1.Posterior lateral perforator flap in lower limb combined with free fibula for maxillary tissue defect repair.
Mingming YAN ; Luwen SONG ; Zhenghao MA ; Tao WANG ; Kai HU ; Xuji WANG ; Jiancheng LI
Chinese Journal of Reparative and Reconstructive Surgery 2025;39(1):88-94
OBJECTIVE:
To investigate the effectiveness of posterior lateral perforator flap in lower limb combined with free fibula for maxillary tissue defect repair.
METHODS:
Between December 2018 and December 2023, 16 patients with the maxillary malignant tumors were admitted. There were 10 males and 6 females, with an average age of 64.3 years (range, 54-75 years). There were 7 cases of maxillary gingival cancer, 5 cases of hard palate cancer, and 4 cases of maxillary sinus cancer. According to the 2017 American Joint Committee on Cancer (AJCC) TNM stage, there were 8 cases of stage Ⅲ, 6 cases of stage Ⅳa, and 2 cases of stage Ⅳb. After resection of the lesion, the remaining maxillary defects were classified into class Ⅱa in 3 cases, class Ⅱb in 5 cases, and class Ⅲb in 8 cases according to Brown's classification. The size of soft tissue defects ranged from 4 cm×3 cm to 8 cm×6 cm. The posterior lateral perforator flap in lower limb in size of 5 cm×4 cm-9 cm×7 cm were harvested to repair soft tissue defects, and free fibula in length of 6-11 cm were used to repair bone defects. The donor sites of the lower limb were sutured directly (6 cases) or repaired with free skin grafting (10 cases). Six patients with positive lymph node pathology were treated with radiotherapy after operation. At 6 and 12 months after operation, the self-assessment was performed by the University of Washington Quality of Survival Questionnaire Form (QUW-4) in five dimensions (facial appearance, swallowing function, chewing function, speech function, and mouth opening), and swallowing function was evaluated by using the Kubota water swallowing test.
RESULTS:
Postoperative pathological examination showed that all patients were squamous cell carcinoma. One patient who was treated with radiotherapy developed osteomyelitis and 1 patient developed venous crisis of skin flap. The rest of the flaps and all skin grafts survived, and the wounds healed by first intention. All patients were followed up 1-5 years (mean, 2.8 years). Two patients died of local recurrence of the tumor at the 4th and 5th years after operation, respectively. Except for the chewing function score and total score at 6 months after operation, which showed significant differences compared to preoperative scores ( P<0.05), there was no significant difference in other QUW-4 scale scores between different time points ( P>0.05). The patients' swallowing function evaluated by Kubota water swallowing test reached normal in 4 cases, suspicious in 9 cases, and abnormal in 3 cases at 6 months after operation, and 10, 6, and 0 cases at 12 months after operation, respectively. The swallowing function at 12 months was significantly better than that at 6 months ( Z=-2.382, P=0.017).
CONCLUSION
The posterior lateral perforator flap in the lower limb combined with free fibula to repair maxillary tissue defects can repair soft and hard tissue defects at the same time, so that the patient's facial appearance, swallowing function, chewing function, speech function, and mouth opening are satisfactorily restored and the mid-term effectiveness is good.
Humans
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Middle Aged
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Male
;
Female
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Fibula/surgery*
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Aged
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Perforator Flap
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Plastic Surgery Procedures/methods*
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Maxilla/surgery*
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Maxillary Neoplasms/surgery*
;
Free Tissue Flaps/transplantation*
;
Lower Extremity/surgery*
;
Bone Transplantation/methods*
;
Treatment Outcome
2.Effect and mechanism of LncRNA EFRL on homocysteine-induced atherosclerosis in macrophage efferocytosis.
