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 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.

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.

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.

Objective·To study the relationship between evolution and the developmental process from the perspective of DNA sequence conservation,and explore their inherent principles.Methods·First,conservation rate(CR)was established by analyzing the conservation of amino acid sequences of coding genes in 100 species to quantify the evolutionary conservation of genes.The relationship between CR and developmental potential was verified by using the feature genes involved in embryonic stem cells pathways.Secondly,cell type-specific genes and their characteristics in conservation were studied by analyzing the RNA sequencing(RNA-seq)data of the three early germ layers(ectoderm,mesoderm and endoderm)and their corresponding mature organs(brain,heart,liver,etc).Then,chromatin immunoprecipitation sequencing(ChIP-seq)data of enhancer histone H3 acetylated at lysine 27(H3K27ac)from early germ layers and mature organs were collected to search for enhancer sites and identify super enhancers in various cells and tissues by using the ROSE procedure.Functional enrichment and signaling pathway analysis of genes was used to examine the identity correlation between SEs-regulated genes and the corresponding cell characteristics,to clarify whether the SEs identified in this study were consistent with the characteristics reported in previous studies.Finally,PhastCons program was used to calculate the DNA conservation score(CS)of non-coding regulatory regions to study their relationship with developmental potential.Results·In the coding region of DNA,CR was successfully established to quantify the conservation of genes.The gene expression data of early germ layers and mature organs showed that the genes with higher conservation rate were more relevant to the stemness and early developmental process,and the differences between the tissues from early and late development could be distinguished by using CR.In the non-coding regions of DNA,it was found that the conservation of regulatory regions was also correlated with development.The CS of the SE sequences in the early developmental germ layers was significantly higher than that of the SE sequences in the corresponding mature organs.However,cell-specific typical enhancers(TEs)did not show such a trend.Conclusion·During the developmental process,CR of genes expressed in the coding region decreases,and CS of super-enhancer DNA in the non-coding region decreases.

Objective:To investigate the clinical characteristics and possible causes of transient spontaneous remission of childhood acute leukemia.Methods:The data of 3 children with acute leukemia who had transient spontaneous remission before standardized chemotherapy in Sun Yat-sen Memorial Hospital of Sun Yat-sen University in July 2018, May 2019 and October 2020 were collected. Moreover, the related influencing factors of spontaneous remission in leukemia were discussed by review of the literature.Results:All 3 children had fever at the onset of the disease, and they achieved transient spontaneous remission after anti-infection therapy. Case 1 obtained partial remission after the initial diagnosis of acute B lymphocytic leukemia (B-ALL), leukemia gene test showed E2A-PBX1 fusion, and relapsed after 12 days. Case 2 obtained spontaneous remission after the initial diagnosis of B-ALL, leukemia gene test showed p16 gene deletion and NRAS and EP300 genes mutation, and relapsed after 20 days. Case 3 obtained spontaneous remission after the initial diagnosis of acute monocytic leukemia, leukemia gene test showed MLL-ENL fusion and NRAS gene mutation, and relapsed after 30 days. A review of the literature showed that the main influencing factors of spontaneous remission in leukemia were Down syndrome, infection and blood transfusion. Other influencing factors included leukemia-related genes, termination of pregnancy and application of drugs.Conclusions:Transient spontaneous remission of childhood acute leukemia is rare in clinical practice, and the possible mechanism is related to infection-induced immune abnormalities. It is recommended that leukemia patients with spontaneous remission should be closely monitored for minimal residual disease.

The importance of structural variants(SVs)for human phenotypes and diseases is now recognized.Although a variety of SV detection platforms and strategies that vary in sensitivity and specificity have been developed,few benchmarking procedures are available to confidently assess their performances in biological and clinical research.To facilitate the validation and application of these SV detection approaches,we established an Asian reference material by characterizing the genome of an Epstein-Barr virus(EBV)-immortalized B lymphocyte line along with identified benchmark regions and high-confidence SV calls.We established a high-confidence SV callset with 8938 SVs by integrating four alignment-based SV callers,including 109x Pacific Biosciences(PacBio)continuous long reads(CLRs),22 x PacBio circular consensus sequencing(CCS)reads,104x Oxford Nanopore Technologies(ONT)long reads,and 114×Bionano optical mapping plat-form,and one de novo assembly-based SV caller using CCS reads.A total of 544 randomly selected SVs were validated by PCR amplification and Sanger sequencing,demonstrating the robustness of our SV calls.Combining trio-binning-based haplotype assemblies,we established an SV benchmark for identifying false negatives and false positives by constructing the continuous high-confidence regions(CHCRs),which covered 1.46 gigabase pairs(Gb)and 6882 SVs supported by at least one diploid haplotype assembly.Establishing high-confidence SV calls for a benchmark sample that has been characterized by multiple technologies provides a valuable resource for investigating SVs in human biology,disease,and clinical research.

