Objective To examine the effects of oxidative stress induced damage to the Prestin expression in HEI-OC1 cells,and to study the mechanism of sensory deafness.Methods We used different concentrations (50μM,100μM,200μM)of hydrogen peroxide canister to cultivate HEI-OC1 cells,and to detect the activity of su-peroxide dismutase(SOD).The quantitative real-time PCR and immunofluorescence were used to detect the prestin expression of mRNA.Results The SOD activity decreased in the HEI-OC1 cells damaged by oxidative stress.The high concentration of the infected group decreased more significantly(F= 9926.293,P<0.01).The expressions of Prestin mRNA and Prestin protein were decreased obviously in the HEI-OC1 cells.The high concentration of in-fected group decreased more significantly (F= 4065.046and7657.217,P<0.01).Conclusion Oxidative stress in-ducing damage inhibits the expression of prestin.Prestin protein may be used as a molecular marker of sensory deafness.

Adolescent ; Child ; Child, Preschool ; Cytokines ; blood ; Female ; Humans ; Infant ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Male ; Seizures, Febrile ; immunology ; Tumor Necrosis Factor-alpha ; blood

Humans ; Pediatrics ; manpower ; Personnel Administration, Hospital ; Respiratory Therapy Department, Hospital ; manpower

Female ; Humans ; Infant ; Infant, Newborn ; Male ; Stomach Neoplasms ; Teratoma

Background and purpose:The incidence of hepatoma is high. The outcome of treatment on hepatoma is poor.So we investigated the effect and mechanism of a selective cyclooxygenase-2 inhibitor celecoxib on the proliferation and apoptosis of SMMC-7721 hepatoma cell line. Methods:MTT assay was used to study the inhibitive effect of celecoxib on the growth of SMMC-7721 hepatoma cell. The effect of celecoxib on cell cycle and apoptosis on cells was studied by flow cytometry(FCM).Transmission electron microscopy (TEM) was used to display the morphological change of the SMMC-7721 hepatoma cell . The biochemical character of apoptosis was viewed on the agarose gel electrophoresis.The expression of bax gene and bcl-2 gene were measured by immunohistochemistry.Results:The SMMC-7721 cells were cultured in media that contained 25,50,75,100 ?mol/L celecoxib,by means of MTT, the inhibition rate was(15?3)%,(34.6?2.4)%,56.8?1.0)%,(86.2?0.4)% respectively after 24 hours; but the inhibition rate was (33.4?0.7)%,(66.7?1.8)%,(76.1?2.4)%,(97.3?0.8)% respectively after 48 hours(P

Objective To clone human vacuolar protein sorting 4A gene(hVPS4A)and to construct its eukaryotic expressive plasmid.Methods Primers were designed to amplify the full length hVPS4A by PCR using cDNA of Huh7 cell as a template,then the target DNA was inserted into the eukaryotic vector pRK5.The recombinant plasmid was confirmed by PCR,restriction enzyme digestion and DNA sequencing.Results A 1 300 bp fragment was successfully amplified by PCR from the cDNA of Huh7 cells.Af-ter recycled,purified and ligated with the vector pRK5,the recombinant plasmid was transformed into E.coli DH5α.The positive re-combinant plasmid identified by PCR was selectred and digested by EcoRⅠto get a 5 900 bp fragment;and two fragments including 4 600 bp and 1 350 bp were obtained using EcoRⅠand HindⅢ digestion;the size of these two fragments were consistent with the pRK5 target fragment and the inserted hVPS4A as expected.Moreover,DNA sequencing results confirmed that the inserted frag-ment was in accordance with the hVPS4A reference sequence.Conclusion The eukaryotic expression vector containing hVPS4A gene is constructed successfully,which provides the condition for further study on the hVPS4A biological functions.

