The aim of the present study was to investigate the role of glutamate receptors in the damage of spiral ganglion neurons (SGNs) induced by acute acoustic noise. This investigation included in vivo and in vitro studies. In vivo, kynurenic acid (KYNA), a broad-spectrum antagonist of glutamate receptors, was applied to the round window of guinea pigs, and its protective effect was observed. The animals were divided into three groups: control (saline, 0.9%, 10 microL), saline (0.9%, 10 microL) + noise and KYNA (5 mmol/L, 10 microL) + noise. Saline and KYNA were applied to the round window membrane with a microsyringe. The animals were exposed to 110 dB SPL of white noise for 1 h. Hearing thresholds for auditory brainstem responses (ABRs) and compound action potentials (CAPs) in all animals were measured before and after treatment. The amplitudes of III waveform of ABR and N1 waveform of CAP and the latency of N1 waveform at different stimulation levels (intensity-amplitude and intensity-latency functions) were also measured. The cochleas were then dissected for transmission electron microscopy (TEM) after final electrophysiological measurement. In vitro, the SGNs of the normal guinea pigs were isolated and glutamate (100 micromol/L or 1 000 micromol/L) was added into the medium. The morphology of the SGNs was examined by light microscopy. In vivo results showed that the hearing function and morphology of the inner ear including hair cells and SGNs in the control group were normal. Compared with that in the control group the thresholds for ABR and CAP (click and tone burst) in saline + noise group were elevated significantly. The input-output functions showed that the amplitudes of III waveform of ABR and N1 waveform of CAP decreased and the latency of N1 waveform increased obviously. There was significant difference in the amplitude and latency between saline + noise group and KYNA + noise group (P<0.05). TEM indicated obvious swelling and vacuoles at the terminate of dendrites of SGNs in NS + noise group. On the contrary, the afferent dendrites in KYNA + noise group showed normal appearance without swelling and vacuoles. In vitro experiment showed that the isolated SGNs of guinea pigs obviously swelled and even died after application of 100 micromol/L or 1 000 micromol/L glutamate. These results suggest that noise exposure causes hearing impairment, damage of hair cells and hair cell/afferent synapse and death of SGNs. The antagonist of glutamate receptors provides protective effects against hearing loss and SGN damage. It is inferred that excessive release of glutamate from the inner hair cells induced by noise may be responsible for these damages. Glutamate receptors are involved in the degeneration and death of SGNs.
Action Potentials ; physiology ; Animals ; Evoked Potentials, Auditory, Brain Stem ; physiology ; Excitatory Amino Acid Antagonists ; pharmacology ; Guinea Pigs ; Hearing Loss, Noise-Induced ; metabolism ; pathology ; physiopathology ; Kynurenic Acid ; pharmacology ; Male ; Neurons ; pathology ; Noise ; adverse effects ; Random Allocation ; Receptors, Glutamate ; metabolism ; Spiral Ganglion ; pathology

Bladder cancer is one of the most common types of cancer. Most gene mutations related to bladder cancer are dominantly acquired gene mutations and are not inherited. Previous comparative transcriptome analysis of urinary bladder cancer and control samples has revealed a set of genes that may play a role in tumor progression. Here we set out to investigate further the expression of two candidate genes, centromere protein U (CENPU) and mitochondrial ribosomal protein s28 (MRPS28) to better understand their role in bladder cancer pathogenesis. Our results confirmed that CENPU is up-regulated in human bladder cancer tissues at mRNA and protein levels. Gain-of-function and loss-of-function studies in T24 human urinary bladder cancer cell line revealed a hierarchical relationship between CENPU and MRPS28 in the regulation of cell viability, migration and invasion activity. CENPU expression was also up-regulated in in vivo nude mice xenograft model of bladder cancer and mice overexpressing CENPU had significantly higher tumor volume. In summary, our findings identify CENPU and MRPS28 in the molecular pathogenesis of bladder cancer and suggest that CENPU enhances the progression of bladder cancer by promoting MRPS28 expression.

