Fifty whitespotted bamboo sharks (Chiloscyllium playgiosum) of both sexes were used to establish a large capacity variable domain of the new antigen receptor (VNAR) library with a total capacity of over 10⁹ colony-forming units (CFU). It was applied to screen VNARs against human serum albumin (HSA) and human transcription factor EB (TFEB), respectively. Meanwhile, VNAR libraries specific to HSA and TFEB with capacities above 10⁸ CFU were obtained following conventional immunization. These two approaches were systematically studied in terms of VNAR yield and composition. By comparing the VNAR sequences obtained from naïve and antigen-immunized libraries, we found that the complementary-determining region 3 (CDR3) of the former differs in composition from that of the latter. It shares a higher degree of homology with the naïve library. Meanwhile, the binding efficiency assessed by ELISA is also different between the naïve and antigen-immunized libraries. The binding of VNARs from the TFEB-immunized library appeared to surpass that observed with the naïve libraries, whereas the performance of VNARs from the HSA-immunized library indicated that both the immunized and naïve libraries for HSA had positive binding responses in polyclonal and monoclonal ELISA. The results are useful to develop novel diagnostic and therapeutic products based on shark VNARs.

OBJECTIVE: To investigate the current status of early pain and agitation management in critically ill patients in Guizhou Province. METHODS: A retrospective study was performed using data collected from a quality control activity conducted between April and June 2021 in non-provincial public hospitals with general intensive care unit (ICU) in Guizhou Province. Hospital-level data included hospital name and grade, ICU staffing, and number of ICU beds. Patient-level data included characteristics of patients treated in the general ICU on the day of the survey (e.g., age, sex, primary diagnosis), as well as pain and agitation assessments and the types of analgesic and sedative medications administered within 24 hours of ICU admission. RESULTS: A total of 947 critically ill ICU patients from 145 hospitals were included, among which 104 were secondary-level hospitals and 41 were tertiary-level hospitals. Within 24 hours of ICU admission, 312 (32.9%) critically ill patients received pain assessments, and 277 (29.3%) received agitation assessments. Among the pain assessment tools, the critical care pain observation tool (CPOT) was used in 44.2% (138/312) of critically ill ICU patients, with a significantly higher usage rate in tertiary hospitals compared to secondary hospitals [52.3% (69/132) vs. 38.3% (69/180), P < 0.05]. The Richmond agitation-sedation scale (RASS) was used in 93.8% (260/277) of critically ill ICU patients for agitation assessment, with no significant difference between hospital levels. Among the 947 critically ill patients, 592 (62.5%) received intravenous analgesics within 24 hours, with remifentanil being the most commonly used [42.9% (254/592)]; 510 (53.9%) received intravenous sedatives, with midazolam being the most frequently used [60.8% (310/510)]. Mechanical ventilation data were available for 932 critically ill patients, of whom 579 (62.1%) received mechanical ventilation and 353 (37.9%) did not. Compared with non-ventilated patients, ventilated patients had significantly higher rates of analgesic and sedative use [analgesics: 77.9% (451/579) vs. 38.8% (137/353); sedatives: 71.8% (416/579) vs. 25.8% (91/353); both P < 0.05]. In terms of analgesic selection, ventilated patients were more likely to receive strong opioids than non-ventilated patients [85.8% (95/137) vs. 69.3% (387/451), P < 0.05]. For sedatives, ventilated patients preferred midazolam [66.6% (277/416)], whereas non-ventilated patients more often received dexmedetomidine [45.1 (41/91)]. Blood pressure within 24 hours of ICU admission were available for 822 critically ill patients, of whom 245 (29.8%) had hypotension and 577 (70.2%) did not. Compared with non-hypotensive patients, hypotensive patients had significantly higher rates of analgesic and sedative use [analgesics: 74.7% (183/245) vs. 59.8% (345/577); sedatives: 65.7% (161/245) vs. 51.3% (296/577); both P < 0.05], but there was no significant difference in the choice of analgesic or sedative agents between the two groups. CONCLUSIONS The proportion of critically ill ICU patients in Guizhou Province who received standardized pain and agitation assessments was relatively low. The most commonly used assessment tools were CPOT and RASS, while remifentanil and midazolam were the most frequently used analgesic and sedative agents, respectively. Secondary-level hospitals had a lower rate of using standardized pain assessment tools compared to tertiary-level hospitals. Mechanical ventilation and hypotension were associated with the use of analgesic and sedative medications.
Humans ; Critical Illness ; Intensive Care Units ; Analgesics/therapeutic use* ; Hypnotics and Sedatives/therapeutic use* ; Retrospective Studies ; China ; Pain Measurement ; Pain Management ; Female ; Male ; Critical Care ; Middle Aged

