Objective To explore the effects of music intervention on growth and development of premature infants. Methods 112 premature infants were randomly divided into the observation group and the control group with 56 cases in each. The control group received conventional nursing measures, and the observation group was simultaneously given music intervention besides the conventional routine nursing measures. Weight increase, hospitalization time, milk-intake volume and feeding tolerance were compared between the two groups. Results The weight increase in the observation group was higher than that in the control group. Hospitalization time and rate of feeding intolerance for the observation group was lower compared with that of the control group. The premature infants in the observation group took in more milk than those in the control group. Conclusions Music intervention can elevate feeding tolerance, facilitate nutrition and increase weight of premature infant, so it is beneficial to the growth and development of premature infant.

AIM To discuss the effects of losartan on AT 1R mRNA expression and renin angio tensin system (RAS) level in brain of spontaneously hypertensive rat (SHR). METHODS Sixteen 6 week old male SHR and 16 sex and age matched WKY were divided into losartan ( n =8) and control groups ( n =8) respectively. Losartan (30 mg?kg -1 ?d -1 ) or equal volume of 0.9% saline solution was administered for 18 weeks by gavage. AT 1R mRNA expression was examined by reverse transcription polymerase chain reaction (RT PCR). Renin activity (RA) and angiotensinⅡ(AngⅡ) levels were measured by radio immunoassay. Angiotensin converting enzyme activity (ACEA) was measured by ultraviolet spectrophotometar. RESULTS Enhanced brain AT 1R mRNA expression was found in SHR compared with WKY ( P 0.05, respectively). CONCLUSION The increase of AT 1R mRNA expression and RAS levels in brain may be involved in the pathogenesis of hypertension in SHR. Losartan protects brain by decreasing expression of brain AT 1R mRNA.

Objective To study the Ile796Val polymorphism in human SCAP gene in Hubei area, and analyze its correlation with coronary heart disease (CHP) and hypertension and the relationship between polymorphism and lipid metabolism. Methods Using PCR RFLP, we detected genotypes of Ile796Val polymorphism in human SCAP gene. Results The allele A frequencies of Ile796Val in human SCAP gene for controls, CHD patients and the hypertension patients were 0.32, 0.45 and 0.42 respectively. The allele G frequencies were 0 68, 0.55 and 0.58 respectively. There were significant differences in frequencies of genotype and alleles between controls and hypertension group. And there was significant difference in the level of TC, LDL C and ApoB. In CHD group, there was significant difference in the TC level between different genotypes. In hypertension patients, although a difference was noted in genotype, there was no significant difference in allele frequencies and lipid level exceps a significant difference in the levels of TC, LDL C and ApoB in hypertension patients. Conclusion Ile796Val polymorphism in human SCAP gene may be a great agent to cause disorder in the lipid level of blood and lipid metabolism of tissue. It is of great significance in disorder in lipid metabolism of inter cellular and genetic investigation of hyperlipidemia.

Background Krüppel-like factor 6 (KLF6) is related to the physiological or pathological process,such as growth,cell differentiation,proliferation,apoptosis,angiogenesis,tissue repair,and so on.But in ophthalmology,it is less reported about the expressing level of KLF6 protein in lens epithelial cclls (LECs) or the effect of KLF6 on the proliferation of human LECs.Objective This study was to investigate whether KLF6 can inhibit proliferation of human LECs.Methods KLF6 eukaryotic expression plasmid (pEGFP-C2-KLF6) was constructed using reverse transcription PCR(RT-PCR) and identified by double enzyme digestion method and PCR.Human LECs strain (HLE-B3) was cultured and passaged using low glucose DMEM containing 10％ fetal bovine serum and then divided into 4 groups.KLE-B3 transfection reagents were added in the culture medium of all groups.In addition,no agent was used in the blank control group;only insulin-like growth factor-1 (IGF-1) was appended to the medium in only IGF-1 group ;null vector was transfected and IGF-1 was appended in the null plasmid transfection+ IGF-1 group;while pEGFP-C2-KLF6 eukaryotic expression plasmid was transfected into the cells,and simultaneously IGF-1 was added in the pEGFP-C2-KLF6 plasmid transfection+IGF-1 group.After 24 hours of intervene,water soluble tetrazolium salt-1 (WST-1) test was used to detect the growth status of the cells,and Western blot assay was used to assay the relative expressing level of KLF6 protein in the cells.In the other hand,the cells were cultured at the density of 1 ×104/piece,and 0,0.10 and 0.25 μg pEGFP-KLF6 were transfected into each piece of cells respectively,and then IGF-1 was added with a final concentration of 50 μg/L for 24 hours after cell culture.Expressions of Ki-67 protein and mRNA in the cell pieces were detected by immunocytochemistry and fluorescent semiquantitative PCR,respectively.Results The PCR product bands were consistent with KLF-6 gene in length,and the product fragments were corresponding with expectant ones via PCR and double enzyme digestion method,showing a successful construction of pEGFP-C2-KLF6 eukaryotic expression plasmid.After 24 hours of IGF-1 stimulation,the absorbance values of the cells were 0.86±0.00,2.10±0.01,2.24±0.12 and 1.06±0.02 in the blank control group,only IGF-1 group,null plasmid transfection + IGF-1 group and the pEGFP-C2-KLF6 plasmid transfection + IGF-1 group,with a significant difference among the 4 groups (F =38.322,P ＜ 0.05),and that in the pEGFP-C2-KLF6 plasmid transfection +IGF-1 group was significantly lowed in comparison with the only IGF-1 group and the null plasmid transfection+IGF-1 group (q=6.42,7.31,both at P＜0.05).Western blot assay showed that the relative expressing levels of KLF6 protein were statistically different among the four groups (F =591.858,P＜0.05),and those in the pEGFP-C2-KLF6 plasmid transfection+IGF-1 group were 1.47,2.04,3.27 folds higher than those in the blank control group,only IGF-1 group,respectively and null plssmid transfection+IGF-1 group,respectively.Immunocytochemistry revealed that the expressing intensity of Ki-67 protein was gradually weakened with the decrease of pEGFP-C2-KLF6 plasmid dose in the 0,0.10 and 0.25 μg pEGFP-C2-KLF6 plasmid transfection+IGF-1 groups.Fluorescence semiquantitative PCR results showed that the relative expression values of Ki-67 mRNA in the cells were O.15±0.08 and 0.11±0.03 in the 0.10 μg and 0.25 μg pEGFP-C2-KLF6 plasmid transfection+ IGF-1 groups,which were significantly lower than O.77± 0.12 of the 0 μg pEGFP-C2-KLF6 plasmid transfection group,with a statistically significant difference among the three groups (F =54.825,P＜0.05).Conclusions KLF-6 can effectively inhibit IGF-1-induced proliferation of human LECs,and it can be regarded as one of the regulatory factors of the proliferation of HLE-B3 cells.

