ObjectiveTo investigate the role of DEAD-box helicase 3 X-linked （DDX3X） in immune-mediated liver injury （ILI）， and to clarify its mechanism by regulating endoplasmic reticulum stress （ERS）-dependent apoptotic pathway and its association with the clinical progression of hepatitis B. MethodsMice were given injection of concanavalin A （ConA） via the caudal vein to establish a model of ILI， PBS （control group） and different concentrations of ConA were injected into the tail vein of hepatocyte-specific DDX3X-knockout mice （DDX3X^ΔHep and DDX3X-flox mice （DDX3X^fl/fl）， respectively.. The log-rank survival analysis， measurement of the serum levels of aspartate aminotransferase （AST） and alanine aminotransferase （ALT）， and HE staining of liver tissue were performed to assess liver injury， and qRT-PCR and Western Blot were used to measure the mRNA and protein expression levels of glucose-regulated protein 78 （GRP78）， CCAAT/enhancer-binding protein homologous protein （CHOP）， and DDX3X in liver tissue. Intraperitoneal injection of 4-phenylbutyric acid （4-PBA， 100 mg/kg） was performed to inhibit ERS. Serum samples （n=30） and liver tissue samples （n=6） were collected from healthy controls， chronic hepatitis B （CHB） patients， and hepatitis B virus-associated liver failure （HBV-LF） patients； ELISA was used to measure the serum level of DDX3X， and qRT-PCR/Western Blot was used to analyze the expression of targets in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the control group of mice， the expression of DDX3X in the liver of mice induced by ConA was significantly increased after liver injury （P<0.05）， and hepatocyte-specific DDX3X knockout increased the 72-hour survival rate of mice by 55% （compared with 20% in the DDX3X^fl/fl group）， with significant reductions in the serum levels of ALT and AST （P<0.000 1） and the expression levels of the ERS markers GRP78 and CHOP （P<0.05）. After ERS was inhibited by 4-PBA， there was alleviation of liver injury （with reductions in ALT and AST， P <0.001） and a reduction in DDX3X expression （P<0.01）. The analysis of clinical samples showed that the mRNA and protein expression levels of liver DDX3X in CHB patients and HBV-LF patients were significantly higher than those in healthy controls （all P<0.01）， and there was a significant increase in the serum level of DDX3X in HBV-LF patients （P<0.000 1）. ConclusionDDX3X exacerbates ILI by regulating the ERS-dependent apoptotic pathway （GRP78/CHOP）， and its expression is associated with the progression of hepatitis B. Therefore， it can be used as a potential therapeutic target.

Objective To investigate the knowledge of tuberculin skin test(TST)among healthcare workers and provide evidence for improving the standardization of TST screening in primary healthcare staff.Methods A questionnaire survey was conducted among 248 licensed physicians or nurses who were qualified as licensed physicians or nurses and responsible for TST work from 27 districts/counties of Chongqing in 2023.The awareness of TST-related knowledge and its influencing factors were statistically analyzed.Results The average TST knowledge score of 248 healthcare workers was(78.3±10.6)points.The overall awareness rate was 78.9%(8 213/10 416),with specific rates as follows:65.4%(1 135/1 736)for tubercu-losis knowledge,87.3%(3 248/3 720)for TST general knowledge,53.4%(795/1 488)for TST principles,88.0%(1 964/2 232)for TST procedures,and 86.4%(1 071/1 240)for TST result interpretation.Nurses showed higher awareness rates than physicians and other staff(P＞0.05).Healthcare workers from medium-epidemic areas demonstrated significantly higher awareness rates than those from high-and low-epidemic are-as(P＜0.001).No statistically significant differences were observed in gender,age,occupation type,institu-tion type,or regional epidemic level between the qualified group and non-qualified group about TST-related knowl-edge(P＞0.05).Conclusion Healthcare workers exhibit incomplete mastery of TST-related knowledge.Strengthening TST-related knowledge training for standardizing TST implementation.

