Objectives: . Reactive oxygen species in the stria vascularis (SV) of the cochlea may be involved in the pathogenesis of sensorineural hearing loss. However, the effects of oxidative stress on SV endothelial cells (SV-ECs) remain largely unknown, and no feasible in vitro cell culture model exists for the functional study of SV-ECs. Methods: . We isolated primary SV-ECs from the SV of neonatal mice. The apoptosis-reducing effects of fibronectin in SV-ECs cultured with serum-free medium were determined using β-galactosidase staining and flow cytometry. SV-ECs incubated in serum-free medium were treated with various H2O2 concentrations to evaluate the effects of H2O2 on their viability. The secretome of SV-ECs treated with or without H2O2 (100 μM or 500 μM) was analyzed using high-resolution mass spectrometry. The function of the SV-EC secretome was evaluated by a macrophage assay. Results: . We successfully isolated and characterized the SV-ECs. Treatment with H2O2 at concentrations up to 500 μM for 2 hours and further incubation with serum-free medium in plates precoated with fibronectin showed no significant effect on apoptosis. Compared to the control SV-ECs, the amount of differential proteins in the secretome of SV-ECs stimulated with 500 μM H2O2 was much higher than in those treated with 100 μM H2O2. Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analyses suggested that the proteins differentially expressed in SV-ECs treated with 500 μM H2O2 were involved in the regulation of multiple signaling pathways and cellular processes. The secretome of H2O2-stimulated SV-ECs exhibited significant pro-inflammatory effects on macrophages. Conclusion . We successfully established an in vitro serum-free culture method, identified the differential proteins released by oxidative stress-induced ECs and their functions, and revealed the pro-inflammatory effects of the secretome of H2O2-stimulated SV-ECs. Therefore, SV-ECs might elicit immunoregulatory effects on bystander cells in the microenvironment of oxidative stress-induced cochlea, especially cochlear macrophages.

Additional sex combs-like 1 (ASXL1) interacts with BRCA1-associated protein 1 (BAP1) deubiquitinase to oppose the polycomb repressive complex 1 (PRC1)-mediated histone H2A ubiquitylation. Germline BAP1 mutations are found in a spectrum of human malignancies, while ASXL1 mutations recurrently occur in myeloid neoplasm and are associated with poor prognosis. Nearly all ASXL1 mutations are heterozygous frameshift or nonsense mutations in the middle or to a less extent the C-terminal region, resulting in the production of C-terminally truncated mutant ASXL1 proteins. How ASXL1 regulates specific target genes and how the C-terminal truncation of ASXL1 promotes leukemogenesis are unclear. Here, we report that ASXL1 interacts with forkhead transcription factors FOXK1 and FOXK2 to regulate a subset of FOXK1/K2 target genes. We show that the C-terminally truncated mutant ASXL1 proteins are expressed at much higher levels than the wild-type protein in ASXL1 heterozygous leukemia cells, and lose the ability to interact with FOXK1/K2. Specific deletion of the mutant allele eliminates the expression of C-terminally truncated ASXL1 and increases the association of wild-type ASXL1 with BAP1, thereby restoring the expression of BAP1-ASXL1-FOXK1/K2 target genes, particularly those involved in glucose metabolism, oxygen sensing, and JAK-STAT3 signaling pathways. In addition to FOXK1/K2, we also identify other DNA-binding transcription regulators including transcription factors (TFs) which interact with wild-type ASXL1, but not C-terminally truncated mutant. Our results suggest that ASXL1 mutations result in neomorphic alleles that contribute to leukemogenesis at least in part through dominantly inhibiting the wild-type ASXL1 from interacting with BAP1 and thereby impairing the function of ASXL1-BAP1-TF in regulating target genes and leukemia cell growth.

