Seroprevalence of Pteropine orthoreovirus in humans remain similar after nearly two decades (2001-2002 vs. 2017) in Tioman Island, Malaysia.

Pteropine orthoreovirus (PRV) is an emerging zoonotic respiratory virus that can be transmitted from bats to humans. In Malaysia, aside from PRV2P (Pulau virus) being isolated from Pteropus hypomelanus sampled in Tioman Island, PRV3M (Melaka virus), PRV4K (Kampar virus), and PRV7S (Sikamat virus) were all isolated from samples of patients who reported having a disease spectrum from acute respiratory distress to influenza-like illness and sometimes even with enteric symptoms such as diarrhea and abdominal pain. Screening of sera collected from human volunteers on Tioman Island in 2001-2002 demonstrated that 12.8% (14/109) were positive for PRV2P and PRV3M. Taking all these together, we aim to investigate the serological prevalence of PRV (including PRV4K and PRV7S) among Tioman Island inhabitants again with the assumption that the seroprevalence rate will remain nearly similar to the above reported if human exposure to bats is still happening in the island. Using sera collected from human volunteers on the same island in 2017, we demonstrated seroprevalence of 17.8% (28/157) against PRV2P and PRV3M, respectively. Seropositivity of 11.4% among Tioman Island inhabitants against PRV4K and PRV7S, respectively, was described in this study. In addition, the seroprevalence of 89.5% (17/19), 73.6% (14/19), 63.0% (12/19), and 73.6% (14/19) against PRV2P, PRV3M, PRV4K, and PRV7S, respectively, were observed among pteropid bats in the island. We revealed that the seroprevalence of PRV among island inhabitants remains nearly similar after nearly two decades, suggesting that potential spill-over events in bat-human interface areas in the Tioman Island. We are unclear whether such spillover was directly from bats to humans, as suspected for the PRV3M human cases, or from an intermediate host(s) yet to be identified. There is a high possibility of the viruses circulating among the bats as demonstrated by high seroprevalence against PRV in the bats.

Solid-state fermentation produces greater stearidonic acid levels in genetically engineered Mucor circinelloides.

Omega-3 (ω-3) polyunsaturated fatty acids (PUFAs) are important dietary components due to their health benefits and preventative role in cardiovascular disease. Fish-based and plant seed oils are rich in stearidonic acid (SDA; 18:4, n-3), which are readily metabolized into ω-3 PUFAs such as eicosapentaenoic acid. However, these natural sources of SDA are generally low yielding and are unlikely to meet global demands, so new sustainable microbial fermentative sources of SDA need to be identified. Expression of delta15-desaturase in the oleaginous filamentous fungus Mucor circinelloides (McD15D) has been used to construct a recombinant SDA-producing McD15D strain that produces 5·0% SDA levels using submerged fermentation conditions. Switching to solid-state fermentation conditions in the same medium with submerged fermentation resulted in this engineered strain producing significantly higher amounts of SDA. A Box-Behnken design of response surface methodology approach has been used to identify optimal glucose and ammonium tartrate concentrations and temperature levels to maximize SDA production. The use of these optimal solid-state fermentation conditions resulted in the spores and mycelium of the recombinant McD15D producing 19·5% (0·64 mg g-1 ) and 12·2% (1·52 mg g-1 ) SDA content, respectively, which represents an overall increase in SDA yield of 188·0% compared with SDA yields produced using submerged fermentation conditions.

[Clinicopathological features of hybrid oncocytic/chromophobe renal cell tumor].

