Isolation and Culture of Primary Cochlear Hair Cells from Neonatal Mice.

Cochlear hair cells are the sensory receptors of the auditory system. These cells are located in the organ of Corti, the sensory organ responsible for hearing, within the osseous labyrinth of the inner ear. Cochlear hair cells consist of two anatomically and functionally distinct types: outer and inner hair cells. Damage to either of them results in hearing loss. Notably, as inner hair cells cannot regenerate, and damage to them is permanent. Hence, in vitro cultivation of primary hair cells is indispensable for investigating the protective or regenerative effects of cochlear hair cells. This study aimed to discover a method for isolating and cultivating mouse hair cells. After manual removal of the cochlear lateral wall, the auditory epithelium was meticulously dissected from the cochlear modiolus under a microscope, incubated in a mixture consisting of 0.25% trypsin-EDTA for 10 min at 37 °C, and gently suspended in culture medium using a 200 µL pipette tip. The cell suspension was passed through a cell filter, the filtrate was centrifuged, and cells were cultured in 24-well plates. Hair cells were identified based on their capacity to express a mechanotransduction complex, myosin-VIIa, which is involved in motor tensions, and via selective labeling of F-actin using phalloidin. Cells reached >90% confluence after 4 d in culture. This method can enhance our understanding of the biological characteristics of in vitro cultured hair cells and demonstrate the efficiency of cochlear hair cell cultures, establishing a solid methodological foundation for further auditory research.

[MRI Findings of the Uterine Tumors Resembling Ovarian Sex Cord Tumors:Report of Two Cases].

Uterine tumors resembling ovarian sex cord tumors are rarely reported with limited imaging findings.The current study reported two case of uterine tumors resembling ovarian sex cord tumors and described the detailed MRI findings,which would provide valuable imaging evidence for the diagnosis of such tumors.

[Effect of Xuanfu Daizhe decoction on stemness of esophageal cancer cells].

Objective: To investigate the effect of Xuanfu Daizhe decoction on the stemness of esophageal cancer cells. Methods: The BALB/c nude mice were randomly divided into the control group and experimental group, 5 mice in each group, which were continuously administered with normal saline and Xuanfu Daizhe decoction (9.89 g/kg) by gastrogavage, respectively. Human esophageal carcinoma cells ECA-109 (5×106) were subcutaneously injected into the mice on the 8th day. Tumor volume was measured twice a week. The mice were sacrificed 4 weeks after injection, and the tumor tissue and mouse serum were collected. The expressions of the major stemness-regulating transcription factors, i.e., NANOG, OCT4 and SOX2, were detected by RT-qPCR, Western Blot and immunohistochemistry. ECA-109 cells were treated with 10% fetal bovine serum and serum from the above two groups of mice for 48 hours respectively, and three replicate wells were set in each group, and the expressions of NANOG, OCT4, SOX2 and the levels of AKT and p-AKT were detected by RT-qPCR and Western Blot, respectively. ALDH activity in tumor cells was detected by flow cytometry; the number of spheroids of tumor cells was detected by the spheroidization experiment. Results: Compared with the control group, the growth and size of esophageal cancer tumors were significantly inhibited by Xuanfu Daizhe Decoction; the expressions of NANOG, OCT4, SOX2, the ALDH activity, the number of spheroids, and the levels of AKT and phosphorylated AKT (p-AKT) in esophageal cancer cells were significantly reduced by Xuanfu Daizhe Decoction both in vivo and in vitro. Conclusion: Xuanfu Daizhe Decoction inhibits the stemness of esophageal cancer cells, it may be a potentially effective drug for the treatment of esophageal cancer and provides a theoretical basis for the exploration of new effective drugs for the treatment of esophageal cancer.

[Two Cases of Hypovascular Pituitary Metastases and Literature Review].

Pituitary metastasis,a rare kind of intracranial malignant tumor,is characterized by metastasis from all parts of the body to the pituitary.The common tumors metastatic to pituitary are abundant with blood supply,located in the posterior pituitary lobe and/or the pituitary stalk.The lesion shows infiltrative growth,and mainly demonstrates low signal on T1 weighted imaging and high signal on T2 weighted imaging.It is usually enhanced significantly after iodinated contrast administration.The metastatic tumor with poor blood supply is similar to pituitary macroadenoma on magnetic resonance image,which makes it difficult to be differentiated.We reported two cases of pathologically diagnosed pituitary metastasis with poor blood supply and reviewed related papers,aiming to provide the imaging differentiation points of hypovascular pituitary metastasis.

