Objective:To investigate spinal cord morphological changes and risk factors in upper cervical spine surgery using C 2 medial "in-out-in" pedicle screws in patients with atlantoaxial dislocation (AAD) and high-riding vertebral artery (HRVA). Methods:A retrospective analysis was conducted on 41 patients with AAD who underwent C 2 medial "in-out-in" pedicle screw implantation at our hospital between January 2019 and December 2023. The cohort included 12 males and 29 females, with a mean age of 47.6±10.3 years (range: 18-68 years). Among them, 30 patients received unilateral C 2 medial "in-out-in" screws, while 11 patients received bilateral screws. All patients underwent posterior reduction and internal fixation. Measurements included C 2 pedicle height, C 2 pedicle width, C 2 horizontal spinal canal width, screw invasion distance into the spinal canal, and spinal canal invasion rate based on CT findings. MRI evaluations included spinal canal-dura mater distance, dura mater-spinal cord distance, spinal canal-spinal cord distance, and spinal cord cross-sectional area. The change rate of spinal cord cross-sectional area was calculated and categorized into >20% decrease group (decreasing group) and ≤20% decrease group (non-decreasing group). Postoperative outcomes were assessed using the Visual Analogue Scale (VAS) and Japanese Orthopedic Association (JOA) scores. Results:The mean operative time was 165.5±30.1 minutes (range: 120-220 minutes). No spinal cord vascular injuries or severe complications were observed. However, five patients experienced cerebrospinal fluid (CSF) leakage, requiring delayed removal of drainage tubes, but their wounds healed successfully. Follow-ups (range: 6-24 months) showed no loosening of internal fixation, fractures, or bone graft nonunion, with a 100% bone fusion rate at 6 months postoperatively. VAS scores improved significantly from a preoperative median of 5.0 (3.5, 6.0) (range: 1-7) to 2.0 (2.0, 3.0) at 3 months and 1.0 (1.0, 1.0) at 6 months ( Z=36.716, P< 0.001). JOA scores improved from 10.0 (9.0, 12.0) (range: 4-14) preoperatively to 13.0 (11.0, 13.5) at 3 months and 14.0 (12.0, 15.0) at 6 months ( Z=67.093, P<0.001). The height of C 2 pedicle was 5.50±1.78 mm, the width of C 2 pedicle was 2.27±1.23 mm, the width of C 2 horizontal spinal canal was 23.76±4.91 mm, the spinal canal-dura mater distance was 3.08±0.85 mm, dura mater-spinal cord distance was 3.23±0.85 mm, the spinal canal-spinal cord distance was 6.31±1.11 mm, the distance of screw invasion into the spinal canal was 2.80±1.54 mm, the rate of spinal canal invasion was 12.1%±6.8%, the preoperative spinal cord cross-sectional area was 69.81±13.27 mm 2, and the postoperative spinal cord cross-sectional area was 68.81±13.94 mm 2. Based on spinal cord cross-sectional area changes, 32 patients were classified into the non-decreasing group, and 9 patients into the decreasing group. The intraclass correlation coefficient (ICC) values ranged from 0.733 to 0.984, indicating high measurement reliability. There were significant differences in the proportion of C 2 bilateral medial "in-out-in" screws (χ 2=6.903, P=0.009), the incidence of CSF leakage (χ 2=15.391, P<0.001), the distance of screw invasion into the spinal canal ( t=4.990, P<0.001) and the rate of spinal canal invasion ( t=4.045, P<0.001) in the decreasing group versus the non-decreasing group. The JOA scores of the non-decreasing group were significantly higher at 3 and 6 months postoperatively compared to the decreasing group ( P<0.05). No other parameters showed significant differences between the groups. Binary logistic regression analysis identified spinal canal invasion rate [ OR=1.963, 95% CI (1.010, 3.817), P=0.047] as an independent risk factor for spinal cord cross-sectional area reduction. The Jordan index was 0.875, with a spinal canal invasion rate threshold of 14.18%, a sensitivity of 1.000, a specificity of 0.875, and an AUC of 0.983, indicating strong predictive value. Conclusion:The C 2 medial "in-out-in" screw technique provides effective posterior fixation and fusion for AAD patients with HRVA. However, to minimize spinal cord morphological changes and associated risks, the spinal canal invasion rate should be kept below 14.18% when using this technique.

