Tumor microenvironment (TME) plays an important role in the structure and function of malignant tumor stroma. Cancer-associated fibroblasts (CAFs) have a strong ability of extracellular matrix synthesis and microenvironment remodeling, which can affect the development and progression of tumors. CAFs can produce the subgroups with different functions in different types of TME, and the study of CAF subgroups has a guiding significance for tumor therapy. This article reviews the effect of CAFs and its subgroups on the biological behavior and treatment of malignant tumors, in order to provide a reference for the clinical treatment of malignant tumors.

Objective To investigate the risk factors for delirium in elderly patients after artificial femoral head replacement for femoral neck fracture. Methods A total of 100 patients with femoral neck fracture who underwent artificial femoral head replacement in Department of Traumatology, The Affiliated Hospital of Qingdao University, from September 2019 to March 2022 were enrolled. Before surgery, State-Trait Anxiety Inventory was used to assess the status of anxiety, and after surgery, the Confusion Assessment Method was used to assess the condition of delirium. According to the presence or absence of delirium after surgery, the patients were divided into delirium group and non-delirium group. Univariate and multivariate logistic regression analyses were performed for the general data of the patients in both groups. Results The univariate logistic regression analysis showed that age, diabetes, coronary heart disease, body mass index, ASA score, Visual Analogue Scale score, and preoperative anxiety were significantly associated with the onset of postoperative delirium (POD) in eldevly patients (P<0.05). The multiva-riate logistic regression analysis showed that age (OR=1.194,95%CI=1.067-1.330,P<0.05), ASA score (OR=3.618,95%CI=1.075-12.177,P<0.05), and preoperative anxiety (OR=5.517,95%CI=1.098-27.710,P<0.05) were independent risk factors for postoperative delirium in elderly patients with femoral neck fracture. Conclusion Age, ASA score, and preope-rative anxiety are independent risk factors for the development of (POD) in elderly patients with femoral neck fracture. The assessment and intervention of these risk factors, especially the screening and management of preoperative anxiety, should be taken se-riously in clinical practice to reduce the incidence rate of (POD) and improve the prognosis of patients.

Objective To investigate the effect of remifentanil general anesthesia combined with thoracic paravertebral nerve block (TPVB) on the prognosis of elderly patients undergoing retroperitoneal laparoscopic radical nephrectomy. MethodsA total of 90 elderly patients who underwent retroperitoneal laparoscopic radical nephrectomy in our hospital from October 2024 to January 2025 were enrolled and divided into general anesthesia group (group C) and general anesthesia+TPVB group (group T) using the simple randomization method. The patients in group C were given with remifentanil general anesthesia alone, and those in group T underwent ultrasound-guided T₁₀ paravertebral nerve block at 30 minutes before surgery, followed by the same treatment in group C. The two groups were recorded and compared in terms of the amount of remifentanil and propofol used during surgery, awakening time from anesthesia, the length of stay in the postanesthesia care unit (PACU), Mini-Mental State Examination (MMSE) score before surgery and at 6, 24, and 48 hours after surgery, Visual Analogue Scale (VAS) score at 6, 12, 24, and 48 hours after surgery, 15-Item Quality of Recovery Scale (QoR-15) score at 24 hours after surgery, the dosage of sufentanil within 48 hours after surgery, time to the first pressing of patient-controlled intravenous analgesia (PCIA) within 48 hours after surgery, the proportion of patients requiring PCIA, the proportion of patients with remedial analgesia, and the proportion of patients with perioperative adverse reactions. Results Compared with group C, group T had a significantly lower proportion of patients with hypertension or tachycardia during surgery and a significantly lower amount of remifentanil used during surgery (χ²=12.17,14.95,t=5.80,P<0.05), as well as a significantly lower proportion of patients with bradycardia during surgery and a significant increase in QoR-15 score at 24 hours after surgery (χ²=6.43,t=-11.60,P<0.05). Both groups had a significant reduction in MMSE score at 6 hours after surgery (P<0.01), and MMSE score gradually increased over time after surgery (P<0.01). Group T had a signi-ficantly higher MMSE score than group C at 6 and 24 hours after surgery (P<0.01). Compared with group C, group T had significant reductions in the amount of sufentanil used at 48 hours after surgery, the proportion of patients requiring PCIA, the proportion of patients with remedial analgesia, the proportion of patients with postoperative delirium, awakening time, and the length of stay in the PACU (t=6.32-7.02,χ²=4.44-55.94,P<0.05), as well as a significantly longer time to the first pressing of PCIA within 48 hours after surgery (χ²=55.94,P<0.05). For both groups, VAS score at 12 hours after surgery were significantly hig-her than that at 6 hours after surgery, and VAS score gradually decreased with time since 12 hours after surgery (P<0.05). Group T had a significantly lower VAS score than group C at 6, 12, and 24 hours after surgery (P<0.01). Conclusion In elderly patients undergoing retroperitoneal laparoscopic radical nephrectomy, compared with remifentanil general anesthesia alone, its combination with TPVB can enhance postoperative analgesic effect and improve the quality of early postoperative recovery, with little impact on postoperative cognitive function, and it has no obvious adverse reactions.

Objective To evaluate the value of radiomic models based on contrast-enhanced computed tomography (CT) images in differentiating pancreatic ductal adenocarcinoma (PDAC) from pancreatic neuroendocrine neoplasms (pNEN). Methods A total of 291 cases of pancreatic tumors (218 PDAC cases and 73 pNEN cases) from The Affiliated Hospital of Qingdao University from November 2016 to September 2022 were assigned to the training set, while 124 cases (93 PDAC cases and 31 pNEN cases) from Shandong Provincial Hospital were assigned to the test set. Tumor regions of interest (ROI) were delineated on arterial-phase and venous-phase contrast-enhanced CT images of the upper abdomen, and subsequently, radiomic features were extracted and selected. Radom forest models were constructed using the radiomic features preprocessed with six methods separately: L1-norm regularization, L2-norm regularization, maximum absolute normalization, min-max normalization, quantile transformation, and Yeo-Johnson transformation. The efficacy of the six random forest models for distinguishing between PDAC and pNEN was assessed using the area under the receiver operating characteristic curve (AUC). The Brier score (BS) was calculated to evaluate model calibration. A decision curve analysis (DCA) was performed to quantify the net clinical benefit. Results For the six random forest models, the AUCs were 0.981-0.998 in the training set and 0.685-0.840 in the test set, with the accuracies being 92.8%-98.2% and 51.8%-86.7%, respectively. The quantile transformation-based random forest model demonstrated the best perfor-mance, showing an AUC of 0.840, a sensitivity of 90.2%, a specificity of 77.0%, and an accuracy of 86.7%, significantly surpassing those of the other models, also with the best calibration (BS=0.119). The DCA results indicated that the quantile transformation-based random forest model demonstrated superior net clinical benefits in both the training set and test set. Conclusion Radiomic models based on contrast-enhanced CT features can effectively diffe-rentiate PDAC from pNEN, in which the quantile transformation-based random forest model exhibits superior diagnostic performance, demonstrating potential for clinical application.

