To develop a method for determining the antibody-dependent cell-mediated phagocytosis (ADCP) activity of human epidermal growth factor receptor 2 (HER2)-targeted antibody drug conjugate (ADC) based on the reporter gene assay, we established an ADCP activity assay with Jurkat/NFAT/FcγRIIa cells as the effector cells and BT474 as the target cells. Then, the target cell density, the ratio of effector to target cells, the target cell adhesion time, the incubation time for drug administration, and the induction time after adding effector cells were optimized by the method of design of experiment (DOE). The method showed a significant dose-response relationship, which was complied with the four-parameter equation: y=(A-D)/[1+(x/C)^B]+D. The durability ranges of the target cell density, the ratio of effector to target cells, the target cell adhesion time, the incubation time for drug administration, and the induction time after adding effector cells were (2.5-4.0)×10⁵ cells/mL, 3-5, 1.0-2.0 h, 0 h, and 5.0-6.0 h, respectively. The results of the methodological validation showed that the linear equation was y=1.106 8x-0.011 6, r=0.969 2. The established method showed the relative accuracy ranging from -6.59% to 2.98% and the geometric coefficient of variation less than 11% in the intermediate precision test. Furthermore, the method was target-specific. The method was then applied to the determination of ADCP activity of HER2-targeted ADC, demonstrating the result of (103.5±5.7)%. We developed a reporter gene assay for determining the ADCP activity of HER2-targeted ADC and the assay demonstrated high accuracy and good reproducibility, which proposes a highly efficient and approache for evaluating ADCP effect of this HER2-targeted ADC, and also provides a referable technique for characterizing the Fc effector functions of ADCs with diverse targets.
Humans ; Receptor, ErbB-2/immunology* ; Phagocytosis/drug effects* ; Immunoconjugates/immunology* ; Genes, Reporter ; Antibody-Dependent Cell Cytotoxicity ; Jurkat Cells

Malignant pleural effusion(MPE)is a common complication in patients with advanced malignant tumors,which not only significantly reduces their quality of life but also shortens their survival duration.Despite the widespread use of traditional treatment methods such as thoracentesis and pleurodesis,their efficacy is limited accompanied by high recurrence rates.Therefore,exploring novel therapeutic strategies becomes particularly urgent.In recent years,immunotherapy has attracted extensive attention for its potential in cancer treatment.This article systematically reviews the roles of CD4+T cell subsets,including regulatory T cells(Treg),T helper cell(Th)17,Th9,and Th22 cells,within the immunosuppressive microenvironment of MPE.These cell subsets are involved in the formation of the immunosuppressive state of MPE through various mechanisms and play key roles in the occurrence and development of the disease.In addition,the article discusses in detail the role of immune checkpoint molecules,such as programmed death protein 1(PD-1),PD-1 ligand(PD-L1),and cytotoxic T-lymphocyte-associated protein 4(CTLA-4),in the immune evasion of MPE.The abnormal expressions of these molecules provide opportunity for tumor cells to evade immune system surveillance.At the same time,this article also summarizes the application prospects of novel immunotherapy strategies,such as adoptive cell therapy(ACT)and chimeric antigen receptor T cell(CAR-T)therapy,in the treatment of MPE.These innovative therapies offer possibilities for improving the prognosis of MPE patients through activating and enhancing the anti-tumor immune response.

Objective To investigate the association of neuregulin 4(NRG4)and meteorin-like protein(Metrnl)with insulin resistance in patients with metabolic syndrome(MS).Methods From September to November 2023,totally 60 MS patients in Anhui No.2 Provincial People's Hospital were selected as MS group,and 60 physical examination healthy people were selected as control group.The changes of human mor-phological indicators,biochemical indicators,blood routine,NRG4,Metrnl,insulin resistance index(HOMA-IR)and other indicators in the two groups were observed,and the correlation between NRG4,Metrnl and HO-MA-IR and each index was analyzed,and the diagnostic value of NRG4 and Metrnl for MS was evaluated.Re-sults Compared with the control group,the levels of white blood cell count(WBC),alanine aminotransferase(ALT),uric acid(UA),triglyceride(TG),total cholesterol(TC),fasting insulin(FINS),HOMA-IR,body weight,body mass index(BMI),waist circumference,hip circumference,waist-hip ratio(WHR),and body fat ratio were significantly increased in MS group(P<0.05),however,high density lipoprotein cholesterol(HDL-C),NRG4,and Metrnl were significantly decreased(P<0.05).Spearman correlation analysis showed that serum NRG4 was positively correlated with Metrnl and HDL-C(P<0.05),and negatively correlated with TG,FINS,HOMA-IR,body weight,waist circumference,hip circumference,WHR,and ALT(P<0.05).Serum Metrnl was positively correlated with NRG4 and HDL-C(P<0.05),and negatively correlated with TG and hip circumference(P<0.05).Binary Logistic regression analysis showed that NRG4 and Metrnl were protective factors for MS(P<0.05).The receiver operating characteristic curve analysis showed that the sensitivity of NRG4,Metrnl,and their combination for diagnosing MS was 67%,41%,and 67%,respec-tively,the specificity was 71%,95%,and 86%,respectively,and the area under the curve was 0.713,0.635,and 0.787,respectively.Conclusion The levels of serum NRG4 and Metrnl are decreased in MS patients,and NRG4 is associated with insulin resistance.The combination of NRG4 and Metrnl has a certain diagnostic val-ue for MS.

