The purpose of this study is to systematically determine an optimal percentile cutoff in histogram analysis for calculating the mean parameters obtained from a nonGaussian continuous-time random-walk (CTRW) diffusion model for differentiating individual glioma grades. This retrospective study included 90 patients with histopathologically proven gliomas (42 grade II, 19 grade III, and 29 grade IV). We performed diffusion-weighted imaging using 17 b-values (0-4000 s/mm²) at 3T, and analyzed the images with the CTRW model to produce an anomalous diffusion coefficient (D m ) along with temporal (α) and spatial (β) diffusion heterogeneity parameters. Given the tumor ROIs, we created a histogram of each parameter; computed the P-values (using a Student’s t-test) for the statistical differences in the mean D m , α, or β for differentiating grade II vs. grade III gliomas and grade III vs. grade IV gliomas at different percentiles (1% to 100%); and selected the highest percentile with P < 0.05 as the optimal percentile. We used the mean parameter values calculated from the optimal percentile cut-offs to do a receiver operating characteristic (ROC) analysis based on individual parameters or their combinations.We compared the results with those obtained by averaging data over the entire region of interest (i.e., 100th percentile). We found the optimal percentiles for D m , α, and β to be 68%, 75%, and 100% for differentiating grade II vs. III and 58%, 19%, and 100% for differentiating grade III vs. IV gliomas, respectively. The optimal percentile cut-offs outperformed the entire-ROI-based analysis in sensitivity (0.761 vs. 0.690), specificity (0.578 vs. 0.526), accuracy (0.704 vs. 0.639), and AUC (0.671 vs.0.599) for grade II vs. III differentiations and in sensitivity (0.789 vs. 0.578) and AUC (0.637 vs. 0.620) for grade III vs. IV differentiations, respectively. Percentile-based histogram analysis, coupled with the multi-parametric approach enabled by the CTRW diffusion model using high b-values, can improve glioma grading.

Objective: To investigate the role of long non-coding RNA double homeobox A pseudogene 9 (DUXAP9) in head and neck squamous cell carcinoma (HNSCC) and to evaluate the expression level, molecular function and mechanism of DUXAP9 in HNSCC cells. Methods: Differential expression of lncRNAs between normal and tumor tissues in HNSCC tissues were screened using lncRNA microarray, the expression level of DUXAP9 in HNSCC tissues and its relationship with prognosis were analyzed in the TCGA database. The expression levels of DUXAP9 in HNSCC tissues and cell lines were detected using qRT-PCR. The function in HNSCC cells after DUXAP9 silencing was evaluated using the CCK-8 assay, wound healing assay, Transwell migration assay and subcutaneous xenograft assay in nude mice. Changes in the transcription and translation of epithelial-mesenchymal transition (EMT)-related proteins in head and neck squamous cell carcinoma cells after DUXAP9 silencing were detected using qRT-PCR and Western blot. Results: lncRNA microarray results showed that, compared to adjacent normal tissues, DUXAP9 was abnormally upregulated in HNSCC tissues. Analysis from TCGA database showed that, compared to HNSCC patients with low DUXAP9 expression, HNSCC patients with high DUXAP9 expression had poorer survival. The relative expression of DUXAP9 in HNSCC tissues and 4 HNSCC cell lines increased compared to paired adjacent normal tissues as detected using qRT-PCR. Silencing DUXAP9 significantly inhibited the proliferation, migration and expression of EMT-related genes in HNSCC cells. The silencing of DUXAP9 significantly inhibited subcutaneous tumorigenesis of the HNSCC cell line CAL27 in nude mice. Conclusion Silencing DUXAP9 significantly inhibited the proliferation of HNSCC cells and subcutaneous xenografts in nude mice. DUXAP9 may mediate the migration of head and neck squamous cell carcinoma cells via the EMT pathway.