Objective To investigate the onset of neuropathic pain (NP) in patients with knee osteoarthritis (KOA) and related influencing factors. Methods A total of 313 KOA patients who attended the outpatient service of Department of Rehabi-litation Medicine in our hospital from July 2023 to October 2024 were enrolled, and according to the scoring results of the PainDETECT questionnaire, they were divided into NP group and non-NP group. The two groups were compared in terms of age, sex, body mass index (BMI), duration of knee pain symptoms, Kellgren-Lawrence (K-L) grading, pain Visual Analogue Scale (VAS) score, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), Hospital Anxiety and Depression Scale (HADS) score, and knee magnetic resonance imaging (MRI) findings. A logistic regression analysis was performed for the indicators with statistical significance identified by the univariate analysis to obtain the risk factors for NP. Results Of all patients, the patients with NP accounted for 16.61%. There were significant differences between the two groups in age, sex, pain VAS score, WOMAC, HADS score, and the proportion of patients with meniscus injury, soft tissue edema around the knee joint or knee joint ligament injury on MRI (H=16.272-49.953,χ²=12.897-23.792,P<0.05). The logistic regression analysis showed that old age, a high pain VAS score, a high HADS score, the presence of soft tissue edema around the knee joint, and the presence of knee joint ligament injury were risk factors for the onset of NP (P<0.05). Conclusion Old age, a high pain VAS score, a high HADS score, the presence of soft tissue edema around the knee joint, and the presence of knee joint ligament injury are risk factors for the onset of NP in KOA patients.

Objective To investigate the impact of single or continuous administration of subanesthetic doses of eske-tamine during anesthesia on postoperative depression in patients undergoing heart valve replacement. Methods A total of 256 patients who underwent heart valve replacement in our hospital from August 2023 to August 2024 were enrolled, and according to the use of esketamine during surgery, the patients were divided into control group (group C), single-dose group (group S-S), and continuous-dose group (group C-S). General information was collected and compared between the three groups, as well as 17-item Hamilton Depression Rating Scale (HAMD-17) score and the constituent ratio of patients with depressed mood on day 1 before surgery and on days 3, 7, and 30 after surgery, Visual Analog Scale (VAS) pain score and Self-Rating Scale of Sleep (SRSS) score on day 3 after surgery, and the incidence rate of adverse reactions within 1 week after surgery. Results Group C-S had a significantly lower VAS score than group C on day 3 after surgery (F=4.617,P<0.05). The three groups showed a significant change in HAMD-17 score from before surgery to different time points after surgery (F_intragroup=4.497-18.254,P<0.05); in group C, HAMD-17 score on day 3 after surgery was significantly higher than that on day 1 before surgery (P<0.01), while in group C-S, HAMD-17 score on day 7 after surgery was significantly higher than that on day 1 before surgery (P<0.05); for all three groups, HAMD-17 score at 1 month after surgery was significantly lower than that on day 1 before surgery (P<0.05). There were significant diffe-rences in HAMD-17 score between the three groups on days 3 and 7 after surgery (F_intergroup=7.218,7.809,P<0.05); groups C-S and S-S had a significantly lower HAMD-17 score than group C on day 3 after surgery (P<0.05), while group C-S had a significantly lower HAMD-17 score than group C on day 7 after surgery (P<0.05). For group C, the constituent ratio of patients who deve-loped depressed mood on day 3 after surgery was significantly higher than that on day 1 before surgery (Wald χ²=33.704,P<0.01), and there was no significant change in the constituent ratio of patients who developed depressed mood after surgery in the other two groups (P>0.05). Conclusion Continuous intrao-perative application of subanesthetic doses of esketamine can improve depressive symptoms within a short period of time after surgery and alleviate postoperative pain in patients undergoing heart valve replacement, without increasing the incidence rate of adverse reactions. This approach can help to promote the postoperative recovery of patients undergoing heart valve replacement.

Objective To establish and validate a model for predicting the SSIGN score and prognosis of patients with clear cell renal cell carcinoma (ccRCC) based on CT radiomic features and clinical indices. Methods A total of 845 patients with pathologically confirmed ccRCC from June 2013 to July 2022 were enrolled, with 592 patients in the training set and 253 in the test set, and according to the SSIGN score, the patients were divided into intermediate-to-high recurrence risk group (≥4 points) and low recurrence risk group (0-3 points). The clinical indices and CT radiomic features of ccRCC were collected from all patients, and univariate and multivariate logistic regression analyses were used to identify the influencing factors for SSIGN score. A predictive model for SSIGN score was established based on CT radiomic features and clinical indices. The receiver operating characteristic (ROC) curve and the area under the ROC curve (AUC) were used to evaluate the performance of the model in both the training set and the test set. The decision curve analysis was used to assess the net clinical benefit of the model in the test set. The Kaplan-Meier (K-M) survival analysis was used to assess the correlation of SSIGN score predicted by the model with recurrence-free survival (RFS) in both the training set and the test set, and index of concordance (C-index) and hazard ratio (HR) were used to eva-luate the performance of the model in predicting RFS. Results In the training set, this model had an AUC of 0.93 in predicting SSIGN score in ccRCC patients, while it had an AUC of 0.92 in the test set. The calibration curves showed that the combined model had high calibration ability for predicting SSIGN score in both the training set and the test set, and the DCA results showed that the combined model had a relatively high net clinical benefit in the test set. The K-M survival curve analysis showed a significant difference in RFS between the intermediate-to-high recurrence risk group and the low recurrence risk group based on SSIGN score predicted by the combined model (χ²=104.718,82.229,P<0.001). The mo-del had a C-index of 0.76 and an HR of 40.51 in the training set, as well as an C-index of 0.77 and an HR of 185.27 in the training set. Conclusion The combined model based on CT radiomic features and clinical indices shows relatively high predictive efficacy for the SSIGN score and prognosis of ccRCC patients. As a novel noninvasive assessment tool, this model has a certain potential for clinical application in the prognostic evaluation of ccRCC patients.