Objective To observe the effects of dexamethasone (DEX) therapy on the clinical outcome and changes of lymphocyte subsets in children with mild and severe encephalitis.Methods Eighty-two children with mild and severe viral encephalitis were randomly divided into two groups: with DEX treatment (n=46) or without(n=36).The clinical efficacy was evaluated 3 weeks later, and the clinical manifestation were also observed. The changes of CD4~+,CD8~+ T lymphocyte percentage were determined by flow cytometry on admission and at 1 week,4 weeks after treatment.Another group of 20 cases was enrolled as control group.Results Compared with the control group,both the DEX treatment group and non-DEX treatment group showed a reduced CD4~+ lymphocyte count, an increased CD8~+ lymphocyte count(P

Humans ; Macrophage Activation Syndrome ; genetics ; therapy

Red-based recombineering has been widely used in Escherichia coli genome modification through electroporating PCR fragments into electrocompetent cells to replace target sequences. Some mutations in the PCR fragments may be brought into the homologous regions near the target. To solve this problem in markeless gene deletion we developed a novel method characterized with two-step recombination and a donor plasmid. First, generated by PCR a linear DNA cassette which comprises a I-Sec I site-containing marker gene and homologous arms was electroporated into cells for marker-substitution deletion of the target sequence. Second, after a donor plasmid carrying the I-Sec I site-containing fusion homologous arm was chemically transformed into the marker-containing cells, the fusion arms and the marker was simultaneously cleaved by I-Sec I endonuclease and the marker-free deletion was stimulated by double-strand break-mediated intermolecular recombination. Eleven nonessential regions in E. coli DH1 genome were sequentially deleted by our method, resulting in a 10.59% reduced genome size. These precise deletions were also verified by PCR sequencing and genome resequencing. Though no change in the growth rate on the minimal medium, we found the genome-reduced strains have some alteration in the acid resistance and for the synthesis of lycopene.
Chromosomes, Bacterial ; genetics ; DNA ; Endonucleases ; metabolism ; Escherichia coli ; genetics ; Genetic Engineering ; methods ; Recombination, Genetic ; Sequence Deletion

Brain Injuries ; etiology ; Child ; Heart Defects, Congenital ; complications ; Humans

Objective To evaluate in vivo tracking of swine mesenchymal stem cells (MSCs) la-beled with super paramagnetic iron oxide (SPIO) in intraportal transplantation by a clinical 1.5T MR.Methods MSCs were isolated from swine and cultured as well as expanded, which were then incuba-ted with SPIO (Feridex I. V.). Prussian blue staining was performed for showing intracelluar irons.To establish a swine model of acute liver necrosis, 0.5 g/kg of D-galactosamine was administrated to 10 pigs. MSCs(labeled cells in six, unlabeled cells in four)were injected into liver via portal veins. MR imaging was performed with a clinical 1.5T MR immediately before and at 6 h, 3 d, 7 d, 14 d after transplantation, respectively. Results Prussian blue staining of SPIO labeled MSCs could be effec-tively labeled and the labeling efficiency was almost 100%. Signal intensity loss in liver by SPIO labe-ling on FFE sequence persisted until 14 days after transplantation. Histological analysis by Prussian blue staining showed homing of labeled MSCs in liver after 14 days, primarily distributing in hepatic sinusoids and liver parenchyma. Conclusion MSCs can be labeled with SPIO in vitro successfully.MRI can monitor magnetically labeled MSCs transplanted into liver.

Although significant advances have been made in both the development of therapeutic strate-gies and the understanding of pathophysiological mechanisms of sepsis, the mortality of severe sepsis and septic shock still remains unacceptably high worldwide. Current prediction models based on socio-demo-graphic and clinical risk factors fail to explain fully why a particular patient either develops or succumbs to sepsis. In recent years epidemiological studies have suggested a strong genetic relationship on the suscepti-bility and outcome of sepsis. With the completion of Human Genome Project and International HapMap Pro-ject, the identification of susceptibal genes contributing to sepsis may allow more precise use of interven-tions, such as targeted therapy of sepsis is an appealing strategy. In this review, we summarize a broad over-view of genetic nomenclature, study designs, and problems of these genetic association studies.