AIM: To develop a novel gene-delivery therapeutic based on CRISPR/Cas9 genome editing technology capable of specifically targeting and knocking out the VEGFA gene, thereby achieving sustained suppression of VEGFA expression in retinal pigment epithelial(RPE)cells and providing a new strategy for gene therapy in retinal neovascular diseases.METHODS:Single guide RNAs targeting the human VEGFA gene for knockout were designed, and corresponding recombinant plasmids were constructed. A novel polymer(PTEE)was used to encapsulate the plasmids to prepare a PTEE-loaded anti-VEGFA plasmid(PLAP)gene delivery system. PTEE materials at concentrations of 0.1, 0.2, 0.4, 0.8, and 1.6 μg/μL were co-incubated with ARPE-19 cells, and the biocompatibility of PTEE was evaluated using the cell counting kit-8(CCK-8)assay. Recombinant plasmids expressing green fluorescent protein(GFP)were constructed. Lipofectamine 3000 and jetOPTIMUS®DNA transfection reagents were used as control groups, and PTEE nanomaterials were used as the experimental group to encapsulate the plasmids. When the cell confluence reached 80%, the formulations were transfected into ARPE-19 and 293T cells. GFP expression was observed under light microscopy, and the transfection efficiencies of each group were compared. ARPE-19 cells were induced under hypoxia, and PLAP was transfected into the cells. The expression level of VEGFA was detected by enzyme-linked immunosorbent assay(ELISA)to evaluate the efficacy of this novel gene delivery system.RESULTS: After co-incubation of ARPE-19 cells with different concentrations of PTEE for 24 h and 48 h, no significant effect on cell viability was observed in any group. The transfection efficiency of PLAP in ARPE-19 cells was higher than that in the Lipo3000 and jetOPTIMUS groups, with statistically significant differences(P<0.01). Hypoxia for 6 h significantly induced the upregulation of VEGFA mRNA expression in ARPE-19 cells, and under hypoxic conditions, the PTEE group exhibited a significant inhibitory effect on VEGFA expression(P<0.01).CONCLUSION:PLAP exhibits favorable biocompatibility and prominent VEGFA inhibitory effects in vitro, making it a potential candidate drug for gene therapy of retinal neovascular diseases.

Objective To explore the application values of different detection methods in monitoring the decline pattern of syphilis-spe-cific antibody in the non-congenital syphilis children.Methods A total of 80 non-congenital syphilis children were included in the study.The serum specimens were collected after birth,and the syphilis-specific antibodies were detected using electrochemilumines-cence immunoassay(ECLIA),western blotting(WB),treponema pallidum particle agglutination assay(TPPA),enzyme-linked im-munosorbent assay(ELISA),and toluidine red unheated serum test(TRUST).Follow-up was conducted every three months until the positive results of ELISA and TRUST turned to negative.Results The results of ECLIA showed that the syphilis-specific antibody lev-els in the non-congenital syphilis children declined to 25％of the level at birth within 2 to 3 months,and the rate of decline was inde-pendent of the initial concentration.WB analysis indicated that the specific IgG bands in non-congenital syphilis children at birth were consistent with those of their mother,and the sequence of specific antibodies decline was as follows:TPN47,TPN15,TPN45,and TPN17.Due to methodological limitations,the absorbance values of ELISA showed no significant change during the first three months after birth when high concentrations of antibodies were present in the samples,but it showed high sensitivity in the detection for the samples with low-concentration of syphilis antibodies.The detection rates of ECLIA,TPPA,and WB were compared by using ELISA as the reference method.At birth,the detection rates of syphilis antibodies were 100％,100％,and 90％,respectively.In 3 months after birth,the detection rates were 100％,100％,and 75％.In 6 months after birth,,they were 100％,46％,and 15％.In 9 months after birth,they were 83％,33％,and 0％.The positive rate of TRUST was 17.5％at birth.and turned to negative in 3 month of follow-up.Conclusion Syphilis specific IgG antibodies may fully transferred to the fetus and decline in a predictable pattern after birth.The comprehensive analysis for the results of the four methods suggested that dynamic detection using ECLIA method could be used to pre-dict the risk of non-congenital syphilis or terminate the follow-up at 3 months,while the seroconversion detected by WB was earlier than that by TPPA,while ELISA required the longest follow-up period.

