ObjectiveThis paper aims to investigate the effects and mechanism of Mimenghua prescription in modulating the mitogen-activated protein kinase kinase （MEK）/rat sarcoma viral oncogene homolog （Ras）/rapidly accelerated fibrosarcoma kinase （Raf）/extracellular signal-regulated kinase （ERK） signaling pathway to inhibit inflammatory responses in a dry eye animal model. MethodsA total of 60 C57BL/6J mice （eight weeks old， half male and half female） were used in the experiment. Ten mice were randomly selected as the blank control group， while the remaining 50 were exposed to a controlled dry system and received instillation of 0.2% benzalkonium chloride （BAC） into the eyes for four weeks to establish a dry eye mouse model. After successful modeling， the mice were randomly divided into five groups： Model group， sodium hyaluronate group， and Mimenghua prescription groups with low dose （4.83 g·kg^-1）， medium dose （9.67 g·kg^-1）， and high dose （19.34 g·kg^-1）. The mice in the model group received an equal volume of normal saline via gavage for four weeks. The mice in the sodium hyaluronate group received instillation of sodium hyaluronate eye drops twice daily for 14 consecutive days. The tear secretion volume， tear film break-up time （TBUT）， and corneal fluorescein staining were evaluated once every two weeks. After four weeks of administration， mice were euthanized， and their lacrimal gland tissues and corneas were harvested. Hematoxylin-eosin （HE） staining was used to assess histopathological morphology. Western blot was performed to detect the protein expression levels of MEK， Ras， Raf， and ERK. Enzyme-linked immunosorbent assay （ELISA） was used to measure the contents and expressions of MEK， Ras， Raf， ERK， and interleukin （IL）-1β in lacrimal gland and corneal tissues of the mice in each group. Quantitative real-time polymerase chain reaction （Real-time PCR） was employed to determine mRNA expression levels of MEK， Ras， Raf， and ERK. ResultsThe Mimenghua prescription groups and the sodium hyaluronate group exhibited significantly increased tear secretion volume （P<0.05） and prolonged TBUT （P<0.05） after treatment. Ocular surface damage of mice was visibly recovered. Western blot results indicated that protein expression levels of MEK， Ras， Raf， and ERK in the lacrimal gland and corneal tissues were significantly downregulated in the sodium hyaluronate group and Mimenghua prescription group with high dose （P<0.05）. ELISA results showed that IL-1β levels were highest in the model group but significantly reduced in the sodium hyaluronate group and Mimenghua prescription groups （P<0.05）. Both ELISA and Real-time PCR results demonstrated that the expression levels of MEK， Ras， Raf， and ERK in the lacrimal glands and corneal tissues were significantly elevated in the model group （P<0.05）， but markedly downregulated in the sodium hyaluronate group and Mimenghua prescription groups （P<0.05）， suggesting that Mimenghua prescription can decrease the expressions of MEK， Ras， Raf， and ERK in the lacrimal glands and corneal tissues. ConclusionMimenghua prescription can reduce inflammatory responses， increase tear secretion， prolong TBUT， and promote corneal recovery by inhibiting the MEK， Ras， Raf， and ERK signaling pathways in lacrimal gland and corneal tissues.

ObjectiveTo investigate the association between noninvasive liver fibrosis markers and carotid plaque （CP） in patients with lean metabolic dysfunction-associated fatty liver disease （MAFLD）， and to provide a basis for screening high-risk populations. MethodsA total of 957 patients with lean MAFLD who underwent physical examination in Subei People’s Hospital from January 2021 to June 2023 was enrolled as the observation cohort， with the presence or absence of CP as the outcome， and fibrosis-4 （FIB-4） index and nonalcoholic fatty liver disease fibrosis score （NFS） were used to assess liver fibrosis degree. The independent-samples t test was used for comparison of normally distributed continuous data between two groups， and 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. The multivariate logistic regression analysis， the restricted cubic spline analysis， the receiver operating characteristic curve， and the mediation effect analysis were used to investigate the association between liver fibrosis degree and CP. ResultsThe prevalence rate of CP was 36.6% in the lean MAFLD population. Compared with the non-CP group（n=607）， the CP group （n=350） had a significantly higher proportion of male patients， a significantly higher proportion of patients with smoking/diabetes/hypertension， and significantly higher levels of age， creatinine， blood urea nitrogen， triglycerides， fasting blood glucose， aspartate aminotransferase， aspartate aminotransferase/alanine aminotransferase ratio， NFS， and FIB-4 index， as well as significantly lower levels of platelet count and albumin （all P<0.05）. The multivariate logistic regression analysis showed that after adjustment for confounding factors， FIB-4 index （odds ratio［OR］=2.979， 95% confidence interval［CI］：2.141 — 4.219， P<0.001） and NFS （OR=1.747， 95%CI： 1.499 — 2.046， P<0.001） were positively correlated with CP. Both FIB-4 index and NFS had a good value in predicting CP. Hypertension had a significant indirect effect on the prevalence rate of CP through its impact on liver fibrosis markers， and its mediating effect accounted for 39.5% — 40.8% of the total effect （P<0.001）. ConclusionIn patients with lean MAFLD， NFS and FIB-4 index are significantly positively correlated with the prevalence rate of CP， and they can be used as potential epidemiological predictive indicators. Liver fibrosis markers may play a mediating role in the association between hypertension and CP. Interventions targeting hypertension and liver fibrosis markers may help to prevent and delay the progression of CP.

