1.Association of liver fibrosis markers and inflammation markers with the risk of gallstones in patients with metabolic dysfunction-associated fatty liver disease
Shuai ZHANG ; Shoulu JIN ; Wanqing LI ; Xijing SHI ; Hao LIANG ; Hao DONG ; Dailong LU ; Ying ZHU ; Xiaoxing XIANG ; Jun LIU
Journal of Clinical Hepatology 2026;42(3):579-585
ObjectiveTo investigate the association of liver fibrosis scores and inflammation markers with gallstones in patients with metabolic dysfunction-associated fatty liver disease (MAFLD), as well as the mediating role of liver fibrosis scores in the relationship between inflammation markers and gallstones. MethodsA total of 14 567 patients who received physical examination and were diagnosed with MAFLD in Subei People’s Hospital from January 2014 to June 2023 were enrolled in this study, and according to the results of abdominal color Doppler ultrasound, they were divided into gallstone group with 1 724 patients and non-gallstone group with 12 843 patients. Related clinical data were collected from all patients, including demographic data, medical history, family history, physical examination, Color Doppler ultrasound, and biochemical parameters. The biomarkers associated with metabolic disorders and insulin resistance included triglyceride-glucose index (TyG), TyG-body mass index (BMI) index, atherogenic index of plasma (AIP), and non-high-density lipoprotein cholesterol-to-high-density lipoprotein cholesterol ratio (NHHR); the biomarkers associated with inflammation and nutritional status included neutrophil-to-lymphocyte ratio (NLR), neutrophil percentage-to-albumin ratio (NPAR), and monocyte-to-lymphocyte ratio (MLR); the biomarkers for assessing liver fibrosis degree and liver function included albumin-bilirubin (ALBI) score, NAFLD fibrosis score (NFS), fibrosis-4 (FIB-4) index, and aspartate aminotransferase-to-platelet ratio index (APRI). The independent-samples t test was used for comparison of normally distributed continuous data between two groups, while the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. Multivariate Logistic regression analysis, restricted cubic spline analysis, and mediating effect analysis were used to assess the association of liver fibrosis markers and inflammation markers with the risk of gallstones. ResultsThe prevalence rate of gallstones was 11.8% among the MAFLD patients. There were significant differences between the gallstone group and the non-gallstone group in sex, age, smoking history, diabetes, hypertension, lymphocytes, platelets, glucose, albumin, serum uric acid, alanine aminotransferase, aspartate aminotransferase, red blood cell, NLR, NPAR, MLR, NFS, FIB-4 index, and ALBI score (all P<0.05). The multivariate Logistic regression analysis showed that NLR (odds ratio [OR]=1.091, 95% confidence interval [CI]: 1.028 — 1.160, P<0.05), NPAR (OR=1.073, 95%CI: 1.042 — 1.105, P<0.05), MLR (OR=1.142, 95%CI: 1.057 — 1.232, P<0.05), NFS (OR=1.239, 95%CI: 1.190 — 1.291, P<0.05), and FIB-4 index (OR=1.326, 95%CI: 1.241 — 1.417, P<0.05) were influencing factors for the prevalence rate of gallstones. The restricted cubic spline analysis showed a significant non-linear association between NFS/FIB-4 index and the risk of gallstone (non-linear P<0.05). The mediating effect analysis further showed that the association of NLR, MLR, and NPAR with gallstones was partially mediated by NFS or FIB-4 index, with a mediating effect accounting for 36.79%、28.09%、29.67% and 18.31%、17.70、11.57%, respectively. ConclusionNFS and FIB-4 index have a non-linear association with the prevalence rate of gallstones in MAFLD patients, and they also mediate the association of NLR, NPAR, and MLR with the risk of gallstone.
