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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 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.

Chronic myeloid leukemia (CML) is a slowly progressing hematopoietic cell disorder. Sphingosine kinase 1 (SPHK1) plays established roles in tumor initiation, progression, and chemotherapy resistance in a wide range of cancers, including leukemia.However, small-molecule inhibitors targeting SPHK1 in CML still need to be developed. This study revealed the role of SPHK1 in CML and investigated the potential anti-leukemic activity of hirsuteine (HST), an indole alkaloid obtained from the oriental plant Uncaria rhynchophylla, in CML cells. These results suggest that SPHK1 is highly expressed in CML cells and that overexpression of SPHK1 represents poor clinical outcomes in CML patients. HST exposure led to G2/M phase arrest, cellular apoptosis, and downregulation of Cyclin B1 and CDC2 and cleavage of Caspase 3 and PARP in CML cells. HST shifted sphingolipid rheostat from sphingosine 1-phosphate (S1P) towards the ceramide coupled with a marked inhibition of SPHK1. Mechanistically, HST significantly blocked SPHK1/S1P/S1PR1 and BCR-ABL/PI3K/Akt pathways. In addition, HST can be docked with residues of SPHK1 and shifts the SPHK1 melting curve, indicating the potential protein-ligand interactions between SPHK1 and HST in both CML cells. SPHK1 overexpression impaired apoptosis and proliferation of CML cells induced by HST alone. These results suggest that HST, which may serve as a novel and specific SPHK1 inhibitor, exerts anti-leukemic activity by inhibiting the SPHK1/S1P/ S1PR1 and BCR-ABL/PI3K/Akt pathways in CML cells, thus conferring HST as a promising anti-leukemic drug for CML therapy in the future.

Cutaneous T-cell lymphomas are a relatively rare group of mature T-cell lymphomas mainly manifesting in the skin, and its common subtype is mycosis fungoides. Total skin electron irradiation is one of the important conventional treatment methods, but there are many disadvantages, such as uneven dose distribution, poor position repetition, and long treatment time, which affect the clinical efficacy and patient prognosis. With the emergence and gradual popularization of helical tomotherapy in recent years, more and more medical institutions are gradually expanding their applications in total skin irradiation due to their ability to treat ultra-long targets and achieve dose-sculpted distribution, aiming to further explore its good or bad, and confirm whether it can replace the traditional total skin electron irradiation. In this article, research progress on total skin irradiation using helical tomotherapy was reviewed, the development of treatment technology, clinical efficacy and current concerns and controversies were illustrated.

Objective:To study the changing characteristics and impact factors of helical tomotherapy (HT)for longitudinal dose fall-off outside the target, in order to guide the plan junction or pretreatment target and implementation efficiency in clinical.Methods:Eight patients with head and neck tumors admitted to the Department of Oncology Radiotherapy of the First Affiliated Hospital of Zhengzhou University in December 2019 were retrospectively selected as the research objects. The planning target area and dose drop structure were outlined in the head and neck images with a thickness of 1 mm obtained by Siemens SOMATOM Definition AS positioning computerized tomography (CT). Different field widths (FW, 5.0 cm/2.5 cm/1.0 cm) and pitches (0.430/0.287/0.215) were assembled for planning with the same modulation factor (1.8), finest does calculation grid (0.195 cm ×0.195 cm) and other planning parameters were consistent. The plans were designed by different parameters, and the result was analyzed by univariate analysis.Results:The that different pitch curves coincided under the same field width by comparative analyzing, so pitchs had no effect on dose drop. The different field width curves were independent of each other, indicating that the field width had an effect on dose drop in the head and foot direction. The relationship between the longitudinal dose drop speed outside the target and the change of the field width was inversely correlated: the larger field widths meant the slower dose fall-off and the larger penumbra, while the smaller field widths meant the faster fall-off and the smaller penumbra. When the dose fall-off to 50% of the prescribed dose, the distance from the target was approximately equal to half the field widths, and the pitchs had not affect the rate of dose-drop, while the dose at different distances from the target boundary could be calculated by the fitting formulas. The field widths and pitchs had little effect on the CI and HI index of the target, relatively, the target area was best when the field width was 2.5 cm. The total beam-on time gradually decreased with the increase of the field widths and pitches.Conclusions:When segment target therapy needs to consider planning junction, execution efficiency, and controlling longitudinal dose fall-off and considered the execution, the optimal planned parameters such as field widths and pitches could be selected or the target at the junction regions could be adducted according to the longitudinal dose drop formula, so as to achieve the ideal dose distribution.

Objective:To investigate the effect of different ways of reconstruction of left subclavian artery (LSA) in the treatment of complex aortic arch lesions.Methods:The clinical data of 34 patients with complex thoracic aortic disease undergoing intracavitary LSA reconstruction in our center from January 2019 to February 2020 were retrospectively analyzed. The distance of proximal healthy landing zone of all patients, including 29 aortic dissections involving LSA, 3 penetrating aortic ulcer and 2 thoracic aortic aneurysms, was less than 15 mm. Among them, 16 cases were treated with chimney technique, 16 cases were implanted with single branched stent-graft, 2 cases were received with left common carotid artery and LSA in situ fenestration.Results:The operation success rate of all 34 patients was 100%. One case was changed from in situ fenestration to chimney stenting. Followed up for 1-12 months, there were no death, cerebral ischemia, paraplegia and other postoperative complications. CTA review showed that the main and branch stents were in good shape, the patency rate of LSA branch stents was 100% and no endoleak occurred at 1 and 3 months after operation. The muscle strength and arterial blood pressure of bilateral upper limbs of all patients were basically the same.Conclusion:There is no consensus for the treatment of complex aortic arch lesions, so we need to customize the personalized plan and select the appropriate LSA reconstruction method in order to reduce the incidence of complications.

