Objective To investigate the effects of cinnamic aldehyde(CA)on Bcl-2-associated X protein(Bax)and Bcl-2 homologous antagonist/killer(Bak)in vascular endothelial cells of diabetic ulcer wound tissues,as well as on cell apoptosis.Methods ① Forty-eight healthy SPF-grade male SD rats(5 weeks old,weighing 180～220 g)were randomly assigned to a control group(12 rats)and a diabetes group(36 rats).The diabetic model was established with an intraperitoneal injection of 50 mg/kg STZ-citrate sodium solution and high-fat diet feeding.The diabetes group was further randomly divided into Model group,CA group,and the rb-bFGF group,with 12 animals in each group.Wounds in the Con and Model groups were disinfected and topically treated with normal saline,CA group received topical application of 4 μmol/L CA in PEG 400 gel,and those of the rb-bFGF group were treated with bevacizumab gel.The wound healing rate of each group was calculated at 3,7 and 14 d after intervention.At 14 d after intervention,pathological changes in the wounds were observed with HE staining,and the expression levels of Bax and Bak were detected by Western blotting.② Human umbilical vein endothelial cell line EA.hy926 was treated with 175 mmol/L glucose for 48 h to establish a cell model of high glucose injury.The experimental cells were divided into control group,model group and CA treatment group.Cell scratch test and tube formation test were performed respectively to determine the migration ability and angiogenesis of the cells.The expression levels of Bax and Bak was detected with immunofluorescence assay,and cell apoptosis was detected by TUNEL staining.Results ①The diabetic rats in the Model group exhibited significantly higher blood glucose level(P<0.05),declined wound healing rate at 7 and 14 d after intervention(P<0.05),and enhanced expression levels of Bax and Bak(P<0.05)when compared with the control group.Pathological observation revealed that,at 14 d after intervention,accompanied with inflammatory reactions,dense infiltration of inflammatory cells,fewer new blood vessels,and continuous fluid exudation in the wound were observed in the Model group,but the control group presented complete epithelialization in full-thickness skin.Compared with the conditions in the Model group,both CA and rb-bFGF treatment improved the epithelialization process,with mature granulation tissues,showing good healing condition,promoted wound healing rate(P<0.05),and decreased the expression levels of Bax and Bak(P<0.05).② The results of cell experiments showed that the cells of the model group showed significantly reduced migration ability and tube formation ability(P<0.05),reduced protein levels of Bax and Bak(P<0.05),and lower apoptotic rate(P<0.05)when compared with the cells in the model group.Conclusion CA can inhibit the expression of apoptosis-related proteins Bax and Bak,promote the migration and tube formation of vascular endothelial cells,and inhibit the cell apoptosis under high glucose condition,which may be an important reason for its promoting wound healing in diabetic ulcer rats.

Objective:To construct a novel clinical prognosis signature for hepatocellular carcinoma (HCC) patients using T cell-related ubiquitination genes.Methods:Transcriptome and clinical data of 371 liver cancer and 50 normal samples were obtained from the TCGA database, and microarray sequencing data of 221 liver cancer samples were selected from the GSE14520 dataset. Single-cell RNA sequencing (scRNA-seq) data of HCC patients were analyzed to identify T cell-related marker genes. These were combined with ubiquitination-related genes. Weighted gene co-expression network analysis (WGCNA) was performed on TCGA transcriptome data to select key genes, resulting in the identification of T cell-related ubiquitination genes. A prognostic model was then constructed using LASSO-Cox regression. Finally, a nomogram was created by combining risk scores and clinical parameters. Count data were expressed by examples and percentages(%). Spearman correlation test was used for correlation analysis. Kaplan-Meier method and Log-rank test were used for survival analysis.Results:Initially, 1 458 T cell-related marker genes were identified. Intersection with 797 ubiquitination-related genes led to the identification of 94 common genes. WGCNA analysis revealed the prognosis-related module MEturquoise. After performing differential gene analysis, Kaplan-Meier analysis, and COX regression, 16 candidate genes were confirmed. LASSO-COX algorithm accurately selected five key genes- UBE2S, PSMD1, FBXL5, UBE2E1, and PSMA7—to construct the prognostic model. Kaplan-Meier analysis indicated that the risk score of the prognostic model was significantly associated with the prognosis of HCC patients (Log-rank test=0.001). Both univariate and multivariate COX regression analysed demonstrated that the risk score was an independent prognostic factor for HCC patients( P<0.05). Finally, a nomogram was constructed by combining the risk score and clinical parameters, providing a more accurate prediction of patient prognosis. Conclusion:The T cell-related ubiquitination gene prognostic model can effectively predict the prognosis of patients with liver cancer.