2.Effect of thymosin β4 on a mouse model of carbon tetrachloride-induced hepatic fibrosis and its mechanism
Yunhan ZHU ; Siqi WANG ; Dengya JING ; Qinying FENG
Journal of Clinical Hepatology 2026;42(3):593-599
ObjectiveTo investigate the therapeutic effect and potential mechanism of thymosin β4 (Tβ4) on carbon tetrachloride (CCl4)-induced hepatic fibrosis by regulating the expression of platelet-derived growth factor (PDGF) and inducing the apoptosis of hepatic stellate cell (HSC), and to provide new experimental evidence for anti-hepatic fibrosis treatment in clinical practice. MethodsA total of 30 male C57 mice were randomly divided into normal control group, model group, low-dose Tβ4 treatment group (3 mg/kg), middle-dose Tβ4 treatment group (6 mg/kg), and high-dose Tβ4 treatment group (12 mg/kg), with 6 mice in each group. The mice in the normal control group were fed with a normal diet ad libitum, and those in the other groups were given intraperitoneal injection of 50% CCl4 mixed with olive oil to establish a model of hepatic fibrosis. After successful modeling confirmed by ultrasound and histopathology, the mice in each treatment group were given subcutaneous injection of Tβ4 for 4 consecutive weeks. Liver tissue was collected at the end of the experiment, and HE staining and Masson staining were used to observe histopathological changes; quantitative real-time PCR was used to measure the mRNA expression level of PDGF; TUNEL assay was used to assess the apoptosis of HSC. 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 model group, the middle- and high-dose Tβ4 treatment groups had varying degrees of alleviation of hepatic fibrosis. Quantitative real-time PCR showed that Tβ4 could significantly downregulate the mRNA expression level of PDGF in liver tissue, with a significant difference between the treatment groups (P>0.05), and there was no significant difference in the mRNA expression level of PDGF between the high-dose Tβ4 treatment group and the normal control group (P>0.05). TUNEL assay showed that the middle- and high-dose Tβ4 treatment groups had a significantly higher number of apoptotic HSCs than the model group. ConclusionTβ4 may improve CCl4-induced hepatic fibrosis in mice by downregulating the expression of PDGF and promoting the apoptosis of HSC, suggesting that it has a potential application value in the treatment of hepatic fibrosis.
3.Effect of astragaloside Ⅳ on a mouse model of carbon tetrachloride-induced liver fibrosis and its mechanism
Wanchun ZHU ; Jiahao QIU ; Yu CUI ; Yijing ZHANG ; Zhi SHANG ; Yueqiu GAO ; Lingying HUANG
Journal of Clinical Hepatology 2026;42(3):608-617
ObjectiveTo investigate the liver-protecting and anti-liver fibrosis effects of astragaloside Ⅳ (AS-Ⅳ) in vitro and in vivo, as well as its mechanism of action in intervention against liver fibrosis. MethodsIn the animal experiment, C57BL/6J mice were divided into control group, model group, low-dose AS-Ⅳ (20 mg/kg) group, and high-dose AS-Ⅳ (80 mg/kg) group. The mice were given intraperitoneal injection of carbon tetrachloride for 6 weeks to induce liver fibrosis, and since week 3 of injection, the mice in the low-dose AS-Ⅳ group and the high-dose AS-Ⅳ group were given AS-Ⅳ by gavage at a dose of 20 mg/kg and 80 mg/kg, respectively. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured after 4 weeks of administration, as well as the serum levels of hyaluronic acid (HA), laminin (LN), procollagen Ⅲ N-terminal peptide (PⅢNP), and collagen type Ⅳ (Col-Ⅳ). HE staining, picrosirius red staining, and Masson staining were used to observe liver histopathology and collagen deposition; RT-qPCR was used to measure the mRNA expression levels of Acta2, Col1a1, and Col3a1 in liver tissue, and Western blot was used to measure the protein expression levels of α-smooth muscle actin (α-SMA), collagen type Ⅲ (Col-Ⅲ), phosphatidylinositol 3-kinase (PI3K), phosphorylated PI3K (pPI3K), protein kinase B (Akt), and phosphorylated AKT (p-Akt) in liver tissue; transcriptome sequencing was performed for liver tissue to identify differentially expressed genes and perform a bioinformatics analysis. In the cell experiment, transforming growth factor-β (TGF-β) was used to induce the activation of LX-2 cells, and the PI3K inhibitor LY294002 and the PI3K activator 740 Y-P were used for intervention. The cells were divided into control group, model group, AS-Ⅳ group, LY294002 group, and AS-Ⅳ+740 Y-P group, and the cells were harvested after 36 hours of intervention. Changes in the protein expression levels of α-SMA, Col-Ⅲ, pPI3K/PI3K, and pAkt/Akt in LX-2 cells were measured, as well as changes in the relative mRNA expression levels of Acta2, Col1a1, and Col3a1. 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 animal experiment, compared with the model group, the AS-Ⅳ treatment group had significant reductions in the serum levels of ALT, AST, HA, LN, PⅢNP, and Col-Ⅳ (all P<0.01), the mRNA expression levels of Acta2, Col1a1, and Col3a1 in liver tissue (all P<0.05), and the protein expression levels of α-SMA, Col-Ⅲ, pPI3K, and pAkt (Ser473) in liver tissue (all P<0.05). In the cell experiment, compared with the control group, the model group had significant increases in the protein expression levels of α-SMA, Col-Ⅲ, pPI3K, and pAkt (Ser473) after TGF-β induction (all P<0.05); compared with the model group, the AS-Ⅳ group had significant reductions in the protein expression levels of α-SMA, Col-Ⅲ, pPI3K, and pAkt (Ser473) (all P<0.05), and both the AS-Ⅳ group and the LY294002 group had significant reductions in the protein expression level of pPI3K and the relative mRNA expression levels of Acta2, Col1a1, and Col3a1 (all P<0.05). Compared with the AS-Ⅳ group, there were significant increases in the protein expression level of pPI3K and the relative mRNA expression levels of Acta2, col1a1, and Col3a1 after 740 Y-P intervention (all P<0.05). ConclusionAS-Ⅳ can inhibit hepatic stellate cell activation and improve liver fibrosis, possibly by inhibiting the PI3K/Akt signaling pathway.
