Objective To explore the role of ring finger protein 41(RNF41)in the initiation and progression of cholangiocarcinoma.Methods The expression levels of RNF41 mRNA and protein in tumor tissues and adjacent normal tissues from 84 CHOL patients who underwent total surgical resection at the Second Affiliated Hospital of Kunming Medical University and Kunming Ganmei Hospital between January 2010 and December 2016 were analyzed using bioinformatics,Western blot,and immunohistochemistry.The TIMER 2.0 database was used to analyze the impact of RNF41 on the prognosis and survival of CHOL patients and the relationship between RNF41 and tumor clinical characteristics.RNF41 siRNA was transfected into HCC9810,RBE,and HUCCT1 cells.CCK-8,Edu,colony formation,and Western blot assays were conducted to evaluate the changes in proliferation of cholangiocarcinoma cells between the RNF41 knockdown group and the control group.Transwell assays and detection of EMT and migration markers were performed to assess changes in the invasion ability of cholangiocarcinoma cells between the RNF41 knockdown and control groups.Western blot was used to examine the effect of RNF41 knockdown on epithelial-mesenchymal transition in cholangiocarcinoma cells.Twelve BALB/c mice were randomly divided into two groups:a control group and an RNF41 knockdown group,with six mice in each group.Tumor formation assays,Western blot assays,and immunohistochemistry staining were carried out to investigate the effect of RNF41 knockdown on tumor growth in nude mice.Results Real-time quantitative fluorescence PCR analysis revealed that the expression level of RNF41 mRNA in cholangiocarcinoma tissues was significantly higher than that in the corresponding adjacent non-tumor tissues(P<0.01),and this trend was corroborated at the protein level by immunohistochemical staining.Using the TIMER 2.0 database,we further analyzed the correlation between RNF41 expression and clinicopathological features,including histological grade,tumor stage,lymph node metastasis,and patient survival.The results indicated that elevated RNF41 expression was significantly associated with advanced histological grade and lymph node metastasis in cholangiocarcinoma(P<0.01).Survival analysis demonstrated that high RNF41 expression was closely linked to poor prognosis in patients with cholangiocarcinoma(CHOL).Functional assays,including CCK-8,EdU incorporation,and colony formation,showed that RNF41 knockdown significantly inhibited the proliferation of cholangiocarcinoma cells compared with the control group.Western blot analysis revealed that,following RNF41 silencing,the expression of the epithelial-mesenchymal transition(EMT)marker E-cadherin was markedly upregulated,whereas the levels of mesenchymal markers N-cadherin and MMP9 were significantly reduced(P<0.05).These findings were consistent with the results obtained from in vitro experiments(P<0.01).Moreover,in vivo studies showed that RNF41 knockdown suppressed subcutaneous tumor formation in nude mice(P<0.05).Conclusion RNF41 plays a critical role in promoting the occurrence and progression of cholangiocarcinoma and is closely associated with adverse clinicopathological features and poor prognosis in patients.The knockdown of RNF41 effectively suppresses the proliferation,invasion,epithelial-mesenchymal transition(EMT),and tumorigenicity of cholangiocarcinoma cells.

Objective To study the correlation between the changes of matrix metalloproteinase-9(MMP-9)and neutrophil/lymphocyte ratio(NLR)before and after the revascularization of acute ischemic stroke(AIS),so as to find biomarkers to predict the bleeding transformation risk of AIS patients.Methods From February 2022 to December 2022,161 patients admitted to the Stroke Center of Qujing Hospital Affiliated to Kunming Medical University with AIS werre divided in to the hemorrhagic transformation group and the non-hemorrhagic transfor-mation groupand treated with revascularization(intravenous thrombolysis,endovascular treatment,combined the intravenous thrombolysis with endovascular treatment).Among them,there were 46 cases in the hemorrhagic transformation group and 115 cases in the non hemorrhagic transformation group.And the general data,NLR value and MMP-9 before and after the revascularization were compared between the two groups.Results There was no statistical difference in general data between the two groups(all P>0.05)except for C-reactive protein in hemorrhagic transformation group and in non-hemorrhagic transformation group(P<0.001).The white blood cells,neutrophils,neutrophil percentage,neutrophil absolute value,lymphocyte absolute value,NLR and MMP-9 value in hemorrhagic transformation group were significantly higher than those in non-hemorrhagic transformation group before the treatment and there was a statistical significance(P<0.05).After revascularization,the indexes of blood routine and MMP-9 were significantly higher than those before the revascularization,and the increase in hemorrhagic transformation group was more obvious than that in non-hemorrhagic transformation group and there was a statistical significance(P<0.001),The ROC curve showed that the area under the curve(AUC)of NLR and MMP-9 predicting bleeding transformation after AIS revascularization were 0.74 and 0.90.Conclusion NLR,MMP-9 are associated with the risk of bleeding transformation in AIS patients after the revascularization and can they can be used as the predictive factors for bleeding transformation risk.

