Knockout of the WD40 domain of ATG16L1 enhances foot and mouth disease virus replication.

The WD40 domain is one of the most abundant domains and is among the top interacting domains in eukaryotic genomes. The WD40 domain of ATG16L1 is essential for LC3 recruitment to endolysosomal membranes during non-canonical autophagy, but dispensable for canonical autophagy. Canonical autophagy was utilized by FMDV, while the relationship between FMDV and non-canonical autophagy is still elusive. In the present study, WD40 knockout (KO) PK15 cells were successfully generated via CRISPR/cas9 technology as a tool for studying the effect of non-canonical autophagy on FMDV replication. The results of growth curve analysis, morphological observation and karyotype analysis showed that the WD40 knockout cell line was stable in terms of growth and morphological characteristics. After infection with FMDV, the expression of viral protein, viral titers, and the number of copies of viral RNA in the WD40-KO cells were significantly greater than those in the wild-type PK15 cells. Moreover, RNA‒seq technology was used to sequence WD40-KO cells and wild-type cells infected or uninfected with FMDV. Differentially expressed factors such as Mx1, RSAD2, IFIT1, IRF9, IFITM3, GBP1, CXCL8, CCL5, TNFRSF17 were significantly enriched in the autophagy, NOD-like receptor signaling pathway, RIG-I-like receptor signaling pathway, Toll-like receptor signaling pathway, cytokine-cytokine receptor interaction and TNF signaling pathway, etc. The expression levels of differentially expressed genes were detected via qRT‒PCR, which was consistent with the RNA‒seq data. Here, we experimentally demonstrate for the first time that knockout of the WD40 domain of ATG16L1 enhances FMDV replication by downregulation innate immune factors. In addition, this result also indicates non-canonical autophagy inhibits FMDV replication. In total, our results play an essential role in regulating the replication level of FMDV and providing new insights into virus-host interactions and potential antiviral strategies.

Coordinate-based data analysis of the accuracy of five intraoral scanners for scanning completely dentate and partially edentulous mandibular arches.

STATEMENT OF PROBLEM: Current methods for assessing the accuracy of intraoral scanners (IOSs) that reduce errors and provide comprehensive data compared with previous methods are lacking. PURPOSE: The purpose of this in vitro study was to present a coordinate-based data analysis method to compare the accuracy of 5 IOSs for scanning completely dentate and partially edentulous casts. MATERIAL AND METHODS: Reference scans of 2 complete arch casts (completely and partially dentate) were digitized using a high-precision laboratory scanner (Ceramill Map 600). Each cast was scanned 10 times each using 5 IOSs (3Shape TRIOS 3, Planmeca Emerald, iTero Element 5D, Medit i500, and Shining Aoralscan 3). The dataset of all 10 test groups was analyzed by using a reverse engineering software program (Geomagic Wrap). Each test cast was aligned with the reference cast by 3-dimensional (3D) superimposition to determine the translation and rotation along the x-, y-, and z-axes. The dataset was analyzed using the Kruskal-Wallis and post hoc Bonferroni tests (α=.05). RESULTS: Significant differences were observed in all parameters among all scanners when scanning the same cast (P<.05). Significant differences were observed in at least 1 parameter for all scanners, except Element 5D after scanning different casts using the same scanner. Deviations in the test data generally relocated toward the mesial, buccal, and apical sides, and the casts were almost always rotated clockwise around the y-axis and counterclockwise around the z-axis. For the completely dentate cast, among all IOSs, Element 5D demonstrated the highest accuracy in most of the measured parameters, specifically in the y-axis translation (0.06[0.07] mm), z-axis translation (0.08[0.05] mm), and y-axis rotation (0.21[0.16] degree) (P<.05). For the partially edentulous cast, Element 5D displayed higher accuracy in most of the measured parameters, including the x-axis translation (0.11[0.14] mm) and z-axis rotation (0.12[0.18] degree) (P<.05). Emerald also displayed higher accuracy in most of the measured parameters, including the y-axis translation (0.05[0.08] mm) and y-axis rotation (0.14[0.12] degree) (P<.05). Element 5D exhibited no difference in the scanning accuracy between the 2 types of casts (P>.05). CONCLUSIONS: Element 5D offered a high level of accuracy and was an appropriate scanner for both situations. The method presented in this study provides a good assessment of accuracy deviations in complete arch scans using 3D coordinate-based data analysis.

Structural characterization and biological activities of a new polysaccharide isolated from Morchella Sextelata.

