Robot-assisted nephroureterectomy: surgical and mid-term oncological outcomes in over 1100 patients (ROBUUST 2.0 collaborative group).

OBJECTIVE: To analyse surgical, functional, and mid-term oncological outcomes of robot-assisted nephroureterectomy (RANU) in a contemporary large multi-institutional setting. PATIENTS AND METHODS: Data were retrieved from the ROBotic surgery for Upper tract Urothelial cancer STtudy (ROBUUST) 2.0 database, an international, multicentre registry encompassing data of patients with upper urinary tract urothelial carcinoma undergoing curative surgery between 2015 and 2022. The analysis included all consecutive patients undergoing RANU except those with missing data in predictors. Detailed surgical, pathological, and postoperative functional data were recorded and analysed. Oncological time-to-event outcomes were: recurrence-free survival (RFS), metastasis-free survival (MFS), cancer-specific survival (CSS), and overall survival (OS). Survival analysis was performed using the Kaplan-Meier method, with a 3-year cut-off. A multivariable Cox proportional hazard model was built to evaluate predictors of each oncological outcome. RESULTS: A total of 1118 patients underwent RANU during the study period. The postoperative complications rate was 14.1%; the positive surgical margin rate was 4.7%. A postoperative median (interquartile range) estimated glomerular filtration rate decrease of -13.1 (-27.5 to 0) mL/min/1.73 m2 from baseline was observed. The 3-year RFS was 59% and the 3-year MFS was 76%, with a 3-year OS and CSS of 76% and 88%, respectively. Significant predictors of worse oncological outcomes were bladder-cuff excision, high-grade tumour, pathological T stage ≥3, and nodal involvement. CONCLUSIONS: The present study contributes to the growing body of evidence supporting the increasing adoption of RANU. The procedure consistently offers low surgical morbidity and can provide favourable mid-term oncological outcomes, mirroring those of open NU, even in non-organ-confined disease.

Role of neoadjuvant chemotherapy in patients with locally advanced and clinically positive nodes Upper Tract Urothelial Carcinoma treated with Nephroureterectomy: real-world data from the ROBUUST 2.0 Registry.

PURPOSE: To assess the impact of neoadjuvant and adjuvant chemotherapy on survival outcomes, within a large multicenter cohort of Upper tract urothelial carcinoma patients treated with Nephroureterectomy. METHODS: A multicenter retrospective analysis utilizing the Robotic surgery for Upper Tract Urothelial Cancer Study registry was performed. Baseline, preoperative, perioperative, and pathologic variables of three groups of patients receiving surgery only, neoadjuvant or adjuvant chemotherapy were compared. Categorical and continuous variables among the three subgroups were compared with Chi square and ANOVA tests. The impact of perioperative chemotherapy on survival outcomes was assessed with the Kaplan Meier method. Univariable and multivariable Cox regression analyses were performed to identify predictors of survival. RESULTS: Overall, 1,994 patients were included. Overall and Clavien grade ≥3 complications rates were comparable among the three subgroups (p = 0.65 and p = 0.92). At Kaplan Meier analysis, neoadjuvant chemotherapy significantly improved cancer-specific survival (p = 0.03) and overall survival (p = 0.03) probabilities of patients with cT ≥ 3 tumors and of those with positive cN (p = 0.03 and p = 0.02). On multivariable analysis, neoadjuvant chemotherapy was independently associated with an improvement of cancer-specific survival in cT ≥ 3 patients (HR 0.44; p = 0.04), and of both cancer-specific survival (HR 0.50; p = 0.03) and overall survival (HR 0.53; p = 0.02) probabilities in positive cN patients. CONCLUSIONS: This large multicenter retrospective analysis suggests significant survival benefit in Upper tract urothelial carcinoma patients with either locally advanced or clinically positive nodes disease receiving neoadjuvant chemotherapy. These findings can be regarded as "hypothesis generating", stimulating future trials focusing on such advanced stages.

Study of Raman scattering enhancement method based on optical multiplexing for on-line detection of gas components in strong-impact environments.

