Objective To explore the roles of the mechanoreceptor Yes-associated protein 1(YAP1)and piezo type mechanosensitive ion channel component 2(PIEZO2)in mechanotransduction in mouse bladder urothelial cells.Methods Mouse bladder urothelial cells were subjected to mechanical stretching using the FX-6000T cell stretching system and treated with the YAP1-specific inhibitor verteporfin(VP).The expressions of PIEZO2,YAP1 and connective tissue growth factor(CTGF)at the mRNA and protein levels,as well as changes in cellular adenosine triphosphatase(ATP)concentration,were detected using reverse transcription quantitative PCR(RT-qPCR)and Western blotting(WB).Results After stretching stimulation,under the fluorescence microscope,it was observed that the diameter length of the stretched cells were longer than that before stretching,and the difference was statistically significant(P＜0.05).The expressions of YAP1,PIEZO2 and CTGF at the mRNA and protein levels were increased in the stretched group compared to those of the non-stretched group(P＜0.05).VP effectively reduced the expressions of YAP1,PIEZO2 and CTGF at the mRNA and protein levels after stretching stimulation(P＜0.05).Stretching stimulation significantly increased the intracellular ATP concentration,while VP was able to inhibit the increase in ATP concentration,with a statistically significant difference(P＜0.000 1).Conclusion Stretching stimulation increased the expressions of YAP1 and PIEZO2 in bladder urothelial cells and promoted the release of ATP;verteporfin inhibited the increase in YAP1 activity and the overexpression of PIEZO2 caused by stretching,thereby reducing the release of ATP.It is suggested that mouse bladder urothelial cells may primarily sense mechanical signals through the YAP1-PIEZO2-ATP pathway.

Objective To explore the risk factors and incidence of epiretinal membrane(ERM)formation following scleral buckling(SB)for rhegmatogenous retinal detachment(RRD),and to construct a risk prediction model to facilitate screening of high-risk populations and prevent ERM formation.Methods RRD patients who underwent SB in the Second Affiliated Hospital of Harbin Medical University between February 2022 and April 2024 were included in the study.The pa-tients were divided into occurrence and non-occurrence groups according to whether they developed ERM.Patient data were analyzed,and univariate Cox regression analysis was performed to select variables,which were then incorporated into the multivariate Cox regression model for the identification of risk factors for ERM formation after SB in RRD patients.A predictive model for ERM risk in RRD patients was constructed based on this data,and nomograms,receiver operating characteristic(ROC)curves,and calibration curves were drawn to evaluate and validate the diagnostic performance of the model.Results A total of 126 RRD patients(126 eyes)who underwent SB were included.There were 27 cases develo-ping ERM(occurrence group)and 99 not developing ERM(non-occurrence group),with an ERM incidence of 21.4％.Multivariate Cox regression analysis revealed that a history of diabetes mellitus[Hazard ratio(HR)=3.52,95％CI:1.37-9.02,P=0.009],preoperative proliferative vitreoretinopathy(PVR)(HR=13.00,95％CI:5.18-32.63,P＜0.001),and ≥4 retinal holes(HR=2.33,95％CI:1.04-5.23,P=0.041)were independent influence factors for ERM formation in RRD patients.ROC curve analysis showed that the area under the curve(AUC)was 0.840(95％CI:0.740-0.940)at 30 days and 0.904(95％CI:0.834-0.975)at 90 days.Conclusion A history of diabetes mellitus,preoperative PVR,and ≥4 retinal holes are factors influencing the development of ERM after SB in RRD patients.It is verified that the risk prediction model constructed based on these factors can accurately predict the risk of ERM formation within 6 months in RRD patients.

