Objective: To observe the changes of PD-1 expression, mRNA level and cytotoxic activity of CD19 CAR-T cells during the culture process of CAR-T cells. Methods: The peripheral blood T cells of 6 lymphoma patients with high expression of PD-1 and 6 healthy volunteers were the source of CAR-T cells. The expression of PD-1 was analyzed by flow cytometry. The mRNA level of PD-1 was analyzed by PCR. The cell proliferation was analyzed by CCK-8 assay. The cytotoxicity was analyzed by LDH assay. Results: ①The transfection efficiency of high PD-1 expression T cells and healthy volunteer T cells were as the same (P>0.05) . ②The cell proliferation capacity of CD19 CAR-T cells from high PD-1 expression T cells or healthy volunteer T cells, with or without PD-1 inhibitor were as the same (P>0.05) . ③The cytotoxicity to lymphoma cells of high PD-1 expression T cells and CAR-T cells were lower than that of these two T cells combined with PD-1 inhibitor and the CAR-T cells from healthy volunteer T cells (P<0.001) . There was no difference of the cytotoxicity between the CAR-T cells from high PD-1 expression T cells combined with PD-1 inhibitor and the CAR-T cells from healthy volunteer (P>0.05) . ④There was no difference of the expression of PD-1 in all CAR-T cell groups during the culture process (P>0.05) . There was no difference of mRNA level of PD-1 in all groups during the culture process (P>0.05) . ⑤The PD-1 expression of CAR-T cells increased by the time of culture after contacting with lymphoma cells (P<0.001) . The PD-1 inhibitors could antagonize this effect. There was no difference of mRNA level of PD-1 in all groups after contacting with lymphoma cells (P>0.05) . Conclusion The PD-1 expression of CAR-T cells from high PD-1 expression T cells increased by the time of culture after contacting with lymphoma cells. However, the mRNA level of PD-1 of all groups did not change, even if PD-1 inhibitor was applied.

Objective:To evaluate the diagnostic and follow-up value of ultrasonography for juvenile dermatomyositis (JDM).Methods:Ten children with newly diagnosed or relapsed JDM in Children′s Hospital, Capital Institute of Pediatrics from October 2021 to October 2022 and 15 healthy children were prospectively collected. The clinical data of JDM children were collected, and the muscle ultrasound was performed at the first diagnosis, 3, 6, 12 and 18 months after diagnosis. The ultrasound parameters including quantitative muscle echogenicity (MEI), fascia thickness (FT), microvascular imaging (MVI) distribution, blood resistance index (RI), the changes in the characteristics of the sonogram were observed. The correlation between ultrasound data and pediatric myositis assessment scale (CMAS) score, creatine kinase (CK) were analyzed during the follow-up. Results of muscle ultrasound in children with first active JDM and normal children was compared.Results:After 18 months of treatment, MEI decreased significantly (75.62±4.32 vs 41.81±12.50, P<0.01), FT decreased[(0.27±0.06)cm vs (0.20±0.05)cm, P<0.01], and MVI distribution decreased[0 vs 7(70%), P<0.01] in 10 children with JDM.Spearman correlation analysis showed that MEI, FT, and MVI distribution were negatively correlated with CMAS score ( rs=-0.771, -0.443, -0.686; all P<0.05), while increased MEI and MVI distribution were positively correlated with CK ( rs=0.463, 0.464; all P<0.05). MEI returned to normal in 3 cases, FT didn′t completely recover, and the soft tissue calcification appeared. MEI, FI, and MVI distribution were significantly higher and RI was lower in children with JDM than in normal children (all P<0.01). Conclusions:Muscle ultrasound is suitable for the long-term dynamic detection of JDM, and the activity status of JDM disease can be judged by MEI, FT and MVI.

Objective To observe the characteristics of infant biliary atresia(BA)missed diagnosis by ultrasound.Methods Data of 72 BA infants missed by ultrasound(false-negative,group A),72 BA infants accurately diagnosed by ultrasound(true-positive,group B)and 72 non BA infants(controls,group C)were retrospectively analyzed.Clinical and ultrasonic features were compared between each 2 groups.Logistic regression analysis was performed to screen the impact factors for diagnosing and missed-diagnosing of BA.Results There were significant differences of total bilirubin,direct bilirubin,gamma glutamyl transpeptidase(GGT)and glutamic-oxaloacetic transaminase(GOT)between group A and C,also of total bilirubin,direct bilirubin and GGT between group B and C(all P＜0.05).Significant differences of the incidences of small gallbladder,gallbladder wall rigidity,poor gallbladder contraction function and fibrous plaques in the hepatic hilum were found between group A and B,of undetected or cystic changes in the gallbladder,gallbladder wall rigidity,poor gallbladder contraction function,fibrous plaques in the hepatic hilum and hepatic hilum cysts were noticed between group A and C(all P＜0.05),as well as of undetected or cystic changes in the gallbladder,small gallbladder,gallbladder wall rigidity,gallbladder wall thickening,poor gallbladder contraction function and fibrous plaques in the hepatic hilum were detected between group B and C(all P＜0.05).There were significant differences of display rate of common bile duct between each 2 groups(all P＜0.05).The lower incidence of fibrous plaques in the hepatic hilum and poor gallbladder contraction function were an impact factors for missed diagnosis of BA(both P＜0.05),while higher GGT,higher incidence of fibrous plaques in the hepatic hilum and poor gallbladder contraction function were all impact factors for diagnosis of BA in group A(all P＜0.05).Conclusion Infant BA missed diagnosis by ultrasound had lower incidence of fibrous plaques in the hepatic hilum and poor gallbladder contractile function.Combining with clinical data was helpful for reducing ultrasonic missed diagnosis of BA in infants.

