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.Etiological surveillance for influenza-like illness cases in Jiangsu Province
SHI Chunlei ; DAI Qigang ; DONG Yanhui ; LIU Dongsheng ; ZHOU Shengnan
Journal of Preventive Medicine 2026;38(2):109-114
Objective:
To analyze the etiological surveillance results of influenza-like illness (ILI) cases in Jiangsu Province, and investigate the distribution characteristics of different influenza virus types, so as to provide the evidence for improving influenza prevention and control measures.
Methods:
Influenza laboratory testing data for sentinel surveillance of ILI cases in Jiangsu Province from 2019 to 2024 were collected through the China Influenza Surveillance Information System. The positive detection rate of influenza virus was calculated, and descriptive analysis was performed to characterize the distribution of different influenza virus types. Using the farthest neighbor linkage method, influenza virus positive detection rates clustering was analyzed by year and week. Clusters were defined based on inter-cluster distance, and the intensity of the positive detection rate was visualized through color gradients in the clustering heatmap.
Results:
From 2019 to 2024, a total of 183 878 ILI specimens were collected in Jiangsu Province. Among them, 20 059 specimens tested positive for influenza virus, corresponding to an overall positive detection rate of 10.91%, and an average annual positive detection rate of 10.89%. The primary circulating influenza virus types were influenza A H3N2 subtype, accounting for 40.92%, followed by influenza B Victoria linage at 34.00%, and influenza A H1N1 subtype at 24.80%. Influenza B Yamagata linage was not detected throughout the five-year period. Influenza A H3N2 subtype predominated during two distinct periods: from January to March 2019, and from June 2022 to December 2023. Influenz B Victoria linage was the dominant type from April 2019 to May 2022 and again from January to April 2024. Influenza A H1N1 subtype emerged as the primary type from May to December 2024. Year-based clustering analysis grouped the annual positive detection rates from 2019 to 2024 into three clusters. The closest cluster distance was observed between 2019 and 2024. The highest annual positive detection rate occurred in 2023. Both influenza A H3N2 and H1N1 subtype each formed a single cluster, with their peak positive detection rates also recorded in 2023. Influenza B Victoria lineage was separated into two clusters, with its highest positive detection rate occurring in 2020. Week-based clustering analysis revealed that influenza virus detection was concentrated in weeks 47 to 52 and weeks 1 to 15. More specifically, the positive detection rates for influenza A H3N2 subtype peaked during weeks 30 to 34 and weeks 42 to 52; for influenza A H1N1 subtype, during weeks 9 to 15 and weeks 51 to 52; and for influenza B Victoria lineage, during weeks 1 to 11 and weeks 50 to 52.
Conclusions
From 2019 to 2024, the average annual positive detection rate of influenza virus in Jiangsu Province remained relatively low. Influenza activity characterized by the alternating circulation of influenza A H1N1 subtype, influenza A H3N2 subtype, and influenza B Victoria linage. It is necessary to maintain the surveillance sensitivity for the influenza B Yamagata lineage.
3.Correlation between liver fibrosis degree and carotid plaque in patients with lean 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(2):319-325
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.
4.Construction of a biomimetic three-layered PLLA/PCL large-diameter vessel via electrospinning and ultrasonic pore-forming: Preliminary animal evaluation
Wenjun WANG ; Yang GAO ; Feng GAO ; Lei SHI ; Wei LIU ; Weiwang FAN ; Chang XU ; Hong ZHENG ; Xufeng DONG ; ZHUANG Xijing
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(07):1093-1100
Objective To fabricate a large-diameter vascular graft with a pore size gradient structure mimicking that of natural blood vessels, using poly-L-lactic acid (PLLA) and polycaprolactone (PCL) as base materials through electrospinning and ultrasonic pore-forming techniques, and to evaluate its application potential. Methods A three-layered tubular graft was fabricated from a PCL/PLLA blend (mass ratio 6 : 4) via electrospinning, followed by an ultrasonic pore-forming process to create a gradient porosity. The resulting graft (diameter: 2 cm, length: 4 cm) was implanted into the descending thoracic aorta of an experimental pig using an end-to-end anastomosis. Graft patency and anastomotic sites were monitored by computed tomography angiography (CTA) at 1 and 6 weeks post-surgery. After 2 months, the graft was explanted for systematic evaluation of vascular regeneration and repair through gross examination, histopathology (H&E and elastic fiber staining), immunohistochemistry [for ETS-related gene (ERG), Actin, and Vimentin], and scanning electron microscopy (SEM). Results Postoperative CTA confirmed excellent graft patency at both 1 and 6 weeks, with no evidence of thrombosis or anastomotic stenosis. Gross examination of the 2-month explant revealed a smooth luminal surface covered by neotissue. Histopathological analysis demonstrated that the graft successfully induced the formation of a three-layered structure resembling a native vessel wall, comprising endothelial cells, smooth muscle cells, and fibroblasts. Immunohistochemistry further verified coverage of the luminal surface by endothelial cells (ERG-positive), along with the presence of neosmooth muscle (Actin-positive) and fibroblasts (Vimentin-positive). Endothelial cells were observed adhering to the inner surface of the artificial vessel under SEM. Conclusion The biomimetic, three-layered PLLA/PCL large-diameter vascular graft, constructed via electrospinning and ultrasonic pore-forming, exhibits excellent short-term patency and biocompatibility in a large animal model. More importantly, it demonstrates a significant potential to promote host cell infiltration and achieve in situ regeneration of a three-layered vascular wall structure, providing a promising experimental basis for the development of next-generation functional vascular substitutes.
