Objective: To propose an improved method for constructing the internal quality control (IQC) framework for ELISA assays and validate its efficacy by statistically analyzing IQC data from nine blood center laboratories. Methods: 1) IQC data was collected from nine blood centers and analyzed using a domestic HBsAg ELISA detection kit as an example. 2) Differences between IQC values across batches within Blood Center 1 were assessed. 3) Statistical analyses were performed on batch usage, number of batches used, days of use, number of QC points, batch-specific means, and coefficients of variation (CV) across all nine centers. 4) Using the improved construction method for IQC framework, provisional and permanent frames were established for batches within Blood Center 1 and Blood Center 9, followed by outlier determination. Results: 1) Statistically significant differences were observed in IQC data between batches within Blood Center 1 (P<0.01). It is recommended that both the control material/reagents and the control chart framework be replaced simultaneously. 2) There were substantial differences among 9 blood centers regarding the control material/reagent lot numbers used, the number of QC runs per batch, and the QC values for identical lots. Therefore, individual laboratories should establish their own IQC chart frameworks. 3) The improved IQC framework construction method for ELISA assays is as follows: provisional frames are established via frame-shifting, using the pre-experimental mean and cumulative coefficient of variation (CV) from the preceding batch. For batches used >20 days with >20 QC points, permanent frames are constructed by aggregating in-control data accumulated over ≥20 days with ≥20 points to calculate cumulative mean and standard deviation. The provisional and permanent frames constructed by this method identified all 26 extreme outliers across Blood Centers 1 and 9 as out-of-control. Among the 218 general outliers, 10 were classified as normal by the provisional frames, while the remainder were designated as warnings or out-of-control. This method effectively monitors assay stability. Conclusion: Based on the statistical analysis of IQC practices across blood centers of varying scales, combined with the inherent characteristics of ELISA assays and the batch-to-batch instability of reagents/QC materials, it is recommended to reconstruct QC charts upon lot changes. The proposed method—utilizing frame-shifting for provisional frames and establishing permanent frames based on cumulative data—is applicable to blood center laboratories of differing sizes and effectively monitors the stability of the ELISA assay process.

Objective: To analyze the epidemiological characteristics and incidence trends of pulmonary tuberculosis (TB) in children aged 0-14 years in Shaanxi Province from 2010 to 2024, so as to provide a reference for optimizing child TB prevention and control strategies. Methods: Data on pulmonary TB cases in children aged 0-14 years and demographic information in Shaanxi Province from 2010 to 2024 were collected from Surveillance and Reporting Management System with Disease Prevention and Control Information Management System under the National Health Security Informatization Project Disease Prevention and Control Information System. A Joinpoint regression model was established to analyze the temporal, spatial, and population distribution trends of child pulmonary TB incidence. Results: A total of 2 954 cases of pulmonary TB in children aged 0-14 years were reported in Shaanxi Province from 2010 to 2024, accounting for 0.97% of all TB cases in the general population. The average annual reported incidence rate in children was 3.32 per 100 000. Among these cases, 804 were pathogenetically positive, showing a increasing trend ( χ 2 trend =420.94, P < 0.01 ). The overall reported incidence rate of pulmonary TB in children aged 0-14 years in Shaanxi Province showed a decreasing trend, dropping from 5.35 per 100 000 in 2010 to 2.41 per 100 000 in 2024. Joinpoint regression analysis identified three distinct phases for the reported incidence rate of TB:a rapid decline from 2010 to 2013 (APC=-20.02%, 95% CI = -33.64% to -10.42%), a slight increase from 2013 to 2017 (APC=11.18%, 95% CI =3.07%-24.17%) and a slight decline again from 2017 to 2024 (APC= -7.27 %, 95% CI =-12.73% to -4.30%) (all P <0.01). Among children aged 0-14 years, the age group with the highest average annual reported incidence rate was 10-14 years (8.02 per 100 000), followed by 5-9 years (1.44 per 100 000), and 0-4 years had the lowest rate (0.95 per 100 000). The difference in reported incidence rates among the three age groups was statistically significant ( χ 2= 51.91, P <0.01). The average annual reported incidence rate of TB was 3.25 per 100 000 in boys and 3.39 per 100 000 in girls, with no statistically significant difference ( χ 2=2.01, P >0.05). There was no obvious periodic variation in the annual case reporting. Among all cities in Shaanxi Province, Ankang City had the highest average annual reported incidence rate (5.16 per 100 000). Conclusions From 2010 to 2024, the reported incidence rate of pulmonary TB in children aged 0-14 years in Shaanxi Province showed an overall decreasing trend. However, it is still necessary to strengthen active surveillance, implement targeted measures in high incidence areas such as Ankang City, and maintain continuous attention to child TB prevention and control.