OBJECTIVE: To explore the timing of weaning extracorporeal membrane oxygenation (ECMO) and analyze the risk factors that affect survival outcomes before weaning. METHODS: A retrospective case-control study was conducted. Patients who received ECMO treatment and were weaned according to physicians' orders at the Second Xiangya Hospital of Central South University from January 2020 to June 2024 were enrolled as the study subjects. The general information, underlying diseases, indications and processes of ECMO, vital signs and arterial blood gas analysis 1 hour before weaning test, and biochemical indicators 24 hours before weaning test were collected through the hospital electronic medical record system. The primary outcome measure was the hospital mortality. The variables with P < 0.1 in univariate analysis and correlation analysis were included into binary Logistic regression analysis to identify risk factors. A nomogram model was constructed to predict the risk of weaning death in patients with ECMO, and receiver operator characteristic curve (ROC curve) and calibration curve were drawn to evaluate the model. Decision curve analysis (DCA) was used to evaluate the clinical net benefit rate of the model. RESULTS: A total of 32 ECMO patients were included, among whom 10 received veno-arterial ECMO (VA-ECMO) and 22 received veno-venous ECMO (VV-ECMO). During the hospitalization period, 23 patients survived, while 9 died. The time from mechanical ventilation to ECMO activation in the death group was significantly longer than that in the survival group, and the time from ECMO cessation to discharge was significantly shorter than that in the survival group. The levels of diastolic blood pressure (DBP) and albumin (Alb) before weaning were significantly lower than those in the survival group, and the level of procalcitonin (PCT) was significantly higher than that in the survival group (all P < 0.05). Spearman correlation analysis showed that DBP, PCT, Alb, and thrombin time (TT) were correlated with the weaning outcomes of ECMO patients (r values were -0.450, 0.373, -0.376, -0.346, all P < 0.1). Binary Logistic regression analysis showed that the final indicators entering the regression equation included DBP [odds ratio (OR) = 0.864, 95% confidence interval (95%CI) was 0.756-0.982], PCT (OR = 1.157, 95%CI was 0.679-1.973), and TT (OR = 0.852, 95%CI was 0.693-1.049), and a nomogram model was constructed to predict the weaning outcomes of ECMO patients. ROC curve analysis showed that the area under the curve (AUC) of the nomogram model for predicting the weaning outcome of ECMO patients was 0.831, with a sensitivity of 77.8% and a specificity of 65.2%. Its predictive value was better than that of single indicators DBP, PCT, and TT (AUC of 0.787, 0.739, and 0.722, respectively). The calibration curve showed that the prediction probability of the model was in good consistency with the actual observed results, the Hosmer-Lemeshow goodness of fit test showed that, χ ² = 8.3521, P = 0.400, indicating that the model fits well. DCA showed that across risk threshold of 0-0.8, the net benefit rate was greater than 0, which was significantly better than that of single indicator. CONCLUSIONS The nomogram model constructed with DBP, PCT, and TT has certain predictive value for the weaning outcomes of ECMO patients and can be used as a screening indicator for ECMO weaning timing.
Humans ; Extracorporeal Membrane Oxygenation ; Retrospective Studies ; Risk Factors ; Case-Control Studies ; Hospital Mortality ; Male ; Female ; Nomograms ; Logistic Models ; ROC Curve ; Middle Aged ; Adult ; Ventilator Weaning ; Time Factors

