Objective Currently, there is no commercially available quantitative detection kit for the main Felis domestic allergen (Fel d 1) in China. To establish a rapid detection method for Fel d 1, this study aims to prepare monoclonal antibodies against Fel d 1 protein. Methods The codon preference of Escherichia coli was utilized to optimize and synthesize the Fel d 1 gene. The prokaryotic expression plasmid pET-28a-Fel d 1 was constructed and used to express and purify the recombinant Fel d 1 protein. Subsequently, the recombinant protein was immunized into BALB/c mice and monoclonal antibodies (mAbs) were prepared by the hybridoma technique. An indirect ELISA was established using the recombinant Fel d 1 as the coating antigen, and hybridoma cell lines were screened for positive clones. The specificity and antigenic epitopes of the mAbs were confirmed by Western blot analysis. Finally, the selected hybridoma cells were injected into the peritoneal cavities of BALB/c mice for large-scale monoclonal antibody production. Results The recombinant plasmid pET-28a-Fel d 1 was successfully constructed, and soluble Fel d 1 protein was obtained after optimizing the expression conditions. Western blot and antibody titer assays confirmed the successful isolation of two hybridoma cell lines, 7D11 and 5H4, which stably secreted mAbs specific to Fel d 1. Antibody characterization revealed that the 5H4 mAb was of the IgG2a subtype and could recognize the amino acid region 105-163 of Fel d 1, while the 7D11 mAb was the IgG1 subtype and could recognize the amino acid region 1-59. Conclusion The high-purity recombinant Fel d 1 protein produced in this study provides a promising alternative for clinical immunotherapy of cat allergies. Furthermore, the monoclonal antibody prepared in this experiment lays a material foundation for the in-depth study of the biological function of Fel d 1 and the development of ELISA detection.
Animals ; Antibodies, Monoclonal/biosynthesis* ; Mice, Inbred BALB C ; Cats ; Mice ; Allergens/genetics* ; Glycoproteins/genetics* ; Enzyme-Linked Immunosorbent Assay ; Hybridomas/immunology* ; Recombinant Proteins/genetics* ; Female ; Antibody Specificity

Objective·To investigate the changes in transcriptome and chromatin accessibility during the differentiation of human embryonic stem cells(hESCs)into neural progenitor cells(NPCs)using in vitro differentiation models and high-throughput multi-omics sequencing technologies.Methods·hESCs were first induced to differentiate into NPCs in vitro using the embryoid body formation method,and cells at both stages were collected.The cell phenotypes were identified by reverse transcription-quantitative real-time PCR(RT-qPCR)and immunofluorescence(IF)staining.Transcriptome sequencing(RNA-seq)was conducted to detect and analyze the differentially expressed genes(DEGs)between hESCs and NPCs.The assay for transposase-accessible chromatin with high-throughput sequencing(ATAC-seq)was employed to assess chromatin accessibility changes between hESCs and NPCs.Motif enrichment analysis was performed on differentially accessible chromatin regions to discover potential regulatory transcription factors.Finally,an integrated analysis of RNA-seq and ATAC-seq data and the protein-protein interaction(PPI)network were performed to identify key genes and regulatory pathways involved in the early stages of neural differentiation in vitro.Results·Both RT-qPCR and IF results indicated that the expression levels of pluripotency markers(NANOG and POU5F1)were high at the hESC stage but significantly decreased at the NPC stage,while early neural differentiation markers(PAX6,SOX1,and NES)were minimally expressed at the hESC stage but markedly upregulated at the NPC stage.RNA-seq analysis revealed that compared to the hESC stage,there were 5 597 genes upregulated and 3 654 genes downregulated at the NPC stage.Gene function enrichment analysis showed that the upregulated genes at the NPC stage were enriched in the functions related to neural development.ATAC-seq analysis demonstrated a total of 27 491 genomic regions had significant changes in chromatin accessibility during the differentiation from hESC to NPC,with 12 381 regions showing increased accessibility and 15 110 regions showing decreased accessibility.Motif enrichment analysis revealed that transcription factor genes such as DLX1 and LHX2 might play an important role in the differentiation process from hESCs into NPCs.Integrated analysis of RNA-seq and ATAC-seq data revealed that overlapping genes with high expression at the NPC stage were mainly enriched in axon guidance,forebrain development,and neuron migration.After neural differentiation,the expression levels of CTNND2 and LHX2 genes increased,and the chromatin accessibility of related genomic regions also increased.PPI network analysis indentified candidate downstream genes including PRKACA,CDH2,and ERBB4.Conclusion·The in vitro differentiation model of hESCs combined with high-throughput multi-omics sequencing technologies can be used to depict the changes in transcriptome and chromatin accessibility during the differentiation of hESCs into NPCs.In this process,the expression levels of genes related to axon guidance,forebrain development,and neuronal migration pathways increase and related chromatin accessibility is enhanced.