Objective To establish and validate a machine learning model integrating abdominal contrast-enhanced CT radiomics features and clinical characteristics,and to construct a predictive model for the efficacy of anticoagulant treatment in portal vein thrombosis(PVT).Methods A retrospective selection was conducted on 94 PVT patients who received anticoagulant treatment.Patients were divided into effective and ineffective treatment groups based on the follow-up results.Clinical information was collected,and imaging features were evaluated.Univariate and multivariate logistic regression were performed to select clinical information and imaging fea-tures for constructing a clinical-imaging model.On CT venous phase images,the PVT mask was delineated and radiomics features were extracted,and the radiomics model was screened and established.A combined model was further developed using features from both the clinical-imaging and radiomics models.Receiver operating characteristic(ROC)curves were used to evaluate the predictive efficacy of different models.Results The area under the curve(AUC)for the clinical-imaging model,radiomics model,and com-bined model were 0.594,0.794,and 0.776,respectively.The radiomics and combined models demonstrated superior predictive efficacy for anticoagulant treatment in PVT compared to the clinical-imaging model.No significant difference in performance was observed between radiomics and combined models.Conclusion The radiomics model and combined model based on abdominal contrast-enhanced CT can effectively predict the efficacy of anticoagulant treatment for PVT.

Objective To explore the safety and feasibility of extracorporeal suture traction of round ligament of the uterus assisted two-port laparoscopic internal ring closure in the treatment of pediatric ovarian sliding hernia.Methods From September 2019 to September 2024,57 children with ovarian sliding hernia underwent extracorporeal suture traction of round ligament of the uterus assisted two-port laparoscopic internal ring closure.Under the guidance of single-port umbilical laparoscopic monitoring,a 2-0 suturing needle with thread was inserted through the opposite internal ring and used to suture the round ligament of the uterus at the affected side's internal ring.The needle was passed through and exited near the entry point,and the extracorporeal suture traction was used to pull the ovary(and part of the fallopian tube)back into the abdominal cavity from the inguinal canal,maintaining the suture tension to prevent the ovary and fallopian tube from re-entering the internal ring.The internal ring was fully exposed.A 2-0 suturing needle with thread was inserted at the affected side(2-3 cm from the internal ring)to perform a circular suture of the peritoneum at the internal ring.The internal ring was closed at the level of internal ring orifice.Results All the 57 cases of ovarian sliding hernia were confirmed to be unilateral,with 18 cases on the right side(including 12 cases of pure ovarian sliding hernia and 6 cases of ovarian and fallopian tube sliding hernia)and 39 cases on the left side(including 22 cases of pure ovarian sliding hernia and 17 cases of ovarian and fallopian tube sliding hernia).The average surgery time was(22.5±2.5)min.All the patients were discharged within 6 h postoperatively and no complications were noted.A total of 49 patients were followed up for6-20 months(mean 9.5 months),with 21 cases followed for more than 12 months.No complications such as incision infection,recurrence of hernia,or Nuck's cyst were observed.Ultrasonic examinations showed normal bilateral ovarian blood flow.Conclusion For pediatric ovarian sliding hernia,extracorporeal suture traction of round ligament of the uterus assisted two-port laparoscopic internal ring closure can effectively retract the ovary(and part of the fallopian tube)from the inguinal canal back into the abdominal cavity,fully exposing the normal anatomical form of the internal ring,with the characteristics of safety and feasibility.

Objective To explore the application values of different detection methods in monitoring the decline pattern of syphilis-spe-cific antibody in the non-congenital syphilis children.Methods A total of 80 non-congenital syphilis children were included in the study.The serum specimens were collected after birth,and the syphilis-specific antibodies were detected using electrochemilumines-cence immunoassay(ECLIA),western blotting(WB),treponema pallidum particle agglutination assay(TPPA),enzyme-linked im-munosorbent assay(ELISA),and toluidine red unheated serum test(TRUST).Follow-up was conducted every three months until the positive results of ELISA and TRUST turned to negative.Results The results of ECLIA showed that the syphilis-specific antibody lev-els in the non-congenital syphilis children declined to 25％of the level at birth within 2 to 3 months,and the rate of decline was inde-pendent of the initial concentration.WB analysis indicated that the specific IgG bands in non-congenital syphilis children at birth were consistent with those of their mother,and the sequence of specific antibodies decline was as follows:TPN47,TPN15,TPN45,and TPN17.Due to methodological limitations,the absorbance values of ELISA showed no significant change during the first three months after birth when high concentrations of antibodies were present in the samples,but it showed high sensitivity in the detection for the samples with low-concentration of syphilis antibodies.The detection rates of ECLIA,TPPA,and WB were compared by using ELISA as the reference method.At birth,the detection rates of syphilis antibodies were 100％,100％,and 90％,respectively.In 3 months after birth,the detection rates were 100％,100％,and 75％.In 6 months after birth,,they were 100％,46％,and 15％.In 9 months after birth,they were 83％,33％,and 0％.The positive rate of TRUST was 17.5％at birth.and turned to negative in 3 month of follow-up.Conclusion Syphilis specific IgG antibodies may fully transferred to the fetus and decline in a predictable pattern after birth.The comprehensive analysis for the results of the four methods suggested that dynamic detection using ECLIA method could be used to pre-dict the risk of non-congenital syphilis or terminate the follow-up at 3 months,while the seroconversion detected by WB was earlier than that by TPPA,while ELISA required the longest follow-up period.

