Corticotomy is a clinical procedure to accelerate orthodontic tooth movement characterized by the regional acceleratory phenomenon (RAP). Despite its therapeutic effects, the surgical risk and unclear mechanism hamper the clinical application. Numerous evidences support macrophages as the key immune cells during bone remodeling. Our study discovered that the monocyte-derived macrophages primarily exhibited a pro-inflammatory phenotype that dominated bone remodeling in corticotomy by CX3CR1^CreERT2; R26^GFP lineage tracing system. Fluorescence staining, flow cytometry analysis, and western blot determined the significantly enhanced expression of binding immunoglobulin protein (BiP) and emphasized the activation of sensor activating transcription factor 6 (ATF6) in macrophages. Then, we verified that macrophage specific ATF6 deletion (ATF6^f/f; CX3CR1^CreERT2 mice) decreased the proportion of pro-inflammatory macrophages and therefore blocked the acceleration effect of corticotomy. In contrast, macrophage ATF6 overexpression exaggerated the acceleration of orthodontic tooth movement. In vitro experiments also proved that higher proportion of pro-inflammatory macrophages was positively correlated with higher expression of ATF6. At the mechanism level, RNA-seq and CUT&Tag analysis demonstrated that ATF6 modulated the macrophage-orchestrated inflammation through interacting with Tnfα promotor and augmenting its transcription. Additionally, molecular docking simulation and dual-luciferase reporter system indicated the possible binding sites outside of the traditional endoplasmic reticulum-stress response element (ERSE). Taken together, ATF6 may aggravate orthodontic bone remodeling by promoting Tnfα transcription in macrophages, suggesting that ATF6 may represent a promising therapeutic target for non-invasive accelerated orthodontics.
Animals ; Mice ; Macrophages/metabolism* ; Tumor Necrosis Factor-alpha/genetics* ; Tooth Movement Techniques/methods* ; Activating Transcription Factor 6/metabolism* ; Bone Remodeling ; Flow Cytometry ; Blotting, Western

Objective:This study aimed to evaluate the prognostic factors of clinical and histopathological parameters,including age,gender,tumor stage,tumor grade,tumor differentiation,lymph node metastasis(LNM),tumor frequency,and tumor count,in patients undergoing radical cystectomy(RC)combined with Mainz Pouch Ⅱ bladder reconstruction.Methods:A total of 237 bladder cancer patients(198 male and 39 female)who underwent RC combined with Mainz Pouch Ⅱ bladder reconstruction without chemotherapy or radiotherapy,from January 2004 to January 2023,were included in this study.Kaplan-Meier and Cox regression analyses were performed to assess the impact of age,tumor grade,tumor stage,tumor differentiation,LNM,tumor frequency,and tumor count on 5-year overall survival(OS)and 5-year cancer-specific survival(CSS).Results:The mean age at diagnosis was 59.8 years,with 198 male and 39 female patients.The mean follow-up duration was 47.8 months.In univariate analysis,patients younger than 65 years had significantly higher 5-year OS and 5-year CSS compared to those aged≥65 years.Patients with urothelial carcinoma showed better 5-year OS than those with non-urothelial carcinoma.Additionally,tumor stage,tumor grade,and LNM were negatively associated with 5-year OS and 5-year CSS.On multivariate analysis,only tumor grade and LNM remained statistically significant(P<0.05).Conclusions:Tumor grade and LNM were identified as independent prognostic risk factors for 5-year OS and 5-year CSS following RC combined with Mainz PouchⅡ bladder reconstruction.Moreover,the application of RC combined with Mainz Pouch Ⅱ bladder reconstruction should consider the patient's preferences and physical condition.