Objective:To explore the diagnostic value of serum solute carrier family 7 member 11 (SLC7A11), urinary retinol-binding protein (RBP) and transferrin (TRF) for acute kidney injury (AKI) in patients with sepsis.Methods:The clinical data of 204 patients with sepsis from January 2020 to December 2023 in the Second Affiliated Hospital of Xi′an Medical College were retrospectively analyzed. Among them, 102 patients complicated with AKI (AKI group), including Kidney Disease: Improving Global Outcomes (KDIGO) classification Ⅰ stage 43 cases, Ⅱ stage 36 cases, Ⅲ stage 23 cases; 102 patients did not complicate with AKI (non-AKI group). Additionally, 102 healthy individuals from the same period were selected as a healthy control group. Enzyme-linked immunosorbent assay was used to detect the serum expression level of SLC7A11, and fully automatic biochemical analyzers were used to detect urinary RBP and TRF levels. For patients in AKI group and non-AKI group, the sequential organ failure assessment (SOFA) was recorded; fully automatic analyzers were used to test hematological indicators, including creatinine, hemoglobin, platelet, albumin, uric acid, lactate, procalcitonin and C-reactive protein, and estimated glomerular filtration rate (eGFR) was calculated. Multivariate Logistic regression analysis was used to analyze the independent risk factors of AKI in patients with sepsis. Receiver operating characteristic (ROC) curve was plotted to analyze the values of serum SLC7A11 and urinary RBP, TRF in assessing the risk of AKI in patients with sepsis.Results:The serum SLC7A11 and urinary RBP, TRF in non-AKI group and AKI group were significantly higher than those in healthy control group: (28.66 ± 6.22) and (36.18 ± 7.29) ng/L vs. (14.32 ± 2.63) ng/L, (1.20 ± 0.25) and (1.47 ± 0.31) mg/L vs. (0.44 ± 0.08) mg/L, (1.82 ± 0.39) and (2.26 ± 0.45) mg/L vs. (1.08 ± 0.19) mg/L, furthermore the indexes in AKI group were significantly higher than those in non-AKI group, and there were statistical differences ( P<0.05). The serum SLC7A11 and urinary RBP, TRF in patients with KDIGO Ⅱ stage and Ⅲ stage were significantly higher than those in patients with KDIGO Ⅰ stage: (37.16 ± 7.41) and (45.20 ± 8.29) ng/L vs. (30.53 ± 6.46) ng/L, (1.50 ± 0.28) and (1.72 ± 0.35) mg/L vs. (1.31 ± 0.26) mg/L, (2.26 ± 0.46) and (2.77 ± 0.59) mg/L vs. (1.99 ± 0.40) mg/L, furthermore the indexes in patients with KDIGO Ⅲ stage were significantly higher than those in patients with KDIGO Ⅱ stage, and there were statistical differences ( P<0.05). The SOFA, creatinine and lactate in AKI group were significantly higher than those in non-AKI group: 12 (9, 15) scores vs. 7 (5, 9) scores, (133.71 ± 13.58) μmol/L vs. (108.18 ± 14.32) μmol/L and (13.61 ± 3.57) mmol/L vs. (10.95 ± 3.10) mmol/L, the albumin and eGFR were significantly lower than those in non-AKI group: (21.48 ± 2.48) g/L vs. (24.85 ± 2.83) g/L and (51.57 ± 9.64) ml/(min·1.73 m 2) vs. (59.21 ± 10.67) ml/(min·1.73 m 2), and there were statistical differences ( P<0.01); there were no statistical differences in hemoglobin, platelet, uric acid, procalcitonin and C-reactive protein between two groups ( P>0.05). Multivariate Logistic regression analysis result showed that the high SOFA, creatinine, lactate, SLC7A11, urinary RBP, urinary TRF, and low