Effect of MR Field Strength on the Texture Features of Cerebral T2-FLAIR Images: A Pilot Study.

Objective To investigate effect of MR field strength on texture features of cerebral T2 fluid attenuated inversion recovery (T2-FLAIR) images. Methods We acquired cerebral 3D T2-FLAIR images of thirty patients who were diagnosed with ischemic white matter lesion (WML) with MR-1.5T and MR-3.0T scanners. Histogram texture features which included mean signal intensity (Mean), Skewness and Kurtosis, and gray level co-occurrence matrix (GLCM) texture features which included angular second moment (ASM), Contrast, Correlation, Inverse difference moment (IDM) and Entropy, of regions of interest located in the area of WML and normal white matter (NWM) were measured by ImageJ software. The texture parameters acquired with MR-1.5T scanning were compared with MR-3.0T scanning. Results The Mean of both WML and NWM obtained with MR-1.5T scanning was significantly lower than that acquired with MR-3.0T (P<0.001), while Skewness and Kurtosis between MR-1.5T and MR-3.0T scanning showed no significant difference (P>0.05). ASM, Correlation and IDM of both WML and NWM acquired with MR-1.5T revealed significantly lower values than those with MR-3.0T (P<0.001), while Contrast and Entropy acquired with MR-1.5T showed significantly higher values than those with MR-3.0T (P<0.001). Conclusion MR field strength showed no significant effect on histogram textures, while had significant effect on GLCM texture features of cerebral T2-FLAIR images, which indicated that it should be cautious to explain the texture results acquired based on the different MR field strength.

[Clinical Efficacy and Prognostic Factors of Newly Diagnosed Children[JP] with Burkitt Lymphoma Treated by High-Dose and Short-Course Modified LMB Regimen ± Rituximab].

OBJECTIVE: To investigate the clinical efficacy and prognostic factors of high-dose and short-course modified LMB regimen ± Rituximab in the treatment of newly diagnosed children with Burkitt lymphoma, soas to provide more reference for clinical diagnosis and treatment with follow-up. METHODS: 91 newly diagnosed children with Burkitt lymphoma treated in our hospital from January 2007 to August 2016 were chosen. High-dose and short-course modified LMB regimen were used to treat children at different risk levels- Rituximab were added in treatment of high-risk group. The clinical characteristics and efficacy of the treatment were analyzed, and the related prognostic factors were evaluated. RESULTS: The overall survival rate and event-free survival rate in 5-year with follow-up of 91 children were separately（89.27±2.69）%， （87.16±2.30）%; the event-free survival rate of patients in 5-year with follow-up in low-risk, moderate-risk and high-risk group were separately 100%，（94.51±2.97）%，（84.60±3.40）%. The event-free survival rate in 5-year with follow-up of high-risk group were significantly lower than that of moderate-risk group（P<0.05）. Univariate analysis showed that the combination of maxillofacial and central nervous system invasion, LDH >1000 U/L, proportion of bone marrow tumor cell >25%, organ involvement number >4, St. Jude stage for IV stage, early chemotherapy insensitivity and tumor lesion in mid-term evaluation were risk factor for poor prognosis in newly diagnosed children with Burkitt lymphoma（P<0.05）.Multivariate analysis showed that the combination of maxillofacial and central nervous system invasion, early chemotherapy insensitivity and tumor lesion in mid-term evaluation were the independent risk factor for poor prognosis of children with Burkitt lymphoma（P<0.05）. The EFS rate in 5-year with follow-up of high-risk children treated with chemotherapy+rituximab was significantly higher than that of chemotherapy alone（P<0.05）. CONCLUSION: High-dose and short-course modified LMB regimen in the treatment of newly diagnosed children with Burkitt lymphoma shows satisfactory clinical efficacy. The children with central nervous system involvement, early chemotherapy insensitivity and tumor lesion in mid-term evaluation show the worse prognosis, while Rituximab added is more helpful to improve the long-term prognosis for high-risk children with Burkitt lymphoma.

