the expression of vascular endothelial growth factor. RESULTS AND CONCLUSION:The weight of transplanted tumor was significantly lower in the high-dose Liushen Pil group, moderate-dose Liushen Pil group, low-dose Liushen Pil group, and cisplatin group than in the model group (P<0.05). Microvessel density and the expression of vascular endothelial growth factor were obviously lower in the each Liushen Pil group than in the model group, but not as apparent as that in the cisplatin group. Results suggested that Liushen Pil can inhibit the growth of the esophageal cancer xenografts. Liushen Pil can down-regulate the expression of vascular endothelial growth factor and reduce microvessel density, which is one of the tumor-inhibiting mechanism of Liushen Pil .

In this article, the method of quick and accurate identification ofAstragalus membranaceus (Fisch.) Bge. var.mongholicus (Bge.) Hsiao in different growth years by electronic nose (e-nose) technique and chemical analysis was developed. An e-nose was used to detect the odors ofA. membranaceus. var.mongholicus samples in two-growth-year, seven-growth year and more than a decade growth years for establishing the classification model of response characteristics. Principal component analysis (PCA) and discriminant factor analysis (DFA) were performed to differentiate theA. membranaceus. var.mongholicus samples in different growth years. PCA was performed to investigate the difference in the chemical composition and quality of different growth years inA. membranaceus. var.mongholicus samples. The results of PCA and DFA analyses for e-nose demonstrated that the samples ofA. membranaceus. var.mongholicus in different growth years could be distinguished obviously. The contents of chemical composition were similar in same growth years inA. membranaceus. var.mongholicus and different from different growth years. The results of chemical composition analysis indicated that the identification forA. membranaceus. var.mongholicus in different growth years was significant for the quality control. E-nose technique could identify the samples ofA. membranaceus. var.mongholicus in different growth years rapidly, sensitively and intactly, and could be applied for the quality control of traditional Chinese medicine.

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.

The chronic myeloid leukemia( CML) is characterized by a cytogenetic abnormality.The BCR-ABL fusion gene encodes protein 210.With the rapid development of molecular biology and other technologies, the treatment of CML has made great progress.However,patients for TKI resistance,which cannot be tolerated,and TKI will not eliminate CML stem cells.Despite hematopoietic stem cell transplantation( HSCT) is recommended as first-line treatment,it is still faced with many problems.Therefore,to clear CML tiny residual lesions from Ph+malignantly clonal stem cells has become an urgent need,which is expected to be an effective method for CML patients to obtain permanent cure and long-term disease-free survival.In this paper,we review the main advances achieved in the treatment of CML.

Objective To investigate the variation of interleukin-6 ( IL-6 ) , vascular endothelial growth factor(VEGF),E-selectin and intercellular adhesion molecule-1(ICAM-1)in children with sepsis and the clinical significance. Methods This was a prospective and control study. Thirty-two children diagnosed as sepsis in PICU from December 2008 to December 2009 served as the sepsis group. According to whether there was a shock, sepsis group were divided into shock subgroup and no shock subgroup. Fifteen healthy children served as control group. The serum levels of IL-6,VEGF,E-selectin and ICAM-1 were detected with enzyme linked immunosorbent assay. Results The serum level of IL-6 was 65. 00(30. 49~237. 14) ng/L in shock subgroup and 48. 68(30. 25~75. 00) ng/L in no shock subgroup,which were significantly higher than that in control group[0. 80(0. 60 ~1. 00) ng/L](P<0. 05). There was no significant difference between shock subgroup and no shock subgroup. The serum levels of VEGF and E-selectin showed no significant differences among the three groups. The serum level of ICAM-1 was 998. 72(666. 93~1 526. 44) ng/ml in shock subgroup,and 925. 71(683. 53~1 225. 12) ng/ml in no shok subgroup,which were significantly high-er than that in control group[660. 59(525. 48~685. 47) ng/ml]. Compared with those who survived in sep-sis group,the serum levels of VEGF and E-selectin in the died children with sepsis showed no significant difference,but IL-6 and ICAM-1 significantly increased(P<0. 05). Conclusion IL-6 and ICAM-1 increase greatly and accentuate inflammation in septic patients,the changes of which may help to determine the prog-nosis of sepsis.

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.