Objective To investigate the clinical efficacy and safety of endoscopic ultrasonography-guided(EUS-guided)enterocolon anastomosis in treating patients with malignant bowel obstruction(MBO).Methods The clinical data of 12 patients with MBO,who underwent EUS-guided enterocolon anastomosis at the Nanjing Drum Tower Hospital of China from April 2023 to December 2023,were collected.The perioperative clinical efficacy and safety were retrospectively analyzed.Results Successful EUS-guided enterocolon anastomosis was accomplished in all the 12 patients,with a technical success rate of 100％(12/12).The clinical success rate was 83.3％(11/12),one patient developed obstruction of the stent.The clinical symptoms were relieved in 2-68 hours after treatment,and the time to resume defecation and exhaust was(18.02±15.75)hours.Within one week after the operation,4 patients took liquid diet and 8 patients took semi-fluid diet.Each dimension score of the Quality of Life Core-30 scale of The European Organization for Research and Treatment of Cancer(EORTC QLQ-C30)was remarkably improved,the patient's overall health score was increased from preoperative median 5 points to postoperative 8 points(P＜0.001).During the operation,stent displacement occurred in 2 patients,and the operation was successfully completed after promptly taking remedial measures.After operation,11 patients developed fever(37.5-39.4 ℃),and all the patients were discharged smoothly after symptomatic treatment.No complication such as bleeding,perforation,or stent displacement occurred.Conclusion EUS-guided enterocolon anastomosis is clinically safe and effective,it can effectively relieve the clinical symptoms and improve the quality of life of patients with MBO.

Objective:To investigate the current application of blood purification in the treatment of acute poisoning within Jiangsu Province and to evaluate the impact of extracorporeal blood purification on the clinical outcomes of critically poisoned patients.Methods:This multicenter, cross-sectional real-world observational study followed patients presenting with poisoning to the emergency departments of nine hospitals in Jiangsu Province between June 2015 and May 2019. Data were collected on demographic characteristics, vital signs within the first hour of emergency presentation, treatment modalities, length of hospital stay, and survival outcomes. Clinical data from patients who underwent extracorporeal blood purification were compared with those who did not, using the Wilcoxon rank-sum test and Chi-square test.Results:A total of 4 178 poisoning cases were included between June 2015 and May 2019. Among them, 21.7% (908/4 178) received blood purification therapy, while 78.3% (3 270/4 178) did not. Hemoperfusion (90.4%) was the most frequently employed method, followed by continuous renal replacement therapy (CRRT) (4.4%). In combined blood purification modalities, 4.8% underwent hemoperfusion combined with CRRT, 0.1% received hemoperfusion with plasma exchange, and another 0.1% underwent hemoperfusion combined with both CRRT and plasma exchange. Among patients who underwent blood purification, pesticide poisoning was the most prevalent (76.3%), with the most common toxic agents being paraquat (23.7%), dichlorvos (8.7%), methamidophos (5.2%), omethoate (4.0%), and glyphosate (3.7%). Compared to the non-blood purification group, patients in the blood purification group were more likely to present within the first hour with a low Glasgow Coma Scale (GCS) score (3-8) (22.6% vs. 9.7%, P <0.05), low mean arterial pressure (8.0% vs. 3.2%, P <0.05), longer hospital stays [5(3,9) days vs. 2(1,4) days, P <0.05] and a higher in-hospital mortality rate (21.1% vs. 5.3%, P <0.05). Follow-up via telephone 28 days after discharge revealed a survival rate of 78.9%, with a mortality rate of 21.1% in the blood purification group. Conclusions:Hemoperfusion is the most commonly utilized blood purification technique for treating poisoning in Jiangsu Province, with pesticides being the primary toxic agents treated. Although the mortality rate is higher in the blood purification group, the intervention may still contribute to improved patient outcomes.