Objective: To investigate the clinicopathological features and immunohistochemical phenotypes of hybrid oncocytic/chromophobe tumor (HOCT) of the kidney and its associations with renal oncocytoma (RO) and eosinophilic chromophobe renal cell carcinoma (eChRCC). Methods: A total of 8 HOCT cases were collected from 2008 to 2019 at the Affiliated Hospital of Qingdao University (5 cases) and 971 Hospital of PLA Navy (3 cases), Qingdao, China for morphological studies, immunohistochemical staining and follow-up. The immunohistochemical results of HOCT were compared with those of 27 typical RO and 17 eChRCC. Results: Among the 8 patients, 3 were male and 5 were female. Their ages ranged from 39 to 75 years (median: 56 years). All cases were sporadic. Seven patients were asymptomatic and one suffered from lumbago. During a mean follow-up of 37 months in 7 patients, none of them developed tumor recurrence or metastasis. Seven cases were solitary and one was multiple. The tumor size ranged from 1.4 to 5.7 cm (mean, 3.6 cm). The cut surface of the tumors was dark red or yellowish. Histologically, the tumors were well-defined. Six cases were directly adjacent to the surrounding renal tissue, 2 cases had pseudocapsule, 3 cases showed entrapped renal tubules at the edge of tumor tissue, and one circumscribed with focal infiltrating borders. There were two types of histological morphology: one type (4 cases) was composed of mixed areas of otherwise typical RO and areas resembling chromophobe renal cell carcinoma; another type (4 cases) showed the morphological characteristics of both RO and eChRCC. Three second-type tumors showed nest-like, trabecular, and solid growth patterns with conspicuous edematous stroma. The cell border was conspicuous and the cytoplasm showed an eosinophilic appearance. The nuclei were small and round with clear perinuclear halo. One tumor showed a multi-nodular and solid growth pattern, and the cytoplasm was eosinophilic, hypochromatic or transparent. The nuclei were small and round, and some of them had obvious perinuclear halo. Immunohistochemically, the tumor cells in all 8 cases were positive for Ksp-cad but negative for vimentin. CD117 was diffusely positive in 6/8 cases. CK7 staining showed patchy positivity in 6/8 cases. S-100A1, cyclin D1 and claudin7 showed variable positivity in 4/8, 6/8 and 5/8 cases, respectively, but the range and intensity were narrower and weaker than those in RO and eChRCC. Conclusions: HOCT is a low-grade eosinophilic renal tumor with morphological characteristics resembling RO and eChRCC. The combined application of immunohistochemical stains of CK7, CD117, Ksp-cad, cyclin D1, claudin7 and S-100A1 may play an auxiliary role in the differentiation of the three tumors. HOCT has a good prognosis after surgical resection and can be regarded as a tumor with uncertain malignant potential.

[Clinicopathological features and gene phenotypes of benign metastasizing leiomyoma].

Objective: To study the clinicopathological features, immunophenotypes and MED12 gene status in benign metastasizing leiomyoma (BML). Methods: Nine cases of BML diagnosed at the Affiliated Hospital of Qingdao University from 2012 to 2018 were collected, and the radiologic and histologic features were analyzed. The protein expression of leiomyosarcoma-related driver genes, including RB1, PTEN,ATRX,p16,p53, as well as ER,PR,CD34,FH, and Ki-67 were detected using immunohistochemistry, and the mutation status of MED12 gene exon 2 was detected by Sanger sequencing. Results: All the nine patients with BML were female, and the age range was 48 to 64 years (median 55 years). All patients had history of uterine fibroids. The morphologic features of BML were similar to a benign uterine leiomyoma and did not exhibit malignant characteristics. All cases were positive for ER and PR, and negative for CD34. In addition, RB1, PTEN, ATRX, and FH were positive in all cases (wild type), while p16 showed a focally positive pattern. P53 positive index was less than 5% (wild type), and Ki-67 positive index was less than 1%. Sanger sequencing was done in six BML samples; one sample harbored a nonsense mutation c. 142_144delinsTAA (p.Glu48Ter), and another exhibited a synonymy mutation (c.192C>T, p.Phe64=)and one missense mutation c.196C>T (p.Pro66Ser). Conclusions: The present study suggests that BML is a unique leiomyoma entity that is pathologically and genetically different from leiomyosarcomas and conventional uterine leiomyomas. Evaluating the genetic phenotype of BML, especially the expression of leiomyosarcoma-related driver genes protein and MED12 gene status, may be helpful in understanding the pathogenesis of BML and in its differentiation from leiomyosarcoma.

[Establishment and clinical testing of pancreatic cancer Faster R-CNN AI system based on fast regional convolutional neural network].

