OBJECTIVE: To explore the clinical phenotype and genetic characteristics of a pediatric child with RELA-associated autoinflammatory disease (RAID) caused by a RELA gene variant, and to review the reported cases in the literature. METHODS: A pediatric child with RAID who presented with recurrent fever, vomiting, and oral ulcers for over 5 years was selected as the study subject. The child visited the Women and Children's Hospital of Ningbo University in August 2023. Clinical data were collected, and peripheral blood samples were obtained from the child and his family members for whole-exome sequencing (WES) and Sanger sequencing to identify and validate candidate variants. The pathogenicity of the variants was analyzed accordingly. Using the keywords "RELA" "NF-κB" "autoinflammatory disease" "tofacitinib" "sulfasalazine" a literature search was conducted in the China National Knowledge Infrastructure, Wanfang Data Knowledge Service Platform, and PubMed from January 1, 2000 to December 13, 2023. This study was approved by the Medical Ethics Committee of the Women and Children's Hospital of Ningbo University (Ethics No. EC2020-048). RESULTS: The child primarily manifested with recurrent fever, vomiting, and oral ulcers. WES identified a heterozygous nonsense variant c.985C>T (p.Arg329Ter) in the RELA gene, which was inherited from the mother. According to the American College of Medical Genetics and Genomics (ACMG) Standards and Guidelines for the Interpretation of Sequence Variants and the Clinical Genome Resource (ClinGen) recommendations for PVS1, this variant was classified as pathogenic (PVS1+PM2_Supporting+PP4). Despite treatment with adalimumab and tocilizumab, the child's symptoms persisted. Switching to tofacitinib improved oral ulcers, but fever and vomiting continued. The addition of thalidomide significantly alleviated fever and vomiting, and the patient's growth and development remained normal. A literature review identified 14 unrelated RAID families, including a total of 35 cases (including the present child). The main clinical features were recurrent oral ulcers, genital ulcers, skin problems, fever, diarrhea, abdominal pain, and vomiting. CONCLUSION The nonsense variant c.985C>T (p.Arg329Ter) in the RELA gene is likely the genetic cause of the child's recurrent fever, vomiting, and oral ulcers. WES is valuable for timely diagnosis of RAID and provides a basis for clinical treatment strategies.
Humans ; Male ; Transcription Factor RelA/genetics* ; Female ; Hereditary Autoinflammatory Diseases/genetics* ; Child ; Pedigree ; Exome Sequencing

Objective:To explore the clinical phenotype and genetic characteristics of a pediatric child with RELA-associated autoinflammatory disease (RAID) caused by a RELA gene variant, and to review the reported cases in the literature. Methods:A pediatric child with RAID who presented with recurrent fever, vomiting, and oral ulcers for over 5 years was selected as the study subject. The child visited the Women and Children′s Hospital of Ningbo University in August 2023. Clinical data were collected, and peripheral blood samples were obtained from the child and his family members for whole exome sequencing (WES) and Sanger sequencing to identify and validate candidate variants. The pathogenicity of the variants was analyzed accordingly. Using the keywords " RELA" " NF-κB" " autoinflammatory disease" " tofacitinib" " sulfasalazine" a literature search was conducted in the China National Knowledge Infrastructure, Wanfang Data Knowledge Service Platform, and PubMed from January 1, 2000 to December 13, 2023. This study was approved by the Medical Ethics Committee of the Women and Children′s Hospital of Ningbo University (Ethics No. EC2020-048).Results:① The child primarily manifested with recurrent fever, vomiting, and oral ulcers. ② WES identified a heterozygous nonsense variant c. 985C>T (p.Arg329Ter) in the RELA gene, which was inherited from the mother. According to the American College of Medical Genetics and Genomics (ACMG) Standards and Guidelines for the Interpretation of Sequence Variants and the Clinical Genome Resource (ClinGen) recommendations for PVS1, this variant was classified as pathogenic (PVS1+ PM2_Supporting+ PP4). ③ Despite treatment with adalimumab and tocilizumab, the child′s symptoms persisted. Switching to tofacitinib improved oral ulcers, but fever and vomiting continued. The addition of thalidomide significantly alleviated fever and vomiting, and the patient′s growth and development remained normal. ④ A literature review identified 14 unrelated RAID families, including a total of 35 cases (including the present child). The main clinical features were recurrent oral ulcers, genital ulcers, skin problems, fever, diarrhea, abdominal pain, and vomiting. Conclusion:The nonsense variant c. 985C>T (p.Arg329Ter) in the RELA gene is likely the genetic cause of the child′s recurrent fever, vomiting, and oral ulcers. WES is valuable for timely diagnosis of RAID and provides a basis for clinical treatment strategies.

