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.

Background: Sex-based differences often influence the therapeutic efficacy and safety of medications. Semen Cuscutae is a traditional tonic botanical drug with sex-specific characteristics, traditionally indicated for conditions such as impotence (exclusive to males) and restless fetus (exclusive to pregnant females). However, most existing studies have focused on a single sex. Objective: To evaluate the sex-specific biological effects of Semen Cuscutae in rats and explore its molecular mechanisms, with the aim of uncovering its pharmacological characteristics through a multiomics approach. Methods: A traditional aqueous extract of Semen Cuscutae (SCA) was used as the experimental material. Forty adult Sprague-Dawley rats (equal numbers of males and females) were randomly divided into 4 groups: male control, male SCA treatment (240 mg/kg), female control, and female SCA treatment (240 mg/kg), with 10 rats in each group. The biological effects were comprehensively evaluated using a combination of open field test, biochemical analyses, proteomics, and gut microbiota profiling. Results: As a tonic botanical drug, SCA appeared to directly affect the mental and behavioral state of rats. It significantly altered the time spent by rats in the center area during the open field test, showing a sex-dependent reversal of behaviors. Proteomic analysis of brain tissue identified 624 differentially expressed proteins across the groups, with 10 key differentially expressed proteins related to sex differences, including fibroblast growth factor receptor 3, transcription elongation factor A protein-like 1, 40S ribosomal protein S25, neural cell adhesion molecule, and anion exchange protein 2 (SLC4A2). Enrichment analysis revealed that in male rats, SCA upregulated proteins involved in biological processes such as ribosome function and energy derivation, supporting protein synthesis and enhancing energy supply, showing an overall gain effect. In contrast, in female rats, SCA downregulated proteins associated with processes such as positive regulation of target of rapamycin (TOR) signaling and vesicle transport, suggesting suppression of neuronal signaling and material transport, indicative of a shift toward a more restrained physiological state. Furthermore, SCA reduced gut microbiota diversity in female rats but increased it in males, including the abundance of Akkermansia, which may serve as a crucial mediator. Conclusion: Overall, the biological effects of SCA differ significantly between male and female rats, with evidence suggesting greater health benefits in males. These findings help elucidate the scientific basis of its traditional applications and provide guidance for the precise application of SCA as a functional health food.

Liver fibrosis is the common pathological process in the progression of various chronic liver diseases to liver cirrhosis， and it greatly affects the prognosis of patients with chronic liver diseases. As a novel adipokine， Chemerin participates in the metabolism of glucose and lipids and inflammation， and various studies have shown that the expression level of Chemerin is correlated with the degree of liver fibrosis， suggesting that Chemerin may be involved in the process of liver fibrosis by regulating metabolism and inflammation. Chemerin has shown certain potential in the auxiliary diagnosis of liver fibrosis and the intervention against the progression of liver fibrosis. This article reviews the potential role and mechanism of action of Chemerin in the process of liver fibrosis， in order to provide new ideas for the diagnosis and treatment of liver fibrosis.

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.

Acute pancreatitis (AP) is a life-threatening gastrointestinal disorder for which no effective pharmacological treatments are currently available. One of the pharmacological targets that merits further research is the neurokinin 1 receptor (NK1R), which is found on pancreatic acinar cells and responds to the neuropeptide substance P (SP) that participates in AP. Although a few studies have stated the involvement of SP/NK1R in neurogenic inflammation in AP development, the regulatory mechanism remains unclear. In this study, we found that following activation of NK1R by SP, β-arrestin1, a scaffold protein of NK1R, down-regulated transcription of Adss, Adsl, and Ampd in the purine nucleotide cycle, thereby inhibiting mitochondrial function through fumarate depletion. Interestingly, we identified magnolol as a new and natural NK1R inhibitor with a non-nitrogenous biphenyl core structure. It exhibited a beneficial effect on AP by restoring purine nucleotide cycle metabolic enzymes and fumarate levels. Our study not only provides new therapeutic strategies, leading compounds, and drug translation possibilities for AP, but also provides important clues for the study of downstream mechanisms driven by SP in other diseases.