Microwave thermochemotherapy (MTC) has been applied to treat lip squamous cell carcinoma (LSCC), but a deeper understanding of its therapeutic mechanisms and molecular biology is needed. To address this, we used single-cell transcriptomics (scRNA-seq) and spatial transcriptomics (ST) to highlight the pivotal role of tumor-associated neutrophils (TANs) among tumor-infiltrating immune cells and their therapeutic response to MTC. MNDA⁺ TANs with anti-tumor activity (N1-phenotype) are found to be abundantly infiltrated by MTC with benefit of increased blood perfusion, and these TANs are characterized by enhanced cytotoxicity, ameliorated hypoxia, and upregulated IL1B, activating T&NK cells and fibroblasts via IL1B-IL1R. In this highly anti-tumor immunogenic and hypoxia-reversed microenvironment under MTC, fibroblasts accumulated in the tumor front (TF) can recruit N1-TANs via CXCL2-CXCR2 and clear N2-TANs (pro-tumor phenotype) via CXCL12-CXCR4, which results in the aggregation of N1-TANs and extracellular matrix (ECM) deposition. In addition, we construct an N1-TANs marker, MX2, which positively correlates with better prognosis in LSCC patients, and employ deep learning techniques to predict expression of MX2 from hematoxylin-eosin (H&E)-stained images so as to conveniently guide decision making in clinical practice. Collectively, our findings demonstrate that the N1-TANs/fibroblasts defense wall formed in response to MTC effectively combat LSCC.
Humans ; Neutrophils/metabolism* ; Single-Cell Analysis ; Lip Neoplasms/genetics* ; Hyperthermia, Induced/methods* ; Microwaves/therapeutic use* ; Transcriptome ; Carcinoma, Squamous Cell/immunology* ; Tumor Microenvironment

Objective To investigate the correlation between the levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and C-reactive protein(CRP)in the serum of full-term small-for-date infants and their growth restriction.Methods Pregnant women and their newborns who underwent routine check-ups at the Department of Obstetrics of Xingtai Central Hospital were enrolled.The mothers were admitted for delivery between January 2022 and December 2023.Newborns with a gestational age between 37 weeks and 41 weeks 6 days were included.A total of 83 newborns weighing<2 500 g at birth were included in the full-term small-for-date infant group,while 72 newborns weighing≥2 500 g at birth were included in the healthy control group.The maternal and neonatal serum levels of TNF-α,IL-6,and CPR were compared between the two groups.Logistic regression analysis was performed to screen for influencing factors.Receiver operating characteristic(ROC)curves were plotted to assess the predictive value of each influencing factor,and the optimal cutoff value,sensitivity,and specificity were derived subsequently.Results Compared with the healthy group,the full-term small-for-date infant group had elevated maternal and neonate serum levels of TNF-α,IL-6,and CPR(P<0.001).Maternal body mass index(BMI)(OR=0.428;95%CI,0.238-0.768;P=0.004),TNF-α levels(OR=2.133;95%CI,1.012-4.496;P=0.046),IL-6 levels(OR=1.218;95%CI,1.121-1.322;P<0.001),and CPR levels(OR=1.733;95%CI,1.312-2.288;P<0.001)were significantly associated with the incidence of full-term small-for-date infants(P<0.05).The area under the ROC curve(AUC)for maternal BMI and maternal serum TNF-α,IL-6,and CPR levels were 0.358(0.271-0.444),0.735(0.656-0.814),0.838(0.777-0.898),and 0.743(0.666-0.820),respectively.Among the 83 cases of full-term small-for-date infants,49 cases(59.04%)achieved satisfactory weight according to infant weight evaluation standards by the age of 6 months.Only birth weight(OR=1.004;95%CI,1.312-2.288;P<0.001)was identified as a significant influencing factor for satisfactory catch-up growth in full-term small-for-date infants.There was no significant association between the levels of inflammatory cytokines at birth and satisfactory catch-up growth at 6 months of age.Conclusion Maternal BMI and maternal and neonatal serum levels of TNF-α,IL-6,and CPR are all associated with the occurrence of growth restriction in full-term small-for-date infants.Measuring maternal serum levels of TNF-α,IL-6,and CPR may have value in predicting the occurrence of full-term small-for-date infants.However,no significant correlation is identified between the neonate serum levels of TNF-α,IL-6,and CRP and their growth catch-up at 6 months of age.

Objective To examine the association between lymphocyte subsets and renal clinicopathological characteristics as well as prognosis in patients with IgA nephropathy(IgAN).Methods The retrospective analysis included general clinical data and pathological examination results of IgAN patients diagnosed by renal biopsy at the Fourth Hospital of Hebei Medical University from January 2018 to January 2022.Correlation tests were conducted to examine the relationship between lymphocyte subsets and other significant clinicopathological parameters.The optimal cut-off value of CD4+T determined using the Youden index,and patients were grouped accordingly.Kaplan-Meier survival curves and Cox regression analyses were employed to compare the low and high CD4+T lymphocyte groups among IgAN patients,identifying factors influencing renal function progression.The endpoint event was defined as a decrease in estimated glomerular filtration rate(eGFR)of≥30%from baseline,progression to end-stage renal disease(ESRD)[eGFR<15 mL/(min·1.73 m2)or initiation of renal replacement therapy],or all-cause mortality.Results Low CD4+T lymphocytes were significantly positively correlated with blood IgA levels and the proportion of glomerular crescents in IgAN patients(all P<0.05).This study included a total of 53 IgAN patients,divided into two groups based on CD4+T lymphocyte counts:20 patients in the low CD4+T lymphocyte group and 33 patients in the high CD4+T lymphocyte group.In the low CD4+T lymphocyte group,there was a higher proportion of males and a lower proportion of glomerular crescents(P<0.05).Kaplan-Meier survival analysis revealed that patients with low CD4+lymphocytes had a significantly lower cumulative renal survival rate(Log-Rank test χ2=4.188,P=0.041).Cox regression analysis indicated that low CD4+lymphocytes were an independent risk factor for the progression of renal function decline in IgAN patients(HR=2.614,95%CI:1.006～6.788,P=0.048).Conclusions Patients with higher levels of CD4+T lymphocytes exhibit a lower risk of adverse renal outcomes.In contrast,patients with IgA nephropathy and low CD4+T lymphocyte counts tend to have poorer renal survival rates.