Objective To study the influences on the production of major inflammatory cytokines after co-culturing macrophages with human umbilical vein endothelial cells ( HUVECs) that were infected with dengue virus type 2 (DENV-2). Methods Density gradient centrifugation was used to isolate periph-eral blood mononuclear cells ( PBMC) from concentrated human leukocytes. Adherent monocytes in culture flasks were obtained and stimulated with macrophage colony-stimulating factor ( M-CSF) to prepare macro-phages. The purity of CD14+CD11b+ cells was measured by flow cytometry. Changes in the expression of NS1 at mRNA level in HUVECs were detected by real-time PCR following DENV-2 infection. DENV-2-in-fected HUVECs were co-culture with macrophages in Transwell chambers. A control group was set up by pre-treating HUVECs with sphingosine-1-phosphate (S1P) type 1 (S1P1)-specific receptor agonist CYM-5442 for 24 h to remove the drug before infection and then co-culturing the infected cells with macrophages. Real-time PCR was used to detect the expression at mRNA level of IL-6 and IL-8 in HUVECs and IL-6, IL-8, TNF-α and IL-1β in macrophages. A double-antibody sandwich ELISA was used to detect the expression of above cytokines in culture supernatants. Results After HUVECs were infected with DENV-2, expression of NS1 gene at mRNA level gradually increased to the peak at 24 h (2. 66±0. 53, P<0. 05) and then de-creased. The purity of macrophages detected by flow cytometry was (89. 16±2. 07) ％. Expression of IL-6 and IL-8 at mRNA level in DENV-2-infected HUVECs was up-regulated. The peak values reached at 24 h of IL-6 and IL-8 expression were 16. 10±0. 17 and 29. 76±0. 58, while the expression levels at 24 h in the un-infected group were 1. 46±0. 67 and 1. 60±0. 54, respectively. Expression of IL-6, IL-8, TNF-αand IL-1βat mRNA level in DENV-2-infected macrophages was increased significantly. The levels of IL-6, IL-8, TNF-αand IL-1β expression at 24 h were 45. 82±3. 72, 52. 34±1. 69 (12 h), 8. 94±1. 75 and 30. 96±1. 44 in the infected macrophages, and 1. 16±0. 22, 1. 15±0. 21, 1. 11±0. 09 and 1. 47±0. 31 in the uninfected group. Expression of these cytokines was decreased at every time points after co-culturing of DENV-2-infec-ted HUVECs with macrophages, but still significantly higher than that in the uninfected group. In the co-cul-ture group with DENV-2 infection, CYM-5442 pretreatment significantly decreased the expression at mRNA level of IL-6 and IL-8 in HUVECs (P<0. 01) and that of IL-6, IL-8, TNF-αand IL-1βin macrophages (P<0. 01). Conclusions DENV-2 could infect primary HUVECs, and then activate macrophages to promote the secretion of large amounts of IL-6, IL-8, TNF-αand IL-1β. Moreover, the activated macrophages could reduce the production of inflammatory cytokines in HUVECs to a certain extent.

To develop a clinical diagnosis technique for bluetongue virus infection, we established serotype-specific methods to detect serotype 4 of bluetongue virus (BTV-4). Two monoclonal antibodies (mAbs) against VP2 protein of BTV-4, named 4A-1G7 and 4B-1B6, were used as competitive antibodies in the competitive enzyme-linked immunosorbent assays (C-ELISA). We detected 50 negative serum samples from sheep, goats and cattle by C-ELISA. The cut-off values of 4A-1G7 and 4B-1B6 mAbs were 49% and 40%, respectively. The results of the sensitivity, specificity and repeatability by detecting standard positive serum, were consistent with the general standard of Office International Des Epizooties. Furthermore, serum samples of BTV-4, BTV-18 and BTV-20 infection could be screened out through the combined C-ELISAs by 4A-1G7 and 4B-1B6 mAbs. Thus, this technique may diagnose BTV-4, BTV-18 and BTV-20 infections.