Objective To discuss the application value of CT and DSA multimodal image fusion technology in endovascular interventional therapy for arterial occlusive lesions of lower extremities and to evaluate its efficacy and safety so as to provide a scientific basis for clinical decision-making.Methods A total of 283 lower limbs with arterial complete occlusive lesions,who received treatment at Affiliated Baiyun Hospital of Guizhou Medical University hospital from January 2020 to December 2023,were selected for this study.The 283 diseased lower limbs were randomly divided into study group(n=142)and control group(n=141).In the study group the endovascular interventional therapy assisted by CT and DSA multimodal image fusion technology was adopted,while in the control group the traditional DSA-guided endovascular interventional therapy was employed.The imaging parameters,surgical success rates,X-ray exposure doses,time spent for operation,incidence of postoperative complications,changes of ankle-brachial index(ABI),primary patency rate,assisted primary patency rate,and secondary patency rate were compared between the two groups.Results The surgical success rate in the study group was 96.47％,which was significantly higher than 87.94％in the control group(P＜0.05).The mean time spent for operation in the study group was(125.42±23.74)minutes,which was shorter than(147.81±29.33)minutes in the control group.The mean X-ray exposure dose in the study group was(2 856.34±427.82)mGy·cm2,which was lower than(3 674.53±512.60)mGy·cm2 in the control group.The incidence of postoperative complications in the study group was 4.23％,which was significantly lower than 12.57％in the control group(P＜0.05).The ABI values of the affected limbs in the study group and control group increased from preoperative(0.65±0.15)and(0.60±0.18)respectively to postoperative(1.09±0.32)and(0.90±0.28)respectively.The postoperative ABI value in the study group was higher than that in the control group(P＜0.05).The postoperative 12-month primary patency rate,assisted primary patency rate and secondary patency rate in the study group were 78.17％,85.92％and 90.14％respectively,which were better than 67.38％,75.89％and 80.85％respectively in the control group.Conclusion For arterial occlusive lesions of lower extremities,endovascular interventional therapy with the help of CT and DSA multimodal image fusion technology has high surgical success rates,low incidence of complications,and satisfactory revascularization rate.This technology provides new idea and method for the treatment of arterial occlusive lesions of lower extremities with high clinical safety.Therefore,this technology is worthy of clinical promotion and application.

