1.Value of different noninvasive diagnostic models in the diagnosis of esophageal and gastric varices with significant portal hypertension in compensated hepatitis B cirrhosis
Cheng LIU ; Jiayi ZENG ; Mengbing FANG ; Zhiheng CHEN ; Bei GUI ; Fengming ZHAO ; Jingkai YUAN ; Chaozhen ZHANG ; Meijie SHI ; Yubao XIE ; Xiaoling CHI ; Huanming XIAO
Journal of Clinical Hepatology 2025;41(2):263-268
ObjectiveTo investigate the value of different noninvasive diagnostic models in the diagnosis of esophageal and gastric varices since there is a high risk of esophageal and gastric varices in patients with compensated hepatitis B cirrhosis and significant portal hypertension, and to provide a basis for the early diagnosis of esophageal and gastric varices. MethodsA total of 108 patients with significant portal hypertension due to compensated hepatitis B cirrhosis who attended Guangdong Provincial Hospital of Traditional Chinese Medicine from November 2017 to November 2023 were enrolled, and according to the presence or absence of esophageal and gastric varices under gastroscopy, they were divided into esophageal and gastric varices group (GOV group) and non-esophageal and gastric varices group (NGOV group). Related data were collected, including age, sex, imaging findings, and laboratory markers. The chi-square test was used for comparison of categorical data between groups; the least significant difference t-test was used for comparison of normally distributed continuous data between groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups. The receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic value of five scoring models, i.e., fibrosis-4 (FIB-4), LOK index, LPRI, aspartate aminotransferase-to-platelet ratio index (APRI), and aspartate aminotransferase/alanine aminotransferase ratio (AAR). The binary logistic regression method was used to establish a combined model, and the area under the ROC curve (AUC) was compared between the combined model and each scoring model used alone. The Delong test was used to compare the AUC value between any two noninvasive diagnostic models. ResultsThere were 55 patients in the GOV group and 53 patients in the NGOV group. Compared with the NGOV group, the GOV group had a significantly higher age (52.64±1.44 years vs 47.96±1.68 years, t=0.453, P<0.05) and significantly lower levels of alanine aminotransferase [42.00 (24.00 — 17.00) U/L vs 82.00 (46.00 — 271.00) U/L, Z=-3.065, P<0.05], aspartate aminotransferase [44.00 (32.00 — 96.00) U/L vs 62.00 (42.50 — 154.50) U/L,Z=-2.351, P<0.05], and platelet count [100.00 (69.00 — 120.00)×109/L vs 119.00 (108.50 — 140.50)×109/L, Z=-3.667, P<0.05]. The ROC curve analysis showed that FIB-4, LOK index, LPRI, and AAR used alone had an accuracy of 0.667, 0.681, 0.730, and 0.639, respectively, in the diagnosis of esophageal and gastric varices (all P<0.05), and the positive diagnostic rates of GOV were 69.97%, 65.28%, 67.33%, and 58.86%, respectively, with no significant differences in AUC values (all P>0.05), while APRI used alone had no diagnostic value (P>0.05). A combined model (LAF) was established based on the binary logistic regression analysis and had an AUC of 0.805 and a positive diagnostic rate of GOV of 75.80%, with a significantly higher AUC than FIB-4, LOK index, LPRI, and AAR used alone (Z=-2.773,-2.479,-2.206, and-2.672, all P<0.05). ConclusionFIB-4, LOK index, LPRI, and AAR have a similar diagnostic value for esophageal and gastric varices in patients with compensated hepatitis B cirrhosis and significant portal hypertension, and APRI alone has no diagnostic value. The combined model LAF had the best diagnostic efficacy, which provides a certain reference for clinical promotion and application.
