Aim To construct a model of vascular endothelial cell(EC)-specific gene knockout mice of tumor necrosis factor receptor-associated factor 2(Traf2)by using Cre-loxP technolo-gy,thus to provide basis for the research of vascular dysfunction-related diseases related to the regulation of vascular EC activity through TRAF2.Methods The Cre-loxP system was used to construct a mouse model with EC-specific knockout of Traf2 gene(Traf2flox/flox Tek-iCre+).The mouse genotype was identi-fied through PCR and gel electrophoresis.Primary vascular ECs were isolated from the mice using flow cytometry.The knockout efficiency of Traf2 in vascular ECs was validated by Western blot and immunofluorescence.The pathological changes in blood ves-sels and major organs of the mice were examined using hematox-ylin-eosin(HE)staining.The tube-forming ability of primary vascular EC was assessed using Matrigel tube formation.Results The knockout mice met the required genetic criteria.Flow cy-tometry results demonstrated the successful isolation of primary vascular EC,and TRAF2 expression in vascular EC of knockout mice was significantly reduced(P＜0.01).Histological results showed that TRAF2 expression in the vessel of knockout mice decreased,and the morphology had no significant changes in their blood vessels and major organs.The Matrigel tube forma-tion demonstrated that the tube-forming ability of primary vascu-lar EC from knockout mice was reduced.Conclusion Traf2 specific knockout mouse model in vascular ECs is successfully constructed,providing a reliable animal model for research into the regulatory mechanisms of TRAF2 on vascular ECs in vascular dysfunction related diseases.

Objective To explore the influencing factors of compliance with new oral anticoagulants(NOACs)in elderly patients with nonvalvular atrial fibrillation(NVAF)and its correlation with social support.Methods The clinical data of 126 patients with NVAF treated by NOACs in Lu 'an People's Hospital of Anhui Province from January 2022 to January 2024 were selected by convenient sampling method.General data of patients were collected.According to Morisky medication compliance scale,patients were divided into two groups:the good compliance group(n=74)and the poor compliance group(n=52).Social support was investigated by Social Support Rating Scale(SSRS).General data of patients were compared between the good compliance group and the poor compliance group.The relationship between medication compliance and social support was analyzed by Pearson correlation.Multivariate Logistic regression was used to analyze the influencing factors of patient medication compliance.Results There were statistically significant differences in educational level,the proportion of patients with anxiety and depression,CHA2DS2-VASc score,HAS-BLED score and SSRS score between the good compliance group and the poor compliance group(P＜0.05).The results of the correlation analysis showed that the medication compliance of patients was positively correlated with objective support(r=0.461),subjective support(r=0.432),support utilization(r=0.323)and SSRS score(r=0.596,P＜0.05).The results of multivariate Logistic regression analysis showed that low educational level,accompanied by anxiety,low HAS-BLED and SSRS scores were risk factors for poor medication compliance in elderly NVAF patients(P＜0.05).Conclusion Improving the social support level of elderly patients with NVAF is conducive to the formation of good medication compliance.

Aim To construct a model of vascular endothelial cell(EC)-specific gene knockout mice of tumor necrosis factor receptor-associated factor 2(Traf2)by using Cre-loxP technolo-gy,thus to provide basis for the research of vascular dysfunction-related diseases related to the regulation of vascular EC activity through TRAF2.Methods The Cre-loxP system was used to construct a mouse model with EC-specific knockout of Traf2 gene(Traf2flox/flox Tek-iCre+).The mouse genotype was identi-fied through PCR and gel electrophoresis.Primary vascular ECs were isolated from the mice using flow cytometry.The knockout efficiency of Traf2 in vascular ECs was validated by Western blot and immunofluorescence.The pathological changes in blood ves-sels and major organs of the mice were examined using hematox-ylin-eosin(HE)staining.The tube-forming ability of primary vascular EC was assessed using Matrigel tube formation.Results The knockout mice met the required genetic criteria.Flow cy-tometry results demonstrated the successful isolation of primary vascular EC,and TRAF2 expression in vascular EC of knockout mice was significantly reduced(P＜0.01).Histological results showed that TRAF2 expression in the vessel of knockout mice decreased,and the morphology had no significant changes in their blood vessels and major organs.The Matrigel tube forma-tion demonstrated that the tube-forming ability of primary vascu-lar EC from knockout mice was reduced.Conclusion Traf2 specific knockout mouse model in vascular ECs is successfully constructed,providing a reliable animal model for research into the regulatory mechanisms of TRAF2 on vascular ECs in vascular dysfunction related diseases.