Objective:To investigate the role of mechanosensor Yes-associated protein 1 (YAP1) in urothelial cells in inducing bladder dysfunction in a partial bladder outlet obstruction (pBOO) model.Methods:Ten female C57BL/6 mice were included in this study and randomly divided into pBOO and sham groups based on body weight using a stratified pairing method, with 5 mice in each group. The pBOO group underwent proximal urethral ligation surgery, while the sham group underwent a sham operation. Two weeks after surgery, the urinary pattern was analyzed using the urine spot test. The significant increase in urine spot numbers indicated the successful establishment of the pBOO model. The mice were then sacrificed, and bladder tissues were weighed and stained with hematoxylin and eosin (HE) to observe morphological changes. The bladder urothelial layer was further isolated, and total cell proteins were extracted to detect the expression levels of YAP1 protein using Western blotting. Mouse immortalized bladder urothelial cells were divided into three experimental groups: the negative control (NC) group, which was treated with YAP1-NC lentivirus; the overexpression (OE) group, which was treated with YAP1-OE lentivirus to induce YAP1 protein overexpression; and the verteporfin treatment (VP) group, which was treated with verteporfin on the basis of the OE group. Real-time quantitative PCR and Western blotting were used to verify the transcription and expression levels of YAP1 protein, the co-transcriptional activator TEAD4 protein, and the phosphorylated protein DRP1-616 (at serine 616) of dynamin-related protein 1 (DRP1). An ATP detection kit was used to measure the ATP release concentration in the NC, OE, and VP groups. The interaction between YAP1 and TEAD4 was investigated using co-immunoprecipitation, and the expression of the mitochondrial marker translocase of the outer mitochondrial membrane 20 (Tom20) was observed using immunofluorescence staining.Results:The results of the urine spot test showed that the number of urine spots on the filter paper in the pBOO group was higher than that in the sham group within 6 hours [(283.0±9.1) spots vs. (3.7±0.3) spots, P<0.01], and the urine spots were scattered. The bladder wet weight in the pBOO group was significantly higher than that in the sham group [(105.70±6.84) mg vs. (22.33±1.20) mg, P<0.01]. Histological observations revealed reduced bladder mucosal folds and increased detrusor muscle thickness in the pBOO group. The expression of YAP1 protein in the bladder urothelial cells of the pBOO group was significantly upregulated compared to the sham group [(1.26±0.08) vs. (0.50±0.04), P<0.01]. In vitro experiments showed that compared to the NC group, the OE group had significantly increased expression of DRP1-616 [(0.94±0.05) vs. (0.33±0.01), P<0.01] and higher ATP release concentration [(24.45±0.16) μmol/mg vs. (19.67±0.42) μmol/mg, P<0.01]. In contrast, the VP group had significantly decreased expression of DRP1-616 [(0.29±0.04) vs. (0.94±0.05), P<0.01] and lower ATP release concentration [(10.55±0.01) μmol/mg vs. (24.45±0.16) μmol/mg, P<0.01] compared to the OE group. Co-immunoprecipitation experiments using YAP1 and TEAD4 antibodies showed that YAP1 and TEAD4 proteins could interact and form a transcriptional complex to regulate ATP release. Immunofluorescence staining revealed increased expression of Tom20 in the OE group compared to the NC group [(104.20±3.28) vs. (74.51±3.87), P<0.01]. Conclusions:In the pBOO-induced bladder dysfunction model, YAP1 is highly expressed in urothelial cells. YAP1 forms a transcriptional complex with TEAD4 to regulate ATP release by promoting mitochondrial fission via DRP1-616 expression, which is a key mechanism underlying pBOO-induced bladder dysfunction.

Objective To explore the roles of the mechanoreceptor Yes-associated protein 1(YAP1)and piezo type mechanosensitive ion channel component 2(PIEZO2)in mechanotransduction in mouse bladder urothelial cells.Methods Mouse bladder urothelial cells were subjected to mechanical stretching using the FX-6000T cell stretching system and treated with the YAP1-specific inhibitor verteporfin(VP).The expressions of PIEZO2,YAP1 and connective tissue growth factor(CTGF)at the mRNA and protein levels,as well as changes in cellular adenosine triphosphatase(ATP)concentration,were detected using reverse transcription quantitative PCR(RT-qPCR)and Western blotting(WB).Results After stretching stimulation,under the fluorescence microscope,it was observed that the diameter length of the stretched cells were longer than that before stretching,and the difference was statistically significant(P＜0.05).The expressions of YAP1,PIEZO2 and CTGF at the mRNA and protein levels were increased in the stretched group compared to those of the non-stretched group(P＜0.05).VP effectively reduced the expressions of YAP1,PIEZO2 and CTGF at the mRNA and protein levels after stretching stimulation(P＜0.05).Stretching stimulation significantly increased the intracellular ATP concentration,while VP was able to inhibit the increase in ATP concentration,with a statistically significant difference(P＜0.000 1).Conclusion Stretching stimulation increased the expressions of YAP1 and PIEZO2 in bladder urothelial cells and promoted the release of ATP;verteporfin inhibited the increase in YAP1 activity and the overexpression of PIEZO2 caused by stretching,thereby reducing the release of ATP.It is suggested that mouse bladder urothelial cells may primarily sense mechanical signals through the YAP1-PIEZO2-ATP pathway.