On-line gas detection under strong impact such as combustion and explosion is of great significance for understanding the reaction processes. To realize simultaneous on-line detection of various gases under strong impact, an approach based on optical multiplexing for enhancing spontaneous Raman scattering is proposed. A single beam is transmitted several times using optical fibers through a specific measurement point in the reaction zone. Thus, the excitation light intensity at the measurement point is enhanced and the Raman signal intensity is substantially increased. Indeed, the signal intensity can be increased by a factor of ∼10, and the constituent gases in air can be detected with sub-second time resolution, under a 100 g impact.

Outcomes of Lymph Node Dissection in Nephroureterectomy in the Treatment of Upper Tract Urothelial Carcinoma: Analysis of the ROBUUST Registry.

PURPOSE: We sought to evaluate outcomes of lymph node dissection (LND) in patients with upper tract urothelial carcinoma. MATERIALS AND METHODS: We performed a multicenter retrospective analysis utilizing the ROBUUST (for RObotic surgery for Upper Tract Urothelial Cancer Study) registry for patients who did not undergo LND (pNx), LND with negative lymph nodes (pN0) and LND with positive nodes (pN+). Primary and secondary outcomes were overall survival (OS) and recurrence-free survival (RFS). Multivariable analyses evaluated predictors of outcomes and pathological node positivity. Kaplan-Meier analyses (KMAs) compared survival outcomes. RESULTS: A total of 877 patients were analyzed (LND performed in 358 [40.8%]/pN+ in 73 [8.3%]). Median nodes obtained were 10.2 for pN+ and 9.8 for pN0. Multivariable analyses noted increasing age (OR 1.1, p <0.001), pN+ (OR 3.1, p <0.001) and pathological stage pTis/3/4 (OR 3.4, p <0.001) as predictors for all-cause mortality. Clinical high-grade tumors (OR 11.74, p=0.015) and increasing tumor size (OR 1.14, p=0.001) were predictive for lymph node positivity. KMAs for pNx, pN0 and pN+ demonstrated 2-year OS of 80%, 86% and 42% (p <0.001) and 2-year RFS of 53%, 61% and 35% (p <0.001), respectively. KMAs comparing pNx, pN0 ≥10 nodes and pN0 <10 nodes showed no significant difference in 2-year OS (82% vs 85% vs 84%, p=0.6) but elicited significantly higher 2-year RFS in the pN0 ≥10 group (60% vs 74% vs 54%, p=0.043). CONCLUSIONS: LND during nephroureterectomy in patients with positive lymph nodes provides prognostic data, but is not associated with improved OS. LND yields ≥10 in patients with clinical node negative disease were associated with improved RFS. In high-grade and large tumors, lymphadenectomy should be considered.

Receptor activity-modifying protein 1 regulates mouse skin fibroblast proliferation via the Gαi3-PKA-CREB-YAP axis.

BACKGROUND: Skin innervation is crucial for normal wound healing. However, the relationship between nerve receptors and wound healing and the intrinsic mechanism remains to be further identified. In this study, we investigated the role of a calcitonin gene-related peptide (CGRP) receptor component, receptor activity-modifying protein 1 (RAMP1), in mouse skin fibroblast (MSF) proliferation. METHODS: In vivo, Western blotting and immunohistochemical (IHC) staining of mouse skin wounds tissue was used to detect changes in RAMP1 expression. In vitro, RAMP1 was overexpressed in MSF cell lines by infection with Tet-On-Flag-RAMP1 lentivirus and doxycycline (DOX) induction. An IncuCyte S3 Live-Cell Analysis System was used to assess and compare the proliferation rate differences between different treatment groups. Total protein and subcellular extraction Western blot analysis, quantitative real-time-polymerase chain reaction (qPCR) analysis, and immunofluorescence (IF) staining analysis were conducted to detect signalling molecule expression and/or distribution. The CUT & RUN assay and dual-luciferase reporter assay were applied to measure protein-DNA interactions. RESULTS: RAMP1 expression levels were altered during skin wound healing in mice. RAMP1 overexpression promoted MSF proliferation. Mechanistically, total Yes-associated protein (YAP) and nuclear YAP protein expression was increased in RAMP1-overexpressing MSFs. RAMP1 overexpression increased inhibitory guanine nucleotide-binding protein (G protein) α subunit 3 (Gαi3) expression and activated downstream protein kinase A (PKA), and both elevated the expression of cyclic adenosine monophosphate (cAMP) response element-binding protein (CREB) and activated it, promoting the transcription of YAP, elevating the total YAP level and promoting MSF proliferation. CONCLUSIONS: Based on these data, we report, for the first time, that changes in the total RAMP1 levels during wound healing and RAMP1 overexpression alone can promote MSF proliferation via the Gαi3-PKA-CREB-YAP axis, a finding critical for understanding RAMP1 function, suggesting that this pathway is an attractive and accurate nerve target for skin wound treatment. Video Abstract.