Jiaqi YANG ; Zhenghao ZHANG ; Fang MA ; Tongtong XIA ; Honglin LIU ; Jiantuan XIONG ; Shengchao MA ; Yideng JIANG ; Yinju HAO
Chinese Journal of Cellular and Molecular Immunology 2025;41(7):577-584
Objective To investigate the effect and mechanism of Efferocytosis Relatived LncRNA (EFRL) on homocysteine-induced atherosclerosis in macrophage efferocytosis. Methods RAW264.7 cells were cultured in vitro, and the Control group (0 μmol/L Hcy) and Hcy intervention group (100 μmol/L Hcy) were set up. After GapmeR transfection of macrophages with Hcy intervention, EFRL knockdown negative control group (Hcy combined with LNA-NC) and EFRL knockdown group (Hcy combined with LNA-EFRL) were set up. High-throughput sequencing was applied for different expression of LncRNA MSTRG. 88917.16 (EFRL), UCSC was used to analyze its conservation, CPC and CPAT were used to analyze its ability to encode proteins, and GO and KEGG were used to analyze related biological functions. The localization of LncRNA EFRL in macrophages was analyzed by nucleoplasmic separation and RNA-FISH. Quantitative real-time PCR was used to detect the expression levels of LncRNA EFRL and its target gene SPAST in Hcy-treated macrophages. The apoptosis rate of Jurkat cells induced by UV was detected by flow cytometry. In vitro efferocytosis assay combined with immunofluorescence technique was used to analyze macrophage efferocytosis. ELISA was used to detect the levels of interleukin 1β(IL-1β) and IL-18. Results The new LncRNA MSTRG.88917.16 was identified and named EFRL(Efferocytosis Relatived LncRNA). UCSC, CPC and CPAT analyses showed that LncEFRL is highly conserved and does not have the ability to encode proteins. GO and KEGG analyses suggested that LncEFRL may be involved in macrophage efferocytosis. LncRNA EFRL was localized in the nucleus of macrophages as determined by nucleoplasmic separation and RNA-FISH. In comparison to the Control group, the expression levels of LncRNA EFRL and its target gene SPAST in the Hcy group were increased. In comparison to the Control group (0 min), the apoptosis rate of the experimental group (15, 30 min) Annexin V is more than 85%. Compared with Hcy combined with LNA-NC group, Hcy combined with LNA-EFRL group had enhanced macrophage efferocytosis and reduced levels of inflammatory factors. Compared with Hcy combined with LNA-NC group, the expression level of SPAST in Hcy combined with LNA-EFRL group was decreased. Conclusion Inhibition of EFRL expression can alleviate the process of Hcy inhibiting macrophage efferocytosis, and the mechanism is related to the regulation of the downstream target gene SPAST by EFRL.
RNA, Long Noncoding/physiology*
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Animals
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Homocysteine
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Mice
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Macrophages/drug effects*
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Humans
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RAW 264.7 Cells
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Atherosclerosis/chemically induced*
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Apoptosis/genetics*
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Phagocytosis/genetics*
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Jurkat Cells
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Interleukin-1beta/genetics*
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Efferocytosis
3.Clinical analysis of mandibular tumor resection with free fibula transplantation and implant implantation via the intraoral approach.
Jiancheng LI ; Mingming YAN ; Zhenghao MA ; Ruixue TIAN ; Xuji WANG ; Kai HU ; Lina JIANG
West China Journal of Stomatology 2025;43(2):212-219
OBJECTIVES:
To investigate the clinical application of the digital-assisted reconstruction of the mandible and tumors with free fibula transplantation and immediate implantation via the intraoral approach.
METHODS:
Twelve patients with benign mandibular tumors were collected. Three-dimensional mandibular reconstruction was performed digitally before surgery to simulate mandibular tumor resection, fibula resection and reconstruction, and implant implantation. The intraoperative resection of the mandibular tumor was conducted through the intraoral approach under the guidance of a guide plate, and fibula resection, molding, reconstruction, and oral fixation were immediately performed. Implant implantation was performed during the second phase of implant surgery and denture restoration was performed 1-2 months after surgery.
RESULTS:
The types of mandibular defects were BrownⅠ (one case), Ⅰc (four cases), Ⅱ (one case), Ⅱc(three cases), and Ⅲ (three cases). The length of the fibular bone was 12-22 cm. The number of fibular molding amputations was as follows: two cases in two segments, six cases in three segments, three cases in four segments, and one case in five segments. All of these cases underwent folding fibular reconstruction of mandibular and alveolar bone defects. A total of 44 implants were implanted, and none failed after operation.
