Objective To observe the clinical feasibility of the acceptable noise level (ANL) test ,analyze the effect of different test instructions to the result of Mandarin acceptable noise level test .Methods Twelve young peo-ple (24 ears) with normal pure-tone test and acoustic immittance test (threshold≤15 dB HL) were included in this study .They were randomly divided into two groups with 12 ears in each group .Group A received the same test in-struction firstly and then received different instructions depended on the different translation of English instruction by tester .Group B received the reverse test order .The acceptable noise level(ANL) ,most comfortable level(MCL) and back groud noise level(BNL) were analyzed .Results The average value of ANL was 7 .5 ± 6 .61 and 8 .29 ± 6 .54 dB SPL ;the average value of most comfortable level (MCL) was 83 .58 ± 8 .57 and 85 .41 ± 8 .89 dB SPL ;the average value of background noise level (BNL) were 78 .92 ± 10 .56 and 77 .13 ± 7 .91 dB SPL respectivoly .The difference of MCL ,BNL ,ANL in these two groups were not statistically significant (P>0 .05) .Test method 1 re-sulted in great individual differences in ANL ,which it ranged from -6 .4 to 20 .12 dB S/N .While ANL from test method 2 ranged from 2 .50~20 .12 dB S/N .Conclusion Different test instruction do not correlated to the results of acceptable noise level ,testers can translate the English instruction by different ways so that it is convenient to the application and clinical research of ANL test .

Objective To investigate the expression level of LMO1 in gastric cancer tissues and human gastric cancer cell lines,and explore the invasive and metastatic potential of LMO1 gene silencing by small interfering RNA on the human gastric cancer cell line MKN28.Methods Immunohistochemical technique was applied to detect the expression of LOM1 protein in gastric cancer tissues and tumor adjacent tissues of paraffin specimens in 30 cases.The expression levels of LMO1 in human gastric cancer cell lines AGS,BGC-823,SGC-7901,MKN28 and human gastric mucosal epithelial cells GES were detected by realtime-PCR and Western blot.Using LipofectamineTM 2000,LMO1 siRNA was transfected into MKN28 cellsin vitro (siRNA transfect group).Negative control was established.Real time-PCR and Western blot was used to examine the difference of LMO1 expression.Transwell assays were performed to identify the differences and changes of invasive and metastatic ability in gastric cancer cell line MKN28.Western blot was used to examine the expression levels of E-cadherin,MMP-9 and VEGF.Results Positive rate of LOM1 protein in gastric cancer tissues(77％)was higher than that in tumor adjacent tissues (17％) (x2 =21.70,P ＜ 0.01).Positive rate of Vavl protein was higher in lymphatic metastasis group than in non-lymphatic metastasis group(x2 =5.83,P =0.02).Compared with GES,the expression level of LMO1 increased significantly in gastric cancer cell lines,especially in MKN28 (P ＜ 0.01).The expression levels of LMO1 mRNA and protein in LMO1-siRNA transfected MKN28 cells were lower than the matched negative control cells (P ＜0.01).The invasive and metastatic potentials of LMO1-siRNA transfected MKN28 cellssignificantly decreased (t =-11.53,P ＜0.01;t =-10.68,P ＜0.01).The expression levels of E-cadherin were higher than the matched negative control cells;and MMP-9,VEGF protein in LMO1-siRNA transfected MKN28 cells were lower than the matched negative control cells (P ＜ 0.01).Conclusions LMO1 has higher expression level in gastric cancer tissues and some gastric cancer cell lines,and down-regulation of LMO1 can inhibit the invasion and metastasis ability of gastric cancer.

Objective:To evaluate the relationship between changes in Golgi apparatus morphological structure and endotoxin-induced acute lung injury (ALI) in mice.Methods:Twenty healthy male C57BL/6J mice, weighing 18-20 g, aged 6-8 weeks, were divided into 2 groups ( n=10 each) using a random number table method: sham operation group (group Sham) and endotoxin-induced ALI group (group ALI). Lipopolysaccharide (LPS) 10 mg/kg was injected intravenously in group ALI, while the equal volume of normal saline 0.5 ml was given instead in group Sham.The animals were sacrificed at 12 h after LPS injection and the lung tissues were taken for detection of the content of reactive oxygen species (ROS) and wet to dry weight ratio (W/D ratio), for observation of the pathological changes (using HE staining) and Golgi apparatus morphological structure (with a transmission electron microscope) and for determination of expression of Golgi matrix protein 130 (GM130), Golgin97 and mannosidase alpha class II member 1 (MAN2A1) and its mRNA (by Western blot and quantitative polymerase chain reaction). Results:Compared with group Sham, ROS content and the W/D ratio in lung tissues were significantly increased, GM130, MAN2A1, Golgin97 protein and its mRNA expression were down-regulate ( P<0.01), the pathological changes of lung tissues were accentuated, the Golgi cisternae was swollen, and Golgi fragments were dispersed in the cytoplasm in group ALI. Conclusion:The mechanism of endotoxin-induced ALI may be related to the changes in Golgi apparatus morphological structure.