Optical weak measurement for the precise thickness determination of an ultra-thin film.

A total internal reflection system based on the weak value amplification principle is set up for the precise measurement of the thickness of an ultra-thin film. In this system, the film thickness is derived from the change of the double-peak pointer caused by the effective refractive index of the film, which is correlated to its thickness. The sensitivity and resolution of this system reached 2727.21 nm/RIU and 7.2×10-6 R I U, respectively, determined by using a sodium chloride solution with a refractive index of 1.331911. The growth process of TA/Fe(III) assembled films with thicknesses in the few nanometers range is monitored using the as-set-up system, and the experimental results are consistent with a theoretical calculation based on the Maxwell Garnett effective medium. Additionally, we theoretically calculated the detection limit for the thickness measurement of the film as 22 pm. We clearly provide a potential method for the precise measurement of the thickness of an ultra-thin film.

Risk Factors for Intravesical Recurrence after Minimally Invasive Nephroureterectomy for Upper Tract Urothelial Cancer (ROBUUST Collaboration).

PURPOSE: Intravesical recurrence (IVR) after radical nephroureterectomy (RNU) for upper tract urothelial carcinoma (UTUC) has an incidence of approximately 20%-50%. Studies to date have been composed of mixed treatment cohorts-open, laparoscopic and robotic. The objective of this study is to assess clinicopathological risk factors for intravesical recurrence after RNU for UTUC in a completely minimally invasive cohort. MATERIALS AND METHODS: We performed a multicenter, retrospective analysis of 485 patients with UTUC without prior or concurrent bladder cancer who underwent robotic or laparoscopic RNU. Patients were selected from an international cohort of 17 institutions across the United States, Europe and Asia. Univariate and multiple Cox regression models were used to identify risk factors for bladder recurrence. RESULTS: A total of 485 (396 robotic, 89 laparoscopic) patients were included in analysis. Overall, 110 (22.7%) of patients developed IVR. The average time to recurrence was 15.2 months (SD 15.5 months). Hypertension was a significant risk factor on multiple regression (HR 1.99, CI 1.06; 3.71, p=0.030). Diagnostic ureteroscopic biopsy incurred a 50% higher chance of developing IVR (HR 1.49, CI 1.00; 2.20, p=0.048). Treatment specific risk factors included positive surgical margins (HR 3.36, CI 1.36; 8.33, p=0.009) and transurethral resection for bladder cuff management (HR 2.73, CI 1.10; 6.76, p=0.031). CONCLUSIONS: IVR after minimally invasive RNU for UTUC is a relatively common event. Risk factors include a ureteroscopic biopsy, transurethral resection of the bladder cuff, and positive surgical margins. When possible, avoidance of transurethral resection of the bladder cuff and alternative strategies for obtaining biopsy tissue sample should be considered.

Clinical outcomes of 168 Chinese patients after local surgery for bone metastases arising from advanced renal cell carcinoma.

BACKGROUND: The effectiveness of local surgery for bone metastases (BM) arising from renal cell carcinoma (RCC) remains uncertain. Herein, the authors performed what to the best of their knowledge is the first investigation of the outcomes of Chinese patients with RCC and BM. METHODS: Data were collected for 168 patients with RCC and BM who were treated at Changzheng Hospital in Shanghai, China, between March 2009 and December 2018. All patients underwent local surgery for BM arising from RCC. Overall survival (OS) was defined as the interval between the date of local surgery and death or last follow-up and was estimated using the Kaplan-Meier method. Univariate and multivariable Cox proportional hazards analyses were used to identify significant prognostic factors. RESULTS: The median OS in the study cohort was 43 months (range, 0-113 months). The 1-year, 3-year, and 5-year survival rates after surgery were 77.4%, 55.9%, and 31.8%, respectively. Univariate analyses demonstrated significant survival differences associated with sex (P = .003), the number of preoperative metastatic sites (BM alone or BM with concomitant metastasis; P < .001), and the number of BM (single or multiple; P = .008). OS also did not appear to differ regardless of whether the patients received postsurgical targeted therapy. Multivariable Cox regression demonstrated that the following characteristics were independent predictors of OS: the number of preoperative metastatic sites, International Metastatic Renal Cell Carcinoma Database Consortium risk score, and Memorial Sloan Kettering Cancer Center score. CONCLUSIONS: Careful patient selection for local surgery is of paramount importance. The prognoses of patients in the Memorial Sloan Kettering Cancer Center-based and International Metastatic Renal Cell Carcinoma Database Consortium-based poor-risk groups were much worse than those of the intermediate-risk groups. In current clinical practice, "stratified treatment" can be performed according to these criteria.