CONCLUSIONS
The intraoral approach is a reliable method for the resection of mandibular benign tumors, with few postoperative complications and the ability to position and fix accurately the reconstructed folded fibula under digital design. The immediate implantation of the transplanted fibula does not affect the blood supply and has a high success rate. It is an effective and reliable method for the resection and reconstruction of mandibular benign tumors.
Humans
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Fibula/transplantation*
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Mandibular Neoplasms/surgery*
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Mandibular Reconstruction/methods*
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Bone Transplantation/methods*
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Male
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Middle Aged
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Female
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Mandible/surgery*
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Adult
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Free Tissue Flaps
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Surgery, Computer-Assisted
4.Effect of LncRNA SNHG1 on homocysteine-induced podocytespyrophosis
Zhenghao ZHANG ; Fang MA ; Qing ZHANG ; Tongtong XIA ; Honglin LIU ; Zhigang BAI ; Guanjun LU ; Jingwen ZHANG ; Hongjian PENG ; Yideng JIANG ; Shengchao MA
The Journal of Practical Medicine 2024;40(4):476-482
Objective To investigate the role of lncRNA SNHG1 in homocysteine-induced pyroptosis of podocyte.Methods Cbs+/-mice were randomly divided into two groups:a normal diet group(ND)and a high me-thionine diet group(HMD).Western blotting was used to detect the protein expression levels of Caspase-1,Cleaved Caspase-1,and NLRP3.Mouse renal glomerular podocytes were cultured in vitro,and then assigned into a control group(Control,0 μmol/L Hcy)and a homocysteine intervention group(Hcy,80 μmol/L Hcy).Western blotting was used to detect the protein expression levels of Caspase-1,Cleaved Caspase-1,and NLRP3.Mouse renal glomerular podocyion group(OE-NC + Hcy)and the lncSNHG1 overexpression + homocysteine intervention group(OE-SNHG1 + Hcy)were also established.After 48 hours of intervention,Real-time fluorescence quantita-tive PCR was used to detect the expression of lncSNHG1 in podocytes after Hcy intervention.Western blot was used to detect the expressions of Caspase-1,Cleaved Caspase-3 and NLRP3.Immunofluorescence was used to de-tect the expression levels of GSDMD and GSDMD-N.ELISA was used to detect the contents of IL-1β and IL-18.Results(1)In the animal experiments,the expression levels of pyroptosis-related proteins Caspase-1,Cleaved Caspase-1,NLRP3,GSDMD,and GSDMD-N were all increased in the HMD group compared with the ND group.(2)In the cellular experiments,the expression levels of Caspase-1,Cleaved Caspase-1,NLRP3,GSDMD,and GSDMD-N were all increased in the Hcy group compared with the Control group,and the contents of pyroptosis-mediated inflammatory factors IL-1β and IL-18 were increased as well.(3)In the cellular experiments,the expres-sion of lncSNHG1 was increased in the Hcy group compared with the control group.After transduction with lnc-SNHG1 lentivirus,the expression of lncSNHG1 was increased in the OE-SNHG1 group,compared with the control group and the OE-NC group.(4)In the cellular experiments,the expressions of pyroptosis-related proteins Cas-pase-1,Cleaved Caspase-1,NLRP3,GSDMD,and GSDMD-N were increased compared with the OE-NC+Hcy group,and the contents of pyroptosis-mediated inflammatory factors IL-1β and IL-18 were increased in the OE-SNHG1+Hcy group.Conclusion These results indicate that lncSNHG1 may play a role in promoting Hcy induced podocytepyroptosis.
