Stage Group Research Articles

Florfenicol can inhibit chick growth and lead to immunosuppression1

Florfenicol (FLO) is a chemically synthesized broad-spectrum antimicrobial agent of amide alcohols for animals, which is one of the most widely used antimicrobials in livestock, poultry, and aquaculture. With the use of FLO, more and more attention has been paid to its hematopoietic toxicity, immunotoxicity, genotoxicity, and embryotoxicity. In this study, SPF chicks at the age of 3 d began to drink water with the FLO at a dose of 100 mg L-1 for six consecutive days, and the growth performance of chicks was monitored, the effect of FLO on immune organs was detected by pathological examination and TUNEL apoptosis staining. In order to evaluate the level of organism immunity, the level of Newcastle disease virus antibody in serum was detected by hemagglutination inhibition test, the content of cytokines (IL-1, IL-2, IL-6, TNF-α, IFN-γ) in serum was detected by ELISA, and the transcription of interferon-related genes (IRF-7, 2’-5’OAS, Mx1) and cytokine genes (IL-6, TNF-α, IFN-γ) in immune organs were detected by real time fluorescence quantitative PCR. The results showed that the early application of FLO could inhibit the growth and development of chicks, and the body weight and immune organ index of the treatment group were lower than those of the control group. Histopathological examination showed that there was a decrease in the number of lymphocytes in the bursa of Fabricius in the treatment group in the early stage of drug withdrawal, and the results of TUNEL apoptosis staining in the bursa of Fabricius showed that obvious lymphocyte apoptosis occurred in the FLO treatment group. Compared with the control group, the transcription levels of interferon-related genes IRF-7, 2’-5’OAS, and cytokine genes IL-6, TNF-α and IFN-γ in FLO treatment group decreased to a certain extent, while the transcription level of Mx1 gene had no significant difference at all time points. The level of serum NDV antibody and the contents of cytokines IL-1, IL-2 and IFN-γ in the FLO treatment group were significantly lower than those in the control group in the early stage of drug withdrawal, but recovered gradually in the later stage. This study showed that FLO has a certain degree of effect on the immune function of chicks, and the results of the study laid the foundation for further research on the mechanism of FLO-induced immunotoxicity.

Hepatocyte Growth Factor Delivered by Nanocomposites for Gene Therapy of Bleomycin-Induced Pulmonary Fibrosis in Rats.

Pulmonary fibrosis (PF) is a chronic and progressive interstitial lung disease. There is no effective treatment for PF. Hepatocyte growth factor (HGF) has anti-inflammatory and antifibrotic effects but has limited potential owing to its short half-life. To increase the transfection efficiency of pVAX-HGF, we prepared polyethyleneiminepolyethylene glycol: polyethyleneimine/pVAX-HGF (PEG-PEI: PEI/pVAX-HGF) nanocomposite loaded with a plasmid encoding the HGF gene. The PEG-PEI:PEI/pVAX-HGF characteristics, including morphology, particle size, zeta-potential, and DNA entrapment efficiency, were investigated. The pVAX-HGF nanocomposites with low toxicity and high transfection efficiency were screened by cell viability assay and cell transfection. The antifibrotic effect of pVAX-HGF nanocomposite on PF rats induced by bleomycin (BLM) was evaluated by pulmonary function measurement, pathological examination and collagen content assay. Different nanocomposites were prepared to deliver pVAX-HGF, in which mix1 (PEGPEI: PEI/pVAX-HGF) has lower potential and better entrapment ability. PEI/pVAX-HGF (N/P=25) nanocomposite with low toxicity and high transfection efficiency was administered to PF rats. After treatment with mix 1/pVAX-HGF, the index of lung function(including EF50, MV, TV, PEF and PIF) in mix 1/pVAX-HGF group was higher than that of the PF group. The number of cells in BALF of the mix 1/pVAX-HGF group was significantly lower than that of the PF groups, and the content of hydroxyproline(HYP) and collagen Type I (Col-I) in the lung of the mix 1/pVAX-HGF group was much lower than that of the PF groups in the early stage. The result of pathological examination showed that rats in the mix1/pVAX-HGF group showed obviously reduced alveolar septal thickening, fewer infiltrated inflammatory cells and less collagen deposition. The PEG-PEI:PEI/pVAX-HGF nanocomposite can ameliorate PF induced by BLM. The pVAX-HGF nanocomposite is a latent therapeutic strategy for PF.