4.Risk factors for lower extremity deep vein thrombosis in patients with acute necrotizing pancreatitis and effectiveness of risk assessment models
Liu YANG ; Gang ZHU ; Longfei ZENG ; Benjian GAO ; Bo LI
Journal of Clinical Hepatology 2026;42(3):647-654
ObjectiveTo investigate the influencing factors and independent risk factors for lower extremity deep vein thrombosis (DVT) in patients with acute necrotizing pancreatitis (ANP), to analyze the effectiveness of three commonly used risk assessment models for thrombosis (Caprini score, Padua score, and Wells score), and to provide a reference for clinical identification of high-risk individuals and optimization of prevention and treatment strategies. MethodsA retrospective analysis was performed for the clinical data of 320 patients with ANP who were admitted to Luzhou People’s Hospital and The Affiliated Hospital of Southwest Medical University from April 2013 to April 2024, and according to the presence or absence of DVT during hospitalization, the patients were divided into thrombosis group with 25 patients and control group with 295 patients. After propensity score matching, the two groups were compared in terms of past history and various examination results during hospitalization. The risk factors for lower extremity DVT in ANP patients during hospitalization were analyzed through univariate and multivariate Logistic regression, and a DVT risk prediction model was established based on independent influencing factors. The receiver operating characteristic (ROC) curve was used to assess the performance of models, and the DeLong test was used for comparison of the area under the ROC curve (AUC), sensitivity, and specificity. The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups; the chi-square test was used for comparison of categorical data between groups. ResultsAfter matching, the patients were divided into thrombosis group with 24 patients and control group with 112 patients. The clinical characteristics analysis showed that compared with the control group, the thrombosis group had significantly higher degree of pancreatic necrosis, D-dimer level, Bedside Index for Severity in Acute Pancreatitis (BISAP) score, and proportion of patients undergoing dialysis (all P<0.05). The multivariable Logistic regression analysis showed that BISAP score, degree of pancreatic necrosis, and D-dimer level were independent risk factors for lower extremity DVT in ANP patients during hospitalization (all P<0.05). The BISAP-Caprini score model had an AUC of 0.832 (95% confidence interval: 0.722 — 0.942, P<0.001) in predicting the risk of lower extremity DVT, with a Youden index of 1.661, an optimal cut-off value of 0.26, a sensitivity of 75.0%, and a specificity of 91.1%. ConclusionD-dimer, BISAP score, and the degree of pancreatic necrosis are independent risk factors for lower extremity DVT in patients with ANP during hospitalization, and the BISAP-Caprini score model can effectively predict the risk of DVT in ANP patients.
5.Regulatory role and mechanism of intrahepatic macrophages in metabolic dysfunction-associated fatty liver disease
Journal of Clinical Hepatology 2026;42(3):668-675
With the improvement of living standards, the incidence rate of metabolic dysfunction-associated fatty liver disease (MAFLD) is gradually increasing with a younger age of onset, and MAFLD has become a global health problem, while its specific pathogenesis remains unclear. Macrophages, as one of the important cells involved in the pathogenesis of MAFLD, have the ability to present antigens, eliminate pathogenic microorganisms, and promote liver inflammatory responses, thereby attracting wide attention for a long time. The latest studies have shown that macrophages may become a new therapeutic target for MAFLD. This article systematically reviews the role of intrahepatic macrophages in liver inflammation caused by MAFLD, including their activation, polarization, recruitment mechanisms, and interactions with other cells, in order to provide new ideas and perspectives for the clinical prevention and treatment of MAFLD.