2.YAN Huimin's Experience in Staged Treatment of Pediatric IgA Vasculitis
Tianxiang LIU ; Qian ZHAO ; Jing HAO ; Mengtong WANG ; Jiayi LIU ;
Journal of Traditional Chinese Medicine 2025;66(12):1212-1216
This paper summarizes Professor YAN Huimin's clinical experience in the staged treatment of pediatric IgA vasculitis. It is believed that the pathogenesis in the acute stage is characterized by fire and heat entering the blood and damaging the collaterals, and the treatment should focus on clearing heat and stopping bleeding, using the self-formulated Qingzi Liangxue Formulation (青紫凉血方). In the remission stage, the pathogenesis is qi rebellion leading to blood extravasation. The treatment principle is to regulate qi and stabilize blood, using the self-formulated Heqi Ningxue Formulation (和气宁血方). During the protracted stage, the pathogenesis is latent pathogenic factors due to yin deficiency, and the treatment should aim to nourish the collaterals and strengthen the root, with the self-formulated Yangluo Guben Formulation (养络固本方). Meanwhile, the method of promoting blood stasis resolution is consistently applied throughout the entire treatment process.
3.Advances in immunotherapy for biliary tract cancers
Yuhao ZHAO ; Mao YANG ; Jiayi FENG ; Xu’an WANG ; Yingbin LIU
Chinese Medical Journal 2024;137(5):524-532
Biliary tract cancers (BTC), a heterogeneous disease with poor prognosis, including gallbladder cancer (GBC), intrahepatic cholangiocarcinoma (ICC), and extrahepatic cholangiocarcinoma (ECC). Although surgery is currently the primary regimen to treat BTC, most BTC patients are diagnosed at an advanced stage and miss the opportunity of surgical eradication. As a result, non-surgical therapy serves as the main intervention for advanced BTC. In recent years, immunotherapy has emerged as one of the most promising therapies in a number of solid cancers, and it includes immune checkpoint inhibitors (ICIs) monotherapy or combined therapy, tumor vaccines, oncolytic virus immunotherapy, adoptive cell therapy (ACT), and cytokine therapy. However, these therapies have been practiced in limited clinical settings in patients with BTC. In this review, we focus on the discussion of latest advances of immunotherapy in BTC and update the progress of multiple current clinical trials with different immunotherapies.
4.Analysis of the factors influencing prognosis of the adult in-hospital cardiac arrest
Jiayi ZHAO ; Dehua ZENG ; Aiqun ZHU
Chinese Critical Care Medicine 2024;36(4):398-403
Objective:To explore the factors influencing prognosis of patients with in-hospital cardiac arrest (IHCA).Methods:A retrospective observational study was conducted. The clinical data of patients who developed IHCA and underwent cardiopulmonary resuscitation (CPR) at the Second Xiangya Hospital of Central South University from January 1, 2016, to December 31, 2022 were analyzed. The patients' information, including gender, age, medical history, pre-cardiac arrest related parameters [1-hour pre-cardiac arrest neurological function, 24-hour pre-cardiac arrest hemoglobin (Hb) levels, 1-hour pre-cardiac arrest vital signs], initial CPR-related factors (implementation time and location, initial rhythm, ventilation method, defibrillation and resuscitation drugs) as well as restoration of spontaneous circulation (ROSC) related parameters (vital signs at ROSC and 1 hour after ROSC, 24-hour post-cardiac arrest Hb, and IHCA events), were collected through the hospital's electronic medical record system. The clinical data were compared between ROSC and non-ROSC patients as well as between patients with favorable neurological function [cerebral performance category (CPC) grades 1-2] and unfavorable neurological function (CPC grades 3-5) at 28 days. The factors with statistical significance in univariate analysis and clinical significance were enrolled in a binary multivariate Logistic regression model to analyze the influencing factors of ROSC and neurological function at 28 days after ROSC. The predictive value of factors influencing neurological function at 28 days was assessed using receiver operator characteristic curve (ROC curve).Results:A total of 277 IHCA-CPR patients were enrolled, of which 230 achieved ROSC (83.0%) and 47 were not achieved (17.0%). Compared with non-ROSC patients, ROSC patients had lower prevalence of cerebrovascular disease history and proportion of adrenaline usage, but a higher proportion of initial shockable rhythms. In the multivariate Logistic regression analysis, it was found that using a bag-mask ventilation+endotracheal intubation (compared with a bag-mask ventilation alone) was beneficial for achieving ROSC in IHCA-CPR patients [odds ratio ( OR) = 2.895, 95% confidence interval (95% CI) was 1.204-6.962, P = 0.018], while a initial non-shockable rhythm was not conducive to achieving ROSC in IHCA-CPR patients ( OR = 0.349, 95% CI was 0.147-0.831, P = 0.017). Among the 230 ROSC patients, 42 had good