Objective: To investigate the effectiveness of an enhanced CT automatic recognition system based on Faster R-CNN for pancreatic cancer and its clinical value. Methods: In this study, 4 024 enhanced CT imaging sequences of 315 patients with pancreatic cancer from January 2013 to May 2016 at the Affiliated Hospital of Qingdao University were collected retrospectively, and 2 614 imaging sequences were input into the faster R-CNN system as training dataset to create an automatic image recognition model, which was then validated by reading 1 410 enhanced CT images of 135 cases of pancreatic cancer.In order to identify its effectiveness, 3 750 CT images of 150 patients with pancreatic lesions were read and a followed-up was carried out.The accuracy and recall rate in detecting nodules were recorded and regression curves were generated.In addition, the accuracy, sensitivity and specificity of Faster R-CNN diagnosis were analyzed, the ROC curves were generated and the area under the curves were calculated. Results: Based on the enhanced CT images of 135 cases, the area under the ROC curve was 0.927 calculated by Faster R-CNN. The accuracy, specificity and sensitivity were 0.902, 0.913 and 0.801 respectively.After the data of 150 patients with pancreatic cancer were verified, 893 CT images showed positive and 2 857 negative.Ninety-eight patients with pancreatic cancer were diagnosed by Faster R-CNN.After the follow-up, it was found that 53 cases were post-operatively proved to be pancreatic ductal carcinoma, 21 cases of pancreatic cystadenocarcinoma, 12 cases of pancreatic cystadenoma, 5 cases of pancreatic cyst, and 7 cases were untreated.During 5 to 17 months after operation, 6 patients died of abdominal tumor infiltration, liver and lung metastasis.Of the 52 patients who were diagnosed negative by Faster R-CNN, 9 were post-operatively proved to be pancreatic ductal carcinoma. Conclusion: Faster R-CNN system has clinical value in helping imaging physicians to diagnose pancreatic cancer.

TTK contributes to tumor growth and metastasis of clear cell renal cell carcinoma by inducing cell proliferation and invasion.

The aim of this study was to determine the expression levels of TTK in clear cell renal cell carcinoma (ccRCC) tissues and its possible link with the clinical pathologic characteristics and the prognosis of patients suffering this disease, and to further explore the potential role of TTK in the progression of ccRCC. Immunohistochemical (IHC) assays were performed to detect the expression levels of TTK in 112 samples of ccRCC tissues and corresponding non-tumor tissues. According to the results of IHC assays, patients were divided into TTK high expression and low expression group. Clinical analysis related to the clinical features (age, gender, T stage), and the potential link between TTK expression levels and clinical features were analyzed. In addition, the effects of TTK on the proliferation and invasion of ccRCC cells were detected through colony formation assay and transwell assays, respectively. The possible effects of TTK on tumor growth and metastasis were measured in mice. We found a high expression level of TTK in human ccRCC tissues from patients who received surgical treatment. We also found its expression level was obviously associated with clinical characteristics, such as T stage (p=0.008) and lymphatic metastasis (p=0.023). We further confirmed that knockdown of TTK suppressed cell proliferation and invasion in 2 types of ccRCC cells, HTB-47 and CRL-1932 cells. Furthermore, TTK contributes to tumor growth and metastasis of ccRCC in mice. We found that TTK affected the progression of ccRCC and further mechanically confirmed it could be a novel therapeutic target for ccRCC treatment.

[Clinical application of convolutional neural network in pathological diagnosis of metastatic lymph nodes of gastric cancer].

Objective: To examine the value and clinical application of convolutional neural network in pathological diagnosis of metastatic lymph nodes of gastric cancer. Methods: Totally 124 patients with advanced gastric cancer who underwent radical gastrectomy plus D2 lymphadenectomy at Affiliated Hospital of Qingdao University from July 2016 to December 2018 were selected in the study. According to the chronological order, the first 80 cases were served as learning group. The remaining 44 cases were served as verification group. There were 45 males and 35 females in the study group, with average age of 57.6 years. There were 29 males and 15 females in the validation group, with average age of 9.2 years. The pre-training convolutional neural network architecture Resnet50 was trained and fine-tuned by 21 352 patches with cancer areas and 14 997 patches without cancer areas in the training group. A total of 78 whole-slide image served as a test dataset including positive (n=38) and negative (n=40) lymph nodes. The convolutional neural network computer-aided detection (CNN-CAD) system was used to analyze the ability of convolutional neural network system to screen metastatic lymph nodes at the level of slice by setting threshold, and evaluate the system's classification accuracy by calculating its sensitivity, specificity, positive predictive value, negative predictive value and area under the receiver operating characteristic curve (AUC). Results: The classification accuracy of CNN-CAD system at slice level was 100%.The AUC for the CNN-CAD system was 0.89. The sensitivity was 0.778, specificity was 0.995, overall accuracy was 0.989. Positive and negative predictive values were 0.822 and 0.994, respectively. The CNN-CAD system achieved the same classification results as pathologists. Conclusions: The CNN-CAD system has been constructed to distinguished benign and malignant lymph node slides with high accuracy and specificity. It could achieve the similar classification results as pathologists.