Objective To investigate the immediate effect of transcranial direct current stimulation(tDCS)combined with seated Tai-jiquan Yunshou training under different sequences on cerebral cortical activation in stroke patients with hemiple-gia.Methods From September to December,2024,14 stroke inpatients with hemiplegia were enrolled from the Fifth Hospi-tal of Xiamen.Based on the routine medication and rehabilitation,the patients were randomly assigned to a spe-cific intervention sequence,receiving three interventions in a fixed order:tDCS followed by Yunshou(S-Y group),Yunshou followed by tDCS(Y-S group),and simultaneous tDCS and Yunshou(Sim group).Functional near-infrared spectroscopy was used to detect oxyhemoglobin(HbO?)concentration in bilateral sensorimotor cor-tex,premotor and supplementary motor cortex,and prefrontal cortex before and after each intervention.Results Three patients dropped out.In all the regions of interest,inter-group effects were significant in HbO? concentra-tions(F>3.697,P<0.05),and interaction effects were significant in some brain regions(F>3.276,P<0.05).Post-hoc test showed a general trend of Sim Group≥S-Y Group>Y-S Group(P<0.05),although some varia-tions existed across different brain regions.Conclusion Both simultaneous intervention(tDCS and Yunshou)and tDCS followed by Yunshou are more effective on immediate activation of key motor-related cortices in stroke patients with hemiplegia than Yunshou followed by tDCS intervention.

Objective: To understand the incidence risk and risk factors of tuberculosis (TB) among middle school students in Chongqing, so as to provide a basis for formulating TB prevention and control strategies. Methods: From September to December 2022, 32 181 middle school students were selected as the study cohort from 15 administrative districts in Chongqing by using the stratified cluster random sampling method. All cohort members were screened with the tuberculin skin test (TST), and relevant information was collected from January 1, 2023 to December 31, 2024. On the basis of active screening, the follow up data of the participants were compared with the National Tuberculosis Management Information System to obtain the incidence status of the study subjects. The Log rank test was used to compare the TB incidence rates among students with different characteristics, and a Cox proportional hazards model was established to analyze the incidence risk and risk factors of TB. Results: The TST screening rate of the cohort members was 93.0%. During the 2 year follow up period, a total of 36 TB cases occurred, with a cumulative incidence rate of 111.87/100 000 and an incidence density of 55.95/100 000. Among them, the cumulative incidence rate of students from public schools (170.44/ 100 000 ) was higher than that of students from private schools (41.16/100 000), the cumulative incidence rate of students in schools located in high epidemic areas (153.95/100 000) was higher than that in medium epidemic areas (69.00/100 000), and the difference was statistically significant ( χ 2=11.49, 4.73, both P <0.05). The Log-rank test for different TST results showed that the difference in TB comulative incidence rate between students with strongly positive TST results (216.55/ 100 000 ) and those with negative TST results (81.40/100 000) was statistically significant ( χ 2=5.85, P <0.05). Univariate analysis using the Cox proportional hazards model revealed that the risk of TB was lower in students from private schools ( HR=0.25, 95% CI = 0.10-0.59) and students in medium epidemic areas ( HR=0.46, 95%CI =0.23-0.94); whereas the risk of TB was increased in students with strongly positive TST results ( HR=1.39, 95%CI =1.05-1.84) (all P <0.05). Multivariate Cox regression analysis showed that the risk of TB in students from private schools was lower than that of students from public schools ( HR=0.23, 95%CI=0.08-0.62, P <0.05). Conclusions The annual average incidence rate of TB among middle school students in Chongqing is at a relatively high level. It is necessary to strengthen the management and intervention for student groups, including those in public schools, those in schools located in high epidemic areas, and those with strongly positive TST results, so as to reduce the incidence rate of TB.