neurological function at 28 days (18.3%), and 188 had poor neurological function (81.7%). Compared with the patients with good neurological function, the patients with the poor neurological function were older and had a higher prevalence of 1-hour pre-cardiac arrest neurological dysfunction and low perfusion, initial non-shockable rhythms, endotracheal intubation, and usage of adrenaline, vasopressors and sodium bicarbonate, a lower proportion of defibrillation and antiarrhythmic medication usage as well as lower 24-hour post-cardiac arrest Hb levels. The multivariate Logistic regression analysis revealed that female ( OR = 6.449, 95% CI was 1.837-22.642, P = 0.004), older age ( OR = 1.054, 95% CI was 1.017-1.093, P = 0.004), 1-hour pre-cardiac arrest neurological dysfunction ( OR = 25.044, 95% CI was 2.737-229.169, P = 0.004), 1-hour pre-cardiac arrest low perfusion ( OR = 3.880, 95% CI was 1.306-11.524, P = 0.028), endotracheal intubation (compared with a bag-mask ventilation; OR = 8.712, 95% CI was 1.402-54.141, P = 0.020) and face mask+endotracheal intubation during CPR (compared with a bag-mask ventilation; OR = 11.089, 95% CI was 3.482-35.320, P = 0.000), IHCA events > 1 time ( OR = 4.221, 95% CI was 1.249-14.226, P = 0.020) were positively associated with poor neurological function at 28 days, which were independent risk factors those were not conducive to 28-day neurological function recovery after ROSC in IHCA-CPR patients. In contrast, usage of antiarrhythmic medication ( OR = 0.345, 95% CI was 0.134-0.890, P = 0.028) and 24-hour post-cardiac arrest Hb ( OR = 0.983, 95% CI was 0.966-0.999, P = 0.043) were negatively associated with poor neurological function at 28 days, which were protective factors those were beneficial for the recovery of neurological function. ROC curve analysis showed that the area under the ROC curve (AUC) of 24-hour post-cardiac arrest Hb for predicting poor neurological function at 28 days after ROSC in IHCA-CPR patients was 0.659 (95% CI was 0.577-0.742), with a cut-off value of 99.5 g/L (sensitivity was 76.2%, specificity was 57.8%). Conclusions:Defibrillation and tracheal intubation during CPR are crucial for IHCA patients. It was also observed that patients with low Hb (< 99.5 g/L should be of high concern), older age, 1-hour pre-cardiac arrest neurological function and hypoperfusion, and IHCA events > 1 time were significantly related to unfavorable neurological outcome in adult resuscitated patients with IHCA.
5.Application value of antegrade splenic superior region dissection first in laparoscopic total gastrectomy of obesity gastric cancer
Danhua XU ; Jiayi GU ; Xinli MA ; Chunchao ZHU ; Ming WANG ; Enhao ZHAO ; Zizhen ZHANG ; Jiangfeng QIU ; Hui CAO
Chinese Journal of Digestive Surgery 2024;23(4):609-612
Objective:To investigate the application value of antegrade splenic superior region dissection first in laparoscopic total gastrectomy of obesity gastric cancer.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 21 obesity patients with gastric cancer who underwent laparoscopic total gastrectomy in Renji Hospital of Shanghai Jiaotong University School of Medicine from July 2018 to October 2023 were collected. There were 16 males and 5 females, aged (58±13)years. All 21 patients underwent laparoscopic total gastrec-tomy with antegrade splenic superior region dissection first. Observation indicators: operation time, volume of intraoperative blood loss, laparotomy conversion, intraoperative splenic hemorrhage or gastric hemorrhage, lymph node dissection, time to postoperative first flatus, time to postoperative initial liquid food intake, duration of postoperative hospital stay, postoperative complication. Measure-ment data with normal distribution were represented as Mean± SD, and count data were expressed as absolute numbers. Results:All 21 patients underwent laparoscopic total gastrectomy success-fully, with the operation time of (283±47)minutes, time for splenogastric ligament and vascular manage-ment of (34±12)minutes, volume of intraoperative blood loss of (143±86)mL, and no laparotomy conversion. There was no intraoperative splenic hemorrhage or gastric haemorrhage. The total number of lymph node dissected in 21 patients was 375, with the number of lymph node dissected as (21±9)per case. Time to postoperative first flatus, time to postoperative initial liquid food intake and duration of postoperative hospital stay in 21 patients were (3.1±0.7)days, (4.0±0.8)days and (10.1±3.0)days, respectively. There were 2 patients with postoperative complications, including 1 case of incision infection and 1 case of lung infection. The 2 patients with postoperative com-plications were recovered and discharged after conservative treatment. There was no death during the postoperative 30 days.Conclusion:The application of antegrade splenic superior region dissec-tion first in laparoscopic total gastrectomy is safe and feasible, which can reduce surgical difficulty.