Differential expression and analysis of target regulation of microRNAs in alcohol-dependent rats.

This study analyzed the differential expression of miRNAs related to alcohol dependence in rats undergoing continued consumption and withdrawal. Furthermore miRNAs were sought and evaluated for potential use as biomarkers for diagnosis. This study used Exiqon miRCURYTM LNA miRNA microarray on alcohol-dependent and normal rats for the expression of microRNAs in perfluorinated compounds, and the stem-loop qPCR method to validate its expression in brain tissue. We forecast the relevant target genes of differentially expressed miRNA and drew the regulatory network. Comparison of the differential expression between brain tissue and plasma was carried out and the correlation and analyzed.65 miRNA with differential expression with threshold of 1.5 were screened out; among them, most miRNA with differential expression in the dependent group had relatively high expression values. The target genes were found with great confidence: PIK3CA, MAPK, NTF, BDNF, NGFR, IGF-1, and the pair consisting of miRNA- mRNA. Among the three groups, the expression levels of miR-101b (F=8.12, P less than0.05) showed significant difference; no significant difference was found in the expression levels of miRNA in plasma among the groups (F=1.23, P>0.05). No consistency was shown in the changing trend of miRNA in PFC and plasma (r= -.004, p>0.05). The genetic regulatory network of neurotrophic factors, its receptors and the protein kinases that influence metabolism may mediate the incidence of alcohol-dependence. There is a lack of conformity between the expression of miR-101b in the prefrontal cortex and the plasma.

Biodegradation of diethyl terephthalate and polyethylene terephthalate by a novel identified degrader Delftia sp. WL-3 and its proposed metabolic pathway.

Polyethylene terephthalate (PET), a synthetic polyester material made of diethyl terephthalate (DET) monomers, is widely used in plastic products of daily life and caused serious pollution to the global environment. Microbial metabolism is the major degradation pathway responsible for DET degradation in natural soil; however, the microbial DET degradation mechanism remains unclear. In this study, the newly isolated strain WL-3, identified as belonging to the genus Delftia, was found to be able to degrade 94% of 5 g l-1 of DET and utilize it as the sole carbon source for growth within 7 days. Furthermore, strain WL-3 was capable of stable DET degradation under a wide range of pH values (6·0-9·0) and temperatures (20-42°C) with the optimal pH and temperature of 7·0 and 30°C respectively. Furthermore, the biochemical pathway of DET degradation by strain WL-3 was proposed based on the identified degradation intermediates. DET is first transformed into terephthalic acid (TPA) by the hydrolysis of two ester bonds, which is subsequently converted to protocatechuic acid (PCA) and further mineralized. SEM observations revealed obvious cracks on the surface of PET film after inoculation of 2 months with strain WL-3, indicating the strain's potential for the bioremediation of PET-contaminated environments. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates that Delftia sp. WL-3 can mineralize completely diethyl terephthalate by biochemical processes. The study reveals the metabolic mechanism of diethyl terephthalate biodegradation. Furthermore, the cracks on the surface of Polyethylene terephthalate film that form upon inoculation with strain WL-3 were observed using SEM. These results highlight the potential of the strain WL-3 in the bioremediation of environments contaminated with Polyethylene terephthalate or diethyl terephthalate.

[An interpretation of European guidelines for the diagnosis and treatment of nonalcoholic fatty liver disease].