Objective To establish a combined diagnostic model based on CT radiomics and clinical features, and to investigate the value of the model in the diagnosis of aldosterone-producing adenoma (APA) of the adrenal gland. Methods This study was conducted among 229 patients with a unilateral solitary adrenal mass who were admitted to The Affiliated Hospital of Qingdao University from January 2018 to December 2023 and 50 patients with a unilateral solitary adrenal mass who were admitted to Qilu Hospital of Shandong University from January 2016 to March 2024, and according to whether they were diagnosed with APA, each batch of patients was divided into APA group and non-APA group. Based on the time-series segmentation method, the 229 patients (104 in the APA group and 125 in the non-APA group) from The Affiliated Hospital of Qingdao University were divided into a training set (n=160) and an internal validation set (n=69) according to the order of operation time at a ratio of 7∶3, and the 50 patients (18 in the APA group and 32 in the non-APA group) from Qilu Hospital of Shandong University were set as an external validation group. Clinical and imaging data were collected from all patients, and univariate and multivariate logistic regression analyses were used to investigate the independent influencing factors for the onset of unilateral solitary APA and establish a clinical feature model based on these factors. The Pearson correlation coefficient and the least absolute shrinkage and selection ope-rator (LASSO) algorithm were used to identify the radiomic features on the plain CT and contrast-enhanced CT images of the adrenal gland, and a CT radiomic model was established. The indepen-dent influencing factors and the radiomic features were used to establish a combined diagnostic model for unilateral solitary APA, and a nomogram was plotted. The receiver operating characteristic (ROC) curve and the area under the ROC curve (AUC) were used to evaluate the performance of the model, and the clinical decision analysis (DCA) and the calibration curve were used to assess the clinical application value of the nomogram. Results The univariate and multivariate logistic regression analyses showed that preoperative serum potassium level, the maximum diameter of the lesion, and the highest CT value (plain scan) of the lesion were independent influencing factors for the onset of unilateral solitary APA, and a clinical feature model was established based on these three indicators, while the 23 radiomic features identified by plain CT scan of the adrenal gland were used to establish a radiomic model. A combined diagnostic model was established based on the independent influencing factors and the radiomic features and was represented by a nomogram, and the results showed that it had good diagnostic efficacy. In the training set, the clinical model, the CT radiomic model, and the combined diagnostic model had an AUC of 0.761, 0.867, and 0.949, respectively, in the internal validation group, they had an AUC of 0.733, 0.718, and 0.895, respectively, and in the external validation group, they had an AUC of 0.739, 0.716, and 0.929, respectively, suggesting that the combined diagnostic model had a significantly greater AUC than the other two models in the training group, the internal validation group, and the external validation group (Z=2.495-5.327,P<0.05). The DCA showed that the combined diagnostic model had the maximum net clinical benefit, and the calibration curve showed that the combined diagnostic model had good calibration. Conclusion In this study, a diagnostic model is established for unilateral solitary APA based on clinical and radiomic features, which has excellent diagnostic efficacy and a good generalization ability for APA, and therefore, it is expected to become an ideal noninvasive diagnostic method for unilateral solitary APA in clinical practice.

Objective To investigate the clinicopathological significance of the Asian Cancer Research Group (ACRG) classification in gastric adenocarcinoma and to develop a predictive model for p53 protein expression in gastric adenocarcinoma. Methods A total of 137 patients diagnosed with gastric adenocarcinoma who underwent surgery at the Affiliated Hospital of Qingdao University between 2015 and 2023 were included in this study. Postoperative pathological specimens were collected and immunohistochemically stained to determine the ACRG classification. Patients were grouped according to ACRG subtypes. These groups were compared in terms of age, sex, surgical approach, tumor location, differentiation status, pathological type, pre-chemotherapy Borrmann classification, clinical stage, tumor marker levels, tumor recurrence, and metastasis. Preoperative hematoxylin and eosin-stained endoscopic biopsy slides from all patients were digitized by scanning and randomly divided into a training set (109 cases) and a validation set (28 cases). Self-supervised learning and feature extraction were performed on the training set images using SimCLR with ResNet-18 as the backbone network. Subsequently, the images were input into a feature aggregation-based classification network to establish a predictive model for p53 protein expression in gastric adenocarcinoma. The model performance was eva-luated in the validation set. Receiver operating characteristic curves were plotted for both the training and validation sets, and the area under the curve, accuracy, sensitivity, specificity, positive predictive value, and negative predictive value were calculated. Results The ACRG classification of gastric adenocarcinoma was significantly associated with clinical stage (χ²=13.049,P<0.05). In the training set, the area under the curve, accuracy, sensitivity, specificity, positive predictive value, and negative predictive value of the p53 protein expression prediction model were 0.920, 0.719, 0.896, 0.724, 0.875, and 0.763, respectively; these values in the validation set were 0.890, 0.643, 0.938, 0.250, 0.625, and 0.750, respectively. Conclusion The ACRG classification of gastric adenocarcinoma is significantly correlated with clinical stage. The p53 protein expression prediction model can assist in preoperative ACRG classification, thereby improving the accuracy of diagnosis and treatment of gastric adenocarcinoma.

Objective To investigate the risk factors for otitis media and sinusitis in patients with nasopharyngeal carcinoma after radiothe-rapy. Methods The clinical data from 207 patients with nasopharyngeal carcinoma who underwent radiotherapy at the Affiliated Hospital of Qingdao University between January 2013 and December 2019 were collected. The clinical characteristics of patients who developed otitis media and sinusitis after radiotherapy were compared with those of patients who did not. Univariate and multivariate logistic regression analyses were performed to identify risk factors for post-radiotherapy otitis media and sinusitis. Additionally, the correlation between the occurrence of otitis media and sinusitis after radiotherapy was analyzed. Results Comparison of 207 patients with nasopharyngeal carcinoma before and after radiotherapy showed statistically significant differences in the severity of otitis media and sinusitis (χ²=12.903-31.373,P<0.05). Multivariate logistic regression analysis revealed that tumor T stage was a risk factor for post-radiotherapy otitis media (P<0.05), and smoking and tumor T stage were risk factors for post-radiotherapy sinusitis (P<0.05). A significant correlation was observed between the occurrence of post-radiotherapy otitis media and sinusitis (r=0.22,P<0.05). Conclusion For patients with nasopharyngeal carcinoma at a high T stage and having a history of smoking, close attention should be paid to the occurrence of post-radiotherapy otitis media and sinusitis. Early intervention is essential to improve patients’ quality of life.