Objective:To prepare a novel ginseng polypeptide thermosensitive hydrogel,and to investigate its repair effect on heat-induced skin injury in the rats and explore the underlying mechanisms.Methods:Thermosensitive hydrogels were formulated using Pluronic F127 and β-sodium glycerophosphate(β-GP),and their phase transition temperatures,spatial structures,elemental compositions,and water retention capacities were evaluated.The rat models of heat-induced skin injury were established and the model rats were divided into PBS group,Gel group,and ginseng polypeptide gel(GP-Gel)group.After 11 d of treatment,the morphological changes of wound and collagen deposition in the wound of the rats in various groups were observed by HE and Masson staining.Immunohistochemistry was used to detect the expression levels of α-smooth muscle actin(α-SMA),connective tissue growth factor(CTGF),basic fibroblast growth factor(bFGF),proliferating cell nuclear antigen(PCNA),cell proliferation marker Ki67,epidermal growth factor(EGF),CD31,vascular endothelial growth factor(VEGF),P50 and P65 proteins in the skin wound tissue of the rats in various groups.Western blotting method was used to detect the expression levels of Toll-like receptor 4(TLR4)in the skin wound tissue of the rats in various groups.ELISA method was used to measure the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-15(IL-15),and interleukin-10(IL-10)in the serum of the rats in various groups.Results:Compared with PBS and Gel groups,the wound area of the rats in GP-Gel group was reduced(P＜0.01),the expression levels of PCNA,Ki67,EGF,CD31,VEGF,α-SMA,and CTGF proteins in the skin wound tissue were increased(P＜0.05 or P＜0.01),and the expression levels of P65 and TLR4 proteins were decreased(P＜0.01);the level of anti-inflammatory factor IL-10 in serum was increased(P＜0.01),while the levels of pro-inflammatory factors TNF-α,IL-1β,IL-6 and IL-15 were decreased(P＜0.05 or P＜0.01).Conclusion:The ginseng polypeptide thermosensitive hydrogel promotes the repair of heat-induced skin injury by enhancing cell proliferation,collagen synthesis,angiogenesis,and reducing inflammatory responses.

Objective:To develop and validate a novel PET tracer, N-cyclohexyl-4-((2, 4-dichlorophenyl)(4-(fluoro- 18F)phenyl)methyl)piperazine-1-carboxamide ( 18F-SQKJ-2), targeting cannabinoid type 1 (CB1) receptors for diagnosing psychiatric disorders associated with fear memory. Methods:18F-SQKJ-2 was prepared using a nucleophilic substitution radiochemical synthesis method. For the CB1 receptor blocking experiment, 7 ICR mice were divided into blocking group ( n=4; rimonabant for blocking treatment) and control group 1 ( n=3; no rimonabant blocking treatment). The affinity and specificity of 18F-SQKJ-2 for CB1 receptors were analyzed based on the differences in 18F-SQKJ-2 uptake (percentage injected dose per gram of tissue, %ID/g) by various organs between two groups. The metabolic stability of 18F-SQKJ-2 in vitro was studied using animal tissue homogenates. Ten C57 mice were used to establish fear memory mouse models (fear group, n=6; control group 2, n=4), and the percentage of freezing time was compared between 2 groups. MicroPET scans were used to detect the intracranial distribution of 18F-SQKJ-2, and the relative uptake in each brain region compared to total brain uptake was calculated. Statistical analysis was conducted to compare the differences in CB1 receptor relative total brain uptake in fear-related brain regions between 2 groups. Independent-sample t test and Mann-Whitney U test were used to analyze the data. Results:18F-SQKJ-2 was successfully synthesized with a radiochemical purity ≥98.0% and a corrected radioactive yield of (12.3±6.0)%( n=4). In vitro metabolic stability experiments showed that 18F-SQKJ-2 was basically stable in the liver, blood, and brain within 60min. The CB1 receptor blocking experiment demonstrated that the uptake of 18F-SQKJ-2 in the brains of mice in blocking group was significantly lower than that in control group 1 ((0.95±0.28) vs (3.44±1.16) %ID/g; t=-3.57, P=0.023). The percentage of freezing time in fear group was significantly higher than that in control group 2 (43.28%(39.46%, 52.93%) vs 2.74%(1.52%, 4.85%); Z=-2.45, P=0.010). 18F-SQKJ-2 microPET imaging showed that the uptake of 18F-SQKJ-2 in the cerebral cortex of mice in fear group was significantly increased compared with that in control group 2 ((5.83±0.47)% vs (5.00±0.52)%; t=2.42, P=0.046). Conclusion:18F-SQKJ-2 is successfully prepared with acceptable radiochemical purity and metabolic stability, demonstrating potential for visualizing and quantifying fear memory.

