Objective:To investigate the anatomical safety and feasibility ofposterior occipitocervical fixation with atlan-tooccipital-clivus screw.Methods:Data of 60 patients who treated in the spinal department of our hospital with upper cervical computed tomographic scans from February 2017 to November 2019 were retrospectively collected. Occipitocervical infection, injury, tumor and deformity were excluded. The Mimics software was used to reconstruct the occiput, atlas and measure the anatomical parameters, including the height and width of the anterior edge of the clivus, the height and width of the middle part of the clivus, the thinnest distance of the soft tissue in front of the clivus, the anteroposterior diameter, transverse diameter, the angle of inside tilting in coronary plane of the occipital condyle, the distance from the hypoglossal canal to the atlantooccipital articular surface, the anteroposterior diameter and transverse diameter of the superior joint of atlas, the height of the lateral mass, and the height and transverse diameter of the inferior articular process of the superior atlas joint. The three-dimensional digital modeling was performed and the screw diameter of 3.5mm was simulated. 3-Matic software were used to measure the screw placement parameters, including the inside tilting angle in coronary plane of screw, and the angle of upper tilting in sagittal plane and length of screw. The atlanto-occipital junction was exposed at the rear of 8 cadavers. According to the above parameters, the titanium alloy screws with a diameter of 3.5 mm were transferred from the inferior articular process and posterior arch of the atlas to the clivus through the atlantooccipital. Finally, the screw path was cut along the nail path with a pendulum saw, and the track of the screw was observed to confirm the safety and effectiveness of the screw.Results:The leading edge height and width of male clivus was 16.8±2.5 mm and 20.1±3.1 mm. The middle part of the clivus was 9.7±2.3 mm and 22.4±3.7 mm. The thinnest soft tissue in front of the clivus was 5.8±1.48 mm. The anteroposterior diameter of the occipital condyle was 19.1±1.9 mm, the transverse diameter was 12.6±2.0 mm, the inside tilting angle was 33.7°±4.5°, and the vertical distance from the lowest point of the neural tube to the articular surface of the occipital condyle was 9.6±1.1 mm. The height of the lateral mass of atlas was 12.9±2.4 mm, the anteroposterior diameter of the upper joint of atlas was 21.7±1.9 mm, and the transverse diameter was 11.7±1.4 mm. The width of the inferior facet was 14.9±1.4 mm and the height of the inferior facet was 5.7±0.85 mm. The distance from the screw entry point to the vertical line of the lateral mass migration midpoint was 2.5±0.6 mm; The distance from the screw entry point to the horizontal line of the midpoint was 2.3±0.7 mm.The inside titling angle of screw was 18.4°±1.6°, the upper tilting angle was 55.6°±3.1°, the length of the screw track was 53.0±2.8 mm, the adjustment range of upper tilting angle was 15.0±2.8 mm, the adjustment range of inside tilting angle was 10.4±2.4 mm. The anatomical parameters of females were slightly smaller than those of males, and the difference was statistically significant, but there was no significant difference between left and right parameters. The screws of 8 specimens could be inserted safely and effectively.Conclusion:Atlan-tooccipital-clivus screw can be implanted without damaging the nerve and vascular structure, and it can be used as a choice for occipitocervical fixation.

Objective: To isolate and identify Prevotella nigrescens (P.nigrescens) in gingival crevicular fluid of patients with chronic periodontitis and to analyze its tumor-promoting role in esophageal squamous cell carcinoma (ESCC). Methods: Samples of gingival crevicular fluid were collected from patients with chronic periodontitis and cultured on GAM agar medium under anaerobic conditions. Black colonies on GAM agar plates were picked for subculture, and then the bacteria were isolated and purified. Bacterial identification was conducted using Gram staining and 16S rDNA sequencing analysis. The isolated bacterial species was used to infect ESCC NE6-T cells and the changes in biological characteristics of the infected cells were evaluated. Results: Multiple bacterial colonies were observed after anaerobic culturing of gingival crevicular fluid samples for 120 h on GAM agar plates. A single bacterial colony with pure black and smooth appearance was obtained from grey black bacterial colonies with streak method. It was a Gram-negative bacterium with bead-like shape. It showed 99.78% 16S rDNA sequence identity with P. nigrescens F0103 and was named P. nigrescens LY01. P. nigrescens LY01 infection promoted the in vitro proliferation, migration and invasion of NE6-T cells and the growth of subcutaneous xenograft in nude mice. In addition, it could also induce the upregulation of Ki67 and activation of p-STAT3. Conclusions P. nigrescens inhabiting in gingival sulcus might promote the progression of ESCC in human with chronic periodontitis.