eGFR, albumin were independent risk factors for AKI in patients with sepsis ( OR = 4.864, 5.631, 2.315, 5.862, 6.852, 6.218, 0.328 and 0.226; 95% CI 1.701 to 13.907, 1.803 to 17.585, 1.350 to 3.969, 2.115 to 16.242, 2.177 to 21.566, 1.900 to 20.353, 0.151 to 0.713 and 0.092 to 0.555; P<0.01). The ROC curve analysis result showed that the area under the curve of the combined assessment of serum SLC7A11 and urinary RBP, TRF for AKI in patients with sepsis was significantly larger than serum SLC7A11 and urinary RBP, TRF alone (0.892 vs. 0.774, 0.765 and 0.755), and there was statistical difference ( Z = 2.97, 3.20 and 3.38; P<0.01). Conclusions:The elevated expression levels of serum SLC7A11 and urinary RBP and TRF in patients with sepsis have a high value for the combined detection and assessment of AKI.

Objective:To explore the diagnostic value of serum solute carrier family 7 member 11 (SLC7A11), urinary retinol-binding protein (RBP) and transferrin (TRF) for acute kidney injury (AKI) in patients with sepsis.Methods:The clinical data of 204 patients with sepsis from January 2020 to December 2023 in the Second Affiliated Hospital of Xi′an Medical College were retrospectively analyzed. Among them, 102 patients complicated with AKI (AKI group), including Kidney Disease: Improving Global Outcomes (KDIGO) classification Ⅰ stage 43 cases, Ⅱ stage 36 cases, Ⅲ stage 23 cases; 102 patients did not complicate with AKI (non-AKI group). Additionally, 102 healthy individuals from the same period were selected as a healthy control group. Enzyme-linked immunosorbent assay was used to detect the serum expression level of SLC7A11, and fully automatic biochemical analyzers were used to detect urinary RBP and TRF levels. For patients in AKI group and non-AKI group, the sequential organ failure assessment (SOFA) was recorded; fully automatic analyzers were used to test hematological indicators, including creatinine, hemoglobin, platelet, albumin, uric acid, lactate, procalcitonin and C-reactive protein, and estimated glomerular filtration rate (eGFR) was calculated. Multivariate Logistic regression analysis was used to analyze the independent risk factors of AKI in patients with sepsis. Receiver operating characteristic (ROC) curve was plotted to analyze the values of serum SLC7A11 and urinary RBP, TRF in assessing the risk of AKI in patients with sepsis.Results:The serum SLC7A11 and urinary RBP, TRF in non-AKI group and AKI group were significantly higher than those in healthy control group: (28.66 ± 6.22) and (36.18 ± 7.29) ng/L vs. (14.32 ± 2.63) ng/L, (1.20 ± 0.25) and (1.47 ± 0.31) mg/L vs. (0.44 ± 0.08) mg/L, (1.82 ± 0.39) and (2.26 ± 0.45) mg/L vs. (1.08 ± 0.19) mg/L, furthermore the indexes in AKI group were significantly higher than those in non-AKI group, and there were statistical differences ( P<0.05). The serum SLC7A11 and urinary RBP, TRF in patients with KDIGO Ⅱ stage and Ⅲ stage were significantly higher than those in patients with KDIGO Ⅰ stage: (37.16 ± 7.41) and (45.20 ± 8.29) ng/L vs. (30.53 ± 6.46) ng/L, (1.50 ± 0.28) and (1.72 ± 0.35) mg/L vs. (1.31 ± 0.26) mg/L, (2.26 ± 0.46) and (2.77 ± 0.59) mg/L vs. (1.99 ± 0.40) mg/L, furthermore the indexes in patients with KDIGO Ⅲ stage were significantly higher than those in patients with KDIGO Ⅱ stage, and there were