Objective:To investigate the immune signatures of endometrial carcinoma(EC)patients with POLE mutation.Methods:A total of 104 paraffin-embedded tissue samples of EC were collected from December 2020 to October 2022,including 11 POLE mutants and 93 POLE wild type in Department of Pathology,Taihe Hospital.Immunohistochemistry was conducted to detect ex-pressions of CD4,CD8 and PD-L1 proteins in EC tissue.Combined with TCGA database,bioinformatics was used to analyze immune infiltration of POLE mutants,mainly including CD4,CD8,PD-L1 mRNA expressions and related prognosis analysis.Results:Based on TCGA database analysis,compared to POLE wild type EC,significantly different expression genes,including chemokine family,immune cell surface markers and lysozyme genes(all P<0.05),were revealed in POLE mutants.In POLE mutant samples,main enriched signaling pathways including immune regulation,mesenchymal-epithelial transformation,etc.(all P<0.01).Compared with POLE wild-type,abundance of CD4+T cells,CD8+T cells,M1 macrophages and dendritic cells in EC samples with POLE mutation was sig-nificantly up-regulated(P<0.05).TISIDB platform assay suggested that POLE mutants primarily drove biological behavior of immune cells,including recruitment and activation of dendritic cells,NK cells,T cells and other immune cells,and lymphocyte homing in EC.Additionally,various immunosuppressive genes(including PD-L1,CTLA-4,TIM-3,etc.)were significantly overexpressed in EC with POLE mutants.In clinical samples,it was also confirmed that POLE mutation was significantly correlated with hyper-expressions of CD8,CD4 and PD-L1(all P<0.001).Prognostic analysis of TCGA showed that survival time of CD8 or PD-L1 over-expression of the POLE mutant subgroup was significantly prolonged,while CD8 or PD-L1 down-regulation of POLE wild-type subgroup was the shortest(P<0.01);there was no significant difference in CD4 subgroup(P>0.05).Conclusion:In EC patients,infiltration abun-dance of CD8+T cells and expression of PD-L1 have crucial value for prognosis or treatment of EC patients with POLE mutation.

Objective:To investigate the application value of multi-parameter ultrasound in the non-invasive assessment of renal fibrosis in patients with chronic kidney disease（CKD）.Methods:From December 2023 to April 2024，77 CKD patients（CKD group）and 30 healthy individuals（control group）from Renmin Hospital of Wuhan University were prospectively collected. The CKD group was further classified into mild（ n = 30），moderate（ n = 25）and severe（ n = 22）groups according to pathological assessment of renal fibrosis. Ultrasound parameters and clinical data of all patients were collected for group comparisons. Ultrasound parameters that showed statistically significant differences in univariate analysis were incorporated into a Logistic regression model to identify independent influencing factors. ROC curve was constructed，and the area under the curve（AUC）was calculated for statistical evaluation. Results:① The moderate and severe groups compared to control group，as well as the severe group compared to the mild and moderate groups，showed increased levels of SCr and BUN，and decreased level of eGFR，with statistically significant differences（all P < 0.05）. The kidney length and renal parenchymal thickness in the moderate group were smaller than those in the control group，while the kidney length，renal parenchymal thickness，and renal cortical thickness in the severe group were smaller than those in the control，mild，and moderate groups，with statistically significant differences among the groups（all P < 0.05）. The Vmax in the severe group was lower than those in the control，mild，and moderate groups，while the RI was higher than those in the control，mild，and moderate groups，with statistically significant differences（all P < 0.05）. ②In superb microvascular imaging，perfusion levels were predominantly grade 5 in the control and mild groups（83.3%，25/30；70.0%，21/30），grade 4 in the moderate group（48.0%，12/25），and grade 3 in the severe group（63.6%，14/22）. No significant differences were observed between the mild and control groups，or between the moderate and severe groups（all P>0.05），significant differences in blood flow perfusion were found between other groups（ P < 0.05）.③Shear wave elastography and super-resolution imaging revealed that mean of Young's modulus（Emean）in renal cortex increased progressively from the mild to the severe group，while microvascular density decreased. Except for Emean，which did not show significant differences between the moderate and severe groups（ P > 0.05），all other parameters showed significant differences between groups（ P < 0.05）. ④ROC curve analysis indicated that differentiating the control group from the mild group using Emean and microvascular density，the AUC was 0.769（95% CI = 0.631 - 0.872， P<0.001），with a sensitivity of 0.821 and specificity of 0.539. For the combined use of Emean，microvascular density，peak inter-arterial velocity，and resistance index for distinguishing mild from moderate-to-severe fibrosis resulted in an AUC of 0.902（95% CI = 0.843 - 0.972，P < 0.001），with a sensitivity of 0.967 and specificity of 0.755. These multi-parameter combined diagnostic approaches outperformed single ultrasound parameters. Conclusions:Multi-parameter ultrasound technology can display multidimensional renal changes in patients with CKD and effectively assess the degree of renal fibrosis. It is expected to become a non-invasive，highly sensitive tool for evaluating renal fibrosis in CKD.