Objective:To explore the effects of smoking on peripheral regulatory T cells (Tregs), transforming growth factor beta1 (TGF-β 1) and interleukin-10 (IL-10) in elderly patients with non-small cell lung cancer (NSCLC). Methods:This was a cross-sectional study. A total of 43 elderly patients (≥60 years old) who were hospitalized in the Department of Geriatrics Medicine, Guangzhou First People′s Hospital from January 2018 to December 2024 and were newly diagnosed with NSCLC were recruited. According to smoking history, patients were divided into non-smoking group (15 cases), low smoking group (13 cases, smoking index<400) and high smoking group (15 cases, smoking index≥400). Venous blood samples were collected from participants, plasma and cells were separated. Flow cytometry was used to measure the proportions of Tregs and the expression of forkhead box P3 (Foxp3) in peripheral blood. Plasma levels of TGF-β 1 and IL-10 were measured by enzyme-linked immunosorbent assay. The effects of smoking on peripheral Tregs, TGF-β 1, and IL-10 in elderly patients with NSCLC were analyzed. Data were analyzed by one-way ANOVA, rank-sum test, and Fisher′s exact test. Results:The proportions of Tregs in non-smoking group, low smoking group and high smoking group were 2.50% (2.32%, 2.81%), 2.83% (2.48%, 3.72%), and 3.01% (2.37%, 3.73%), respectively, and there were no statistically significant differences among the three groups ( H=3.845, P>0.05). The proportions of Foxp3 +Tregs were (3.72±0.84)%, (4.64±1.10)%, and (4.68±1.27%), respectively. The mean fluorescence intensities (MFI) of Foxp3 were 123.0 (108.0, 128.0), 131.0 (123.5, 350.0), and 222.0 (141.0, 311.0), respectively. Both the proportions of Foxp3 +Tregs and the MFI of Foxp3 were higher in low smoking group and high smoking group than those in non-smoking group (all P<0.05). However, there were no significant differences between low smoking group and high smoking group (all P>0.05). The concentrations of IL-10 were 2.27 (1.42, 3.95), 3.42 (2.30, 5.08), and 3.26 (2.35, 6.28) ng/L, respectively. There were no statistically significant differences among the three groups ( H=2.930, P>0.05). The concentrations of TGF-β 1 were (10.72±9.37), (13.46±10.39), and (25.28±16.67) ng/ml, respectively. The concentration of TGF-β 1 in high smoking group was higher than that in non-smoking group and low smoking group (all P<0.05). However, there was no statistically significant difference between low smoking group and non-smoking group ( P>0.05). Conclusions:Smoking intensity may be correlated with the immunosuppressive function of Tregs in elderly patients with NSCLC. Higher smoking levels are associated with increased Foxp3 expression in Tregs and elevated plasma levels of TGF-β 1, potentially enhancing the immunosuppressive function of Tregs.