Objective To establish an indirect ELISA method for the detection of human cytomegalovirus (HCMV) based on HCMV IE1/pp65/pp150 recombinant protein. Methods From June to December 2024, 5 serum samples from HCMV IgG-positive patients, 50 serum samples from HCMV IgG-negative patients, as well as 8 serum samples each from the patients with positive herpes simplex virus-1 (HSV-1), herpes simplex virus-2 (HSV-2), and Epstein-Barr virus (EBV), and 349 serum samples from volunteers undergoing physical examination were collected from Qingdao Municipal Hospital. An indirect ELISA method for HCMV was established by optimizing the detection conditions such as serum dilution ratio and the type of blocking solution, and the cut-off value of this method was determined; coefficient of variation (CV) was calculated to verify the repeatability of the method; the sensitivity and specificity of the method were assessed by detecting the serum samples from the patients with positive HCMV IgG, HSV-1, HSV-2, and EBV at various dilution ratios; the consistency rate between HCMV indirect ELISA and cytomegalovirus IgG ELISA kit was assessed by analyzing the serum samples of 349 healthy volunteers undergoing physical examination at the outpatient service. Results The optimal antigen-coating concentration of HCMV indirect ELISA assay was 3.0 mg/L, with an optimal serum dilution ratio of 1∶200, an optimal blocking solution of 5% bovine serum albumin, an optimal blocking time of 2 h, an optimal serum incubation time of 90 min, an optimal dilution ratio of the second antibody of 1∶8 000, an optimal incubation time of the second antibody of 30 min, and an optimal color development time of 15 min. The cut-off value of HCMV indirect ELISA assay was 0.360, with an intra- or inter-batch CV of <10% and a sensitivity of 1∶3 200, and the absorbance values of serum samples from HSV-1, HSV-2, and EBV-positive patients were all below the cut-off value. The overall consistency rate between HCMV indirect ELISA and cytomegalovirus IgG ELISA was 94.0%. Conclusion An indirect ELISA method is successfully established for detecting HCMV with HCMV IE1-pp65-pp150 recombinant protein as the coating antigen, and this method has good repeatability, specificity, sensitivity, and consistency rate.

Objective To investigate the protective effect and mechanism of (-)-(S)-B-973B, an allosteric modulator of α7 nicotinic acetylcholine receptor, on SH-SY5Y cell injury induced by 1-methyl-4-phenyl pyridine (MPP⁺). Methods CCK-8 assay was used to investigate the effect of 0, 100, 200, 300, 500, 1 000, 2 000 μmol/L MPP⁺ on the viability of SH-SY5Y cells (groups A1-G1, respectively), the effect of 0, 0.1, 1, 3, 10, 30 and 100 μmol/L (-)-(S)-B-973B on the viability of SH-SY5Y cells (groups A2-G2, respectively), and the effect of 0, 0.1, 1, and 3 μmol/L (-)-(S)-B-973B on the viability of SH-SY5Y cells induced by MPP⁺ (groups B3-E3, respectively). SH-SY5Y cells were divided into control group (group A), MPP⁺ group (group B), (-)-(S)-B-973B group (group C), (-)-(S)-B-973B+MPP⁺ group (group D). Flow cytometry was used to observe the effect of on mitochondrial transmembrane potential in groups A-D cells. Western blotting was used to measure the expression le-vel of the superoxide dismutase 1 (SOD1) and Bcl-2/Bax ratio in groups A-D cells. Results CCK-8 assay showed that compared with group A1, groups C1-G1 had a significant reduction in cell viability (F=11.11,t=3.60-6.91,P<0.05); compared with group A2, the groups E2-G2 had a significant reduction in cell viability (F=131.90,t=3.26-20.35,P<0.05), suggesting that (-)-(S)-B-973B with a concentration of above 10 μmol/L had significant toxicity to SH-SY5Y cells; compared with group B3, group D3 had a significant increase in cell viability (F=20.49,t=2.41,P<0.05). Flow cytometry showed that compared with group A, group B had a significant reduction in mitochondrial transmembrane potential (F=4.51,t=2.98,P<0.05), and compared with group B, group D had a significant increase in mitochondrial transmembrane potential (t=3.36,P<0.05). Western blotting showed that compared with group A, group B had a significant reduction in the protein expression level of SOD1 (F=4.61,t=4.19,P<0.05), and compared with group B, group D had a significant increase in the protein expression level of SOD1 (t=2.89,P<0.05); there was no significant difference in Bcl-2/Bax ratio between groups B-D (P>0.05). Conclusion This study shows that (-)-(S)-B-973B exerts a protective effect against SH-SY5Y cell injury induced by MPP⁺, possibly by enhancing mitochondrial function and exerting an anti-oxidative stress effect.