6.Pueraria Lobata Flowers Extract inhibits THP-1-derived foam cell formation by acti-vating PPARγ to upregulate the expression of ABCA1
Rongrong ZHU ; Mengjiao CHEN ; Zhenwang ZHAO ; Jiayi LIU ; Jianfeng WU ; Yufei WANG ; Min ZHANG
Chinese Journal of Arteriosclerosis 2024;32(5):395-401
Aim To explore the effect of Pueraria Lobata Flowers Extract(PFE)on lipid accumulation in mac-rophage-derived foam cells.Methods The concentration of PFE in THP-1-derived foam cells was screened by MTT,intracellular lipid accumulation was detected by oil red O staining and cholesterol detection kit,intracellular cholesterol ef-flux levels were detected by cholesterol efflux assay kit,RT-qPCR and Western blot were used to analyze mRNA and pro-tein expression.Results PFE significantly reduced lipid accumulation in THP-1-derived foam cells.PFE did not affect the mRNA expression of CD36,scavenger receptor-A Ⅰ(SR-A Ⅰ),sterol regulatory element-binding protein 2(SREBP2),3-hydroxy-3-methylglutaryl-CoA reductase(HMGCR),but it could upregulate the mRNA and protein expres-sion levels of ATP-binding cassette transporter A1(ABCA1)(P<0.05),and promote the intracellular cholesterol efflux of macrophage-derived foam cells(P<0.01).PFE could activate the activity of peroxisome proliferator-activated receptor y(PPARγ)(P<0.01)and upregulate the mRNA and protein expression levels of PPARγ(P<0.05).Compared with the PFE control group,the expression of PPARγ and ABCA1 proteins decreased and cholesterol efflux decreased after GW9662 treatment(all P<0.01).Conclusion PFE could significantly prevent the lipid accumulation in THP-1-derived foam cells and inhibit the formation of foam cells by upregulating ABCA1 expression and cholesterol efflux mediated by PPARγ.
7.Impact of bladder volume on dosimetry of CTV and OAR in localized prostate cancer treated with proton therapy
Danni WANG ; Huan LI ; Cheng XU ; Wendong FAN ; Mei CHEN ; Xiaofang QIAN ; Dawei QIN ; Chensheng SHI ; Ruozhui ZHAO ; Weixiang QI ; Qiyun HUANG ; Jiayi CHEN ; Lu CAO
Chinese Journal of Radiation Oncology 2024;33(6):524-531
Objective:To evaluate the impact of bladder volume on dosimetric parameters of clinical target volume (CTV) and organs at risk (OAR) of intensity modulated proton therapy (IMPT) for localized prostate cancer during the treatment planning and daily treatment.Methods:Clinical data of 25 patients with localized prostate cancer admitted to Ruijin Hospital affiliated to Shanghai Jiao Tong University School of Medicine from November 2021 to June 2022 and enrolled in the "Proton Therapy System" (SAPT-PS-01) registered clinical trial were retrospectively analyzed. All patients were male and the median age was 72 years old. A total of 30 sets of IMPT plans were obtained. Based on the planning CT (30 sets) and weekly verification CT during treatment (172 sets), bladder volume, CTV and OAR dose parameters were collected. Spearman correlation analysis was used to evaluate the correlation between bladder volume in CT and the dosimetric parameters of CTV and OAR during IMPT plans, and Wilcoxon-Mann-Whitney test was adopted to compare the dosimetric parameters of CTV and OAR among different bladder volume change groups.Results:The V 95% of CTV1 and CTV2 were both 100.0%±0.0% in IMPT plans. Bladder volume was significantly negatively correlated with D mean, V 70 Gy(RBE), V 60 Gy(RBE), V 50 Gy(RBE), V 40 Gy(RBE) of the bladder ( P<0.001, 0.003, <0.001, <0.001,<0.001), and D mean, V 50 Gy(RBE) of the small intestine (both P<0.001). During treatment, bladder D mean, V 70 Gy(RBE), V 60 Gy(RBE), V 50 Gy(RBE), V 40 Gy(RBE)( P<0.001, 0.001, <0.001, <0.001, <0.001), rectal D mean, V 50 Gy(RBE), V 40 Gy(RBE) (all P<0.001), small intestine D mean, V 50 Gy (RBE) (both P<0.001) of