AIM: To investigate the role of pyroptosis in the development of diabetic retinopathy(DR)and to explore the regulatory mechanism by which circular RNA(circRNA)and its targeted microRNA(miRNA)mediate pyroptosis in DR, providing new therapeutic targets and strategies for the prevention and treatment of DR.METHODS: A streptozotocin(STZ)-induced model of type 1 diabetes in SD rats was established. The expression of circRNA_0076631, miR-214, and pyroptosis-related factors were measured in retinal tissues. CCK-8 and tube formation assays were used to detect the effect of different concentration of glucose on cell proliferation and angiogenic abilities of human retinal microvascular endothelial cells(HRMECs). The expression levels of circRNA_0076631, miR-214, and pyroptosis-related markers were evaluated through qRT-PCR and Western blot analysis, with additional experiments conducted following circRNA_0076631 knockdown to assess its effect on pyroptosis markers. Previous bioinformatics analysis and luciferase reporter assays identified a shared binding site among circRNA_0076631, miR-214, and caspase-1. To clarify the interaction between these molecules, co-transfection experiments using circRNA_0076631 inhibitors(ASO-circRNA_0076631), miR-214 overexpression transfection reagent, and miR-214 inhibitors(AMO-miR-214)were conducted to elucidate the regulatory pathway involved in DR.RESULTS: Both the diabetic rat model and D-glucose-treated HRMECs showed significantly elevated expression of circRNA_0076631 and pyroptosis-related factors(NLRP3, caspase-1, and IL-1β), while miR-214 expression was reduced(all P<0.05). The mRNA expression of pyroptosis-related factors caspase-1 was reduced after the overexpression of miR-214, and it was upregulated after the inhibition of miR-214(all P<0.05). Knockdown of circRNA_0076631 reduced the mRNA expression of pyroptosis markers caspase-1(P<0.05). Co-transfection experiments revealed that the inhibition circRNA_0076631 suppressed pyroptosis(all P<0.05), but this suppression was reversed upon co-transfection with miR-214 inhibitors, leading to increased mRNA expression of the pyroptosis marker caspase-1(all P<0.05).CONCLUSION: The circRNA_0076631 and pyroptosis play critical roles in the pathogenesis of DR, and circRNA_0076631 may regulate pyroptosis by modulating miR-214, which in turn influences the expression of caspase-1 in the pyroptosis signaling pathway, thereby contributing to DR progression. The circRNA_0076631 may serve as a novel therapeutic target, providing new insights for the prevention and treatment of DR.

AIM: To investigate the role of pyroptosis in the development of diabetic retinopathy(DR)and to explore the regulatory mechanism by which circular RNA(circRNA)and its targeted microRNA(miRNA)mediate pyroptosis in DR, providing new therapeutic targets and strategies for the prevention and treatment of DR.METHODS: A streptozotocin(STZ)-induced model of type 1 diabetes in SD rats was established. The expression of circRNA_0076631, miR-214, and pyroptosis-related factors were measured in retinal tissues. CCK-8 and tube formation assays were used to detect the effect of different concentration of glucose on cell proliferation and angiogenic abilities of human retinal microvascular endothelial cells(HRMECs). The expression levels of circRNA_0076631, miR-214, and pyroptosis-related markers were evaluated through qRT-PCR and Western blot analysis, with additional experiments conducted following circRNA_0076631 knockdown to assess its effect on pyroptosis markers. Previous bioinformatics analysis and luciferase reporter assays identified a shared binding site among circRNA_0076631, miR-214, and caspase-1. To clarify the interaction between these molecules, co-transfection experiments using circRNA_0076631 inhibitors(ASO-circRNA_0076631), miR-214 overexpression transfection reagent, and miR-214 inhibitors(AMO-miR-214)were conducted to elucidate the regulatory pathway involved in DR.RESULTS: Both the diabetic rat model and D-glucose-treated HRMECs showed significantly elevated expression of circRNA_0076631 and pyroptosis-related factors(NLRP3, caspase-1, and IL-1β), while miR-214 expression was reduced(all P<0.05). The mRNA expression of pyroptosis-related factors caspase-1 was reduced after the overexpression of miR-214, and it was upregulated after the inhibition of miR-214(all P<0.05). Knockdown of circRNA_0076631 reduced the mRNA expression of pyroptosis markers caspase-1(P<0.05). Co-transfection experiments revealed that the inhibition circRNA_0076631 suppressed pyroptosis(all P<0.05), but this suppression was reversed upon co-transfection with miR-214 inhibitors, leading to increased mRNA expression of the pyroptosis marker caspase-1(all P<0.05).CONCLUSION: The circRNA_0076631 and pyroptosis play critical roles in the pathogenesis of DR, and circRNA_0076631 may regulate pyroptosis by modulating miR-214, which in turn influences the expression of caspase-1 in the pyroptosis signaling pathway, thereby contributing to DR progression. The circRNA_0076631 may serve as a novel therapeutic target, providing new insights for the prevention and treatment of DR.