Objective:To investigate the immune signatures of endometrial carcinoma(EC)patients with POLE mutation.Methods:A total of 104 paraffin-embedded tissue samples of EC were collected from December 2020 to October 2022,including 11 POLE mutants and 93 POLE wild type in Department of Pathology,Taihe Hospital.Immunohistochemistry was conducted to detect ex-pressions of CD4,CD8 and PD-L1 proteins in EC tissue.Combined with TCGA database,bioinformatics was used to analyze immune infiltration of POLE mutants,mainly including CD4,CD8,PD-L1 mRNA expressions and related prognosis analysis.Results:Based on TCGA database analysis,compared to POLE wild type EC,significantly different expression genes,including chemokine family,immune cell surface markers and lysozyme genes(all P<0.05),were revealed in POLE mutants.In POLE mutant samples,main enriched signaling pathways including immune regulation,mesenchymal-epithelial transformation,etc.(all P<0.01).Compared with POLE wild-type,abundance of CD4+T cells,CD8+T cells,M1 macrophages and dendritic cells in EC samples with POLE mutation was sig-nificantly up-regulated(P<0.05).TISIDB platform assay suggested that POLE mutants primarily drove biological behavior of immune cells,including recruitment and activation of dendritic cells,NK cells,T cells and other immune cells,and lymphocyte homing in EC.Additionally,various immunosuppressive genes(including PD-L1,CTLA-4,TIM-3,etc.)were significantly overexpressed in EC with POLE mutants.In clinical samples,it was also confirmed that POLE mutation was significantly correlated with hyper-expressions of CD8,CD4 and PD-L1(all P<0.001).Prognostic analysis of TCGA showed that survival time of CD8 or PD-L1 over-expression of the POLE mutant subgroup was significantly prolonged,while CD8 or PD-L1 down-regulation of POLE wild-type subgroup was the shortest(P<0.01);there was no significant difference in CD4 subgroup(P>0.05).Conclusion:In EC patients,infiltration abun-dance of CD8+T cells and expression of PD-L1 have crucial value for prognosis or treatment of EC patients with POLE mutation.