Objective:To assess the clinical value of digital PCR (dPCR) for the prenatal diagnosis of common fetal aneuploidies.Methods:A dPCR-based assay was developed for detecting trisomies 21, 18, and 13. A retrospective analysis was carried out on 173 amniotic fluid samples collected by the Prenatal Diagnosis Center of Taizhou Hospital between January 2017 and December 2023. By using chromosomal karyotyping as the gold standard, the diagnostic performance of the multiplex dPCR system was evaluated in a double-blind manner. This study has been approved by the Ethics Committee of Taizhou Hospital (Ethics No. K20250339).Results:Chromosomal karyotyping has identified 59 cases of trisomy 21, 5 cases of trisomy 18, 2 cases of trisomy 13, 6 cases with chromosomal structural abnormalities or mosaicisms, and 101 cases with a normal karyotype. The dPCR results ( Z-score cutoff = 4.0, CI = 99.997%) showed full concordance with karyotyping (sensitivity = 100%, specificity = 100%, Kappa = 1). Among the 6 structurally abnormal or mosaicism samples, dPCR has accurately detected 4 cases, but mis-classified 2 cases of trisomy 21 with very low-level mosaicisms (3.3%, 6.9%, respectively) as normal. Conclusion:The established multiplex dPCR system demonstrated high diagnostic accuracy for common chromosomal aneuploidies, with results available within 24 hours. It can serve as an efficient supplementary tool to conventional chromosomal karyotyping, providing reliable support for time-sensitive clinical decision-making in prenatal diagnosis.

OBJECTIVE: To assess the clinical value of digital PCR (dPCR) for the prenatal diagnosis of common fetal aneuploidies. METHODS: A dPCR-based assay was developed for detecting trisomies 21, 18, and 13. A retrospective analysis was carried out on 173 amniotic fluid samples collected by the Prenatal Diagnosis Center of Taizhou Hospital between January 2017 and December 2023. By using chromosomal karyotyping as the gold standard, the diagnostic performance of the multiplex dPCR system was evaluated in a double-blind manner. This study has been approved by the Ethics Committee of Taizhou Hospital (Ethics No. K20250339). RESULTS: Chromosomal karyotyping has identified 59 cases of trisomy 21, 5 cases of trisomy 18, 2 cases of trisomy 13, 6 cases with chromosomal structural abnormalities or mosaicisms, and 101 cases with a normal karyotype. The dPCR results (Z-score cutoff = 4.0, CI = 99.997%) showed full concordance with karyotyping (sensitivity = 100%, specificity = 100%, Kappa = 1). Among the 6 structurally abnormal or mosaicism samples, dPCR has accurately detected 4 cases, but mis-classified 2 cases of trisomy 21 with very low-level mosaicisms (3.3%, 6.9%, respectively) as normal. CONCLUSION The established multiplex dPCR system demonstrated high diagnostic accuracy for common chromosomal aneuploidies, with results available within 24 hours. It can serve as an efficient supplementary tool to conventional chromosomal karyotyping, providing reliable support for time-sensitive clinical decision-making in prenatal diagnosis.
Humans ; Female ; Pregnancy ; Aneuploidy ; Prenatal Diagnosis/methods* ; Karyotyping ; Retrospective Studies ; Polymerase Chain Reaction/methods* ; Chromosome Disorders/genetics* ; Adult ; Trisomy 13 Syndrome/diagnosis* ; Trisomy 18 Syndrome/genetics* ; Down Syndrome/genetics*

