Objective To investigate the effects of transfection of chemokine receptor CXCR4 antisense-RNA on the functional expression of VEGF-C mRNA and the invasive ability of colon cell line HT-29 in vitro.Methods PCR primers were designed according to the coding sequence of CXCR4 gene.EcoR Ⅰ and BamH Ⅰ recognition sequences and cutting sites were added to the 5' end of the antisense primer.The purified PCR fragment was retro-inserted into the cloning site of eukaryotic expression vector pcDNA3.1(+).The pcDNA3.1(+)for CXCR4 antisense-RNA was transfected into colon cancer HT-29 cells by liposome transfection.The expression of VEGF-C mRNA was detected by RT-PCR.The expression of CXCR4 protein was determined by Western blot.At the same time,cell growth kinetics was assessed by MTT assay,and in vitro invasive ability was assessed with Boyden chamber.Results CXCR4 antisense-RNA recombinant was successfully constructed.The expression of VEGF-C mRNA in antisense-CXCR4 transfected(HT-29tran)group decreased by 54.2%,while the expression of VEGF-C mRNA in vacant-transfected(HT-29KZ)group only decreased by 9.4% campared with non-transfected(HT-29)group.The difference between the two groups mentioned above was remarkable(P

Objective To investigate the molecular mechanism of metformin. Methods The changes of protein expression of IRS-1 in liver, skeletal muscle and adipose tissues after treatment with metformin for OLETF rats, a model of spontaneous type 2 diabetes mellitus, were measured by Western blot analysis, and compared with those before treatment. Results 22 weeks after the treatment with metformin, the protein expression of IRS-1 was significantly increased in liver (P0. 05) ;and the protein expression of IRS-1 in adipose tissue was significantly decreased (P

Objective To study the influence of unilateral and bilateral amplification on the effect of hearing aid evaluation .Methods Using the subjective method that International Outcome Inventory for Hearing Aids (IOI-HA) and objective method that medium acoustic intensity (65 dB SPL) word recognition score(WRS) to evaluate the effect of unilateral and bilateral hearing aid fitting of middle -aged severe sensorneural hearing loss .Results Hearing aid were used for severe sensorneural hearing loss and the improvement of monosyllables and sentences in quiet and noise test of unilateral were 35 .73% ,43 .15% ,43 .23% ;the improvement of monosyllables and sentences in quiet and noise test of bilateral were 37 .90% ,51 .33% ,54 .86% .The WRS of bilateral was higher than unilater-al .The score of IOI-HA was 15~37 ,meaning patients with severe sensorneural hearing loss were satisfied with hearing aid ,and there was no statistical significance between unilateral and bilateral fitting .Conclusion The bilat-eral hearing aid fitting was better than unilateral .Binaural hearing loss are recommended to fit bilaleral hearing aids .

Animals ; Animals, Newborn ; Blotting, Western ; Brain ; metabolism ; Disease Models, Animal ; Hypoxia, Brain ; metabolism ; Immunohistochemistry ; Myelin Proteins ; genetics ; immunology ; metabolism ; Nogo Proteins ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors

Objective To investigate the effect of purified urinary IgG from patients with minimal change nephrotic syndrome and membranous nephropathy on the expression of macrophage migration inhibitory factor (MIF) in human proximal tubular epithelial cells (HK-2). Methods Proteins were precipitated from urine with (NH4)2SO4, and urinary IgG was purified by protein G column chromatography. SDS-PAGE and Western blotting were performed to analyze the urinary IgG. HK-2 cells were incubated with urinary IgG (0,0.5,1.0,2.5,5.0,10.0 mg/ml) from either minimal change nephrotic or membranous nephropathy patients. The expression of MIF mRNA was assessed by RT-PCR, and MIF protein was assessed by Western blotting. Results SDS-PAGE showed that there were four bands, which were all IgG fractions confirmed by Western blotting. The urinary IgG up-regulated the expression of MIF mRNA and protein in HK-2 cells. The expression increased in a dose-dependent manner. The expression of MIF mRNA and protein in HK-2 cells increased significantly when they were incubated with urinary IgG from membranous nephropathy patients at 1mg/ml (P