Objective To investigate the regulatory effect of Epstein-Barr virus (EBV) on the expression of sterile alpha and Toll/interleukin receptor motif-containing protein 1 (SARM1) and its effects on the proliferation and migration of gastric can-cer cells. Methods From the GSE51575 dataset of the Gene Expression Omnibus, we obtained the relative mRNA expression levels of the SARM1 gene of 12 cases of EBV-negative gastric cancer and 12 cases of EBV-positive gastric cancer. The expression data of the SARM1 gene were analyzed using the online GEO2R tool. AGS cells were divided into AGS group and AGS+12-O-tetradecanoylphorbol-13-acetate (TPA) group, and AGS-EBV cells were divided into AGS-EBV group and AGS-EBV+TPA group. The AGS+TPA group and AGS-EBV+TPA group were treated with TPA at 20 μg/L for 24 h, while the AGS group and AGS-EBV group were treated with the same volume of dimethyl sulfoxide for 24 h. The culture medium supernatant of the AGS+TPA group and AGS-EBV+TPA group was collected; and AGS cells were divided into AGS supernatant group and AGS-EBV supernatant group to be cultured with the AGS+TPA cell culture medium supernatant and the AGS-EBV+TPA cell culture medium supernatant for 5 d, respectively. Si-NC group, si-SARM1-1 group, and si-SARM1-2 group were set with AGS cells; AGS-EBV-NC group and AGS-EBV-SARM1 group were set with AGS-EBV cells; and YCCEL1-NC group and YCCEL1-SARM1 group were set with YCCEL1 cells. The si-NC group, si-SARM1-1 group, and si-SARM1-2 group were transfected with si-NC, si-SARM1-1, and si-SARM1-2, respectively; the AGS-EBV-NC group and YCCEL1-NC group were transfected with control plasmids; and the AGS-EBV-SARM1 group and YCCEL1-SARM1 group were transfected with the SARM1 recombinant expression plasmid. The expression of SARM1 was measured by proteomic sequencing and Western blot. Cell absorbance was measured by using Cell Counting Kit-8. The relative cell migration rate was determined by Transwell migration assay. Results Compared with those in EBV-ne-gative gastric cancer tissues and EBV-negative gastric cancer cell lines (HGC27, MGC803, and AGS), the relative expression le-vels of SARM1 mRNA and protein in EBV-positive gastric cancer tissues and EBV-positive gastric cancer cell lines (YCCEL1, GT38, GT39, SNU719, and AGS-EBV) were significantly lower (F=101.70,t=2.77-69.28,P<0.05). Compared with the AGS-EBV group and AGS supernatant group, the AGS-EBV+TPA group and AGS-EBV supernatant group showed significantly lower relative expression levels of SARM1 protein (t=6.99,93.13,P<0.05). There were significant differences in the absorbance value, re-lative cell migration, and the relative expression levels of SARM1, ZEB1, and β-catenin between the si-NC group and the si-SARM1-1 group and si-SARM1-2 group, between the AGS-EBV-NC group and the AGS-EBV-SARM1 group, and between the YCCEL1-NC group and the YCCEL1-SARM1 group (F=22.70-2 191.00,q=3.85-22.39,t=5.34-26.49,P<0.05). Conclusion The expression of SARM1 is significantly decreased in EBV-positive gastric cancer cells. EBV may inhibit the proliferation and migration of gastric cancer cells by lowering the expression of SARM1.

Objective To explore the effect of the transcriptional coactivator with PDZ-binding motif (TAZ) on the proliferation and migration of breast cancer cells and its molecular mechanism. Methods Human breast cancer cells MCF7 and BT549 were cultured in vitro and divided into groups A-J and a-j, respectively. The cells were transfected with pcDNA3.1, TAZ, PLKO.1, shTAZ, TAZ+PLKO.1, TAZ+shGLUT3, WT pGLUT3+pcDNA3.1, WT pGLUT3+TAZ, pGLUT3 mutation+pcDNA3.1, and pGLUT3 mutation+TAZ. Colony-forming assay (CFA) and Transwell assay were used to determine plating efficiency and migration rate in groups A-F and a-f. Quantitative reverse transcription PCR (RT-qPCR) was used to measure the relative expression of GLUT3 mRNA downstream of TAZ in groups A-D and a-d. The GLUT3 protein expression level in groups A-F and a-f was measured using Western blotting (WB). The binding site of TAZ transcription enhanced association domain (TEAD) and GLUT3 was predicted using the JASPER database, and the relative luciferase activity of cells in groups G-J and g-j was measured using dual luciferase reporter (DLR) assay. Results The results of the CFA showed that the plating efficiency was significantly higher in groups B/b than in groups A/a (t=23.04,25.97,P<0.05), and significantly lower in groups D/d and F/f than in groups C/c and E/e (t=9.76-42.68,P<0.05). The results of the Transwell assay showed that the cell migration rates were significantly higher in groups B/b than in groups A/a (t=30.85,29.44,P<0.05), and significantly lower in groups D/d and F/f than in groups C/c and E/e (t=12.40-33.08,P<0.05). The results of the DLR assay showed that the relative luciferase activity was significantly higher in groups H/h than in groups G/g (F=138.73,222.10,t_LSD=11.09,25.81,P<0.05). The RT-qPCR results showed that the relative expression levels of GLUT3 mRNA were significantly higher in groups B/b than in groups A/a (t=15.80,14.53,P<0.05), and significantly lower in groups D/d than in groups C/c (t=4.33,4.11,P<0.05). The results of the WB showed that the relative expression levels of GLUT3 were significantly higher in groups B/b than in groups A/a (t=2.22,6.50,P<0.05), and significantly lower in groups D/d and F/f than in groups C/c and E/e (t=2.67-8.00,P<0.05). Conclusion TAZ may bind to the GLUT3 promoter region through TEAD to upregulate GLUT3 expression, thus promoting the proliferation and migration of breast cancer cells.

Objective To investigate the effect of the photothermal agent Cy7-TCF on oxidative stress and apoptosis in human pancreatic cancer cells. Methods Mia PaCa-2 cells were divided into control group, Cy7-TCF group (treated with Cy7-TCF alone), and Cy7-TCF+laser group (treated with Cy7-TCF followed by 808 nm laser irradiation). CCK-8 assay was used to measure cell viability under different concentrations of Cy7-TCF with or without laser irradiation; fluorescence microscopy was used to observe the uptake of Cy7-TCF by cells, and confocal microscopy combined with a lysosomal probe was used to observe the subcellular localization of Cy7-TCF; the DCFH-DA fluorescent probe was used to measure the level of reactive oxygen species (ROS) in cells; flow cytometry was used to measure cell apoptosis rate. Results CCK-8 assay showed that under laser irradiation, cell viability significantly decreased with the increasing concentration of Cy7-TCF in a concentration-dependent manner (P<0.05). Fluo-rescence microscopy showed that Cy7-TCF was effectively taken up by Mia PaCa-2 cells, and confocal microscopy showed that Cy7-TCF was mainly localized within lysosomes. DCFH-DA probe detection showed that under 808 nm laser irradiation, Cy7-TCF significantly increased the level of ROS in Mia PaCa-2 cells (q=19.07,16.84,P<0.05). Flow cytometry showed that the Cy7-TCF+laser group had a significantly higher apoptosis rate than the control group and the Cy7-TCF group (q=28.78,24.89,P<0.05). Conclusion Cy7-TCF is effectively taken up by Mia PaCa-2 cells and is localized in lysosomes, and through its photothermal effect, it induces the increase in the level of ROS in cells, reduces cell viability, and promotes cell apoptosis.