Objective To analyze the application value of transvaginal ultrasonography(TVS)combined with contrast-enhanced ultrasound(CEUS)in the diagnosis of cesarean scar pregnancy(CSP).Methods Sixty patients with CSP confirmed by operation and pathology admitted to the First Hospital of Jiaxing from January 2022 to November 2023 were retrospectively analyzed.TVS and CEUS were examined before operation.The detection rate and type diagnosis of CSP were compared between TVS and CEUS alone and combined diagnosis.The receiver operating characteristic(ROC)curve was used to analyze the efficiency of TVS and CEUS in identifying gestational cesarean scar pregnancy(GCSP).Results Among the 60 patients with CSP,53 patients with GCSP and 7 patients with mass cesarean scar pregnancy(MCSP)were confirmed by surgery and pathology.The detection rate of CSP by TVS combined with CEUS(98.33％)was significantly higher than that by TVS or CEUS(P＜0.05),and the detection rate of CSP by CEUS was significantly higher than that by TVS(P＜0.05).The detection rate of GCSP by TVS combined with CEUS(100.00％)was significantly higher than that by TVS or CEUS(P＜0.05).The area under the curve(AUC)of TVS combined with CEUS for identifying GCSP was the largest,which was 0.857.The sensitivity was 100.00％,and the specificity was 71.40％.The detection rate of MCSP by TVS combined with CEUS was significantly higher than that by TVS(P＜0.05).Conclusion TVS combined with CEUS can effectively improve the clinical detection rate of CSP,which is conducive to the diagnosis of CSP classification,and has important application value.

Purpose: This study aimed to identify prognostic factors for patients with distant lymph node-involved gastric cancer (GC) using a machine learning algorithm, a method that offers considerable advantages and new prospects for high-dimensional biomedical data exploration. Materials and Methods: This study employed 79 features of clinical pathology, laboratory tests, and therapeutic details from 289 GC patients whose distant lymphadenopathy was presented as the first episode of recurrence or metastasis. Outcomes were measured as anycause death events and survival months after distant lymph node metastasis. A prediction model was built based on possible outcome predictors using a random survival forest algorithm and confirmed by 5×5 nested cross-validation. The effects of single variables were interpreted using partial dependence plots. A contour plot was used to visually represent survival prediction based on 2 predictive features. Results: The median survival time of patients with GC with distant nodal metastasis was 9.2 months. The optimal model incorporated the prealbumin level and the prothrombin time (PT), and yielded a prediction error of 0.353. The inclusion of other variables resulted in poorer model performance. Patients with higher serum prealbumin levels or shorter PTs had a significantly better prognosis. The predicted one-year survival rate was stratified and illustrated as a contour plot based on the combined effect the prealbumin level and the PT. Conclusions Machine learning is useful for identifying the important determinants of cancer survival using high-dimensional datasets. The prealbumin level and the PT on distant lymph node metastasis are the 2 most crucial factors in predicting the subsequent survival time of advanced GC.Trial Registration: ChiCTR Identifier: ChiCTR1800019978