6.LI Guolie's Experience in Treating Orthostatic Hypotension from the Perspective of "Raising the Clear and Directing the Turbid Downward"
Zhihui JIN ; Yanjie JIANG ; Wenshan LI ; Xiaoyu ZHU ; Yan LU ;
Journal of Traditional Chinese Medicine 2026;67(9):941-945
This paper summarizes professor LI Guolie's clinical experience in treating orthostatic hypotension (OH) based on the theory of "raising the clear and directing the turbid downward". It is considered that the core pathogenesis of OH lies in the body's transition from a supine to an upright position, during which dysfunction of the middle jiao (焦) transformation and transportation, along with impaired pivot function, hinders the ascending of clear yang and the descending of turbid yin. Treatment should follow the general principle of "ascending the clear and directing the turbid downward", placing emphasis on distinguishing the primary and secondary aspects. For cases where the clear yang fails to ascend, the self-formulated Li's Shengqing Jiangzhuo Decoction (李氏升清降浊汤)is used to supplement qi, raise the clear, and strengthen the middle jiao. For cases where the turbid yin fails to descend, the self-formulated Wuxiang Qingzhuo Beverage(五香清浊饮)with modifications is applied to resolve phlegm, eliminate stasis, harmonize the middle, and descend the turbid.
7.Similarities and Differences between Myofascial Trigger Points and Sinew Knot:from the Perspective of the Channel Sinew Theory
Zijie CHEN ; Haohan ZHU ; Xingke SONG ; Xue PU ; Junying WANG
Journal of Traditional Chinese Medicine 2026;67(9):1023-1026
Myofascial trigger points, as hyperirritable spots within taut bands of skeletal muscle, can induce local or referred pain, and show a high degree of similarity to acupoints in traditional Chinese medicine (TCM), particularly the so-called sinew knot lesion point. From the perspective of channel sinew theory, and by examining the correlations of myofascial trigger points with acupoints and channel sinew disorders, this study aims to compare the similarities and differences between MTrPs and sinew knot lesion points in terms of pathological mechanisms, needling analgesic mechanisms, and therapeutic approaches. The goal is to deepen the understanding of MTrPs and dry needling, and provide a modern scientific perspective on channel sinew theory and the sinew knot lesion point.
8.Study on the safety and efficacy of micro-perfusion device for preserving isolated porcine limbs
Pengkai LI ; Zhaodi MI ; Shen LI ; Man YUAN ; Xiwei PENG ; Jia LÜ ; Sice WANG ; Zhibo JIA ; Xiangyu SONG ; Yixuan ZHU ; Chonghui LI ; Moling XIAO ; Wenjing XU ; Jiang PENG
Organ Transplantation 2026;17(3):422-431
Objective To evaluate the safety and efficacy of a self-developed micro-normothermic machine perfusion (NMP) system (micro-perfusion device) for preserving isolated porcine limbs. Methods Five healthy Landrace pigs were selected, and their left and right forelimbs were randomly divided into the NMP group and static cold storage (SCS) group. The NMP group was perfused with the self-developed micro-perfusion device and polymerized hemoglobin perfusate for 32 hours at normothermia, while the SCS group was preserved at 4 ℃. Hemodynamic parameters such as perfusion pressure and flow were monitored. The pH value, partial pressure of oxygen (PO2), lactic acid (Lac), creatine kinase (CK) and lactate dehydrogenase (LDH) in the perfusate were measured. Hematoxylin-eosin staining was used to assess the muscle tissue structure, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was employed to evaluate muscle cell apoptosis, and immunohistochemistry staining was applied to detect the expressions of tumor necrosis factor (TNF)-α and interleukin (IL)-6. A mixed-effects model was used to analyze the effects of time and treatment methods on tissue structure, cell apoptosis and inflammatory factors. Results The device could stably maintain a perfusion pressure of (69±15) mmHg and a flow rate of (117±42) mL/min. The pH value and electrolytes of the perfusate were generally stable, with PO2 maintained at a high level. Lac was maintained at 5.38(3.81, 6.45) mmol/L, while CK and LDH increased over time. After 32 hours of perfusion in the NMP group, both the myocyte spacing and apoptosis rate were better than those in the SCS group. Mixed-effects model analysis showed that there were statistically significant differences in the effects of NMP treatment and SCS treatment on myocyte spacing and apoptosis rate per unit time (both P < 0.05). There were no statistically significant differences in TNF-α and IL-6 between the two groups, and mixed-effects model analysis showed no statistically significant differences in the effects of NMP treatment and SCS treatment on TNF-α and IL-6 per unit time (both P > 0.05). Conclusions The micro-perfusion device used in this study may achieve 32-hour normothermic preservation in a porcine limb amputation model, maintain basic metabolism and ionic homeostasis, reduce muscle structural damage and cell apoptosis without inducing additional inflammatory responses. This technology is expected to significantly extend the time window for replantation of amputated limbs in disaster rescue and long-distance transportation, providing an important technical basis for clinical translation and subsequent replantation research.