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.

Background/Aims: Liver transplantation (LT) following total hepatectomy is a life-saving treatment for hepatocellular carcinoma (HCC). The HCC recurrence after LT hinders the effectiveness of the procedure. The objective of this study is to develop a pre-operative risk stratification model based on a liquid biopsy. Methods: We conducted a comprehensive multi-omics study of 260 HCC patients from three centers, including clinical data, low-coverage whole-genome sequencing of cell-free DNA (cfDNA) from plasma, as well as whole-exome, single-nucleus RNA, and spatial transcriptomics from matched tumor and non-tumor tissues. Results: We identified cfDNA-derived copy number alteration (CNA) signatures associated with post-transplant recurrence. By integrating cfDNA-derived CNA profiles with single-cell transcriptomic data, we traced recurrence-associated cfDNA to a distinct subpopulation of malignant cells within the primary tumor. These cells were embedded in a pro-metastatic microenvironment of specialized endothelial subtypes and cancer-associated fibroblasts. Notably, most recurrence-associated lesions were detectable in cfDNA prior to liver transplantation (LT). Building on these insights, we developed the ZJU Criteria based on CNA fragments and tumor markers, a pre-LT risk prediction tool that integrates conventional clinical factors with cfDNA-derived CNA signatures, and validated it using internal and independent external cohorts. Conclusion Our findings suggest that post-transplant recurrence commonly originates from advanced subclones that emerge late during tumor evolution. The ZJU Criteria provides an accurate, non-invasive strategy that significantly improves pre-LT risk stratification and clinical decision-making for patients with HCC.

Objective To explore the clinical efficacy of biplane double-supported screw fixation(BDSF)in the treatment of femoral neck fracture(FNF)and the influencing factors of surgical failure.Methods A total of 360 patients with FNF hospitalized in our hospital from November 2022 to May 2024 were selected as the research objects.According to the random number table method,the patients were divided into the observation group[application of BDSF under the multi-disciplinary treatment(MDT)and green channel mode]and the control group[application of cannulated compression screw(CCS)fixation under the MDT and green channel mode],with 180 cases in each group;According to the failure of surgical treatment or fixation in the observation group,the patients were further divided into the surgical failure group(n=50)and effective surgery group(n=130).The risk factors were analyzed by multivariate Logistic regression.The establishment and fitting of the model were analyzed by Logistic regression.The discrimination,prediction accuracy and clinical value of the model were evaluated by receiver operating characteristic(ROC)curve,calibration curve and decision curve analysis(DCA)curve,respectively.Results Compared with the control group,the observation group had shorter operation time and hospitalization time(P<0.05),less intraoperative blood loss and fewer intraoperative fluoroscopy times(P<0.05),lower proportion of patients with avascular necrosis of femoral head and fixation failure(P<0.05),and higher Harris hip score 24 months after operation(P<0.05).Garden classification,fracture line location,reduction quality and preoperative traction were the independent influencing factors of surgical failure(P<0.05).The ROC curve analysis showed that the area under the curve(AUC)of the model to predict the risk of surgical failure was 0.867(95%CI:0.751 to 0.946).The calibration curve analysis results showed that the predicted probability of the model was approximately equal to the actual probability,with a Brier value of 0.095.DCA analysis results showed that the threshold probability of the model was 0.1 to 0.9,indicating a high net profit.Conclusion BDSF has a good effect in FNF surgery,which can shorten the operation time and hospitalization time,reduce the number of intraoperative fluoroscopy and bleeding,which is more conducive to hip joint function recovery.Garden classification,fracture line location,reduction quality,and preoperative traction are all the independent influencing factors for surgical failure.