patients with bladder volume increase >20% compared to baseline were significantly decreased compared to those in IMPT plans. But CTV1 V 100%, and CTV2 V 95% were significantly decreased too( P=0.029, 0.020). In the bladder volume decreased>20% patients, the D mean, V 70 Gy(RBE), V 60 Gy(RBE), V 50 Gy(RBE), V 40 Gy(RBE) of the bladder were significantly increased compared to those in IMPT plans (all P<0.001). However, a bladder volume reduction of ≤20% and increase of ≤20% from baseline had no significant impact on CTV and OAR dosimetric parameters during treatment. Conclusions:For patients with localized prostate cancer undergoing proton therapy, a certain bladder volume should be ensured during planning CT scans. During the daily treatment, the bladder volume should be maintained between 80%-120% of the baseline level to ensure CTV coverage and good dose sparing to OAR.
8.Stellate Ganglion Block as an Adjunctive Intervention for Chronic Subjective Tinnitus: Efficacy and Predictive Indicators
Zhicheng LI ; Nan CHENG ; Jibin XING ; Jiawang TIAN ; Jianqi ZHAO ; Huajing TIAN ; Jiayi LIN ; Xiangli ZENG
Journal of Sun Yat-sen University(Medical Sciences) 2024;45(2):276-282
ObjectiveTo explore the efficacy and predictive indicators of stellate ganglion block (SGB) as an adjunctive intervention for chronic subjective tinnitus and accumulate experience for the application of SGB in the clinical treatment of tinnitus. MethodsA retrospective review was conducted on the data of chronic subjective tinnitus patients who received SGB intervention, with unsatisfactory outcomes otherwise. Pure tone audiometry (PTA), tinnitus loudness evaluation and Pittsburgh sleep quality index (PSQI) were used. The tinnitus handicap inventory (THI) scores were compared before and after SGB intervention. Correlation analysis and linear regression equations were employed to identify the potential indicators predicting the effectiveness of SGB intervention. Statistical analysis was performed by SPSS 24.0 software. ResultsBy April 2023, a total of 107 patients with chronic subjective tinnitus had undergone SGB intervention, including 67 male and 40 female, with a mean age of (45.32±11.40) years old and an average tinnitus history of (20.32±24.64) months [16 (12~20)]. Only 7 patients (6.54%) quitted the intervention for personal reasons, which demonstrated good compliance with the intervention. No patients experienced adverse reactions such as infection at the injection site, hematoma, nerve injury, local anesthetic intoxication and so on, which revealed good safety. After SGB intervention, THI scores decreased to below 36 points in 77 patients and decrease by 10 points or more in 12 of the remaining patients, with a total effective rate of 89%. A paired sample t-test showed a significant difference in THI scores before and after SGB intervention (t=15.575, P<0.001), indicating good improvement. Pearson correlation analysis suggested that pre-intervention THI scores and subjective tinnitus loudness were significantly positively correlated with the improvement level of THI scores (P<0.05). Further stepwise linear regression analysis found that "pre-intervention THI scores" had statistical significance (P<0.001), with a regression coefficient of 0.308, predicting a 17.4% improvement level in THI scores. ConclusionsDue to its good and safe short-term effects, SGB intervention can be used as a supplementary option for chronic subjective tinnitus when other interventions are not ideal, especially for patients with higher THI scores. However, further research is needed to clarify the long-term efficacy and underlying mechanisms, in order to establish a more solid theoretical basis for SGB intervention in the treatment of subjective tinnitus.