Objective To identify genetic regulators of ionizing radiation(IR)sensitivity through clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated nuclease 9(Cas9)genome-wide screening.Methods A specialized single guide RNA(sgRNA)library was developed targeting 987 stress-response regulatory genes identified through Kyoto Encyclopedia of Genes and Genomes(KEGG),Reactome and gene ontology(GO)databases,followed by construction of sgRNA plasmids and packaging into a lentiviral library.Using colon adenocarcinoma Caco-2 cells as a model system,the Cas9-stable transgenic line was established via lentiviral transduction and antibiotic selection.Library-transduced cells underwent puromycin selection,followed by γ-irradiation(dose optimized via preliminary experiments).Post-irradiation phenotypic selection was conducted after 14 days,with subsequent next-generation sequencing of surviving cell populations to identify enriched/depleted sgRNAs.Candidate genes were functionally validated through orthogonal assays:cell counting kit-8(CCK-8)proliferation analysis,clonogenic survival assays and flow cytometric quantification of reactive oxygen species(ROS)and apoptotic markers.Results The optimized screening platform identified 281 radiation response genes(165 radiosensitive and 116 radioprotective candidates).Functional validation revealed Bcl2-associated athanogene 3(BAG3)knockdown significantly enhanced radioresistance.Proliferation was increased 1.2-1.5 fold,clonogenic survival improved 1.5-2.0 fold,and ROS was reduced by 13％-25％coupled with 32％-73％apoptosis attenuation compared to control groups.Conclusion The integrated CRISPR/Cas9 screening platform effectively identifies novel radiation response regulators,as evidenced by the discovery of BAG3 as a critical radiosensitivity determinant.This high-throughput functional genomics approach provides a robust methodology for systematically mapping molecular determinants of cellular radiation response,with potential applications in both mechanistic studies and therapeutic target discovery.

Objective To analyze the clinical value of plasma insulin-like factor 6(INSL6)in predicting the risk of acute aortic syndromes(AAS)and adverse outcomes.Methods A retrospective case-control trial was conducted on 194 AAS patients admitted to Department of Cardiovascular Diseases of Second Affiliated Hospital of Army Medical University between April 2021 and June 2023.Another 194 sex-,age-and BMI-matched individuals without aortic diseases were recruited from the health examination center between December 2021 and January 2024.Their plasma INSL6 level was measured with ELISA,and the general clinical data and results of some laboratory tests were collected and compared between the 2 groups.Spearman correlation analysis was used to assess the relationship between plasma INSL6 and other variables,receiver operating characteristic(ROC)curve was plotted to evaluate the diagnostic efficacy of INSL6 for AAS occurrence,multivariate conditional logistic regression model was utilized to analyze the association between plasma INSL6 and AAS onset,and Kaplan-Meier survival curve and multivariate Cox proportional hazards regression model was applied to analyze the relationship between acute-phase plasma INSL6 level and adverse prognosis in AAS patients.Results The plasma INSL6 level was significantly higher in AAS patients at acute phase than the control individuals[704.40(481.32～1 152.62)vs 141.24(107.60～163.72)pg/mL,P<0.001],but no statistical difference was observed in the level among the patients with different AAS subtypes(aortic dissection,intramural hematoma,and penetrating aortic ulcer).Spearman correlation analysis showed that the plasma INSL6 level was positively correlated with platelet count(r=0.325,P<0.001)and hemoglobin concentration(r=0.186,P=0.009),and negatively with IL-6(r=-0.182,P=0.011),INF-γ(r=-0.283,P<0.001),and D-dimer levels(r=-0.195,P=0.006).Multivariate conditional logistic regression analysis revealed that plasma INSL6 level was independently associated with the occurrence of AAS(OR=28.634,95%CI:7.267～112.820,P<0.001).ROC curve analysis further confirmed that the optimal cutoff value of plasma INSL6 in predicting AAS was 259.425 pg/mL,with a sensitivity of 95.9%and a specificity of 98.5%at this threshold.Kaplan-Meier curve analysis demonstrated that AAS patients with low INSL6 level had significantly lower cumulative survival rates(P=0.020)and event-free survival rates(P=0.004)than those with high INSL6 level(P<0.05).Multivariate COX regression analysis revealed that,after adjusting for sex,age,systolic blood pressure,ST classification,and surgical treatment,acute-phase INSL6 level was independently associated with all-cause mortality(HR:0.999,95%CI:0.999～1.000,P=0.023),AAS-related mortality(HR:0.999,95%CI:0.998～1.000,P=0.012),and composite endpoint events(HR:0.999,95%CI:0.999～1.000,P=0.026)in AAS patients during follow-up.Conclusion Plasma INSL6 level is closely associated with the occurrence and adverse prognosis of AAS,and the indicator is expected to be an effective biomarker for diagnosing AAS and predicting its prognosis.