5.Comparative Study on the Effects of Qingchang Wenzhong Decoction (清肠温中方) on the Intestinal Mucus Barrier in Conventional and Pseudo-Germ-Free Model Mice of Acute Ulcerative Colitis
Mingxu GAO ; Leilei LIU ; Lijie LU ; Jiali WANG ; Juncong HU ; Yangzhe LIN ; Qirui LIU ; Yue DONG ; Luyue WANG ; Zeyu XUE ; Junxiang LI ; Lei SHI
Journal of Traditional Chinese Medicine 2026;67(14):1528-1537
ObjectiveTo explore the mechanisms of Qingchang Wenzhong Decoction (清肠温中方, QWD) in treating ulcerative colitis (UC). MethodsA total of 100 male C57BL/6J mice were assigned to a pseudo-germ-free group (n=45) and a conventional group (n=55) using a random number table. Pseudo-germ-free model was established by giving antibiotic treatment and subsequently subjected to acute UC induction. After the model was successfully established, the 45 pseudo-germ-free acute UC mice were randomly allocated to a pseudo-germ-free model group, a pseudo-germ-free QWD group, and a pseudo-germ-free mesalazine group, with 15 mice in each group. The 55 conventional mice were randomly divided into a normal control group (n=10), as well as a conventional model group, a conventional QWD group, and a conventional mesalazine group, with 15 mice in each group. Except for the normal control group, acute UC models were established in all other groups. Mice in the control group received no intervention. From day 32 of the experiment, mice in the pseudo-germ-free QWD group and the conventional QWD group were administered with QWD granules at a dose of 9.71 g/(kg·d) by gavage with deionized water, while those in the pseudo-germ-free mesalazine group and the conventional mesalazine group were given mesalazine enteric-coated tablets at a dose of 151.67 mg/(kg·d) by gavage. Mice in the remaining groups received an equal volume of deionized water once daily for 7 consecutive days. On day 25 of the experiment, six mice were randomly selected from both the pseudo-germ-free group and the conventional group to measure body weight, and fecal samples were collected for 16S rDNA sequencing to compare differences in gut microbiota, including operational taxonomic units (OTUs), Shannon index, Observed_species index, and phylogenetic diversity (PD whole tree). On day 39, body weight was measured in all groups, and the disease activity index (DAI) was evaluated. Colon length and colon wet weight were recorded. Histopathological changes in colonic tissues were assessed by hematoxylin and eosin (HE) staining, and histological injury scores were determined. In addition, the expression levels of mucopolysaccharides and mucin 2 (MUC2) in colonic tissues were measured. ResultsOn day 25, there was no significant difference in body weight between the pseudo-germ-free group and the conventional microbiota group (P>0.05). Compared to the conventional microbiota group, the pseudo-germ-free group showed significantly reduced OTUs, Shannon index, Observed_species index, and PD whole tree (P<0.01). On day 39, compared to the normal control group, mice in the conventional model group showed a significant decrease in the body weight and an increase in DAI, along with increased colon wet weight, shortened colon length, markedly elevated histopathological scores, and reduced expression of mucopolysaccharides and MUC2 (P<0.01). In contrast, no significant differences were observed in these parameters in the pseudo-germ-free model group (P>0.05). Compared to the conventional model group, the conventional QWD group showed significant improvement in all measured parameters, whereas the conventional mesalazine group showed reductions only in DAI scores and colon wet weight (P<0.01). No statistically significant differences were observed among the pseudo-germ-free model group, the pseudo-germ-free QWD group, and the pseudo-germ-free mesalazine group (P>0.05). ConclusionQWD may promote the reconstruction of the intestinal mucus barrier by regulating the intestinal flora, thereby exerting a therapeutic effect on acute UC.