5.Application value of artificial intelligence in surgical precision diagnosis and treatment of rectal cancer
Zhanwei FU ; Zhenghao CAI ; Junjun MA
Chinese Journal of Gastrointestinal Surgery 2024;27(6):574-578
Colorectal cancer is the most common malignant tumor of digestive tract, and the incidence of colorectal cancer in China is especially characterized by middle and low rectal cancer. In recent years, with the progress of computer science, artificial intelligence technology has developed rapidly, and has achieved a lot of application results in the medical field. At present, artificial intelligence technology has covered various stages of colorectal cancer, including screening, individualized assessment, auxiliary diagnosis and treatment decision-making, refined surgery and prognosis judgment, providing help for the accurate and individualized treatment of rectal cancer. However, the lack of standardized, systematic, and scalable AI models remains a major pain point for the field. Therefore, it is necessary to carry out large-scale prospective clinical studies on artificial intelligence model to further confirm its application value in the clinical diagnosis and treatment of rectal cancer.
6.Application value of artificial intelligence in surgical precision diagnosis and treatment of rectal cancer
Zhanwei FU ; Zhenghao CAI ; Junjun MA
Chinese Journal of Gastrointestinal Surgery 2024;27(6):574-578
Colorectal cancer is the most common malignant tumor of digestive tract, and the incidence of colorectal cancer in China is especially characterized by middle and low rectal cancer. In recent years, with the progress of computer science, artificial intelligence technology has developed rapidly, and has achieved a lot of application results in the medical field. At present, artificial intelligence technology has covered various stages of colorectal cancer, including screening, individualized assessment, auxiliary diagnosis and treatment decision-making, refined surgery and prognosis judgment, providing help for the accurate and individualized treatment of rectal cancer. However, the lack of standardized, systematic, and scalable AI models remains a major pain point for the field. Therefore, it is necessary to carry out large-scale prospective clinical studies on artificial intelligence model to further confirm its application value in the clinical diagnosis and treatment of rectal cancer.
7.Associations between abnormal electroencephalogram microstates and childhood emotional abuse in adolescent depression
Jinhui HU ; Dongdong ZHOU ; Lin ZHAO ; Lingli MA ; Xinyu PENG ; Xiaoqing HE ; Ran CHEN ; Wanjun CHEN ; Zhenghao JIANG ; Li KUANG ; Wo WANG
Journal of Central South University(Medical Sciences) 2023;48(11):1678-1685
Objective:Childhood traumatic experiences greatly influenced the brain network activities of patients with depression,and there is an urgent need to explore the temporal dynamics for these changes.This study aims to investigate the abnormalities of resting-state electroencephalogram(EEG)microstates in eye-open state of depressed adolescents and to explore the correlations between their EEG microstates and the childhood traumatic experience. Methods:Using resting-state EEG microstate analysis,we explored the temporal dynamics of brain activity in patients with adolescent depression.This study selected 66 adolescents with depression as a patient group,and 27 healthy adolescents as a healthy control group.A modified k-means clustering algorithm was used to classify the 64-channel resting-state EEG data into different microstates.Independent sample t-tests were used to compare the microstate parameters between the 2 groups and further assciations between these parameters and childhood traumatic experience in patients were explored via using Spearman correlation. Results:In this study,significant differences were observed in the occurrence and transition probabilities of EEG microstates between the healthy control and the patient group.Notably,there was a statistically significant difference(P<0.05)in the occurrence of microstate A across 2 groups,exhibiting a negative correlation with the emotional abuse component within the childhood trauma scores(Spearman's rho=-0.31,P=0.013).Furthermore,patient-specific,non-random transitions from microstate B to A(Spearman's rho=-0.30,P=0.015)and C to A(Spearman's rho=-0.31,P=0.013)were inversely associated with the scores of emotional abuse factors from childhood trauma in the patient group,showing statistically significant differences when comparing to the healthy controls(P<0.05).Upon stratification into quartiles based on the emotional abuse factor scores,the occurrence of microstate A,as well as the transition rates from microstates B to A and C to A,retained statistical significance following adjustment for multiple comparisons(all P<0.05). Conclusion:The abnormal temporal dynamics in brain networks of adolescents with depression are linked to childhood emotional abuse.Those who have suffered severe emotional abuse may show greater impairments in the brain's visual and central executive networks.EEG microstate analysis could be a potential tool for detecting adolescent depression with severe childhood trauma.