To confirm the B cell epitope recognized by monoclonal antibody (MAb) 3G11 of bluetongue virus type 8 (BTV-8) VP2 protein prepared in our laboratory, antigen epitopes recognized by 3G11 were screened and identified by phage display technology. KLLAT sequence was found by sequencing of blue spot after four rounds panning and 283LL284 of common short peptide sequence was obtained after comparison to amino acid sequence of BTV-8 VP2 protein. The peptide sequences KLLAA, KALAT, KLAAT and KLLAT were synthesized and identified by indirect ELISA. KLLAA and KLLAT bound strongly with supernatant and as cites of 3G11 cells and reacted specifically with BTV-8 positive standard sera. Further sequence analysis showed that amino acid sequence 283LL284 was conserved among different serotypes of BTV-8 strains, and283LL284 was the key amino acids of antigen epitopes recognized by 3G11. This study laid the foundation to establish type 8 BTV specific immunological detection methods.

Aortic dissection is a dangerous and critical disease with extremely high mortality and disability rate.In clinical practice,aortic dissection should be highly suspected when patients developed dying-like severe chest and back pain.CT and MRI have been the reliable tools for diagnosing aortic dissection.In recent years,endovascular therapy has become the preferred treatment for Stanford type B aortic dissection,and some patients with Stanford type A dissection who cannot receive open surgery may also be treated with endovascular therapy.In order to improve endovascular treatment,to develop new instruments and to study the pathogenesis of aortic dissection,the preparation of stable and reliable animal models is very necessary.This paper aims to make a brief review about the research status concerning the preparation of animal models of aortic dissection.

Objective: To explore the electrocardiographic characteristics of patients with idiopathic ventricular arrhythmias (VAs) originating from different portions of distal great cardiac veins (DGCV). Methods: The study included 49 patients underwent successful RFCA of premature ventricular complex(PVCs)/ventricular tachycardia(VT) from different portions of the DGCV in our department from July 2009 to March 2016. The surface 12-lead electrocardiogram (ECG) and intraventricular ablation mapping features were analyzed. Patients were divided into four groups according to the mapping and ablation results: DGCV1(10 patients), DGCV2 (13 patients), proximalanterior interventricular vein (PAIV, 17 patients)and extend distal great cardiac vein (EDGCV, 9 patients). We analyzed the similarities and differences between surface 12-lead ECG of patients with PVCs/VT from different portions of DGCV, and compared with random chosen 290 patients with PVCs/VT from ventricular outflow tract and adjacent structure. Results: A positive R wave in inferior leads, a negative QS morphology in lead aVL and aVR were found among all groups. The different characteristics of surface 12-lead ECG of VAs originating from DGCV were as follows: (1)EDGCV patients demonstrated a positive R or r wave on lead Ⅰ(6/9) while a negative rS or qr wave was evidenced in other three groups (39/40). (2)A positive R pattern on lead V₁, V₅-V₆ (11/13) was presented in patients of DGCV2 group; R (without S or s) wave on V₁ (9/10), RS or Rs wave on V₅-V₆ were found in DGCV1 group; RS or rS wave was seen on lead V₁, R(without S)wave in lead V₅-V₆ (25/26) were found in EDGCV and PAIV group and the amplification of R wave in EDGCV was higher than V₁ of PAIV group.(3)Precordial lead transition zone was in front of V₁ for DGCV1 and DGCV2 groups (23/23), within V₁-V₃ for EDGCV group, but on V₂ or within V₂-V₃ for PAIV group.(4)Patients of DGCV1 and DGCV2 demonstrated a longer Pseudo delta wave time(PdW), intrinsicoid deflection time (IDT), significantly larger maximum deflection index (MDI) than those in PAIV and EDGCV groups (P<0.001). (5)The different characteristics of surface 12-lead ECG between VAs originating from DGCV and those from ventricular outflow tract and adjacent structure were as follows: ① The ECG features were similar between PVIA and LCC group, both demonstrated a rs wave on the lead Ⅰ, rS wave on V₁-V₂ and R wave on V₅-V₆; ②The ECG features were similar betweenEDGCV and RCC group, both presented with R or r wave on the lead Ⅰ, the QRS wave of precordial leads was similar as PAIV and LCC groups; ③A R wave on the lead V₁, V₅-V₆ was found in group DGCV2, and ILCC; ④Similar to the group Endo-MAA, patients in DGCV1 group also demonstrated a R wave on the lead V₁ and a Rs wave on V₅-V₆. Conclusion A positive R wave in inferior leads, a negative QS morphology in lead aVL and aVR are seen in all patients, but different electrocardiographic characteristics of PVC/VT originating from the different portions of the DGCV are presented on lead Ⅰ and V₁-V₆.

