[The accuracy and feasibility study of freehand pedicle screw insertion for subaxial cervical spine assisted with safe core-referred technique].

Objectives: To construct the "safe core" of the pedicle screw trajectory using CT imaging data of the subaxial cervical spine in adults, and to assess the accuracy and feasibility of the pedicle screw insertion assisted with the "safe core-referred technique" for subaxial cervical spine with a cadaver specimen study. Methods: This is an experimental study. From January 2015 to March 2020,60 adults' CT images data of the cervical spine were collected from the database of the First Affiliated Hospital of Gannan Medical University,and were imported into Mimics 20.0 software. Virtual cervical pedicle trajectory and safe core were constructed according to the self-designed "virtual construction method of pedicle in the subaxial cervical spine". The success rate of the construction and the spatial position data of the virtual safe core of was recorded,including the distance between the safe core and the tangent line of the upper and lower outer edge of Luschka's joint on coronal plane,and the distance between the safe core and the posterior edge of the vertebral body on sagittal plane.The 3.5 mm column was used to simulate the pedicle screw placement,using the safe core as the only hub in pedicle screw trajectory.The length of the anterior pedicle screw trajectory,the interval of the abductive angle of the pedicle screw in axial plane, and the projection area of the entry area on periapical radiograph was calculated.In addition,8 adult cervical cadaver specimens were collected for the pedicle screw insertion experiment.The left side group used the "safe core-referred technique" for pedicle screw insertion,while the right side group used the Abumi method for pedicle screw insertion.The accuracy of pedicle screw placement was verified by CT scan.The difference between the accuracy of subjective judgment based on X-ray monitoring of operator and the actual accuracy of pedicle screw insertion verified by CT scan was compared between the two groups.The chi-square test was used to compare the intergroup data. Results: The total success rate of the virtual construction method for the safe core of the subaxial cervical spine was 97.0% (291/300); The distance between the safe core and the tangent line of the upper and lower outer edge of Luschka's joint on coronal plane was (M(IQR)) 0.91 (0.98) mm (range: 0 to 1.85 mm);The distance between the safe core and the posterior wall on the sagittal plane of the vertebral body was (2.01±0.86) mm (range: 0.67 to 3.53 mm). The distance (anterior pedicle screw trajectory) from the posterior cortex to the central point of the safe core was (11.58±1.00)mm (range: 8.27 to 14.93 mm).The projection area of the entry point on the coronal plane was (36.18±11.67) mm2 (range: 13.38 to 83.11 mm2). Pedicle screw insertion experiment in cervical cadaver specimen showed the rate of intraoperative correction of the pedicle screw trajectory was 7.5% (3/40) in the experimental group and 12.5% (5/40) in the control group (χ2=0.139,P=0.709). The operator 's correct rate of subjective judgment on CT in the stage of pedicle screw trajectory preparation was 100% (40/40) in the experimental group and 82.5% (33/40) in the control group, the difference was statistically significant (χ2=5.638,P=0.018). The actual correct rate of CT verification in the stage of pedicle screw insertion was 100% (40/40) in the experimental group and 90.0% (36/40) in the control group, the difference was statistically significant (χ2=2.368,P=0.124); The operator 's correct rate of subjective judgment in the stage of pedicle screw insertion completion was 100% (83/83) in the experimental group and 92.9% (79/85) in the control group (χ2=4.199,P=0.040). Conclusions: The virtual safe-core of subaxial cervical spine can be use as a reliable anatomical fluoroscopy landmark for freehand pedicle screw insertion."Safe core-referred technique" can improve the accuracy rate of the operator's subjective judgment on the intraoperative fluoroscopy monitoring,and hence improve the accuracy of freehand pedicle screw insertion technology for subaxial cervical spine. And it still needs to be further verified in clinical practice.

[Establishment and application of a multilocus sequence typing assay for Corynebacterium striatum].

Objective: To establish a multilocus sequence typing (MLST) assay for Corynebacterium (C.) striatum, explore the population structure and evolution relationship of clinical isolates of C. striatum. Methods: Seven housekeeping genes (gyrA, gyrB, hsp65, sodA, secA1, rpoB, 16S rRNA) were amplified with PCR by using self-designed specific primers and sequenced. Then, the sequences were assembled with software SeqMan. The gene diversity and gene recombination characteristics were evaluated by using software DnaSP 5.10.01 and Splits tree 4.14.2. The phylogenetic tree and the minimum spanning tree were constructed based on the sequence types (ST) characteristics by using software MEGA 7.0.14 and BioNumerics, respectively. In addition, the genetic evolutionary relationship among STs were analyzed by using software eBURST 3.0. Results: The expected amplification products of seven sites selected in all the test strains were obtained. Splits tree showed that the clustering of all C. striatum strains was consistent, suggesting that gene recombination is the potential driving force for the evolution of C. striatum. All of the 344 C.striatum strains were divided into 72 STs by MLST and 85.7% of the strains formed clonal complexes. CC19 was the predominant clonal complex, whereas ST16 in the clonal complex was detected in the most strains. ST had a certain geographic clustering and a certain correlation with the isolation time. Conclusions: C. striatum showed high genetic diversity in China and CC19 was the predominant clonal complex. The MLST assay established in this study can be used for the typing of C. striatum, but further improvement is needed.

Impact of astrocyte and lymphocyte interactions on the blood-brain barrier in multiple sclerosis.

