Objective To analyze the influencing factors of tumor-specific mortality in patients with advanced endometrial carcinoma and construct a risk prediction model.Methods A total of 150 patients with advanced endometrial carcinoma admitted to the Fourth Clinical College of Zhejiang Chinese Medical University,Hangzhou First People's Hospital from January 2020 to January 2023 were selected as subjects.Cox regression was employed to analyze factors influencing mortality in advanced endometrial carcinoma.A Logistic regression model was constructed,with receiver operating characteristic(ROC)curve used to evaluate model discrimination,calibration curve for precision assessment,and decision curve for net benefit evaluation.Results Overall 2-year survival rate of 150 patients was 58.67％.The univariate analysis demonstrated that histological grade,lymph node metastasis,tumor maximum diameter,and preoperative carcinoembryonic antigen 125(CA125)levels were all significant risk factors for mortality in advanced endometrial carcinoma(P＜0.05).Multivariate analysis confirmed these parameters as independent prognostic indicators(P＜0.05).A Logistic regression model established using multivariate Cox regression yielded a C index of 0.873(95％CI:0.792-0.951).ROC curve analysis revealed the model's area under the curve for predicting mortality risk factors in advanced endometrial carcinoma was 0.916(95％CI:0.872-0.960).Conclusion The linear chart prediction model constructed based on histological grading,lymph node metastasis,maximum tumor diameter and preoperative CA125 level in this study can effectively predict the death of advanced endometrial carcinoma.

Objective To analyze the influencing factors of tumor-specific mortality in patients with advanced endometrial carcinoma and construct a risk prediction model.Methods A total of 150 patients with advanced endometrial carcinoma admitted to the Fourth Clinical College of Zhejiang Chinese Medical University,Hangzhou First People's Hospital from January 2020 to January 2023 were selected as subjects.Cox regression was employed to analyze factors influencing mortality in advanced endometrial carcinoma.A Logistic regression model was constructed,with receiver operating characteristic(ROC)curve used to evaluate model discrimination,calibration curve for precision assessment,and decision curve for net benefit evaluation.Results Overall 2-year survival rate of 150 patients was 58.67％.The univariate analysis demonstrated that histological grade,lymph node metastasis,tumor maximum diameter,and preoperative carcinoembryonic antigen 125(CA125)levels were all significant risk factors for mortality in advanced endometrial carcinoma(P＜0.05).Multivariate analysis confirmed these parameters as independent prognostic indicators(P＜0.05).A Logistic regression model established using multivariate Cox regression yielded a C index of 0.873(95％CI:0.792-0.951).ROC curve analysis revealed the model's area under the curve for predicting mortality risk factors in advanced endometrial carcinoma was 0.916(95％CI:0.872-0.960).Conclusion The linear chart prediction model constructed based on histological grading,lymph node metastasis,maximum tumor diameter and preoperative CA125 level in this study can effectively predict the death of advanced endometrial carcinoma.

Pseudorabies virus(PRV)is the etiological agent of pseudorabies in pigs,which is char-acterized by dyspnea,reproductive disorders,and neurological diseases,and it spreads widely a-round the world.Since 2011,the newly emerged PRV variants have resulted in poor immunity pro-tection of traditional vaccine strains,and the original method of vaccine strain preparation is time-consuming and labor-intensive.Therefore,it is urgently needed to develop an efficient screening method of the vaccine strain at present.Using CRISPR/Cas9 gene editing technology in this study,two single guide RNAs(sgRNA)were designed targeting the virulence gene TK of PRV variant strain CH/JX/2016,and then the enhanced green fluorescent protein the reporter(EGFP)gene was inserted at the TK locus by a homologous repair plasmid.After multiple rounds of plaque puri-fication,the rPRV-ΔTK/EGFP strain was obtained.The results showed the cleavage efficiency of the two sgRNAs was extremely high.The preparation of rPRV-ΔTK/EGFP strain was succeed af-ter only three rounds of purification,and the EGFP expressed normally.The CRISPR/Cas9 system can edit the PRV gene simply,rapidly,and efficiently,and exhibits great potential in the construction of vaccine candidate strains.Meanwhile,the rescued rPRV-ΔTK/EGFP strain not only could be used as a tracer strain in PRV variant infection progresses,but also for subsequent antivi-ral drug screening.