8.Clinical features of bone mass loss in liver cirrhosis and its association with sarcopenia
Qiu JIN ; Jing YANG ; Honglin MA ; Zhenghao SUN ; Qingqing LIU ; Shaoshan HU ; Pingju LI
Journal of Clinical Hepatology 2023;39(12):2831-2838
ObjectiveTo investigate the influence of sarcopenia on bone mass loss, the risk factors for bone mass loss in liver cirrhosis, and the correlation between body composition and bone mineral density (BMD) by comparing the clinical features of bone mass loss in patients with liver cirrhosis. MethodsA total of 92 patients who were hospitalized and diagnosed with liver cirrhosis in Department of Gastroenterology, The Second Affiliated Hospital of Kunming Medical University, from April to December of 2022 were enrolled, and based on the results of dual-energy X-ray absorptiometry, they were divided into bone mass loss group (osteopenia/osteoporosis) with 57 patients and normal bone mass group with 35 patients. The two groups were compared in terms of general data, laboratory examination, imaging data, and body composition analysis. The independent samples t-test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the chi-square test or the continuity correction chi-square test was used for comparison of categorical data between two groups; Pearson correlation analysis and Spearman correlation analysis were used to investigate correlation; a binary logistic regression analysis was used to investigate the risk factors for bone mass loss in liver cirrhosis. ResultsCompared with the normal bone mass group, the bone mass loss group had significantly higher age (t=-3.597, P<0.05), proportion of female patients (χ2=8.393, P<0.05), N-terminal middle molecular fragment of osteocalcin (N-MID) (Z=-3.068, P<0.05), β isomer of C-terminal telopeptide of type I collagen (β-CTX) (t=-2.784, P<0.05), and proportion of patients with sarcopenia (χ2=13.884, P<0.05) and significantly lower calcitonin (CT) (Z=-2.340, P<0.05) and L3 skeletal muscle index (L3-SMI) (t=4.621, P<0.05). Compared with the normal bone mass group, the bone mass loss group had significantly lower total muscle mass (Z=-2.952, P<0.05), right upper limb muscle mass (Z=-2.929, P<0.05), left upper limb muscle mass (Z=-2.680, P<0.05), right lower limb muscle mass (Z=-3.366, P<0.05), left lower limb muscle mass (Z=-3.374, P<0.05), presumed bone mass (t=2.842, P<0.05), body water mass (Z=-2.779, P<0.05), basal metabolic rate (BMR) (Z=-3.153, P<0.05), and BMD of L1— L4 and femoral neck (t=9.789, t=10.280, t=10.832, Z=-7.298, t=8.945, all P<0.05). Total muscle mass, muscle mass of trunk and limbs, presumed bone mass, BMR, and body water mass in body component analysis were positively correlated with L1 — L4 BMD and femoral neck BMD (all P<0.05), and fat mass was positively correlated with L1 — L4 BMD (all P<0.05). Sarcopenia (odds ratio [OR]=8.737, 95% confidence interval [CI]: 2.237 — 34.129, P=0.002), age (OR=1.094, 95%CI: 1.019 — 1.175, P=0.013), and N-MID (OR=1.095, 95%CI: 1.019 — 1.176, P=0.014) were independent risk factors for bone mass loss in patients with liver cirrhosis. ConclusionOld age, female sex, sarcopenia, elevated N-MID, elevated β-CTX, reduction in CT, low muscle mass, low presumed bone mass, low BMR, and low body water mass are the features of bone mass loss in patients with liver cirrhosis, and sarcopenia, age, and N-MID are independent risk factors for bone mass loss in patients with liver cirrhosis. Detailed assessment of body composition changes can help to identify abnormal BMD in patients with liver cirrhosis.