9.Modified Xiaoyaosan Alleviates Depression-like Behaviors by Regulating Activation of Hippocampal Microglia Cells in Rat Model of Juvenile Depression
Jiayi SHI ; Yun XIANG ; Ziyang ZHOU ; Dahua WU ; Feng QIU ; Chang LEI ; Hongyu ZENG ; Kaimei TAN ; Hongqing ZHAO ; Dong YANG ; Yuhong WANG ; Pengxiao GUO ; Xiuli ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):46-56
ObjectiveTo investigate the mechanism of Baihuan Xiaoyao Decoction (Xiaoyaosan added with Lilii Bulbus and Albiziae Cortex) in alleviating depression-like behaviors of juvenile rats by regulating the polarization of microglia. MethodSixty juvenile SD rats were randomized into normal control, model, fluoxetine, and low-, medium-, and high-dose (5.36, 10.71, 21.42 g·kg-1, respectively) Baihuan Xiaoyao decoction groups. The rat model of juvenile depression was established by chronic unpredictable mild stress (CUMS). The sucrose preference test (SPT) was carried out to examine the sucrose preference of rats. Forced swimming test (FST) was carried out to measure the immobility time of rats. The open field test (OFT) was conducted to measure the total distance, the central distance, the number of horizontal crossings, and the frequency of rearing. Morris water maze (MWM) was used to measure the escape latency and the number of crossing the platform. The immunofluorescence assay was employed to detect the expression of inducible nitric oxide synthase (iNOS, the polarization marker of M1 microglia) and CD206 (the polarization marker of M2 microglia). Real-time polymerase chain reaction was employed to determine the mRNA levels of iNOS, CD206, pro-inflammatory cytokines [tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6] and anti-inflammatory cytokines (IL-4 and IL-10) in the hippocampus. Western blotting was employed to determine the protein levels of iNOS and CD206 in the hippocampus. The levels of IL-4 and IL-6 in the hippocampus were detected by enzyme-linked immunosorbent assay. ResultCompared with the normal control group, the model rats showed a reduction in sucrose preference (P<0.05), an increase in immobility time (P<0.05), decreased motor and exploratory behaviors (P<0.05), and weakened learning and spatial memory (P<0.05). In addition, the model rats showed up-regulated mRNA and protein levels of iNOS and mRNA levels of IL-1β, IL-6, and TNF-α (P<0.05). Compared with the model group, Baihuan Xiaoyao decoction increased the sucrose preference value (P<0.05), shortened the immobility time (P<0.01), increased the motor and exploratory behaviors (P<0.05), and improved the learning and spatial memory (P<0.01). Furthermore, the decoction down-regulated the positive expression and protein level of iNOS, lowered the levels of TNF-α, IL-1β, and IL-6 (P<0.01), promoted the positive expression of CD206, and elevated the levels of IL-4 and IL-10 (P<0.01) in the hippocampus of the high dose group. Moreover, the high-dose Baihuan Xiaoyao decoction group had higher sucrose preference value (P<0.01), shorter immobility time (P<0.01), longer central distance (P<0.01), stronger learning and spatial memory (P<0.01), higher positive expression and protein level of iNOS (P<0.01), lower levels of TNF-α, IL-1β, and IL-6 (P<0.05, P<0.01), lower positive expression and mRNA level of iNOS (P<0.05), and higher levels of IL-4 and IL-10 (P<0.05, P<0.01) than the fluoxetine group. ConclusionBaihuan Xiaoyao decoction can improve the depression-like behavior of juvenile rats by inhibiting the M1 polarization and promoting the M2 polarization of microglia in the hippocampus.