OBJECTIVE: To identify the risk factors related to the timing of patients receiving extracorporeal membrane oxygenation (ECMO) initiation and construct a risk prediction model for ECMO initiation timing. METHODS: Patients who received ECMO admitted to the Second Xiangya Hospital of Central South University from January 2020 to January 2024 were retrospectively collected. The case data mainly included physiological and biochemical indicators 1 hour before ECMO initiation. According to the outcome of the patients, they were divided into survival group and death group. Univariate and multivariate Logistic regression analysis were used to analyze the predictors of mortality risk in patients with ECMO, and a nomogram prediction model was constructed. The discrimination, calibration accuracy, and goodness of the model were evaluated by the receiver operator characteristic curve (ROC curve), calibration curve, and the Hosmer-Lemeshow test, respectively. Decision curve analysis (DCA) evaluated the clinical net benefit rate of the model. RESULTS: A total of 81 ECMO patients were included, including 59 males and 22 females; age range from 16 to 61 years old, with a median age of 56.0 (39.5, 61.5) years old; 20 patients received veno-arterial (V-A) ECMO, and 61 patients received veno-venous (V-V) ECMO; 23 patients ultimately survived and 58 patients died. Univariate analysis showed that age, blood urea nitrogen, serum creatinine, D-dimer, arterial blood carbon dioxide partial pressure, and prothrombin time of the death group were all higher than those of the survival group, while albumin was slightly lower than that of the survival group. There was a statistically significant difference in the direct cause of ECMO initiation between the two groups. Multivariate Logistic regression analysis showed that age [odds ratio (OR) = 1.069, 95% confidence interval (95%CI) was 1.015-1.125, P = 0.012], direct cause of ECMO initiation [with heart failure as the reference, return of spontaneous circulation (ROSC) after cardiopulmonary support (OR = 30.672, 95%CI was 1.265-743.638, P = 0.035), novel coronavirus infection (OR = 8.666, 95%CI was 0.818-91.761, P = 0.073), other severe pneumonia (OR = 4.997, 95%CI was 0.558-44.765, P = 0.150)], pre-ECMO serum creatinine (OR = 1.008, 95%CI was 1.000-1.016, P = 0.044), prothrombin time (OR = 1.078, 95%CI was 0.948-1.226, P = 0.252), and D-dimer (OR = 1.135, 95%CI was 1.047-1.231, P = 0.002) were entered into the final regression equation. A nomogram prediction model was developed based on these five factors. The area under the ROC curve (AUC) of the model was 0.889 (95%CI was 0.819-0.959), higher than the AUC of the sequential organ failure assessment (SOFA; AUC = 0.604, 95%CI was 0.467-0.742). The calibration curve showed good consistency between the model predictions and the observed results. The Hosmer-Lemeshow goodness-of-fit test showed that χ ² = 4.668, P = 0.792. DCA analysis showed that when the risk threshold was 0-0.8, the net benefit rate was greater than 0, which was significantly better than that of SOFA score. CONCLUSIONS The risk prediction model for the timing of ECMO initiation, constructed using five factors (age, direct cause of ECMO initiation, thrombin time, serum creatinine, and D-dimer), demonstrated good discrimination and calibration. It can serve as a pre-initiation assessment tool to identify and predict post-initiation mortality risk in ECMO patients.
Humans ; Extracorporeal Membrane Oxygenation ; Middle Aged ; Male ; Female ; Retrospective Studies ; Adult ; Risk Factors ; Adolescent ; Young Adult ; Logistic Models ; Nomograms ; ROC Curve ; Time Factors ; Risk Assessment