9.Clinical value of magnetic resonance imaging based integrated deep learning model for predic-ting the times of linear staplers used in middle-low rectal cancer resection
Zhanwei FU ; Zhenghao CAI ; Shuchun LI ; Luyang ZHANG ; Lu ZANG ; Feng DONG ; Minhua ZHENG ; Junjun MA
Chinese Journal of Digestive Surgery 2023;22(9):1129-1138
Objective:To investigate the clinical value of magnetic resonance imaging (MRI) based integrated deep learning model for predicting the times of linear staplers used in double stapling technique for middle-low rectal cancer resection.Methods:The retrospective cohort study was conducted. The clinicopathological data of 263 patients who underwent low anterior resection (LAR) for middle-low rectal cancer in Ruijin Hospital of Shanghai Jiaotong University School of Medicine from January 2018 to December 2022 were collected as training dataset. There were 183 males and 80 females, aged 63(55,68)years. The clinicopathological data of 128 patients with middle-low rectal cancer were collected as validation dataset, including 83 males and 45 females, with age as 65(57,70)years. The training dataset was used to construct the prediction model, and the validation dataset was used to validate the prediction model. Observation indicators: (1) clinicopathological features of patients in the training dataset; (2) influencing factors for ≥3 times using of linear staplers in the operation; (3) prediction model construction; (4) efficiency evaluation of prediction model; (5) validation of prediction model. Measurement data with skewed distribution were represented as M( Q1, Q3), and Mann-Whitney U test was used for comparison between groups. Count data were expressed as absolute numbers, and comparison between groups was conducted using the chi-square test. Wilcoxon rank sum test was used for non-parametric data analysis. Univariate analysis was conducted using the Logistic regression model, and multivariate analysis was conducted using the Logistic stepwise regression model. The receiver operating characteristic (ROC) curve was draw and the area under the curve (AUC) was calculated. The AUC of the ROC curve >0.75 indicated the prediction model as acceptable. Comparison of AUC was conducted using the Delong test. Results:(1) Clinicopathological features of patients in the training dataset. Of the 263 patients, there were 48 cases with linear staplers used in the operation ≥3 times and 215 cases with linear staplers used in the operation ≤2 times. Cases with preoperative serum carcinoembryonic antigen (CEA) >5 μg/L, cases with anastomotic leakage, cases with tumor diameter ≥5 cm were 20, 12, 13 in the 48 cases with linear staplers used ≥3 times in the operation, versus 56, 26, 21 in the 215 cases with linear staplers used ≤2 times in the operation, showing significant differences in the above indicators between them ( χ2=4.66, 5.29, 10.45, P<0.05). (2) Influencing factors for ≥3 times using of linear staplers in the operation. Results of multivariate analysis showed that preoperative serum CEA >5 μg/L and tumor diameter ≥5 cm were independent risk factors for ≥3 times using of linear staplers in the operation ( odds ratio=2.26, 3.39, 95% confidence interval as 1.15-4.43, 1.50-7.65, P<0.05). (3) Prediction model construction. According to the results of multivariate analysis, the clinical prediction model was established as Logit(P)=-2.018+0.814×preoperative serum CEA (>5 μg/L as 1, ≤5 μg/L as 0)+ 1.222×tumor diameter (≥5 cm as 1, <5 cm as 0). The image data segmented by the Mask region convolutional neural network (MASK R-CNN) was input into the three-dimensional convolutional neural network (C3D), and the image prediction model was constructed by training. The image data segmented by the MASK R-CNN and the clinical independent risk factors were input into the C3D, and the integrated prediction model was constructed by training. (4) Efficiency evaluation of prediction model. The sensitivity, specificity and accuracy of the clinical prediction model was 70.0%, 81.0% and 79.4%, respectively, with the Yoden index as 0.51. The sensitivity, specificity and accuracy of the image prediction model was 50.0%, 98.3% and 91.2%, respectively, with the Yoden index as 0.48. The sensitivity, specificity and accuracy of the integrated prediction model was 70.0%, 98.3% and 94.1%, respectively, with the Yoden index as 0.68. The AUC of clinical prediction model, image prediction model and integrated prediction model was 0.72(95% confidence interval as 0.61-0.83), 0.81(95% confidence interval as 0.71-0.91) and 0.88(95% confidence interval as 0.81-0.95), respectively. There were significant differences in the efficacy between the integrated prediction model and the image prediction model or the clinical prediction model ( Z=2.98, 2.48, P<0.05). (5) Validation of prediction model. The three prediction models were externally validated by validation dataset. The sensitivity, specificity and accuracy of the clinical prediction model was 62.5%, 66.1% and 65.6%, respectively, with the Yoden index as 0.29. The sensitivity, specificity and accuracy of the image prediction model was 58.8%, 95.5% and 92.1%, respectively, with the Yoden index as 0.64. The sensitivity, specificity and accuracy of the integrated prediction model was 68.8%, 97.3% and 93.8%, respectively, with the Yoden index as 0.66. The AUC of clinical prediction model, image prediction model and integrated prediction model was 0.65(95% confidence interval as 0.55-0.75), 0.75(95% confidence interval as 0.66-0.84) and 0.84(95% confidence interval as 0.74-0.93), respec-tively. There was significant differences in the efficacy between the clinical prediction model and the integrated prediction model ( Z=3.24, P<0.05). Conclusion:The MRI-based deep-learning model can help predicting the high-risk population with ≥3 times using of linear staplers in resection of middle-low rectal cancer with double stapling technique.