Background Recently,there were many studies on corneal innervations during mammalian development.However,there were fewer studies on discussing corneal innervations before and after mouse eye openings.Objective The present study was to investigate the change in the regulation of corneal nerve fiber length and density before and after mouse eye openings to offer a basis for clinical research in human.Methods Thirty SPF C57BL/6 mice were divided into postnatal 1 day(P1 d),P7 d,P13 d(1 day before eye opening),P14 d(eye halfopened),P17 d(1 day after eye opening)and P23 d(7 day after eye opening)groups,with 5 mice and 10 eyes for each group.Entire corneal stretches were prepared and immunostaining with an anti-neuron-specific β-Ⅲ tubulin antibody was performed to label the corneal nerve fibers.Confocal microscopic pictures from the corneal dorsal-nasal region (DN),dorsal-temporal(DT),ventral-nasal region(VN)and ventral-temporal(VT)were taken using Delta Vision Core.From these pictures,the mouse corneal area,total length and density of nerve fibers in the 4 regions were calculated.The use of the animals complied with Statement of ARVO.Results Corneal areas of P1 d,P7d,P13 d,P14 d,P17 d and P23 d mice were(0.404±0.007),(1.362±0.154),(1.573±0.080),(1.603±0.046),(1.847±0.052),(2.445±0.798)mm2,respectively ; the total lengths of nerve fibers were(3.718±1.044),(19.065±3.350),(23.687±0.907),(27.309±2.477),(31.989±3.976),(41.214±1.573)mm,respectively ; the densities of nerve fibers were(9.592±1.138),(14.506±1.908),(15.088±1.241),(16.772±1.897),(16.821±2.102),(17.660±1.216)mm/mm2,respectively,all showing significant increases with age(F =22.906,P =0.000 ; F =0.424,P =0.000 ; F =2.375,P=0.000).A positive correlation of the increasing corneal areas and increasing lengths of nerve fibers was found(r=0.983,P＜0.01).Nerve fiber densities in the four corneal regions significantly increased with age(DN region:F =0.159,P =0.000 ; DT region:F =2.1 72,P =0.001 ; VN region:F =1.998,P =0.000 ; VT region:F=2.352,P=0.000).From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0％ without significant difference(t =0.589,P =0.572); and the corneal nerve fiber densities in the DT region,VN region and VT region decreased by 4.6％,5.5％ and 0.1％,respectively,without significant difference from P14 d to P17 d(t=0.549,P=0.596;t=0.701,P=0.501 ;t=-0.100,P=0.919).Conclusions The development of nerve fibers in the whole cornea or the four corneal regions is influenced by eye opening in mouse to various extents.From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0％ without significant difference.From P14 d to P17 d,the corneal nerve fiber densities in the DT region,VN region and VT region decrease by 4.6％,5.5％ and 0.1％,respectively,without significant difference.Afterwards,the growth of nerve fibers increased in pace and the growth rate is recovered.