OBJECTIVE: This study was designed to investigate the impact of astrocyte and lymphocyte (LC) interactions in the blood-brain barrier (BBB) on the pathogenesis of multiple sclerosis (MS). METHODS: Primary rat brain microvascular endothelial cells (rBMECs) and astrocytes isolated from Sprague-Dawley rats were used to establish in vitro BBB models. Transendothelial electrical resistance (TEER) and permeability were compared between rBMEC monocultures and rBMEC/astrocyte co-cultures to evaluate the validity of each as a BBB cell model. rBMEC/LC co-cultures and rBMEC/astrocyte/LC tri-cultures were established to evaluate inflammatory responses in MS by measuring the gene expression levels of nerve growth factor (NGF), matrix metalloproteinase 2 (MMP-2), matrix metalloproteinase 9 (MMP-9), interleukin 17 (IL-17), interferon γ (IFN-γ), and forkhead box P3 (Foxp3). RESULTS: The rBMEC/astrocyte co-cultures exhibited higher TEER values and lower lymphocyte permeabilities than those of rBMEC monocultures. Compared to the rBMEC mono-cultures, the rBMEC/astrocyte/LC tri-cultures showed significantly decreased NGF, IL-17, and IFN-γ and increased MMP-2 and Foxp3 expression. Furthermore, the tri-cultures exhibited decreased NGF, IL-17, and IFN-γ expression compared to the rBMEC/astrocyte co-cultures, and increased MMP-2 expression compared to that shown by the rBMEC/LC co-cultures. MMP-9 expression did not vary significantly between the four established BBB cell models. CONCLUSION: These results suggest that the synergistic effect between astrocytes and LCs may increase the expression of MMP-2 and decrease that of IL-17 and IFN-γ at the BBB. Furthermore, the use of rBMEC/astrocytes/LC tri-cultures enabled us to test the synergistic effect between astrocytes and LCs and their roles in MS pathogenesis.

The prognostic roles of long non-coding RNA SNHG17 in the patients with gastric cancer.

OBJECTIVE: Dysregulated long non-coding RNAs (lncRNAs) might exert critical roles in pathways associated with gastric cancer (GC) development. Long non-coding RNA (SNHG17), a newly identified lncRNA, has been reported to be dysregulated in several tumors. Our present study aimed to explore the expression level of SNHG17 in patients with GC and investigate the relationship between the SNHG17 level and the prognosis of GC patients. PATIENTS AND METHODS: The expression levels of SNHG17 were detected by Real-time-quantitative Polymerase Chain Reaction (RT-qPCR). The chi-square test was performed to investigate the associations between SNHG17 expression and the clinical features of GC patients. The prognostic value of SNHG17 was demonstrated using Kaplan-Meier method and multivariate Cox regression analysis. RESULTS: We showed that SNHG17 was significantly up-regulated in GC tissues compared with adjacent normal gastric tissues (p<0.01) and higher expression of SNHG17 was observed in advanced GC tissues. The results by analyzing the clinicopathological features showed that overexpression of SNHG17 expression was associated with advanced TNM stage (p=0.006), positively lymph node metastasis (p=0.006) and positively distant metastasis p=0.024). Moreover, clinical assays revealed that patients with high SNHG17 expression levels had a significantly shorter overall survival (OS) (p=0.0034) and progression-free survival (PFS) (p=0.0002) than those expressing lower. Finally, multivariate assays showed that SNHG17 was an independent prognostic marker for both OS and PFS of patients with GC. CONCLUSIONS: Our findings indicated that SNHG17 was a novel molecule involved in GC progression, providing a potential prognostic biomarker for GC patients.

Mapping quantitative trait loci for cytokines in the pig.

Increased disease resistance through improved general immune capacity would be beneficial for the welfare and productivity of farm animals. Cytokines are essential diagnostic parameters in veterinary practice. To identify quantitative trait loci (QTL) for cytokine levels in serum in the pig, Interferon-gamma (IFN-γ) and Interleukin 10 (IL-10) levels and the ratio of IFN-γ to IL-10 were measured in a composite pig population, before and after challenge with modified live CSF (classical swine fever) vaccine. Through interval mapping using the variance component approach and the permutation test, 11 QTL (five for IFN-γ, two for IL-10 and four for the ratio of IFN-γ to IL-10) with significance levels of P < 0.10 were identified, of which five were significant at the P < 0.05 level. The most significant QTL (P < 0.01) was found on chromosome 16, with effect on the ratio of IFN-γ to IL-10. Within these QTL regions, a number of known genes were revealed and their potential relationships to the studied traits were discussed. Some of these genes may serve as candidate genes for these traits in swine.

Association analysis of polymorphisms of porcine LMP2 and LMP7 genes with haematological traits.

Low molecular weight polypeptides 2 (LMP2) and low molecular weight polypeptides 7 (LMP7) are located within the major histocompatibility complex and have been associated with autoimmune disease. In this study, polymorphisms of porcine LMP2 and LMP7 genes were analyzed by PCR-SSCP and DNA sequencing methods. Four SNPs (DQ659151:g.2115T>C; DQ659151:g.4343A>G; DQ872631:g.1232C>G; DQ872631:g.2847C>T) were identified. Four SNPs of genes were analyzed for association with 22 haematological traits in Large White (n = 195), Landrace (n = 84) and Songliao Black (n = 86) pig population. Of all the 22 traits, seven were significant associated with the SNPs of LMP2/LMP7 gene (P < 0.05). They included white blood cell count (WBC) (P = 0.028), neutrophilic granulocyte count (GRAN) (P = 0.037), monocytes percentage (MO%) (P = 0.015), red blood cell (RBC) (P = 0.004), red blood cell volume distribution width (RDW) (P = 0.004), mean platelet volume (MPV) (P = 0.016) and CD4(+)CD8(+)% (P = 0.045). These results suggest LMP2/LMP7 gene should be regarded as molecular marker to estimate animal's immune status for their effects on hematological traits.