[Hypovascular Pituitary Metastasis from Poorly Differentiated Lung Adenocarcinoma:Report of One Case].

Pituitary metastasis is a rare intracranial malignant tumor originated from other malignant tumors at other parts of the body.Pituitary metastases are typically hypervascular,while hypovascular pituitary metastasis is rare.Radiologically,hypovascular pituitary metastasis is difficult to be distinguished from pituitary adenoma.This article reports one case of pathologically confirmed hypovascular pituitary metastasis,with the main diagnostic clues including history of malignant tumor,presence of normal pituitary in the intrasellar region,and isointensity on T1 and T2 sequence.

Overexpression of EIF5A2 is associated with poor survival and aggressive tumor biology in gallbladder cancer.

Gallbladder cancer (GBC) is a malignant tumor of the biliary tract. The main problem affecting the treatment of gallbladder cancer is late diagnosis and poor prognosis. EIF5A2 is one of two isoforms of the EIF5A family and is reported to be a new oncogenic protein in many human cancers. In this study, our results showed for the first time that EIF5A2 was overexpressed in GBC samples compared with non-tumor tissue. Overexpression of EIF5A2 was associated with lymph node metastasis, tumor differentiation, UICC (Union for International Cancer Control) staging, histological type, metastasis, and tumor size. Overexpression of EIF5A2 in gallbladder carcinoma tissues is also associated with poor prognosis in patients. The interference of EIF5A2 significantly inhibited the proliferation, cell cycle, migration and colony formation of GBC-SD cells in vitro. Our results suggest that EIF5A2 is a target oncogene and may be an important prognostic biomarker in the pathogenesis of gallbladder cancer.

Value of Texture Analysis of Intravoxel Incoherent Motion Parameters in Differential Diagnosis of Pancreatic Neuroendocrine Tumor and Pancreatic Adenocarcinoma.

Objective To evaluate the value of texture features derived from intravoxel incoherent motion (IVIM) parameters for differentiating pancreatic neuroendocrine tumor (pNET) from pancreatic adenocarcinoma (PAC).Methods Eighteen patients with pNET and 32 patients with PAC were retrospectively enrolled in this study. All patients underwent diffusion-weighted imaging with 10 b values used (from 0 to 800 s/mm 2). Based on IVIM model, perfusion-related parameters including perfusion fraction (f), fast component of diffusion (Dfast) and true diffusion parameter slow component of diffusion (Dslow) were calculated on a voxel-by-voxel basis and reorganized into gray-encoded parametric maps. The mean value of each IVIM parameter and texture features [Angular Second Moment (ASM), Inverse Difference Moment (IDM), Correlation, Contrast and Entropy] values of IVIM parameters were measured. Independent sample t-test or Mann-Whitney U test were performed for the between-group comparison of quantitative data. Regression model was established by using binary logistic regression analysis, and receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic efficiency.Results The mean f value of the pNET group were significantly higher than that of the PAC group (27.0% vs. 19.0%, P = 0.001), while the mean values of Dfast and Dslow showed no significant differences between the two groups. All texture features (ASM, IDM, Correlation, Contrast and Entropy) of each IVIM parameter showed significant differences between the pNET and PAC groups (P=0.000-0.043). Binary logistic regression analysis showed that texture ASM of Dfast and texture Correlation of Dslow were considered as the specific imaging variables for the differential diagnosis of pNET and PAC. ROC analysis revealed that multiple texture features presented better diagnostic performance than IVIM parameters (AUC 0.849-0.899 vs. 0.526-0.776), and texture ASM of Dfast combined with Correlation of Dslow in the model of logistic regression had largest area under ROC curve for distinguishing pNET from PAC (AUC 0.934, cutoff 0.378, sensitivity 0.889, specificity 0.854).Conclusions Texture analysis of IVIM parameters could be an effective and noninvasive tool to differentiate pNET from PAC.

Differential Diagnostic Value of Texture Feature Analysis of Magnetic Resonance T2 Weighted Imaging between Glioblastoma and Primary Central Neural System Lymphoma.

