Objective To observe the value of ultrasound convolutional neural network(CNN)model for assisting distinguishment of cervical lymph node lymphoma and atypical reactive hyperplasia.Methods Totally 335 cases of cervical lymph node lymphoma and 335 cases of atypical reactive hyperplasia were retrospectively enrolled,including 520 cases in development group(260 cases of lymphoma and 260 cases of atypical reactive hyperplasia)and 150 cases in validation group(75 cases of lymphoma and 75 cases of atypical reactive hyperplasia).Patients in development group were divided into training set(182 cases of lymphoma and 182 cases of atypical reactive hyperplasia)and test set(78 cases of lymphoma and 78 cases of atypical reactive hyperplasia)at the ratio of 7∶3.One target lymph node was selected for each case,and one gray-scale ultrasound image and one CDFI were included for training,testing CNN models and verifying the auxiliary efficacy of the models.Based on ultrasound images in training set,5 CNN models,including AlexNet,VGG16,ResNet18,DenseNet161 and EfficientNet-B0,were constructed and trained for distinguishing cervical lymph node lymphoma and atypical reactive hyperplasia,and the models were tested in test set to screen out the best one with the highest classification accuracy.The efficacy of the best CNN model for assisting distinguishment of cervical lymph node lymphoma and atypical reactive hyperplasia was observed.Results Among 5 CNN models,the accuracy of ResNet18 model in test set was the highest(78.21％),and ResNet18 model was regarded as the best CNN model,its sensitivity,specificity and accuracy for assisting distinguishment of cervical lymph node lymphoma and atypical reactive hyperplasia were all higher than those of independent diagnosis made by ultrasound physicians(all P＜0.01).Conclusion The constructed ResNet18 model could be used to effectively assist differentiating cervical lymph node lymphoma and atypical reactive hyperplasia.

Objective To investigate the effect of ginsenoside Re on angiotensin Ⅱ(Ang Ⅱ)-induced proliferation,migration,and phenotype transformation of vascular smooth muscle cells(VSMCs)by regulating the Ras homologous gene family member A(RhoA)/mitogen activated protein kinase(MAPK)pathway.Methods MOVAS cells were divided into control group,Ang Ⅱ group,ginsenoside Re low dose group,ginsenoside Re medium dose group,ginsenoside Re high dose group,andginsenoside Re high dose+RhoA activator(U46619)group.MOVAS cell proliferation was detected by CCK-8.MOVAS cell migration was detected by scratch assay.Immunocytochemistry was used to detect the positive expression of α-smooth muscle actin(α-SMA)and osteopontin(OPN)proteins in MOVAS cells.qRT-PCR was used to detect the mRNA expression of PCNA,MMP-9,and MMP-2 in MOVAS cells.Western blot was used to detect RhoA,phosphorylated extracellular signal regulated kinase 1/2(p-ERK1/2),phosphorylated stress activated protein kinase 1(p-JNK1),and p-P38 protein in MOVAS cells.Results Compared with the control group,the OD450 value,scratch healing rate,OPN protein positive expression,PCNA,MMP-9,MMP-2 mRNA expression,RhoA,p-ERK1/2,p-JNK1,p-P38 protein expression of MOVAS cells in the Ang II group increased,while the positive expression of α-SMA protein decreased significantly(P<0.05).Compared with the Ang Ⅱ group,the OD450 value,scratch healing rate,OPN protein positive expression,PCNA,MMP-9,MMP-2 mRNA expression,and RhoA,p-ERK1/2,p-JNK1,and p-P38 proteins of MOVAS cells in the low,medium,and high dose groups of ginsenoside Re decreased,while the positive expression of α-SMA protein increased.The trend was most significant in the high dose group of ginsenoside Re(P<0.05).Compared with ginsenoside Re high-dose group,OD450 value,scratch healing rate,OPN protein positive expression,PCNA,MMP-9,MMP-2 mRNA expression and RhoA,p-ERK1/2,p-JNK1,p-P38 protein in ginsenoside Re high-dose+U46619 group increased,the positive expression of α-SMA protein decreased significantly(P<0.05).Conclusion Ginsenoside Re may inhibit the proliferation,migration,and phenotype transformation of Ang Ⅱ in MOVAS cells by suppressing the RhoA/MAPK pathway.