Objective To investigate the efficacy of laparoscopic C-type radical hysterectomy through deep uterine vein approach for the treatment of cervical cancer.Methods From January 2021 to December 2022,58 cases of cervical cancer were treated with deep uterine vein approach laparoscopic C-type radical hysterectomy in our hospital.After establishing the operation channel,the ureter was identified in the posterior lobe of the broad ligament,and the uterine artery was exposed after separating the ureter.The connective tissue was separated along the dorsal side of the uterine artery to gradually expose the deep uterine vein.The parametrial lymph nodes surrounding the deep uterine vein were resected and sent to pathology alone.The deep uterine vein was continuously tracked towards the bladder,and its branches were freed.The deep uterine vein and its tributaries were double clipped by vascular clamp.Results The operation time was(307.2±54.1)min,the median bleeding volume was 50(20,100)ml,the lymph node dissection number was(26.3±6.9),the indwelling catheter time was(20.6±4.7)d,the time to removal of abdominal drainage tube was(9.4±4.1)d,the anal exhaust time was(36.7±4.1)h,the antibiotics use time was(9.2±4.2)d,and the hospital stay was(13.4±2.6)d.The postoperative complication rate was 3.4%(2/58),and the postoperative pathological staging upgrade rate was 22.4%(13/58).The postoperative European Organization of Research and Treatment of Cancer(EORTC)Quality of Life Questionnaire-Core 30(QLQ-C30)score was significantly higher than before surgery[(78.6±10.7)points vs.(47.1±7.6)points,t = 17.177,P = 0.000].All the 58 cases were followed up for 4-25 months(mean,13.5±6.2 months),with no recurrence.Conclusion Laparoscopic C-type radical hysterectomy through deep uterine vein approach is effective,safe,and reliable.

Objective:To investigate the safety and effectiveness of endoscopic retrograde cholangiopancreatography (ERCP) for the diagnosis and treatment of pediatric pancreaticobiliary maljunction (PBM).Methods:Data of 40 pediatric patients under 14 with PBM diagnosed and treated by ERCP at Department of Gastroenterology, Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School from November 2012 to September 2022 were collected. PBM types, ERCP-related diagnosis and treatment, adverse events and prognosis were retrospectively analyzed.Results:Nineteen cases were P-B type (joining of common bile duct with pancreatic duct), 17 were B-P type (joining of pancreatic duct with common bile duct), and 4 were complex type. Forty children with PBM underwent 50 ERCP-related operations, among which 48 procedures succeeded. One case failed during cannulation of ERCP, replaced by rendezvous-assisted endoscopic retrograde pancreatography (RV-ERP) afterwards. There were no serious postoperative adverse events such as bleeding, perforation or death. Thirty-four patients (85%) were followed up successfully, among which 14 underwent further surgery and 20 continued conservative treatment.Conclusion:ERCP is the golden standard to diagnose pediatric PBM, and it is also safe and effective treatment for PBM.

BACKGROUND:Mesenchymal stem cells are susceptible to senescence during in vitro expansion,which greatly hinders their application in vivo and in vitro.How to improve the replicative senescence of mesenchymal stem cells is an urgent problem to be solved in tissue engineering. OBJECTIVE:To determine whether vascular endothelial growth factor combined with basic fibroblast growth factor can improve the aging of bone marrow mesenchymal stem cells caused by replicative passage. METHODS:Rat bone marrow mesenchymal stem cells were extracted by whole bone marrow adhesion method.Passage 2 cells were selected as normal control group.Passage 7 and later algebraic cells were selected as aging model group.Vascular endothelial growth factor(50 μg/L),basic fibroblast growth factor(10 μg/L),and their combination were administered.Cell proliferation was detected by CCK-8 assay.Cell senescence was observed by β-galactosidase activity staining.Cytoskeleton size and colony formation ability were observed by phalloidine staining and Giemsa staining,respectively,and the levels of senescence-related genes P16,P21,and P53 were detected by qRT-PCR.Gene expression levels of P16,P21,and P53 were tested by qRT-PCR. RESULTS AND CONCLUSION:(1)Vascular endothelial growth factor combined with basic fibroblast growth factor could promote the proliferation of aged bone marrow mesenchymal stem cells,which began to enter the plateau stage on day 9,and the absorbance value of the combined intervention group was significantly higher than that of the model group on day 9(P<0.05).(2)The phenotypic markers of the cells in the combined intervention group did not change,and the cell morphology changed from broad to slender.(3)Compared with the model group,the positive rate of β-galactosidase was significantly decreased(P<0.01);the number of nuclei increased(P<0.001);the total area of cytoskeleton increased(P<0.01);colony formation ability was enhanced(P<0.05);expression level of P16 was decreased(P<0.01)in the combined intervention group.These results indicate that vascular endothelial growth factor combined with basic fibroblast growth factor can improve the senescence of bone marrow mesenchymal stem cells caused by replicative passage without changing the cell phenotype.