OBJECTIVE: To analyze the causes of the esophagogastric junction outlet obstruction (EGJOO) patients, to discuss the differences of the clinical manifestation and esophageal motility characteristics between the anatomic EGJOO (A-EGJOO) and functional EGJOO (F-EGJOO) subgroups, and to search the diagnostic values of the specific metrics for differentiating the subgroups of EGJOO patients. METHODS: For the current retrospective study, all the patients who underwent the esophageal high resonance manometry test were retrospectively analyzed from Jan 2012 to Oct 2018 in Peking University Third Hospital. The EGJOO patients were enrolled in the following research. The clinical characteristics, such as symptoms and causes of the patients were studied. Then the patients were divided into two subgroups as A-EGJOO subgroup and F-EGJOO subgroup. The clinical symptoms and the main manometry metrics were compared between these two subgroups. The significant different metrics between the two groups were selected to draw receiver operating characteristic (ROC) curves and the diagnostic values were analyzed in differentiating the A-EGJOO and F-EGJOO subgroups. RESULTS: The most common symptom of EGJOO was chest pain or chest discomfort (30.63%), then the dysphagia (29.73%), and acid regurgitation/heartburn (27.03%). Non-erosive reflux disease (36.04%) was the most popular cause for EGJOO, then the reflux esophagitis (17.12%). Besides the intra-EGJOO and extra-EGJOO lesions, the connective tissue disease (6.31%) and central nervous diseases (2.70%) were found to be the etiology of EGJOO. The causes of the rest 19 EGJOO were unknown. A-EGJOO patients presented significantly higher intra bolus pressure (IBP) than that of F-EGJOO [6.80 (5.20, 9.20) mmHg vs. 5.10 (3.10, 7.60) mmHg, P=0.016]. The area under curve of IBP was 0.637. When IBP≥5.15 mmHg, the sensitivity was 78.60% and specificity 50.70% to differentiate A- or F-EGJOO. CONCLUSION Chest pain or chest discomfort was the most common symptom in EGJOO patients. Besides the intraluminal structural disorders, the extra-luminal causes were found in EGJOO patients. A-EGJOO presented higher IBP than that of F-EGJOO patients. The cutoff value of IBP to differentiate A-EGJOO from EGJOO was 5.15 mmHg with sensitivity 78.06% and specificity 50.70%. However for the low area under curve, the diagnostic value of IBP was limited.
Deglutition Disorders ; Esophageal Motility Disorders/diagnosis* ; Esophagogastric Junction ; Humans ; Manometry ; Retrospective Studies

Human herpes simplex virus 1 (HSV-1) causes facial,ocular,and encephalitic disease and is associated with latent infection and cancer.Here,we developed a means of studying the pathogenesis of HSV-1 infection at the protein level by using the SELDI Protein Chip to detect changes of protein expression in Vero cells cultured in vitro.After infection with HSV-1 and culture for 12,24 or 48 h,cells were harvested and lysed.IMAC3 arrays were applied to SELDI-TOF-MS to detect proteomic differences before and after infection.The chip detected a series of differentially expressed protein peaks.Interestingly,both peaks at 16 912 Da and 17 581 Da corresponded precisely with the molecular mass of ISG 15,which may participate in antiviral activity during the process of infection.Thus,the results we obtained can serve as a basis to study the pathogenesis of HSV-1 and the interaction between the virus and its host.In addition,they can help in the discovery of new therapeutic targets for treatment of HSV-1 infection.

Objective To study the effect of aerobic exercise and dietary intervention on lipid metabolism in metabolic syndrome rats, and investigate the possible mechanism mediated by peroxisome proliferator-activated receptorα(PPARα). Methods After one-week feed-ing, Sprague-Dawley rats were randomly divided into blank control group (CC group) and model group which were feed in high-fat-and-salt diet for 18 weeks to establish a metabolic syndrome model. Then, the metabolic syndrome rats were randomly divided into model control group (MC), the model high-fat diet group (MHE) and the model general died exercise group (ME). ME and MHE groups were forced to run on a treadmill for twelve weeks at the same time. The weight of perirenal fat, blood free fat acid (FFA), and blood lipid were determined. The expression of PPARαmRNA in myocardium was detected by RT-PCR. Western blotting was applied to detect the protein expression of PPARαin myocardium. Results Compared with CC group, MC group showed significant increase in body weight, perirenal fat weigh, FFA, and blood lipid (P<0.05), and significant decrease in PPARαmRNA and protein expression (P<0.01) in myocardium. Compared with MC group, ME and MHE groups showed significant decrease in body weight, perirenal fat weight, triglyceride (TG), and showed significant in-crease in high-density lipoprotein (HDL), and the expression of PPARαmRNA and protein in myocardium (P<0.05). Compared with MHE group, ME group showed decrease in low-density lipoprotein (LDL) (P<0.05), and increase in the expression of PPARαmRNA and protein (P<0.01). Conclusion Aerobic exercise may activate the expression of PPARα, enhance the utilization of fatty acid, reduce body mass and visceral fat mass, improve the dyslipidemia and then regulate lipid metabolism in metabolic syndrome rats.