Genicular nerve block is usually used for the treatment of chronic pain of knee osteoar-thritis,which can effectively relieve knee pain and preservemotor function.With the rapid development of ultrasound technology,ultrasound-guided genicular nerve block can improve the accuracy of nerve block and reduce block-related complications.This article reviews the research progress of ultrasound-guided genicular nerve block in three aspects:anatomy,operation methods and clinical application.

Objective To establish a coronavirus(CoV)infection model using human nasal mucosa organoids for testing antiviral drugs and evaluate the feasibility of using human nasal mucosa organoids with viral infection as platforms for viral research and antiviral drug development.Methods Human nasal mucosa organoids were tested for susceptibility to SARS-CoV-2 and HCoV-OC43 pseudoviruses.In a P3 laboratory,nasal mucosa organoids were infected with the original strain of SARS-CoV-2 and 4 variant strains,and the infection conditions were optimized.The viral loads in the culture supernatants were measured at different time points using RT-qPCR,and immunofluorescence assay was employed to localize SARS-CoV-2 nucleocapsid protein to determine the type of the infected cells.In the optimized nasal mucosa viral infection model,the antiviral effects of camostat and bergamot extract(which were known to inhibit SARS-CoV-2)were tested and the underlying molecular mechanisms were explored.Results In the optimized nasal mucosa organoid models infected with SARS-CoV-2 and HCoV-OC43 pseudoviruses,the viral load in the culture supernatants increased significantly during the period of 2 to 24 h following the infection,which confirmed infection of the organoids by both of the pseudoviruses.The nasal mucosa organoids could be stably infected by the original SARS-CoV-2 strain and its 4 variant strains,validating successful establishment of the viral infection model,in which both camostat and bergamot extract exhibited dose-dependent antiviral effects.Conclusions Human nasal mucosa organoids with SARS-CoV-2 infection can serve as platforms for screening and testing antiviral drugs,particularly those intended for nasal administration.

Objective To compare the effectiveness of different β-blockers for preventing postoperative atrial fibrillation (POAF) after cardiac surgery. Methods Databases of PubMed, Science Direct, Web of Science, The Cochrane Library, SinoMed, CKNI, VIP, WanFang were searched by the computer from inception to April 31, 2022 to collect randomized controlled studies on the comparison of the effectiveness of different β-blockers for preventing POAF. Two investigators independently screened the literature and extracted information. The quality of the literature was evaluated using Cochrane bias risk tool, and RevMan 5.3 and STATA 17.0 were used for meta-analysis. Results A total of 17 randomized controlled studies with 3 290 patients were included. Direct meta-analysis showed that landiolol and metoprolol were more effective than placebo (P≤0.001), and carvedilol was more effective than metoprolol in preventing the development of POAF (P<0.001). Network meta-analysis showed that landiolol, carvedilol and metoprolol were more effective than placebo in preventing the incidence of POAF (P<0.05). Landiolol and carvedilol were more effective than metoprolol, and carvedilol were more effective than nebivolol (P<0.05). The surface under the cumulative ranking curve from high to low were carvedilol, landilol, propranolol, atenolol, metoprolol, and nebivolol. Conclusion Carvedilol and landilol have different degrees of improvement in the occurrence of POAF, and carvedilol has the best preventive effect. More studies are required to verify the strength of evidence due to the limited sample size.