Objective To explore the influencing factors of compliance with new oral anticoagulants(NOACs)in elderly patients with nonvalvular atrial fibrillation(NVAF)and its correlation with social support.Methods The clinical data of 126 patients with NVAF treated by NOACs in Lu 'an People's Hospital of Anhui Province from January 2022 to January 2024 were selected by convenient sampling method.General data of patients were collected.According to Morisky medication compliance scale,patients were divided into two groups:the good compliance group(n=74)and the poor compliance group(n=52).Social support was investigated by Social Support Rating Scale(SSRS).General data of patients were compared between the good compliance group and the poor compliance group.The relationship between medication compliance and social support was analyzed by Pearson correlation.Multivariate Logistic regression was used to analyze the influencing factors of patient medication compliance.Results There were statistically significant differences in educational level,the proportion of patients with anxiety and depression,CHA2DS2-VASc score,HAS-BLED score and SSRS score between the good compliance group and the poor compliance group(P＜0.05).The results of the correlation analysis showed that the medication compliance of patients was positively correlated with objective support(r=0.461),subjective support(r=0.432),support utilization(r=0.323)and SSRS score(r=0.596,P＜0.05).The results of multivariate Logistic regression analysis showed that low educational level,accompanied by anxiety,low HAS-BLED and SSRS scores were risk factors for poor medication compliance in elderly NVAF patients(P＜0.05).Conclusion Improving the social support level of elderly patients with NVAF is conducive to the formation of good medication compliance.

Objective:To understand the serological characteristics of irregular antibodies in pregnant women and explore their clinical significance.Methods:From January 2017 to March 2022,151 471 pregnant women in Women and Children's Hospital of Chongqing Medical University were enrolled in this study,microcolumn gel card test was used for irregular antibody screening,and antibody specificity identification was further performed in some antibody-positive subjects.Results:The positive rate of irregular antibody screening in the enrolled pregnant women was 0.91％(1 375/151 471),0.23％(355/151 471)was detected in the first trimester,0.05％(71/151 471)in the second trimester,and 0.63％(949/151 471)in the third trimester.The positive rate of irregular antibody screening in the third trimester was significantly higher than that in the first and second trimester,and a significant increase in the number of positive cases was found in the third trimester than that in the second trimester.The analysis of agglutination intensity of 1 375 irregular antibody screening positive results showed that the weakly positive agglutination intensity accounted for 50.11％(689/1 375),which was the highest,the suspicious positive was 18.69％(257/1 375),and the positive was 31.20％(429/1 375).The significant difference in distribution of agglutination intensity was not observed between the first trimester group and the second trimester group,however,in the third trimester,the proportion of suspicious positive and weakly positive was lower than the first trimester,while,the proportion of positive was higher than the first trimester,and the difference was statistically significant(P＜0.001).Among the irregular antibody screening positive pregnant women,the proportion of pregnant women with pregnancy number ≥ 2 was significantly higher than that with pregnancy≤1.Among 60 pregnant women who underwent antibody identification,the distributions of the antibodies were as follows:Rh blood group system accounted for 23.33％(14/60),Lewis system 43.33％(26/60),Kidd system 3.33％(2/60),MNS system 16.67％(10/60),P1PK system 1.67％(1/60),autoantibodies 1.67％(1/60),and 4 cases was unable to identify(6.67％,4/60).Among specific antibodies,the anti-Lea was the most common(30.00％),followed by anti-E(16.67％)and anti-M(16.67％).Conclusion:The differences of irregular antibody serological characteristics exist in pregnant women from different regions with different genetic backgrounds,understanding the characteristics of irregular antibody in local pregnant women is of great significance for ensuring transfusion safety in pregnant women and preventing hemolytic disease of newborn.