Bladder cancer is one of the most common types of cancer. Most gene mutations related to bladder cancer are dominantly acquired gene mutations and are not inherited. Previous comparative transcriptome analysis of urinary bladder cancer and control samples has revealed a set of genes that may play a role in tumor progression. Here we set out to investigate further the expression of two candidate genes, centromere protein U (CENPU) and mitochondrial ribosomal protein s28 (MRPS28) to better understand their role in bladder cancer pathogenesis. Our results confirmed that CENPU is up-regulated in human bladder cancer tissues at mRNA and protein levels. Gain-of-function and loss-of-function studies in T24 human urinary bladder cancer cell line revealed a hierarchical relationship between CENPU and MRPS28 in the regulation of cell viability, migration and invasion activity. CENPU expression was also up-regulated in in vivo nude mice xenograft model of bladder cancer and mice overexpressing CENPU had significantly higher tumor volume. In summary, our findings identify CENPU and MRPS28 in the molecular pathogenesis of bladder cancer and suggest that CENPU enhances the progression of bladder cancer by promoting MRPS28 expression.

Objective To analysis the E protein epitopes of dengue virus type 1-4, Japanese encephalitis virus and yellow fever virus and to distinguish the shared or specific epitopes among them. Methods Bioinformatic software DNAStar was used to analyze the hydrophilicity, flexibility, surface probability and antigenicity of dengue virus type 1-4, Japanese encephalitis virus and yellow fever virus E prtein amino acid sequences. The influence of secondary structure was also considered. Based on the bio-informatic analysis of E protein epitopes, 6 specific epitopes were amplified and inserted into prokaryotic expression vector pMAL-c2x. The vectors was then transferred into E.coli BL21 (DE3) and Rosetta (DE3). Isopropyl-β-D-thiogalactoside (IPTG) was used to induce the expression of gene segments and SDS-PAGE were used identify the expression proteins. The antigenieity was tested, using Western blot. Results 15 shared epitopes and 47 specific epitopes were forecasted by bioinformatic analysis, and 6 specific epitopes from dengue virus type 1-4, Japanese encephalitis virus and yellow fever virus E protein were expressed in E.coli successfully. Two specific antigenic determinant from dengue virus type 1 and dengue virus type 2 were confirmed using Western blot, while the others epitopes shown no antigenic reaction property. Conclusion Two specific antigenic determinant were confirmed, under Western blot.

OBJECTIVETo establish an efficient and stable method for protein extraction of Streptococcus mutans.

METHODSThe collected bacteria were treated by freeze-thaw and ultrasonic (method 1), ultrasonic (method 2), boiling (method 3), boiling and ultrasonic (method 4), respectively. The index such as state of bacteria broken, concentration of extracted protein and SDS-PAGE of protein were employed to evaluate the effects of above four methods.

RESULTSBeside the method 3, the other three methods could break the bacteria effectively, of which ultrasonic was the key factor. The pattern of SDS-PAGE which treated by method 1, method 2 and method 4 was almost same, but method 1 resulted in the best abundance. There was significantly difference among the four protein concentration extracted by four methods (P < 0.05). All methods exhibited good stability and reproducibility.

CONCLUSIONMethod of freeze-thaw and ultrasonic resulted in an efficient proteins extraction of Streptococcus mutans which demonstrated good stability and reproducibility and easy to handle.

Objective To investigate the differences in genotypes and phenotypic parameters of β-thalassemia gene carriers in pregnant women′s from Chengdu, Sichuan Province.Methods Totally 320pregnant women′s withβ-thalassemia gene from March 2016to June 2017in our hospital were selected.Routine blood tests, alkaline hemoglobin electrophoresis and routine analysis ofβ-thalassemia were performed on all the cases.Statistical analysis was performed on the data of each group.Results There were 306cases of heterozygousβmutations and 10types of mutations, among which 14cases ofα-thalassemia combined had 6types of mutations.The mutations of MCV, MCH, MCHC, and Hb in the routine blood tests of each group showed some differences.The incidence of abnormal bands was also different for each mutation, and the hemoglobin electrophoresis results ofβEM mutations contained abnormal bands.However, the clinical manifestations of CAPM mutations were not obvious and easily missed.Conclusion There is a certain regional specificity inβthalassemia gene carrying in Chengdu area.Targeted examination in the preliminary screening and prenatal diagnosis should be conducted so as to reducing the birth rate of children′s with severe thalassemia.

To increase the productivity and yield of recombinant protein in continual perfusion processing, Every amino acid consumption rate in continual perfusion culture of engineering CHO cell line which expressed recombinant TNFRp75: Fc fusion protein were analyzed. Then rational amino acids were accordingly added to improve its comprehensive utilizing. At the same time, glucose supply was controlled to make the concentration of glucose below 0.5 g/L for ameliorating the toxicity of lactate accumulation in order to decrease the perfusion rate. The result showed that the productivity of recombinant protein was 3.1 times (388mg/L) and the total yield was 4. 7 times (244. 4g) that of control cultures after nutrient compensation and metabolism control in 30 liter working volume, and the fermentation period was prolonged one week longer. The sialic acid content and bioactivity in vitro of recombinant TNFRp75: Fc were not changed after nutrient compensation and glucose control supply. Nutrient compensating and metabolic control in continual perfusion fermentation could significantly increase the productivity and yield of recombinant TNFRp75:Fc, and thus reduced relative industrialization costs.