Effects of breathing exercises using home-based positive pressure in the expiratory phase in patients with COPD.

BACKGROUND: Patients with chronic obstructive pulmonary disease (COPD) commonly have higher intrinsic positive end-expiratory pressure (PEEPi). A breathing exercise programme strategy employing an appropriate PEEP may improve their pulmonary functional capacity, exercise tolerance and health-related quality of life. Breathing with an expiratory resistive load, which is a method of modulating spontaneous breathing against PEEPi, has not been fully studied in patients with COPD. The objective of this study was to investigate the role of changing spontaneous breathing in home-based conditions and regulating spontaneous breathing with breathing exercises in patients with COPD. METHODS: This was a prospective randomised trial including 64 patients with a diagnosis of stage III or IV COPD. Patients were randomised into two groups: standard treatment and standard treatment combined with breathing exercise rehabilitation. The effects of the treatments on the COPD assessment test (CAT) score, 6-minute walk test (6MWT) results and pulmonary function were compared at 0, 6, 12 and 18 months within and between the two groups. RESULTS: All outcomes showed no significant differences between the two groups at the beginning of the study, while the 6MWT and CAT scores exhibited clinically and statistically significant improvements (p<0.001) by the end of the study. At month 18, the change in the predicted percentage of forced expiratory volume in 1 s (FEV1%pred) differed between the two groups (p<0.05). In addition, there were statistically significant differences in the 6MWT results, CAT scores and FEV1%pred values between the baseline and month 18 (p<0.0001) in the intervention group. CONCLUSIONS: Improvements in 6MWT results, pulmonary function and CAT scores are associated with a successful response to breathing against PEEPi in patients with COPD. TRIAL REGISTRATION: This trial was registered at research registry.com (identifier research registry 4816).

n-Butyllithium Catalyzed Selective Hydroboration of Aldehydes and Ketones.

Highly efficient and selective hydroboration of aldehydes and ketones with HBpin is achieved by using the simple and convenient n-BuLi as a catalyst. The reaction proceeds rapidly with low catalyst loading (0.1-0.5 mol %) under mild conditions. Key features include the high catalytic efficiency, exceptional functional group compatibility, ample substrate scope, and high selectivity for aldehydes over ketones. Computational studies were carried out to provide a mechanistic insight into the n-BuLi catalyzed hydroboration of aldehydes/ketones with HBpin.

Digital quantification of DNA by mapping polarization degree related with coding gold nanorods.

The development of highly sensitive and low-cost methods for detecting DNA is of critical importance. Here, we describe a strategy for the highly sensitive and low-cost digital detection of target DNA. Individual DNA molecules were encoded with a single gold nanorod (Au NR), which was then separated and enriched using the magnetic immune-separation process, followed by dehybridization and dispersion into a buffer solution and immobilization on glass slides for polarized dark-field microscopic imaging. With the imaging we can get the first three data sets of the Stokes vector, and the experimental degree of the linear polarization of the light scattered by the Au NR was obtained. Using the Monte Carlo simulation program, the Muller matrix of the Au NRs was simulated and the simulated degree of the linear polarization was calculated to be 0.58. Based on the experimental and simulated degree of the linear polarization, the Au NRs were identified and quantified with an in-house Matlab program, and the concentration of the target DNA at the femtomolar level was therefore achieved.

Digital Concentration Readout of DNA by Absolute Quantification of Optically Countable Gold Nanorods.