Objective To investigate the difference in tumor conventional imaging findings and texture features on T2 weighted images between glioblastoma and primary central neural system (CNS) lymphoma.Methods The pre-operative MRI data of 81 patients with glioblastoma and 28 patients with primary CNS lymphoma admitted to the Chinese PLA General Hospital and Hainan Hospital of Chinese PLA General Hospital were retrospectively collected. All patients underwent plain MR imaging and enhanced T1 weighted imaging to visualize imaging features of lesions. Texture analysis of T2 weighted imaging (T2WI) was performed by use of GLCM texture plugin of ImageJ software, and the texture parameters including Angular Second Moment (ASM), Contrast, Correlation, Inverse Difference Moment (IDM), and Entropy were measured. Independent sample t-test and Mann-Whitney U test were performed for the between-group comparisons, regression model was established by Binary Logistic regression analysis, and receiver operating characteristic (ROC) curve was plotted to compare the diagnostic efficacy.Results The conventional imaging features including cystic and necrosis changes (P=0.000), 'Rosette' changes (P=0.000) and 'incision sign' (P=0.000), except 'flame-like edema' (P=0.635), presented significantly statistical difference between glioblastoma and primary CNS lymphoma. The texture features, ASM, Contrast, Correlation, IDM and Entropy, showed significant differences between glioblastoma and primary CNS lympoma (P=0.006, 0.000, 0.002, 0.000, and 0.015 respectively). The area under the ROC curve was 0.671, 0.752, 0.695, 0.720 and 0.646 respectively, and the area under the ROC curve was 0.917 for the combined texture variables (Contrast, cystic and necrosis, 'Rosette' changes, and 'incision sign') in the model of Logistic regression. Binary Logistic regression analysis demonstrated that cystic and necrosis changes, 'Rosette' changes and 'incision sign' and texture Contrast could be considered as the specific texture variables for the differential diagnosis of glioblastoma and primary CNS lymphoma.Conclusions The texture features of T2WI and conventional imaging findings may be used to distinguish glioblastoma from primary CNS lymphoma.

Value of Magnetic Resonance Imaging Texture Analysis in the Differential Diagnosis of Benign and Malignant Breast Tumors.

Objective To investigate the difference in texture features on diffusion weighted imaging (DWI) images between breast benign and malignant tumors.Methods Patients including 56 with mass-like breast cancer, 16 with breast fibroadenoma, and 4 with intraductal papilloma of breast treated in the Hainan Hospital of Chinese PLA General Hospital were retrospectively enrolled in this study, and allocated to the benign group (20 patients) and the malignant group (56 patients) according to the post-surgically pathological results. Texture analysis was performed on axial DWI images, and five characteristic parameters including Angular Second Moment (ASM), Contrast, Correlation, Inverse Difference Moment (IDM), and Entropy were calculated. Independent sample t-test and Mann-Whitney U test were performed for intergroup comparison. Regression model was established by using Binary Logistic regression analysis, and receiver operating characteristic curve (ROC) analysis was carried out to evaluate the diagnostic efficiency.Results The texture features ASM, Contrast, Correlation and Entropy showed significant differences between the benign and malignant breast tumor groups (PASM=0.014, Pcontrast=0.019, Pcorrelation=0.010, Pentropy=0.007). The area under the ROC curve was 0.685, 0.681, 0.754, and 0.683 respectively for the positive texture variables mentioned above, and that for the combined variables (ASM, Contrast, and Entropy) was 0.802 in the model of Logistic regression. Binary Logistic regression analysis demonstrated that ASM, Contrast and Entropy were considered as the specific imaging variables for the differential diagnosis of breast benign and malignant tumors.Conclusions The texture analysis of DWI may be a simple and effective tool in the differential diagnosis between breast benign and malignant tumors.

[Epidermoid Cyst in the Fourth Ventricle:Report of One Case].

We reported a case with epidermoid cyst (EC) in the fourth ventricle. The insinuation and scalloping growth pattern were the distinctive imaging characteristics of EC in the fourth ventricle on magnetic resonance imaging. The accurate identification of the punctate enhancement of vessels around the lesion may be helpful for its differential diagnosis from hemangioblastoma.

[Value of Magnetic Resonance Imaging Texture Feature Analysis in the Differential Diagnosis between Pancreatic Serous Cystadenoma and Mucinous Cystadenoma].