Objective To explore the risk factors and incidence of epiretinal membrane(ERM)formation following scleral buckling(SB)for rhegmatogenous retinal detachment(RRD),and to construct a risk prediction model to facilitate screening of high-risk populations and prevent ERM formation.Methods RRD patients who underwent SB in the Second Affiliated Hospital of Harbin Medical University between February 2022 and April 2024 were included in the study.The pa-tients were divided into occurrence and non-occurrence groups according to whether they developed ERM.Patient data were analyzed,and univariate Cox regression analysis was performed to select variables,which were then incorporated into the multivariate Cox regression model for the identification of risk factors for ERM formation after SB in RRD patients.A predictive model for ERM risk in RRD patients was constructed based on this data,and nomograms,receiver operating characteristic(ROC)curves,and calibration curves were drawn to evaluate and validate the diagnostic performance of the model.Results A total of 126 RRD patients(126 eyes)who underwent SB were included.There were 27 cases develo-ping ERM(occurrence group)and 99 not developing ERM(non-occurrence group),with an ERM incidence of 21.4％.Multivariate Cox regression analysis revealed that a history of diabetes mellitus[Hazard ratio(HR)=3.52,95％CI:1.37-9.02,P=0.009],preoperative proliferative vitreoretinopathy(PVR)(HR=13.00,95％CI:5.18-32.63,P＜0.001),and ≥4 retinal holes(HR=2.33,95％CI:1.04-5.23,P=0.041)were independent influence factors for ERM formation in RRD patients.ROC curve analysis showed that the area under the curve(AUC)was 0.840(95％CI:0.740-0.940)at 30 days and 0.904(95％CI:0.834-0.975)at 90 days.Conclusion A history of diabetes mellitus,preoperative PVR,and ≥4 retinal holes are factors influencing the development of ERM after SB in RRD patients.It is verified that the risk prediction model constructed based on these factors can accurately predict the risk of ERM formation within 6 months in RRD patients.

Objective:To investigate the role of mechanosensor Yes-associated protein 1 (YAP1) in urothelial cells in inducing bladder dysfunction in a partial bladder outlet obstruction (pBOO) model.Methods:Ten female C57BL/6 mice were included in this study and randomly divided into pBOO and sham groups based on body weight using a stratified pairing method, with 5 mice in each group. The pBOO group underwent proximal urethral ligation surgery, while the sham group underwent a sham operation. Two weeks after surgery, the urinary pattern was analyzed using the urine spot test. The significant increase in urine spot numbers indicated the successful establishment of the pBOO model. The mice were then sacrificed, and bladder tissues were weighed and stained with hematoxylin and eosin (HE) to observe morphological changes. The bladder urothelial layer was further isolated, and total cell proteins were extracted to detect the expression levels of YAP1 protein using Western blotting. Mouse immortalized bladder urothelial cells were divided into three experimental groups: the negative control (NC) group, which was treated with YAP1-NC lentivirus; the overexpression (OE) group, which was treated with YAP1-OE lentivirus to induce YAP1 protein overexpression; and the verteporfin treatment (VP) group, which was treated with verteporfin on the basis of the OE group. Real-time quantitative PCR and Western blotting were used to verify the transcription and expression levels of YAP1 protein, the co-transcriptional activator TEAD4 protein, and the phosphorylated protein DRP1-616 (at serine 616) of dynamin-related protein 1 (DRP1). An ATP detection kit was used to measure the ATP release concentration in the NC, OE, and VP groups. The interaction between YAP1 and TEAD4 was investigated using co-immunoprecipitation, and the expression of the mitochondrial marker translocase of the outer mitochondrial membrane 20 (Tom20) was observed using immunofluorescence staining.Results:The results of the urine