[Objective] To evaluate the efficacy of sound therapy and provide a kind of treatment for tinnitus patients with normal audiogram.[Methods] From the database of the Audiometric Center of the First Affiliated Hospital,Sun Yat-sen University,we identified 46 outpatients with normal hearing thresholds in the conventional pure-tone audiometry suffering subjective tinnitus over 3 months.The clinical information were collected with respect of age,gender,tinnitus severity,pitch,laterality and duration,comorbid symptoms.All the patients received the tinnitus test,including loudness and frequency of the tinnitus and residual inhibition,Tinnitus Handicap Inventory (THI) and Visual Analogue Scale (VAS) scores pre-and post-complex sound therapy.The complex sound were remixed with natural sounds and music materials,while the masking frequency of natural sounds is consistent with the tinnitus frequency of patients and the main frequency of music materials covers the tinnitus frequency.[Results] 32 of the 46 patients showed residual inhibition (RI) positive.The average THI score before treatment was 33.38±16.23 and the VAS score was 4.38±1.62.The THI score decreased to 21.75±11.67 and the VAS score was 2.97±1.06 after sound therapy.The rest 14 patients were residual inhibition (RI) negative.The THI score were 29.86±20.15 and 23.43±16.29 pre-and post-complex sound therapy,while the VAS score were 3.93±1.69 and 3.36±1.2.The THI and VAS scores were significantly decreased after the treatment in both RI positive and negative patients.The RI positive group showed better results than RI negative group.[Conclusions] Both RI positive and negative tinnitus with normal audiometry shown adaptation with complex sound therapy,RI positive patients acquired more benefits from tailor-made complex sound therapy.

Objective The expression level of the Ki-67 protein is a marker of the poor prognosis in non-small-cell lung cancer. This study explored the correlation of the Ki-67 expression level with the clinical efficacy in the treatment of advanced lung adenocarcinoma.Methods From January 2015 to December 2015, 92 patients with stage Ⅳ lung adenocarcinoma were treated in the Department of Respiratory and Critical Care Medicine of Nanjing General Hospital, 65 with oral gefitinib for EGFR mutation-positive or crizotinib for ALK positive (the targeted therapy \[TT\] group) and the other 27 with pemetrexed + cisplatin or nedaplatin (the chemotherapy \[CT\] group). The expression of Ki-67 was determined by immunohistochemistry, its relationship with the clinicopathological features and therapeutic effects was analyzed, and the progression-free survival (PFS) was calculated.Results Ki-67 was expressed lowly in 43 (46.7%) and highly in 49 (53.3%) of the cases. The median PFS after treatment was significantly longer in the patients with a low than in those with a high Ki-67 expression in the TT group (10 vs 7 mo, P<0.05) and in the CT group as well (8 vs 7 mo, P<0.05). Ki-67 (HR=1.011, 95% CI: 1.000-1.023) and ALK (HR=0.325, 95% CI: 0.112-0.942) were shown to be predictive factors of poor PFS.Conclusion The expression level of Ki-67 affects the effect of clinical treatment, especially that of the first-line targeted therapy, on advanced lung adenocarcinoma.

BACKGROUNDNowadays, social media tools such as short message service, Twitter, video, and web-based systems are more and more used in clinical follow-up, making clinical follow-up much more time- and cost-effective than ever before. However, as the most popular social media in China, little is known about the utility of smartphone WeChat application in follow-up. In this study, we aimed to investigate the feasibility and superiority of WeChat application in clinical follow-up.

METHODSA total of 108 patients diagnosed with head and neck tumor were randomized to WeChat follow-up (WFU) group or telephone follow-up (TFU) group for 6-month follow-up. The follow-ups were delivered by WeChat or telephone at 2 weeks, 1, 2, 3, and 6 months to the patients after being discharged. The study measurements were time consumption for follow-up delivery, total economic cost, lost-to-follow-up rate, and overall satisfaction for the follow-up method.

RESULTSTime consumption in WFU group for each patient (23.36 ± 6.16 min) was significantly shorter than that in TFU group (42.89 ± 7.15 min) (P < 0.001); total economic cost in WFU group (RMB 90 Yuan) was much lower than that in TFU group (RMB 196 Yuan). Lost-to-follow-up rate in the WFU group was 7.02% (4/57) compared with TFU group, 9.80% (5/51), while no significance was observed (95% confidence interval [CI]: 0.176-2.740; P = 0.732). The overall satisfaction rate in WFU group was 94.34% (50/53) compared with 80.43% (37/46) in TFU group (95% CI: 0.057-0.067; P = 0.034).

CONCLUSIONSThe smartphone WeChat application was found to be a viable option for follow-up in discharged patients with head and neck tumors. WFU was time-effective, cost-effective, and convenient in communication. This doctor-led follow-up model has the potential to establish a good physician-patient relationship by enhancing dynamic communications and providing individual health instructions.

TRIAL REGISTRATIONChinese Clinical Trial Registry, ChiCTR-IOR-15007498; http://www.chictr.org.cn/ showproj.aspx?proj=12613.

Adult ; Aftercare ; economics ; methods ; Aged ; Female ; Head and Neck Neoplasms ; Humans ; Male ; Middle Aged ; Patient Discharge ; economics ; statistics & numerical data ; Smartphone ; Social Media ; Telephone ; Young Adult

OBJECTIVETo improve the reception and recruitment of sperm donors in sperm banks in China, and solve the problem of insufficiency in sperm donation.