OBJECTIVE: To explore the incidence, clinical features, genetic variant characteristics and prognosis of Glutaric acidemia type I (GA1) among neonates from Henan Province. METHODS: A total of 814 625 neonates undergoing screening for inherited metabolic diseases by tandem mass spectrometry (MS/MS) at the Third Affiliated Hospital of Zhengzhou University from January 2016 to December 2022 were selected as the study subjects. A retrospective method was adopted to collect the clinical data of the patients. Whole exome sequencing was carried out to detect GCDH gene variants in individuals with positive results by GA1 newborn screening, and Sanger sequencing was used to verify the candidate variants. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the pathogenicity of candidate variants was rated. This study was approved by the Medical Ethics Committee of the Hospital (Ethics Number: 2019 Medical Ethics Review No. 67). RESULTS: Eight cases of GA1 were diagnosed among the 814 625 neonates. Blood glutaryl carnitine (C5DC) and urine glutaric acid (GA) levels of the 8 children were higher than the normal reference values. In total 12 variants were detected, all of which were missense variants. c.1064G>A (p.Arg355His) was the most common one, accounting for 21.4% (3/14). Three GCDH gene variants, including 1297G>C (p.Ala433Pro), c.467G>A (p.Gly156Asp) and c.1125T>G (p.Cys375Trp), were previously unreported. REVEL software analysis predicted that all of the three variants were harmful. 3D protein structure modeling indicated that the three variants may cause amino acid residue alterations, and c.1297G>C (p.Ala433Pro) and c.1125T>G (p.Cys375Trp) may result in increase in hydrogen bonds and affect the function of GCDH protein. By December 2023, one of the eight children had deceased, and another child had severe clinical symptoms with poor prognosis. Six children had a good prognosis, of which two had mild motor development delay and four had normal development without clinical symptoms. CONCLUSION The incidence of GA1 in newborns screened by MS/MS in Henan Province is 1/101 828, and the carrier rate of pathogenic GCDH variants is 1/160. The c.1064G>A (p.Arg355His) may be the hotspot variant of the GCDH gene among children with GA1 in Henan. Discovery of the three novel variants has enriched the mutational spectrum of the GCDH gene and provide a basis for the early diagnosis, treatment, prognosis and genetic counseling of this disease.
Humans ; Amino Acid Metabolism, Inborn Errors/epidemiology* ; Glutaryl-CoA Dehydrogenase/chemistry* ; Infant, Newborn ; Female ; Neonatal Screening/methods* ; Male ; Brain Diseases, Metabolic/epidemiology* ; China/epidemiology* ; Retrospective Studies ; Mutation ; Genetic Variation ; Glutarates

OBJECTIVE: To evaluate the differential expression of RNA in blood monocytes in patients with Juvenile idiopathic arthritis (JIA) treated with TNF antagonists (TNFi), and to explore the effect and mechanism of gene expression on the efficacy of JIA. METHODS: A total of 29 children with JIA treated with methotrexate (MTX) and TNFi in Guangzhou Women and Children's Medical Center of Guangzhou Medical University from April 2021 to November 2023 were enrolled. After 6 months, the children were divided into two groups according to the treatment effect, i.e., 13 cases in the ineffective group and 16 cases in the effective group, the peripheral blood of the children was collected, the blood mononuclear cells were isolated for transcriptome sequencing, the differentially expressed genes between the groups were analyzed, the signaling pathways and metabolic pathways related to the efficacy of TNFi were analyzed by GO and KEGG enrichment, and the mechanism related to the efficacy of TNFi was explored. This study was approved by Medical Ethics Committee of the Guangzhou Women and Children's Medical Center of Guangzhou Medical University (Ethics No.: 2023-330B00). RESULTS: There was a statistically significant difference in the gender and age distribution between the two groups of children (P < 0.05), while no statistically significant differences were observed in disease duration, rheumatoid antibody levels, or JIA subtypes (P > 0.05). After sequencing data quality control and comparison of reference genomes, a total of 18 523 protein-coding genes were identified in all children's samples. A total of 705 differentially expressed genes (DEGs) were identified between the effective group and the invalid group through differential analysis, of which 579 were up-regulated in the effective group and 126 in the inactive group. GO function and KEGG pathway enrichment analysis showed that DEG was significantly enriched in 55 GO entries and 32 KEGG metabolic pathways, which were mainly related to IL-1β production and regulation, cytokine production and regulation, cytokine-cytokine receptor interaction, immune response regulation, and Toll-like receptor signaling pathway. CONCLUSION DEG between the effective and ineffective groups of TNFi treatment may be involved in the biological processes such as cytokine production and regulation, cytokine-receptor interaction, and immune response regulation, which will be helpful to predict the efficacy and prognosis of TNFi treatment for JIA.
Humans ; Arthritis, Juvenile/blood* ; Female ; Male ; Child ; Methotrexate/therapeutic use* ; Child, Preschool ; Tumor Necrosis Factor-alpha/antagonists & inhibitors* ; Transcriptome ; Adolescent ; RNA/genetics* ; Signal Transduction ; Gene Expression Profiling