Objective To investigate the difference of tumor textures presented on T1-weighted image(T1WI) and T2-weighted image(T2WI) between pancreatic serous cystadenoma and mucinous cystadenoma. Methods Subjects including 24 patients with serous cystadenoma and 20 patients with mucinous cystadenoma who were treated in Hainan Branch of Chinese PLA General Hospital were enrolled in this study.The texture analysis was performed on T1WI and T2WI,and the texture parameters included angular second moment,contrast,correlation,inverse difference moment(IDM),and entropy.Independent sample t-test and Mann-Whitney U test were performed for the between-group comparison,the regression model was established by Binary Logistic regression analysis,and receiver operating characteristics curve(ROC) was drawn.Results The contrast and IDM of T1WI and the contrast,correlation,and IDM of T2WI showed significant differences between serous cystadenoma and mucinous cystadenoma(T1WI:Pangular second momont=0.714,Pcontrast=0.003,Pcorrelation=0.069,PIDM=0.004,Pentropy=0.818;T2WI:Pangular second momont=0.122,Pcontrast=0.000,Pcorrelation=0.000,PIDM=0.024,and Pentropy=0.081).The area under the ROC curve was 0.736-0.846 for the positive texture variables,and the area under the ROC curve was 0.879 for the variables in the model of Logistic regression.Binary Logistic regression analysis demonstrated that the contrast was a specific imaging variable for the differential diagnosis between serous cystadenoma and mucinous cystadenoma.Conclusion The texture analysis of T1WI and T2WI may be a simple and effective tool in the differential diagnosis between pancreatic serous cystadenoma and mucinous cystadenoma.

[Magnetic Resonance Imaging Features of the Supratentorial Extra-axial Hemangioblastoma:A Case Report].

We reported a case of supratentorial extra-axial hemangioblastoma diagnosed by magnetic resonance imaging (MRI). As a simple solid mass,this tumor has its unique MRI features due to its tissue structures.

Depside α-glucosidase inhibitors from a culture of the mushroom Stereum hirsutum.

Nine new isoprenylated depsides, sterenins E-M (1-9), as well as five known compounds (10-14), were isolated from the solid culture of Stereum hirsutum. The structures of the new compounds were elucidated by spectroscopic methods. Their inhibitory activities against yeast α-glucosidase were evaluated in vitro. Compounds 1-4 and 7-14 showed inhibitory activities with IC50 values of 7.62, 3.06, 6.03, 22.70, 36.64, 13.09, 27.52, 25.10, 12.32, 3.31, 23.82, and 14.17 µM, respectively. Compounds 5 and 6 showed no inhibitory activities with IC50 values higher than 50 µM. Therefore, the culture of S. hirsutum and its secondary metabolites could have a potential usage for the development of hypoglycemic drugs.

Role of pepsin and pepsinogen: linking laryngopharyngeal reflux with otitis media with effusion in children.

OBJECTIVES/HYPOTHESIS: To analyze the relationship between laryngopharyngeal reflux (LPR) represented by pepsin and pepsinogen, and pathogenesis of otitis media with effusion (OME). STUDY DESIGN: Prospective case-control study. METHODS: Children with OME who required adenoidectomy and tympanostomy/tympanostomy tubes placement were enrolled in OME group, whereas children with adenoid hypertrophy (AH) who required adenoidectomy and individuals who required cochlear implantation (CI) were enrolled in AH and CI groups, respectively. Pepsinogen mRNA and protein levels were assessed by real-time fluorescence-based quantitative polymerase chain reaction and immunohistochemistry in adenoid specimens from the OME and AH groups. Pepsin and pepsinogen concentrations were evaluated by enzyme-linked immunosorbent assay in middle ear fluid and plasma from the OME and CI groups. RESULTS: The levels of pepsinogen protein expressed in cytoplasm of epithelial cells and clearance under epithelial cells in adenoid specimens from the OME group were significantly higher than those in the AH group. Furthermore, the concentrations of pepsin and pepsinogen in the OME group were 51.93±11.58 ng/mL and 728±342.6 ng/mL, respectively, which were significantly higher than those in the CI group (P<.001). In addition, the concentrations of pepsin in dry ears were significantly lower than those in serous and mucus ears in the OME group (F=22.77, P<.001).Finally, the concentration of pepsinogen in middle ear effusion was positively correlated with the expression intensity of pepsinogen protein in cytoplasm of epithelial cells (r=0.73, P<.05) in the OME group. CONCLUSIONS: Pepsin and pepsinogen in middle ear effusion are probably caused by LPR and may be involved in the pathogenesis of OME. LEVEL OF EVIDENCE: 3b.