Objective To discuss the effect of PI3K-AKT signaling pathway regulated by microRNA-155(miRNA-155)targeted protein tyrosine phosphatase non-receptor type 21(PTPN21)on the proliferation,migration and invasion of hepatocellular carcinoma(HCC)cells.Methods Lentivirus transfection was used to silence the expression of miRNA-155 in human Huh7 HCC cells,and real-time fluorescent quantitative polymerase chain reaction(RT-qPCR)was used to detect the silencing effect of miR-155.After obtaining stable cell lines,the cell lines were randomly divided into Blank group(normal Huh7 cells),shNC group(Huh7 cells+empty miR-155 vector),sh-miR-155(Huh7 cells+miR-155 silencing),sh-miR-155+Recilisib group(Huh7 cells+miR-155 silencing+PI3K-AKT agonist),shNC+Recilisib group(Huh7 cells+empty miR-155 vector+PI3K-AKT agonist).Dual luciferase assay was used to determine whether PTPN21 was the downstream of miR-155.The cell proliferation ability of cells in each group was detected by MTT assay.The apoptosis level of each group was tested by flow cytometry.The invasion and migration ability of cells was assessed by Transwell assay.Western blot analysis was used to observe the differences in protein expression of PTPN21,PI3K,P-PI3K,AKT,P-AKT,and apoptosis-related proteins including BAX,BCL-2 and caspase-3 in all groups.Results The expression level of miR-155 in sh-miR-155 group was lower than that in Blank group and shNC group(P＜0.000 1),and the difference in miR-155 expression level between Blank group and shNC group was not statistically significant(P＞0.05).MTT results showed that A values of Huh7 cells at 2,3,4 and 5 day in sh-miR-155 group were lower than those in Blank group and shNC group(P＜0.000 1),while these differences between Blank group and shNC group were not statistically significant(P＞0.05).In sh-miR-155 group the A values at 2,3,4 and 5 day were lower than those in sh-miR-155+Recilisib group and shNC+Recilisib group(P=0.0052 and P＜0.0001,respectively),while the A values at 2,3,4 and 5 day in sh-miR-155+Recilisib were lower than those in shNC+Recilisib group(P＜0.000 1).There was no significant differences in cell migration and number of invasion cells between the Blank group and shNC group(P＞0.05).After activation of PI3K-AKT signaling pathway,the migration and invasion capacity of HCC cells in the shNC+Recilisib group were significantly enhanced when compared with the Blank group(P＜0.000 1).In contrast,the number of migrated and invaded Huh7 cells after miR-155 silencing was significantly lower than that in the Blank group and shNC group(P＜0.000 1)and this phenomenon became reversed by PI3K agonist.Compared with the sh-miR-155 group,in the sh-miR-155+Recilisib group the migration and invasion ability of HCC cells was enhanced(P=0.000 2).Lentiviral transfection of Huh7 human HCC cells to silence miR-155 and downregulate miR-155 inhibiting PTPN21 regulation of the PI3K-AKT signaling pathway,thus inhibiting the invasion,migration and proliferation ability of HCC cells and promoting the apoptosis of HCC cells.Conclusion miR-155 inhibits the migration,invasion and proliferation of HCC cells through targeting PTPN21 regulation of PI3K-AKT signaling pathway.The miR-155 may be a potential therapeutic target for HCC in the future.(J Intervent Radiol,2024,32:44-51)

Objective:To predict the status of microvascular invasion (MVI) in intrahepatic cholangiocarcinoma (ICC) patients preoperatively based on the radiomics analysis of contrast-enhanced CT to provide imaging evidence for early identification of patients at high risk of recurrence.Methods:Clinical data of 40 ICC patients who underwent radical hepatectomy at Nanjing Drum Tower Hospital from January 2021 to May 2024 were retrospectively collected. Patients were divided into the MVI group ( n=8) and the non-MVI group ( n=32) according to the MVI status of the postoperative pathology report. Whether there were differences in each pathological index between the groups and the efficacy of radiomics analysis of contrast-enhanced CT for the preoperative prediction of MVI were analyzed. The regions of interest (ROI) were outlined on the arterial and venous phase images using the 3D Slicer software. Then, radiomics features were extracted from each ROI based on Python. Finally, the LASSO regression and glm function were used to screen radiomics features and establish a prediction model based on the R language. The established predictive model′s diagnostic efficacy, calibration, and net clinical benefit were evaluated using the receiver operating characteristic (ROC) curve, calibration curve, and decision curve analysis (DCA), respectively. Normally distributed measurement data were expressed as mean±standard deviation ( ± s) and compared using the t-test. Count data were expressed as frequency and compared using the chi-square test. Results:Patients in the MVI group had more poorly differentiated tumors and a significantly higher proportion of lymph node metastases ( P<0.05). The established radiomics prediction model included six features, 1 first-order statistical feature and 5 gray texture features. The area under the ROC curve was 0.87, the sensitivity was 75.0%, and the specificity was 90.6%. The calibration curve showed good agreement between the predicted MVI and actual MVI status, and the decision curve demonstrated that the model could provide a large net clinical benefit. Conclusion:Radiomics analysis of contrast-enhanced CT can identify the MVI status of ICC patients preoperatively and aid in clinical decision-making, providing vital evidence for individualized and precise treatment of ICC.

