ObjectiveHepatocellular carcinoma (HCC) represents 90% of all primary liver cancers. The main risk factors associated with HCC include viral hepatitis (B and/or C), alcohol abuse, and metabolic dysfunction-associated steatotic liver disease (MASLD), which progressively advance to liver fibrosis, cirrhosis, and ultimately evolve into HCC. Surgical resection represents the most effective treatment for HCC, while recent advances in immunotherapy, including immune checkpoint inhibitors and adoptive cell therapies, have provided improved treatment prospects for patients with unresectable HCC. However, the complex metabolic heterogeneity of HCC limits the therapeutic efficacy. Metabolic intermediates acyl-CoA not only provide energy and substrates for numerous biochemical reactions but also serve as donors for protein lysine acylation, a major class of post-translational modification (PTM). Therefore, a deeper understanding of the molecular mechanisms underlying protein lysine acylation and hepatocarcinogenesis is urgently needed. MethodsThe levels of protein lysine acylation and silence information regulator 5 (SIRT5) expression levels in clinical HCC samples were analyzed by Western blot. Quantitative malonylome and succinylome of HCC samples were analyzed by antibody-based affinity enrichment coupled with tandem mass spectrometry. The proliferation of HCC cells was analyzed with Cell Counting Kit-8 (CCK-8) assays, the apoptosis was quantified by Annexin V-FITC/propidium iodide (PI) staining coupled with flow cytometry, and the ability of cells to migrate was assayed by Transwell assays. The enzymatic activity of glutathione S-transferase Mu 1 (GSTM1) was quantified. Transgenic mice with hepatic overexpression of SIRT5 were constructed using CRISPR-Cas9, and primary hepatocarcinogenesis was induced by administration of diethylnitrosamine. ResultsWestern blot analysis indicated that the expression level of SIRT5 was elevated in clinical samples from HCC patients, and the levels of lysine malonylation, glutarylation, and succinylation were significantly reduced in HCC tissues. Knockout of SIRT5 in MHCC-97H and MHCC-97L hepatoma cells suppressed cell proliferation, and increased the percentage of apoptotic cells significantly. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of the differentially malonylome and succinylome of HCC samples revealed significant enrichment in two major classes of biological processes: core energy metabolism (e.g., glycolysis/gluconeogenesis, tricarboxylic acid metabolic process, fatty acid beta oxidation) and detoxification and oxidative stress response (e.g., response to toxic substance, chemical carcinogenesis, reactive oxygen species (ROS)). SIRT5 removes malonylation from lysine residues in GSTM1 and restores its detoxification activity, which is crucial for the survival of hepatocytes under stressed conditions. More importantly, in vivo experiment indicated that hepatic-specific overexpression of SIRT5 in mice accelerated diethylnitrosamine-induced liver fibrosis and hepatocarcinogenesis, indicating the critical role of SIRT5 in HCC progression. ConclusionThis study highlights the previously unrecognized SIRT5-GSTM1 axis as a key regulator in hepatocarcinogenesis, and suggests a potential target for the treatment of patients with HCC.