6.Comparative Study on the Effects of Qingchang Wenzhong Decoction (清肠温中方) on the Intestinal Mucus Barrier in Conventional and Pseudo-Germ-Free Model Mice of Acute Ulcerative Colitis
Mingxu GAO ; Leilei LIU ; Lijie LU ; Jiali WANG ; Juncong HU ; Yangzhe LIN ; Qirui LIU ; Yue DONG ; Luyue WANG ; Zeyu XUE ; Junxiang LI ; Lei SHI
Journal of Traditional Chinese Medicine 2026;67(14):1528-1537
ObjectiveTo explore the mechanisms of Qingchang Wenzhong Decoction (清肠温中方, QWD) in treating ulcerative colitis (UC). MethodsA total of 100 male C57BL/6J mice were assigned to a pseudo-germ-free group (n=45) and a conventional group (n=55) using a random number table. Pseudo-germ-free model was established by giving antibiotic treatment and subsequently subjected to acute UC induction. After the model was successfully established, the 45 pseudo-germ-free acute UC mice were randomly allocated to a pseudo-germ-free model group, a pseudo-germ-free QWD group, and a pseudo-germ-free mesalazine group, with 15 mice in each group. The 55 conventional mice were randomly divided into a normal control group (n=10), as well as a conventional model group, a conventional QWD group, and a conventional mesalazine group, with 15 mice in each group. Except for the normal control group, acute UC models were established in all other groups. Mice in the control group received no intervention. From day 32 of the experiment, mice in the pseudo-germ-free QWD group and the conventional QWD group were administered with QWD granules at a dose of 9.71 g/(kg·d) by gavage with deionized water, while those in the pseudo-germ-free mesalazine group and the conventional mesalazine group were given mesalazine enteric-coated tablets at a dose of 151.67 mg/(kg·d) by gavage. Mice in the remaining groups received an equal volume of deionized water once daily for 7 consecutive days. On day 25 of the experiment, six mice were randomly selected from both the pseudo-germ-free group and the conventional group to measure body weight, and fecal samples were collected for 16S rDNA sequencing to compare differences in gut microbiota, including operational taxonomic units (OTUs), Shannon index, Observed_species index, and phylogenetic diversity (PD whole tree). On day 39, body weight was measured in all groups, and the disease activity index (DAI) was evaluated. Colon length and colon wet weight were recorded. Histopathological changes in colonic tissues were assessed by hematoxylin and eosin (HE) staining, and histological injury scores were determined. In addition, the expression levels of mucopolysaccharides and mucin 2 (MUC2) in colonic tissues were measured. ResultsOn day 25, there was no significant difference in body weight between the pseudo-germ-free group and the conventional microbiota group (P>0.05). Compared to the conventional microbiota group, the pseudo-germ-free group showed significantly reduced OTUs, Shannon index, Observed_species index, and PD whole tree (P<0.01). On day 39, compared to the normal control group, mice in the conventional model group showed a significant decrease in the body weight and an increase in DAI, along with increased colon wet weight, shortened colon length, markedly elevated histopathological scores, and reduced expression of mucopolysaccharides and MUC2 (P<0.01). In contrast, no significant differences were observed in these parameters in the pseudo-germ-free model group (P>0.05). Compared to the conventional model group, the conventional QWD group showed significant improvement in all measured parameters, whereas the conventional mesalazine group showed reductions only in DAI scores and colon wet weight (P<0.01). No statistically significant differences were observed among the pseudo-germ-free model group, the pseudo-germ-free QWD group, and the pseudo-germ-free mesalazine group (P>0.05). ConclusionQWD may promote the reconstruction of the intestinal mucus barrier by regulating the intestinal flora, thereby exerting a therapeutic effect on acute UC.
