Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

ObjectiveTo investigate the effect of laminin subunit α3 （LAMA3） on the epithelial-mesenchymal transition （EMT）， invasion， and metastasis abilities of pancreatic cancer （PC）. MethodsA comprehensive analysis was performed for tumor- and EMT-related databases to identify the EMT genes associated with PC， especially LAMA3. The methods of qRT-PCR and Western blot were used to measure the expression level of LAMA3 in PC tissue and cell lines； immunofluorescence assay was used to determine the localization of LAMA3 in PANC-1 cells； Transwell assay was used to investigate the effect of LAMA3 on the invasion and migration abilities of PC cells. The t-test was used for comparison of continuous data between groups. ResultsThe analysis of the TCGA database identified 3 EMT-related oncogenes for PC， i.e.， LAMA3， AREG， and SDC1. The LASSO-Cox regression model showed that LAMA3 had the most significant impact on the prognosis of PC （risk score=0.256 1×LAMA3+0.043 1×SDC1+0.071 4×AREG）. The Cox model and nomogram showed that the high expression of LAMA3 was an independent risk factor for the poor prognosis of PC （hazard ratio=1.32， 95% confidence interval： 1.07‍ ‍—‍ 1.62， P<0.01）. Experimental results showed that there was a significant increase in the expression of LAMA3 in pancreatic cancer tissue compared with the normal pancreatic tissue. Compared with the HPDE cell line， there were varying degrees of increase in the expression of LAMA3 in pancreatic cancer AsPC-1， BxPC-3， PANC-1， MIA PaCa-2， and SW1990 cell lines， with the highest expression level in PANC-1 cells. The enrichment analysis showed that LAMA3 was associated with the biological processes and signaling pathways such as EMT， collagen metabolism， extracellular matrix degradation， the TGF-β pathway， and the PI3K pathway. After the knockdown of LAMA3， there were significant reductions in the expression levels of N-Cadherin， Vimentin， and Snail， while there was a significant increase in the expression level of E-Cadherin. Transwell assay showed that there were significant reductions in the invasion and migration abilities of PANC-1 cells after the knockdown of LAMA3. ConclusionLAMA3 is highly expressed in PC and can promote the EMT， invasion， and migration of PC cells， and therefore， LAMA3 may be used as a novel diagnostic marker and a new therapeutic target for PC.

Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

Meningeal metastasis is a kind of serious complications of malignant tumor cells invading the leptomeninges and spreading along the cerebrospinal fluid, whose detection rate is gradually increased with advanced imaging diagnostic techniques. Although traditional intrathecal chemotherapy can prolong the survival of patients with meningeal metastasis, its efficacy is still unsatisfactory. In recent years, new intrathecal injection regimens, targeted drug therapy, and proton radiotherapy have significantly improved the prognosis of some patients with meningeal metastasis; however, these treatment regimens still require high-quality evidence-based medical evidences. This article reviews the recent advance in intrathecal injection, systemic treatments (systemic chemotherapy, molecular targeted therapy and immunotherapy), and radiotherapy of meningeal metastasis in recent years, so as to help selecting treatment regimens for meningeal metastasis in clinical practice.

Meningeal metastasis is a kind of serious complications of malignant tumor cells invading the leptomeninges and spreading along the cerebrospinal fluid, whose detection rate is gradually increased with advanced imaging diagnostic techniques. Although traditional intrathecal chemotherapy can prolong the survival of patients with meningeal metastasis, its efficacy is still unsatisfactory. In recent years, new intrathecal injection regimens, targeted drug therapy, and proton radiotherapy have significantly improved the prognosis of some patients with meningeal metastasis; however, these treatment regimens still require high-quality evidence-based medical evidences. This article reviews the recent advance in intrathecal injection, systemic treatments (systemic chemotherapy, molecular targeted therapy and immunotherapy), and radiotherapy of meningeal metastasis in recent years, so as to help selecting treatment regimens for meningeal metastasis in clinical practice.