Objective To investigate the effect of the TTC-MMP12_184-225 vaccine, which is derived from matrix metalloproteinase 12 (MMP12) and tetanus toxin C-fragment (TTC), for the treatment of lung cancer and the underlying mechanisms. Methods The Tumor Immune Estimation Resource (TIMER) database was used to analyze the relative mRNA expression of MMP12 in the tumor tissues and adjacent normal tissues of patients with lung cancer. A subcutaneous tumor model of lung cancer was established by subcutaneously inoculating Lewis lung carcinoma (LLC) cells at 2-3 cm below the right axilla of 6-8 week-old C57BL/6J female mice. When the tumor volume reached approximately 100 mm³, the lungs and tumors were isolated to determine the mRNA expression of MMP12 by RT-qPCR. The CD8⁺T cell epitopes, CD4⁺T cell epitopes, and B cell epitopes of MMP12 were predicted on the NetCTL-1.2, SYFPEITHI, and SVMTrip websites, respectively. The immunogenicity of the dominant epitopes was analyzed using the VaxiJen-2.0 server. The secondary structure and hydrophilicity of MMP12 were analyzed by using DNAstar software. TTC-MMP12_184-225 protein was synthesized through the Escherichia coli prokaryotic expression system, and the product was identified with Coomassie blue staining. Subcutaneously inoculated with LLC cells at 2-3 cm below the right axilla, a group of 6-8 week-old C57BL/6J female mice were randomized to receive intramuscular injection of 100 μL PBS containing either 20 μg TTC (control group) or 20 μg TTC-MMP12_184-225 (experimental group), once every 7 d for a total of 3 times. On the third day after the last injection, the tumors were isolated for volume measurement; the proportions of CD4⁺T cells and CD8⁺T cells in tumors were determined by flow cytometry; and the relative mRNA expression levels of C-C motif chemokine ligand 5 (CCL5), C-X-C motif chemokine ligand 9 (CXCL9), interleukin-2 (IL-2), interferon gamma (IFN-γ), granzyme B (GRZB), and perforin (PFP) in tumors were measured by RT-qPCR. Results The TIMER and RT-qPCR results showed that the expression of MMP12 was significantly increased in both patients with lung cancer and the mouse model of lung cancer compared with adjacent normal tissues and lung tissues (P<0.001、0.05). The epitope prediction results showed that MMP12_184-225 contained 12 CD8⁺T cell epitopes, 9 CD4⁺T cell epitopes, and 3 B cell epitopes. The VaxiJen-2.0 analysis results showed that the immunogenicity score of MMP12_184-225 was 0.61. The DNAstar analysis showed that MMP12_184-225 was composed of hydrophilic peptides mainly consisting of β-turns. The Coomassie blue staining results showed that TTC-MMP12_184-225 protein exhibited a clear band below the relative molecular mass of 60 000. Compared with those in the control group, the mice in the experimental group showed a significantly reduced tumor volume, significantly increased proportions of CD4⁺T cells and CD8⁺T cells in tumors, and significantly increased relative expression levels of CCL5, CXCL9, IL-2, IFN-γ, GRZB, and PFP mRNA in tumors (t=2.57-4.60,P<0.05). Conclusion The TTC-MMP12_184-225 vaccine exerts an anti-lung cancer effect by promoting the recruitment of CD4⁺T cells and CD8⁺T cells and increasing the expression of antitumor-related cytokines in tumor tissues.

Objective To investigate the effect and mechanism of sodium butyrate (NaB) on the activation of Epstein-Barr virus (EBV) in EBV-positive gastric cancer cells. Methods EBV-positive gastric cancer cells (AGS-EBV cells) were divi-ded into groups A, B, and C, and the cells in group A were cultured routinely for 12 hours, while those in groups B and C were treated with NaB for 12 and 24 hours respectively. Transcriptome sequencing was performed for the genes of AGS-EBV cells and the EBV genes, and groups A, B, and C were compared in terms of the relative expression levels of key latent genes and key lytic genes of EBV in cells. Differentially expressed genes (DEGs) were identified between groups A, B, and C, and the top 5 upregula-ted DEGs and the top 5 downregulated DEGs with smaller P values were considered significant DEGs, while RT-qPCR was used to measure the relative expression levels of significant DEGs. The GO functional enrichment analysis and the KEGG pathway enrichment analysis were performed for all DEGs identified, and the proteins enriched in significant pathways (the pathways with P<0.05 in the KEGG pathway enrichment analysis) were considered key pathway proteins, while Western blotting was used to mea-sure the relative expression levels of key pathway proteins in groups A, B, and C. The DEGs in groups A, B, and C were compared with the transcription factors in the AnimalTFDB/PlantTFDB database, and the top 5 transcription factors with smaller P values were selected as core transcription factors, while RT-qPCR was used to measure the relative expression levels of core transcription factors in groups A, B, and C. Results Compared with group A, groups B and C had significant increases in the relative expression le-vels of key latent genes (EBNA-1, EBNA-3A, EBNA-3B, EBNA-3C, EBNA-LP, LMP-1, LMP-2A, LMP-2B and RPMS1) and lytic period genes (BZLF1, BRLF1 and BHLF1) (t=2.67-14.36,P<0.05), and group C had significantly higher expression levels of these genes than group B (t=2.21-6.97,P<0.05). Compared with group A, group B had significant increases in the relative expression levels of SCN1A, RFPL4AL1, RORB, and CACNG7 in cells (t=2.81-13.93,P<0.05) and significant reductions in the relative expression levels of MIR205HG, PLAAT4, and OAS2 (t=4.47-623.30,P<0.05); compared with group A, group C had significant increases in the relative expression levels of NLRP5, SCN1A, RFPL4AL1, RORB, CACNG7, and PLA2G2E (t=4.32-7.43,P<0.05) and significant reductions in the relative expression levels of RS17P11, MIR205HG, PLAAT4, and OAS2 (t=3.44-1 510.00,P<0.05). Compared with group A in terms of the core transcription factors, groups B and C had significant increases in the relative mRNA expression levels of EGR1, PGR, and SOX2 (t=3.22-16.60,P<0.05) and significant reductions in the relative mRNA expression levels of CEBPD and E2F3 (t=4.18-28.59,P<0.05). Compared with group A, group B had significant reductions in the relative expression levels of p65, p-p65, AKT, and p-AKT (t=2.87-5.93,P<0.05), and compared with group A, group C had significant reductions in the relative expression levels of p65, p-p65, AKT, and p-AKT and a significant increase in the expression level of p-ERK (t=5.72-8.92,P<0.05). Conclusion NaB promotes the activation of EBV in EBV-positive gastric cancer cells by activating the MAPK signaling pathway and inhibiting the NF-κB pathway, and the core transcription factor EGR1 promotes the activation of the virus by facilitating the transcription of the EBV lytic genes BZLF1 and BRLF1.