Objective:To evaluate the feasibility and safety of a fascial space priority approach to total pelvic exenteration (TPE) in patients with pelvic malignancy.Methods:This was a descriptive case series. Relevant clinical data of patients who had undergone TPE via a fascial space priority approach at Tianjin Union Medical Center from September 2017 to March 2025 were retrospectively collected. All operations had been performed via a fascial space priority approach, the guiding principle of which is separating the avascular pelvic spaces first and then transecting the vessels and nerves of the pelvic organs. That is, the avascular planes around all the pelvic organs are dissected first, after which the relevant vessels and nerves are fully dissected and transected, followed by en bloc resection of pelvic organs distally or via perineal approach. The variables studied included relevant surgical parameters, postoperative pathological findings, complications (classified according to the Clavien-Dindo criteria); recurrence-free survival (RFS), overall survival, and tumor-specific survival. Results:The study cohort comprised 41 patients, including 30 (73.2%) with primary tumors and 11 (26.8%) with recurrent tumors. Open TPE was performed on five patients (12.2%) and laparoscopic TPE on the remaining 36 (87.8%). All procedures were successfully completed with a fascial space priority approach and there were no intraoperative deaths. R0 resection was achieved in 34 patients (82.9%) and R1 resection in seven (17.1%). The operation time was 500 (265-740) min, and the amount of bleeding 200 (10-3,500) mL. Twelve patients (29.3%) developed postoperative complications, two of which were Clavien-Dindo Grade III complications. One of these patients required re-operation to manage a pelvic hematoma 29 days after the primary TPE. No active bleeding was observed during the re-operation. Another patient underwent interventional angiography for an episode of postoperative bleeding; this showed a pseudoaneurysm of the internal iliac artery that was successfully treated by interventional embolization via the internal iliac artery. Five days after undergoing a primary TPE with bladder preservation, a third patient was found to have a urinary fistula and underwent laparoscopic bladder resection with percutaneous ureterostomy. The median duration of follow-up was 18 (1-90) months. The 5-year RFS and overall survival were 46.7% and 52.2%, respectively, whereas the 5-year tumor-specific survival was 67.8%. Univariate Cox regression analysis identified a positive surgical margin ( P < 0.001), lateral pelvic sidewall invasion ( P=0.014), and vascular invasion ( P=0.004) as significantly associated with RFS, whereas multivariate analysis identified only a positive surgical margin (HR: 21.93, 95% CI: 3.78-127.42, P<0.001) as an independent predictor of RFS. Conclusions:It is safe and feasible to perform TPE with a fascial space priority approach on patients with pelvic malignancy. Positive surgical margins are significantly associated with RFS.

Objective:To evaluate the feasibility and safety of a fascial space priority approach to total pelvic exenteration (TPE) in patients with pelvic malignancy.Methods:This was a descriptive case series. Relevant clinical data of patients who had undergone TPE via a fascial space priority approach at Tianjin Union Medical Center from September 2017 to March 2025 were retrospectively collected. All operations had been performed via a fascial space priority approach, the guiding principle of which is separating the avascular pelvic spaces first and then transecting the vessels and nerves of the pelvic organs. That is, the avascular planes around all the pelvic organs are dissected first, after which the relevant vessels and nerves are fully dissected and transected, followed by en bloc resection of pelvic organs distally or via perineal approach. The variables studied included relevant surgical parameters, postoperative pathological findings, complications (classified according to the Clavien-Dindo criteria); recurrence-free survival (RFS), overall survival, and tumor-specific survival. Results:The study cohort comprised 41 patients, including 30 (73.2%) with primary tumors and 11 (26.8%) with recurrent tumors. Open TPE was performed on five patients (12.2%) and laparoscopic TPE on the remaining 36 (87.8%). All procedures were successfully completed with a fascial space priority approach and there were no intraoperative deaths. R0 resection was achieved in 34 patients (82.9%) and R1 resection in seven (17.1%). The operation time was 500 (265-740) min, and the amount of bleeding 200 (10-3,500) mL. Twelve patients (29.3%) developed postoperative complications, two of which were Clavien-Dindo Grade III complications. One of these patients required re-operation to manage a pelvic hematoma 29 days after the primary TPE. No active bleeding was observed during the re-operation. Another patient underwent interventional angiography for an episode of postoperative bleeding; this showed a pseudoaneurysm of the internal iliac artery that was successfully treated by interventional embolization via the internal iliac artery. Five days after undergoing a primary TPE with bladder preservation, a third patient was found to have a urinary fistula and underwent laparoscopic bladder resection with percutaneous ureterostomy. The median duration of follow-up was 18 (1-90) months. The 5-year RFS and overall survival were 46.7% and 52.2%, respectively, whereas the 5-year tumor-specific survival was 67.8%. Univariate Cox regression analysis identified a positive surgical margin ( P < 0.001), lateral pelvic sidewall invasion ( P=0.014), and vascular invasion ( P=0.004) as significantly associated with RFS, whereas multivariate analysis identified only a positive surgical margin (HR: 21.93, 95% CI: 3.78-127.42, P<0.001) as an independent predictor of RFS. Conclusions:It is safe and feasible to perform TPE with a fascial space priority approach on patients with pelvic malignancy. Positive surgical margins are significantly associated with RFS.