Objective: To analyze the difference in blood uric acid levels between patients with polycystic ovary syndrome (PCOS) and healthy women of childbearing age, and to investigate the correlation between body composition and blood uric acid levels. Methods: A total of 153 eligible childbearing age patients with PCOS treated at Tianjin Medical University General Hospital from January 2018 to March 2022 were selected, and 153 healthy women with normal menstruation were selected as the control group. Fasting blood uric acid levels were measured by venous blood test, and body composition was measured by a body composition analyzer. Group comparisons were made to analyze the correlation between body composition and blood uric acid levels. Results: The incidence of hyperuricemia was higher in patients with PCOS than that in the control group [30.1% (46/153) vs 2.0% (3/153)], with a statistically significant difference (χ²=44.429, P<0.001). Blood uric acid level was also significantly higher in patients with PCOS than that in the control group [(371±98) vs (265±67) μmol/L; t=11.170, P<0.001]. Among PCOS patients, there were statistically significant differences in weight, body mass index (BMI), body fat mass, skeletal muscle mass, percent body fat, lean body weight, fat mass/lean body weight, percent skeletal muscle, and visceral fat level between the hyperuricemia group and the normal blood uric acid group (all P<0.001), but no significant difference was observed in waist-hip ratio (P=0.348). The following body composition indicators: weight, BMI, waist-hip ratio, body fat mass, skeletal muscle mass, percent body fat, visceral fat level, lean body weight, and fat mass/lean body weight in all subjects, the PCOS patients and the control group, were positively correlated with blood uric acid levels (all P<0.01). The blood uric acid level in PCOS obese patients was higher than that in non-obese PCOS patients, and the difference was statistically significant [(425±83) vs (336±91) μmol/L; t=6.133, P<0.001]. The blood uric acid level in central obesity PCOS patients was also higher than that in non-central obesity PCOS patients [(385±95) vs (299±79) μmol/L], the difference was statistically significant (t=4.261, P<0.001). The blood uric acid level in normal-weight obese PCOS patients was higher than that in normal-weight non-obese PCOS patients [(333±73) vs (277±54) μmol/L], and the difference was statistically significant (t=2.848, P=0.006). Blood uric acid levels in normal-weight [(315±74) vs (255±67) μmol/L], overweight [(362±102) vs (276±57) μmol/L], and obese PCOS patients [(425±83) vs (303±74) μmol/L] were all higher than those in the corresponding control groups, with statistically significant differences (all P<0.001). Conclusions: PCOS patients have a higher incidence of hyperuricemia than healthy women of childbearing age. Blood uric acid levels are closely correlated with body composition indicators, such as weight, BMI, waist-hip ratio, body fat mass, skeletal muscle mass, percent body fat, and visceral fat level. Body composition analysis of women with PCOS could help identify potentially obese people more accurately and carry out individualized treatment, thereby reducing the risk of metabolic abnormalities.
Humans ; Female ; Polycystic Ovary Syndrome/complications* ; Uric Acid ; Hyperuricemia/complications* ; Insulin ; Body Composition/physiology* ; Obesity/complications* ; Body Mass Index

Objective:To examine the significance of serum Endocan levels in evaluating the severity of disease and predicting the prognosis for elderly patients with multiple organ dysfunction syndrome(MODS).Methods:Seventy-five elderly patients from the Department of Geriatric ICU at the First Affiliated Hospital of Nanjing Medical University were selected and divided into two groups: the non-MODS group and the MODS group.This division was based on the levels of acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)scores and MODS scores within 24 hours of admission.Additionally, 40 elderly individuals undergoing physical examination were included as a control group.Venous blood samples were collected from all participants to detect the levels of Endocan using ELISA.The patients were further categorized into the survival group and the death group based on their clinical outcome within 28 days.The Endocan levels in all groups were compared, and receiver operating characteristic(ROC)curves were generated to assess the value of Endocan in determining the severity of the disease and predicting prognosis in elderly MODS patients.Results:The levels of Endocan were found to be higher in the MODS group compared to the non-MODS and control groups[(622.3±149.2)ng/L vs.(433.1±189.7)ng/L, P<0.001]. Furthermore, patients with higher APACHE Ⅱ and MODS scores exhibited higher levels of Endocan.The area under the ROC curve(AUC)for Endocan, MODS score, and Endocan+ MODS score were 0.791, 0.806, and 0.820, respectively( P>0.05). Similarly, the AUC for Endocan, APACHE Ⅱ score, and Endocan+ APACHE Ⅱ score were 0.763, 0.799, and 0.803, respectively( P>0.05). Interestingly, the survival group had lower levels of Endocan compared to the death group[(444.6±193.6)ng/L vs.(618.2±149.5)ng/L, P<0.001]. Conclusions:Endocan can be utilized as a noteworthy indicator, which is associated with the seriousness of illness and the prognosis for elderly patients with multiple organ dysfunction syndrome(MODS).