AIM:To investigate the protective ef-fect of osthole(Ost)on APAP-induced liver injury in mice and its molecular mechanism.METHODS:We established the APAP-induced liver injury model in mice,and Ost was used to intervene.The expres-sion of AST,ALT,SOD,ROS,MDA,LDH,GSH-PX in mice plasma were detected by biochemical meth-od.HE staining was used to observe the changes of liver tissue structure.Immunofluorescence assay was used to detect the expression of Tif1γ and Smad4 in liver tissue.The mRNA expression of IL-1β,IL-6,TNF-α,Smad4,and Tif1γ were detected by qRT-PCR.Western blot was applied to assess the protein expression of Smad2/3 and pSmad2/3 in liver tissue.RESULTS:Compared with the control group,the liver structure destruction and hepato-cyte death was increased,ALT,AST,ROS,MDA and LDH were increased,while SOD and GSH-PX were decreased,and the mRNA expressions of IL-1β,IL-6 and TNF-α were increased in the model group.Compared with the model group,the Ost interven-tion group had improved liver structure and de-creased liver cell death;decreased ALT,AST,ROS,MDA and LDH,increased SOD and GSH-PX,and de-creased expression of IL-1β,IL-6 and TNF-α mRNA.Compared with the control group,liver tissues of model mice showed increased expression of pS-mad2/3,Smad4 protein and Smad4 mRNA,and de-creased Tif1γ protein and mRNA.Compared with the model group,the liver tissues of the Ost inter-vention group showed decreased expression of pS-mad2/3,Smad4 protein and Smad4 mRNA,and in-creased expression of Tif1γ protein and mRNA.CONCLUSION:Ost can improve liver function,re-duce oxidative stress and inflammatory reaction,and protect hepatocyte damage induced by APAP in mice,which may be related to the up-regulation of Tif1γ and inhibition of TGF-β1/Smad signaling pathway.

Objective:To understand the spatiotemporal distribution of pulmonary tuberculosis (TB) in Shaanxi Province from 2015 to 2023, and provide reference for the prevention and control of pulmonary TB in Shaanxi.Methods:The registration data of etiologically positive pulmonary TB cases in Shaanxi from 2015 to 2023 were collected from the tuberculosis subsystem of Chinese Disease Control and Prevention Information System. Descriptive method was used to analyze the basic characteristics of the etiologically positive pulmonary TB cases. Linear trend χ2 test was used to analyze trends in registration rate and pathogen positive rate. Software SPSS 25.0 was used for statistical analysis. Software ArcGIS 10.8 was used for global spatial autocorrelation and hotspot analysis to explore spatial clustering of the etiologically positive pulmonary TB cases. Software SaTScan 10.0 was used for spatiotemporal scan statistics, and software ArcGIS 10.8 was used to visualize the spatiotemporal clustering. Results:A total of 64 148 cases of etiologically positive pulmonary TB were registered in Shaanxi from 2015 to 2023, with an average annual registration rate of 18.33/100 000. The registration rate and pathgen positive rate all showed upward trends from 2015 to 2023, and the differences were significant (the trend χ2=4 555.18 and 19 330.43, both P<0.001). Global spatial autocorrelation and hotspot analysis showed that the registration rate of etiologically positive pulmonary TB in Shaanxi from 2017 to 2023 showed a spatial clustering. The hotspots were mainly in Zhenba and Xixiang counties of Hanzhong, six counties (districts) of Ankang, and Yanchuan and Yanchang counties of Yan'an. The coldspots were mainly in parts of the Guanzhong area, including Baoji, Xi'an, and Xianyang. A total of 4 spatiotemporal clustering areas were explored by spatiotemporal scanning analysis (all P<0.001), in which the first-level clustering areas covered 17 counties (districts), mainly Zhenping, Ziyang, Zhenba, in southern Shaanxi from 2019 to 2022, the second-level clustering areas covered 6 counties (districts), mainly Yanchuan, Yanchang, Qingjian, in northern Shaanxi from 2018 to 2021, the third-level clustering areas covered 14 counties (districts), mainly Yanta, Chang'an, Jingyang, in Guanzhong area from 2018 to 2019, and the fourth-level clustering areas covered 10 counties (districts) from 2019 to 2021. Conclusions:The registration rate of labortory confirmed pulmonary TB cases in Shaanxi showed an upward trend, with obvious differences in spatiotemporal clustering distribution. The clustering areas were mainly in southern Shaanxi, such as Zhenba, Zhenping, Hanbin, Langao, Pingli, Xunyang, Ziyang counties, and northern Shaanxi, such as Yanchuan and Yanchang counties, as well as in capital city, Xi'an and the adjacent Guanzhong area. It is necessary to develope targeted measures according to local conditions for the improvement of pulmonary TB prevention and control strategies in Shaanxi.