Primary aldosteronism(PA)is the most common cause of secondary hypertension,yet its diagnosis remains under-recognized,with a high rate of missed diagnoses.This condition significantly impacts multiple systems,including the cardiovascular,renal,metabolic,and skeletal systems,resulting in notable complications.Patients with Klinefelter syndrome(KS)exhibit a markedly higher prevalence of metabolic disorders,which may further exacerbate cardiovascular and endocrine dysfunction.Here we report a patient who presented with refractory hypertension and an incidentally discovered adrenal mass,and was ultimately diagnosed with a left adrenal aldosterone-producing adenoma combined with KS.We provide a detailed description of the patient's clinical manifestations,biochemical indices,diagnostic process,and postoperative histopathological findings in order to enhance the understanding of PA and KS and their potential interconnections,which offers diagnostic insights to reduce misdiagnosis and missed diagnoses.

AIM:This study aimed to investigate the inhibitory effects and potential mechanisms of cyasterone(CYA)on inflammation and apoptosis of chondrocytes in knee osteoarthritis(KOA).METHODS:A rat model of KOA was established through anterior cruciate ligament transection(ACLT).Rats were randomly divided into 5 groups(n=6 in each group):normal control(NC),ACLT,ACLT+CYA(10 mg/kg),ACLT+CYA+SR9009(BMAL1 inhibitor),and ACLT+CYA+LY294002(PI3K inhibitor).Pathological changes in cartilage were evaluated using hematoxylin-eosin(HE)and safranin O/fast green staining,graded according to the Osteoarthritis Research Society International(OARSI)system.Immunohistochemistry was employed to measure the expression levels of BMAL1,phosphorylated phosphati-dylinositol 3-kinase(p-PI3K),phosphorylated protein kinase B(p-AKT)and phosphorylated nuclear factor-κB(p-NF-κB)in cartilage.An in vitro KOA model was created by stimulating rat chondrocytes with interleukin-1β(IL-1β).Cell vi-ability was assessed using the CCK-8 assay,while enzyme-linked immunosorbent assay(ELISA)was used to quantify pro-inflammatory cytokines(IL-6,IL-1β and TNF-α).Apoptosis was analyzed via flow cytometry,and the protein expression levels of BMAL1,PI3K,p-PI3K,AKT,p-AKT,NF-κB and p-NF-κB were evaluated using Western blot.RESULTS:In vivo,the rats in ACLT group exhibited significant chondrocyte degradation and lacunae formation compared with NC group,confirming the successful establishment of the KOA model.The rats in ACLT+CYA,ACLT+CYA+SR9009 and ACLT+CYA+LY294002 groups showed improved cartilage integrity compared with ACLT group,with reduced OARSI scores(P<0.05),increased BMAL1 expression(P<0.05),and decreased levels of p-PI3K,p-AKT and p-NF-κB(P<0.05).In vitro,the chondrocytes in IL-1β+CYA,IL-1β+CYA+SR9009 and IL-1β+CYA+LY294002 groups exhibited lower levels of IL-6,IL-1β and TNF-α(P<0.05),decreased apoptosis rates(P<0.01),increased BMAL1 protein ex-pression(P<0.05),and reduced p-PI3K/PI3K,p-AKT/AKT and p-NF-κB/NF-κB ratios(P<0.05),compared with IL-1β group.CONCLUSION:Cyasterone alleviates chondrocyte inflammation and apoptosis in KOA by activating BMAL1,which subsequently inhibits the phosphorylation of PI3K/AKT/NF-κB signaling pathway.

Uric acid is the end product of purine metabolism in the human body. In recent years, the role of uric acid in female fertility and assisted reproductive technology (ART) has gained increasing attention. Dysregulation of uric acid metabolism can lead to hyperuricemia (HUA). HUA is not only closely related to metabolic syndrome and cardiovascular diseases but may also adversely affect female fertility by influencing ovarian function and embryos development. In this review, we explored the role of uric acid in female fertility, including its association with female subfertility, infertility, adverse pregnancy outcomes and metabolic syndrome, as well as its potential impact on ART like in vitro fertilization-embryo transfer and intracytoplasmic sperm injection. Further studies are needed to clarify the threshold and clinical intervention value of uric acid levels in women of childbearing age, providing a basis for reproductive health counseling and personalized pregnancy assistance for HUA patients of childbearing age.