Objective:To investigate the safety and efficacy of short-segment fixation covering the fractured vertebrae via posterior intermuscular approach plus percutaneous kyphoplasty (PKP) through the outer upper edge of the base of the fractured vertebral pedicle in the treatment of osteoporotic thoracolumbar burst fracture.Methods:A retrospective case series study was used to analyze the clinical data of 56 patients with osteoporotic thoracolumbar burst fracture admitted to Ningbo No.6 Hospital from January 2018 to February 2021, including 24 males and 32 females; aged 56-72 years [(63.5±4.6)years]. All patients underwent short-segment fixation covering the fractured vertebrae via posterior intermuscular approach combined with PKP through the outer upper edge of the base of the fractured vertebral pedicle. The operation time, intraoperative blood loss, hospitalization day and surgery-related complications were recorded. The visual analogue score (VAS) of back pain, ratios of the anterior, middle and posterior height of the fractured vertebrae and kyphotic Cobb angle were compared before operation, at postoperative 2 days and at the final follow-up.Results:All patients were followed up for 12-28 months [(14.5±2.2)months]. The operation time was 55-85 minutes [(62.0±12.1)minutes], intraoperative blood loss was 80-150 ml [(94.0±18.5)ml], and hospitalization day was 5-9 days [(7.4±1.1)days]. Based on CT examination at postoperative 2 days, there were 2 patients with paravertebral cement leakage, 2 with intervertebral space leakage and 1 with intracanal leakage, but none reported associated clinical symptoms. No implant failure or fractures of adjacent segments was detected during the follow-up period. The VAS was significantly decreased from preoperative (7.5±1.2)points to (3.2±0.8)points at postoperative 2 days ( P<0.01), and the score was further lowered to (2.2±0.8)points at the final follow-up when compared with that at postoperative 2 days ( P<0.01). The ratios of the anterior, middle and posterior height of the fractured vertebrae and kyphotic Cobb angle were significantly improved at postoperative 2 days [(89.5±13.2)%, (85.8±7.9)%, (89.5±9.0)% and (5.6±3.2)°] when compared with those before operation [(48.9±11.8)%, (61.9±11.9)%, (79.9±9.8)% and (26.3±5.6)°] (all P<0.01). Slight losses were observed in the ratios of the anterior, middle and posterior height of the fractured vertebrae and kyphotic Cobb angle at the final follow-up [(87.0±12.7)%, (82.1±7.8)%, (88.6±10.0)% and (5.4±3.2)°], but not significantly different from those at postoperative 2 days (all P>0.05). Conclusion:Short-segment fixation covering the fractured vertebrae via posterior intermuscular approach plus PKP through the outer upper edge of the base of the fractured vertebral pedicle can safely and effectively treat osteoporotic thoracolumbar burst fracture, for it can significantly improve back pain, restore the height of the fractured vertebrae and correct the kyphotic deformity.

Objective:To investigate the safety of posterior atlantoaxial transarticular screw combined with atlas pedicle screw fixation in Chinese.Methods:CT data of upper cervical spine in 48 patients were collected from Ningbo No.6 Hospital, including 26 males and 22 females aged 26-58 years [(37.3±13.5)years]. All CT data was transformed into 3D dimensional model and inserted with virtual screws by Mimics 19.0. Firstly, the vertical plane P 1 and the horizontal plane P 2 of the atlas were built in those atlantoaxial models, secondly the atlantoaxial transarticular screw S 0 was inserted by Margel method, and it's insertion point was located at 3 mm lateral and 2 mm cephalad in the C 2 inferior articular process. Finally, four atlas pedicle screws were inserted at the midline of atlas lateral mass. The four screws are inserted as follows. S 1: the screw was tangent to the lateral side of the S 0 or the medial of the atlas pedicle. S 2: the screw was tangent to the lateral wall of the atlas pedicle. S 3: the screw was tangent to the upper wall of the atlas pedicle. S 4: the screw was tangent to the inferior wall of the atlas pedicle. The angles between S 1, S 2 and P 1 as the camber angle, and the angle between S 3, S 4 and P 2 as the gantry angle were measured. Then the safety range of camber angle and gantry angle were calculated, and the screw length of S 1, S 2, S 3 and S 4 was measured. In all models, the camber angle and gantry angle of the screws were adjusted an interval of 2°, the number of successful cases was calculated and the success rate of insertion was calculated. Results:All 3D models were inserted successfully. The minimum value, maximum value and safety range of the camber angle of atlas pedicle screws were (-6.7±5.2)°, (10.4± 4.3)°, (17.1±3.7)°, respectively; and of the gantry angle were (-0.5±3.5)°, (11.0±5.8)°, (11.5±4.9)°, respectively. The length of screw placement was S 1: (31.1±2.4)mm, S 2: (28.3±2.5)mm, S 3: (29.2±3.8)mm, S 4: (29.6±3.0)mm, respectively ( P<0.05). When the camber angle was from -1°to 5°, the success rate of screw placement was 87.5%; when inserting with 2° camber angle, the success rate was 100%; when the gantry angle was 5°, the success rate was 93.8%. Conclusion:The posterior atlantoaxial transarticular screw combined with atlas pedicle screw can achieve satisfy insertion length and success rate.