Purpose: While several prognostic models for the stratification of death risk have been developed for patients with advanced gastric cancer receiving first-line chemotherapy, they have seldom been tested in the Chinese population. This study investigated the performance of these models and identified the optimal tools for Chinese patients. Materials and Methods: Patients diagnosed with metastatic or recurrent gastric adenocarcinoma who received first-line chemotherapy were eligible for inclusion in the validation cohort. Their clinical data and survival outcomes were retrieved and documented. Time-dependent receiver operating characteristic (ROC) and calibration curves were used to evaluate the predictive ability of the models. Kaplan-Meier curves were plotted for patients in different risk groups divided by 7 published stratification tools. Log-rank tests with pairwise comparisons were used to compare survival differences. Results: The analysis included a total of 346 patients with metastatic or recurrent disease.The median overall survival time was 11.9 months. The patients were different into different risk groups according to the prognostic stratification models, which showed variability in distinguishing mortality risk in these patients. The model proposed by Kim et al. showed relative higher predicting abilities compared to the other models, with the highest χ 2 (25.8) value in log-rank tests across subgroups, and areas under the curve values at 6, 12, and 24 months of 0.65 (95% confidence interval [CI]: 0.59–0.72), 0.60 (0.54–0.65), and 0.63 (0.56–0.69), respectively. Conclusions Among existing prognostic tools, the models constructed by Kim et al., which incorporated performance status score, neutrophil-to-lymphocyte ratio, alkaline phosphatase, albumin, and tumor differentiation, were more effective in stratifying Chinese patients with gastric cancer receiving first-line chemotherapy.

Objective To discuss the clinical characteristics for lateral cervical lymph node metastasis in stage cN0 papillary thyroid microcarcinoma and significance and feasibility of preventive dissection,and provide reference for clinical treatment.Methods Reviewd the clinical data of 191 patients with stage cN0 papillary thyroid microcarcinoma patients from Jul.2011 to Dec.2016 underwent surgery in the Department of General Surgery of Lianyungang Oriental Hospital.Assessed the need for preventive cervical lymph node dissection.Chisquare test and logistic regression were used to analyze the relationship between cervical lymph node metastasis and gender,age,tumor number,tumor size,capsule infiltration,single and bilateral tumors,Hashimoto's disease,and central lymph node metastasis.Results The positive rate of cervical lymph node metastasis in papillary thyroid microcarcinoma was 27.9％ (50/191).Univariate analysis showed that the metastasis of the cervical lymph nodes was associated with infiltration of the capsule,Hashimoto disease,and CLN metastasis (all P ＜ 0.05).Multivariate logistic regression analysis showed that the capsule infiltration (OR =7.563,P =0.000),Hashimoto's disease (OR =4.635,P =0.003),and central lymph node metastasis (OR =3.075,P ＜ 0.001) were able to be independent risk factors for cervical lymph node metastasis.When the positive number of lymph node metastasis in the central region was ≥ 2,the positive rate of cervical lymph nodes was significantly increased (P ＜ 0.001).Eleven patients (5.8％) had temporary recurrent laryngeal nerve palsy,29 patients (15.1％) had transient hypoparathyroidism,and no patients with permanent recurrent laryngeal nerve palsy and hypoparathyroidism.Conclusions The removal of the cervical lymph nodes helps to accurately classify the tumor and assess the risk.It is important to choose the postoperative treatment follow-up plan for patients.For patients with capsule infiltration,Hashimoto's disease,and central lymph node metastasis,cervical lymph node dissection should be routinely performed.