Objective:To elucidate the current proficiency level in diagnosing pediatric biliary atresia among Chinese sonographers and to identify contributing factors that may influence their diagnostic capabilities.Methods:A cross-sectional internet-based survey was conducted from November 6, 2021, to December 12, 2021. The online questionnaire was disseminated to a national cohort of sonographers via WeChat groups. The survey encompassed three domains: demographic data of the participants, clinical and ultrasonographic knowledge pertaining to biliary atresia, and the cognitive understanding of biliary atresia. Descriptive statistics were utilized for the evaluation of demographic information and sonographers' clinical and ultrasonographic comprehension concerning biliary atresia. A generalized linear model was employed to ascertain the determinants influencing sonographers' expertise in clinical management and ultrasonographic diagnosis of biliary atresia.Results:A total of 511 valid responses were collated. Approximately 64.77% (331/511) of sonographers indicated a lack of comprehensive understanding of the essential parameters for ultrasonographic diagnosis of biliary atresia. Regression analysis revealed that sonographers affiliated with lower-tier hospitals ( β=-0.344, P=0.002), non-pediatric institutions ( β=-0.747, P=0.004), and those with less than 15 years of professional experience ( β=0.952, P<0.001) exhibited diminished proficiency in diagnosing biliary atresia. Upon encountering suspected cases, a mere 5.48% (28/511) of sonographers expressed complete confidence in their diagnostic capabilities. Lower professional grade ( β=-0.572, P<0.001) and lack of prior experience with suspected cases of biliary atresia ( β=-0.693, P<0.001) contributed to reduced diagnostic self-assurance among sonographers. Regarding the clinical and ultrasonographic intricacies associated with biliary atresia, only 3.33% (17/511) of sonographers had a full understanding of the diagnostic significance of hepatic fibrous plaques on ultrasound for biliary atresia. Conclusion:The study underscores a pervasive inadequacy in the understanding and diagnostic confidence among sonographers in relation to pediatric biliary atresia. Various pragmatic factors, including hospital tier, years of experience, and professional ranking, exert a direct impact on the sonographers' diagnostic acumen. Given these findings, there is an exigent need for targeted educational initiatives to enhance the diagnostic proficiency in biliary atresia among sonographers.

Objective:To observe the local reactions and efficacy of CD19 CAR-T therapy in recurrence/refractory B-cell non-Hodgkin's lymphoma (R/R NHL) patients with >7.5 cm lesions.Methods:32 R/R NHL patients with >7.5 cm lesions were enrolled and injected with CD19 CAR-T cells. Flow cytometry was used to detect and observe the amplification of CD19 CAR-T cells in vivo. Enzyme-linked immunosorbent assay (ELISA) was used to detect cytokines in peripheral blood of patients. The side effects of CD19 CAR-T cell therapy included systemic side effects and local reactions of tumor. The local side effects were observed by Ultrasound, Computed tomography and Magnetic resonance imaging. Treatment options included glucocorticoid, interleukin-6 antibody and drainage of exudate. Overall response rate (ORR) and overall survival rate (OS) were observed.Results:①Among the 32 patients, CR (40.63%) , PR (31.25%) and ORR (71.88%) were 13, 10 and 23, respectively. ②In all 23 patients received ORR, 13 patients had grade 1-2 CRS, while 10 patients had grade 3-4 CRS. All the 9 patients in the SD+PD group had grade 1-2 CRS ( P=0.030) . ③A total of 15 patients with tumor local reactions, included 9 patients with CR, 5 patients with PR and 1 patient with SD. The local reactions of the tumor included that the diameter of the superficial lesions increased with redness, swelling and heat pain. The deep lesions presented abdominal pain, abdominal distension, suffocation and local pain, and burning of the tumor. The deep lesions were enlarged or accompanied by local edema. The local exudative lesions were found in the abdominal cavity and pleural cavity. ④ Peak proportion of CD19 CAR-T cells in ORR group was higher than that of in SD+PD group[16.8% (5.3%-48.2%) vs 2.9% (1.5%-5.7%) , z=-4.297, P<0.001]. The peak proportion of CD19 CAR-T cells in ORR group with local reactions was higher than that of in patients without local reactions [22.2% (10.5%-48.2%) vs 12.6% (5.3%-21.6%) , z=-3.213, P=0.001]. The peak proportion of CD19 CAR-T cells in multiple lesion group was higher than that of in single lesion group [35.8% (1.5%-48.2%) vs 16.8% (10.5%-18.5%) , z=-2.023, P=0.040]. ⑤Occurrence of local reactions and tumor shrinkage time were both delayed compared with systemic side effects. ⑥In the ORR group, the OS of patients with tumor local reactions was longer than that of patients without tumor local reactions, but there was no difference in the two groups (75% vs 34.6%, P=0.169) . Conclusions:CD19 CAR-T cell therapy in R/R NHL patients with >7.5 cm lesions might cause tumor local reactions later than systemic side effects.Clinicaltrial::ChiCTR1800018059