Objective:To explore the clinical phenotype and genetic characteristics of a pediatric child with RELA-associated autoinflammatory disease (RAID) caused by a RELA gene variant, and to review the reported cases in the literature. Methods:A pediatric child with RAID who presented with recurrent fever, vomiting, and oral ulcers for over 5 years was selected as the study subject. The child visited the Women and Children′s Hospital of Ningbo University in August 2023. Clinical data were collected, and peripheral blood samples were obtained from the child and his family members for whole exome sequencing (WES) and Sanger sequencing to identify and validate candidate variants. The pathogenicity of the variants was analyzed accordingly. Using the keywords " RELA" " NF-κB" " autoinflammatory disease" " tofacitinib" " sulfasalazine" a literature search was conducted in the China National Knowledge Infrastructure, Wanfang Data Knowledge Service Platform, and PubMed from January 1, 2000 to December 13, 2023. This study was approved by the Medical Ethics Committee of the Women and Children′s Hospital of Ningbo University (Ethics No. EC2020-048).Results:① The child primarily manifested with recurrent fever, vomiting, and oral ulcers. ② WES identified a heterozygous nonsense variant c. 985C>T (p.Arg329Ter) in the RELA gene, which was inherited from the mother. According to the American College of Medical Genetics and Genomics (ACMG) Standards and Guidelines for the Interpretation of Sequence Variants and the Clinical Genome Resource (ClinGen) recommendations for PVS1, this variant was classified as pathogenic (PVS1+ PM2_Supporting+ PP4). ③ Despite treatment with adalimumab and tocilizumab, the child′s symptoms persisted. Switching to tofacitinib improved oral ulcers, but fever and vomiting continued. The addition of thalidomide significantly alleviated fever and vomiting, and the patient′s growth and development remained normal. ④ A literature review identified 14 unrelated RAID families, including a total of 35 cases (including the present child). The main clinical features were recurrent oral ulcers, genital ulcers, skin problems, fever, diarrhea, abdominal pain, and vomiting. Conclusion:The nonsense variant c. 985C>T (p.Arg329Ter) in the RELA gene is likely the genetic cause of the child′s recurrent fever, vomiting, and oral ulcers. WES is valuable for timely diagnosis of RAID and provides a basis for clinical treatment strategies.

Objective:To explore the effects of smoking on peripheral regulatory T cells (Tregs), transforming growth factor beta1 (TGF-β 1) and interleukin-10 (IL-10) in elderly patients with non-small cell lung cancer (NSCLC). Methods:This was a cross-sectional study. A total of 43 elderly patients (≥60 years old) who were hospitalized in the Department of Geriatrics Medicine, Guangzhou First People′s Hospital from January 2018 to December 2024 and were newly diagnosed with NSCLC were recruited. According to smoking history, patients were divided into non-smoking group (15 cases), low smoking group (13 cases, smoking index<400) and high smoking group (15 cases, smoking index≥400). Venous blood samples were collected from participants, plasma and cells were separated. Flow cytometry was used to measure the proportions of Tregs and the expression of forkhead box P3 (Foxp3) in peripheral blood. Plasma levels of TGF-β 1 and IL-10 were measured by enzyme-linked immunosorbent assay. The effects of smoking on peripheral Tregs, TGF-β 1, and IL-10 in elderly patients with NSCLC were analyzed. Data were analyzed by one-way ANOVA, rank-sum test, and Fisher′s exact test. Results:The proportions of Tregs in non-smoking group, low smoking group and high smoking group were 2.50% (2.32%, 2.81%), 2.83% (2.48%, 3.72%), and 3.01% (2.37%, 3.73%), respectively, and there were no statistically significant differences among the three groups ( H=3.845, P>0.05). The proportions of Foxp3 +Tregs were (3.72±0.84)%, (4.64±1.10)%, and (4.68±1.27%), respectively. The mean fluorescence intensities (MFI) of Foxp3 were 123.0 (108.0, 128.0), 131.0 (123.5, 350.0), and 222.0 (141.0, 311.0), respectively. Both the proportions of Foxp3 +Tregs and the MFI of Foxp3 were higher in low smoking group and high smoking group than those in non-smoking group (all P<0.05). However, there were no significant differences between low smoking group and high smoking group (all P>0.05). The concentrations of IL-10 were 2.27 (1.42, 3.95), 3.42 (2.30, 5.08), and 3.26 (2.35, 6.28) ng/L, respectively. There were no statistically significant differences among the three groups ( H=2.930, P>0.05). The concentrations of TGF-β 1 were (10.72±9.37), (13.46±10.39), and (25.28±16.67) ng/ml, respectively. The concentration of TGF-β 1 in high smoking group was higher than that in non-smoking group and low smoking group (all P<0.05). However, there was no statistically significant difference between low smoking group and non-smoking group ( P>0.05). Conclusions:Smoking intensity may be correlated with the immunosuppressive function of Tregs in elderly patients with NSCLC. Higher smoking levels are associated with increased Foxp3 expression in Tregs and elevated plasma levels of TGF-β 1, potentially enhancing the immunosuppressive function of Tregs.