statistical differences ( P<0.05). The SOFA, creatinine and lactate in AKI group were significantly higher than those in non-AKI group: 12 (9, 15) scores vs. 7 (5, 9) scores, (133.71 ± 13.58) μmol/L vs. (108.18 ± 14.32) μmol/L and (13.61 ± 3.57) mmol/L vs. (10.95 ± 3.10) mmol/L, the albumin and eGFR were significantly lower than those in non-AKI group: (21.48 ± 2.48) g/L vs. (24.85 ± 2.83) g/L and (51.57 ± 9.64) ml/(min·1.73 m 2) vs. (59.21 ± 10.67) ml/(min·1.73 m 2), and there were statistical differences ( P<0.01); there were no statistical differences in hemoglobin, platelet, uric acid, procalcitonin and C-reactive protein between two groups ( P>0.05). Multivariate Logistic regression analysis result showed that the high SOFA, creatinine, lactate, SLC7A11, urinary RBP, urinary TRF, and low eGFR, albumin were independent risk factors for AKI in patients with sepsis ( OR = 4.864, 5.631, 2.315, 5.862, 6.852, 6.218, 0.328 and 0.226; 95% CI 1.701 to 13.907, 1.803 to 17.585, 1.350 to 3.969, 2.115 to 16.242, 2.177 to 21.566, 1.900 to 20.353, 0.151 to 0.713 and 0.092 to 0.555; P<0.01). The ROC curve analysis result showed that the area under the curve of the combined assessment of serum SLC7A11 and urinary RBP, TRF for AKI in patients with sepsis was significantly larger than serum SLC7A11 and urinary RBP, TRF alone (0.892 vs. 0.774, 0.765 and 0.755), and there was statistical difference ( Z = 2.97, 3.20 and 3.38; P<0.01). Conclusions:The elevated expression levels of serum SLC7A11 and urinary RBP and TRF in patients with sepsis have a high value for the combined detection and assessment of AKI.

OBJECTIVE To investigate the serotypes and antimicrobial resistance rate of clinically isolated Salmonel-la in a children's hospital in Suzhou,and to provide reference for the treatment of salmonellosis.METHOD Totally 177 strains of Salmonella isolated from Children's Hospital of Wujiang District from Jan.2021 to Dec.2023 were collected,and the results of serotypes and drug sensitivity of Salmonella were analyzed.RESULTS The male to fe-male isolation rate of Salmonella was 1.39∶1,with a median age of children infection at 1.3(0.8,2.3)years.The highest number of Salmonella strains were isolated in the month of Jun.,followed by Jul.,Aug.,Sep.,Oct.and May,collectively accounting for 82.49％of all isolates.Acute gastroenteritis was manifested in 142 cases(80.22％),with respiratory tract infections in 38 cases(21.47％)and septicemia in 7 cases(3.95％).The differ-ence in detection rates across the three years was not statistically significant(P=0.806).Salmonella Typhimuri-um was the predominant serotype,representing 54.24％of all isolates.The antimicrobial drug with the highest rate of resistance in Salmonella was ampicillin(71.35％,122/171),followed by sulfamethoxazole/metronidazole(43.60％,75/172),and ampicillin/sulbactam(30.23％,52/172),and no imipenem resistant strains were found.29.07％(50/172)of strains showed multidrug resistance.CONCLUSIONS Boys under three years of age are sus-ceptible to Salmonella infections in summer and fall in this region,with Salmonella Typhimurium being the pre-dominant serotype.Clinical attention should be paid to the characteristics of Salmonella infection and drug resist-ance,as well as the early diagnosis and rational use of antimicrobial drugs.