MiRNA Expression Profiles in an Ectopic Endometrium of Patients at Different Stage of Endometriosis

Endometriosis is a debilitating disease that affects up to 15% of women worldwide. Delayed diagnosis, lack of noninvasive biomarker and unexplained pathogenesis are key problems for specialists and patients alike. MiRNAs are a class of noncoding RNAs that regulate myriad of cellular functions. These gene expression regulators may prove to be attractive biomarkers with diagnostic and prognostic value for endometriosis.
 Tissue samples were collected from the participants enrolled in the study during laparoscopy (patients) and hysteroscopy (control group). After RNA isolation (miRNeasy MiniKit, Qiagen) based on the disease stage, three pools – each containing 15 RNA samples, were constructed: 1) early stage endometriosis, 2) late stage endometriosis, and 3) healthy controls. Each pool sample was subjected to reverse transcription via miScript II RT Kit, Qiagen to obtain cDNA. SYBR Green based Real-time PCR assay was used to determine the expression profile of 84 miRNAs (Human miFinder miRNA PCR Array, Qiagen).
 We detected 32 differently expressed miRNAs between early stage endometriosis and control group, and 51 differently expressed miRNAs between advanced stage endometriosis and control group. Three miRNAs were differentially expressed with more than 10-fold change in the early stage group and thn miRNAs showed more than 10-fold change in the advanced stages group. The three miRNAs with the highest fold change in the expression levels in both case groups were nominated as potential biomarkers for endometriosis. The results of the analysis of their target genes supports the role of deregulation of apoptosis, angiogenesis, epithelial-mesenchymal transition, and various other cellular signalling pathways in endometriosis development.

Effects of different conditions on the artificial incubation effect and physiological indexes of redclaw crayfish eggs

We explored the effects of different conditions on the artificial incubation of redclaw crayfish eggs in an effort to improve this process. Samples at the egg and juvenile stages were selected. The samples at different stages were separated from the pleopods, then they were placed in incubator boxes and sterilized with different disinfectant solutions. The density was 300,400 and 500 eggs/incubator box, the vibration frequency was 11,16 and 26 vibrations/min, and the water circulation cycle was 2.1, 4.8 and 7.1 cycles/h. The results showed the eggs disinfected with 3000 ppm formaldehyde for 15 min had stronger antioxidant capacity. The hatching and survival rates of five pairs of appendage stage group were significantly lower than those of other groups. In the egg stage, acid phosphatase (ACP) level of compound eye pigmentation stage group was significantly higher than those of other groups. In the juvenile stage, malondialdehyde (MDA) content of five pairs of appendage stage group was significantly higher than those of other groups. The survival rate of 500 eggs/box group was significantly higher than that of other groups. In the egg stage, alkaline phosphatase (AKP) level of 400 eggs/box group was significantly higher than that of other groups. The survival rate of 11 vibrations/min group was significantly higher than that of other groups. In the egg stage, ACP and AKP levels of 11 vibrations/min group were significantly higher than those of 26 vibrations/min group. In the juvenile stage, superoxide dismutase (SOD), ACP and AKP levels of 11 vibrations/min group was significantly higher than those of 26 vibrations/min group. In the juvenile stage, AKP level of 4.8 cycles/h group was significantly lower than that of other groups. In conclusion, egg development at the stage after seven pairs of appendages, with a density of 400 eggs/box, vibration frequencies set at 11 vibrations/min achieved high hatching rates (93.58 %) and survival rates (75.67 %). Moreover, bronopol or hydrogen peroxide might have a better choice to replace formaldehyde if further exploration was conducted to reduce stimulation of the in vitro-grown egg. These conditions could be used on a large scale to optimize the production of redclaw crayfish.

Correlation of Perfusion Metrics with Ki-67 Proliferation Index and Axillary Involvement as a Prognostic Marker in Breast Carcinoma Cases: A Dynamic Contrast-Enhanced Perfusion MRI Study.