Isolation and amplification procedures are indispensable for traditional single-molecule detection methods applied in low-abundance biomolecule analysis. Here, we describe a method for ultrasensitive detection of DNA through encoding a single target molecule with a single-countable nanometer-sized substitute (gold nanorod, AuNR) and enrichment of the substitute AuNRs into a limited region followed by accurate microscopic enumeration. The enrichment and the bright distinct color allow the AuNRs to be efficiently counted from as few as 1-2 to tens of thousands in 3 µL of test solution, which demonstrates the ability of rapid digital concentration readout of the target DNA. On this basis, a detection limit of 6.5 aM was achieved for DNA associated with human papillomavirus (HPV). Notably, our method requires neither complicated isolation and amplification procedures nor extremely expensive instruments and consumables.

WT1 is involved in the Akt-JNK pathway dependent autophagy through directly regulating Gas1 expression in human osteosarcoma cells.

Macroautophagy (herein termed autophagy) works as a protective mechanism in tumorigenesis and development under metabolic stress condition. Multitudes of genes have been found involved in this process during past decades. In the present study, we report that Wilm's tumor suppressor1 (WT1) is involved in autophagy in osteosarcoma (OS) cells. WT1, a transcription factor with multitude of target genes, expresses in a majority of cancer types. Though wide-ranging effect of WT1 is now well documented, the function of WT1 in tumors remains poorly defined. In this chapter, it is found that high expression of WT1 positively correlates with active autophagy in human osteosarcoma cells. And further study on cell signaling pathway illustrates that Akt/JNK pathway acts as a positive regulator of autophagy induced by WT1. Here, we present evidence that WT1 modulates Akt/JNK signaling pathway mediated autophagy by controlling the expression of growth arrest-specific 1 (Gas1). We show that WT1 is required for Gas1 transcription in osteosarcoma cells. And Gas1 is upregulated followed WT1 overexpression in a time-dependent manner. Loss of Gas1 results in a reduction of WT1-induced autophagy.

Interleukin 10-expressing B cells inhibit tumor-infiltrating T cell function and correlate with T cell Tim-3 expression in renal cell carcinoma.

Renal cell carcinoma is among the leading causes of cancer-related death and was found to induce IL-10. We started by focusing on IL-10-secreting cells in tumor-infiltrating lymphocytes in renal cell carcinoma patients and observed that both CD3(+) T cells and CD19(+) B cells contributed to an elevated IL-10 expression. We then focused on IL-10-expressing B cells, and found that compared to non-IL-10-producing B cells, the IL-10-expressing B cells had significantly lower levels of CD19 and CD20 expression, a lack of IgM and IgD expression, while the level of CD27 was elevated. Moreover, culturing under unstimulated conditions resulted in higher antibody production by these IL-10-producing B cells than their peripheral blood counterparts, which strongly suggested that they are plasmablast-differentiating cells. Both IgA and IgG subtypes were found but IgA had a higher relative abundance in the tumor-infiltrating fraction. We then observed inverse correlations between the frequency of IL-10-producing B cells and pro-inflammatory cytokine-producing T cells and T cell proliferation. The expression of T cell exhaustion marker Tim-3, however, was upregulated in patients with high frequencies of IL-10-producing B cells. Moreover, supernatant from tumor B cells suppressed T cell inflammation. In addition, frequencies of IL-10-producing tumor-infiltrating B cells were inversely correlated with resected tumor size, and were higher in later stage tumors. Together, our data demonstrated that IL-10-producing B cells had plasmablast-differentiating phenotype, and could contribute to T cell immunosuppression in renal cell carcinoma.

Tim-3 expression represents dysfunctional tumor infiltrating T cells in renal cell carcinoma.

PURPOSE: Renal cell carcinoma (RCC) is the most common cancer of kidney. Evidences have shown that RCC is sensitive to various immunotherapies. Tim-3 plays a role in suppressing Th1-mediated immune responses. However, no study has yet examined the effect of Tim-3 on tumor infiltrating lymphocytes (TILs) in RCC. METHODS: We investigated the expression and function of Tim-3 on TIL CD4+ T cells and TIL CD8+ T cells from 30 RCC patients. RESULTS: Levels of Tim-3 were significantly increased on both TIL CD4+ T cells and TIL CD8+ T cells and were associated with higher stages of the cancer. Also, GATA-3 and interferon gamma (IFN-γ) were down-regulated, whereas T-bet was up-regulated in TIL Tim-3+ T cells, indicating that Tim-3 expression defined a population of dysfunctional TIL Th1/Tc1 cells. Mechanism analyses showed that TIL Tim-3-expressing CD8+ T cells exhibited impaired Stat5 and p38 signaling pathway. Blocking the Tim-3 pathway restored cell proliferation and increased IFN-γ production in TIL CD4+ and CD8+ T cells of RCC. CONCLUSIONS: These results suggest that Tim-3 may be used as a novel target for increasing immune responses in RCC tumor microenvironment.