Objective:This study aimed to evaluate the prognostic factors of clinical and histopathological parameters,including age,gender,tumor stage,tumor grade,tumor differentiation,lymph node metastasis(LNM),tumor frequency,and tumor count,in patients undergoing radical cystectomy(RC)combined with Mainz Pouch Ⅱ bladder reconstruction.Methods:A total of 237 bladder cancer patients(198 male and 39 female)who underwent RC combined with Mainz Pouch Ⅱ bladder reconstruction without chemotherapy or radiotherapy,from January 2004 to January 2023,were included in this study.Kaplan-Meier and Cox regression analyses were performed to assess the impact of age,tumor grade,tumor stage,tumor differentiation,LNM,tumor frequency,and tumor count on 5-year overall survival(OS)and 5-year cancer-specific survival(CSS).Results:The mean age at diagnosis was 59.8 years,with 198 male and 39 female patients.The mean follow-up duration was 47.8 months.In univariate analysis,patients younger than 65 years had significantly higher 5-year OS and 5-year CSS compared to those aged≥65 years.Patients with urothelial carcinoma showed better 5-year OS than those with non-urothelial carcinoma.Additionally,tumor stage,tumor grade,and LNM were negatively associated with 5-year OS and 5-year CSS.On multivariate analysis,only tumor grade and LNM remained statistically significant(P<0.05).Conclusions:Tumor grade and LNM were identified as independent prognostic risk factors for 5-year OS and 5-year CSS following RC combined with Mainz PouchⅡ bladder reconstruction.Moreover,the application of RC combined with Mainz Pouch Ⅱ bladder reconstruction should consider the patient's preferences and physical condition.

Objective:To investigate the association between exposures of seven classic phthalate metabolites and fertility among preconception women before pregnancy.Methods:Based on the preconception cohort, a nested case-control study was carried out. Preconception couples who were recruited from Shanghai Jiading District Maternal and Child Health Hospital from 2016 to 2020 with intention to conception were selected. A total of 100 women whose time to pregnancy (TTP) >12 months were selected as the case group, and 151 women with the TTP≤12 months were randomly selected as control group. The single and joint effects on fertility of phthalate metabolites were analyzed by the comparison between the two groups via the multivariate binary logistic regression model, and the weighted quantile sum regression model.Results:There were significant differences in mono-(2-ethylhexyl) phthalate (MEHP), mono-(2-carboxmethyl)-hexyl phthalate (MCMHP), the sum of the detected concentrations of MEHP, MCMHP, mono-(2-ethyl-5-oxohexyl) phthalate, mono-(2-ethyl-5-hydroxyhexyl) phthalate and mono-(2-ethyl-5-carboxypentyl) phthalate (∑DEHP), and the sum of the detected concentrations of the seven phthalate metabolites (∑PAE) exposure levels between the case group and control group ( P=0.005, P<0.001, P=0.001, P=0.007). As shown by the multivariate binary logistic regression model, women with MEHP, MCMHP, ∑DEHP, ∑PAE at the highest quartile level were found to have significantly increased risk of infertility compared with women at the lowest quartile level (a OR=2.40, 95% CI: 1.12-5.15, P=0.025; a OR=7.74, 95% CI: 3.14-19.08, P<0.001; a OR=4.35, 95% CI: 1.93-9.79, P<0.001; a OR=2.04, 95% CI: 0.96-4.34, P=0.044). The weighted quantile sum regression model demonstrated that the exposure to seven phthalate metabolite mixtures in preconception was associated with an increased risk of infertility ( P<0.05). Among the seven phthalate metabolites, MCMHP had the largest contribution weight (61.7%). Conclusion:Women of childbearing age should avoid the exposure to phthalates in the environment when preparing for conception, such as to reduce the consumption of highly processed foods and foods packaged with plastic/polyvinyl chloride materials to promote fertility.