Morchella is the famous medicinal fungi in the ascomycetes. In this study, a new water-soluble polysaccharide (MSP-3-1) with an average molecular weight of 2.35 × 107 Da was extracted and purified from fruiting bodies of cultivated M. Sextelata. The structural characterization and biological activities of purified polysaccharide was further investigated. The results indicated that MSP-3-1 was mainly a α-glucan, mainly consisting of mannose (Man), glucose (Glc) and galactose (Gal) in a ratio of 5.10: 91.39: 3.51. Its surface morphology exhibited irregular lamellar structures with small voids. And the particle size analysis showed that MSP-3-1 was the homogeneous nanoparticle in water solution. Furthermore, the antioxidant activity analysis showed that MSP-3-1 possessed certain scavenging activity against hydroxyl radicals, DPPH radicals and ABTS radicals in a dose-dependent manner. Immunological tests suggested that MSP-3-1 could significantly promote the proliferation, phagocytosis and nitric oxide (NO) production of macrophage RAW264.7. Thus, our results will provide a theoretical basis for the development and utilization of Morchella Sextelata polysaccharides as an immunmodulatory component in functional foods.

Serum vaspin as a predictor of severity and prognosis in acute ischemic stroke patients.

Objective: The influence of vaspin on vascular health had been investigated, yielding conflicting results. This study is intended to investigate the relation between vaspin and stroke severity and stroke outcome in a cohort Chinese patient with acute ischemic stroke (AIS).Methods: This was a prospective single-center observational study in Xinxiang, China. From 1 July 2017 to 30 November 2019, all patients with first-ever AIS were consecutively included. Serum levels of vaspin, stroke severity at (assessed by NIHSS score) admission and functional outcome (assessed by modified Rankin Scale (mRS)) at discharge were recorded. Multivariate analyses were assessed using logistic regression models.Results: Finally, 340 patients with AIS were included. The median age of those patients was 65 (interquartile range [IQR], 56-74) years and 61.8% were men. At admission, 88 patients (25.9%) experienced severe stroke (NIHSS>10) and serum levels of vaspin (median [IQR]: 0.72[0.48-0.90]ng/ml) in those patients were significantly lower than in those mild(0.92[0.70-1.19]ng/ml) and moderate stroke (0.93[0.63-1.21]ng/ml). At discharge, 113 patients (33.2%) experienced poor functional outcome (mRS >2) and vaspin serum levels in those patients were lower as compared with patients who experienced good outcome (0.71[0.45-0.98] vs. 0.91[0.71-1.19]ng/ml). In multivariate analyses, lower level of vaspin (< median) was associated with a 2.5-fold (odds ratio [OR] 2.46; 95% confidence interval [CI]: 1.75-4.45) increased risk for severe stroke and a 2.1-fold (2.03; 1.42-3.58) increased risk for poor outcome.Conclusion: In conclusion, reduced serum levels of vaspin at admission are significantly related to stroke severity and prognosis, which illustrates a predictive role of reduced vaspin in ischemic stroke.

Evaluation of the accuracy of seven intraoral scanners for the full dentate and partially edentulous complete-arch mandibular casts: An in vitro comparison.

Statement of problem: Intraoral scanners (IOSs) are widely used in dentistry, providing high accuracy in short-range scanning. Nevertheless, when scanning the full dental arch, it remains a challenge. Furthermore, there is a lack of studies reporting the differences in scan accuracy between dental arches with large-span mucosal areas and fully dentate casts or optimal IOS selection for different dental statuses. Purpose: This study aimed to evaluate the accuracy and scanning time of different IOSs for full dentate (FD) and partially edentulous (PE) casts with missing teeth in the #34-#44 range and to determine the IOSs with the optimal clinical adaptability and scanning accuracy for different complete-arch casts. Material and methods: Reference scans of two complete-arch (FD and PE) casts were obtained using a laboratory scanner (Ceramill Map 600). Subsequently, the same casts were scanned ten times each by seven IOSs (3Shape Trios 3, CS3600, Planmeca Emerald, iTero Element 5D, Medit i500, BAMBOO B1, and Shining Aoralscan 3), and the scanning time was recorded. The test data were superimposed on the reference scans for the selected areas, and three-dimensional deviations between the reference and test casts (trueness), and between test casts (precision) were determined using reverse engineering software (Geomagic Wrap). The dataset was analyzed using a two-factor analysis of variance with post-hoc Bonferroni tests. Results: Two-factor analysis of variance revealed significant differences in accuracy and scanning time for different casts (P < 0.001) and IOSs (P < 0.001). For the FD cast, the i500 (0.35 ± 0.11 mm trueness) and CS3600 (0.23 ± 0.12 mm precision) performed worse than the remaining scanners. For the PE cast, the BAMBOO B1(0.89 ± 0.58 mm trueness; 0.88 ± 0.48 mm precision) performed worse than the remaining scanners. There were no differences in the accuracy of scanning between the Element 5D and Emerald for both cast types. However, the scanning time differed significantly between the different IOSs (P < 0.001). Regardless of the cast type, the fastest and slowest scans were performed by the Trios3 and CS3600 scanners respectively. Conclusions: The accuracy and scanning time differed between the different IOSs and types of complete-arch casts.