Endometrial carcinoma (EC) is one of the most common gynecologic malignancies, with the majority of patients diagnosed at an early stage. Surgery is the primary treatment modality, and postoperative risk stratification plays a key role in guiding adjuvant therapy decisions. Lymphovascular space invasion (LVSI) is an important pathological feature that affects the prognosis of patients and serves as one of the key factors in risk stratification. This review summarizes the prognostic significance of LVSI in patients with early-stage EC and its impact on the selection of adjuvant therapies, aiming to provide new insights for exploring precise and individualized diagnosis and treatment.

Objective To investigate the effect of ginsenoside Re on angiotensin Ⅱ(Ang Ⅱ)-induced proliferation,migration,and phenotype transformation of vascular smooth muscle cells(VSMCs)by regulating the Ras homologous gene family member A(RhoA)/mitogen activated protein kinase(MAPK)pathway.Methods MOVAS cells were divided into control group,Ang Ⅱ group,ginsenoside Re low dose group,ginsenoside Re medium dose group,ginsenoside Re high dose group,andginsenoside Re high dose+RhoA activator(U46619)group.MOVAS cell proliferation was detected by CCK-8.MOVAS cell migration was detected by scratch assay.Immunocytochemistry was used to detect the positive expression of α-smooth muscle actin(α-SMA)and osteopontin(OPN)proteins in MOVAS cells.qRT-PCR was used to detect the mRNA expression of PCNA,MMP-9,and MMP-2 in MOVAS cells.Western blot was used to detect RhoA,phosphorylated extracellular signal regulated kinase 1/2(p-ERK1/2),phosphorylated stress activated protein kinase 1(p-JNK1),and p-P38 protein in MOVAS cells.Results Compared with the control group,the OD450 value,scratch healing rate,OPN protein positive expression,PCNA,MMP-9,MMP-2 mRNA expression,RhoA,p-ERK1/2,p-JNK1,p-P38 protein expression of MOVAS cells in the Ang II group increased,while the positive expression of α-SMA protein decreased significantly(P<0.05).Compared with the Ang Ⅱ group,the OD450 value,scratch healing rate,OPN protein positive expression,PCNA,MMP-9,MMP-2 mRNA expression,and RhoA,p-ERK1/2,p-JNK1,and p-P38 proteins of MOVAS cells in the low,medium,and high dose groups of ginsenoside Re decreased,while the positive expression of α-SMA protein increased.The trend was most significant in the high dose group of ginsenoside Re(P<0.05).Compared with ginsenoside Re high-dose group,OD450 value,scratch healing rate,OPN protein positive expression,PCNA,MMP-9,MMP-2 mRNA expression and RhoA,p-ERK1/2,p-JNK1,p-P38 protein in ginsenoside Re high-dose+U46619 group increased,the positive expression of α-SMA protein decreased significantly(P<0.05).Conclusion Ginsenoside Re may inhibit the proliferation,migration,and phenotype transformation of Ang Ⅱ in MOVAS cells by suppressing the RhoA/MAPK pathway.

Objective To observe the value of ultrasound convolutional neural network(CNN)model for assisting distinguishment of cervical lymph node lymphoma and atypical reactive hyperplasia.Methods Totally 335 cases of cervical lymph node lymphoma and 335 cases of atypical reactive hyperplasia were retrospectively enrolled,including 520 cases in development group(260 cases of lymphoma and 260 cases of atypical reactive hyperplasia)and 150 cases in validation group(75 cases of lymphoma and 75 cases of atypical reactive hyperplasia).Patients in development group were divided into training set(182 cases of lymphoma and 182 cases of atypical reactive hyperplasia)and test set(78 cases of lymphoma and 78 cases of atypical reactive hyperplasia)at the ratio of 7∶3.One target lymph node was selected for each case,and one gray-scale ultrasound image and one CDFI were included for training,testing CNN models and verifying the auxiliary efficacy of the models.Based on ultrasound images in training set,5 CNN models,including AlexNet,VGG16,ResNet18,DenseNet161 and EfficientNet-B0,were constructed and trained for distinguishing cervical lymph node lymphoma and atypical reactive hyperplasia,and the models were tested in test set to screen out the best one with the highest classification accuracy.The efficacy of the best CNN model for assisting distinguishment of cervical lymph node lymphoma and atypical reactive hyperplasia was observed.Results Among 5 CNN models,the accuracy of ResNet18 model in test set was the highest(78.21％),and ResNet18 model was regarded as the best CNN model,its sensitivity,specificity and accuracy for assisting distinguishment of cervical lymph node lymphoma and atypical reactive hyperplasia were all higher than those of independent diagnosis made by ultrasound physicians(all P＜0.01).Conclusion The constructed ResNet18 model could be used to effectively assist differentiating cervical lymph node lymphoma and atypical reactive hyperplasia.

