The Reliability of Three-Dimensional Landmark-Based Craniomaxillofacial and Airway Cephalometric Analysis.

Cephalometric analysis is a standard diagnostic tool in orthodontics and craniofacial surgery. Today, as conventional 2D cephalometry is limited and susceptible to analysis bias, a more reliable and user-friendly three-dimensional system that includes hard tissue, soft tissue, and airways is demanded in clinical practice. We launched our study to develop such a system based on CT data and landmarks. This study aims to determine whether the data labeled through our process is highly qualified and whether the soft tissue and airway data derived from CT scans are reliable. We enrolled 15 patients (seven males, eight females, 26.47 ± 3.44 years old) diagnosed with either non-syndromic dento-maxillofacial deformities or OSDB in this study to evaluate the intra- and inter-examiner reliability of our system. A total of 126 landmarks were adopted and divided into five sets by region: 28 cranial points, 25 mandibular points, 20 teeth points, 48 soft tissue points, and 6 airway points. All the landmarks were labeled by two experienced clinical practitioners, either of whom had labeled all the data twice at least one month apart. Furthermore, 78 parameters of three sets were calculated in this study: 42 skeletal parameters (23 angular and 19 linear), 27 soft tissue parameters (9 angular and 18 linear), and 9 upper airway parameters (2 linear, 4 areal, and 3 voluminal). Intraclass correlation coefficient (ICC) was used to evaluate the inter-examiner and intra-examiner reliability of landmark coordinate values and measurement parameters. The overwhelming majority of the landmarks showed excellent intra- and inter-examiner reliability. For skeletal parameters, angular parameters indicated better reliability, while linear parameters performed better for soft tissue parameters. The intra- and inter-examiner ICCs of airway parameters referred to excellent reliability. In summary, the data labeled through our process are qualified, and the soft tissue and airway data derived from CT scans are reliable. Landmarks that are not commonly used in clinical practice may require additional attention while labeling as they are prone to poor reliability. Measurement parameters with values close to 0 tend to have low reliability. We believe this three-dimensional cephalometric system would reach clinical application.

Genome Characterization and Phylogenetic Analysis of the First Bovine Rhinitis B Virus Isolate in China.

Bovine rhinitis B virus (BRBV) is an emerging viral species in the genus Aphthovirus, family Picornaviridae. Studies suggested that BRBV was considered a potential etiological agent of bovine respiratory disease complex (BRDC). BRBV has been reported in the United States, Sweden, Canada, Japan, and Mexico. However, little information of BRBV was available in China. In this study, we performed viral metagenomic analysis in a calf with respiratory disease. The results showed high abundance (3.85) of BRBV nucleotide and 248 mapped reads in calf samples. Online BLASTn analysis showed that three contigs of those had the highest nucleotide similarity (95%) with one Swedish BRBV isolate (BRBV_SWE1, GenBank accession no. KY432299). To identify the genome characterization of the Chinese BRBV isolate (designated CHN1), six couples of overlapping RT-PCR primers were designed according to genome sequences of BRBV_SWE1. Through gene cloning and splicing, we obtained the genome information of CHN1, possessing 7,465 nucleotides (46.6% G+C). Although CHN1 had the highest nucleotide similarity (95.1%) with BRBV_SWE1, one 11-nucleotide (ACATTTGTTGT) deletion occurred in the 5' untranslated region compared to SWE1. Phylogenetic analysis showed that CHN1 clustered together with BRBV_SWE1, and far from other BRBV isolates. This study recorded the first discovery of BRBV infection in China. Further investigation should be made in order to evaluate the infection status and epidemiological significance of BRBV in China.

Development of a Real-Time Quantitative RT-PCR Assay for Detection of Bovine Rhinitis B Virus.

Bovine rhinitis B virus (BRBV) has been frequently identified in cattle diagnosed with bovine respiratory disease complex (BRDC) in recent years, suggesting its potential contribution to BRDC. The goal of this study was to develop a TaqMan-based real-time quantitative RT-PCR assay for efficient BRBV detection. A pair of primers and a probe were designed based on the 3D gene of the BRBV genome. The assay was specific for BRBV and able to exclude bovine rhinitis A virus, foot-and-mouth disease virus and Senecavirus A. The limit of detection of the assay was 4.46 copies per reaction. A standard curve was plotted, with a coefficient of determination of 0.999 in the concentration range of 100-108 copies/µl. The reproducibility of the assay was acceptable, with the standard deviations of cycle threshold values lower than 1.00 in both intra- and inter-assay. Of 200 samples collected from 150 head of cattle in recent years in China, 11% (22/200) of the samples tested positive in the assay, i.e., 4.6% (7/150) of the cattle were BRBV positive. This study provides an efficient diagnostic tool for the epidemiological investigations of BRBV.

A Novel Technique for Constructing Infectious Cloning of Type 3 Porcine Circovirus.

Porcine circovirus type 3 (PCV3), which currently lacks effective preventive measures, has caused tremendous economic losses to the pig husbandry. Obtaining the strain of PCV3 is the key to preparing related vaccines and developing corresponding antiviral drugs. In this study, according to the linear sequence of PCV3 DNA published on GenBank, the sequence was rearranged with SnapGene gene-editing software, and after rearrangement, the HindIII restriction endonuclease site was added to the end of the linear DNA, so that both ends have the same restriction endonuclease site. On this basis, the rearranged linear DNA is obtained by gene synthesis, PCR amplification, DNA purification, etc., and is digested and connected in vitro to obtain cyclized DNA. PCV3 infectious clones were obtained by transfecting 3D4/21 cell lines. The obtained PCV3 was identified by PCR, Western blotting, and indirect immunofluorescence tests. The results showed that this study successfully obtained the strain of PCV3 in vitro. To further evaluate the pathogenicity of the obtained PCV3 infectious clones, this study established an animal model of Kunming mice infected with PCV3. The results of RT-PCR, Western blotting and immunohistochemistry showed that PCV3 can infect myocardium and alveoli of Kunming mice, but no PCV3 was detected in other tissues. The above studies indicate that PCV3 circular DNA can be used to construct PCV3 infectious clones. This research will provide a new method for the construction of circular DNA viruses and lay the foundation for the research and pathogenesis of PCV3 vaccine.