Objective:To explore the risk factors of acute kidney injury(AKI)after liver transplantation(LT), examine its prognostic impact and construct a clinical prediction model.Methods:Clinical data are retrospectively reviewed for 220 LT recipients.They are divided into two groups of AKI(93 cases)and non-AKI(127 cases)according to the occurrence of AKI post-LT.Clinical data of two groups are compared.The variables with statistically significant inter-group differences in univariate analysis are included for multivariate analysis for obtaining the independent risk factors for AKI post-LT.Then the independent risk factors are employed for fitting a prediction model and a visual nomogram is constructed.At the same time, discrimination and calibration of the prediction model are evaluated.Extubation time, length of intensive care unit(ICU)stay, continuous renal replacement therapy(CRRT)rate, length of hospital stay, in-hospital mortality, estimated glomerular filtration rate(eGFR)at discharge, incidence of chronic renal failure(CRF)and readmission times are compared between two groups.Survival analysis is also performed between AKI and non-AKI groups and AKI 0/1 and AKI 2/3 stages.Results:The incidence of AKI post-LT is 42.3%.Age( OR=1.036, 95% CI: 1.001~1.073), preoperative serum creatinine level( OR=1.030, 95% CI: 1.011~1.049), platelet count( OR=0.992, 95% CI: 0.985~0.999), Child-Pugh class C( OR=2.678, 95% CI: 1.031~6.952), postoperative abdominal infection( OR=2.271, 95% CI: 1.120~4.603)and abdominal hemorrhage( OR=3.869, 95% CI: 1.016~14.72)are independent risk factors for AKI post-LT.The AUC/C-index of nomogram prediction model is 0.789 with a Brier score of 0.183, showing decent discrimination and calibration.According to the nomogram score, the recipients with a risk of AKI>50% are included into high-risk group while those with a risk of AKI<50% into low-risk group.Postoperative survival of low-risk group is better than that of high-risk group( P<0.001).Compared with non-AKI group, AKI group had a later extubation time( P=0.003), a longer length of ICU stay( P<0.001)and hospital stay( P=0.001), a higher rate of CRRT usage( P<0.001)and in-hospital mortality( P<0.001), a lower eGFR at discharge( P<0.001)and a higher incidence of CRF( P<0.001).Postoperative survival of non-AKI group was better than that of AKI group( P=0.048).Postoperative survival of patients with AKI 0/1 is better than that of those with AKI 2/3( P=0.002). Conclusions:Advanced age, high preoperative serum creatinine, low preoperative platelet, poor preoperative liver function, postoperative abdominal infection and abdominal hemorrhage may elevate the risks of AKI post-LT.And the nomogram prediction model based upon the above risk factors has a high value of clinical application.