Objective:To investigate spinal cord morphological changes and risk factors in upper cervical spine surgery using C 2 medial "in-out-in" pedicle screws in patients with atlantoaxial dislocation (AAD) and high-riding vertebral artery (HRVA). Methods:A retrospective analysis was conducted on 41 patients with AAD who underwent C 2 medial "in-out-in" pedicle screw implantation at our hospital between January 2019 and December 2023. The cohort included 12 males and 29 females, with a mean age of 47.6±10.3 years (range: 18-68 years). Among them, 30 patients received unilateral C 2 medial "in-out-in" screws, while 11 patients received bilateral screws. All patients underwent posterior reduction and internal fixation. Measurements included C 2 pedicle height, C 2 pedicle width, C 2 horizontal spinal canal width, screw invasion distance into the spinal canal, and spinal canal invasion rate based on CT findings. MRI evaluations included spinal canal-dura mater distance, dura mater-spinal cord distance, spinal canal-spinal cord distance, and spinal cord cross-sectional area. The change rate of spinal cord cross-sectional area was calculated and categorized into >20% decrease group (decreasing group) and ≤20% decrease group (non-decreasing group). Postoperative outcomes were assessed using the Visual Analogue Scale (VAS) and Japanese Orthopedic Association (JOA) scores. Results:The mean operative time was 165.5±30.1 minutes (range: 120-220 minutes). No spinal cord vascular injuries or severe complications were observed. However, five patients experienced cerebrospinal fluid (CSF) leakage, requiring delayed removal of drainage tubes, but their wounds healed successfully. Follow-ups (range: 6-24 months) showed no loosening of internal fixation, fractures, or bone graft nonunion, with a 100% bone fusion rate at 6 months postoperatively. VAS scores improved significantly from a preoperative median of 5.0 (3.5, 6.0) (range: 1-7) to 2.0 (2.0, 3.0) at 3 months and 1.0 (1.0, 1.0) at 6 months ( Z=36.716, P< 0.001). JOA scores improved from 10.0 (9.0, 12.0) (range: 4-14) preoperatively to 13.0 (11.0, 13.5) at 3 months and 14.0 (12.0, 15.0) at 6 months ( Z=67.093, P<0.001). The height of C 2 pedicle was 5.50±1.78 mm, the width of C 2 pedicle was 2.27±1.23 mm, the width of C 2 horizontal spinal canal was 23.76±4.91 mm, the spinal canal-dura mater distance was 3.08±0.85 mm, dura mater-spinal cord distance was 3.23±0.85 mm, the spinal canal-spinal cord distance was 6.31±1.11 mm, the distance of screw invasion into the spinal canal was 2.80±1.54 mm, the rate of spinal canal invasion was 12.1%±6.8%, the preoperative spinal cord cross-sectional area was 69.81±13.27 mm 2, and the postoperative spinal cord cross-sectional area was 68.81±13.94 mm 2. Based on spinal cord cross-sectional area changes, 32 patients were classified into the non-decreasing group, and 9 patients into the decreasing group. The intraclass correlation coefficient (ICC) values ranged from 0.733 to 0.984, indicating high measurement reliability. There were significant differences in the proportion of C 2 bilateral medial "in-out-in" screws (χ 2=6.903, P=0.009), the incidence of CSF leakage (χ 2=15.391, P<0.001), the distance of screw invasion into the spinal canal ( t=4.990, P<0.001) and the rate of spinal canal invasion ( t=4.045, P<0.001) in the decreasing group versus the non-decreasing group. The JOA scores of the non-decreasing group were significantly higher at 3 and 6 months postoperatively compared to the decreasing group ( P<0.05). No other parameters showed significant differences between the groups. Binary logistic regression analysis identified spinal canal invasion rate [ OR=1.963, 95% CI (1.010, 3.817), P=0.047] as an independent risk factor for spinal cord cross-sectional area reduction. The Jordan index was 0.875, with a spinal canal invasion rate threshold of 14.18%, a sensitivity of 1.000, a specificity of 0.875, and an AUC of 0.983, indicating strong predictive value. Conclusion:The C 2 medial "in-out-in" screw technique provides effective posterior fixation and fusion for AAD patients with HRVA. However, to minimize spinal cord morphological changes and associated risks, the spinal canal invasion rate should be kept below 14.18% when using this technique.