10.Menaquinone-4 exerts a protective effect against carbon tetrachloride-induced acute liver injury in mice by alleviating ferroptosis
Lu YE ; Fan ZHAO ; Qianqian HUANG ; Jiayi ZHANG ; Jianqing WANG
Journal of Clinical Hepatology 2024;40(1):121-128
ObjectiveTo investigate whether menaquinone-4 (MK-4) can exert a protective effect against carbon tetrachloride (CCl4)-induced acute liver injury (ALI) in mice by alleviating ferroptosis. MethodsAfter adaptive feeding, adult male ICR mice, aged 8 weeks, were divided into Control group, MK-4 group, CCl4 model group (6-hour, 12-hour, and 24-hour), and MK-4+CCl4 group (6-hour, 12-hour, and 24-hour), with 6 mice in each group. The mice in the Control group were given intraperitoneal injection of an equal dose of corn oil; the mice in the MK-4 group were given intraperitoneal injection of 40 mg/kg MK-4 solution, followed by an equal dose of corn oil after 1 hour; the mice in the MK-4+CCl4 group (6-hour, 12-hour, and 24-hour) were given intraperitoneal injection of 40 mg/kg MK-4 solution, and after 1 hour, the mice in this group and the CCl4 model group (6-hour, 12-hour, and 24-hour) were given intraperitoneal injection of 0.3 mL/kg CCl4 solution, with samples collected at 6, 12, and 24 hours. HE staining was used to observe the pathological changes of mouse liver; Prussian blue staining was used to observe iron accumulation in liver tissue; a biochemical analyzer was used to measure the serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT); related kits were used to measure the levels of tissue iron content and the oxidative stress indices malondialdehyde (MDA) and glutathione (GSH) in liver homogenate; RT-PCR was used to measure the expression levels of ferroptosis marker genes (acyl-CoA synthetase long-chain family member 4 [ACSL4], prostaglandin-endoperoxide synthase 2 [PTGS2], and glutathione peroxidase 4 [GPX4]) and iron metabolism-related genes (hemojuvelin [HJV], transferrin receptor 1 [TFR1], and ferroportin [FPN]), and Western blot was used to measure the protein expression level of GPX4. 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. ResultsIn the aging study, compared with the Control group, the CCl4 model group (6-hour, 12-hour, and 24-hour) had significant increases in liver weight coefficient and the serum levels of ALT and AST (all P<0.05), and HE staining also showed that liver injury gradually aggravated over time. Meanwhile, compared with the CCl4 model group (6-hour, 12-hour, and 24-hour), the MK-4+CCl4 (12-hour) group had significant reductions in liver weight coefficient and the serum levels of ALT and AST (all P<0.05), with a reduction in the necrotic area of liver tissue, and therefore, 12-hour mouse tissue samples were used for detection in the following study. Compared with the Control group, the CCl4 group had a significant increase in MDA and a significant reduction in GSH (both P<0.05), and compared with the CCl4 group, the MK-4+CCl4 group had a significant reduction in MDA and a significant increase in GSH (both P<0.05). Compared with the Control group, the CCl4 group had significant increases in the key ferroptosis indices ASCL4 and PTGS2 and a significant reduction in GPX4 (all P<0.05); compared with the CCl4 group, the MK-4+CCl4 group had significant reductions in the mRNA expression levels of ASCL4 and PTGS2 and a significant increase in the mRNA expression level of GPX4 (all P<0.05). Western blotting showed that compared with the Control group, the CCl4 group had a significant reduction in the protein expression level of GPX4 (P<0.05), and compared with the CCl4 group, the MK-4+CCl4 group had a significant increase in the protein expression level of GPX4 (P<0.05). Prussian blue staining showed that compared with the Control group, the CCl4 group had a significant increase in iron accumulation; after MK-4 intervention, compared with the CCl4 group, the MK-4+CCl4 group had a significant reduction in iron accumulation. As for the measurement of iron metabolism genes in mouse liver, compared with the Control group, the CCl4 group had a significant increase in iron content, significant reductions in the mRNA expression levels of FPN and HJV, and a significant increase in the mRNA expression level of TFR1 (all P<0.05); after protection with MK-4, there was a significant reduction in iron content, significant increases in the mRNA expression levels of FPN and HJV, and a significant reduction in the mRNA expression level of TFR1 (all P<0.05). ConclusionMK-4 intervention in advance can alleviate CCl4-induced ALI in mice, possibly by inhibiting ferroptosis and improving the expression of iron metabolism-related genes in mouse liver.

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