9.Risk prediction of post-reperfusion syndrome during adult living donor liver transplantation based on machine learning
Yuanwei WANG ; Yulong ZHU ; Ling XU
Organ Transplantation 2026;17(3):460-468
Objective To compare multiple models and analyze SHAP feature contributions to construct a machine learning risk prediction model for post-reperfusion syndrome (PRS) during adult living donor liver transplantation (LDLT). Methods Clinical data of 390 patients who underwent LDLT due to end-stage liver disease at the First Affiliated Hospital of Anhui Medical University from May 2023 to April 2025 were collected. Five machine learning models, including random forest, logistic regression, XGBoost, decision tree, and AdaBoost, were compared. Feature selection was performed using Lasso regression, and the training set was divided into five folds for stratified cross-validation. The generalization ability of the model was evaluated on the independent test set. The models were comprehensively compared based on key evaluation indicators such as recall rate, accuracy rate, precision rate, area under the curve (AUC) and F1 value, to determine the optimal model. Results Eight potential factors for predicting PRS were selected. The random forest model demonstrated the best prediction performance in both the training set and the test set. Its accuracy rate was as high as 84.2% (AUC = 0.894, 95% confidence interval 0.808-0.964) in the test set. The importance ranking of PRS predictors was determined through SHAP value analysis. Cold ischemia time, K+ during the anhepatic period and the model for end-stage liver disease (MELD) score were all identified as the most significant predictors. Conclusions Based on eight key indicators including cold ischemia time, portal vein occlusion time, pre-reperfusion body temperature, alkaline reserve during the anhepatic period, K+ during the anhepatic period, MELD score, left ventricular end-diastolic diameter and graft volume to standard liver volume ratio, the PRS prediction model constructed using the random forest algorithm demonstrates the best prediction performance in the test set, providing certain assistance for subsequent clinical predictions.
10.Research on the dual-pathway construction of organ donation incentive mechanism: delayed rewards and timely compensation
Chang YAO ; Hanle WANG ; Zhelin ZHU
Organ Transplantation 2026;17(3):476-485
Exploring the institutional innovation paths for the incentive mechanism of organ donation in our country, with "delayed rewards" and "timely compensation" as the core logic, starting from the dilemma of public goods supply and the theory of incentive compatibility, a reward system that balances ethical bottom lines and policy effectiveness is constructed. Firstly, reviewing international experiences reveals that Israel's point-based priority system, Singapore's "presumed consent + financial neutrality" model, the indirect incentive mechanism in the United States, and the family reciprocal incentive model in Japan all demonstrate a balanced institutional equilibrium between legal legitimacy and cultural compatibility. Secondly, analyzing the threefold predicaments of current incentives in our country, namely, the empty circulation of incentives, the absence of compensation and the lack of credibility. To solve the policy predicament of "establishing but difficult to implement" in organ transplantation work, this paper proposes a "institutional investment" analytical framework, emphasizing the design of dual paths of delayed reward type (future preferential treatment, point-based system) and timely compensation type (economic assistance, fund feedback) to form a positive cycle of incentives and institutional trust. Finally, this paper suggests establishing an organ donation incentive system supported by a unified point-based system, a national compensation fund and a third-party supervision system, to achieve the policy vision of "paying tribute to kindness in incentives and extending life through the system".

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