Objective:To analyze the clinical efficacy of three-dimensional(3D) reconstruction guided uniportal fluorescence thoracoscopic subsegmentectomy for the pulmonary nodules.Methods:We retrospectively analyzed 50 patients with nodules who underwent uniportal fluorescence thoracoscopic subsegmentectomy from December 2021 to February 2024. All patients underwent thin-slice CT scanning and 3D reconstruction preoperatively. 12 patients were given CT-guided hookwire localization preoperatively.The intersegmental plane was identified by fluorescence method.Results:One patient was converted to right upper lobectomy due to no lesion found in S1b. The mean blood loss was(23.4±16.5)ml and the mean operative time was(126.5±38.5)min. The mean duration of postoperative drainage was(2.6±0.8)days. Mean postoperative hospitalization was(4.8±1.8)days. There were 2 cases with postoperative pulmonary infections, including one with encapsulated pleural effusion. There was no air leakage over 3 days, and no death within 30 days after surgery.Conclusion:3D reconstruction guided uniportal fluorescence thoracoscopic subsegmentectomy is a safe and feasible technique for resection of pulmonary nodules in lung subsegments, and surgical indications must be strictly controlled.

Objective    To summarize the clinical experience of thoracoscopic combined subsegmentectomy (CSS). Methods    The clinical data of 76 patients who underwent thoracoscopic CSS in Anqing Municipal Hospital from May 2018 to July 2022 were retrospectively analyzed, including 22 males and 54 females, aged 27.0-76.0 (54.3±10.5) years. All patients underwent preoperative three-dimensional computed tomography bronchography and angiography using dual source CT. The modified inflation-deflation technique or indocyanine green was used to identify the intersubsegmental border. Results    A total of 86 pulmonary nodules were resected in 76 patients. One patient of left upper lobe S1+2c+S4a, 1 patient of right upper lobe S2b+S3a and 1 patient of right upper lobe S1b+S3b were further performed lobectomy due to insufficient margin. One patient of left upper lobe S1+2+S3a was further performed left upper division segmentectomy due to residual atelectasis. One patient of left upper lobe S1+2c+S3a was further performed left upper division segmentectomy due to B3b+c injury, and the rest completed planned surgeries successfully. The operative time was 90.0-350.0 (174.9±53.2) min. The operative hemorrhage volume was 50.0 (20.0, 50.0) mL. The postoperative hospital stay time was 6.0 (5.0, 7.0) d. Postoperative complications included pulmonary infection in 9 patients, hemoptysis in 3 patients, persistent pulmonary leakage>3 d in 4 patients, pneumothorax in 1 patient, pleural effusion in 1 patient, and myocardial infarction in 1 patient. All of the patients were cured and discharged without perioperative death. Conclusion    Thoracoscopic CSS is relatively complex. Preoperative planning under three-dimensional reconstruction and intraoperative fine operation are helpful for safe completion.

Objective To explore the immunopathological mechanism for the imbalance between the positive signal mediated by inducible costimulator (ICOS) and the negative signal mediated by programmed death-1 (PD-1) in patients with myasthenia gravis (MG).Methods Eighty-two patients with MG,56 healthy controls (HC) and 20 non-MG (NMG) patients,collected in the First Affiliated Hospital of Suzhou University from February 2014 to December 2016,were chosen to participate in the study.The expression of ICOS and PD-1 on peripheral blood mononuclear cells was detected by immuno-fluorescence staining and flow cytometry.The levels of soluble programmed death-1 (sPD-1),soluble programmed death ligand 1 (sPD-L1),IL-4 and other cytokines were detected by enzyme-linked immunosorbent assay.Results (1) Flow cytometry analysis:The co-expression of PD-1,ICOS on CD4 + T cells from MG group (9.64％ (8.82％)) was higher than in HC (1.81％ (2.10％),Z =-7.389,P ＜0.05) and NMG group (2.86％ (1.49％),Z =-4.636,P ＜ 0.05).The expression of ICOS on CD4 + T cells,ICOS ligand (ICOSL) on CD14+ monocytes and CD19+ B cells were increased in MG group comparing with that of the control groups.The proportion of PD-1 + CD4 + T cells (MG group 16.82％ (10.66％),HC 9.34％ (9.18％),Z =-4.345,P＜0.05;NMG group 7.07％ (3.40％),Z=-4.594,P＜0.05) and PD-1 Ligand (PD-L1) + CD14+ monocytes was higher in MG patients.All of these were detected by flow cytometry.(2) ELISA analysis:Serum sPD-1 expression significantly increased in MG group compared with that in the control groups (MG group (1.87 ± 0.64) ng/ml,NMG group (1.49 ± 0.70) ng/ml,t =2.04,P ＜ 0.05;HC (1.05 ± 0.50)ng/ml,t =2.08,P ＜ 0.05),while for serum sPD-L1,there was no significant difference between MG and control groups.(3) Serum cytokines detection:The expression of IL-4 was increased in MG patients (MG group (61.88 ±5.15) pg/ml,HC (32.03 ±1.84) pg/ml,t=2.50,P＜0.05;NMG group (42.62± 3.31) pg/ml,t =2.34,P ＜0.05),and there was a negative correlation between the expression of sPD-1 and the concentration of IL-4.Conclusions The increased expression of PD-1 + ICOS + CD4 + T cells suggested the subset involved in the pathological progress of MG.sPD-1 might disturb the ligation of PD-1 on T cells and PD-L1 on antigen presenting cells,while the ligation of ICOS and ICOSL passed positive signal,leading to over activity of the subsets and the progression of disease.