Nonalcoholic fatty liver disease (NAFLD) is a pathological manifestation of obesity and metabolic syndrome involving the liver. It can cause disability and mortality related to liver disease, and it is closely associated with the high incidence rates of type 2 diabetes and cardiovascular diseases. At present, NAFLD has become the most common chronic liver disease worldwide, and the development and implementation of the clinical guideline for the diagnosis and treatment of NAFLD help to improve patients' prognosis. This article provides an interpretation of the latest Europeans guidelines through a comparative analysis of the previous and latest Europeans guidelines.

Clinical significance of serum miR-196a in cervical intraepithelial neoplasia and cervical cancer.

Previous studies have reported that miR-196a is upregulated in cervical cancer tissues and cell lines. However, whether serum miR-196a is increased in patients with cervical cancer or cervical intraepithelial neoplasia (CIN), and its potential clinical value remained unknown. In total, 105 cervical cancer patients, 86 CIN patients, and 50 healthy volunteers were recruited. Quantitative reverse transcription-polymerase chain reaction was performed to compare the serum levels miR-196a in all participants. The associations between serum miR-196a and CIN grade/clinicopathological parameters of cervical cancer were also examined. A survival analysis was performed using the Kaplan-Meier method. Univariate and multivariate analyses were conducted to explore the independent risk factors for cervical cancer. Our results revealed that serum miR-196a levels were higher in patients with cervical cancer (P < 0.01) and CIN (P < 0.05) compared to those in healthy controls. Serum miR-196a was associated with CIN grade and various cervical cancer parameters including tumor size (P = 0.031), lymph node metastasis (P = 0.018), FIGO stage (P = 0.004), and grade (P = 0.011). Cervical cancer patients with higher serum miR-196a levels had a poorer overall survival rate (P = 0.004). Multivariate analysis revealed that high serum miR-196a was an independent predictor for poor survival of cervical cancer (HR = 3.510; 95%CI = 1.961-6.874; P = 0.025). In conclusion, our findings suggest that serum miR-196a overexpression is associated with CIN grade and cervical cancer progression. Therefore, serum miR-196a may be a reliable biomarker for early detection and prognosis of cervical cancer.

A 3-D elasticity theory based model for acoustic radiation from multilayered anisotropic plates.

A theoretical model built upon three-dimensional elasticity theory is developed to investigate the acoustic radiation from multilayered anisotropic plates subjected to a harmonic point force excitation. Fourier transform technique and stationary phase method are combined to predict the far-field radiated sound pressure of one-side water immersed plate. Compared to equivalent single-layer plate models, the present model based on elasticity theory can differentiate radiated sound pressure between dry-side and wet-side excited cases, as well as discrepancies induced by different layer sequences for multilayered anisotropic plates. These results highlight the superiority of the present theoretical model especially for handling multilayered anisotropic structures.

Effects of mean flow on transmission loss of orthogonally rib-stiffened aeroelastic plates.

This paper investigates the sound transmission loss (STL) of aeroelastic plates reinforced by two sets of orthogonal rib-stiffeners in the presence of external mean flow. Built upon the periodicity of the structure, a comprehensive theoretical model is developed by considering the convection effect of mean flow. The rib-stiffeners are modeled by employing the Bernoulli-Euler beam theory and the torsional wave equation. While the solution for the transmission loss of the structure based on plate displacement and acoustic pressures is given in the form of space-harmonic series, the corresponding coefficients are obtained from the solution of a system of linear equations derived from the plate-beam coupling vibration governing equation and Helmholtz equation. The model predictions are validated by comparing with existing theoretical and experimental results in the absence of mean flow. A parametric study is subsequently performed to quantify the effects of mean flow as well as structure geometrical parameters upon the transmission loss. It is demonstrated that the transmission loss of periodically rib-stiffened structure is increased significantly with increasing Mach number of mean flow over a wide frequency range. The STL value for the case of sound wave incident downstream is pronouncedly larger than that associated with sound wave incident upstream.

Gene expression profiling of hybridoma cells after bursal-derived bioactive factor BP5 treatment.