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 explore the design and implementation effectiveness of the course of Basic Geriatric Care Skills for undergraduate nursing students.Methods:The 496 undergraduate nursing students enrolled in 2019 at The First Affiliated Hospital of Chongqing Medical University were selected as the observation group, while 668 undergraduate nursing students enrolled in 2018 served as the control group. The control group received traditional teaching method, whereas the observation group was additionally provided with the course of Basic Geriatric Care Skills. The course was designed following the five steps of the ADDIE (analysis, design, development, implement, evaluation) model. The design was evaluated using the Delphi expert consultation method. Evaluation of teaching effectiveness included care skill assessment and performance evaluation. The overall evaluation of the course by students was collected through a questionnaire on the Superstar platform. SPSS 25.0 was used for χ2 test and independent samples t test. Results:The course consisted of 8 credit hours and was delivered through both online and offline approaches. The course included 4 modules with 20 care skills. The Cronbach's α values for the two rounds of expert consultation were 0.972 and 0.873, and the Kendall's W values were 0.124 ( χ2=61.38, P<0.05) and 0.260 ( χ2=128.83, P<0.001), respectively. The mean score of care skill assessment was 95.65±2.99 in the observation group and 94.16±3.52 in the control group, and the difference was statistically significant ( P<0.001). In performance evaluation, the highest-scoring dimension was teamwork (19.18±0.88), while the lowest was practical application (18.51±0.94). Questionnaire results indicated that 97.31% (289) of students found the course helpful in learning geriatric care, and 85.86% (255) of students deemed the course necessary. Conclusions:The course of Basic Geriatric Care Skills was well designed to meet the needs of students and to help improve their geriatric care skills.

Objective:To explore the design and implementation effectiveness of the course of Basic Geriatric Care Skills for undergraduate nursing students.Methods:The 496 undergraduate nursing students enrolled in 2019 at The First Affiliated Hospital of Chongqing Medical University were selected as the observation group, while 668 undergraduate nursing students enrolled in 2018 served as the control group. The control group received traditional teaching method, whereas the observation group was additionally provided with the course of Basic Geriatric Care Skills. The course was designed following the five steps of the ADDIE (analysis, design, development, implement, evaluation) model. The design was evaluated using the Delphi expert consultation method. Evaluation of teaching effectiveness included care skill assessment and performance evaluation. The overall evaluation of the course by students was collected through a questionnaire on the Superstar platform. SPSS 25.0 was used for χ2 test and independent samples t test. Results:The course consisted of 8 credit hours and was delivered through both online and offline approaches. The course included 4 modules with 20 care skills. The Cronbach's α values for the two rounds of expert consultation were 0.972 and 0.873, and the Kendall's W values were 0.124 ( χ2=61.38, P<0.05) and 0.260 ( χ2=128.83, P<0.001), respectively. The mean score of care skill assessment was 95.65±2.99 in the observation group and 94.16±3.52 in the control group, and the difference was statistically significant ( P<0.001). In performance evaluation, the highest-scoring dimension was teamwork (19.18±0.88), while the lowest was practical application (18.51±0.94). Questionnaire results indicated that 97.31% (289) of students found the course helpful in learning geriatric care, and 85.86% (255) of students deemed the course necessary. Conclusions:The course of Basic Geriatric Care Skills was well designed to meet the needs of students and to help improve their geriatric care skills.

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 establish a rapid and accurate method for the detection of Klebsiella pneumoniae carbapenemase (KPC) carbapenemase gene based on recombinase aided amplification (RAA)-CRISPR-Cas13a (CRISPR-Cas13a) technology. Methods:Twenty-five clinical isolates of carbapenem-resistant Klebsiella pneumoniae (CRKP) and five carbapenem-sensitive Klebsiella pneumoniae (CSKP) strains preserved in 2020-2021 in Beijing Chuiyangliu Hospital were randomly collected, and the total DNA samples of the strains was extracted. RAA primers specific for KPC DNA and CRISPR RNA (crRNA) were designed to establish a rapid and accurate method for the detection of KPC carbapenemase gene based on RAA-CRISPR-Cas13a technology. The method was evaluated by plasmids and clinical sample strains, and the detection was also performed by Quantitative real-time PCR (qPCR) method to compare the detection rate and consistency of the two methods. Results:The RAA-CRISPR-Cas13a method can detect KPC plasmids and samples with a sensitivity of 1 copy/μl, which is higher than that of qPCR (10 1 copies/μl). Among the 30 clinical strains (including 25 CRKP strains and 5 CSKP strains), 23 strains were detected to carry KPC gene by both RAA-CRISPR-Cas13a method and qPCR method, and 7 strains were not detected with KPC gene. The detection rate of KPC gene in the 25 CRKP strains was 92% (23/25). The positive coincidence rate of the two methods was 100% (23/23). Conclusions:This study combined RAA amplification technology with CRISPR-Cas13a technology to establish a rapid and accurate method for detecting KPC carbapenemase gene. The method is useful for accurate screening of KPC carbapenemase-producing strains. It has a wide application prospect in drug resistance monitoring and infection control.