Endometrial carcinoma (EC) is one of the most common gynecologic malignancies, with the majority of patients diagnosed at an early stage. Surgery is the primary treatment modality, and postoperative risk stratification plays a key role in guiding adjuvant therapy decisions. Lymphovascular space invasion (LVSI) is an important pathological feature that affects the prognosis of patients and serves as one of the key factors in risk stratification. This review summarizes the prognostic significance of LVSI in patients with early-stage EC and its impact on the selection of adjuvant therapies, aiming to provide new insights for exploring precise and individualized diagnosis and treatment.

Objective:To construct a recombinant bioluminescent bacteriophage (HT7) targeting Escherichia coli, and evaluate its ability to identify Escherichia coli. Methods:Initially, pCRISPR-sg (1-10) and PFN-1000 plasmid strains were constructed by genetic engineering, and the most efficient small guild RNA (sgRNA) were screened by bilayer plate. By the gene editing technique, which comprised homologous recombination and clustered regularly interspaced short palin dromic repeats (CRISPR)-Cas system, the Nanoluc luciferase gene was integrated into the downstream non-coding region of 10A gene of T7 phage, to constructe the bioluminescent phage HT7 successfully. The difference of biological characteristics between HT7 phage and T7 phage was evaluated by plaque assay and liquid amplification assay. In addition, 51 strains of Escherichia coli, 20 strains of Klebsiella pneumoniae, 14 strains of Staphylococcus aureus, 6 strains of Enterococcus faecium, 5 strains of Enterococcus faecalis, 3 strains of Acinetobacter baumannii and 1 strain of Pseudomonas aeruginosa were collected and isolated to evaluate the limit of detection and specificity of HT7 phage. Results:Among the 10 CRISPR-targeted cleavage systems constructed, sgRNA8 exhibited the highest cleavage efficiency, with a cleavage rate of 0.18. After three rounds of recombination screening using the pCas9/pCRISPR/PFN-1000 triple-plasmid system, PCR validation yielded recombinant phage bands at 2 798 bp, indicating the successful construction of the HT7 phage. The recombinant phage showed significant differences in biological characteristics in terms of lysis efficiency ( P<0.001), one-step growth curve ( P=0.001), and infection multiplicity ( P=0.031). Both lysis burst time and log growth node were extended by 10 min, with the optimal infection multiplicity being 0.1. Clinical sample testing identified lysis of 6 strains of Escherichia coli within 4.5 h, while other strains remained unaffected, with detection of pathogenic bacteria below 10 CFU/ml. Conclusions:The developed pCas9/pCRISPR/PFN-1000 triple-plasmid editing system efficiently edits the bacteriophage genome. The constructed HT7 fluorescent bacteriophage enables the detection of Escherichia coli below 10 CFU/ml within 4.5 hours, demonstrating low detection limits and high detection specificity.

AIM:To investigate the effects of overexpressing α-Klotho(KL)in RAW264.7 cells stimulated by oxidative stress on the proliferation, migration, tube-formation and tight junction of human umbilical vein endothelial cells(HUVECs).METHODS:RAW264.7 cells were categorized into control, 4-hydroxynonenal(4HNE), and 4HNE+KL groups, with F4/80 expression assessed via immunofluorescence staining. Three groups of conditional media were prepared for HUVECs and culture divided into Mø-NC, Mø-4HNE, and Mø-4HNE+KL groups. Cell proliferation was evaluated using CCK8 assay, while scratch test and Transwell assays were employed to measure cell migration. Additionally, tube-formation assay was conducted to assess cell tubule formation, and Western blot assay was utilized to detect the protein expression levels of Claudin 5, Occludin and ZO 1.RESULTS:The results of immunofluorescence staining showed that the fluorescence intensity of F4/80 of RAW264.7 cells in the 4HNE group was significantly enhanced compared with the control group, while that of F4/80 in the 4HNE+KL group was significantly decreased compared with the 4HNE group(all P<0.05). The CCK8 assay results revealed a significant increase in the proliferation of HUVECs in the Mø-4HNE group compared with the Mø-NC group. Conversely, the proliferation of the Mø-4HNE+KL group exhibited a significant decrease compared with that in the Mø-4HNE group(all P<0.01). The results of scratch test and Transwell assays demonstrated a significant increase in the migration of HUVECs in the Mø-4HNE group compared with the Mø-NC group, while the migration of the Mø-4HNE+KL group exhibited a significant decrease compared with the Mø-4HNE group(all P<0.01). In the tube-formation assay, it was observed that the number of tubes formed by HUVECs in the Mø-4HNE group was significantly increased compared with the Mø-NC group, while that of tubes formed in the Mø-4HNE+KL group was significantly decreased compared with the Mø-4HNE group(all P<0.01). Additionally, the Western blot results revealed a significant decrease in the relative expression levels of Claudin 5, Occludin, and ZO 1 in the Mø-4HNE group compared with the Mø-NC group. Conversely, in the Mø-4HNE+KL group, there was a significant increase in the relative expression levels of Claudin 5, Occludin, and ZO 1 compared to the Mø-4HNE group(all P<0.01).CONCLUSIONS: KL inhibits the proliferation, migration, and tube-formation of HUVECs while enhancing the tight junction by changing the activation state of macrophages in the diabetic oxidative stress environment.