Objective:To investigate the role of lncRNA H19 in evaluating prognosis and regulating radioresistance of colon cancer, aiming to provide a new potential target for the diagnosis and treatment of colon cancer.Methods:The value of lncRNA H19 in the clinicopathological parameters and prognosis of colon cancer was assessed based on bioinformatics technology. The expression of lncRNA H19 in HCT116 and SW620 cells was regulated through siRNA and overexpression plasmid transfection, respectively. The effect of regulating lncRNA H19 expression on the proliferation, DNA synthesis, radiosensitivity and cell cycle of colon cancer cells after X-ray irradiation were detected by CCK8, EdU, cell clonogenic survival assay and flow cytometry.Results:The expression of lncRNA H19 was significantly upregulated in colon cancer tissues and correlated with poor prognosis in colon cancer patients. LncRNA H19, as a high-risk gene for colon cancer, had a significant advantage for prognostic assessment of colon cancer (AUC=0.816). Furthermore, the expression of lncRNA H19 was upregulated after X-ray irradiation in colon cancer cells. Knockdown of lncRNA H19 (siRNA-H19) significantly increased the radiosensitivity in HCT116 cells, while overexpression of lncRNA H19 (H19-OE) enhanced the radioresistance in SW620 cells. Moreover, flow cytometry revealed that the G 2/M phase arrest induced by X-ray irradiation was obviously aggravated after siRNA-H19 treatment in colon cancer cells, which suggested that lncRNA H19 might regulate the radiosensitivity by inhibiting cell cycle progression. Conclusion:LncRNA H19 plays a key role in the prognostic assessment and regulating the radiosensitivity in colon cancer, which can be used as a potential target for improving radiosensitivity of colon cancer radiotherapy.

ObjectiveTo study the relationship between the exposure to two kinds of phthalate esters （PAEs） ［Di-N-butyl phthalate，（DBP） and Di-（2-ethylhexyl）phthalate （DEHP）］ and estrogen homeostasis in pregnant women. MethodsIn 2021， we classified the Jiading District of Shanghai into five geographical areas， east， west， south， north and central. A total of 151 pregnant women from each area were selected for questionnaire survey， with random urine samples during first， second， and third trimesters collected. A DBP metabolite ［Mono-N-butyl phthalate （MBP）］ and two DEHP metabolites ［Mono（2-ethylhexyl） phthalate （MEHP）， Mono（2-ethyl5-oxohexyl） phthalate， （MEOHP）］ and three estrogens ［estrone （E₁）， 17β -estradiol （E₂）， and estriol （E₃）］ in urine were determined by ultra-performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry. After a natural logarithmic transformation of PAEs metabolite levels and estrogen concentration， multivariable linear regression was used to control potential confounders and determine the relationship between PAEs metabolite levels and estrogen concentration. ResultsThe detection rates of three PAEs metabolites in urine of pregnant women were more than 98%. The median corrected concentrations of MBP， MEHP and MEOHP were 5.18， 0.59 and 4.23 mg·kg^-1， respectively. During the whole pregnancy， MEOHP was positively correlated with E₁ （β=0.450， 95%CI： 0.057‒0.844）， and MBP was positively correlated with E₃ （β=0.250， 95%CI： 0.034‒0.465）. Stratified by trimesters， MBP was positively correlated with E₃ in the first trimester （β=0.428， 95%CI： 0.103‒0.752）. MEOHP was positively correlated with E₁ in the second trimester （β=0.734， 95%CI： 0.130‒0.752）， and had a possitive trend with E₁ in the third trimester （β=0.744， 95%CI： -0.140‒1.629）. In addition， MEHP had a negative correlation with E₁ in the second trimester （β=-0.498， 95%CI： -1.063‒0.066）. MEOHP had a positive correlation trend with E₂ （β=0.628， 95%CI： -0.101‒1.356） in the third trimester. ConclusionPAEs exposure may interfere with estrogen homeostasis during pregnancy and differs by trimesters. Given the cross-sectional nature of this study， it warrants further study to validate the findings.

Objective:To investigate the correlation between mental health and symptom distress in breast cancer patients.Methods:The clinical data of 110 female breast cancer patients at Shanxi Bethune Hospital from June 2017 to March 2018 were collected. The patients were assessed for symptomatic distress with depression self-rating scale (SDS). Functional assessment of cancer therapy-breast (FACT-B) and the functional assessment of chronic illness therapy-spiritual well-being (FACIT-SP) were used to make the mental assessment. Multiple linear regression analysis was used to analyze the effect of symptom distress on the mental health of the patients.Results:The scores of symptom distress, mental health, FACT-B and FACIT-SP were (19.94±5.78), (50.68±10.64), (16.85±4.75), (33.83±8.33), respectively. Multivariate analysis showed that mental health score of the patients with symptom distress > 18 scores was reduced by 5.15 points ( P=0.01) compared with the patients with symptom distress≤18 scores. Compared with the patients with annual household income < 50 000 yuan, the mental health score of patients with annual household income of 50 000-79 000 yuan was increased by 9.46 points ( P < 0.01), and the mental health score of patients with annual family income ≥ 80 000 yuan was increased by 5.92 points ( P < 0.01); compared with the patients in phase I, the mental health score of the patients in phaseⅡwas decreased by 2.62 points ( P=0.02), and the mental health score of the patients in phase Ⅲ was decreased by 4.98 points ( P < 0.01). Conclusions:Symptom distress is an independent risk factor for affecting mental health of breast cancer patients. Solving symptom distress of patients can improve mental health status of the patients.