Objective:To investigate the effects of Porphyromonas gingivalis ( P. gingivalis) fimbrillin (FimA) on the progression of esophageal squamous cell carcinoma (ESCC). Methods:Wild-type P. gingivalis and fimA gene-deleted P. gingivalis ( fimA-/-P. gingivalis) were used to infect ESCC cells after morphology and PCR identification. Immunofluorescence, CCK-8 and Transwell chamber were used to detect the effects of FimA on the infectivity of P. gingivalis and it influences on cell invasion, proliferation and migration. Western blot was used to detect pSmad2/3 changes. The growth of tumor was detected in a nude mouse model bearing subcutaneous tumor. Results:Deletion of FimA might reduce the interbacterial adhesion of P. gingivalis. Compared with wild-type P. gingivalis, less fimA-/-P. gingivalis could infect NE6-T cells. Moreover, the proliferation, migration and invasion of NE6-T and KYSE30 cells as well as the activation of pSmad2/3 induced by P. gingivalis were inhibited after deletion of FimA. The growth of KYSE30 infected by fimA-/-P. gingivalis in nude mice was significantly slower than that of the wild-type P. gingivalis group. Conclusions:FimA mediated the effects of P. gingivalis on promoting the evolution of ESCC and was a potential target molecule to block the tumor-promoting effect of P. gingivalis.

Objective: To compare the clinical efficacy of posterior atlantoaxial screw-rod fixation combined with spinous process muscle-vessel complex bone graft or iliac bone graft for atlantoaxial instability. Methods: A retrospective case-control study was conducted to analyze the clinical data of 56 patients with atlantoaxial instability admitted to the Sixth Hospital of Ningbo from September 2014 to October 2016. There were 35 males and 21 females, with the age range from 9 to 59 years [(50.3±3.2)years]. A total of 26 patients were treated with posterior atlantoaxial screw-rod fixation combined with spinous process muscle-vessel complex bone graft (complex group), while 30 patients were treated with iliac bone graft (ilium group). Patients showed different degrees of neck pain and limited neck activity preoperatively. X-ray films and three-dimensional CT examination of the cervical spine were taken before and after operation for evaluating the atlantoaxial reduction, bone graft fusion and internal fixation. The operation time, intraoperative bleeding, bone fusion time, visual analogue scale (VAS), Japanese Orthopaedic Association (JOA) score, atlantodental interval (ADI) and axial symptoms were compared between the two groups, and the complications were recorded. Results: Both groups were followed up for 24-30 months, with an average of 27.4 months. In the complex group and the ilium group, the operation time was (2.21±0.25)hours and (2.72±0.26)hours (P<0.01); the intraoperative blood loss was (227.3±45.4)ml and (277.7±43.4)ml, respectively (P<0.05); the bone fusion time was (6.9±0.5)months and (8.1±1.8)months (P>0.05), respectively. In the complex group, the VAS was (5.45±0.69)points before operation, (2.64±0.51)points at the follow-up one month after operation, (0.91±0.7)points at the follow-up 12 months after operation, and (0.45±0.16)points at the follow-up 24 months after operation; and in the ilium group, the VAS was (5.18±0.75)points, (2.45±0.52)points, (1.27±0.19)points and (0.41±0.18)points correspondingly. In terms of VAS, there were significant differences before and after operation within each group (P<0.01), while there were no significant differences between the two groups at different time points (P>0.05). In the complex group, the JOA score was (10.82±0.35)points before operation, (12.73±0.65)points at the follow-up one month after operation, (15.18±0.61)points at the follow-up 12 months after operation, and (15.64±0.15)points at the follow-up 24 months after operation; and in the ilium group, the JOA score was (10.73±1.19)points, (13.01±0.63)points, (14.73±0.91)points and (15.55±0.51)points correspondingly. In terms of JOA score, there were significant differences between before and after operation within each group (P<0.01), while there were no significant differences between the two groups at different time points (P>0.05). In the complex group, the ADI was (2.28±0.59)mm before operation, (1.83±0.56)mm at the follow-up one month after operation, (1.71±0.56)mm at the follow-up 12 months after operation, and (1.59±0.67)mm at the follow-up 24 months after operation; and in the ilium group, the ADI was (2.23±0.60)mm, (1.80±0.18)mm, (1.67±0.69)mm and (1.62±0.53)mm correspondingly. In terms of ADI, there were significant differences between before and after operation within each group (P<0.01), while there were no significant differences between the two groups at different time points (P>0.05). The axial symptom scores were graded as excellent in 23 patients and good in three patients of the complex group while excellent in 21 patients and good in nine patients in the ilium group (P>0.05). There were no patients with spinal nerve injury caused by pedicle screw placement after operation. One patient in the ilium group had incision errhysis and recovered after dressing change, and other patients had no incision infection. Conclusions For atlantoaxial instability, posterior atlantoaxial screw-rod fixation combined with spinous process muscle-vessel complex bone graft or autogenous iliac bone graft can both achieve satisfactory clinical results. The spinous process-muscle-vascellum complex graft has less operation time and intraoperative bleeding than the autogenous iliac bone graft, which can be a feasible alternative operation.

Objective:To investigate the role of an inflammatory microenvironment induced by Porphyromonasgingivalis ( P. gingivalis) in the occurrence of esophageal squamous cell carcinoma (ESCC) in mice. Methods:A total of 180 C57BL/6 mice were randomly divided into 6 groups, i.e. control group, P. gingivalis group, 4NQO group, 4NQO + P. gingivalis group, 4NQO + P. gingivalis + celecoxib group, and 4NQO + P. gingivalis + antibiotic cocktail (ABC, including metronidazole, neomycin, ampicillin, and vancomycin) group, with 30 mice in each group, using the random number table. All mice were normalized by treatment with ABC in drinking water for 2 weeks. In the following 2 weeks, the mice in the control group and the P. gingivalis group were given drinking water, while the other 4 groups were treated with 30 μg/ml 4NQO in the drinking water. In weeks 11-12, the mice in the P. gingivalis group, the 4NQO + P. gingivalis group, the 4NQO + P. gingivalis + celecoxib group, and the 4NQO + P. gingivalis + ABC group were subjected to ligation of the second molar in oral cavity followed by oral P. gingivalis infection thrice weekly for 24 weeks in weeks 11-34. In weeks 13-34, the mice in 4NQO + P. gingivalis+celecoxib group and 4NQO + P. gingivalis + ABC group were administered with celecoxib and ABC for 22 weeks, respectively. At the end of 34 weeks, gross and microscopic alterations were examined followed by RT-qPCR and immunohistochemistry to examine the expression profiles of inflammatory- and tumor-molecules in esophagi of mice. Results:At 34 weeks, 4NQO treatment alone did not affect the foci of papillary hyperproliferation, diseased area, and the thickness of the esophageal wall, but significantly enhanced the foci of hyperproliferation (median 1.00, P＜0.05) and mild/moderate dysplasia (median 2.00, P＜0.01). In addition, the expression levels of IL-6 [8.35(3.45,8.99)], IL-1β [6.90(2.01,9.72)], TNF-α [12.04(3.31,14.08)], c-myc [2.21(1.80,3.04)], pSTAT3, Ki-67, and pH2AX were higher than those in the control group. The pathological changes of the esophageal mucosa were significantly more overt in the 4NQO + P. gingivalis group in terms of the foci of papillary hyperproliferation (median 2.00), diseased area (median 2.51 mm 2), the thickness of the esophageal wall (median 172.52 μm), the foci of hyperproliferation (median 1.00, P＜0.05), and mild/moderate dysplasia (median 1.00, P＜0.01). In mice of the 4NQO + P. gingivalis group, the expression levels of IL-6 [12.27(5.35,22.08)], IL-1β [13.89(10.04,15.96)], TNF-α [19.56(6.07,20.36)], IFN-γ [11.37(8.23,20.07)], c-myc [2.62(1.51,4.25)], cyclin D1 [4.52(2.68,7.83)], nuclear pSTAT3, COX-2, Ki-67, and pH2AX were significantly increased compared with the mice in the control group. In mice of the 4NQO + P. gingivalis group, the diseased area, invasive malignant foci as well as pSTAT3 and pH2AX expression were significantly blunted by celecoxib. Treatment with ABC markedly reduced the papillary hyperproliferative foci, invasive malignant foci, and pSTAT3 expression but not pH2AX. Conclusions:P. gingivalis promotes the occurrence of esophageal squamous cell carcinoma in mice by inducing an inflammatory microenvironment primed with 4NQO induced DNA damage. Clearance of P. gingivalis with ABC or anti-inflammatory intervention holds promise for prevention of esophageal squamous cell malignant pathogenesis via blockage of IL-6/STAT3 signaling and amelioration of inflammation.