Objective:To explore the genetic etiology for a child with Progressive external ophthalmoplegia with mitochondrial DNA deletions, autosomal dominant 6 (PEOA6).Methods:A child who had attended the Women and Children′s Hospital Affiliated to Ningbo University on 7 August, 2023 was selected as the study subject. Clinical data of the child were analyzed retrospectively. The child and her parents were subjected to whole exome sequencing (WES), and candidate variant was verified by Sanger sequencing and bioinformatic analysis. This study was approved by Medical Ethics Committee of the Women and Children′s Hospital Affiliated to Ningbo University (Ethics No. EC2020-048).Results:The child, a 7-year-old female, had presented with limb muscle pain, amyosthenia, significantly increased creatine kinase, congenital diaphragmatic hernia and recurrent respiratory tract infections. WES revealed that she has harbored a heterozygous c. 1590G>C (p.L530F) variant of the DNA2 gene, which was verified to have a de novo origin by Sanger sequencing. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c.1590G>C was rated as a likely pathogenic variant (PS2+ PM2_Supporting+ PP3). Conclusion:The c.1590G>C (p.L530F) variant of the DNA2 gene probably underlay the PEOA6 in this child.

Objective:To evaluate the preoperative localization value of endoscopic ultrasound guided fine needle tattooing (EUS-FNT) for laparoscopic distal pancreatectomy in pancreatic lesions with a maximum diameter ≤3 cm.Methods:From November 2017 to October 2022, at the Second Affiliated Hospital of Soochow University, the data of patients with pancreatic lesions ≤3 cm who underwent laparoscopic distal pancreatectomy were retrospectively analyzed. Eight patients who underwent EUS-FNT assisted laparoscopic distal pancreatectomy were included in the fine needle tattooing (FNT) combined laparoscopic group. And 14 patients who underwent simple laparoscopic distal pancreatectomy were taken as the simple laparoscopic group. The success rate and complications of EUS-FNT were observed. The differences in operation time, surgery-related complications and complete resection rate of lesions between the two groups were compared. Mann-Whitney U test and descriptive analysis were used for statistical analysis. Results:In the FNT combined laparoscopic group, the lesions of 4 cases were located in the pancreatic body and 4 cases in the pancreatic tail. In the simple laparoscopic group, the lesions of 4 cases were located in the pancreatic body and 10 cases in the pancreatic tail. There was a significant difference in lesion size between the two groups (14.5 mm (10.8 mm, 16.5 mm) vs. 27.0 mm (23.5 mm, 30.0 mm), Z=-3.09, P=0.001). In the FNT combined laparoscopic group, EUS-FNT was successfully performed in all 8 patients. The average time of laparoscopy after EUS-FNT was (98.4±8.8) min. The marks were clearly visible under the laparoscopic field of view, and no complications such as abdominal hemorrhage and hematoma were observed. Laparoscopic pancreaticocaudectomy was performed in 5 cases and pancreaticocaudectomy plus splenectomy in 3 cases. The median operation time was 192.5 min (176.3 min, 203.8 min). The amount of intraoperative bleeding was large in 2 patients and blood transfusion was needed. The lesions were one-time completely resected in all 8 patients. The postoperative pathology were 6 cases of pancreatic neuroendocrine neoplasm, 1 case of intraductal papillary mucinous neoplasm (IPMN), and 1 case of solid pseudopapilloma. In the simple laparoscopic group, laparoscopic pancreaticocaudectomy was performed in 2 cases and pancreaticocaudectomy plus splenectomy in 12 cases. The median operation time was 202.5 min (192.8 min, 235.0 min), which was longer than that of FNT combined laparoscopic group, but the difference was not statistically significant ( P>0.05). The amount of intraoperative bleeding was large in 2 patients and blood transfusion was needed. In 1 patient with pancreatic body lesions, no lesion was found in the specimen examination after the first pancreatectomy, and the lesions were completely resected after the second partial pancreatectomy. Active abdominal