Objective:To investigate the application value of multi-parameter ultrasound in the non-invasive assessment of renal fibrosis in patients with chronic kidney disease（CKD）.Methods:From December 2023 to April 2024，77 CKD patients（CKD group）and 30 healthy individuals（control group）from Renmin Hospital of Wuhan University were prospectively collected. The CKD group was further classified into mild（ n = 30），moderate（ n = 25）and severe（ n = 22）groups according to pathological assessment of renal fibrosis. Ultrasound parameters and clinical data of all patients were collected for group comparisons. Ultrasound parameters that showed statistically significant differences in univariate analysis were incorporated into a Logistic regression model to identify independent influencing factors. ROC curve was constructed，and the area under the curve（AUC）was calculated for statistical evaluation. Results:① The moderate and severe groups compared to control group，as well as the severe group compared to the mild and moderate groups，showed increased levels of SCr and BUN，and decreased level of eGFR，with statistically significant differences（all P < 0.05）. The kidney length and renal parenchymal thickness in the moderate group were smaller than those in the control group，while the kidney length，renal parenchymal thickness，and renal cortical thickness in the severe group were smaller than those in the control，mild，and moderate groups，with statistically significant differences among the groups（all P < 0.05）. The Vmax in the severe group was lower than those in the control，mild，and moderate groups，while the RI was higher than those in the control，mild，and moderate groups，with statistically significant differences（all P < 0.05）. ②In superb microvascular imaging，perfusion levels were predominantly grade 5 in the control and mild groups（83.3%，25/30；70.0%，21/30），grade 4 in the moderate group（48.0%，12/25），and grade 3 in the severe group（63.6%，14/22）. No significant differences were observed between the mild and control groups，or between the moderate and severe groups（all P>0.05），significant differences in blood flow perfusion were found between other groups（ P < 0.05）.③Shear wave elastography and super-resolution imaging revealed that mean of Young's modulus（Emean）in renal cortex increased progressively from the mild to the severe group，while microvascular density decreased. Except for Emean，which did not show significant differences between the moderate and severe groups（ P > 0.05），all other parameters showed significant differences between groups（ P < 0.05）. ④ROC curve analysis indicated that differentiating the control group from the mild group using Emean and microvascular density，the AUC was 0.769（95% CI = 0.631 - 0.872， P<0.001），with a sensitivity of 0.821 and specificity of 0.539. For the combined use of Emean，microvascular density，peak inter-arterial velocity，and resistance index for distinguishing mild from moderate-to-severe fibrosis resulted in an AUC of 0.902（95% CI = 0.843 - 0.972，P < 0.001），with a sensitivity of 0.967 and specificity of 0.755. These multi-parameter combined diagnostic approaches outperformed single ultrasound parameters. Conclusions:Multi-parameter ultrasound technology can display multidimensional renal changes in patients with CKD and effectively assess the degree of renal fibrosis. It is expected to become a non-invasive，highly sensitive tool for evaluating renal fibrosis in CKD.

A 36-year-old male patient was hospitalized due to persistent chest tightness and dyspnea last-ing over 10 days.An echocardiogram revealed significant aortic valve regurgitation,the presence of vegetation on the aortic valve,severe mitral valve regurgitation,and enlargement of both the left atrium and left ventricle.Follow-ing a comprehensive multidisciplinary consultation,the focus was directed towards the aortic valve vegetation as the primary concern,with particular attention given to the potential diagnosis of blood culture-negative infective endocarditis(IE)and non-bacterial thrombotic endocarditis,while differentiating it from rheumatic heart disease.Ultimately,the vegetation was excised during a surgical aortic valve replacement procedure,which facilitated histopathological examination,biological culture,and high-throughput metagenomic sequencing of pathogenic mi-croorganisms.The definitive diagnosis was established as aortic valve infective endocarditis(Q fever IE)leading by infection of Coxiella burnetii.