Microendoscopic discectomy versus open discectomy for lumbar disc herniation: a meta-analysis.

PURPOSE: To compare the outcomes of microendoscopic discectomy and open discectomy for patients with lumbar disc herniation. METHODS: An extensive search of studies was performed in PubMed, Medline, Embase, Cochrane library and Google Scholar. The following outcome measures were extracted: visual analogue scale (VAS), Oswestry disability index (ODI), complication, operation time, blood loss and length of hospital stay. Data analysis was conducted with RevMan 5.0. RESULTS: Five randomized controlled trials involving 501 patients were included in this meta-analysis. The pooled analysis showed that there was no significant difference in the VAS, ODI or complication between the two groups. However, compared with the open discectomy, the microendoscopic discectomy was associated with less blood loss [WMD = -151.01 (-288.22, -13.80), P = 0.03], shorter length of hospital stay [WMD = -69.33 (-110.39, -28.28), P = 0.0009], and longer operation time [WMD = 18.80 (7.83, 29.76), P = 0.0008]. CONCLUSIONS: Microendoscopic discectomy, which requires a demanding learning curve, may be a safe and effective alternative to conventional open discectomy for patients with lumbar disc herniation.

Propensity-score matched analysis comparing robot-assisted with laparoscopic partial nephrectomy.

OBJECTIVES: To compare the peri-operative and early renal functional outcomes of robot-assisted partial nephrectomy (RAPN) and laparoscopic partial nephrectomy (LPN) for kidney tumours. MATERIALS AND METHODS: A total of 237 patients fulfilling the selection criteria were included, of whom 146 and 91 patients were treated with LPN and RAPN, respectively. To adjust for potential baseline confounders, propensity-score matching was performed. A favourable outcome was defined as a warm ischaemia time (WIT) of ≤20 min, negative surgical margins, no surgical conversion, no Clavien ≥3 complications and no postoperative chronic kidney disease (CKD) upstaging. Descriptive statistics and multivariable logistic regression analyses were performed before and after propensity-score matching. RESULTS: Within the propensity-score-matched cohort, the RAPN group was associated with significantly lower estimated blood loss (EBL; 156 vs 198 mL, mean difference [MD] = -42; P = 0.025), a shorter WIT (22.8 vs 31 min, MD = -8.2; P < 0.001) and a higher proportion of malignant lesions (88.4 vs 67.5%; odds ratio [OR]: 2.6; 95% confidence interval [CI]: 1.2-5.67; P = 0.023). With regard to early renal functional outcomes, the mean last estimated glomerular filtration rate was 95.8 and 89.4 mL/min per 1.73 m(2) (MD = 6.4; P = 0.01), with a mean ± sd percentage change of -4.8 ± 17.9 and -12.2 ± 16.6 (MD = 7.4; P = 0.018) in the RAPN and LPN groups, respectively. The intra-operative complication rate was significantly lower in the RAPN group (1.3 vs 11.7%; OR 0.1, 95% CI 0.01-0.81; P = 0.018). On multivariable analysis, surgical approach (RAPN vs LPN, OR 5.457, 95% CI 2.075-14.346; P = 0.001), Charlson Comorbidity Index (OR 0.223; 95% CI 0.062-0.811; P = 0.023), diameter-axial-polar score (OR 0.488, 95% CI 0.329-0.723; P < 0.001) and preoperative CKD stage (OR 3.189, 95% CI 1.204-8.446; P = 0.020) were found to be independent predictors of obtaining a favourable outcome. CONCLUSIONS: After adjusting for potential treatment selection biases, RAPN was found to be superior to LPN for peri-operative outcomes (EBL, WIT and intra-operative complications) and early renal functional preservation.