Objective To investigate the effect of long non-coding RNA(lncRNA)potassium voltage-gated channel subfamily Q member 1 overlapping transcript 1(KCNQ1OT1)on oxidative glucose deprivation/reoxygenation(OGD/R)-induced microglia injury through regulating the miR-145-5p/Rho-associated coiled-coil forming protein kinase 1(ROCK1)axis.Methods Microglia N9 were divided into the control group(normal culture),the OGD/R group(OGD/R-induced injury),the sh-NC group(transfected with sh-NC after OGD/R-induced injury),the sh-KCNQ1OT1 group(transfected with sh-KCNQ1OT1 after OGD/R-induced injury),the sh-KCNQ1OT1+inhibitor NC group(co-transfected with sh-KCNQ1OT1 and inhibitor NC after OGD/R-induced injury),and the sh-KCNQ1OT1+miR-145-5p inhibitor group(co-transfected with sh-KCNQ1OT1 and miR-145-5p inhibitor after OGD/R-induced injury).RT-qPCR was applied to detect the expression of lncRNA KCNQ1OT1,miR-145-5p,and ROCK1 mRNA in cells.The expression of ROCK1 protein was detected by Western blot.CCK-8 was applied to detect the cell proliferation.Flow cytometry was applied to detect cell apoptosis.ELISA was applied to detect the levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)in cells.The action sites of miR-145-5p with lncRNA KCNQ1OT1 and ROCK1 were predicted by bioinformatics website,and their targeting relationships were verified by dual-luciferase reporter assay.Results Compared with the control group,the expression of lncRNA KCNQ1OT1,the expression of ROCK1 mRNA and protein,the apoptosis rate,and the levels of IL-6,TNF-α,and IL-1β in cells were all increased,and the level of miR-145-5p and cell survival rate were all decreased in the OGD/R group(P<0.05).Compared with the OGD/R group and the sh-NC group,the expression of lncRNA KCNQ1OT1,the expression of ROCK1 mRNA and protein,the apoptosis rate,and the levels of IL-6,TNF-α,and IL-1β in cells were all decreased,and the level of miR-145-5p and cell survival rate were all increased in the sh-KCNQ1OT1 group,with significant differences(P<0.05).Compared with the sh-KCNQ1OT1+inhibitor NC group,the expression of ROCK1 mRNA and protein,the apoptosis rate,and the levels of IL-6,TNF-α,and IL-1β in cells were all increased,and the expression of miR-145-5p and cell survival rate were all decreased in the sh-KCNQ1OT1+miR-145-5p inhibitor group,with significant differences(P<0.05).Bioinformatics website showed that miR-145-5p had targeted action sites with lncRNA KCNQ1OT1 and ROCK1,and dual-luciferase reporter assay confirmed that miR-145-5p had targeting relationships with lncRNA KCNQ1OT1 and ROCK1(P<0.05).Conclusion Silencing lncRNA KCNQ1OT1 can alleviate OGD/R-induced microglia injury via upregulating the expression of miR-145-5p and targeting the down-regulation of ROCK1 expression.

Objective To obtain measurements of fetal four-chamber view and left and right ventricular shapes using two-dimensional speckle tracking,and to explore the clinical quantification of cardiac shape remodeling in small for gestational age(SGA)fetuses.Methods In this study,we prospectively collected data on singleton pregnancies from 28 to 39 weeks that were established in the archives of Obstetrics and Gynecology Hospital,Fudan University from May 2020 to Jul 2021.Fetuses eligible for inclusion criteria were randomly matched according to the ratio of estimated fetal weight(EFW)≥10th percentile(P10)∶EFWP90 in the apical segments of the left ventricle was greater in normal UAPI fetuses than that in low-risk fetuses(24.2%vs.14.3%,P=0.005);the SI of left ventricle in UAPI-normal SGA fetuses was greater in segments S1 to S22 than that of right ventricle with statistically significant difference(all P<0.05),while no significant difference was found between the SI of left and right ventricles in segments S23 and S24.Conclusion About one-fourth of SGA fetuses with normal UAPI have abnormally increased apical segments of the left ventricular SI,suggesting that cardiac remodeling with a predominantly"elongated"apical segments of the left ventricle precedes Doppler anomalies.

Objective To investigate the clinical characteristics of preoperative intestinal symptoms in patients with bowel endometriosis and to compare the effects of shaving versus segmental bowel resection on postoperative intestinal function.Methods A total of 105 patients diagnosed with bowel endometriosis and treated by the same surgical team at the Obstetrics and Gynecology Hospital,Fudan University between Aug 1,2013 and Dec 30,2017 were prospectively enrolled in this study.Clinical data were collected via outpatient visits and telephone follow-ups at four time points:preoperative(T0)and postoperative(T1:Nov 2018;T2:Nov 2020;T3:Apr 2024).The primary outcome was bowel symptoms and gastrointestinal function scores;secondary outcome was pain scores.A generalized estimating equation(GEE)model was used to analyze the interaction effect of surgical approach and follow-up time on outcomes.Results Ultimately,a total of 89 patients were included(15.24%loss to follow-up),among whom 79 patients(88.76%)underwent shaving excision.Preoperatively,46 patients(51.68%)presented with bowel symptoms,primarily anus bulge(21 cases,46.65%)and diarrhea(15 cases,32.61%)during menstruation.Postoperatively,there was a significant increase in constipation rates(T1:71.43%;T2:50.00%;T3:72.00%).Both surgical groups exhibited significant improvements in dysmenorrhea,gastrointestinal discomfort scores as well as gastrointestinal quality of life index(P<0.000 5).However,the segmental resection group had significantly higher scores for low anterior resection syndrome,constipation compared with the shaving excision group(P=0.02 and P=0.05).Conclusion Approximately half of the patients with bowel endometriosis exhibit typical bowel symptoms preoperatively,such as anus bulge and diarrhea.Both shaving excision and segmental resection effectively alleviate pain;however,shaving excision demonstrates an advantage regarding preservation of bowel function,whereas segmental resection may elevate risks associated with postoperative constipation or altered defecation patterns due to structural changes.The selection of surgical approach should carefully balance lesion removal and functional preservation,moreover,be sure that potential risks are thoroughly informed to patients prior to surgery.