7.Pharmacoeconomic evaluation of brivaracetam versus perampanel as add-on therapy for drug-resistant focal epilepsy
Jiale QIAO ; Fanqi LIU ; Yuchen TANG ; Jie CHENG ; Yan SHI ; Jun QI ; Zhiqiang DONG
China Pharmacy 2026;37(18):2382-2388
OBJECTIVE To evaluate the cost-effectiveness of brivaracetam (BRV) versus perampanel (PER) as add-on therapy for drug-resistant focal epilepsy in adults from the perspective of the Chinese healthcare system.METHODS A Markov model was established to simulate the progression of drug-resistant focal epilepsy in adults, with a simulation cycle of 5 months, a time horizon of 44 years, and a discount rate of 4.5%. Clinical data required for the model were derived from the phase Ⅲ clinical trial of BRV and a post hoc analysis of PER; cost data and utility data were obtained from Yaozh and related literature. The willingness-to-pay (WTP) threshold was set at twice China’s 2025 per capita gross domestic product (GDP) [199 330 yuan per quality-adjusted life year (QALY)], and a cost-effectiveness analysis was adopted to compare the cost-effectiveness of the regimens. One-way sensitivity analysis and probabilistic sensitivity analysis were performed to confirm the stability of the base-case analysis conclusions.RESULTS The incremental cost-effectiveness ratio (ICER) of BRV 50 mg/d as an add-on therapy versus PER 8 mg/d was 44 823.08 yuan/QALY, which was below the WTP threshold; the ICER of BRV 200 mg/d versus PER 12 mg/d was 238 783.00 yuan/QALY, which was above the WTP threshold. In the one-way sensitivity analysis, the direct medical cost of seizures ≥53 times/year and the health utility value of a seizure-free state per year had a relatively large impact on the two ICER results mentioned above, respectively. Probabilistic sensitivity analysis indicated that the above results were robust.CONCLUSIONS For the selection of add-on therapy for drug-resistant focal epilepsy in adults in China, when twice China’s per capita GDP is used as the WTP threshold, BRV 50 mg/d has a cost-effectiveness advantage over PER 8 mg/d, whereas BRV 200 mg/d not have a cost-effectiveness advantage over PER 12 mg/d.
8.Assessment of annual effective dose for the public caused by the discharge of uranium-containing wastewater into river
Chang LIU ; Hailong CHEN ; Dong LIANG ; Linfeng SHI ; Hongwei CHAI
Chinese Journal of Radiological Health 2025;34(2):259-263
Objective To predict the radiation impact of discharging wastewater containing uranium within the specified limit generated during the normal operation of a new production line at a nuclear fuel plant on the receiving water body and its downstream, and to provide a reference for the management of radioactive liquid effluent discharge from nuclear facilities. Methods Based on the technical guidelines for environmental impact assessment, literature on radiation environmental impact assessment, and data collected from on-site investigations, appropriate hydrological parameters and prediction models were selected to analyze and predict the variation pattern of radioactive nuclide uranium along the receiving water body and the radiation exposure of nearby residents. Results The maximum increase in uranium concentration in the receiving water body and its downstream caused by the discharge of uranium-containing wastewater was 1.14 μg/L. The maximum predicted concentration was 2.75 μg/L after adding the background data of the water body. The resulting maximum individual annual effective dose for the public was 1.49 × 10−4 mSv/a. Conclusion The maximum predicted uranium concentration in the receiving water body and its downstream is lower than the uranium concentration limit of 30 μg/L specified in the Standards for Drinking Water Quality (GB 5749-2022). The maximum individual annual effective dose for the public is much lower than the control value of 0.2 mSv/a specified in the Radiation Protection Regulations for Uranium Processing and Fuel and Fuel Manufacturing Facilities (EJ 1056-2018). The radiation impact is acceptable.
9.Bibliographical cataloging for ancient TCM books
Hongtao LI ; Weina ZHANG ; Lin TONG ; Jingpeng DENG ; Qian ZHAO ; Honglei WANG ; Naiying LIU ; Mei SHI ; Qiang LIU ; Ying LIN ; Xiaohong ZHANG ; Lili FENG ; Mingrui ZHANG ; Yanqiu LUO ; Guangkun CHEN ; Yan DONG ; Bin LI ; Sihong LIU ; Bing LI ; Chen LI ; Meng LI ; Rui WANG ; He LU
International Journal of Traditional Chinese Medicine 2025;47(6):729-740
With reference to the Information and Documentation-Resource Description (GB/T 3792-2021) and Bibliographical Description for Ancient Chinese Books (GB/T 3792.7-2008) and other cataloging standards and rules, drawing on the practical experience of cataloging ancient TCM books, Bibliographical Cataloging for Ancient TCM Books was formulated. This standard specifies the entry items and their order of ancient TCM books, cataloging identifier, cataloging text, cataloging information source, and cataloging item details. The standard can provide standardized and unified guiding principles and methods for the work of ancient TCM books, and promote the sharing and utilization of ancient TCM books.