METHODSWe reviewed the recruitment of 1 145 men for sperm donation in the Human Sperm Bank of Hubei Province from September 2011 to April 2012, analyzed the reasons for those not included, and interviewed those included but unwilling to donate sperm.

RESULTSAmong the 1 145 recruits, 551 (48.12%) were students and 594 (51.88%) were other individuals. After the first semen screening, 503 (43.93%) quitted, including 202 students (36.66% of the students recruited) and 301 others (50.67% of the other individuals recruited). After the second semen screening, 432 (37.73%) were excluded, and another 45 (3.93%) excluded after laboratory examination, including 16 cases of mycoplasma positive. Totally, 165 recruits (14.41%) passed the semen screening and laboratory examination, but only 144 of them (87.27%) completed, while the other 21 (12.73%) failed to complete the whole donation process.

CONCLUSIONLow rates of screening qualification and donation process completion are common problems in human sperm banks. The rate of qualified sperm donors can be increased and the operational cost of the human sperm bank can be reduced by enabling the recruits to accomplish the whole donation process. Explanation at the reception, later interview with the recruits, and donors' trust in the sperm bank play important roles in raising the completion rate of sperm donation process.

Adult ; Humans ; Male ; Semen ; Sperm Banks ; Tissue Donors ; psychology ; Tissue and Organ Procurement

OBJECTIVETo explore the effects of bisphenol A (BPA) exposure on toxicity characteristic and OCT4 and SOX2 gene expression of mouse embryonic stem cells (mESC).

METHODSmESC were cultured, and treated with the doses of 10(-8), 10(-7), 10(-6), 10(-5), 10(-4) mol/L respectively of BPA and DMSO (the solvent control group)for 24 hours, and three groups of cells were treated with the same method. The morphological changes of mESC in the control and exposure groups were observed through an inverted microscope. Cell counting kit 8 (CCK8) was used to detect the effects of BPA on proliferation of mESC, and based on the results, the half inhibitory concentration (IC50) was calculated. Real-time fluorescent quantitative polymerase chain reaction (RT-QPCR) and western blotting were used to detect the expression of OCT4 and SOX2.

RESULTSBPA had certain toxicity on mESC, the treatment of BPA significantly increased cell toxicity in a concentration-dependent manner, and the IC50 was 4.3×10(-4) mol/L, combined with the BPA exposure concentration of the environment and the related literature, eventually taking the five concentrations of 10(-8), 10(-7), 10(-6), 10(-5), 10(-4) mol/L as the experimental groups. The mESC morphology were effected after the treatment of BPA for 24 h, compared with the control group, the number of cells decreased, appearing some floating cells, and the cell cloning became irregular and differentiation in the higher concentration groups. The OCT4 mRNA expression level in the 10(-7) mol/L (1.146 ± 0.087), 10(-6) mol/L (1.156 ± 0.030), 10(-5) mol/L (1.158 ± 0.103) and the 10(-4) mol/L (1.374 ± 0.053) dose group were all significantly higher than the control group (1.000 ± 0.000) (t values were -2.384, -2.953, -3.203, -4.021 respectively, P value all < 0.05). Meanwhile, the SOX2 mRNA expression level in the 10(-4) mol/L (1.113 ± 0.052) were higher than the control group (1.000 ± 0.000) (t value was -2.765, P value < 0.05). Moreover, the OCT4 protein expression level in the 10(-5) mol/L (1.360 ± 0.168) and 10(-4) mol/L (1.602 ± 0.151) were all significantly higher than the control group (1.000 ± 0.000) (t values were -3.538, -4.002 respectively, P value all < 0.05), while no obvious change of the SOX2 protein expression level was detected in all treated groups.

CONCLUSIONBPA in a certain dose range could upregulate the expression of OCT4 gene in mouse embryonic stem cells while had no significant effect on the expression of SOX2 gene.