Objective To obtain measurements of fetal four-chamber view and left and right ventricular shapes using two-dimensional speckle tracking,and to explore the clinical quantification of cardiac shape remodeling in small for gestational age(SGA)fetuses.Methods In this study,we prospectively collected data on singleton pregnancies from 28 to 39 weeks that were established in the archives of Obstetrics and Gynecology Hospital,Fudan University from May 2020 to Jul 2021.Fetuses eligible for inclusion criteria were randomly matched according to the ratio of estimated fetal weight(EFW)≥10th percentile(P10)∶EFWP90 in the apical segments of the left ventricle was greater in normal UAPI fetuses than that in low-risk fetuses(24.2%vs.14.3%,P=0.005);the SI of left ventricle in UAPI-normal SGA fetuses was greater in segments S1 to S22 than that of right ventricle with statistically significant difference(all P<0.05),while no significant difference was found between the SI of left and right ventricles in segments S23 and S24.Conclusion About one-fourth of SGA fetuses with normal UAPI have abnormally increased apical segments of the left ventricular SI,suggesting that cardiac remodeling with a predominantly"elongated"apical segments of the left ventricle precedes Doppler anomalies.

Objective To investigate the clinical characteristics of preoperative intestinal symptoms in patients with bowel endometriosis and to compare the effects of shaving versus segmental bowel resection on postoperative intestinal function.Methods A total of 105 patients diagnosed with bowel endometriosis and treated by the same surgical team at the Obstetrics and Gynecology Hospital,Fudan University between Aug 1,2013 and Dec 30,2017 were prospectively enrolled in this study.Clinical data were collected via outpatient visits and telephone follow-ups at four time points:preoperative(T0)and postoperative(T1:Nov 2018;T2:Nov 2020;T3:Apr 2024).The primary outcome was bowel symptoms and gastrointestinal function scores;secondary outcome was pain scores.A generalized estimating equation(GEE)model was used to analyze the interaction effect of surgical approach and follow-up time on outcomes.Results Ultimately,a total of 89 patients were included(15.24%loss to follow-up),among whom 79 patients(88.76%)underwent shaving excision.Preoperatively,46 patients(51.68%)presented with bowel symptoms,primarily anus bulge(21 cases,46.65%)and diarrhea(15 cases,32.61%)during menstruation.Postoperatively,there was a significant increase in constipation rates(T1:71.43%;T2:50.00%;T3:72.00%).Both surgical groups exhibited significant improvements in dysmenorrhea,gastrointestinal discomfort scores as well as gastrointestinal quality of life index(P<0.000 5).However,the segmental resection group had significantly higher scores for low anterior resection syndrome,constipation compared with the shaving excision group(P=0.02 and P=0.05).Conclusion Approximately half of the patients with bowel endometriosis exhibit typical bowel symptoms preoperatively,such as anus bulge and diarrhea.Both shaving excision and segmental resection effectively alleviate pain;however,shaving excision demonstrates an advantage regarding preservation of bowel function,whereas segmental resection may elevate risks associated with postoperative constipation or altered defecation patterns due to structural changes.The selection of surgical approach should carefully balance lesion removal and functional preservation,moreover,be sure that potential risks are thoroughly informed to patients prior to surgery.