Objective To investigate the effect of neuropilin-1 (NRP1) on radiation-induced epithelial-mesenchymal transition (EMT) by measuring the expressions of EMT-related transcription factors in the irradiated cells with different levels of NRP 1.Methods Human lung type Ⅱ epithelial cells (A549) were transfected with NRP1 over-expression lentiviral vector and NRP1 inhibition vector to construct two cell models of NRP1high-A549 and NRP1low-A549.A NRP1 knock-down cell model was also constructed by transferring siNRP1 into normal mouse lung epithelial MLE-12 cells that was validated at both protein and mRNA levels.A single dose of 10 Gy X-ray was delivered to these cell models,then total protein and RNA were extracted at 0,12,24 and 48 h after irradiation.The expressions of EMT-related transcription factors (Twist and ZEB1) and EMT markers (β3-catenin,N-cadherin,and Vimentin) in each cell model were detected by Western blot and qPCR.Results After 10 Gy irradiation,the expressions of NRP1 mRNA and protein were significantly increased in A549 and MLE-12 cells.The expressions of the mesenchymal markers (Vimentin and N-cadherin) and the transcription factors of ZEB1 and Twist were also significantly increased (A549:t=2.917,7.361,4.852,9.278,P＜0.01;MLE-12:t=9.652,31.357,30.985,17.266,P ＜0.01).The expressions of Vimentin and N-cadherin were significantly decreased in NRP1low-A549 (t =10.077,15.707,P ＜ 0.01) and siNRP1-MLE-12 cells (t =5.745,P ＜ 0.01),but the expression of epithelial marker (β3-catenin) was significantly increased in these cells.The expressions of N-Cadherin and Vimentin were significantly elevated (t =16.055,5.560,P ＜ 0.01),while β-catenin decreased significantly in NRP1high-A549 cells.After irradiation,the transcription factor of Twist in NRP1low-A549 group was significantly decreased (t=3.987,P＜0.01),while the transcription factors of ZEB1 and Twist in the NRP1high-A549 group increased in a time-dependent manner (t =11.289,2.903,P＜0.01).After irradiation,the transcription factor of ZEB1 decreased significantly in siNRP1-MLE-12 cells (t=13.449,P＜0.01),and the protein expressions of ZEB1 and Twist in siNRP1-MLE-12 cells were lower than those of control group in a time-dependent manner.Conclusions NRP1 promotes radiation-induced EMT in human and mouse epithelial cells through up-regulation of transcription factors of ZEB1 and Twist.

Objective To improve the chemotherapy drug delivery to tumor by enhancing the tumor vascular perfusion induced by diagnostic ultrasound combined with microbubbles.Methods Ten healthy male sprague-dawley(SD)rats with total twenty walker-256 tumors implanted in the two back legs were randomized to the two paired groups: controlled group(C,n=10)and treatment group(T,n=10).Tumors in the controlled group were ultrasonic sham operated,while in the treatment group were treated by diagnostic ultrasound combined with microbubbles.The treatment group were taken contrast-enhanced ultrasound(CEUS)before and after treatment and analyzed the quantitative parameters.The microbubbles used in the treatment and CEUS was a kind of self-made lipid microbubbles called Zhifuxian.The 0.02 ml microbubbles were bolus injected at CEUS,while during treatment,0.04 ml microbubbles diluted into 1 ml saline solution were injected slowly at constant speed.Flushed by saline solution after treatment,the rats' tumors were harvested into three parts: one for chemotherapy drug concentration detected by high performance liquid chromatography(HPLC),one for HE detection,and one for Dox fluorescence intensity detected by confocal laser scanning microscopy(CLSM).The peak intensity(PI)values,the area under curve(AUC)values and the Dox concentration of each group were analyzed by pared-samples t test.Results(1)The contrast enhanced ultrasound quantitative analysis of the T group: PI value of the tumors before and after treatment were 66.22±16.25 and 75.74±17.67.The AUC values were 2937.52±677.51 and 3354.91±796.15.There was significant statistical difference between them(t=-5.212,-5.259,all P ＜ 0.05).(2)The Dox concentration of the T and C groups were(1.15±0.25)ug/g and(0.96±0.21)ug/g.There was significant statistical difference between them(t=2.403,P＜0.05).The Dox concentration of the treatment group was 1.2 times of the controlled group.(3)The pathology results of T and C groups: the tumor cells were arranged in cords,with big round deep-stained nucleus.No pathological changes were observed in the controlled group,and there was no significant difference between the two groups.But in the treatment group,tumor vascular congestion and inflammatory cell infiltration could be observed.(4)The confocal laser scanning microscopy(CLSM)detection of the T and C groups: the Dox red fluorescence was distributed in the tumor tissue interstitial,and the fluorescence intensity and distribution area of the treatment group were significant higher than the controlled group.Conclusions Diagnostic ultrasound combined with microbubbles treatment could significantly increase the blood perfusion in the walker-256 tumors of SD rats.Taking advantage of this vascular effect,the chemotherapy drug Dox could be delivered much more to the tumor tissue along with circulating bloodstream.With the addition of the sonoporation effect induced by the cavitation of the microbubbles,the chemotherapy drugs could be released much more to the tumor interstitial tissue.