Bursa of Fabricius is the acknowledged vital humoral immune system for B cell differentiation and antibody production. To study the molecular mechanism underlying the effect of bursal-derived BP5, we used gene microarray to analyze the genomic expression profiling of BP5-treated hybridoma cells. BP5 exhibited an immunomodulatory effect on antibody production in hybridoma cells and induced alterations in the gene expression profiles related to the immune-related biological processes, such as T cell activation and proliferation, B cell activation, B cell-mediated immunity, and cytokines cytokine production involved in immune response. In addition, 26 biological pathways associated with immunomodulatory functions were regulated in BP5-treated hybridoma cells, in which p53 signal pathway played an important role in antitumor. Among these regulated genes, 12 differentially expressed genes were verified by qRT-PCR. The activation of p53 activity by BP5 was further confirmed by p53 luciferase reporter assay and p53 expression. Our data revealed that bursal-derived BP5 could regulate various immune-related cellular processes, including antitumor factor p53 signal pathway, perhaps partially accounting for the reported immunomodulatory roles and novel antiproliferation on tumor cells functions of bursal-derived bioactive factor BP5.

In Situ Observation of Liquid Solder Alloys and Solid Substrate Reactions Using High-Voltage Transmission Electron Microscopy.

The complex reaction between liquid solder alloys and solid substrates has been studied ex-situ in a few studies, utilizing creative setups to "freeze" the reactions at different stages during the reflow soldering process. However, full understanding of the dynamics of the process is difficult due to the lack of direct observation at micro- and nano-meter resolutions. In this study, high voltage transmission electron microscopy (HV-TEM) is employed to observe the morphological changes that occur in Cu6Sn5 between a Sn-3.0 wt%Ag-0.5 wt%Cu (SAC305) solder alloy and a Cu substrate in situ at temperatures above the solidus of the alloy. This enables the continuous surveillance of rapid grain boundary movements of Cu6Sn5 during soldering and increases the fundamental understanding of reaction mechanisms in solder solid/liquid interfaces.

Fat-1 transgenic cattle as a model to study the function of ω-3 fatty acids.

ω-3 polyunsaturated fatty acids have been shown to play an important role in health. Enriched with ω-3 polyunsaturated fatty acids modulate expression of a number of genes with such broad functions as cell proliferation, growth and apoptosis and cell signaling and transduction, these effects, seem to regulate coronary artery disease, hypertension, atherosclerosis, psychiatric disorders and various cancer. In this context, fat-1 transgenic cattle was designed to convert ω-6 to ω-3 fatty acids could form an ideal model to study the effect of ω-3 fatty acids on the above functions. This study focuses on the total genomic difference of gene expression between fat-1 transgenic cattle and wild-type using cDNA microarrays, several genes were found to be overexpressed or suppressed in transgenic cattle relative to wild-type, these discrepancy genes related with lipid metabolism, immunity, inflammation nervous development and fertility.

Transmission loss of orthogonally rib-stiffened double-panel structures with cavity absorption.

The transmission loss of sound through infinite orthogonally rib-stiffened double-panel structures having cavity-filling fibrous sound absorptive materials is theoretically investigated. The propagation of sound across the fibrous material is characterized using an equivalent fluid model, and the motions of the rib-stiffeners are described by including all possible vibrations, i.e., flexural displacements, bending, and torsional rotations. The effects of fluid-structure coupling are account for by enforcing velocity continuity conditions at fluid-panel interfaces. By taking full advantage of the periodic nature of the double-panel, the space-harmonic approach and virtual work principle are applied to solve the sets of resultant governing equations, which are eventually truncated as a finite system of simultaneous algebraic equations and numerically solved insofar as the solution converges. To validate the proposed model, a comparison between the present model predictions and existing numerical and experimental results for a simplified version of the double-panel structure is carried out, with overall agreement achieved. The model is subsequently employed to explore the influence of the fluid-structure coupling between fluid in the cavity and the two panels on sound transmission across the orthogonally rib-stiffened double-panel structure. Obtained results demonstrate that this fluid-structure coupling affects significantly sound transmission loss (STL) at low frequencies and cannot be ignored when the rib-stiffeners are sparsely distributed. As a highlight of this research, an integrated optimal algorithm toward lightweight, high-stiffness and superior sound insulation capability is proposed, based on which a preliminary optimal design of the double-panel structure is performed.