Objective To establish a scientific and objective indicator system for nursing quality evaluation according to nursing works of neonatal surgical department. Methods To establish the indicator system based on the three- dimension quality structure. Literature retrieval, expert interview and Delphi expert enquiry were used to filter and improve indicators. Analytic hierarchy process was used to determine the index weight and check the consistency. Results The quality indicator system included three levels. There were 3 indicators in level one, 14 indicators in level two and 70 indicators in level three. Experts coefficient was 0.867, mean of importance values of two rounds were 3.83-5.00 and 4.08-5.00 , Kendall coordination coefficients were 0.292(P<0.01) and 0.301(P<0.01). Conclusions The nursing quality evaluation indicator system has high reliability and it can be used for guiding continuous improvement of nursing in neonatal surgical department.

Purpose To evaluate the application value of using a-cyanoacrylate rapid medical glue in preoperative localization of ground-glass nodules under CT guidance.Materials and Methods 48 cases were retrospectively analyzed,in which the pulmonary ground-glass nodules took preoperative localization under CT guidance.The rapid medical glue was injected in pulmonary ground-glass nodules,which was used for preoperative localization.Results After preoperative localization of rapid medical glue in 48 cases,pulmonary ground-glass nodules of all patients were resected successfully by video-assisted thoracoscope surgery (VATS).The complications of pneumothorax did not occur in all cases,with little pulmonary hemorrhagein in 10 cases.Conclusion When the fast medical glue has been used in the CT-guided preoperative localization of ground-glass nodules,there are advantages of high accuracy of localization and surgery.Moreover,this method is simple,safe and effective.

Objective The purpose of this study was to construct a rat model of luteal phase defect ( LPD) with mifepristone ( RU486 ) and investigate the improving effect of Bushen Zhuyun Recipe ( BZR ) on the gonad axis morphology of the LPD rat . Meth ods We established rat models of LPD by giving RU486 at 1 mL/100 g followed by treatment with BZR at 0.4 (medium dose) or 1.2 mL/100g ( high dose ) .Then we observed the changes of morphology of the hypothalamus pituitary ovary uterus . Results Electron microscopy revealed hypothalamic mitochondria vacuolization , degranulation of rough endoplasmic reticulum , dissolved nuclear membrane , and decreased secretory granules , while light microscopy exhibited slightly reduced density of pituitary cells , karyopyknosis of some of the cells ,and cytoplasmic vacuolization in the model rats .Electron microscopy also showed obvious shrinkage of cell nuclei , chromatin condensation , disordered cell arrangement , swelling mitochondrial cavity , and formation of vacuoles in the pituitary anterior lobe, while light microscopy also manifested increased ovarian follicular atresia , reduced granule cell layers and corpora lutea , thinned endometrial layers , decreased uterine glands of the tube wall , reduced glandular cavity , and increased interstitial collagen fibers .After BZR treatment, the number and volume of corpora lutea were increased , the granulosa cell layer and endometrial layer thickened , the number of uterine glands increased , and the gland cavity expanded . Conclusion The axes of the hypothalamus , pituitary, uterus,and o -varian were morphologically changed in the RU 486-induced rat models of LPD, while BZR could improve these changes .