Prognostic Pathways Guide Drug Indications in Pan-Cancers.

Pathway-level analysis is a powerful approach enabling the interpretation of post-genomic data at a higher level than that of individual molecules. Molecular-targeted therapy focusing on cascade signaling pathways has become a new paradigm in anticancer therapy, instead of a single protein. However, the approaches to narrowing down the long list of biological pathways are limited. Here, we proposed a strategy for in silico Drug Prescription on biological pathways across pan-Cancers (CDP), by connecting drugs to candidate pathways. Applying on a list of 120 traditional Chinese medicines (TCM), we especially identified the "TCM-pathways-cancers" triplet and constructed it into a heterogeneous network across pan-cancers. Applying them into TCMs, the computational prescribing methods deepened the understanding of the efficacy of TCM at the molecular level. Further applying them into Western medicines, CDP could promote drug reposition avoiding time-consuming developments of new drugs.

[Study on oriented differentiation of bone marrow mesenchymal stem cells by fibroblast in rat uterine ligament with mechanical stretch].

OBJECTIVE: To investigate the effect on the differentiation of bone marrow mesenchymal stem cells (BMSC) with non-contact co-culture with mechanical stimulated ligament fibroblasts. METHODS: A cyclic 10% uniaxia strain at 1 Hz was applied on rat pelvic ligament fibroblasts, then were co-cultured with BMSC for 3, 6 and 12 days in non-contact condition. The protein expression of collagen I, III in BMSC were detected by SP method and revealed by the mean gray value. The mRNA expressions of collagens type I and type III in the BMSCs were measured with real-time (RT)-PCR, and the results were indicated by the ratio between the mRNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). RESULTS: (1) Protein expression: after 3 days co-culture with pelvic ligament fibroblasts, expression of collagen I and III in BMSC are 82.4±3.4 and 76.8±2.5. When compared with 80.2±2.6 and 74.6±1.1 in BMSC without co-culture, there was no significant difference (P>0.05). After 6 days co-culture with pelvic ligament fibroblasts, the expression of collagen I and III of 126.6±2.2 and 118.6±1.4 in BMSC were significantly higher than 82.7±3.0 and 76.2±1.3 in BMSC without co-culture (P<0.05). Similarly, after 12 days co-culture with pelvic ligament fibroblasts, the expression of collagen I and III of 135.3±3.4 and 128.7±2.6 in BMSC were significantly higher than 86.6±1.3 and 81.8±1.4 in BMSC without co-culture (P<0.05). (2) mRNA expression: after 3 days co-culture with pelvic ligament fibroblasts, the mRNA expression of type I and type III collagens in BMSC are 2.10±0.20 and 1.20±0.30. When compared with mRNA expression of 2.01±0.12 and 1.13±0.21 in BMSC without co-culture, no significant difference were observed (P>0.05). After 6 days co-culture with pelvic ligament fibroblasts, the mRNA expressions of type I and type III collagens mRNA were 5.60±0.21 and 2.61±0.20, which were significantly higher than 3.70±0.33 and 1.82±0.14 in BMSC without coculture (P<0.05). After 12 days co-culture with pelvic ligament fibroblasts, the mRNA expressions of type I and type III collagens of 5.91±0.31 and 2.92±0.23 were significantly higher than 4.04±0.21 and 2.04±0.13 in BMSC without co-culture (P<0.05). CONCLUSION: Non-contact co-culture with mechanical stretch stimulated ligament fibroblasts, it might promote synthesis of types Iand III collagen in rat BMSCs and induced BMSC differentiated into pelvic ligament fibroblasts.

Clinical Features of Trousseau Syndrome With Cerebral Infarction as the Initial Manifestation.