Objective:To investigate the efficacy and safety of bladder neck resection combined with multipoint injection of triamcinolone acetonide in the treatment of bladder neck contractures (BNC) after transurethral resection prostate (TURP).Methods:The data of 25 patients with BNC after TUPR who underwent transurethral surgery in West China Hospital of Sichuan University from July 2019 to November 2021 were retrospectively analyzed, and the patients were divided into 2 groups according to the treatment method. There were 15 cases in the steroid injection group, with an average age of (67.5±8.8) years, 1 case of diabetes, and 2 cases of hypertension. And the average postoperative time after TURP was (21.9±29.1) months, the preoperative International Prostate Symptom Score (IPSS) was (30.0±3.5) points, quality of life (QOL) score was (5.7±0.5) points. There were 10 cases in the non-steroid injection group, with an average age of (65.2±10.5) years, 1 case of diabetes, and 2 cases of hypertension. And the average postoperative time of TURP was (29.3±33.5) months, and the preoperative IPSS was (30.4±2.6) points, QOL score was (5.8±0.4) points. There was no significant difference between the two groups ( P>0.05). In the steroid injection group, bladder neck resection combined with multipoint injection of triamcinolone acetonide was performed. The patients were in the lithotomy position, and a scope was placed through the urethra into the distal end of the narrow urethra under direct vision for observation, and a supersmooth guide wire was placed. The narrow section is then incised at 6 o'clock. After replacing the resectoscope, the urethral stricture scar was excised until the normal tissue of the bladder neck was exposed. Hemostasis by electrocoagulation resulted in no active bleeding from the urethra after resection. After the bladder injection needle was inserted, 80 mg (12 ml) of triamcinolone acetonide injection was injected in 6 equally spaced needles at the direction of the bladder neck from 3 to 9 o'clock. A supersmooth guide wire was placed, and a three-channel silicone urinary catheter was indwelled along the guide wire. The non-steroid injection group underwent simple bladder neck resection. Re-examination of urethroscopy at 3 and 6 months after operation showed that the patient had obvious dysuria and the microscopic examination showed that the recurrence of bladder neck stenosis was defined as the recurrence of stenosis. The efficacy, complications, and recurrence-free survival rates of the two groups were compared. Predictors of postoperative BNC recurrence were analyzed. Results:The operations in both groups were successfully completed. The operation time of the steroid injection group and the non-steroid injection group were (36.0±17.8) min and (48.5±57.9) min respectively ( P=0.438), and the intraoperative blood loss was (1.9±3.0) ml and (12.0±31.1) ml ( P=0.221)respectively. The length of hospital stay was (5.8±1.2) d and (4.4±2.5) d, respectively ( P=0.070). There was 1 case of transient hematuria and 1 case of epididymitis in the steroid injection group, and 1 case of transient hematuria and 1 case of transient dysuria in the non-steroid injection group ( P>0.05), all of which were relieved after symptomatic treatment. The IPSS of steroid injection group and non-steroid injection group were (11.0±5.6) points and (12.4±3.9) points at 3 months after operation, and (10.1±4.9) points and (14.7±7.7) points at 6 months after operation, respectively. QOL at 3 months after operation was (1.7±1.2) points and (2.1±1.5) points, respectively, and at 6 months after operation, it was (1.5±1.3) points and (3.0±2.0) points, respectively. There was statistical significance ( P<0.05). There were 1 case and 2 cases of recurrence in the steroid injection group and non-steroid injection group at 3 months after operation ( P=0.543), and 1 case and 5 cases at 6 months after operation ( P=0.023). The difference in survival curve between the two groups was statistically significant ( P=0.013). Combined steroid therapy ( OR=14.000, 95% CI1.299-150.889, P=0.030), time after scar resection ( OR=1.138, 95% CI1.017-1.273, P=0.025), postoperative IPSS( OR=1.302, 95% CI1.018-1.666, P=0.036), postoperative QOL score ( OR=4.280, 95% CI1.523-12.030, P=0.006) were the predictors of stenosis recurrence 6 months after surgery. Conclusion:Bladder neck scar resection combined with steroid injection could be safe and effective in the treatment of BNC after TURP, and local steroid injection may help reduce the recurrence rate of postoperative stenosis.