Objective To explore the effects of image registration using various regions of interest(ROI)on the setup error for nasopharyngeal carcinoma(NPC)patients who were immobilized individually.Methods Forty-three NPC patients who required radiotherapy were enrolled.The patients were immobilized with customized plastic foam and thermoplastic mask,and CBCT verification was performed once a week.In CBCT images,ROI was divided into the whole ROI(ROIPTV)and 7 local ROI containing different cervical structures(ROIsphenoid sinus,ROIatlantoaxial,ROIneck3,ROIneck4,ROIneck5,ROIneck6,and ROIneck7),which were then used for registrations with localized CT image.The setup errors in superior-inferior(SI),left-right(LR),anterior-posterior(AP),Pitch,Roll,and Yaw directions were recorded.Results In SI direction,the setup errors within 0.3 cm accounted for 89.74%for ROIneck7,and more than 90%for the other ROI.The proportion of setup errors within 0.3 cm gradually increased with the neck upward in LR direction,and they were 76.78%,81.70%,85.26%,and above 90%for ROIneck7,ROIneck6,ROIneck5,and the other ROI,respectively.In AP direction,the proportions of setup errors within 0.3 cm were less than 90%,except for ROIatlantoaxial and ROIneck3.The setup errors of ROIsphenoid sinus,ROIatlantoaxial,ROIneck3,and ROIneck4 were significantly positively correlated with ROIPTV in SI direction,and the correlation coefficients(R)were 0.94,0.95,0.90,and 0.83,respectively.In LR direction,there were positive correlations between the setup errors of ROIatlantoaxial and ROIsphenoid sinus(R=0.95),ROIneck3 and ROIsphenoid sinus(R=0.91),ROIPTV and ROIneck3(R=0.91).The setup errors of ROIPTV in AP direction were positively correlated with ROIatlantoaxial vertebrae and ROIneck3(R=0.88,0.90).The margins of all ROIs ranged from 0.38 cm to 1.01 cm.The extension of ROIneck6 and ROIneck7 in AP direction exceeded 0.9 cm,and the extension of ROIneck7 reached 0.95 cm in SI direction.Conclusion ROIPTV and ROIsphenoid sinus,ROIatlantoaxial,ROIneck3 are significantly correlated in SI,LR,and AP directions.The setup error of nasopharyngeal carcinoma patients gradually increases with the neck down.The nasopharyngeal and cervical regions need to be expanded in segments when patients are immobilized individually.

Objective To establish a coronavirus(CoV)infection model using human nasal mucosa organoids for testing antiviral drugs and evaluate the feasibility of using human nasal mucosa organoids with viral infection as platforms for viral research and antiviral drug development.Methods Human nasal mucosa organoids were tested for susceptibility to SARS-CoV-2 and HCoV-OC43 pseudoviruses.In a P3 laboratory,nasal mucosa organoids were infected with the original strain of SARS-CoV-2 and 4 variant strains,and the infection conditions were optimized.The viral loads in the culture supernatants were measured at different time points using RT-qPCR,and immunofluorescence assay was employed to localize SARS-CoV-2 nucleocapsid protein to determine the type of the infected cells.In the optimized nasal mucosa viral infection model,the antiviral effects of camostat and bergamot extract(which were known to inhibit SARS-CoV-2)were tested and the underlying molecular mechanisms were explored.Results In the optimized nasal mucosa organoid models infected with SARS-CoV-2 and HCoV-OC43 pseudoviruses,the viral load in the culture supernatants increased significantly during the period of 2 to 24 h following the infection,which confirmed infection of the organoids by both of the pseudoviruses.The nasal mucosa organoids could be stably infected by the original SARS-CoV-2 strain and its 4 variant strains,validating successful establishment of the viral infection model,in which both camostat and bergamot extract exhibited dose-dependent antiviral effects.Conclusions Human nasal mucosa organoids with SARS-CoV-2 infection can serve as platforms for screening and testing antiviral drugs,particularly those intended for nasal administration.