Objective To explore the mechanism of sulfotanshinone sodium injection in treatment of patients with sudden deafness (SD).Methods Sixty patients with SD admitted to the Department of Otorhinolaryngology of Wuxi Traditional Chinese and Western Medicine Hospital from January to December 2016 were enrolled, and they were randomly divided into a study group and a control group (each 30 cases). The same basic treatment was given in the two groups, the patients in the study group were treated with sulfotanshinone sodium 40 mg intravenous (IV) drip, while the patients in the control group were treated with vinpocetine sodium chloride 20 mg IV drip, once a day for consecutive 14 days to complete a therapeutic course, and two courses were carried out in bothgroups. Before and after treatment, the changes of hearing threshold, indexes of hemorheology and immune function were compared between the patients in the two groups, and the clinical efficacy and adverse reactions in the two groups were observed.Results After treatment, the hearing threshold, hemorheology indexes, immune function index of CD8+ were significantly lower than those before treatment, while the CD3+, CD4+, CD4+/CD8+ ratio were significantly higher than those before treatment in the two groups, and the above changes of indexes were more obvious in the study group than those in the control group hearing [threshold (dB): 16.63±2.04 vs. 17.15±1.88, plasma viscosity (PV, mPa·s): 1.27±0.14 vs. 1.31±0.11, whole blood middle shearing viscosity (mPa·s): 4.77±0.33 vs. 4.95±0.28, whole blood high shearing viscosity (mPa·s): 3.86±0.25 vs. 4.00±0.31, erythrocyte aggregation index (EAI): 1.57±0.29 vs. 1.72±0.34, CD3+: 0.70±0.05 vs. 0.64±0.05, CD4+: 0.43±0.04 vs 0.37±0.03, CD8+: 0.32±0.04 vs. 0.34±0.03, CD4+/CD8+: 1.36±0.32 vs. 1.18±0.27]; the degree of whole blood low shearing viscosity (mPa·s: 6.72±0.80 vs. 7.01±1.13) and hematocrit (HCT: 0.38±0.04 vs. 0.40±0.03) decreasing weremore significant in the control group than those in the study group. The total effective rate was higher in study group than that in the control group [86.67% (26/30) vs. 83.33% (25/30)], but the difference between the two groups was not statistically significant (P > 0.05); the incidence of adverse reactions in the study group was markedly lower than that in the control group [3.33% (1/30) vs. 20.00% (6/30),P < 0.05].Conclusions Sulfot anshinone sodium injection can effectively enhance the SD patients' hearing, and improve their hemorheology indexes and immune function; the therapeutic results of sulfotanshinone sodium injection in safety and improvement in immune function are superior to those of vinpocetine sodium chloride injection.

Objective To discuss the clinical significance of programmed cell death-1 (PD-1) in neuromyelitis optica spectrum disorder (NMOSD) patients by analyzing PD-1 and programmed death 1-1igand (PD-L1) expressions.Methods Sixteen patients with NMOSD,16 patients with longitudinally extensive transverse myelitis (LETM),13 patients with opticneuritis (ON),20 with other diseases of the central nervous system (OTH) and 16 health controls (CONs) were chosen in our hospital from April 2015 to July 2016;their peripheral blood was separately collected.The PD-1 expression in the CD4+r lymphocytes,and PD-L1 expressions in the CD14+ mononuclear leucocytes and CD19+B lymphocytes of peripheral blood were detected by flow cytometry.ELISA was performed to analyze the levels of soluble PD-1 and soluble PD-L1 in plasma samples.Results The PD-1 level from the peripheral blood of NMOSD patients was significantly higher than that from LETM,ON,and OTH patients and CONs (P＜0.05).The PD-L1 level of NMOSD patients was significantly higher than that of the other 4 groups (P＜0.05).ELISA indicated that levels of soluble PD-1 and soluble PD-L1 in plasma samples from NMOSD patients were significantly higher than those in LETM,ON,and OTH patients and CONs (P＜0.05).Conclusion The PD-1/PD-L1 pathway is an important immune response approach and takes part in the earlier stage of the NMOSD pathological process.