Objective To investigate the relationship between the mechanism of ischemic postconditioning-induced activation of nuclear factor-E2 related factor 2 (Nrf2)-antioxidant response element (ARE) signaling pathway during myocardial ischemia-reperfusion (I/R) and reactive oxygen species (ROS).Methods Healthy male Sprague-Dawley rats, aged 16-20 weeks, weighing 250-300 g, were heparinized and anesthetized with intraperitoneal 1％ pentobarbital sodium 40 mg/kg.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution.Thirty-two isolated rat hearts were randomly divided into 4 groups (n=8 each) using a random number table: control group (group C) , group I/R,ischemic postconditioning group (group IPO) , and N-(2-mercaptopropionyl)-glycine (a ROS scavenger) + IPO group (group M + IPO).After 20 min of equilibration, group C was continuously perfused with K-H solution for 100 min, and the isolated hearts received the drugs via the perfusion system in the other groups.Group I/R was perfused with cardioplegic solution 4 ℃ St.Thomas, and then was subjected to 40 min of ischemia at 32 ℃ followed by 60 min of reperfusion.In group IPO, ischemic postconditioning was induced by 6 cycles of 10 s reperfusion followed by 10 s limb ischemia starting from the onset of reperfusion, and the hearts were then perfused for 58 min.In group M + IPO, the hearts were perfused with K-H solution containing N-(2-mercaptopropionyl)-glycine 2 m mol/L for 3 min starting from the onset of reperfusion,underwent 2 min of ischemic postconditioning, and then was perfused for 55 min.Heart rate (HR), left ventricular developed pressure (LVDP), left ventricular end-diastolic pressure (LVEDP),and positive maximal pressure of left ventricular increase (+dp/dtmax) were recorded at the end of equilibration and of reperfusion.At 5 min of reperfusion and the end of reperfusion, myocardial specimens were obtained from the left ventricle for determination of ROS content by enzyme-linked immunosorbent assay.At the end of reperfusion, myocardial specimens were obtained from the left ventricle for examination of the ultrastructure of myocardial cells and for determination of Nrf2, heme oxygenase-1 (HO-1) , quinone oxidoreductase 1 (NQO1), and superoxide dismutase 1 (SOD1) mRNA and protein expression (by using Western blot and real-time polymerase chain reaction).The damage to myocardial mitochondria was assessed using Flameng scoring.Results Compared with group C, HR, +dp/dtmax and LVDP were significantly decreased, and LVEDP was increased at the end of reperfusion in I/R and M+IPO groups, HR and LVDP were decreased, LVEDP was increased, and no significant changes were found in +dp/dtmax at the end of reperfusion in IPO group, Flameng score was increased in I/R, IPO and M+IPO groups , the ROS content was increased at the end of reperfusion in I/R, IPO and M+IPO groups, and Nrf2, HO-1,NQO1 and SOD1 mRNA and protein expression was down-regulated at the end of reperfusion in I/R, IPO and M+IPO groups.Compared with group I/R, HR, +dp/dtmax and LVDP were significantly increased, and LVEDP and ROS content were decreased at the end of reperfusion, Nrf2, HO-1, NQO1 and SOD1 mRNA and protein expression was up-regulated at the end of reperfusion in IPO and M+IPO groups, Flameng score was decreased in IPO group, there was no significant change in Flameng score in M+IPO group.Compared with group IPO, HR, +dp/dtmax and LVDP were significantly decreased, LVEDP and ROS content were increased at the end of reperfusion, Flameng score was increased, and Nrf2, HO-1, NQO1 and SOD1 mRNA and protein expression was down-regulated in M+IPO group.Conclusion Ischemic postconditioning can regulate ROS level and activate Nrf2-ARE signaling pathway, thus attenuating myocardial I/R injury in rats.

Objective To analyze the metabolic profile of children with dyskinetic cerebral palsy by metabolomics, and its abnormal metabolic pathway. Methods The serum of 10 children with dyskinetic cerebral palsy (patient group) and 7 healthy children (control group) aged 6 to 12 years were collected at clinic from May to August, 2014. The serum samples were tested by the nuclear magnetic resonance spectrometer and the spectroscopies were discriminated by partial least squares-discriminant analysis. According to the human metabolome database, the final metabolites disturbed would be figured out. Results 15 chemical shifts were defined, and 6 of them, including 2.04 ppm, 2.12 ppm, 3.00 ppm, 3.24 ppm, 3.76 ppm, 6.50 ppm, were significantly different between 2 groups (P<0.05). The KEGG Pathway Database showed that the levels of taurine, fumarate, oxaloacete, pyruvate, citrate, aspartate, succinate, malate, cysteine decreased, and the levels of glutamate, 2-oxoglutarate, glutamine, leucine, alanine increased. The abnormal metabolism was found in taurine metabolism, glutamine me-tabolism and energy metabolism pathways. Conclusion Based on metabolomics, the metabolic profile of children with dyskinetic cerebral palsy was discriminated out successfully. The further research can focus on the small molecules found out.

Objective To study the effects of different hypoxic training modes on the activities of respiratory chain complexes in the brain mitochondria of rats. Methods Forty healthy two-month-old male Wister rats were randomly divided into 5 groups: living low-training low(LoLo), living high-training high(HiHi), living high-training low (HiLo), living low-training high(LoHi), and living high-exercise high-training low(HiHiLo). Rats were forced to complete a 5-week endurance training program. All training sessions were performed at the same relative intensity under normoxic(P=632mmHg, simulating about 1500m altitude) or hypoxic conditions(P=493mmHg, simulating 3500m altitude) for each of the five groups, respectively. Mitochondria were isolated with differential centrifugation. Spectrophotometric analysis was applied to evaluate RCC (Ⅰ- Ⅲ) activities in brain mitochondria. Results Compared with the LoLo, brain RCC activities in C Ⅱ in rats from LoHi significantly decrease(P<0.05). Brain RCC activities in C Ⅲ in rats from HiHi, HiLo and LoHi significantly decrease and from HiHiLo significantly increase (P<0.01). Conclusion These findings suggest that among different altitude training modes, HiHiLo is the superior one in developing function of mitochondria respiratory chain of rat brain tissues.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; immunology ; pathology ; Cytokines ; blood ; Esophageal Neoplasms ; immunology ; pathology ; Female ; Humans ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-12 ; blood ; Interleukin-4 ; blood ; Interleukin-5 ; blood ; Lymphatic Metastasis ; Male ; Middle Aged ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Tumor Necrosis Factor-alpha ; blood

By means of literature review,expert interviews and experts sorting,the authors identified three approaches of medical humanities care,and applied them to three clinical departments of top ten incidence of medical disputes of the hospital in 2012.These departments have similar number of beds,and covered both internal medicine and surgery departments.The purposes of the study are to evaluate the effects of medical humanities care measures in clinical departments,and to make relevant thoughts and suggestions.