Animals ; Benzhydryl Compounds ; toxicity ; Cells, Cultured ; Embryonic Stem Cells ; drug effects ; metabolism ; Gene Expression ; Mice ; Octamer Transcription Factor-3 ; genetics ; Phenols ; toxicity ; SOXB1 Transcription Factors ; genetics ; Signal Transduction ; drug effects

The aim of study was to investigate the effect of acute lymphoblastic leukemia (ALL) children bone marrow mesenchymal stem cells (MSC) on resistance of K562/A02 cells and its mechanism. MSC obtained from bone marrow of AL children were cultured and identified. The co-culture of MSC and K562/A02 and the culture of K562/A02 cell suspension alone was performed, of which 2 kinds of cells were treated with same concentration of adriamycin (ADM), and the rate of apoptosis was detected by flow cytometry, bcl-2 and bax of K562/A02 were detected by RT-PCR, while mdr1 gene level was detected by FQ-PCR. The results indicated that the MSC separation and proliferation were viable and steady. The apoptosis rate of the K562/A02 cells co-cultured with MSC was 1.97 ± 0.11%, while apoptosis rate of the K562/A02 cells cultured alone was 8.38 ± 0.29%, there was significant difference (p < 0.05). As compared with the K562/A02 cells cultured alone, the bcl-2 gene expression in K562/A02 cells co-cultured with MSC obviously increased; ratio of bcl-2/bax was obviously enhanced. The mdr1 gene level in K562/A02 co-cultured with MSC was no statistical different from K562/A02 cultured alone (p > 0.05), which suggested that adhesion co-cultured with MSC did not induce mdr1 expression higher than the culture of suspension. It is concluded that the MSC of ALL children can escape the leukemia cells from proapoptotic effect of drugs, the resistance of K562/A02 to ADM may be involved in enhancement of bcl-2 gene expression of K562/A02 cells co-cultured with MSC, but not in relation to mdr1 gene in K562/A02 cells themselves.
ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Bone Marrow Cells ; drug effects ; Child ; Child, Preschool ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Female ; Gene Expression Regulation, Leukemic ; Humans ; K562 Cells ; Male ; Mesenchymal Stromal Cells ; drug effects ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; bcl-2-Associated X Protein ; genetics

Objective To study the aerodynamics of the normal human nasal cavity under different ambient temperatures.Methods Based on CT scanning,a model of a healthy adult's nasal cavity was established using computational fluid dynamics software from Fluent.Airflow in this model was simulated and calculated at ambient temperatures of 0℃,24℃,and 37℃ during periodic breathing.Results Ambient temperature only had an impact on the temperature in the nasal cavity during the inspiratory phase,and the temperature distribution was not symmetrical in the inspiratory acceleration and deceleration phases.The ambient temperature significantly affected airflow speed in main nasal passages during the inspiratory process,but had little impact on flow status (proportion and streamline of airflow in different nasal passages).Temperature differences increased the irregular air movement within sinuses.The anterior nasal segment,including the area between the valve and the head of the middle turbinate,was the most effective part of the nasal airway in heating the ambient air.Conclusions Our findings describe the effects of ambient temperature on airflow parameters in the nasal cavity within a single respiratory cycle.This data is more comprehensively and accurately to determine the relationship between nasal cavity aerodynamics and physiological functions.

BACKGROUND AND OBJECTIVERadiation usually results in paranasal sinusitis in patients with nasopharyngeal carcinoma (NPC), which influences patients' quality of life. This study aimed to determine the relationships between dose distribution in the nasal cavity and nasal mucous injury in patients with NPC treated by intensity-modulated radiation therapy (IMRT), and to find the tolerable radiation dose for the nasal mucous.

METHODSSixty-six patients with NPC treated by IMRT between October 2006 and November 2008 were enrolled. The irradiation dose in the nasal cavity was determined by the computer with the IMRT work platform. Mucociliary transport rate (MTR) was detected by modified saccharine test before IMRT, at the end of IMRT, and at 3, 6, and 12 months after IMRT.

RESULTSThe data were available for 129 nasal cavities. The cavities receiving a mean dose below or equal to 37 Gy showed substantial preservation of nasal mucous after IMRT. The MRT decreased to (62.82 ± 38.59)%, (56.78 ± 37.79)%, (64.05 ± 39.37)%, and (71.13 ± 39.55)% of pre-IMRT value at 4 time points after IMRT, with significant differences among the data (P < 0.05). In contrast, when the cavities received a mean dose higher than 37 Gy, no significant differences in MTR among the time points were observed. At 3 months after IMRT, the MTR was the lowest (38.27% of pre-RT value).

CONCLUSIONSA mean radiation dose of ≤ 37 Gy for the nasal cavity is an optimal dose to protect the nasal cavity function.

Adult ; Carcinoma, Squamous Cell ; physiopathology ; radiotherapy ; Female ; Humans ; Male ; Mucociliary Clearance ; radiation effects ; Nasal Cavity ; radiation effects ; Nasopharyngeal Neoplasms ; physiopathology ; radiotherapy ; Quality Control ; Radiotherapy Dosage ; Radiotherapy, Intensity-Modulated