Objective To explore the safety and feasibility of 3-mm laparoscopic needle holder combined with 3-0 needle-thread nylon in external silk thread traction-assisted two-port laparoscopic internal ring suturing for the treatment of inguinal indirect hernia in children≤3 months old.Methods From January 2020 to September 2024,49 children ≤ 3 months old with indirect hernia underwent external silk thread traction-assisted two-port laparoscopic internal ring suturing.All the patients were found to complicated with redundant peritoneum around internal ring.Under transumbilical single-port laparoscopic monitoring,the 3-mm laparoscopic needle holder holing the 3-0 needle-thread nylon was inserted through the inferior edge of the internal ring,entering the peritoneum and traveling through the retroperitoneal space.The external nylon thread was advanced to drag the redundant peritoneum,allowing the suture needle cross the surface of the spermatic cord vessels,the Doom's triangle,and the vas deferens(boys)or the uterine round ligament(girls)in the retroperitoneal space,followed by circular suturing and closure of the internal ring.Results The surgery time was(13.5±2.5)min for the 35 cases of unilateral hernia and(24.5±3.5)min for the 14 cases of bilateral hernia.No intraoperative complications occurred,and all the patients were discharged within 6 h post-operation.Except for 3 children of lost to follow-up,46 children were followed up for 6-26 months(mean,9.5 months),with 19 cases followed for＞12 months.No complications such as recurrence,wound infection,hydrocele,iatrogenic undescended testicle,or testicular atrophy were observed during the follow-ups.Conclusion For children ≤ 3 months old with indirect hernia,the use of 3-mm laparoscopic needle holder combined with 3-0 needle-thread nylon in external silk thread traction-assisted two-port laparoscopic internal ring suturing enhances the flexibility and convenience of the procedure,being safety and feasibility.

Objective To explore the safety and feasibility of extracorporeal suture traction of round ligament of the uterus assisted two-port laparoscopic internal ring closure in the treatment of pediatric ovarian sliding hernia.Methods From September 2019 to September 2024,57 children with ovarian sliding hernia underwent extracorporeal suture traction of round ligament of the uterus assisted two-port laparoscopic internal ring closure.Under the guidance of single-port umbilical laparoscopic monitoring,a 2-0 suturing needle with thread was inserted through the opposite internal ring and used to suture the round ligament of the uterus at the affected side's internal ring.The needle was passed through and exited near the entry point,and the extracorporeal suture traction was used to pull the ovary(and part of the fallopian tube)back into the abdominal cavity from the inguinal canal,maintaining the suture tension to prevent the ovary and fallopian tube from re-entering the internal ring.The internal ring was fully exposed.A 2-0 suturing needle with thread was inserted at the affected side(2-3 cm from the internal ring)to perform a circular suture of the peritoneum at the internal ring.The internal ring was closed at the level of internal ring orifice.Results All the 57 cases of ovarian sliding hernia were confirmed to be unilateral,with 18 cases on the right side(including 12 cases of pure ovarian sliding hernia and 6 cases of ovarian and fallopian tube sliding hernia)and 39 cases on the left side(including 22 cases of pure ovarian sliding hernia and 17 cases of ovarian and fallopian tube sliding hernia).The average surgery time was(22.5±2.5)min.All the patients were discharged within 6 h postoperatively and no complications were noted.A total of 49 patients were followed up for6-20 months(mean 9.5 months),with 21 cases followed for more than 12 months.No complications such as incision infection,recurrence of hernia,or Nuck's cyst were observed.Ultrasonic examinations showed normal bilateral ovarian blood flow.Conclusion For pediatric ovarian sliding hernia,extracorporeal suture traction of round ligament of the uterus assisted two-port laparoscopic internal ring closure can effectively retract the ovary(and part of the fallopian tube)from the inguinal canal back into the abdominal cavity,fully exposing the normal anatomical form of the internal ring,with the characteristics of safety and feasibility.