Objective To evaluate the effects of long voyage on crew's cardiac function.Methods A total of 47 crew members from a shipyard during the maintenance period of two ships with the same type from October 2017 to April 2018 were selected as research subjects.They were divided into experimental group(n=24)and control group(n=23).The first test was performed in all subjects within 5 days of enrollment.The experimental group participated in a 34-day seagoing voyage after the first test,while the control group continued to live on land and do regular physical exercise.All the subjects were tested again within 5 days after sailing.The submaximal exercise test was conducted according to the standard Bruce protocol.High definition impedance cardiogram was synchronously used to record heart rate and stroke output(SV)at rest.SV was continuously recorded to obtain its maximum value(SVmax)and the change of SV during exercise was analyzed.The exercise time,SV threshold,and SV threshold time were also recorded.Results There were no significant differences in the age,height,or weights at the beginning or end of the study between the two groups(P>0.05).At the end of the study,the exercise time of the experimental group was significantly shorter than that of the control group and the control group was prolonged(P<0.01).There was no significant difference in the SV threshold time,HR at rest,average resting SV,or SVmax between the two groups at enrollment(P>0.05).The SV threshold time at the end of the study was significantly shorter than that at enrollment in the experimental group,and there was a significant difference in the SV threshold time at the end of the study between the two groups(P<0.01).At the end of the study,the resting HR of the experimental group was significantly higher than that at enrollment and that of the control group(P<0.05);there were significant differences in the mean SV at rest and SVmax between groups and between intra-groups(P<0.05).Conclusion Long voyage can reduce the aerobic capacity and cardiac reserve of crew,and the preservation of aerobic exercise can improve the cardiac function and cardiac reserve.

Objective:To evaluate the status of T, B and NK lymphocytes in peripheral blood of patients with chronic hepatitis B virus(HBV) infection and low-level viremia after nucleos(t)ide analogue (NA) treatment and to provide ideas for solving low-level viremia.Methods:This retrospective study involved 344 patients with chronic HBV infection who had been treated with NAs. They were divided into two groups: low-level viremia group (LLV group) and complete virological response group (CVR group). Clinical data including basic information, biochemistry and coagulation test results, HBV DNA, peripheral blood lymphocyte counts, PD1 and CD28 expression by T lymphocytes, and perforin and granzyme B expression by NK lymphocytes were collected and compared between the two groups. Propensity matching analysis was performed to verify the accuracy of the results.Results:Among the 344 cases, 162 were in the LLV group and 182 in the CVR group. There were no significant differences in disease diagnosis, alanine aminotransferase (ALT), aspartate aminotransferase (AST) or albumin (ALB) level between the two groups ( P>0.05), but the differences in gender and age were statistically significant ( P<0.05). The differences in the counts and percentages of peripheral blood CD3 +, CD4 + and CD8 + T lymphocyte and CD4 + /CD8 + ratios between the two groups were not statistically significant ( P>0.05), but the expression of PD1 and CD28 by peripheral blood CD3 +, CD4 + and CD8 + T lymphocytes was higher in the LLV group than in the CVR group ( P<0.05). The count of peripheral blood CD19 + B lymphocytes in the LLV group was higher than that in the CVR group ( P>0.05), and the percentage of peripheral blood CD19 + B lymphocytes was also higher in the LLV group ( P<0.05). The count of peripheral blood CD16 + CD56 + NK lymphocytes and the expression of perforin in the LLV group were lower than those in the CVR group ( P>0.05). The percentage of peripheral blood CD16 + CD56 + NK lymphocytes and the expression of granzyme B in the LLV group were lower than those in the CVR group ( P<0.05). After propensity score matching, 108 cases in the LLV group and 108 cases in the CVR group showed no significant differences in basic information ( P>0.05); the percentage of CD4 + T lymphocytes and CD4 + /CD8 + ratio in peripheral blood T lymphocyte subsets were higher in the LLV group than in the CVR group, while the percentage of CD8 + lymphocytes was lower in the LLV group ( P<0.05); the expression of PD1 and CD28 by CD3 +, CD4 + and CD8 + T lymphocytes remained higher in the LLV group ( P<0.05); the differences in the counts and percentages of peripheral blood CD19 + B lymphocytes as well as CD16 + CD56 + NK lymphocytes between the two groups were not statistically significant ( P>0.05); no significant difference in the expression of perforin by CD16 + CD56 + NK lymphocytes was found between the two groups ( P>0.05), and the expression of granzyme B remained lower in the LLV group ( P<0.05). Conclusions:Abnormal number and function of T lymphocytes and decreased function of NK lymphocytes might be related to the development of LLV in patients with chronic HBV infection after treatment. Therefore, in addition to adjusting NAs, targeting of T and NK lymphocytes might also be a feasible measure for future LLV treatment.