Objective:To explore the clinical phenotype and pathogenesis of a child with partial duplication in the long arm of chromosome 10 (10q), and conduct a review of relevant literature.Methods:A child presented at Lianyungang Maternal and Child Health Care Hospital in April 2018 for growth retardation, intellectual disability, and autism spectrum disorder (ASD) was selected as the study subject. Peripheral blood samples were collected from the child and his parents for G-banded chromosomal karyotyping analysis. Genomic DNA was also extracted for chromosomal microarray analysis (CMA). The clinical phenotype and relevant genes were searched in the Online Mendelian Inheritance in Man (OMIM) and the UK Database of Genomic Variation and Phenotype in Humans using Ensembl Resources (DECIPHER). The pathogenicity of chromosomal variation was analyzed based on guidelines from the American College of Medical Genetics and Genomics (ACMG). Relevant literature was searched from the CNKI, Wanfang Data, and PubMed databases by using keywords such as " 10q" " duplication" and " trisomy", with the time set as from the establishment of database to December 1, 2023. This study has been approved by the Medical Ethics Committee of the Lianyungang Maternal and Child Health Care Hospital (No. XM2023030).Results:The clinical phenotype of child had included growth retardation, intellectual disability, and ASD. G-banded chromosomal analysis suggested that the child has a karyotype of 46, XY, dup(10)(q23.31q24.33), whilst both of his parents were normal. CMA analysis of the child revealed that the child was arr[19]10q23.31q24.33(87603382_104948862)×3, with a 17.34 Mb duplication in the 10q23.31q24.33 region. Search of the OMIM database suggested that the duplicated segment has contained 171 genes associated with various diseases, and search of the DECIPHER database has identified cases with overlapping with the duplication. A search of the PubMed database has identified 2 publications involving 2 patients with chromosomal duplications overlapping the 10q23.31q24.33 region with a segment length of > 10 Mb. The 2 patients had mainly manifested growth retardation, intellectual disability, ASD, and facial and limb malformations. The main pathogenic genes had included PTEN, WNT8B, LZTS2, NFKB2, PAX2, KIF11, FRA10AC1, and CNNM2. No similar case was retrieved from the CNKI and Wanfang Data databases. Conclusion:The partial 10q duplication as a novel CNV involving genes such as PTEN and WNT8B probably underlay the growth retardation, intellectual disability and ASD in the child. This study has enriched the genotype-phenotype spectrum of patients with partial 10q23.31q24.33 duplications.

Objective:To explore the correlation between multidimensional nutritional evaluation indexes of folate and vitamin B 12 and mild cognitive impairment in community-dwelling older adults. Methods:In this study, cross-sectional study design was used, and the elderly people aged 60 years and above were recruited from three parallel communities in Shijiazhuang City.The survey was conducted including basic information questionnaire and semi-quantified food frequency questionnaires(FFQ)in the past year.The Montreal Cognitive Assessment Scale(MoCA)assessed the cognitive function.The subjects were divided into two groups of normal cognition and mild cognitive impairment according to the criteria categorized by education levels recommended by the 2018 China Dementia and Cognitive Impairment Diagnosis and Treatment Guidelines.Serum folate, vitamin B 12 and erythrocyte folate levels were measured by electrochemiluminescence immunoassay, serum homocysteine concentrations were measured by circulating enzyme assay, dietary folate and vitamin B 12 intake was calculated by FFQ survey and food composition tables.Logistic regression models were used to analyze the association between multidimensional nutritional indicators of folate and vitamin B 12 and mild cognitive impairment. Results:A total of 537 subjects were included in the study, involving 379(70.6%)with normal cognition and 158(29.4%)with mild cognitive impairment.Dietary vitamin B 12 intake and erythrocyte folate concentration was significantly lower in the elderly with mild cognitive impairment than in those with normal cognition[median(interguartile): 18.9(11.0, 32.9)μg/d vs.20.1(14.2, 34.8)μg/d, Z=2.010, P=0.044; median: 359.4(304.2, 413.4)μg/L vs.378.4(322.5, 433.1)μg/L, Z=2.168, P=0.031].The elderly with mild cognitive impairment had a higher prevalence of folate deficiency than those with normal cognition[17(10.8%) vs.22(5.8%), χ2=4.065, P=0.044].After adjusting for age, sex, and energy intake, the OR(95% CI)values for dietary vitamin B 12 intake and erythrocyte folate concentration in relation to mild cognitive impairment in the elderly were 0.99(0.987, 0.996)( P=0.039)and 0.98(0.974, 0.993)( P=0.021), respectively.The OR(95% CI)values for folate deficiency for mild cognitive impairment was 1.96(1.009, 3.795)( P=0.046). Conclusions:Long-term high dietary vitamin B 12 intake and better in vivo nutritional deposition of folate were negatively associated with mild cognitive impairment in community-dwelling elderly adults, whereas indicators reflecting the short-term nutritional status of folate and vitamin B 12 may be less associated with mild cognitive impairment.