Objective To establish and validate a machine learning model integrating abdominal contrast-enhanced CT radiomics features and clinical characteristics,and to construct a predictive model for the efficacy of anticoagulant treatment in portal vein thrombosis(PVT).Methods A retrospective selection was conducted on 94 PVT patients who received anticoagulant treatment.Patients were divided into effective and ineffective treatment groups based on the follow-up results.Clinical information was collected,and imaging features were evaluated.Univariate and multivariate logistic regression were performed to select clinical information and imaging fea-tures for constructing a clinical-imaging model.On CT venous phase images,the PVT mask was delineated and radiomics features were extracted,and the radiomics model was screened and established.A combined model was further developed using features from both the clinical-imaging and radiomics models.Receiver operating characteristic(ROC)curves were used to evaluate the predictive efficacy of different models.Results The area under the curve(AUC)for the clinical-imaging model,radiomics model,and com-bined model were 0.594,0.794,and 0.776,respectively.The radiomics and combined models demonstrated superior predictive efficacy for anticoagulant treatment in PVT compared to the clinical-imaging model.No significant difference in performance was observed between radiomics and combined models.Conclusion The radiomics model and combined model based on abdominal contrast-enhanced CT can effectively predict the efficacy of anticoagulant treatment for PVT.

Objective To explore the safety and feasibility of 3-mm laparoscopic needle holder combined with 3-0 needle-thread nylon in external silk thread traction-assisted two-port laparoscopic internal ring suturing for the treatment of inguinal indirect hernia in children≤3 months old.Methods From January 2020 to September 2024,49 children ≤ 3 months old with indirect hernia underwent external silk thread traction-assisted two-port laparoscopic internal ring suturing.All the patients were found to complicated with redundant peritoneum around internal ring.Under transumbilical single-port laparoscopic monitoring,the 3-mm laparoscopic needle holder holing the 3-0 needle-thread nylon was inserted through the inferior edge of the internal ring,entering the peritoneum and traveling through the retroperitoneal space.The external nylon thread was advanced to drag the redundant peritoneum,allowing the suture needle cross the surface of the spermatic cord vessels,the Doom's triangle,and the vas deferens(boys)or the uterine round ligament(girls)in the retroperitoneal space,followed by circular suturing and closure of the internal ring.Results The surgery time was(13.5±2.5)min for the 35 cases of unilateral hernia and(24.5±3.5)min for the 14 cases of bilateral hernia.No intraoperative complications occurred,and all the patients were discharged within 6 h post-operation.Except for 3 children of lost to follow-up,46 children were followed up for 6-26 months(mean,9.5 months),with 19 cases followed for＞12 months.No complications such as recurrence,wound infection,hydrocele,iatrogenic undescended testicle,or testicular atrophy were observed during the follow-ups.Conclusion For children ≤ 3 months old with indirect hernia,the use of 3-mm laparoscopic needle holder combined with 3-0 needle-thread nylon in external silk thread traction-assisted two-port laparoscopic internal ring suturing enhances the flexibility and convenience of the procedure,being safety and feasibility.

Objective To analyze the seroepidemiology characteristics of Epstein-Barr virus(EBV)infection in children in Shenyang.Methods From June 2022 to May 2023,serum was collected from 12 083 children at Shengjing hospital of China Medical University.Serum capsid antigen(VCA)-IgM,VCA-IgG,EBV nuclear antigen-IgG(EBNA-IgG),and early antigen-IgG(EA-IgG)antibodies were detected using the LIAISION chemiluminescence immunoassay.EBV-DNA was detected using real-time PCR.Differences in antibody positivity rates between sexes,ages,and seasons of onset were compared.Results In 12 083 patients,the positive rates of VCA-IgM,VCA-IgG,EBNA-IgG,and EA-IgG were 9.95％(1 202/12 083),50.57％(6 110/12 083),46.03％(5 562/12 083)and 4.93％(596/12 083).The positive rates of VCA-IgM and EA-IgG were lower in male patients(9.09％and 4.44％,respectively)than in female patients(11.10％,5.60％;all P＜0.05).The differences in the positive rates for VCA-IgM,VCA-IgG,EBNA-IgG,and EA-IgG antibodies in children of different ages were statistically significant(all P＜0.05).Differences in the positive rates of VCA-IgG,EBNA-IgG,and EA-IgG antibodies were statistically significant when compared between seasons(all P＜0.05).Fourteen EBV antibody-positive combinations were detected,of which the main combination was VCA-IgG and EBNA-IgG double-positive,with a total of 4 741 cases(39.24％).Of the 3 712 children who underwent EBV-DNA detection testing,3 034(81.73％)were EBV-DNA-negative and 678(18.27％)were EBV-DNA-posi-tive.VCA-IgG and EBNA-IgG double positivity was the most common in EBV-DNA-negative and EBV-DNA-positive children,accounting for 983(26.48％)and 194 cases(5.23％),respectively.Conclusion Both VCA-IgG and EBNA-IgG antibodies are main positive in children with EBV infection in Shenyang.The positive rate of EBV antibodies is lower in boys than in girls.The positive rates of EBV anti-bodies differ in children of different ages and seasons of onset.