Intracellular lipid dysregulation interferes with leukocyte function in the ovaries of meat-type hens under unrestricted feed intake.

Meat-type Red-feather country hens fed ad libitum (AD-hens) exhibit obesity-associated morbidities and a number of ovarian irregularities. Leukocyte participations in ovarian activities are unstudied in AD-hens. In contrast to feed-restricted hens (R-hens), ovulatory process of the F1 follicle appeared delayed in AD-hens in association with reduced F1 follicle progesterone content, gelatinase A (MMP-2) and collagenase-3 (MMP-13) activities coincident with elevated IL-1ß and no production (P<0.05), and increased leukocyte infiltration of inflamed necrotic follicle walls. Extracts of AD-hen F1 follicle walls induced greater leukocyte migration than extracts from F1 follicle wall extracts of R-hens (P<0.05). Co-cultures of granulosa cells with increasing numbers of leukocytes from either AD-hens or R-hens exhibited dose dependent reductions in progesterone production and increases in cell death. AD-hen leukocytes were less proapoptotic than their R counterparts (P<0.05). Granulosa MMP-13 and MMP-2 activities were also suppressed in the co-cultures with heterophils or monocytes in a dose-dependent manner (P<0.05). AD heterophils and R monocytes had a greater inhibitory effect on MMP activities in the co-cultures than their respective counterparts (P<0.05). Both basal and LPS-induced IL-1ß secretion and MMP-22 or MMP-2 activities in freshly isolated AD-hen leukocytes were reduced (P<0.05). Exposure of AD or R leukocytes to 0.5mM palmitate impaired IL-1ß secretion and MMP-22 or MMP-2 activity. Inhibition of ceramide synthesis with FB1 and ROS production with n-MPG scavenging rescued MMP activity and IL-1ß production in palmitate treated heterophils, but exacerbated monocyte suppression. These latter findings suggest that intracellular lipid dysregulation in leukocytes contributes to ovarian dysfunction in AD-hens.

[Accuracy and reproducibility of intraoral scanning in vivo].

PURPOSE: To evaluate the accuracy and reproducibility of intraoral scanning system by comparing linear measurements obtained from conventional models and digital models. METHODS: Both conventional models (control group) and intraoral scanning (experimental group) of 5 volunteers were made. After taking silicon impressions, the volunteers' oral cavities were scanned directly by Trios Ortho (3 Shape) at 8a.m., 13p.m., 18p.m. and repeated for 15 days. The gypsum models were measured by digital caliper and regarded as golden standard. The intraoral scanning files were measured by Simplant software. A well-trained examiner measured them in random order at 8a.m., 13p.m. and 18p.m. The measurements were repeated for 15 times. Datasets obtained from different groups were compared for accuracy via paired t test. A one-way analysis of variance (ANOVA) was implemented to compare differences within experimental groups for reproducibility using SAS 11.0 software package. RESULTS: There was a significant linear difference between gypsum model and intraoral scanning digital model in the parameter of 23L from volunteer V1 (P<0.01). In terms of accuracy and reproducibility, no significant difference was found (P>0.05). CONCLUSIONS: The accuracy and reproducibility of intraoral scanning on whole arches are quite acceptable in this in vivo study.

Feed intake alters immune cell functions and ovarian infiltration in broiler hens: implications for reproductive performance.

Leukocytes are known to participate in ovarian activities in several species, but there is a surprising lack of information for the common chicken. Broiler hens consuming feed ad libitum (AL) exhibit a number of ovarian irregularities, but leukocyte functions are unstudied. In contrast to feed-restricted (R) hens, AL feeding for 7 wk significantly reduced egg production and clutch length while increasing pause length and atretic follicle numbers (P < 0.05). Granulosa cells from F1 follicles of AL hens contained less progesterone, and follicle walls were thicker with loose fibrous morphology and had less collagenase-3-like gelatinolytic activity but more IL-1beta (P < 0.05) production, suggestive of slower maturation in ovulatory process and inflamed necrosis. Interestingly, while highly infiltrated with immune cells, particularly heterophils, IL-1beta, MMP-22-like, and gelatinase A activities were reduced in AL hen peripheral heterophils and monocytes (P < 0.05); however, AL monocytes showed an increase in phagocytosis rate (P < 0.05). Generation of reactive oxygen intermediates was also suppressed in AL heterophils but increased in AL monocytes (P < 0.05). In contrast to leukocyte-free control, both AL and R heterophils and monocytes suppressed progesterone production and increased cell death in a dose-dependent manner when coincubated with granulosa cells at different ratios (P < 0.05). AL monocytes suppressed progesterone production more, but AL heterophils were less proapoptotic when compared to their R counterparts (P < 0.05). Alterations of cellular ceramide content (P < 0.05) corresponded to the discrepancy between heterophil and monocyte functionality. In conclusion, leukocyte dysfunction contributes to impaired ovarian activities of overfed broiler hens.