Objective To investigate the feasibility of using leukocytes that were filtered out by LeukoReduction System ( LRS) to replace conventional human peripheral blood leukocytes in experimental researches and to comparatively analyze the differences between them in vitro biological functions and pheno-types of T cells. Methods Mononuclear cells were isolated from LRS-separated leukocytes and whole blood sample that collected from the same person by using Ficoll. Fluorescence-activated cell sorting ( FACS) was performed to analyze the phenotypes of T cells. CD3+T cells were sorted out by using magnetic beads. The T cells that were collected by using two different ways were incubated with anti-CD3 and anti-CD28 antibodies and IL-2 in vitro for 10 days. Several assays including cell counting, FACS and cytometric beads array ( CBA) were performed to comparatively analyze the differences in biological functions and phenotypes of T cells that were isolated by different methods. Results The phenotypes of T cells isolated from LRS filter and whole blood sample were highly similar at the initial stage. The sorting rate of CD3+T cells form LRS filter reached a high level and met the requirements for experimental researches. No statistically significant differ-ences in cell count, phenotype, expression of costimulatory molecules and cytokine secretion were observed between T cells isolated from LRS filter and whole blood sample. Conclusion This study suggested that the T cells isolated from LRS filter could be used as an alternative to whole blood T cells for fundamental resear-ches since they were similar in cell vitality, phenotype and biological functions. It provided a new way to solve the problem of blood shortage in clinic and scientific research.

Objective To investigate the role and mechanism of B7-H4, a negative costimulatory molecule, in mediating the immunomodulatory effects of mesenchymal stem cells C3H10T1/2 (C3H10) on T cell polarization. Methods The lentiviral vectors that carried the shRNA targeting mouse B7-H4 were transfected into mouse mesenchymal stem cells (C3H10-B7-H4). The cells were co-cultured with PHA-acti-vated mice spleen lymphocytes before and after the transfection. ELISA was performed to detect the concen-trations of cytokines in supernatants of cell culture in order to elucidate the effects of B7-H4 expressed by C3H10 on T cell polarization. A mouse model of experimental allergic encephalitis (EAE) was established. Fifty C57BL/6 mice were divided into five groups including control group, EAE group, C3H10 group (injec-ting EAE mice with C3H10 cells), C3H10-NC group ( injecting EAE mice with C3H10-NC cells) and C3H10-B7-H4 group (injecting EAE mice with C3H10-B7-H4 cells). ELISA was performed to detect the soluble form of IL-2, IL-17, IFN-γ and IL-4 in plasma samples. Results Knocking down the B7-H4 gene with shRNA significantly decreased the expression of B7-H4 on C3H10 cells, which weakened the inhibitory effects of C3H10 cells on the secretion of IL-2, IL-17 and IFN-γ by spleen lymphocytes. The therapeutic effects of C3H10-B7-H4 cells on mice with EAE were weakened after silencing the B7-H4 gene expression, which was manifested as higher nerve function score and earlier onset and bring forwarded peak time of EAE than those of the C3H10 group. Treating EAE mice with C3H10-B7-H4 cells was less efficient in inhibiting the expression of IL-2, IL-17 and IFN-γin plasma. However, knocking down the B7-H4 gene had no signif-icant effect on the expression of IL-4 in terms of treating EAE with C3H10 cells. Conclusion The co-inhib-itor molecule B7-H4 expressed on C3H10 cells mediated the treatment of EAE with C3H10 cells by regula-ting Th1 and Th17 effector T cells.

Objective:To explore a new method of the cultivation of adoptive immunotherapy cells.Methods: Mononuclear cells was isoplated by density gradient centrifugation and then proliferated by using CD 40-agonist monoclonal antibody 5C11、cytokine of IFN-αand IL-7(CD40 group)in vitro.During the culturing procedure ,the cell morphology was obersved by optical microscope.The percentage of T-lymphocytes, NK-T cells, Treg cells and the cell proliferation, which were compared with CIK group CD3mAb activated,was detected on the 9th day.Results:There was no significant difference of CD 4+/CD8+T cells percentage between the two groups.But the Treg cells percentage of CIK group was far higher than that of CD 40 group,while the percentage of CD3+CD56+NK-T cells was lower than that of CD 40 group.And a group of Mo-NK-DC cells were observed in the CD 40 group.Conclusion: The new method of adoptive immunity therapy has been established in this study could increase the percentage of NK -T cells which had the ability to kill tumor cells.Simultaneously ,it is reduced the amount of Treg cells significantly.