Objective To investigate the causal associations between components of metabolic syndrome(MetS)and endometrial carcinoma using Mendelian randomization(MR).Methods Data mining of the Genome-Wide Association Studies(GWAS)database was performed,with the exposure factors being MetS components(lipids,blood pressure,blood glucose,and obesity)and the outcome factor being endometrial carcinoma.MR analyses were performed with the help of regression models,including MR-Egger method,weighted median method,and inverse variance weighted(IVW)method,and the causal relationship between MetS components and endometrial carcinoma was evaluated by odds ratio(OR).Reverse MR analysis was performed for the MetS components found to have a causal relationship with endometrial carcinoma in the forward MR analysis.Results After applying Benjamini-Hochberg correction,IVW results showed a causal relationship between multiple MetS components(obesity,lipids,blood pressure,and blood glucose)and endometrial carcinoma.Specifically,the three components of obesity,including body mass index,overweight,and percentage of body fat,were causally associated with an increased risk of endometrial carcinoma(P＜0.001,OR＞1).In blood lipids,high cholesterol levels(P＜0.001,OR＜1),high triglyceride levels,and high phospholipid levels were causally associated with a reduced risk of endometrial carcinoma(P＜0.05,OR＜1).Regarding blood pressure,heart disease,atherosclerosis,and stroke were causally associated with a reduced risk of endometrial carcinoma(P＜0.05,OR＜1).Regarding blood glucose,low fasting insulin levels,type 1 diabetes mellitus,insulin resistance,and high glycated hemoglobin levels were causally associated with a reduced risk of endometrial carcinoma(P＜0.05,OR＜1),while type 2 diabetes mellitus and high fasting insulin levels were causally associated with an increased risk of endometrial carcinoma(P＜0.05,OR＞1).Reverse MR analysis did not produce any evidence for a reverse causality between the above positive MetS components and endometrial carcinoma.Conclusion The MR study suggests that obesity and type 2 diabetes mellitus are risk factors for endometrial carcinoma,while other MetS components,including hyperlipidemia,cardiovascular diseases,insulin resistance,and complications of diabetes mellitus,are protective factors for endometrial carcinoma.Further research is needed to clarify the association between MetS and endometrial carcinoma and to further explore the underlying mechanisms involved.

Objective: To investigate the correlation between tumor size, tumor location, and prognosis in patients with early-stage endometrial cancer (EC) receiving adjuvant radiotherapy. Methods: Data of patients who had been treated for stage I–II EC from March 1999 to September 2017 in 13 tertiary hospitals in China was screened. Cox regression analysis was performed to investigate associations between tumor size, tumor location, and other clinical or pathological factors with cancer-specific survival (CSS) and distant metastasis failurefree survival (DMFS). The relationship between tumor size as a continuous variable and prognosis was demonstrated by restricted cubic splines. Prognostic models were constructed as nomograms and evaluated by Harrell’s C-index, calibration curves and receiver operating characteristic (ROC) curves. Results: The study cohort comprised 805 patients with a median follow-up of 61 months and a median tumor size of 3.0 cm (range 0.2–15.0 cm). Lower uterine segment involvement (LUSI) was found in 243 patients (30.2%). Tumor size and LUSI were identified to be independent prognostic factors for CSS. Further, tumor size was an independent predictor of DMFS. A broadly positive relationship between poor survival and tumor size as a continuous variable was visualized in terms of hazard ratios. Nomograms constructed and evaluated for CSS and DMFS had satisfactory calibration curves and C-indexes of 0.847 and 0.716, respectively. The area under the ROC curves for 3- and 5-year ROC ranged from 0.718 to 0.890. Conclusion Tumor size and LUSI are independent prognostic factors in early-stage EC patients who have received radiotherapy. Integrating these variables into prognostic models would improve predictive ability.