Objective: To produce latent membrane protein 2A (LMP2A) chimeric antigen receptor (CAR)-T cells and detect the lethal effect of LMP2A CAR-T cells on nasopharyngeal carcinoma (NPC) cells. Methods: The study was conducted from September 2016 to December 2017.Genetic engineering technology was used to construct anti-LMP2A CAR lentiviral expression vector and sequencing was identified. The expression of anti-LMP2A CAR in the 293T cells was confirmed by western blot. CCK8 assay was used to evaluate the cytotoxicity of LMP2A CAR-T cells to NPC cells. ELISA assay was performed to test IL-2 and IFN-γ releasing of activated LMP2A CAR-T cells. The inhibition effect of LMP2A CAR-T cells on NPC xenograft tumor was observed in vivo. Statistical analysis was performed by statistical software SPSS 21.0. Results: The results of PCR and sequencing showed that anti-LMP2A CAR lentiviral expression vector was constructed successfully. The result of western blot indicated the expression of anti-LMP2A CAR in the 293T cells effectively. The results of CCK-8 assay showed that the killing activities of LMP2A CAR-T cells to LV-LMP2A-CNE1 cells were (72.11±9.75)%, (54.65 ±5.42)% and (36.68±3.80)% at 20∶1, 10∶1 and 5∶1 ratio of effective cells to target cells, and had a statistical difference compared to CD19 CAR-T cells and T cells (P<0.05). There was no significant difference in the killing activities of LMP2A CAR-T cells to CNE1 cells compared with CD19 CAR-T cells and T cells. The results of ELISA showed that the content of IL-2 and IFN-γ in the co-culture supernatant of LMP2A CAR-T cells and LV-LMP2A-CNE1 cells was significantly higher than that of LMP2A CAR-T cells and CNE1 cells which had statistical difference (P<0.05); In vivo experiment, the volume of LMP2A CAR-T cell group was (80.3±10.0) mm³ which was significantly lower than that of the control groups, and the difference was statistically significant (P<0.05). Conclusion LMP2A CAR-T cells are successfully prepared and have an obvious targeting cytotoxicity on LMP2A-positive NPC cells.

Objective To evaluate the quality of blood after marine storage and transportation,to establish a quantitative energy metabolism platform supported by extracellular flux(XF)technique to determine the glycolysis level of red blood cells(RBC), and to introduce the "functional dose", as a valuable supplement, into the quality evaluation of RBCs after marine storage and transportation.Methods The extracellular acidification rate(ECAR)of erythrocyte glycolysis was detected by XF.The levels of glycolysis, ATP contents and free hemoglobin of RBCs were detected before and after marine navigation.RBC suspensions stored at 4℃in a blood bank on land served as control.Results The results showed that the glycolysis level of RBCs was detected by XF.Indicators of blood quality and energy metabolism of RBCs were detected in the twice marine storage and transportation.Blood samples transported around the Gulf of Bohai had no significant change,indicating that the samples were well preserved.The energy indicators of blood samples navigated in the Pacific Ocean decreased significantly,and it was recommended that the samples be discarded.Conclusion XF technique has been applied to the measurement of erythrocyte glycolysis,enabling high-throughput and multi-indicator dynamic detection of viable cells.The energy metabolism of erythrocyte should be recommended as an important indicator for evaluating blood after marine storage and transportation.

Objective To investigate the application of electronic bronchoscopy in diagnosis of recurrent tracheoe-sophageal fistula of type Ⅲ esophageal atresia. Methods 5 patients were reviewed who were suspected postopera-tive tracheoesophageal fistula recurrence of type Ⅲ esophageal atresia and examined through electronic bron-choscopy combined with injecting Methylene blue into stomach tube from Jan 2010 to Aug 2014. Male to female ra-tio was 4:1. The age was 2~15 months, median age was 7.4 months. Results The 5 cases were found trachea mem-brane fistula by electronic bronchoscope, recurrent tracheoesophageal fistula was diagnosed with Methylene blue in-jected into stomach tube and overflowed from trachea membrane fistula. Examination time is 3~7 minutes, the aver-age is 4.2 minutes. The 5 cases were confirmed by operation. Conclusion Electronic bronchoscopy combined with injecting Methylene blue into stomach tube to diagnose recurrent tracheoesophageal fistula is safe and feasible, the time is short and the diagnosis rate is high.