10.Effect of ubiquitination hepatitis B virus core antigen on inducing dendritic cells autophagy to enhance specific cytotoxic T lymphocyte responses
Run HUANG ; Jie CHEN ; Quanhui TAN ; Siyuan MA ; Xiaohua CHEN ; Yongsheng YU ; Guoqing ZANG ; Zhenghao TANG
Chinese Journal of Infectious Diseases 2021;39(4):228-233
Objective:To clarify the effect of ubiquitination hepatitis B virus core antigen (Ub-HBcAg) on dendritic cells (DC) autophagy, and to explore the mechanism of autophagy in enhancing DC antigen presentation and inducing hepatitis B virus-specific cytotoxic T lymphocyte (CTL) responses.Methods:Ub-HBcAg lentiviral vector (LV-Ub-HBcAg), lentiviral vector-hepatitis B virus core antigen (LV-HBcAg) and no-load plasmid LV (LV) were constructed and packaged. DC2.4 cells were divided into LV-Ub-HBcAg group, LV-HBcAg group and LV group. The blank control group (NC group) was also set. The protein expression of autophagy-related protein P62, microtubule associated protein 1 light chain 3 beta (LC3B), autophagy related 5(ATG5) and Beclin-1 were detected by Western blotting. The expressions of co-stimulatory molecules such as CD86, CD80 and major histocompatibility complex (MHC)-Ⅱ were detected by flow cytometry. Cell counting kit-8 (CCK-8) method was used to detect T lymphocytes proliferation. The non-radioactive lactic acid dehydrogenase (LDH) release method was applied to detect the killing ability of CTL. Statistical analysis was conducted by independent sample t test. Results:The relative protein expressions of LC3B-Ⅱ/LC3B-Ⅰ, Beclin-1 and ATG5 in NC group were 0.445±0.076, 0.522±0.026 and 0.761±0.038, respectively, which were all lower than those in LV-Ub-HBcAg group (0.926±0.021, 0.919±0.016 and 1.451±0.028, respectively). The relative protein expression of P62 in the NC group was higher than that in LV-Ub-HBcAg group ((1.875±0.016) vs (0.647±0.121)). The differences were all statistically significant ( t=6.102, 9.842, 17.490 and 10.590, respectively, all P<0.01). The expressions of CD86 (75.51%), CD80 (83.35%), MHC-Ⅱ (66.66%) in the LV-Ub-HBcAg group were high, and those in the NC group were 8.03%, 7.49%, 0.04%, respectively. The specific CTL killing rate ((65.310±2.091)%) of the LV-Ub-HBcAg group was significantly higher than both NC group ((14.400±0.497)%) and LV-HBcAg group ((54.870±1.443)%), and the differences were both statistically significant ( t=23.690 and 4.111, respectively, both P<0.05). Conclusion:Ub-HBcAg promotes the DC autophagy, up-regulates the expressions of costimulatory molecules on cell surface of DC to induce the maturation and activation, and then stimulates T lymphocyte to induce a stronger specific CTL response under the effort of ubiquitination.

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