Objeetive To explore the influence of extracellular high glucose on the proliferation,migration and biomarkers of corneal limbal stem cells.Methods Establishment of a model of high glucose in cultured human limbal stem cells to observe and investigate the effects of extracellular high glucose on the proliferation and migration of corneal limbal stem cells by immunoflurescence,CCK-8 and Transwell assay,respectively.Totally 16 SPF rats were collected and induced diabetic model by streptozotocin as the high-glucose group,and the normal rats of the same age served as the control group.Corneal epidermises of rats in both groups were scraped to observe the repair of corneal epithelium.And the corneas were treated with HE staining and immunohistochemical staining to detect the expression of biomarkers of corneal limbal stem cells and the modality changes of cells.Results The proliferation rate of human limbal epithelial cells was significantly decreased when exposed to high glucose,and the rate at 24 h,48 h and 72 h was 0.728,0.345 and 0.395,respectively,which was markedly lower than that in the control group,with a significant difference (P ＜ 0.05);meanwhile the cell migration rate of the high-glucose group was 17.6％ at 48 h,which was significantly slower than that of the control group (100％).And the inhibition was accompanied by the decreased expression of β-catenin and vimentin.Furthermore,the expression levels of β-catenin and vimentin mRNA and protein were down-regulated,with abnormal location,in the high-glucose group.And diabetic rats had poor corneal epithelial healing.The epithelial layer became thinner and the structures were disorganized in diabetic rats through HE staining.The immunohistochemical assay revealed the expression of β-catenin and vimentin of cornea limbal stem cells was down-regulated in high-glocose group when compared with the control group.Conclusion High glucose can significantly inhibit the proliferation and migration of cornea limbal stem cells,and its main damage mechanism is correlated with the abnormalities of β-catenin and vimentin.

Objective To investigate the correlation of the evaluation of clinical learning environment (CLE),professional self-concept (PSC),and professional competence (PC) of baccalaureate nursing students when they were in clinical practice.Methods Totals of 194 subjects were recruited by convenient sampling method,data were collected through questionnaire survey.And the questionnaires were included population sociology and general information,evaluation of CLE,PSC,and PC.Results The average scores of CLE evaluation scale,PSC of nurses instrument,and PC scale of baccalaureate nursing students respectively were (3.41 ± 0.60),(2.83 ± 0.34),and (2.90 ± 0.44).Pearson bivariate correlative analysis showed that there were positive correlations between any two of the three scores,the coefficients of correlation r were 0.48 (CLE vs PC,P＜0.01),0.62(CLE vs PSC,P＜0.01),and 0.64 (PC vs PSC,P＜0.01).Conclusions The improvement of CLE is helpful to the development of professional self-concept and professional competence of baccalaureate nursing students.

This study was purposed to investigate whether phenylhexyl isothiocyanate (PHI) can reduce p16 gene methylation level or not. The myeloma U226 cell line was cultured with PHI of 0, 5, 10 micromol/L for 72 hours, then DNA was extracted. Hydrosulfite was used to treat the genome DNA of healthy adult, PCR amplification was carried out by using this DNA as template. The gene chip detecting methylation changes of 3 CpG in promoter region of p16 gene was constructed by designing a pair of probes which contain one methylated and one unmethylated probes. This pair of probes was used to detect 3 consecutive sites of CpG island in p16 gene. The standard curve was constructed by using gene chip after the methylated and unmethylated DNA were mixed at different ratio. Then treated samples of U266 cells were dotted on gene chip, obtained results were compared with standard curve to get the quantitative results. The results indicated that the probes on chip had excellent reproducible ability and precision, the methylation level of p16 gene in U266 cells treated with 0, 5 and 10 micromol/L of PHI was determined as 78.2%, 61.7% and 54.8%, respectively. It is concluded that the PHI can reduce the methylation level of p16 gene in U266 cells.
Cell Line, Tumor ; CpG Islands ; DNA Methylation ; Down-Regulation ; Genes, p16 ; Humans ; Isothiocyanates ; pharmacology ; Oligonucleotide Array Sequence Analysis