Real-time shear wave elastography (SWE) is a new elastography technique applied in clinical practice,with the advantages of noninvasiveness,repeatable operation,and stable accuracy, and it can be used for real-time dynamic monitoring and evaluation of liver fibrosis degree in liver diseases.This article reviews the imaging principle of SWE and its application in the diagnosis of liver diseases,in order to provide reliable information for the early diagnosis,treatment,and efficacy evaluation of liver di-seases.

Objective To investigate the value of gemstone spectral CT in evaluating the differentiation degree of lung squamous cell carcinoma. Methods A total of 86 patients with lung squamous cell carcinoma who were diagnosed and treated in Puyang People’s Hospital from February 2019 to February 2020 were enrolled as subjects, and according to the results of postope-rative pathological examination, they were divided into group A (poorly differentiated), group B (moderately differentiated), and group C (highly differentiated). These three groups were compared in terms of gemstone spectral CT imaging parameters and di-sease conditions in the venous and arterial phases. The receiver operating characteristic (ROC) curve was plotted to investigate the efficacy of various gemstone spectral CT imaging parameters in assessing the differentiation degree of lung squamous cell carcinoma. Results There were no significant differences in gemstone spectral CT imaging parameters in the venous phase between groups (P>0.05). Compared with group C, groups A and B had significantly lower gemstone spectral CT imaging parameters of iodine concentration (IC), normalized iodine concentration (NIC), slope K value and effective atomic coefficient in the arterial phase, and group A had significantly lower parameters than group B (t=2.322-14.746,P<0.05). The ROC curve analysis showed that the combination of IC, NIC, slope K value, and effective atomic coefficient had the highest specificity and area under the ROC curve in evaluating the differentiation degree of lung squamous cell carcinoma. Conclusion The parameters of gemstone spectral CT have a relatively high value in assessing the differentiation degree of arterial-phase lung squamous cell carcinoma.

Objective To investigate the effect of outpatient or home-based cardiac rehabilitation training on cardiopulmonary function and mental health after percutaneous coronary intervention (PCI) in young and middle-aged patients with coronary heart disease. Methods A total of 110 patients who underwent PCI in our hospital from January 2020 to December 2021 were randomly divided into home-based group with 56 patients and outpatient group with 54 patients. An individualized rehabilitation training prescription was formulated based on the results of CPET assessment, and the patients in the outpatient group received cardiac rehabilitation training at the outpatient service, while those in the home-based group received cardiac rehabilitation training at home. Related data were recorded before training and after 3 months of training, including cardiorespiratory exercise test results, 7-item Generalized Anxiety Disorder Scale (GAD-7) score, and Patient Health Questionnaire-9 (PHQ-9) score, and the changes in these indicators were compared between the two groups. Results After rehabilitation training, both groups had significant increases in anaerobic threshold power, anaerobic threshold metabolic equivalent (ATMets), anaerobic threshold oxygen uptake per kilogram (ATVO₂), peak power (PEAKPower), peak oxygen uptake per kilogram (PEAKVO₂max), and peak metabolic equivalent (PEAKMets) (t=2.22-6.43,P<0.05), with no significant changes in heart rate, systolic blood pressure, and diastolic blood pressure at resting state (P>0.05), and there were significant differences between the two groups in the changes in ATMets, ATVO₂, PEAKPower, PEAKMets, and PEAKVO₂max after training (z=2.23-4.10,P<0.05). Both groups had significant reductions in GAD-7 and PHQ-9 scores after rehabilitation training (t=5.82-8.59,P<0.05), and there were no signi-ficant differences in the changes in GAD-7 and PHQ-9 scores between the two groups after training (P>0.05). Conclusion Both outpatient and home-based cardiac rehabilitation training can improve cardiopulmonary function and alleviate anxiety and depression after PCI in young and middle-aged patients with coronary heart disease, and outpatient cardiac rehabilitation training has a better effect than home-based training.

Objective To investigate the risk factors for the onset of glioma in patients with the initial symptom of epilepsy. Methods A total of 309 patients with glioma who were diagnosed and treated in our hospital from January 2016 to December 2019 were enrolled, and according to whether the initial symptom was epilepsy, they were divided into observation group with 81 patients and control group with 228 patients. A retrospective analysis was performed for general information (age, sex, smoking history, drinking history, history of chronic diseases and head trauma, extent of tumor lesion, and degree of tumor diffe-rentiation) and blood biochemical parameters (total cholesterol, triglycerides, high-density/low-density lipoprotein cholesterol, fasting blood glucose, calcium, potassium, and phosphorus), and a multivariate logistic regression analysis was used to investigate the risk factors for epileptic seizures in patients with glioma. Results Among the patients with glioma, the patients with epileptic seizures as the initial symptom accounted for 18.95%. The analysis showed that there were significant differences between the two groups in the history of drinking, epilepsy, diabetes, and head trauma, the extent of tumor lesion, the degree of tumor diffe-rentiation, and the serum levels of total cholesterol, low-density lipoprotein cholesterol, fasting blood glucose, potassium, and phosphorus (χ²=3.915-12.513,P<0.05). The multivariate logistic regression analysis showed that the history of drinking/epilepsy/head trauma, high fasting blood glucose, a large extent of tumor lesion, and a low degree of tumor differentiation were risk factors for epileptic seizures (OR=1.257-3.823,P<0.05). Conclusion The history of drinking/epilepsy/head trauma, high fasting blood glucose, a large extent of tumor lesion, and a low degree of tumor differentiation are risk factors for the initial symptom of epileptic seizures in patients with glioma. Patients with such conditions should be taken seriously during clinical diagnosis and treatment, and active prevention and treatment should be performed according to the medical advice.