Objective To establish an animal model of lung adenoma in BALB/c mice based on dynamic characterization by micro-computed tomography(CT).Methods Eighty female SPF-grade BALB/c mice were divided randomly into four groups:model low dose group(1 mg/g urethane,iP,once),model medium dose group(1 mg/g urethane,ip,once a week,followed by 2 weeks),model high dose group(1 mg/g urethane,ip,once a week,followed by 4 weeks),and blank group(equal volume of saline).Growth of lung nodules in the mice was monitored regularly using Micro-CT.Three-dimensional images of the lungs were drawn using the Analyze 12.0 system,and lung tissues were taken for histopathological examination(hematoxylin and eosin).Results Lung nodules with round high-density shadows were observed at week 11 in all model groups compared with the findings in the blank group.The rate of nodule formation increased with increasing modeling weeks,with rates of nodule formation in the model high,medium,and low dose groups of 93.8%,93.8%,and 87.5%,respectively,at week 21.Most mice had two to four,followed by one,and one to two nodules,respectively.The average maximum diameter of the lung nodules in the low dose group was significantly higher than the diameters in the medium-and high-dose groups(P＜0.05),but there was no significant difference in lung nodule volume among the three groups.Regarding pathological type,hematoxylin and eosin staining revealed that the tumors in all the model groups were lung adenomas.Conclusions Lung adenomas were successfully induced in all urethane dose groups of mice and growth of the lung nodules could be characterized by micro-CT.The rate of nodule formation was highest in the medium dose group,which developed a moderate number of lung adenomas and provided a stable model,and was thus considered the most suitable model for the study of lung adenomas in mice.

Acute mountain sickness(AMS),a condition characterized primarily by symptoms such as headache and nausea,has a high incidence and seriously affects the life and health of individuals undertaking rapid ascensions to high altitudes.The main pathophysiological manifestations of AMS include cerebral vasodilation and transient increases in intracranial pressure,with severe cases potentially incurring cerebral edema.The occurrence and development of AMS is associated with factors such as the susceptibility,physiological state,and psychological state of an individual.The molecular mechanisms involved include inflammatory responses,oxidative stress,immune regulation,and energy metabolism at the levels of genes,proteins,and metabolism.The management of AMS includes both prevention and treatment strategies.This article provides a comprehensive discussion of AMS from several aspects,including diagnosis,pathophysiological manifestations,susceptibility factors,molecular mechanisms,and prevention and treatment strategies.

Objective:To investigate the application of endothelial glycocalyx degradation products in assessing the severity of pulmonary edema in patients with acute respiratory distress syndrome (ARDS).Methods:A prospective study was conducted to select patients diagnosed with ARDS at Wuxi People's Hospital from July 1, 2018 to December 31, 2019. The extravascular lung water index (EVLWI) was recorded within 2 h after admission by continuous cardiac output with pulse indicator. The indexes of glycocalyx degradation products syndecan-1 (SDC-1), heparan sulfate (HS), hyaluronic acid (HA) and the concentrations of inflammatory factors [blood tumor necrosis factor α (TNF-α), interleukin (IL)-6 and IL-10] were measured by enzyme-linked immunosorbent assay. Pearson correlation method was adopted to analyze the correlation of glycocalyx degradation products with EVLWI and inflammatory factors in ARDS patients. The patients were divided into the mild pulmonary edema group and severe pulmonary edema group according to EVLWI at the cut-off value of 10 mL/kg, and the differences of glycocalyx degradation products and inflammatory factors between the two groups were compared. Receiver operating characteristic (ROC) curve of the subjects were plotted to analyze the value of glycocalyx degradation products in determining the severity of pulmonary edema.Results:A total of 85 ARDS patients were enrolled. Pearson correlation analysis showed that SDC-1, HS, and HA were all positively correlated with IL-6, TNF-α, EVLWI (all P<0.05), but did not correlate with IL-10 (all P>0.05). Comparison of indicators between the mild pulmonary edema group (39 cases) and the severe pulmonary edema group (46 cases) showed that: IL-6[(33.63±3.43) ng/L vs. (39.99±4.64) ng/L], TNF-α[(43.38±6.05) ng/L vs. (50.79±7.35) ng/L], SDC-1[(494.13±47.23) ng/L vs. (563.50±56.36) ng/L], HS[(114.02±18.39) ng/mL vs. (138.93±17.02) ng/mL], and HA[(441.44±62.52) ng/mL vs. (546.23±85.24) ng/mL] were statistically different between the two groups(all P<0.05). Whereas, IL-10 [(24.37±10.11) ng/L vs. (28.75±11.98) ng/L] was not statistically different between the two groups ( P>0.05). ROC curve analysis showed that the combined prediction of SDC-1, HA and HS indicators was superior to the single indicator. The area under the ROC curve combining the three indicators was 0.928 (95% CI: 0.872-1.000), with a sensitivity and specificity of 87.5% and 86.7%, respectively. Conclusions:There is a positive correlation between glycocalyx degradation products SDC-1, HS, HA and EVLWI in ARDS patients. The application of these three glycocalyx degradation products can be used as a reliable indicators for judging the severity of pulmonary edema in ARDS patients.