Objective To investigate the clinical diagnostic value of plasma soluble leukocyte differentiation antigen 14 (presepsin) in patients with rheumatoid arthritis (RA) complicated with pulmonary bacterial infection.Methods A total of 133 patients with RA and 60 healthy controls were enrolled in this study.Fifty-eight RA patients were infected with lung bacterial infection and 75 patients were non-infected with RA.Among them,RA activity was performed in 43 patients and RA was stable in 32 patients.Chemilluminescence immunoassay was used to detect presepsin (P-SEP) in all subjects,and its correlation with inflammatory markers such as white blood cells,blood sedimentation rate and C-reactive protein was analyzed.Results The P-SEP of RA (561 ±142) pg/ml combined with pulmonary bacterial infection group was significantly higher than that of active RA group (378±100) pg/ml (t=8.12,P＜0.01),higher than that of stable RA group (197±68) pg/ml (t=8.51,P＜0.01) and healthy control group (113±9) pg/ml (t=13.75,P＜0.01).There was a positive correlation between P-SEP and leukocyte count in patients with RA complicated with pulmonary bacterial infection (r=0.627,P＜0.01).The degree of the disease activity was correlated with CRP (r=0.63,P＜O.O1),regardless of the P-SEP level (r=0.47,P=0.521).The optimal threshold value of P-SEP in diagnosing RA patients with bacterial infection was 458.9 pg/ml,with a sensitivity of 79.3％ and a specificity of 81.4％.Conclusion P-SEP has an important diagnostic value for the identification of bacterial infection in RA patients,which is not related to RA disease activity.

Objective To explore the effect of low intensity ultrasound (LUS) and microbubble enhanced ultrasound cavitation (MEUS), alone or in combination, on ethanol ablation (EA) of rabbit liver and observe the changes of liver perfusion and liver function. Methods Sixty-two healthy New Zealand rabbits were randomized to five groups: LUS group (n=6), EA group (n=14), LUS + EA group (n=14), MEUS + EA group (n=14), MEUS + LUS + EA group (n=14). For LUS, pulsed low intensity focused ultrasound emission was adopted (focal distance, 15 cm; duty cycle, 0.036%-0.22%;therapy duration, 5-6 min). According to the experiment design, surgically exposed left lobe of the liver was treated. In the LUS, MEUS + EA, and MEUS + LUS + EA groups, quantitative parameters were calculated and compared between the experimental and control liver lobes after different treatments. Three rabbits in each of the EA, LUS + EA, MEUS + EA, and MEUS + LUS + EA groups were used to detect the contents of alanine aminotransferase (ALT) and aspartate transaminase (AST) in arterial blood at five different time points (before treatment, 1 h, 48 h, 5 d, and 7 d after treatment). The livers of the remaining rabbits were harvested for measurement of ethanol ablation volume by drainage method or examination of the histological changes by HE staining 48 h after treatment. Results In the LUS group, the peak intensity (PI) and the area under the curve (AUC) were higher in the experiment lobe than in the control lobe, but there was no significant difference. In the MEUS + EA and MEUS+LUS+EA groups, the PI and AUC values were significantly lower in the experiment lobe than in the control lobe(PI:51.65±16.90 vs 101.09±14.41,44.08±8.46 vs 113.40±9.35;AUC:2183.06±501.13 vs 4258.54±841.21,1900.39±352.59 vs 4385.55±1198.16;t=9.059,16.835,9.630,7.932,P<0.001 for all). In the LUS group, no necrosis was observed, and the necrosis volume was 0 ml. The necrosis volumes caused by ethanol ablation in the EA, LUS+EA, MEUS+EA, and MEUS+LUS+EA groups were (0.84±0.27) ml, (2.42±1.11) ml, (3.52±1.34) ml, and (4.01±1.45) ml. The ethanol ablation volume was significantly lower in the EA group than in the other three groups (u=-13.800, -20.400, -23.400, P<0.05 for all),although there were no significant difference between any two of the latter three groups. No pathological changes were observed in the ultrasound exposed liver of the LUS group. In contrast, a wide range of coagulation necrosis area was noted in the other four groups. Compared with pre-treatment values, ALT and AST levels in all groups showed a slight rise after treatment, peaked at 48 h, and gradually returned to the pretreatment levels after seven days. The tendency of changes in ALT and AST levels with time was similar among the four groups (F=0.256, P=0.855; F=0.517, P=0.686). Conclusion LUS and MEUS, alone or in combination, could significantly increase the ethanol ablated volume of rabbit liver without aggravating liver function.