spot test showed that the number of urine spots on the filter paper in the pBOO group was higher than that in the sham group within 6 hours [(283.0±9.1) spots vs. (3.7±0.3) spots, P<0.01], and the urine spots were scattered. The bladder wet weight in the pBOO group was significantly higher than that in the sham group [(105.70±6.84) mg vs. (22.33±1.20) mg, P<0.01]. Histological observations revealed reduced bladder mucosal folds and increased detrusor muscle thickness in the pBOO group. The expression of YAP1 protein in the bladder urothelial cells of the pBOO group was significantly upregulated compared to the sham group [(1.26±0.08) vs. (0.50±0.04), P<0.01]. In vitro experiments showed that compared to the NC group, the OE group had significantly increased expression of DRP1-616 [(0.94±0.05) vs. (0.33±0.01), P<0.01] and higher ATP release concentration [(24.45±0.16) μmol/mg vs. (19.67±0.42) μmol/mg, P<0.01]. In contrast, the VP group had significantly decreased expression of DRP1-616 [(0.29±0.04) vs. (0.94±0.05), P<0.01] and lower ATP release concentration [(10.55±0.01) μmol/mg vs. (24.45±0.16) μmol/mg, P<0.01] compared to the OE group. Co-immunoprecipitation experiments using YAP1 and TEAD4 antibodies showed that YAP1 and TEAD4 proteins could interact and form a transcriptional complex to regulate ATP release. Immunofluorescence staining revealed increased expression of Tom20 in the OE group compared to the NC group [(104.20±3.28) vs. (74.51±3.87), P<0.01]. Conclusions:In the pBOO-induced bladder dysfunction model, YAP1 is highly expressed in urothelial cells. YAP1 forms a transcriptional complex with TEAD4 to regulate ATP release by promoting mitochondrial fission via DRP1-616 expression, which is a key mechanism underlying pBOO-induced bladder dysfunction.

Objective:To investigate the clinicopathological features, immunophenotype, diagnosis and differential diagnosis of SRF-rearranged cellular perivascular myoid tumor.Methods:Two cases of SRF-rearranged cellular perivascular myoid tumor diagnosed in the Department of Pathology, Fudan University Shanghai Cancer Center from October 2021 to March 2022 were collected. Immunohistochemical staining, fluorescence in-situ hybridization (FISH) and next-generation sequencing (NGS) were performed, and the literature was reviewed.Results:Case 1, a 3-month-old boy presented with a painless tumor of the scalp, measuring about 2 cm in diameter. Case 2, a 3-year-old girl complained with a painless tumor of the knee, measuring approximately 1.5 cm in diameter. Microscopically, the tumor had a clear boundary and showed multinodular growth. The tumor was mainly composed of spindle cells arranged in long intersecting fascicles associated with thin, slit-like or branching ectatic vessels, focally forming hemangiopericytoma-like appearance. The tumor cells were abundant, but there was no obvious atypia. Mitotic figures (3-4/10 HPF) were noted. H-caldesmon and SMA were positive in both cases. Case 1 showed diffuse and strong positivity for Desmin, and focally for CKpan. Ki-67 proliferation index was 20% and 30%, respectively. FISH displayed NCOA2 gene translocation in case 1 and the RELA gene translocation in case 2. NGS detected the SRF-NCOA2 gene fusion in case 1 and the SRF-RELA gene fusion in case 2. Both patients underwent local excisions. During the follow-up of 5-14 months, case 1 had no local recurrence, while case 2 developed local recurrence 1 year post operatively.Conclusions:SRF-rearranged cellular perivascular myoid tumor is a novel variant of perivascular cell tumor, which tends to occur in children and adolescents. The tumor forms a broad morphologic spectrum ranging from a pericytic pattern to a myoid pattern, and include hybrid tumors with a mixture of pericytic and myoid patterns. Due to its diffuse hypercellularity and increased mitotic figures and smooth muscle-like immunophenotype, the tumor is easy to be misdiagnosed as myogenic sarcomas. The tumor usually pursues a benign clinical course and rare cases may locally recur.