Objective:To explore the impact of previous cytomegalovirus (CMV), herpes simplex virus (HSV), and rubella virus (RV) infection on pregnancy outcomes in infertile women undergoing the first in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET) treatment. Methods:A retrospective cohort study was conducted to analyze the clinical data of women who underwent IVF/ICSI-ET for the first time at the Reproductive Medicine Center, the First Affiliated Hospital of Zhengzhou University from December 2017 to December 2022. The patients were divided into CMV-IgG (+) group ( n=154), RV-IgG (+) group ( n=86), HSV-IgG (+) group ( n=93) and IgG all-negative group ( n=172). The pregnancy outcomes of the patients in the virus-only infection group and the IgG all-negative group were compared. Patients who were previously co-infected with CMV and HSV were classified as the CMV+HSV-IgG (+) group ( n=344), and the pregnancy outcomes of patients with previous CMV and HSV co-infection and those with infection alone were further compared. Results:The two pronuclei (2PN) fertilization rate [63.90% (1 195/1 870)], the clinical pregnancy rate [51.30% (79/154)], and the live birth rate [45.45% (70/154)] of the CMV-IgG (+) group were significantly lower than those of the IgG completely negative group [68.68% (1 469/2 139), P=0.001; 68.60% (118/172), P=0.001; 61.05% (105/172), P=0.005]. The 2PN fertilization rate [61.62% (729/1 183)], the clinical pregnancy rate [50.54% (47/93)], and the live birth rate [43.01% (40/93)] of the HSV-IgG (+) group were significantly lower than those of the IgG completely negative group [68.68% (1 469/2 139), P=0.001; 68.60% (118/172), P=0.004; 61.05% (105/172), P=0.005]. There were no statistical differences in the 2PN fertilization rate, the clinical pregnancy rate, and the live birth rate between the RV-IgG (+) group and the IgG completely negative group (all P>0.05). Compared with the IgG completely negative group, there were no significant differences in the risk of complications such as gestational diabetes, hypertensive disorders of pregnancy and neonatal outcomes in the CMV-IgG (+) group, RV-IgG (+) group, and HSV-IgG (+) group (all P>0.05). Multivariate logistic regression analysis showed that CMV-IgG (+) ( OR=0.453, 95% CI: 0.280-0.734, P=0.001; OR=0.515, 95% CI: 0.321-0.825, P=0.006), HSV-IgG (+) ( OR=0.425, 95% CI: 0.245-0.738, P=0.002; OR=0.447, 95% CI: 0.259-0.771, P=0.004) and CMV+HSV-IgG (+) ( OR=0.491, 95% CI: 0.329-0.733, P=0.001; OR=0.528, 95% CI: 0.357-0.780, P=0.001) were all independent influencing factors of patients' clinical pregnancy and live birth. There were no statistical differences in the clinical outcomes between the previous CMV and HSV co-infection group and the single infection group ( P>0.05). Conclusion:Previous CMV or HSV infection alone reduced the fertilization rate, the clinical pregnancy rate and the live birth rate of patients undergoing IVF/ICSI-ET treatment, but had no significant impact on pregnancy complications and neonatal outcomes. Pregnancy outcomes of patients with previous CMV and HSV co-infection were similar to those with infection alone.

A mounting body of research suggests that circRNAs significantly contribute to the develop-ment of myocardial fibrosis.The microarray results of human circular RNA expression profile indicated that circHERC4_041 expression increased in the myocardium of patients with heart failure,RT-qPCR a-nalysis confirmed that the myocardial expression level of circHERC4_041 in individuals with heart failure were considerably elevated compared to that in healthy organ donors.Fluorescence in situ hybridization(FISH)confirmed that circHERC4_041 was abundant in the cytoplasm of human cardiomyocyte AC16.Overexpression of circHERC4_041 in mouse myocardial fibroblasts(mCFs)mediated by adenovirus in-hibited the expression of fibrosis-related proteins in mCFs.Experiments involving cell proliferation,wound healing,and Transwell assays demonstrated that overexpression of circHERC4_041 suppressed the growth and mobility of mCFs(P＜0.001).Sequence analysis results suggested that circHERC4_041 con-tains potential ribosome entry sequence(IRES)and open reading frame(ORF).Western blot confirmed that circHERC4_041 could translate the 516 amino acid HERC4-516aa protein,which was mainly located in the cytoplasm of the cell.Cell functional experiments confirmed that circHERC4_041 inhibited the fi-brotic phenotype of mCFs by specifically translating HERC4-516aa(P＜0.05).The specific interaction between HERC4-516aa and transglutaminase 2(TGM2)was confirmed by IP-MS screening and Co-IP i-dentification.Further results found that the degradation of TGM2 was promoted through proteasome path-way.The overexpression of TGM2 in mCFs facilitated by adenoviral vectors could counteract the suppres-sive effects of HERC4-516aa on the fibrotic phenotype of mCFs.Therefore,this study confirmed that the HERC4-516aa protein translated by circHERC4_041 can specifically bind to TGM2 to inhibit the fibrotic phenotype of myocardial fibroblasts.