hemorrhage occurred in 1 patient on the second day after operation, and underwent interventional embolization for hemostasis. Two weeks after surgery, 1 patient was found to have a encapsulated fluid with a long diameter of 6 cm around the pancreas by computed tomography re-examination 2 weeks after surgery. The postoperative pathology were 5 cases of pancreatic neuroendocrine neoplasm, 2 cases of IPMN, 1 case of solid pseudopapilloma, 1 case of pancreatic cyst with glandular low-grade intraepithelial neoplasia, 1 case of ectopic spleen, and 4 cases of pancreatic ductal adenocarcinoma. Conclusion:EUS-FNT can effectively localize small pancreatic lesions before laparoscopic distal pancreatectomy, shorten the operation time and improve the complete resection rate under laparoscopy.

New immune checkpoint targets such as lymphocyte activation gene-3, T-cell immunoglobulin mucin-3, T cell immunoglobulin and ITIM domain, V-domain immunoglobulin suppressor of T cell activation, B7 Homolog 3, B and T lymphocyte attenuator play an important role in tumor treatment. Blocking them can delay tumor growth, enhance the anti-tumor effect of immune cells, and improve the survival rate of the host. Although these new immune checkpoints are in the early stages of research, with the continuous development of science and technology, they will play an increasingly important role in tumor immunotherapy.

Objective To explore the correlation between the distribution of uric acid (UA) level and the biochemical indicators which reflect the degree of organ lesions among hospitalized children.Methods Patients who were hospitalized to the Department of Pediatrics and received the blood UA test from June 2012 to October 2013 were included,23 217 cases in total.The biochemical analyzer-Japan's Olympus AU 2700 was used to detect blood biochemistry; and uricase-peroxidase coupling method was used to detect blood UA.Among these patients,2 099 cases whose UA level exceeded the normal level.Then the patients' gender,age,primary diagnosis and UA level were recorded; and the distribution of their UA level was described.The Chi-square or Fisher test was used to assess the incidence rate.At the same time,each blood biochemical indicators of the patients with high UA level were recorded ; and the relationships between the blood UA of the 1 650 patients with complete records and each blood biochemical indicator were analyzed with Pearson correlation analysis.Results The incidence rate of hyperuricemia among the hospitalized children was 9.04％(2 099/23 217).In particular,the incidence rate among boys and girls was 6.5t％(890/13 657) and 12.65％ (1 209/9 560) respectively (x2 =256.9,P＜0.05).The incidence rate of hyperuricemiin different diseases was as follows:in the critical illness 36.93％ (113/306),neonatal disease 20.34％ (922/4 533),urinary system diseases 12.08％ (47/389),circulatory system diseases 11.67％ (21/180),nervous system diseases 11.05％(112/1 014),digestive diseases 10.50％(190/1 810),infectious diseases 10.18％(120/1 179),blood diseases 7.58％ (55/726),endocrine system diseases 5.74％ (17/296),autoimmune diseases 4.24％ (48/1 131),respiratory diseases 3.90％ (454/11 653) respectively (x2=1423.0,P＜0.05).The incidence of hyperuricemia at younger than one month was 18.31％(929/5 075),younger than one year old was 4.22％ (359/8 501),younger than six years old was 10.68％(600/5 618),younger than 15 years old was 5.24％ (211/4 023) respectively (x2=858.5,P＜0.05).Blood UA was positively correlated to urea nitrogen,creatinine,lacticdehydrogenase,α-hydroxy-butyrate dehydrogenase,creatine kinase and creatine kinase-MB (r=0.426,0.44,0.324,0.367,0.413,0.431,P＜0.05).Blood UA was not correlated to fructosamine,blood glucose,triglycerides,total cholesterol,low-density lipoprotein and high density lipoprotein.Conclusion The incidence of hyper-uricemia among hospitalized children is high; and the incidence among children with severe diseases and newborn babies is high; followed by in children with urinary system and circulatory system diseases.The blood UA level is closely related to the blood biochemical indicators which reflect the lesions of heart and kidney.