Objective:To explore the efficient construction of HSF1 gene knockout mouse model using CRISPR/Cas9 gene editing technology, and to establish the early basis for the mouse model of primary osteosarcoma.Methods:According to exon 9 of HSF1 gene structure, the corresponding GRNA (guideRNA) was selected and screened. Then the transcription template of sgRNA (small guide RNA) was amplified by PCR, and four up stream primers were obtained. Subsequently, sgRNA was transcribed in vitro and screened by Tube Screen platform to screen the sgRNA with effective cutting, and the sgRNA with the highest cutting efficiency was selected from the screening results for subsequent experiments. The transcription template of SPCas9mRNA was amplified by PCR, and then Cas9mRNA was transcribed in vitro. The sgRNA transcribed in vitro and Cas9mRNA were injected into the fertilized eggs of healthy C57BL/6 mice, and the tissue was extracted from the tail of the born mice and identified by PCR sequencing. Heterozygous female mice of F0 generation were selected to mate with wild-type male mice too btain F1 generation off spring. The mutation of gene bases of F1 generation mice was detected by AGAR gel electrophoresis and gene sequencing. The heterozygous male mice of the F1 generation and female mice of the F0 generation were back crossed to obtain the F2 generation daughter mice. The tail tissues were cut and sequenced to obtain the F2 generation homozygous knockout mice. PCR was used to observe the cutting efficiency of sgRNA and the sequencing of rat tail tissue, and SNAPGene software was used for gene sequence alignment to determine the deletion of base fragments.Results:The up stream primers sgRNA-1 Primer-f, sgRNA-2 Primer-f, sgRNA-3 Primer-f, sgRNA-4 Primer-f and down stream primers sgRNA-4 Primer -r were obtained by PCR amplification. After in vitro tran scription and screening of sgrRNA, sgrRNA-1, sgrRNA-2 and sgrRNA-4 had high cleavage efficiency and were selected for subsequent experiments. T7 promoter was added to the 5 'end of Cas9 mRNA, and Cas9 mRNA was obtained by PCR and in vitro transcription kit. Mixed Cas9-sgRNA solution was injected into the fertilized eggs of mice and cultured. The cultured two-cell fertilized eggs were injected into the ampulla of the pseudo pregnant female mice, and the F0 generation mice were obtained successfully. A total of 8 heterozygous mice of F0 generation were obtained by Agar gel electrophoresis. Three heterozygous knockout mice of F1 generation were obtained by breeding the female heterozygous mice of F0 generation with healthy wild-type male mice and PCR and sequencing. Three heterozygous male mice of F1 generation were back crossed with female mice of F0 generation 3 to obtain F2 generation mice. Through the observation of electrophoresis and sequencing results of F2 generation mice, it was confirmed that 7 mice were missing HSF1 base sequence, and the electrophoresis results showed mutant bands and no wild-type bands, which were identified as homozygous. The F2 generation homozygous mice were able to breed stably. As eries of results proved that the HSF1 gene knockout mouse model was successfully established in this experiment.Conclusion:CRISPR/Cas9 technology was successfully used to construct HSF1 gene knockout mouse model, with strong stability and high reproducibility, which laida foundation for further study of HSF1 gene expression products and establishment of mouse model of primary osteosarcoma.

Objective:To compare ultrasound (US) guided versus computed tomography (CT) guided radiofrequency ablation (RFA) in treatment of early hepatocellular carcinoma (HCC).Methods:The data of 133 patients with early HCC treated by RFA in the Department of Hepatobiliary Surgery of Shandong Provincial Hospital from February 1, 2015, to January 31, 2017, was analyzed retrospectively. These patients were divided into two groups: the US-guided group and the CT-guided group. The clinical data was collected and the factors affecting prognosis were analyzed.Results:Compared with the CT-guided group, the operation time of the US-guided group was significantly shorter [(29.0±12.0)min vs. (55.0±19.0)min, P<0.05], but the number of ablation sessions per tumor was significantly less [(1.1±0.3) vs. (2.0±0.6), P<0.05]. There was no significant difference in the complete ablation rates, postoperative complication rates and postoperative length of hospital stay between the two groups ( P>0.05). The CT-guided group was superior to the US-guided group in the local tumor recurrence and progression-free survival rates ( P<0.05). On multivariate analysis, CT-guided RFA was an independent protective factor for local tumor recurrence ( HR=0.266, 95% CI: 0.073-0.967, P<0.05) and progression-free survival ( HR=0.415. 95% CI: 0.213-0.806, P<0.05), while AFP >20 ng/ml ( HR=4.821, 95% CI: 1.714-13.560, P<0.05) was an independent risk factor for progression-free survival. Conclusion:CT-guided percutaneous RFA was superior to US-guided RFA in local treatment of early HCC, probably related to more needle placements and longer ablation time under CT guidance.