BACKGROUND: The investigation of culturing, passaging and establishing human limbal stem cells can strengthen the recognition of the stem cells and provide the enough cellular reserve for the basic and clinical research of limbal stem cell transplantation.OBJECTIVE: To explore a method of pessaging and establishing cell line of human limbal stem cells cultured in vitro.DESIGN: Randomized controlled observation.SETTING: Gannan Medical College.MATERIALS: The experiment was performed at Scientific Center of Gannan Medical College and the National Key Laboratory of Ophthalmology Hospital Affiliated to Sun Yat-Sen University from June 2003 to April 2004. Fresh human limbus corneae were isolated from two healthy donors. Procedures were performed according to the informed consent of the donors. Main reagents contained RPMI-1640 (Sigma R8755, containing L-glutamine) and 200 g/L fetal calf serum (FCS) (Gibco 16140-071). DMEM medium, chondroitin sulfatase and human epidermal growth factor (hEGF) were purchased from Sigma Co. USA; HEPES and DMSO were bought from Gibco, USA; 100% glycerinum was purchased from Yunjia Huangpu Pharmaceutical Product Limited Company, PRC; glutaraldehyde was bought from E.Merk, Germany; Alcohol, chlorhydric acid, acetone and methyl aldehyde were purchased from Beijing Chemical Agent Company, PRC; 0.25% parenzyme was bought from Shanghai Xinhua Pharmaceutical Factory, PRC.Above-mentioned reagents were analytical pure grade.METHODS: After digestion, human limbal tissues in limbal basilar part with an abundant pigment were cultured in the culture flask containing RPMI-1640 and 200 g/L FCS and in culture dish containing amniotic extracellular matrix (AECM) as the cultural supporter. Primary and passage cells were observed under light microscope and scanning electron microscope (SEM). The revival ratio of stem cell refrigeration of every generation was calculated by the trypanblau exclusion experiment.MAIN OUTCOME MEASURES: ① Observational results of limbal stem cells during the primary culture and serial subcultivation in vitro, and ② revival ratio of stem cell refrigeration.RESULTS: ①Findings of primary culture: Most limbal stem cells in the culture flask had the adherence and were arrayed uniformly sparsely to form monolayer and adhered to the bottom of culture flask under the inverted phase contrast microscope after 1-day culture. ② Findings of serial subcultivation: After human epidermal growth factor (hEGF) was added into the second passage, cells were scattered into the monolayer and adhered to grow quickly.Morphological variability of all the cells increased obviously when passage the 30th generation. The cellular volume was obviously increasing, and the round or irregular round cells gathered together. The 33rd generation human limbal stem cells still could vigorously differentiate, proliferate and grow in ACEM. ③ The revival ratio of stem cell refrigeration was 82.2%.CONCLUSION: The human limbal stem cell lines were preliminarily established by culturing and freezing the cells of 33 generations in vitro. The human limbal stem cell lines preferred to grow in the culture dish containing AECM as the cultural supporter.

Objective To investigate the expression and clinical significance of neutrophil S100A8/A9 in induced sputum in children with bronchial asthma. Methods A total of 108 cases of bronchial asthma patients in the FourthAffiliated Hospital of Nanchang University were involved in the study form October 2014 to October 2015. According to the severity of the disease, the patients were divided into mild group (n=40), moderate group (n=36) and severe group (n=32). Twenty health children were taken as control group at the same period. All the patients were treated with budesonide aerosol for three months, and the control group was received aerosol inhalation for normal saline (NS). The ratio of forced expiratory volume in one second and forced vital capacity (FEV1/FVC, FEV1%) were used to evaluate the pulmonary function in two groups. The asthma control questionnaire (AcQ-5) score was used to estimate the asthma control effects. The expression level of neutrophil S100A8/A9 mRNA in induced sputum was detected by real-time PCR. The correlation of S100A8/A9 mRNA, AcQ-5 score and FEV1%was analyzed. Results Before the treatment, the FEV1%decreased, while the AcQ-5 score and express level of S100A8/A9 mRNA significantly increased with the severity of disease (all P<0.01). Three months after treatment, asthma was completely controlled in 60 patients, partial controlled in 31 cases and uncontrolled in 17 cases. With the improvement of the therapeutic efficacy, the FEV1%significantly decreased, while the express level of S100A8/A9 mRNA significantly increased (all P < 0.01). The express level of S100A8/A9 mRNA in induced sputum neutrophils was negatively correlated with FEV1%(r=-0.327 and-0.406 respectively, P<0.05), which was positively correlated with ACQ-5 score (r=0.704 and 0.817, P<0.05). Conclusion The level of S100A8/A9 expression in induced sputum neutrophil is positively correlated with the severity of asthma, which can be used as clinical indicators of the severity and the efficacy of asthma.