Objective·To investigate the changes in transcriptome and chromatin accessibility during the differentiation of human embryonic stem cells(hESCs)into neural progenitor cells(NPCs)using in vitro differentiation models and high-throughput multi-omics sequencing technologies.Methods·hESCs were first induced to differentiate into NPCs in vitro using the embryoid body formation method,and cells at both stages were collected.The cell phenotypes were identified by reverse transcription-quantitative real-time PCR(RT-qPCR)and immunofluorescence(IF)staining.Transcriptome sequencing(RNA-seq)was conducted to detect and analyze the differentially expressed genes(DEGs)between hESCs and NPCs.The assay for transposase-accessible chromatin with high-throughput sequencing(ATAC-seq)was employed to assess chromatin accessibility changes between hESCs and NPCs.Motif enrichment analysis was performed on differentially accessible chromatin regions to discover potential regulatory transcription factors.Finally,an integrated analysis of RNA-seq and ATAC-seq data and the protein-protein interaction(PPI)network were performed to identify key genes and regulatory pathways involved in the early stages of neural differentiation in vitro.Results·Both RT-qPCR and IF results indicated that the expression levels of pluripotency markers(NANOG and POU5F1)were high at the hESC stage but significantly decreased at the NPC stage,while early neural differentiation markers(PAX6,SOX1,and NES)were minimally expressed at the hESC stage but markedly upregulated at the NPC stage.RNA-seq analysis revealed that compared to the hESC stage,there were 5 597 genes upregulated and 3 654 genes downregulated at the NPC stage.Gene function enrichment analysis showed that the upregulated genes at the NPC stage were enriched in the functions related to neural development.ATAC-seq analysis demonstrated a total of 27 491 genomic regions had significant changes in chromatin accessibility during the differentiation from hESC to NPC,with 12 381 regions showing increased accessibility and 15 110 regions showing decreased accessibility.Motif enrichment analysis revealed that transcription factor genes such as DLX1 and LHX2 might play an important role in the differentiation process from hESCs into NPCs.Integrated analysis of RNA-seq and ATAC-seq data revealed that overlapping genes with high expression at the NPC stage were mainly enriched in axon guidance,forebrain development,and neuron migration.After neural differentiation,the expression levels of CTNND2 and LHX2 genes increased,and the chromatin accessibility of related genomic regions also increased.PPI network analysis indentified candidate downstream genes including PRKACA,CDH2,and ERBB4.Conclusion·The in vitro differentiation model of hESCs combined with high-throughput multi-omics sequencing technologies can be used to depict the changes in transcriptome and chromatin accessibility during the differentiation of hESCs into NPCs.In this process,the expression levels of genes related to axon guidance,forebrain development,and neuronal migration pathways increase and related chromatin accessibility is enhanced.

OBJECTIVE To investigate the serotypes and antimicrobial resistance rate of clinically isolated Salmonel-la in a children's hospital in Suzhou,and to provide reference for the treatment of salmonellosis.METHOD Totally 177 strains of Salmonella isolated from Children's Hospital of Wujiang District from Jan.2021 to Dec.2023 were collected,and the results of serotypes and drug sensitivity of Salmonella were analyzed.RESULTS The male to fe-male isolation rate of Salmonella was 1.39∶1,with a median age of children infection at 1.3(0.8,2.3)years.The highest number of Salmonella strains were isolated in the month of Jun.,followed by Jul.,Aug.,Sep.,Oct.and May,collectively accounting for 82.49％of all isolates.Acute gastroenteritis was manifested in 142 cases(80.22％),with respiratory tract infections in 38 cases(21.47％)and septicemia in 7 cases(3.95％).The differ-ence in detection rates across the three years was not statistically significant(P=0.806).Salmonella Typhimuri-um was the predominant serotype,representing 54.24％of all isolates.The antimicrobial drug with the highest rate of resistance in Salmonella was ampicillin(71.35％,122/171),followed by sulfamethoxazole/metronidazole(43.60％,75/172),and ampicillin/sulbactam(30.23％,52/172),and no imipenem resistant strains were found.29.07％(50/172)of strains showed multidrug resistance.CONCLUSIONS Boys under three years of age are sus-ceptible to Salmonella infections in summer and fall in this region,with Salmonella Typhimurium being the pre-dominant serotype.Clinical attention should be paid to the characteristics of Salmonella infection and drug resist-ance,as well as the early diagnosis and rational use of antimicrobial drugs.