Objective:To investigate the association between exposures of seven classic phthalate metabolites and fertility among preconception women before pregnancy.Methods:Based on the preconception cohort, a nested case-control study was carried out. Preconception couples who were recruited from Shanghai Jiading District Maternal and Child Health Hospital from 2016 to 2020 with intention to conception were selected. A total of 100 women whose time to pregnancy (TTP) >12 months were selected as the case group, and 151 women with the TTP≤12 months were randomly selected as control group. The single and joint effects on fertility of phthalate metabolites were analyzed by the comparison between the two groups via the multivariate binary logistic regression model, and the weighted quantile sum regression model.Results:There were significant differences in mono-(2-ethylhexyl) phthalate (MEHP), mono-(2-carboxmethyl)-hexyl phthalate (MCMHP), the sum of the detected concentrations of MEHP, MCMHP, mono-(2-ethyl-5-oxohexyl) phthalate, mono-(2-ethyl-5-hydroxyhexyl) phthalate and mono-(2-ethyl-5-carboxypentyl) phthalate (∑DEHP), and the sum of the detected concentrations of the seven phthalate metabolites (∑PAE) exposure levels between the case group and control group ( P=0.005, P<0.001, P=0.001, P=0.007). As shown by the multivariate binary logistic regression model, women with MEHP, MCMHP, ∑DEHP, ∑PAE at the highest quartile level were found to have significantly increased risk of infertility compared with women at the lowest quartile level (a OR=2.40, 95% CI: 1.12-5.15, P=0.025; a OR=7.74, 95% CI: 3.14-19.08, P<0.001; a OR=4.35, 95% CI: 1.93-9.79, P<0.001; a OR=2.04, 95% CI: 0.96-4.34, P=0.044). The weighted quantile sum regression model demonstrated that the exposure to seven phthalate metabolite mixtures in preconception was associated with an increased risk of infertility ( P<0.05). Among the seven phthalate metabolites, MCMHP had the largest contribution weight (61.7%). Conclusion:Women of childbearing age should avoid the exposure to phthalates in the environment when preparing for conception, such as to reduce the consumption of highly processed foods and foods packaged with plastic/polyvinyl chloride materials to promote fertility.

Objective:To investigate the attitude of neonatologists toward the treatment of extremely preterm infants (EPIs) in China.Methods:A cross-sectional survey was conducted using a questionnaire designed and posted on Wenjuanxing, a web-based survey platform, from June to July 2021. The respondents were neonatal physicians in various provinces and cities in China. The questionnaire covered the basic information, treatment experience and attitude towards EPIs, and opinions on the current definition of the preterm infant in China. The results were described or analyzed using the Chi-square test.Results:A total of 1 066 valid replies were collected. The respondents included 322 males and 744 females, among whom 78.1% (832/1 066) were assistant director physicians or director physicians, 82.8% (882/1 066) were from tertiary hospitals, and 83.0% (885/1 066) had the experiences of treating EPIs. In terms of the attitude toward the treatment of EPIs, 63.0% (672/1 066) of the respondents suggested that the lower limit of gestational age for EPIs requiring active resuscitation should be defined at 25 gestational weeks or less. Moreover, 57.1% (609/1 066) considered that the current domestic definition of preterm infants as 28 gestational age or above was inappropriate, and 75.2% (458/609) considered that the lower limit defined as preterm infants should be 25 gestational weeks or less. Concerning the treatment experience in EPIs, 54.3% (579/1 066) of the respondents suggested that in their hospital, withdrawing treatment in EPIs was common or very common, and 83.3% (888/1 066) considered that the main reason for withdrawing treatment was family members' concerns about the prognosis. Those who hesitated about treating the EPIs accounted for 71.6% (763/1 066), and 83.9% (640/763) hesitated due to the poor prognosis and possible medical disputes. Moreover, 32.7% (349/1 066) of the respondents or their colleagues had been involved in medical disputes about whether to treat EPIs, and 74.8% (797/1 066) believed that the patients should be the decision-maker on whether to treat EPIs or not.Conclusion:Most neonatal physicians in this survey hold a positive attitude toward the treatment of EPIs and believe that the lower limit of gestational age for preterm infants should be lowered. However, a hesitating attitude to the care of EPIs is still common, and uncertainty about the prognosis of EPIs remains a concern.