Serum high sensitivity C-reactive protein(hs-CRP)and glycosylated hemoglobin(HbA1c )were detected in 89 type 2 diabetes patients with chronic periodontitis.The patients were divided into HbA1c≥ 7.0% and <7.0% groups according HbA1c levels.Serum hs-CRP,probing depth(PD),attachment loss(AL)in HbA1c ≥ 7.0% group were all significantly higher than those in <7.0% group (P <0.01 ),and serum hs-CRP was positively correlation with PD and AL in all patients.In patients with type 2 diabetes,high HbA1c and chron-ic periodontitis serum hs-CRP is positively correlation with periodontal disease.

Objective:To evaluate the effect of a hand hygiene culture program on hand hygiene compliance in a three grade A hospital .Methods:Direct observation methods were used to assess the hand hygiene compliance and hand hygiene quality .Results:After the hand hygiene culture program , the hand hygiene compliance rate and the hand hygiene accuracy rate of doctors raised from 30 .2%and 66 .7%to 65 .3%and 85 .8%.Those of the nur-ses raised from 52.2%and 80.0%to 87.6%and 93.3%.Conclusion:The hand hygiene program increased the hand hygiene compliance and reducing thd risk of hospital infection occurred .

Objective: To learn the mutation types and hearing screening results in local newborns of Zhoushan,in order to provide evidence for prevention and early detection of deafness. Methods: The newborns in Zhoushan Maternal and Child Health Hospital from August 2015 to May 2018 were recruited and detected by matrix-assisted laser desorption ionization time of flight mass spectrometry（MALDI-TOF-MS）for twenty-two mutation sites of GJB2,SLC26A,GJB3 and 12SrRNA genes. The results of genotyping and hearing screening were analyzed and the hearing condition of abnormal newborns was followed up. Results: Among 4 029 newborns,180（4.47%）newborns were identified to carry mutations,including 94 males（4.66%）and 86 females （4.28%）. There was no statistically significant difference in the rate of carrying mutations between male and female infants （P>0.05）. Totally 135 （3.35%）newborns failed in primary hearing screening,13（9.63%）of whom carried the deafness genes;3 894（96.65%）newborns passed,167（4.29%）of whom carried the deafness gene. There was statistically significant difference in the the rate of carrying mutations between newborns who passed and failed in primary hearing screening （P<0.05）. Eleven newborns were diagnosed with hearing loss,with a rate of 2.73‰. Among 180 mutations identified,there were 91 GJB2 mutations（2.26%）,57 SLC26A4 mutations（1.41%）,14 GJB3 mutations （0.35%）,15 mtDNA 12SrRNA mutations （0.37%）and 3 with mutations of two genes （0.07%）. Sixteen mutation sites （184 cases）were found,and the detection rate was 4.57%. Conclusion The rate of carrying deafness genes in Zhoushan newborns was 4.47%. The deafness genes found were mainly GJB2 and SLC26A4,the carrying rate of mtDNA 12SrRNA gene mutation was also high.

Objective TGF-β1 can promote EMT,then strengthen the invasion and metastasis ability of cancer ceils.However,the mechanism for TGF-β1 in gastric cancer still keeps unclear.Aim of this study was to investigate the expression of epithelial to mesenchymal transition (EMT)marker,LMO1 and metastasis related genes on the human gastric cancer cell cell line MKN28 treated with TGF-β1,and test whether down-regulate LMO1 expression can affect the pro-EMT and pro-metastatic roles of TGF-β1 in MKN28 cells.Methods Primary human gastric cancer cell line MKN28 was cultured in vitro.Cells were treated with TGF-β1 to induce cells to undergone EMT.Cells were divided into four groups:control group (5 ％ BSA),TGF-β1 induced group (10 μg/L),negative transfect group (TGF-β1 +negative transfect siRNA),and LMO1-siRNA transfect group (TGF-β1+ LMO1-siRNA).Real time-PCR and Western blot was used to examine the difference of EMT marker (E-cadherin and N-cadherin),LMO1 and metastasis related genes (MMP-9 and VEGF)expression.Transwell assays were performed to identify the differences and changes of invasive and metastatic ability in gastric cancer cell line MKN28.Western blot was used to examine the expression levels of MMP-9 and VEGF.Results TGF-β1 stimulation induced classical EMT morphological change,as was confirmed by E-cadherin decrease and N-cadherin,LMO1,MMP-9,VEGF increase (P＜0.01).Accompanied with the EMT,cell invasion and migration ability was markedly increased (P＜0.01).However,Down-regulation of LMO1 expression reversed the pro-migratory effect of TGF-β1 to a great degree (P＜0.01).Conclusion LMO1 played a central role in coordinating TGF-β1 induced EMT and pro-migratory effects in gastric cancer MKN28 cells.Using siRNA to downregulate the expression of LMO1 can inhibit the invasion and metastasis ability of gastric cancer MKN28 cells.