Background/Aims: Metabolic dysfunction-associated steatotic liver disease (MASLD) has become an increasingly important health challenge, with a substantial rise linked to changing lifestyles and global obesity. Ursolic acid, a natural pentacyclic triterpenoid, has been explored for its potential therapeutic effects. Given its multifunctional bioactive properties, this research further revealed the pharmacological mechanisms of ursolic acid on MASLD. Methods: Drug target chips and bioinformatics analysis were combined in this study to explore the potential therapeutic effects of ursolic acid on MASLD. Molecular docking simulations, surface plasmon resonance analyses, pull-down experiments, and co-immunoprecipitation assays were used to verify the direct interactions. Gene knockdown mice were generated, and high-fat diets were used to validate drug efficacy. Furthermore, initial CD4+ T cells were isolated and stimulated to demonstrate our findings. Results: In this study, the multifunctional extracellular matrix phosphorylated glycoprotein secreted phosphoprotein 1 (SPP1) was investigated, highlighting its capability to induce Th17 cell differentiation, amplifying inflammatory cascades, and subsequently promoting the evolution of MASLD. In addition, this study revealed that in addition to the canonical TGF-β/IL-6 cytokine pathway, SPP1 can directly interact with ITGB1 and CD44, orchestrating Th17 cell differentiation via their joint downstream ERK signaling pathway. Remarkably, ursolic acid intervention notably suppressed the protein activity of SPP1, suggesting a promising avenue for ameliorating the immunoinflammatory trajectory in MASLD progression. Conclusions Ursolic acid could improve immune inflammation in MASLD by modulating SPP1-mediated Th17 cell differentiation via the ERK signaling pathway, which is orchestrated jointly by ITGB1 and CD44, emerging as a linchpin in this molecular cascade.

To establish an injectable hydrogel containing Prussian blue (PB) nanospheres for photothermal therapy against cancer, PB nanospheres were prepared by one-pot synthesis and the thermosensitive Pluronic F127 was used as the hydrogel matrix. The PB nanospheres and the hydrogel were characterized by shape, particle size, serum stability, photothermal performance upon repeated 808 nm laser irradiation, as well as the rheological features. The effect of the PB nanospheres and the hydrogel were evaluated qualitatively and quantitatively in 4T1 mouse breast cancer cells. The retention, photothermal efficacy, therapeutic effects and systemic toxicity of the hydrogel were assessed in a tumor-bearing mouse model. The PB nanospheres had a diameter of about 150 nm and exhibited satisfactory serum stability, photo-heat convert ability and repeated laser exposure stability. The hydrogel encapsulation did not negatively influence the above features of the photothermal agent. The nanosphere-containing hydrogel showed a phase transition at body temperature and, as a result, a long retention time . The photothermal agent-embedded hydrogel displayed promising photothermal therapeutic effects in the tumor-bearing mouse model with little-to-no systemic toxicity after peritumoral administration.

OBJECTIVE： To establish the method for content determination of total flavonoids from Combretum alfrdii， and to optimize the extraction technology of total flavonoids from C. alfrdii. METHODS： Using aluminium trichloride as， chromogenic agent， UV spectrum was adopted to determine the content of total flavonoids from C. alfrdii. Based on single factor test， ethanol volume fraction， material-liquid ratio， extraction time， extraction temperature and times were selected as investigation factors， and the content of total flavonoids was selected as response value， Plackett-Burman design was used to screen the factors that had significant influence on the content of total flavonoidsfrom C. alfrdii. Then steepest climbing test was adopted to confirm the optimum valuing range； the extraction technology of total flavonoids was optimized by Box-Behnken response methodology. RESULTS： The linear range of total flavonoids were 0.012-0.036 mg/mL （r＝0.999 9）； RSDs of precision， stability and repeatability tests were less than 3％； the recovery ranged from 92.98％ to 99.86％ （RSD＝2.71％， n＝6）. The optimal extraction technology included that 60％ ethanol， material-liquid ratio of 1 ∶ 34 （g/mL）， extracting for 3 times， lasting for 60 min， extraction temperature of 80 ℃. Under this technology， average content of total flavonoids from C. alfrdii was 2.71％ （RSD＝1.69％， n＝6）， and the relative error was 2.65％ compared with predicted value of the model （2.64％）. CONCLUSIONS： Established method is stable and reproducible， and can be used for content determination of total flavonoids from C. alfrdii. The optimized extraction method is stable and feasible.