Our study aims to reveal clinically helpful prognostic markers using quantitative radiologic data from perfusion magnetic resonance imaging for patients with locally advanced carcinoma, using the Ki-67 index as a surrogate. Patients who received a breast cancer diagnosis and had undergone dynamic contrast-enhanced magnetic resonance imaging of the breast for pretreatment evaluation and follow-up were searched retrospectively. We evaluated the MRI studies for perfusion parameters and various categories and compared them to the Ki-67 index. Axillary involvement was categorized as low (N0-N1) or high (N2-N3) according to clinical stage. A total sum of 60 patients' data was included in this study. Perfusion parameters and Ki-67 showed a significant correlation with the transfer constant (Ktrans) (ρ = 0.554 p = 0.00), reverse transfer constant (Kep) (ρ = 0.454 p = 0.00), and initial area under the gadolinium curve (IAUGC) (ρ = 0.619 p = 0.00). The IAUGC was also significantly different between axillary stage groups (Z = 2.478 p = 0.013). Outside of our primary hypothesis, associations between axillary stage and contrast enhancement (x2 = 8.023 p = 0.046) and filling patterns (x2 = 8.751 p = 0.013) were detected. In conclusion, these parameters are potential prognostic markers in patients with moderate Ki-67 indices, such as those in our study group. The relationship between axillary status and perfusion parameters also has the potential to determine patients who would benefit from limited axillary dissection.

Prognostic Significance of Cyclin D1 Expression in Small Intestinal Adenocarcinoma.

Cyclin D1, a critical cyclin-dependent kinase (CDK) 4/6-dependent regulator of G1/S transition, has attracted much interest as a therapeutic target. The cyclin D1 expression in small intestinal adenocarcinomas (SIACs) has not yet been comprehensively studied, owing to the rarity of this tumor. We investigated the clinicopathological and prognostic significance of the cyclin D1 expression in 232 surgically resected primary SIACs through a multi-institutional study. A high expression of cyclin D1 (cyclin D1High) was detected in 145 SIAC cases (63%), which was significantly higher than that in normal small intestinal mucosa (11%). Cyclin D1High was more commonly found in SIACs with a lower T-category and disease stage and KRAS mutation and predicted better patient survival. Multivariate analysis revealed that cyclin D1High, the absence of retroperitoneal seeding and lymphovascular invasion, and the lower N-category were identified as independent prognostic indicators for patients with SIACs. Specifically, cyclin D1High affected patient survival in the lower stage group (stages I and II). In conclusion, cyclin D1 was commonly overexpressed in SIACs, and cyclin D1High acted as a favorable prognostic indicator in patients with SIACs. These findings in SIACs may, thus, be important to further comprehend the mechanism of cyclin D1 in carcinogenesis and to strategize appropriate patient therapies.

Decoding tumor stage by peritumoral and intratumoral radiomics in resectable esophageal squamous cell carcinoma

PurposeTo evaluate the potential application of radiomics in predicting Tumor-Node-Metastasis (TNM) stage in patients with resectable esophageal squamous cell carcinoma (ESCC).MethodsThis retrospective study included 122 consecutive patients (mean age, 57 years; 27 women). Corresponding tumor of interest was identified on axial arterial-phase CT images with manual annotation. Radiomics features were extracted from intra- and peritumoral regions. Features were pruned to train LASSO regression model with 93 patients to construct a radiomics signature, whose performance was validated in a test set of 29 patients. Prognostic value of radiomics-predicted TNM stage was estimated by survival analysis in the entire cohort.ResultsThe radiomics signature incorporating one intratumoral and four peritumoral features was significantly associated with TNM stage. This signature discriminated tumor stage with an area under curve (AUC) of 0.823 in the training set, with similar performance in the test set (AUC 0.813). Recurrence-free survival (RFS) was significantly different between different radiomics-predicted TNM stage groups (Low-risk vs high-risk, log-rank P = 0.004). Univariate and multivariate Cox regression analyses revealed that radiomics-predicted TNM stage was an independent preoperative factor for RFS.ConclusionsThe proposed radiomics signature combing intratumoral and peritumoral features was predictive of TNM stage and associated with prognostication in ESCC.Graphical abstract

Voluntary participation in a terror group and counterterrorism policy

A three-stage game investigates how a government's counterterrorism measures affect potential terrorists interface. In stage 1, the government chooses both proactive and defensive countermeasures, while anticipating the size and attacks of a terrorist group. After radicalized individuals choose whether to join a terrorist group in stage 2, group members then allocate their time between work and terrorism in stage 3. Based on wages and government counterterrorism, the game characterizes the extensive and intensive margins for the terrorist group's size and attacks, respectively. Comparative statics show how changes in wages or population radicalization impact the optimal mix between defensive and proactive countermeasures. Higher (lower) wages favor a larger (smaller) mix of proactive measures over defensive actions. Enhanced radicalization of potential terrorists calls for a greater reliance on defensive actions. Backlash terror attacks stemming from proactive-induced radicalization also affect the mix of counterterrorism actions.