Objective:To explore the prevalence, clinical features, genetic characteristics and prognosis of Citrin deficiency in Henan province of China.Methods:A total of 986 565 neonates screened by tandem mass spectrometry at the Third Affiliated Hospital of Zhengzhou University from January 2013 to December 2021 were retrospectively analyzed. Analysis of SLC25A13 gene variants and parental verification were carried out for neonates suspected for Citrin deficiency by next-generation sequencing. The clinical, biochemical and genetic characteristics of Citrin deficiency patients were integrated to guide the diet treatment and follow up the growth and development. Paired- t test was used to compare the amino acid levels in the peripheral blood samples before and after the treatment. Results:Nine cases of Citrin deficiency were diagnosed among the 986 565 neonates. Specific elevation of citrulline was observed in all of the 9 cases. Six variants were detected by genetic sequencing, among which c. 852_855delTATG, c. 615+ 5G>A, c. 550C>T and IVS16ins3kb were known pathogenic variants, whilst c. 1111_1112delAT and c. 837T>A were unreported previously. The detection rate for c. 852_855delTATG was the highest (61.6%, 11/18), followed by IVS16ins3kb (16.7%, 3/18). The clinical symptoms of all patients were relieved after the treatment, and the blood amino acid profile and biochemical parameters were significantly improved by gradually falling within the normal range. By June 2022, all patients had shown a good prognosis.Conclusion:The prevalence of Citrin deficiency among neonates from Henan Province by tandem mass spectrometry is 1/109 618, and the carrier rate for the pathogenic variants of the SLC25A13 gene was 1/166. The c. 852_855delTATG may be a hot spot variant among the patients. Discovery of the novel variants has enriched the mutational spectrum of the SLC25A13 gene. Above results have provided a basis for the early diagnosis, treatment, prognosis and genetic counseling for the affected families.

Objective:To explore the differences of microenviroment between peri-implant tissue and oral mucosal tissue.Methods:The gene chip data GSE43744 was downloaded from the GEO database,bioinformatics tools were used to analyze the differentially ex-pressed genes between the peri-implant tissue and normal oral mucosal tissue in rat.Results:1315 differentially expressed important genes,including 797 upregulated genes and 518 downregulated genes,were screened out.Gene enrichment analysis showed that com-pared with normal oral mucosal tissue,the gene expression of innate immune activity,cell activation,inflammatory response,and func-tional expression related to external and bacterial stimuli in peri-implant tissue were significantly upregulated,while that of extracellular matrix tissue,adhesion,extracellular matrix polysaccharides,response to mechanical stimuli and response to toxic substances was sig-nificantly downregulated.Meanwhile,multiple molecular functions and biological pathways related to T cells were highly expressed,which may play an important role in the peri-implant microenvironment.In addition,PPI network was constructed,and screened 7 core genes including FCER1G,TYROBP,PTPRC,ITGB2,AIF1,EMR1 and RAC2,which may be target genes for studying peri-implant microenvironment.Conclusion:There is a significant difference of microenvironment characteristics between peri-implant tissue and o-ral mucosa.The target genes screened using PPI network may be the key to future research on the peri-implant microenvironment.

Purpose/Significance To explore the research status,research hotspots and frontiers in the field of clinical decision sup-port based on electronic medical records(EMR).Method/Process The bibliometric method and CiteSpace 6.2.R2 software are used to draw scientific knowledge graph of country/region distribution,author cooperation,institutional cooperation,keyword co-occurrence and clustering for visualized comparative analysis.Python is used for clustering hotspot mining and analysis.Result/Conclusion The field of clinical decision support based on EMR data shows a rapid development trend,with the United States and China as the main research countries and strong cooperation between domestic and foreign institutions.The keywords mainly involve EMR,artificial intelligence(AI),etc.

Seven novel linear polyketides,talaketides A-G(1-7),were isolated from the rice media cultures of the mangrove sed-iment-derived fungus Talaromyces sp.SCSIO 41027.Among these,talaketides A-E(1-5)represented unprecedented unsaturated lin-ear polyketides with an epoxy ring structure.The structures,including absolute configurations of these compounds,were elucidated through detailed analyses of nuclear magnetic resonance(NMR)and high-resolution mass spectrometry(HR-MS)data,as well as elec-tronic custom distributors(ECD)calculations.In the cytotoxicity screening against prostate cancer cell lines,talaketide E(5)demon-strated a dose-dependent inhibitory effect on prostate cancer PC-3 cell lines,with an IC50 value of 14.44 μmol·L-1.Moreover,com-pound 5 significantly inhibited the cloning formation of PC-3 cell lines and arrested the cell cycle in S-phase,ultimately inducing ap-optosis.These findings indicate that compound 5 may serve as a promising lead compound for the development of a potential treat-ment for prostate cancer.