10.Effect and mechanism of UBAC2 mediated by m6A methylation modification on the invasion and migration abilities of colorectal cancer cells
Ying-Peng SHI ; Hua LIU ; Dong-Lin ZHANG ; Ya-Ping NI ; Jie CUI
Medical Journal of Chinese People's Liberation Army 2025;50(9):1162-1170
Objective To investigate the effects and mechanisms of ubiquitin-associated domain-containing protein 2(UBAC2)mediated by m6A methylation modification on the invasion and migration abilities of colorectal cancer cells.Methods The GEPIA2.0 database was utilized to analyze the expression differences of UBAC2 mRNA between colorectal cancer tissues and adjacent normal tissues,as well as its expression in colorectal cancer tissues at different stages.The correlation between Wilms tumor 1-associated protein(WTAP)and UBAC2 expression was analyzed.The Kaplan-Meier plotter online tool was applied to analyze the correlation between UBAC2 and the overall survival rate of colorectal cancer patients.The RMVar and SRAMP databases were employed to predict potential m6A methylation modification sites in the UBAC2 gene.Quantitative real-time PCR(qRT-PCR)and Western blotting were performed to detect the expression levels of UBAC2 mRNA and protein in colorectal cancer cell lines.For UBAC2 knockdown experiments,SW480 cells were divided into control group(no treatment),sh-NC group(transfected with sh-NC negative control plasmid),and sh-UBAC2 group(transfected with sh-UBAC2 plasmid).For WTAP knockdown experiments,groups included control group(no treatment),si-NC group(transfected with negative control siRNA),and si-WTAP group(transfected with WTAP-targeting siRNA).For UBAC2 overexpression experiments,groups were control group(no treatment),si-WTAP group(transfected with pcDNA3.1 empty plasmid),and si-WTAP+OE-UBAC2 group(transfected with UBAC2 overexpression plasmid pcDNA3.1-UBAC2).Western blotting was used to detect the protein expression levels of UBAC2,WTAP,E-cadherin,N-cadherin,and Vimentin;qRT-PCR was applied to detect the expression level of UBAC2 mRNA;Transwell assays were conducted to assess cell invasion and migration abilities.MeRIP-qPCR was employed to detect the m6A methylation modification of UBAC2 mRNA;RIP-qPCR experiments were conducted to verify the binding of WTAP to UBAC2 mRNA.In nude mouse colorectal cancer lung metastasis experiments,groups included LV-sh-NC group(tail vein injection of SW480 cells stably infected with LV-sh-NC)and LV-sh-UBAC2 group(tail vein injection of SW480 cells stably infected with LV-sh-UBAC2).After 42 d of tumor-implantation in nude mice,lung tissues were harvested:the number of lung nodules observed by hematoxylin/eosin(HE)staining,and the expression level of Luc2 mRNA detected by qRT-PCR.Results GEPIA2.0 database analysis revealed that the expression level of UBAC2 mRNA in colorectal cancer tissues was significantly higher than that in adjacent normal tissues,and it gradually increased with the progression of tumor stage(P<0.05).The expression levels of UBAC2 mRNA and protein in multiple colorectal cancer cell lines were significantly higher than those in normal colonic epithelial cells(P<0.05).Compared with sh-NC group,sh-UBAC2 group showed significantly increased E-cadherin protein expression,significantly decreased N-cadherin and Vimentin protein expression,and significantly reduced number of invaded and migrated SW480 cells(P<0.05).GEPIA2.0 database analysis results indicated a positive correlation between WTAP and UBAC2 expression(r=0.24,P<0.001).Compared with si-NC group,si-WTAP group showed significantly decreased expression levels of WTAP and UBAC2 mRNA and protein in SW480 cells(P<0.05).MeRIP-qPCR results demonstrated that the m6A modification level of UBAC2 mRNA in si-WTAP group was significantly lower than that in si-NC group(P<0.05).RIP-qPCR further confirmed that WTAP could bind to UBAC2 mRNA.Compared with control group,si-WTAP group showed significantly increased E-cadherin protein expression and significantly decreased N-cadherin and Vimentin protein expression in SW480 cells(P<0.05);compared with si-WTAP group,si-WTAP+OE-UBAC2 group showed significantly decreased E-cadherin protein expression and significantly increased N-cadherin and Vimentin protein expression in SW480 cells(P<0.05).The number of lung nodules in LV-sh-UBAC2 group was significantly fewer than that in LV-sh-NC group,and the expression level of Luc2 mRNA in lung tissues was significantly lower than that in LV-sh-NC group(P<0.05).Conclusion UBAC2 mediated by m6A methylation modification can regulate the epithelial-mesenchymal transition(EMT)process in colorectal cancer cells,thereby affecting the invasion and migration abilities of colorectal cancer cells.


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