Objective To observe the levels of N-terminal pro-brain natriuretic peptide (NT-proBNP),high density lipoprotein cholesterol (HDL-C) in predicting cardiovascular damage in Kawasaki disease (KD).Methods Enzyme-linked fluorescence analysis (ELFA) technique was used to measure serum NT-proBNP levels in 406 KD patients [including 104 cases of incomplete Kawasaki disease (IKD)] at the acute phase,the convalescent stage,at the same time,the blood HDL-C,Albumin,globulin,alanine aminotransferase (ALT),C-reactive protein (CRP),red blood cell sedimentation rate (ESR),blood white cell count (WBC),hemoglobin (Hb),blood platelet count (PLT) level were tested.According to the results of echocardiography,electrocardiogram,myocardial enzymes in KD,patients were divided into two groups:two hundred and twenty-two with cardiovascular damage and 184 without cardiovascular damage group.The age,gender,fever,the first application of the intravenous gamma globulin,laboratory markers of clinical features observed above the detection levels were compared,and these parameters for each group were compared using t test or analysis of variance,the single factor which was statistically significant were received operating characteristic curve (ROC) analysis.Then the cardiovascular damage group was divided into echocardiography abnormal group and echocardiography normal group,and t test was adopted to compare the clinical parameters of the two groups.Results ① The age,lip and oral changes,the first application of intravenous gamma globulin,blood NT-proBNP,HDL-C,albumn and ALT was significantly different between the cardiovascular damage group and non cardiovascular damage group (t/x22=4.989,4.721,6.212,18.834,12.788,4.851,4.541,All P＜0.05).② When the blood NT-pro-BNP was higher than 786.5 ng/L,its sensitivity and specificity for differentiating KD with cardiovascular damage was 86.5％ and 84.8％,respectively.When the blood HDL-C was lower than 0.655 mmol/L,its sensitivity and specificity for differentiating KD with cardiovascular damage was 80.4％ and 69.4％ respectively.When the blood NT-proBNP was higher than 786.5 ng/L in addition to HDL-C lower than 0.655 mmol/L,the specificity for differentiating KD with cardiovascular damage was 91.8％.③ For the 222 cases with cardiovascular dam-age,their blood NT-proBNP,HDL-C levels were statistically significantly different between the echocardiogra-phy abnormal group and echocardiography normal group (t=3.354,4.084,All P＜0.05).④ The serum NT-proBNP,ALT levels of the 406 acute and convalescent KD patients were significantly higher than the recovery phase.The blood HDL-C,albumin level of acute patients were significantly lower than those at the recovery phase,the difference was statistically significant (t=22.335,4.951,20.334,15.073,All P＜0.05).⑤ One hundred and four children with IKD were divided into patients with cardiovascular damage (74 cases) and without cardiovascu-lar damage (30 cases),the age,lip and oral changes,the first application of intravenous gamma globulin,blood NT-proBNP,HDL-C,albumin and ALT were significantly differentbetween these two groups (t=3.083,2.157,6.423,6.409,3.649,8.658,All P＜0.05).Conclusion Blood NT-proBNP and HDL-C are good pre-dictive parameters in children with cardiovascular damage of KD,IKD.