In recent years, with the rapid development of organ donation after citizen’s death and transplantation, central and local governments in China have successively released incentive policies. To protect the legitimate rights and interests of organ donors after citizen’s death and their families, current status of incentive policies for organ donation after citizen’s death was illustrated and analyzed from the perspective of ethics. Combining with the principles of justice, respect for autonomy, nonmaleficence and beneficence, the problems existing in the implementation of incentive policies for organ donation after citizen’s death were identified in China, such as lack of continuous psychological intervention in spiritual incentives, the misinterpretation of humanitarian assistance in practice and the impact of indirect economic incentives on individual donation autonomy, etc. Relevant countermeasures and suggestions were proposed at the government, society and individual levels, aiming to provide reference for improving the incentive policies for organ donation after citizen’s death and accelerate the development of organ donation in China.

Objective:To comprehensively analyze the clinical characteristics,classification of causes,treatment methods,and outcomes of pediatric convulsions in the emergency department,providing a scientific basis for the diagnosis and treatment of pediatric convulsions and optimizing emergency management for these cases.Methods:The clinical data of 18 217 children with convulsions in the emergency department of Capital Institute of Pediatrics' Children's Hospital from January 1,2016 to December 31,2020 were retrospectively analyzed.Results:A total of 18 217 children were admitted to the emergency department due to convulsions,accounting for 2.3% of all visits.Among them,58.7% cases were male and 41.3% cases were female,with an average age of (2.00±0.03) years.The main age was 1 to 3 years old (54.2%).Generalized convulsions were the primary type (82.3%),with focal seizures accounting for 17.7%.Most convulsions lasted less than 5 minutes (82.4%),and approximately 55.2% of the patients could self-resolve.Febrile convulsions were the primary cause (69.2%),followed by benign convulsions with mild gastroenteritis (11.7%) and epilepsy (10.5%).Regarding treatment,54.0% of the children recovered without medication.In the triage system of "three zones and four levels," the usage rate of anticonvulsants in the red zone was 93.1%,with 21.6% requiring combined treatment.After treatment,48.2% of the children returned home,92.2% showed improvement or recovery,and the mortality rate was extremely low at only 0.03%.Conclusion:Febrile convulsions are the main cause of pediatric convulsions in the emergency department,and an efficient triage system play an important role in improving treatment response.Different treatment zones and outcomes vary,providing important reference for optimizing emergency management.

ObjectiveTo establish a non-targeted screening method for emerging contaminants in drinking water based on high-resolution mass spectrometry and apply it to actual water samples. MethodsA total of 9 drinking water samples collected from 3 reservoirs in Shanghai were purified and concentrated by HLB solid phase extraction column， then separated and analyzed by liquid chromatography high-resolution mass spectrometer and gas chromatography high⁃resolution mass spectrometer. The acquired data were analyzed by Thermo Tracefinder， Excel and other software combined with mzCloud and NIST databases. The methodology was verified with representative compound standards. Pesticide and perfluorinated compounds were taken as examples to analyze their pollution status. ResultsA non-targeted analysis strategy based on liquid chromatography and gas chromatography tandem high-resolution mass spectrometry was established. The pollution level of 20 kinds of pesticides and 4 kinds of perfluorinated compounds identified in 9 drinking water samples were higher in the Huangpu River than in the Yangtze River estuary. ConclusionThe established non-targeted screening method by high-resolution mass spectrometry can detect potential emerging contaminants in drinking water without relying on the standards， which provides a powerful technical means for water quality monitoring and risk assessment.