Objective:To investigate the role of an inflammatory microenvironment induced by Porphyromonasgingivalis ( P. gingivalis) in the occurrence of esophageal squamous cell carcinoma (ESCC) in mice. Methods:A total of 180 C57BL/6 mice were randomly divided into 6 groups, i.e. control group, P. gingivalis group, 4NQO group, 4NQO + P. gingivalis group, 4NQO + P. gingivalis + celecoxib group, and 4NQO + P. gingivalis + antibiotic cocktail (ABC, including metronidazole, neomycin, ampicillin, and vancomycin) group, with 30 mice in each group, using the random number table. All mice were normalized by treatment with ABC in drinking water for 2 weeks. In the following 2 weeks, the mice in the control group and the P. gingivalis group were given drinking water, while the other 4 groups were treated with 30 μg/ml 4NQO in the drinking water. In weeks 11-12, the mice in the P. gingivalis group, the 4NQO + P. gingivalis group, the 4NQO + P. gingivalis + celecoxib group, and the 4NQO + P. gingivalis + ABC group were subjected to ligation of the second molar in oral cavity followed by oral P. gingivalis infection thrice weekly for 24 weeks in weeks 11-34. In weeks 13-34, the mice in 4NQO + P. gingivalis+celecoxib group and 4NQO + P. gingivalis + ABC group were administered with celecoxib and ABC for 22 weeks, respectively. At the end of 34 weeks, gross and microscopic alterations were examined followed by RT-qPCR and immunohistochemistry to examine the expression profiles of inflammatory- and tumor-molecules in esophagi of mice. Results:At 34 weeks, 4NQO treatment alone did not affect the foci of papillary hyperproliferation, diseased area, and the thickness of the esophageal wall, but significantly enhanced the foci of hyperproliferation (median 1.00, P＜0.05) and mild/moderate dysplasia (median 2.00, P＜0.01). In addition, the expression levels of IL-6 [8.35(3.45,8.99)], IL-1β [6.90(2.01,9.72)], TNF-α [12.04(3.31,14.08)], c-myc [2.21(1.80,3.04)], pSTAT3, Ki-67, and pH2AX were higher than those in the control group. The pathological changes of the esophageal mucosa were significantly more overt in the 4NQO + P. gingivalis group in terms of the foci of papillary hyperproliferation (median 2.00), diseased area (median 2.51 mm 2), the thickness of the esophageal wall (median 172.52 μm), the foci of hyperproliferation (median 1.00, P＜0.05), and mild/moderate dysplasia (median 1.00, P＜0.01). In mice of the 4NQO + P. gingivalis group, the expression levels of IL-6 [12.27(5.35,22.08)], IL-1β [13.89(10.04,15.96)], TNF-α [19.56(6.07,20.36)], IFN-γ [11.37(8.23,20.07)], c-myc [2.62(1.51,4.25)], cyclin D1 [4.52(2.68,7.83)], nuclear pSTAT3, COX-2, Ki-67, and pH2AX were significantly increased compared with the mice in the control group. In mice of the 4NQO + P. gingivalis group, the diseased area, invasive malignant foci as well as pSTAT3 and pH2AX expression were significantly blunted by celecoxib. Treatment with ABC markedly reduced the papillary hyperproliferative foci, invasive malignant foci, and pSTAT3 expression but not pH2AX. Conclusions:P. gingivalis promotes the occurrence of esophageal squamous cell carcinoma in mice by inducing an inflammatory microenvironment primed with 4NQO induced DNA damage. Clearance of P. gingivalis with ABC or anti-inflammatory intervention holds promise for prevention of esophageal squamous cell malignant pathogenesis via blockage of IL-6/STAT3 signaling and amelioration of inflammation.