Sucrose synthase plays a crucial role in the plant sugar metabolism pathway by catalyzing the production of uridine diphosphate (UDP)-glucose, which serves as a bioactive glycosyl donor for various metabolic processes. In this study, a sucrose synthase gene named CtSus was cloned from Cistanche tubulosa, a traditional Chinese medicine known for its rich content of diverse glycosides, based on transcriptome analysis results. The open reading frame of CtSus was 2 418 bp long encoding 805 amino acids. Sequence analysis revealed conserved domains associated with the plant sucrose synthase family at both the N-terminal and C-terminal regions of CtSus protein. Phylogenetic analysis demonstrated that CtSus shares a close evolutionary relationship with sucrose synthases from other plants within the same order. Functional identification of CtSus was performed through whole-cell catalysis coupling with UGT71BD1, a characterized glycosyltransferase enzyme. The results showed that the introduction of CtSus significantly enhanced the conversion rate of glycosylation catalyzed by UGT71BD1. The soluble expression of CtSus in Escherichia coli was further achieved using the pCold^TM TF expression vector. In vitro enzymatic assay indicated the activity of CtSus to catalyze the formation of UDP-glucose in the presence of sucrose and UDP. Real-time quantitative-polymerase chain reaction results showed that the expression patterns of CtSus gene in different parts of C. tubulosa and the suspension cell cultures of C. tubulosa under drought stress correlated with the accumulation patterns observed for phenylethanol glycosides, respectively. Furthermore, key amino acids and their interactions between enzyme and substrate were explored based on protein structure prediction and molecular docking results. Overall, our findings identify a sucrose synthase CtSus responsible for the supply of the active glycosyl donor UDP-glucose during the biosynthesis of glycoside products in C. tubulosa and have also provided a gene element that can be utilized in engineering strain construction for glycoside products production.

Objective:To analyze the normal expression levels of different lung cancer tumor markers(TM)in the cerebrospinal fluid and to explore the influence of serum TM levels and brain parenchymal metastasis,to more accurately determine whether the cerebrospinal fluid TM levels of patients with suspected meningeal metastasis is elevated.Methods:The clinical data of 80 patients diagnosed with non-lepto-meningeal metastasis at Tianjin Medical University Cancer Hospital between January 2015 and February 2024 were collected,including 16 patients without lung cancer and 64 patients with lung cancer.Normal TM levels in the cerebrospinal fluid of patients without lung cancer and the difference in TM levels between the cerebrospinal fluid and serum samples were analyzed.The correlation between serum and cerebrospinal fluid TM levels was also analyzed.We then compared the differences in TM levels in the cerebrospinal fluid between groups with brain parenchymal metastasis and without brain parenchymal metastasis.Results:Normal levels of TPSA,CA19-9,CEA,Cyfra21-1,and SCC in the cerebrospinal fluid were lower than those in the serum(P<0.05);however,the levels of ProGRP and NSE in the cerebrospinal fluid were higher than those in the serum(P<0.05).The levels of TPSA,SCC,ProGRP,NSE,CEA,CA19-9,and Cyfra21-1 in the cerebrospinal fluid did not correlate with those in the serum(all P>0.05).The cerebrospinal fluid levels of TPSA,SCC,ProGRP,and CA19-9 were not significantly increased in patients with brain parenchymal metastasis compared to those in patients without brain parenchymal metastasis(P>0.05).Al-though CEA and Cyfra21-1 levels increased(P<0.05),their median values increased by less than 2 times and were all within the reference range;whereas,the level of NSE in the group with brain parenchymal metastasis was lower than that in the control group.Conclusions:The basal levels of ProGRP and NSE in normal cerebrospinal fluid were significantly higher than those in the serum;whereas,the expression levels of other TM in the cerebrospinal fluid were significantly lower than those in the serum.Whether the levels of TM in the serum were elevated and whether brain parenchymal metastasis was present,did not have a clinically significant impact on the TM levels in the cerebrospinal fluid.