Objective:To construct a method for hepatitis B virus (HBV) DNA detection based on recombinase-mediated isothermal amplification (RAA)-clustered regularly interspaced short palindromic repeats and their associated protein 13a (CRISPR-Cas13a).Methods:Through the alignment and screening of HBV DNA sequences, a positive plasmid was constructed, and recombinase-aided amplification (RAA) primers and CRISPR RNA (crRNA) were designed. A method for detecting HBV DNA based on the RAA-CRISPR-Cas13a system was developed, and the specificity and sensitivity were evaluated. Utilizing the CRISPR-Cas13a system, 70 clinical samples from HBV DNA-positive patients with various viral loads collected at Beijing You′an Hospital from 2019 to 2021 were analyzed. The detection results were further compared with those results using real-time quantitative polymerase chain reaction (qPCR).Results:The optimal RAA amplification primers and crRNA were first screened using the RAA-CRISPR-Cas13a method, with the sensitivities for detecting HBV DNA standards and for clinical samples at 1 IU/ml and<10 IU/ml, respectively, demonstrating specificity for HBV DNA detection. Compared with qPCR (the gold standard), the detection consistency between the two methods was 100% (70/70).Conclusion:This study established a method for detecting HBV DNA by integrating recombinase-aided amplification (RAA) technology with CRISPR/Cas13a technology.

Objective:To construct a method for hepatitis B virus (HBV) DNA detection based on recombinase-mediated isothermal amplification (RAA)-clustered regularly interspaced short palindromic repeats and their associated protein 13a (CRISPR-Cas13a).Methods:Through the alignment and screening of HBV DNA sequences, a positive plasmid was constructed, and recombinase-aided amplification (RAA) primers and CRISPR RNA (crRNA) were designed. A method for detecting HBV DNA based on the RAA-CRISPR-Cas13a system was developed, and the specificity and sensitivity were evaluated. Utilizing the CRISPR-Cas13a system, 70 clinical samples from HBV DNA-positive patients with various viral loads collected at Beijing You′an Hospital from 2019 to 2021 were analyzed. The detection results were further compared with those results using real-time quantitative polymerase chain reaction (qPCR).Results:The optimal RAA amplification primers and crRNA were first screened using the RAA-CRISPR-Cas13a method, with the sensitivities for detecting HBV DNA standards and for clinical samples at 1 IU/ml and<10 IU/ml, respectively, demonstrating specificity for HBV DNA detection. Compared with qPCR (the gold standard), the detection consistency between the two methods was 100% (70/70).Conclusion:This study established a method for detecting HBV DNA by integrating recombinase-aided amplification (RAA) technology with CRISPR/Cas13a technology.

Objective To examine the association between lymphocyte subsets and renal clinicopathological characteristics as well as prognosis in patients with IgA nephropathy(IgAN).Methods The retrospective analysis included general clinical data and pathological examination results of IgAN patients diagnosed by renal biopsy at the Fourth Hospital of Hebei Medical University from January 2018 to January 2022.Correlation tests were conducted to examine the relationship between lymphocyte subsets and other significant clinicopathological parameters.The optimal cut-off value of CD4+T determined using the Youden index,and patients were grouped accordingly.Kaplan-Meier survival curves and Cox regression analyses were employed to compare the low and high CD4+T lymphocyte groups among IgAN patients,identifying factors influencing renal function progression.The endpoint event was defined as a decrease in estimated glomerular filtration rate(eGFR)of≥30%from baseline,progression to end-stage renal disease(ESRD)[eGFR<15 mL/(min·1.73 m2)or initiation of renal replacement therapy],or all-cause mortality.Results Low CD4+T lymphocytes were significantly positively correlated with blood IgA levels and the proportion of glomerular crescents in IgAN patients(all P<0.05).This study included a total of 53 IgAN patients,divided into two groups based on CD4+T lymphocyte counts:20 patients in the low CD4+T lymphocyte group and 33 patients in the high CD4+T lymphocyte group.In the low CD4+T lymphocyte group,there was a higher proportion of males and a lower proportion of glomerular crescents(P<0.05).Kaplan-Meier survival analysis revealed that patients with low CD4+lymphocytes had a significantly lower cumulative renal survival rate(Log-Rank test χ2=4.188,P=0.041).Cox regression analysis indicated that low CD4+lymphocytes were an independent risk factor for the progression of renal function decline in IgAN patients(HR=2.614,95%CI:1.006～6.788,P=0.048).Conclusions Patients with higher levels of CD4+T lymphocytes exhibit a lower risk of adverse renal outcomes.In contrast,patients with IgA nephropathy and low CD4+T lymphocyte counts tend to have poorer renal survival rates.