Objective To explore the efficacy of single and combined parameters from 4 non-Gaussian diffusion models for differentiating breast imaging reporting and data system(BI-RADS)MRI category 4 benign and malignant breast tumors.Methods A total of 161 BI-RADS MRI category 4 breast lesions from 159 patients were retrospectively enrolled.Based on pathological results,the lesions were divided into malignant group(n=132)and benign group(n=29).The apparent diffusion coefficient(ADC)values were calculated from diffusion weighted imaging(DWI)sequences.Multi-b-value diffusion imaging data were acquired and fitted using 4 non-Gaussian models to obtain respective parameters,including diffusion kurtosis imaging(DKI),stretched exponential model(SEM),continuous-time random walk(CTRW)and fractional order calculus(FROC)model.Univariable and multivariable logistic regression analyses were employed to identify the diffusion quantitative indicators useful for differentiating benign and malignant BI-RADS MRI category 4 breast tumors.Receiver operating characteristic(ROC)curves were drawn,and the optimal threshold was determined using Youden index.The differentiating performance of ADC value,single parameter and their combination from non-Gaussian diffusion models were assessed and compared according to the area under the curve(AUC)of ROC curves,as well as the sensitivity,specificity and accuracy under the optimal thresholds.Results In malignant group,αCTRW,KDKI and μFROC values were higher,while ADC,DCTRW,DFROC,DDCSEM,αSEM and DDKI values were lower than those in benign group(all P＜0.05).Multivariable logistic regression analysis identified DCTRW and αCTRW values as independent factors for differentiating benign and malignant BI-RADS MRI category 4 breast tumors(both P＜0.05),and a combined model was then constructed.The AUC of the combined model was higher than that of each single parameter including ADC,DCTRW and αCTRW values(all P＜0.05).Conclusion The combined model of DCTRW and αCTRW had better efficacy than each single parameter for differentiating benign and malignant BI-RADS MRI category 4 breast tumors.

Objective To compare the distribution characteristics and drug resistance of mucous and non-mucous Streptococcus pneumoniae(SP).Methods SP isolated from clinical specimens of the Second Affiliated Hospital of Xiamen Medical College from January 2023 to June 2024 were collected.They were isolated,cultured and identified,and their susceptibility to antibacterial drugs was determined.Results A total of 291 SP strains were isolated,of which 20 strains were mucous SP and 271 strains were non-mucous SP.Children played a dominant role of non-mucous SP infection,but mucous SP mainly infected adults and the children above 5 years old.Both phenotypes of SP were mainly characterized by pulmonary infection.Non-mucous SP had a relatively high sensitivity rate to vancomycin,linezolid,penicillin,ertapenem,chloramphenicol,levofloxacin,moxifloxacin and ofloxacin.It had a high resistance rate to compound sulfamethoxazole,clindamycin,erythromycin and tetracycline.Mucous SP was completely sensitive to penicillin,amoxicillin,cefotaxime,ceftriaxone,ertapenem,vancomycin,linezolid,levofloxacin and moxifloxacin,and had a relatively high resistance rate to clindamycin,erythromycin and tetracycline.Conclusion The laboratory should enhance the detection capacity of mucous SP.Mucous SP is highly sensitive to common antibiotics.Clinicians should select antibiotics based on drug sensitivity results to delay the occurrence of drug resistance.

The prelimbic cortex (PL) plays a critical role in processing both the sensory and affective components of pain. However, the underlying molecular mechanisms remain poorly understood. In this study, we observed a reduction in hyperpolarization-activated cation current (I_h) in layer V pyramidal neurons of the contralateral PL in a mouse model of spared nerve injury (SNI). The expression of hyperpolarization-activated cyclic nucleotide-gated 2 (HCN2) channels was also decreased in the contralateral PL. Conversely, microinjection of fisetin, a partial agonist of HCN2, produced both analgesic and anxiolytic effects. Additionally, we found that cyclin-dependent kinase 5 (CDK5) was activated in the contralateral PL, where it formed a complex with HCN2 and phosphorylated its C-terminus. Knockdown of CDK5 restored HCN2 expression and alleviated both pain hypersensitivity and anxiety-like behaviors. Collectively, these results indicate that CDK5-mediated dysfunction of HCN2 in the PL underlies nerve injury-induced mechanical hypersensitivity and anxiety.
Animals ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels/metabolism* ; Hyperalgesia/metabolism* ; Cyclin-Dependent Kinase 5/metabolism* ; Neuralgia/metabolism* ; Male ; Anxiety/metabolism* ; Mice ; Potassium Channels/metabolism* ; Mice, Inbred C57BL ; Disease Models, Animal ; Pyramidal Cells/metabolism*