BACKGROUND:Adolescent lumbar disc herniation is the main cause of low back pain in adolescents. At present,most of them are treated by conservativetreatment. When long-term non-surgical treatment attempts,surgery may be necessary to prevent further injury when the patient's symptoms are notsufficiently relieved or when the patient has symptoms of single nerve paralysis or compression of the cauda equina,it is very important to choose a suitable interbody fusion device for the surgical treatment of the patients.OBJECTIVE:To explore the effects of minimally invasive interbody fusion with different heights on lumbar biomechanics in patients with adolescent lumbar disc herniation.METHODS:CT scans of a 17-year-old male patient with adolescent lumbar disc herniation (L4-5 segment herniation) were collected. After the three-dimensional reconstruction of MIMICS,the interbody fusion device equal to and 3 mm higher than the intervertebral space was selected for analysis,so two expandablemixed material interbody fusion devices were designed and reconstructed. Fusion device L:11 mm high front,9 mm high posterior,9 mm wide,28 mmlong,and fusion device H:14 mm high front,11 mm high posterior,11 mm wide,28 mm long and the lumbar fusion device was modeled. The fusion deviceand lumbar spine model were optimized,inversely modeled,and then imported into ABAQUS,and finally the 3D model of lumbar fusion was obtained.The physiological activities of the human body were simulated,such as lumbar extension,forward bending,right bending,and left bending,to obtain thecorresponding stress contours. The biomechanical characteristics of the L4-5 vertebra under seven different working conditions were observed.RESULTS AND CONCLUSION:(1) The maximum stress of the two kinds of fuses was in the condition of forward bending and backward extension,the stress value of H fuses was (18.27±3.80) Mpa and (15.02±3.24) Mpa;the stress value of L fuses was (9.16±0.05) Mpa and (9.17±1.83) Mpa. The stress values of the end plate of the H-fusion in the extension station were (19.11±4.03) Mpa and (16.32±3.72) Mpa respectively. The stress values of the L-fusion end plate were (9.13±0.01) Mpa and (4.92±1.01) Mpa respectively. (2) The stress of H-type fusing end plate was higher than that of L-type fusing end plate except for L-5 end plate at neutral position (P<0.05). (3) Choosing an interbody fusion device with a height of more than 3 mm in the same intervertebral space has a more stable biomechanics.

Objective:To explore the changes of brain excitation/inhibition balance and gray matter volume (GMV) and their correlations with clinical features in benign childhood epilepsy with centrotemporal spikes (BECTS).Methods:A cross-sectional study was performed; 83 BECTS children enrolled from Department of Diagnostic Radiology, Jinling Hospital, Affiliated Hospital of Medical School, Nanjing University from January 2015 to January 2024 were selected as BECTS group. During the same period, 101 age- and gender-matched healthy children were recruited as healthy control group through advertisements in local primary schools. Data of conventional MRI and resting-state functional MRI (rs-fMRI) of the two groups were collected. Whole brain GMV was analyzed by voxel-based morphometry (VBM), and Hurst index was calculated based on time series data of blood oxygen level dependent (BOLD) signal of rs-fMRI. Correlations of GMV and Hurst index with disease duration and onset age in children with BECTS were explored by Pearson correlation analysis.Results:Compared with the healthy control group, the BECTS group had significantly increased GMV and decreased Hurst index in the bilateral Rolandic region ( P<0.05). Pearson correlation analysis showed that in the BECTS group, GMV in bilateral Rolandic region was negatively correlated with onset age ( r=-0.267, P=0.015) and positively correlated with disease course ( r=0.267, P=0.015); Hurst index in bilateral Rolandic region was positively correlated with onset age ( r=0.323, P=0.003) and negatively correlated with disease course ( r=-0.240, P=0.029); Hurst index was negatively correlated with GMV in bilateral Rolandic region ( r=-0.328, P=0.003). Conclusion:BECTS children have excitation/inhibition imbalance in epilepsy-related regions and cortical structural delay, and both of them are related to onset age and disease course.