Objective To investigate the differences in vaginal microbiota between women of childbearing age stratified by the age of 35 years, and to provide guidance for the reproductive health of women. Methods A total of 2 683 women of childbearing age who underwent routine physical examination and vaginal microbiota assessment in Center for Reproductive Medicine, Shandong University, from January 2016 to December 2018 were enrolled as subjects, and according to the age, they were all divided <35 years group (group A) and 35-49 years group (group B). The vaginal microbiota were compared between the two groups. Results There were significant differences between groups A and B in the proportion of women with balanced vaginal microbiota, bacterial vaginosis (BV), abnormal dominant bacteria, abnormal pH value, abnormal cleanliness, abnormal sialidase, abnormal Nugent scores, or leukocyte esterase abnormalities, as well as microbial diversity and density (Z=3.94,χ²=5.70-29.43,P<0.05). There were no significant differences between the two groups in the proportion of women with abnormal H₂O₂, N-acetylglucosaminidase, or proline aminopeptidase (P>0.05). Conclusion Compared with the women of childbearing age younger than 35 years, those aged ≥35 years have a higher incidence rate of vaginal microbiota dysbiosis, mainly characterized by an increase in the prevalence rate of BV, impaired vaginal acidic environment, and an increase in the proportion of women with abnormal dominant bacteria.

Objective To investigate the effect of preoperative blood lipid levels on the therapeutic efficacy of sigmoid sinus bone wall reconstruction in patients with pulsatile tinnitus caused by sigmoid sinus disease. Methods A total of 43 patients with pulsatile tinnitus caused by sigmoid sinus disease who underwent sigmoid sinus bone wall reconstruction in Department of Otorhinolaryngology Head and Neck Surgery in our hospital from May 2019 to September 2022 were enrolled in this study, and accor-ding to the degree of improvement in tinnitus after surgery, they were divided into cured group, response group, and non-response group. Related data were collected and compared between groups, including general information, the serum levels of low-density lipoprotein (LDL), high-density lipoprotein, apolipoprotein A1 (APOA1), apolipoprotein B (APOB), free fatty acid (FFA), triglyceride, and total cholesterol (TC), and APOB/APOA1 ratio before surgery. Results Among the 43 patients, 24 were cured, 14 showed response, and 5 showed no response. The serum levels of LDL, APOB, FFA, and TC and APOB/APOA1 ratio were significantly higher than the normal reference values, while the level of APOA1 was significantly lower than the normal refe-rence value (t=3.683-9.713,P<0.05), and the non-response group had a significantly higher APOB/APOA1 ratio than the ot-her two groups (F=7.527,q=3.120,3.650,P<0.05). Conclusion Sigmoid sinus bone wall reconstruction can effectively alleviate the symptoms of patients with pulsatile tinnitus caused by sigmoid sinus disease, and abnormal blood lipid levels before surgery may compromise the efficacy of surgery by interfering with bone reconstruction. Preoperative evaluation of blood lipid levels, especially APOB/APOA1 ratio, can help to predict surgical outcome, thereby providing efficient treatment for patients.

Objective To investigate the value of prethrombotic state molecules and the serum level of complement C1q tumor necrosis factor-related protein 1 (CTRP1) in predicting the prognosis of patients after percutaneous coronary intervention (PCI). Methods A total of 86 patients who underwent PCI in our hospital from January 2018 to February 2020 were enrolled, and according to the presence or absence of major adverse cardiovascular events within 1 year after surgery, they were divided into good prognosis group with 71 patients and poor prognosis group with 15 patients. The two groups were compared in terms of the serum level of CTRP1 and the plasma levels of D-dimer (D-D), thrombus precursor protein (TpP), fibrinogen (Fbg), and von Willebrand factor (VWF) on admission. The receiver operating characteristic (ROC) curve was plotted and the area under the ROC curve (AUC) was calculated to assess the performance of the above indicators in predicting the prognosis of patients after PCI. Results Compared with the poor prognosis group, the good prognosis group had significantly lower plasma levels of D-D, TpP, Fbg, and vWF (t=2.094-5.664,P<0.05) and a significantly higher serum level of CTRP1 on admission (t=2.331,P<0.05). The ROC curve analysis showed that the combination of these indicators had the best performance in assessing the prognosis of patients, with an AUC of 0.935, a sensitivity of 93.33%, and a specificity of 78.87%. Conclusion Prethrombotic state molecules and serum CTRP1 level have a high accuracy in predicting the prognosis of patients after PCI, and these indicators can be monitored in clinical practice to improve the long-term prognosis of patients with coronary heart disease.

Objective To investigate the influence of ADAM metallopeptidase with thrombospondin type 1 motif 9 (ADAMTS9) on the prognosis of patients with thyroid cancer (THCA) and its potential mechanism of action. Methods TCGA database was used to obtain the data on the expression level of ADAMTS9 in THCA tissue and the clinical data of patients, and HPA database was used to obtain the data on the protein expression level of ADAMTS9 in THCA tissue. The expression level of ADAMTS9 was compared between normal tissue and THCA tissue, as well as the degree and intensity of ADAMTS9 protein staining. A functional enrichment analysis was used to predict the biological processes involving ADAMTS9 in THCA. The Kaplan-Meier Plotter database was used to analyze the influence of the expression level of ADAMTS9 on the overall survival time of THCA patients, and gene set enrichment analyses were used to explore the potential signaling pathways associated with ADAMTS9 and important prognostic factors in THCA. Results Compared with normal tissue, THCA tissue showed a significantly higher expression level of ADAMTS9 (t=12.35,P<0.05) and significantly higher degree and intensity of ADAMTS9 protein staining (t=18.68,12.35,P<0.05). The functional enrichment analysis showed that ADAMTS9 mainly participated in the regulation of RNA slicing and gene expression. The patients with high ADAMTS9 expression had a significantly longer overall survival time than those with low expression (HR=0.22,95%CI=0.06-0.76,P<0.05). The pathway analysis showed that the expression level of ADAMTS9 was positively correlated with calcium signaling pathway score (NES=1.852,P<0.05) and was negatively correlated with glycosphingolipid biosynthesis pathway score and citrate cycle pathway score (NES=-1.581,-1.863,P<0.05). Age (HR=1.11,95%CI=1.05-1.67,P<0.05) was an independent prognostic factor for the overall survival time of patients, and significant enrichment of energy metabolism-related pathways was observed in elderly THCA patients, such as citrate cycle and fatty acid metabolism. Conclusion High mRNA and protein expression levels of ADAMTS9 are observed in THCA tissue, which is favorable to the prognosis of patients, possibly by inhibiting energy-related pathways. In addition, the effect of ADAMTS9 might offset the adverse effect of age on prognosis to a certain degree.