Objective:To investigate the clinicopathological features, pathologic diagnosis and differential diagnosis of florid vascular proliferation (FVP) of the intestinal tract.Methods:Ten cases of FVP of the intestinal tract diagnosed from 2010 to 2020 at Fudan University Shanghai Cancer Center were collected. The histomorphology and immunohistochemical staining were evaluated and the relevant literature was reviewed.Results:There were five males and five females, aging from 28 to 76 years (mean 51.0 years; median 50.5 years). Five cases occurred in the colon, three cases in the small intestine, and one each case in the inguinal region and cecum. Clinically, the patients mainly presented with abdominal pain, diarrhea and hematochezia. Seven of nine patients with imaging data showed associated intussusception. Microscopically, the lesion presented lobular growth of florid proliferation of small vessels extending through the bowel wall. The vascular channels were lined with bland endothelial cells with no nuclear atypia and infrequent mitoses. The overlying mucosa showed chronic ulceration. Immunohistochemically, endothelial cells of FVP were positive for CD31, CD34, ERG and Fli1, the stromal spindle cells expressed SMA, and the Ki-67 proliferation index was low (5%-30%). None of 4 patients with follow-up information had local recurrence.Conclusions:FVP is a rare benign vascular proliferation lesion which often occurs in the intestinal tract and is associated with intussusception. Accurate pathologic diagnosis of FVP requires close combination of radiological examinations. FVP is easily misdiagnosed as a true vascular tumor, especially angiosarcoma. It is necessary to better understand FVP to avoid misdiagnosis.

Objective:To analyze the changes of helper T cell 22 (Th22) and related cytokines and chemokines in patients with IgA nephropathy (IgAN) and tonsillitis, and explore its relationship with clinical pathological changes.Methods:IgAN patients who were diagnosed at the Xiangya Hospital of Central South University from June 2015 to June 2016 were included. They were divided into IgAN and tonsillitis (IgAN+tonsillitis) group, IgAN group, mesangial proliferative glomerulonephritis (MsPGN) group and control group (HC) group according to renal pathology and whether associated with tonsillitis. Flow cytometry was used to detect the percentage of peripheral blood Th17, Th22 cells, CC-type chemokine receptor (CCR) 4, CCR6 and CCR10 cells. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of interleukin (IL)-22, IL-1β, IL-6, TNF-α, CCL22, CCL20 and CCL27. Immunohistochemical method (IHC) was used to detect the expression of CCL22, CCL20 and CCL27 in kidney tissue. The differences of clinicopathological indicators, the proportion of Th22 cells and related chemokine in each group were compared and analyzed.Results:A total of 44 IgAN patients were included, including 14 patients complicated with tonsillitis. Ten MsPGN patients and 16 healthy people were also included. There was no statistically significant difference in gender, age, blood pressure, kidney function, blood lipid and other biochemical indicators among the groups (all P>0.05). The peripheral blood Th22 cells and CCR10 positive cells in the IgAN group, MsPGN group, and IgAN+tonsillitis group were significantly higher than those of the control group, and serum IL-22, IL-1β, IL-6, TNF-α, CCL20, CCL22 and CCL27 levels were also significantly higher (all P<0.05). All above indexes reached the highest levels in IgAN patients combined with tonsillitis. The changes of CCL20, CCL22 and CCL27 in renal tissues were consistent with those in peripheral blood. The percentage of Th22 cells increased in hematuria-positive and higher MEST scores patients. Conclusions:Th22 cells cooperated with CCL27/CCR10 axis are involved in the pathogenesis of IgAN. Tonsillitis exacerbates clinical severity and kidney injury of IgAN.