Effect of nonpeptide NK1 receptor antagonist L-703,606 on the edema formation in rats at early stage after deep partial-thickness skin scalding.

OBJECTIVE: To investigate the effect and the relevant potential mechanism of nonpeptide neurokinin 1 (NK1) receptor antagonist L-703,606 in the edema formation after burn injury. METHOD: L-703,606 treatment was performed in Sprague-Dawley (SD) rats at early stage after deep partial-thickness skin scalding. One hundred and fifty two adult male SD rats were used in the study and randomly divided into sham scald (SS, n=8), scald control (SC, n=48), and L-703,606 treatment (LT, n=48) groups. The rats in SC and LT groups were subjected to 20% total body surface area (TBSA) deep partial-thickness skin scalding. Modified Evans blue extravasation, tracing electron microscopy by lanthanum nitrate and mean water content assay were employed to observe and detect the changes of vascular permeability, ultrastructure and edema formation in adjacent tissue to the wounds and in the jejuna of rats at early stage (72 h) after scald. RESULTS: The pathological increase of vascular permeability in the periwound tissue and jejunum of rats in LT group were significantly lower than that in SC group (P<0.01), and recuperated earlier. Meanwhile, the changes of water contents of corresponding tissues in LT group were lighter than those in SC group (P<0.01). The ultrastructural changes of the microvessels in the peri-wound tissue of LT group showed that the junctions between microvascular endothelium cells were more narrow than those of SC group, moreover, and the number of opening and the engorgement and cavitation of the vascular endothelium cells decreased, the areosis and edema in perivascular tissue lightened, and the precipitation of the high eletron density lanthanum tracing agent in the interspace of the tissue decreased significantly in LT group. CONCLUSIONS: It is concluded that nonpeptide NK1-receptor antagonist L-703,606 could lighten the vascular permeability and edema formation in the periwound tissue and jejunum, and accelerate the normalization process of pathological changes in the tissues of rats after scald.

[Infrared spectral analysis for calcined borax].

To valuate the quality of calcined borax which is sold in the market, 18 samples of calcined borax were studied using the Fourier transform infrared, and samples with different water content were selected and analyzed. Then, the results of analysis were used to evaluate the quality of calcined borax. Results show that the infrared spectra of calcined borax include OH vibration, BO3(-3) vibration and BO4(5-) vibration absorption bands. The position and width of OH vibration absorption band depend on the level of water content, and the more the water content, the wider the absorption band. The number of BO3(3-) vibration and BO4(5-) vibration bands also depend on the level of water content, and the more the water content, and the stronger the hydrogen bond and the lower the symmetry of B atoms, the more the number of infrared absorption peaks. It was concluded that because the quality of calcined borax has direct correlation with water content, the infrared spectroscopy is an express and objective approach to quality analysis and evaluation of calcined borax.

[Similarity analysis of FTIR of Lapis Micae Aureum].

Similarity analysis of FTIR of Lapis Micae Aureum was carried out to find out the relationship between similarity of infrared spectra and the quality of medicine samples, and try to provide a new method for its quality assessment. FTIR was used to analyze Lapis Micae Aureum samples. Then 6 samples with good quality were picked up to establish a reference infrared spectra by their infrared spectra. Correlation coefficient method and the included angle cosine method were used to calculate the similarity between sample's infrared spectra and the reference infrared spectra. For all those 19 samples with characters in accordance with Ch. P, the similarity is above 0.94, and for the others the similarity is less than 0.94. Although some samples' source meets the requirement of Ch. P, but with poor quality by traditional experience, the similarity is lower than those of good quality. It was concluded that the similarity of infrared spectra of Lapis Micae Aureum is closely related to the quality of medicines. As a result, similarity analysis of FTIR of Lapis Micae Aureum is reliable, and can be used for its quality assessment.