Mevalonate metabolism plays an important role in regulating tumor growth and progression; however, its role in immune evasion and immune checkpoint modulation remains unclear. Here, we found that non-small cell lung cancer (NSCLC) patients with higher plasma mevalonate response better to anti-PD-(L)1 therapy, as indicated by prolonged progression-free survival and overall survival. Plasma mevalonate levels were positively correlated with programmed death ligand-1 (PD-L1) expression in tumor tissues. In NSCLC cell lines and patient-derived cells, supplementation of mevalonate significantly up-regulated the expression of PD-L1, whereas deprivation of mevalonate reduced PD-L1 expression. Mevalonate increased CD274 mRNA level but did not affect CD274 transcription. Further, we confirmed that mevalonate improved CD274 mRNA stability. Mevalonate promoted the affinity of the AU-rich element-binding protein HuR to the 3'-UTR regions of CD274 mRNA and thereby stabilized CD274 mRNA. By in vivo study, we further confirmed that mevalonate addition enhanced the anti-tumor effect of anti-PD-L1, increased the infiltration of CD8⁺ T cells, and improved cytotoxic function of T cells. Collectively, our findings discovered plasma mevalonate levels positively correlated with the therapeutic efficacy of anti-PD-(L)1 antibody, and provided the evidence that mevalonate supplementation could be an immunosensitizer in NSCLC.

Females increase aggression for mating opportunities and for acquiring reproductive resources. Although the close relationship between female aggression and mating status is widely appreciated, whether and how female aggression is regulated by mating-related cues remains poorly understood. Here we report an interesting observation that Drosophila virgin females initiate high-frequency attacks toward mated females. We identify 11-cis-vaccenyl acetate (cVA), a male-derived pheromone transferred to females during mating, which promotes virgin female aggression. We subsequently reveal a cVA-responsive neural circuit consisting of four orders of neurons, including Or67d, DA1, aSP-g, and pC1 neurons, that mediate cVA-induced virgin female aggression. We also determine that aSP-g neurons release acetylcholine (ACh) to excite pC1 neurons via the nicotinic ACh receptor nAChRα7. Together, beyond revealing cVA as a mating-related inducer of virgin female aggression, our results identify a neural circuit linking the chemosensory perception of mating-related cues to aggressive behavior in Drosophila females.
Animals ; Male ; Female ; Drosophila/physiology* ; Drosophila Proteins/physiology* ; Cues ; Sexual Behavior, Animal/physiology* ; Aggression/physiology* ; Drosophila melanogaster/physiology*

Abnormally activated CDK9 participates in the super-enhancer mediated transcription of short-lived proteins required for cancer cell survival. Targeting CDK9 has shown potent anti-tumor activity in clinical trials among different cancers. However, the study and knowledge on drug resistance to CDK9 inhibitors are very limited. In this study, we established an AML cell line with acquired resistance to a highly selective CDK9 inhibitor BAY1251152. Through genomic sequencing, we identified in the kinase domain of CDK9 a mutation L156F, which is also a coding SNP in the CDK9 gene. By knocking in L156F into cancer cells using CRISPR/Cas9, we found that single CDK9 L156F could drive the resistance to CDK9 inhibitors, not only ATP competitive inhibitor but also PROTAC degrader. Mechanistically, CDK9 L156F disrupts the binding with inhibitors due to steric hindrance, further, the mutation affects the thermal stability and catalytic activity of CDK9 protein. To overcome the drug resistance mediated by the CDK9-L156F mutation, we discovered a compound, IHMT-CDK9-36 which showed potent inhibition activity both for CDK9 WT and L156F mutant. Together, we report a novel resistance mechanism for CDK9 inhibitors and provide a novel chemical scaffold for the future development of CDK9 inhibitors.

[This corrects the article DOI: 10.1016/j.apsb.2023.04.002.].

During the carcinoma transformation of colitis， the imbalance of “metabolic-immune interaction” resulted from abnormal energy and metabolic substrates flow and direction was the key process， which caused by intercellular metabolic competition. Based on traditional Chinese medicine （TCM） theory and clinical research， we found that “fire failing to warm earth” is the key pathogenesis of colon-cancer transformation. “Fire” was a synonym for TCM to understand the energy metabolism， combined modern medical research findings， we thought energy metabolism disorders was a microcosmic manifestation of the “fire decline”， while abnormal immune function was the biological basis of “earth deficiency”. The imbalance between “metabolism-immune interaction” and the “fire failing to warm earth” pathogenesis of colitis-cancer transformation demonstrated the different understanding of the same pathological mechanism between western medicine and TCM. For treatment， it could be effectivce to delay the transformation of colitis-cancer by synergistically regulated the energy metabolism - “replenish fire” and enhanced the immune function - “nourish earth”， which was called the methods of replenishing fire to nourish earth.