Objective To summarize the clinical efficacy of thoracic endovascular aortic repair (TEVAR) in the treatment of Stanford type B thoracic aortic dissection. Methods The clinical data of 80 patients with Stanford type B aortic dissection who had underwent TEVAR in cardiac surgery of the First Affiliated Hospital of University of Science and Technology of China from January 2017 to December 2017 were analyzed retrospectively. Among them, there were 56 males and 24 females. The effect of operation and postoperative complications were observed. The diameters of different aortic levels before and after TEVAR were compared in order to understand the aortic remodeling after TEVAR. Results All 80 patients were operated successfully. A total of 87 stents were implanted, of which 2 stents were placed in 7 patients. Four patients died 30 days after operation, 3 of whom were diagnosed as dissection rupture before operation and underwent emergency TEVAR. The cause of death was massive hemorrhage due to re-rupture of dissection. One patient complicated with massive cerebral infarction before operation died of respiratory failure. Six months after operation, CTA showed that the diameter of the aortic true lumen at the level of the left subclavian artery, the maximum diameter of the tumor and the level of the diaphragm significantly increased: (30.1 ± 3.5) mm vs. (24.4 ± 4.2)mm, (33.4 ± 5.1) mm vs. (24.9 ± 6.2) mm,(26.1±4.9) mm vs. (19.3 ± 3.1) mm; all P values＜0.01, and the false lumen significantly decreased: (3.5 ± 1.7) mm vs. (11.2 ± 5.7) mm, (9.1 ± 2.4) mm vs. (18.3 ± 5.9) mm, (6.2 ± 1.3) mm vs. (14.7 ± 5.2) mm, all P values＜0.01. There was no significant difference in the overall diameter of aorta before and after operation (P > 0.05). Conclusions The treatment of Stanford type B aortic dissection with TEVAR has significantly good short-term clinical efficacy, which is less traumatic and quick to recover. However, the long-term efficacy remains to be observed.

Objective To externally validate the accuracy of combined use of neutrophil-lymphocyte ratio (NLR),V20,and Dmean in predicting the incidence of grade Ⅲ or higher radiation-induced lung injury (RILI) in lung cancer patients.Methods A total of 166 lung cancer patients,who participated in the model establishment were selected into the internal validation group,and 85 lung cancer patients who received intensity-modulated radiotherapy in our department between June 2016 and June 2018 were assigned into the external validation group.The incidence rate of grade 3 or higher RILI was statistically compared between the internal and external validation groups.Multivariate logistic analysis was performed for NLR,V20 and Dmean The discrimination degree of the predictive model was evaluated by using ROC curve in combination with NLR,V20 and Dmean The calibration degree of the predictive model was assessed by Hosmer-Lemeshow test.Results The incidence rate of grade 3 or higher RILI in the internal and external validation groups was 23.8％ and 22.9％.Multivariate logistic analysis demonstrated that NLR,V20 and Dmean significantly differed in the internal validation group (P=0.032,0.006 and 0.005).However,only V20 significantly differed in the external validation group (P=0.038).The discrimination and calibration degree of RILI was almost consistent between the internal and external validation groups (both P＞0.05).The area under the curve (AUC) predicted by NLR,V20,Dmean and the combination of three indexes were 0.611,0.646,0.682 and 0.775 in the internal validation group,and 0.544,0.702,0.658 and 0.754 in the external validation group,respectively.The calibration degree in the internal validation group was P=2.797and 0.834,P=2.452 and 0.653 in the external validation group.Conclusion Combined application of NLR,V20 and Dmean can accurately predict the incidence of grade Ⅲ or higher RILI in lung can cancer patients,which has been validated by external dataset.

Objective To evaluate the feasibility of total marrow and lymphatic irradiation (TMLI)with helical tomotherapy as a conditioning regimen before hematopoietic stem cell transplantation (HSCT).Methods Seven children with acute lymphoblastic leukemia and aplastic anemia were recruited as study subjects.The median age was 7 years old.The prescribed dose was 12 Gy/6 fractions twice daily.The exposure dose of the target and the organs at risk between helical helical tomotherapy-based TMLI regimen and total body irradiation (TBI) regimen were statistically compared,and acute toxicity grading was performed for all patients.Results Compared with the TBI regimen,the average exposure dose reduction for organs at risk after the TMLI regimen was ranged from 4.2％ to 40.6％.The average exposure dose reduction for the kidney was the largest among all organs.The acute toxicities experienced by all patients were graded and recorded including 2 cases of nausea,5 cases of vomiting,1 case of anorexia,1 case of eryhema,3 cases of diarrhea,and 1 case of oral mucositis.Only grade 1-2 toxicities were observed,and no grade 3-4 toxicities occurred.Conclusions The findings in this study confirm the feasibility of helical helical tomotherapy-based TMLI regimen.Compared with the TBI regimen,the mean duration of treatment for the TMLI regimen with an equivalent dose is not increased.The exposure dose experienced by organs at risk is reduced and the predicted incidence rate is decreased when the TMLI regimen is employed,which provides a myeloablative pretreatment strategy.However,the long-term toxicity of TMLI regime remains to be evaluated by clinical trials.