Objective To investigate the effect ofmiR-184 on proliferation of neural stem cells (NSCs) and its mechanisms in mice.Methods The pHBLV-U6-GFP-miR-184 over-expression plasmid and pHBLV-U6-GFP-miR-184 inhibitor plasmid were used to construct recombinant lentivirus.And the NSCs derived fiom subventricular zone of E14d CD1 mouse were confirmed by immunofluorescence assay.There were four groups that contain a miR-184 overexpression group,a miR-184 inhibitor group and two control groups.The NSCs which infected with lentiviral vectors were selected for puromycin resistance for 5-7 days,and then surviving cells were cultivated to three generations.The expression level ofmiR-184 was detected by real time-quantitative PCR (RT-qPCR).And the target genes ofmiR-184 were predicted through TargetScan,IRTarBase and MiRanda,and were confirmed by Western blotting and RT-qPCR.The cells in the four groups were culttared under proliferating conditions incorporated bromodeoxyuridine (BrdU) in cell proliferation analyses.The protein expressions of Hesl and Hes5,the target proteins of Notch signaling pathways,and their mRNA expressions were detected by Western blotting and RT-qPCR.Results There were 90％ of cells in each group expressing both Nestin and Sox2.The miR-184 level in the miR-184 overexpression group was 67.63±7.53 times of that of the control group,with significant difference (P＜0.05).The percent of BrdU+/DAPI+ of the miR-184 overexpression group was 1.47±0.05 times of that in the control group,with significant difference (P＜0.05);and the percent of BrdU+/DAPI+ of the miR-184 inhibitor group was 0.84±0.03 times of that in the inhibitor control group,with significant difference (P＜0.05).Numbl was a target gene ofmiR-184 indicated by IRTarBase and MiRanda.The miR-184 could inhibit Numbl protein expression;the Numbl protein expression level in the miR-184 overexpression group was 0.73±0.07times of that in the control group,and the Numbl protein expression level in the miR-184 inhibitor group was 1.30±0.05 times of that in the control group,with significant difference (P＜0.05);but miR-184 did not change the Numbl mRNA level.The miR-184 could activate Notch signaling pathway through inhibiting the Numbl protein expression,and increase the Hes1 and Hes5 protein and mRNA expression levels (P＜0.05).Conclusion The miR-184 promotes the NSCs proliferation through inhibiting the Numbl protein translation and further activating the Notch signaling pathway.

OBJECTIVE: To investigate the safety and short-term efficacy of stent on 17 patients with very small intracranial aneurysms.
 METHODS: A total of 17 patients with very small intracranial aneurysms were treated by LVIS stent from October 2014 to November 2015. The location, size of the aneurysms and the branch around aneurysms were evaluated by digital subtraction angiography (DSA). The metal coverage for aneurysms was enhanced by using deployment technology ("compression" mode). The safety and efficacy were assessed after operation.
 RESULTS: LVIS stents-assisted treatments for very small aneurysms were carried out in 17 cases, including 7 cases of paraclinoid aneurysms, 4 cases of posterior communicating artery aneurysms, 3 cases of anterior communicating artery aneurysms, 2 cases of carotid bifurcation aneurysms, 1 case of the superior cerebellar artery aneurysm. The stents for 17 patients with very small intracranial aneurysms were released completely (100%); Raymond grade I embolization was seen in 13 cases (76.5%); Raymond grade II embolization was seen in 4 cases (23.5%); during the follow up from a month to a year, 16 patients showed good curative effect (with the mRS score at 0-2), 1 showed poor effect (with the mRS score at 3-6), and the efficacy rate was 94.1%; no perioperative hemorrhagic and ischemic complications happened.
 CONCLUSION LVIS stent-assisted therapy for very small intracranial aneurysms by using deployment technology was safe and feasible, which can significantly improve the embolization rate for very small aneurysms.
Basilar Artery ; Embolization, Therapeutic ; Humans ; Intracranial Aneurysm ; Safety ; Stents