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.

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 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.

Objective:To know the current situation of kinesiophobia in patients after cardiac surgery under cardiopulmonary bypass, and to clarify its influencing factors, so as to provide reference for developing intervention strategies to improve kinesiophobia level.Methods:This was a cross-sectional study. From February 2022 to September 2022, the patients after cardiac valve surgery under cardiopulmonary bypass in the Second Affiliated Hospital Zhejiang University School of Medicine were investigated by convenience sampling methods. The survey was conducted using the General Information Questionnaire, The Tampa Scale for Kinesiophobia Heart, Exercise Self-Efficacy Scale, Pain Catastrophizing Scale, and Adaptation, Partnership, Growth, Affection and Resolve(APGAR) as research tools, and the influencing factors were analyzed using univariate and binary Logistic regression analysis.Results:A total of 219 patients were included, of which 97 patients (44.3%) had kinesiophobia. The results of binary Logistic regression analysis showed that monthly family income level, first time out of bed after operation, fear of falling, the family APGAR, and pain catastrophizing were significant influencing factors of kinesiophobia in patients after heart valve surgery under cardiopulmonary bypass (all P<0.05). Conclusions:The prevalence of kinesiophobia is high among patients after heart valve surgery under cardiopulmonary bypass. Clinicians should pay attention to patients with low monthly family income level, late first time out of bed after surgery, and fear of falling, as well as strengthen communication with patients and families, focus on the management of acute postoperative pain. In order to reduce or avoid the occurrence of kinesiophobia and enable patients to benefit from early ambulation.

Objective:To investigate the application effect of virtual simulation combined with flipped classroom in experimental teaching of the nursing care of falls in the elderly.Methods:The 497 nursing undergraduates in the class of 2019 in Chongqing Medical University were selected as subjects and were then divided into control group (two classes with 251 students) and intervention group (two classes with 246 students). The students in the control group received traditional experimental teaching, and those in the intervention group received blended experimental teaching with virtual simulation combined with flipped classroom. At the end of the course, the use of virtual simulation platform was analyzed for the students in the intervention group, and a questionnaire survey and theoretical examination were used to compare the effect of experimental teaching between the two groups. SPSS 27.0 was used for the t-test and the chi-square test. Results:The frequency of use of the virtual simulation platform was (2.65±1.38) times per person in the intervention group, with an online learning time of 54.12-147.32 minutes. The questionnaire survey showed that compared with the control group, the intervention group had significantly higher scores of the achievement of teaching objectives and teaching satisfaction ( P<0.05). Compared with the control group after teaching, the intervention group had significantly higher scores of teaching promotion in terms of stimulating learning interest, cultivating self-learning ability, developing clinical thinking ability, improving innovation, and enhancing health education ability ( P<0.05). The intervention group had a significantly higher theoretical examination score than the control group [(79.38±5.09) vs. (77.88±4.97), P<0.05]. Conclusions:In the blended experimental teaching of the nursing care of falls in the elderly, virtual simulation combined with flipped classroom can help students master related knowledge and skills and cultivate their self-learning ability, clinical thinking ability, innovation ability, and health education ability.

【Objective】 To investigate the risk factors of vasovagal reactions(VVR) related to plasma donation, so as to put forward clinical suggestions for early identification and accurate intervention of high-risk groups to ensure the safety of plasma donation. 【Methods】 The demographic characteristics(i.e. gender, age, weight) and records of plasma donors(donation history, pulse before plasma donation, duration of collection, etc.) were collected from July to December 2019 in a region of Sichuan. Based on logistic regression analysis, the correlation between these factors and the risk of VVR was explored. 【Results】 The information of 69 172 donors was collected, and the incidence of VVR was 7.04‰. The risk of VVR was reduced by 99% in the group with plasma collection duration less than 30 minutes compared with the group with plasma collection duration more than 50 minutes(OR, 0.01; 95% CI, 0.00~0.01; P<0.001). The risk of male group was 94 % lower than that of female group(OR, 0.06; 95% CI, 0.04~0.10; P<0.001). Compared with the 45~50 kg group, the risk of weight greater than 80 kg group decreased by 80%(OR, 0.20; 95% CI, 0.09~0.42; P<0.001). The risk of repeated donation group was 34 % lower than that of the first time donation group(OR, 0.66; 95% CI, 0.47~0.91; P<0.001). The risk of VVR in the group with pulse greater than 90 bpm before plasma donation was 2.43 times that in the 60~69 bmp group(OR, 2.43; 95% CI, 1.75~3.36; P<0.001). 【Conclusion】 Duration of plasma collection, gender, weight, frequency of plasma donation, pulse before plasma donation and donor status are independent risk factors for plasma donation-related VVR. Among them, plasma collection duration, gender and weight were the main independent risk factors for plasma donation-related VVR. For donors with plasma collection duration more than 50 minutes, female and low weight, higher risk of VVR was presented and more preventive intervention should be given.

Objective:To investigate the expression and role of asparte-specific cysteine protease (Caspase)-1, inflammasomes key molecule, in hepatitis B virus (HBV)-related diseases.Methods:HBV-related liver disease patients' serum (438 cases) and liver tissue (82 cases) samples were collected from Beijing You'an Hospital affiliated with Capital Medical University. The mRNA expression level of caspase-1 in liver tissue was detected by real-time fluorescence quantitative PCR (qRT-PCR). The protein expression level of Caspase-1 in liver tissue was detected by the immunofluorescence method. The activity of Caspase-1 was detected using the Caspase-1 colorimetric assay kit. The level of Caspase-1 in the serum was detected by an ELISA kit.Results:The results of qRT-PCR showed that the mRNA level of Caspase-1 was downregulated in patients with chronic hepatitis B (CHB), cirrhosis (LC), and hepatocellular carcinoma (HCC), while up-regulated in patients with acute-on-chronic liver failure (ACLF) ( P＜0.01) compared with normal subjects. Immunofluorescence assays showed that Caspase-1 protein levels were elevated in ACLF patients, decreased in HCC and LC patients, and slightly elevated in CHB patients. The activity of Caspase-1 was slightly higher in liver tissue from CHB, LC, and HCC patients than in the normal control group, and there was no statistically significant difference between the groups. Additionally, compared with the control group, Caspase-1 activity was significantly reduced in the ACLF group ( P＜0.01). Serum Caspase-1 levels were significantly lower in patients with CHB, ACLF, LC, and HCC than in normal subjects, and serum Caspase-1 levels were lowest in patients with ACLF ( P＜0.001). Conclusion:Caspase-1, a key molecule of inflammasomes, plays an important role in HBV-related diseases and has significant differences, showing distinct features for ACLF than other HBV-related diseases.

The imbalance of lipid metabolism is an important factor causing a series of metabolic diseases such as non-alcoholic fatty liver disease (NAFLD) and hyperlipidemia.Liver X receptors is a member of the nuclear receptor superfamily, which maintains cholesterol homeostasis by regulating cholesterol absorption, transport, reverse cholesterol transport, biosynthesis and other functions.It plays an important role in maintaining lipid homeostasis by regulating de novo fat synthesis to maintain the balance of fatty acids in the body.