Objective The objective of this study was to analyze the long-term change trend of gastric cancer incidence in China from 1990 to 2021,and to provide scientific suggestion for the prevention and control of gastric cancer in China.Methods The incidence burden data of gastric cancer from 1990 to 2021 were obtained from the Global Burden of Disease(GBD)2021 database.The age-period-cohort model was to evaluate its independent effects,and the Nordpred model was used to predict its incidence trend from 2022 to 2031.Results The number and incidence of gastric cancer from 1990 to 2021 in China showed an upward trend,while the age-standardized incidence showed a decreasing trend.The results of age-period-cohort model showed that in the same birth cohort,the incidence of gastric cancer in China increased with age,the incidence of the total population increases from 0.91/100,000 to 233.37/100,000 in total population,the incidence of men increases from 0.91/100,000 to 508.90/100,000,and the incidence of women increases from 1.04/100,000 to 115.86/100,000.During the period from 1992-1996 to 2017-2021,the relative risk(RR)of gastric cancer incidence showed a decreasing trend with the passage of time.The RR of the total population period decreased from 1.10 to 0.72,with males decreasing from 1.06 to 0.75 and females decreasing from 1.19 to 0.65.The later the birth cohort through-out the birth cohort years,the lower the risk of onset;The RR of gastric cancer incidence in the total population decreased from 1.60 to 0.34,from 1.36 to 0.40 for males and from 2.23 to 0.22 for females.The incidence prediction results showed that by 2031,the in-cidence of gastric cancer would increase to 470,429 cases,including 323,399 cases for men and 147,029 cases for women.The stand-ardized incidence of the total population would decrease to 23.80/100,000 cases,35.13/100,000 cases for men and 24.17/100,000 cases for women.Conclusion The prevention and treatment measures of gastric cancer from 1990 to 2021 have achieved certain re-sults in reducing the risk of gastric cancer,but the incidence of gastric cancer is still serious,and it is necessary to focus on strengthe-ning the intervention for men over 50 years old.

Objective To explore the antimicrobial activity of antiparasitic drug dichlorophenol(DIC)against Staphylococcus aureus(S.aureus).Methods Antimicrobial susceptibility and resistance inducing ability of S.au-reus against DIC was detected by micro-broth dilution assay and disc diffusion test;bactericidal efficacy of DIC was assessed by time-killing curve;inhibitory effect of DIC on the formation of biofilm and eradicating of formed biofilm of S.aureus was detected by crystal violet and XTT staining;cytotoxicity of DIC was detected by cell counting kit-8(CCK-8),and a mouse model of skin abscess infection was constructed to detect the in vivo antimicrobial activity and toxicity of DIC.Results The minimal inhibitory(MIC)and minimal bactericidal concentration(MBC)of DIC against S.aureus standard strains were 2-4 μg/mL and 2-8 μg/mL,respectively.The MIC and MBC of DIC against S.aureus clinical strains were 2-8 μg/mL and 8-32 μg/mL,respectively.Disc diffusion test indicated the obvious concentration-dependent bacterial growth inhibitory effects of DIC on S.aureus standard strains.Time-killing assay revealed that DIC concentration of 4 × MIC was found to reduce the viable bacterial cells of S.aureus standard strains ATCC 29213 and ATCC 43300 from(5.51±0.27)Log10 CFU/mL and(5.44±0.08)Log10 CFU/mL to the limit of detection after 2 hours and 4 hours treatment,respectively.No drug-resistant mutant strains of S.aureus were found after 15 consecutive passage of DIC with subinhibitory concentration on bacteria.2 μg/mL DIC could significantly inhibit the formation of S.aureus biofilm and reduce the total amount of biofilm from(100±7.49)％to(11.12±2.86)％(P＜0.001).2 μg/mL DIC significantly eradicated the formed S.aureus biofilm and reduced the total biofilm from(100±10.34)％to(42.53±16.87)％(P＜0.001).DIC could significantly reduce the viable bac-terial load of S.aureus in abscess in mice,and reduce the number of viable bacteria from(9.54±0.46)Log10 CFU/abscess to(7.78±0.62)Log10 CFU/abscess(P＜0.05).Hematoxylin-eosin staining result showed that DIC could significantly reduce the abscess area and inflammatory cell infiltration in mice tissue,and was well tolerated in vivo.Conclusion DIC has low cytotoxicity and obvious in vitro and in vivo antimicrobial activity,which is expected to be an alternative treatment for drug-resistant S.aureus infection.