OBJECTIVE: There are few reports of Trousseau syndrome with cerebral infarction as the initial manifestation before the discovery of the tumor, which is often missed and misdiagnosed, and there is no unified therapy. To explore the clinical features of the Trousseau syndrome and, among those features, the risk factors for cerebral infarction as the initial manifestation. METHODS: This was a retrospective study of 416 consecutive patients with cerebral infarction and malignant tumor admitted at The First Affiliated Hospital of Xinxiang Medical University between January 2015 and December 2017. The patients were grouped as: (1) cerebral infarction as the initial manifestation; and (2) tumor as the initial manifestation. A multivariable logistic regression analysis was used to analyze the relationship between the clinical features (age, sex, characteristics of the infarction, characteristics of the tumors, treatments, depression, coagulopathy, The National Institute of Health stroke scale score, platelet count, red cell count, hemoglobin, atherosclerosis, and coagulation parameters) and the hypercoagulable state. RESULTS: A total of 416 patients met the criteria were included: 212 (51.0%) in the group with cerebral infarction as the initial manifestation and 204 (49.0%) in the group with tumor as the initial manifestation. The multivariable analysis showed that metastatic cancer (odds ratio=2.517; 95% confidence interval, 1.193-5.311; P=0.015) and depressive state (odds ratio=3.158; 95% confidence interval, 1.522-6.551; P=0.002) were independently associated with the Trousseau syndrome with cerebral infarction as the main manifestation. CONCLUSIONS: Trousseau syndrome with cerebral infarction as the initial manifestation was associated with metastatic cancer and depressive state. There was no difference in coagulation status between the 2 groups.

Plasma levels of ceramides relate to ischemic stroke risk and clinical severity.

Recent studies have suggested that specific plasma ceramides are independently associated with atherosclerosis and cardiovascular diseases, but it is currently unknown whether plasma ceramide levels are associated with ischemic stroke. Here, we examined whether ceramides were associated with both ischemic stroke risk and clinical severity at admission. We measured three previously identified high-risk plasma ceramide molecules [Cer(d18:1/16:0), Cer(d18:1/22:0), and Cer(d18:1/24:0)] in 202 patients with acute ischemic stroke and 202 age and sex matched control cases. Plasma ceramides levels were measured by a targeted liquid chromatography-tandem mass spectrometry assay at baseline. The median age of the 202 stroke patients was 66 (interquartile range [IQR], 58-75) years and 54.0 % were men. Plasma levels of C16:0, C22:0, and C24:0 ceramides in stroke patients were significantly higher than in those control cases (P < 0.001, all). In multivariate logistic regression analysis adjusted for other risk factors, higher levels of C16:0, C22:0, and C24:0 ceramides were associated with higher risk of ischemic stroke (odd ratio [OR] for one IQR increase: 2.15[1.42-2.99]; 2.90[2.13-4.01] and 1.29[1.10-1.69]; respectively). At admission, 103 patients (51.0 %) had a minor stroke (NIHSS < 6). In these patients, plasma levels of C16:0, C22:0, and C24:0 ceramides were lower than that observed in patients with moderate-to-high clinical severity (P < 0.001, all). In multivariate logistic regression analysis adjusted for other risk factors, higher levels of C16:0, C22:0, and C24:0 ceramides were associated with higher risk of moderate-to-high stroke (OR for one IQR increase: 2.96 [2.05-4.22], 3.03 [2.01-4.25] and 1.72 [1.25-3.31], respectively). An elevated plasma levels of ceramides were predictors of both risk and severity at admission in ischemic stroke patients. The underlying mechanisms of these associations remain to be investigated.

Choledochoscopic lithotripsy is a useful adjunct to laparoscopic common bile duct exploration for hepatolithiasis: a cohort study.

BACKGROUND: This study aims to investigate the role of combining choledochoscopic lithotripsy with laparoscopic common bile duct exploration for hepatolithiasis in patients who are not suitable for hepatectomy. METHODS: From March 2009 to March 2013, 86 patients with hepatolithiasis irrespective of whether they underwent a choledochoscopic plasma shock wave lithotripsy or not were analyzed. RESULTS: Sixty-two patients underwent lithotripsy and 24 patients underwent basket lithoextraction intraoperatively. Plasma shock wave lithotripsy did not lengthen the operating time, but decreased the postoperative residual stone rate and reduced the frequency of postoperative choledochoscopic lithotomy for patients with remnant stones. The overall final stone clearance rate was 98.8%. During a mean follow-up of 26.2 months, recurrent stones and cholangiocarcinoma developed in 1 patient, respectively. CONCLUSION: Laparoscopic common bile duct exploration combined with choledochoscopic lithotripsy is a definitive procedure for hepatolithiasis in patients who are not candidates for hepatectomy.