Impact of folate biofortified food supplement on rural women's health and willingness‐to‐pay: A study based on a connected randomized controlled trial—Becker–DeGroot–Marschak experiment in China

AbstractWe examined whether folate‐fortified maize (FFM) improves the health of rural women of childbearing age and whether the health intervention is associated with the consumer willingness‐to‐pay (WTP) for FFM. First, a randomized single‐blind FFM intervention trial was conducted in rural childbearing‐aged women. Participants (n = 55) consumed one stalk of either FFM (treatment group) or ordinary maize (control group) daily. This dietary intervention lasted for 2 months, during which we assessed the participants' serum folate levels at baseline, mid stage (after 1 month), and final stage (after 2 months) to evaluate the health effect of FFM. We found that the serum folate level in the treatment group (13.31 ng/mL) was 3.40 ng/mL higher than that in the control group (9.91 ng/mL) in the final stage of the study. These findings suggest that regular dietary FFM intake significantly increased serum folate levels in rural Chinese women. Second, we further expanded our study by involving 181 local rural women with similar demographic characteristics to participate in a Becker–DeGroot–Marschak (BDM) bidding experiment to measure their WTPs for FFM. Results showed that local consumers were willing to pay 2.82 Chinese Yuan (CNY) per stalk of FFM, approximately 1.21 CNY higher than the price of ordinary maize. And women who participated in and completed the intervention trial had a higher preference for FFM during the evaluation. We provide evidence on the health improvement effect of biofortified foods and shed light on the associated consumer valuation and policy implementation.

99mTc-Labeled FAPI SPECT Imaging in Idiopathic Pulmonary Fibrosis: Preliminary Results.

Idiopathic pulmonary fibrosis (IPF) is associated with a poor prognosis, presenting the most aggressive form of interstitial lung diseases (ILDs). Activated fibroblasts are crucial for pathological processes. Fibroblast activation protein (FAP) inhibitor (FAPI) tracers would be promising imaging agents for these diseases. The purpose of this study was to evaluate a 99mTc-labeled FAPI tracer, 99mTc-HFAPI imaging in IPF patients. Eleven IPF patients (nine males and two females; age range 55-75 year) were included in this pilot study. 99mTc-HFAPI serial whole-body scintigraphy at 5 min, 20 min, 40 min, 1 h, 2 h, 3 h, 4 h, and 6 h was acquired for dynamic biodistribution and dosimetry estimation in seven representative patients. SPECT/CT tomography fusion imaging of the chest region was performed in all patients at 4 h post-injection, which was considered as the optimal acquisition time. Dosimetry was calculated using OLINDA/EXM software (version 2.0; HERMES Medical Solutions). The quantified or semi-quantified standardized uptake values (SUVs) and lesion-to-background ratios (LBRs) of affected lung parenchyma were also calculated. The high-resolution CT (HRCT) stage was determined with visual evaluation, and the total HRCT score of each patient was measured using a weighting factor formula. Pulmonary function tests (PFTs) were recorded as well. Then, the relationships between the 99mTc-HFAPI results, disease extent on HRCT, and PFT results were investigated. Normal physiological uptake of 99mTc-HFAPI was observed mainly in the liver, intestinal tract, pancreas, gallbladder, and to a lesser extent in the spleen, kidneys, and thyroid, with no apparent retention in the blood circulation at the late time point. The mean injected activity of 99mTc-HFAPI was 813.4 MBq (range 695.6-888.0 MBq). No subjective side effects were noticed. The average whole-body effective dose was 0.0041 mSv/MBq per patient. IPF patients exhibited elevated pulmonary 99mTc-HFAPI uptake in abnormal lung regions, which was correlated with fibrotic regions on HRCT. Among different HRCT stage groups, both SUVmax and LBR showed significant differences (p < 0.001). The higher HRCT stage demonstrated significantly higher SUVmax and LBR. A linear correlation between 99mTc-HFAPI uptake and total HRCT score was observed for SUVmax (r = 0.7839, F = 54.41, p = 0.0094) and LBR (r = 0.7402, F = 56.33, p = 0.0092). 99mTc-HFAPI uptake also had moderate correlations with PFT results. Our preliminary data show that the 99mTc-HFAPI SPECT imaging is a promising new imaging modality in IPF patients. Investigations of its clinical value in monitoring disease progression and treatment response are needed in the future.

THU173 Subanalysis By Tanner Stage In Phase 3 Trials In Children And Adolescents With Growth Hormone Deficiency Treated With Lonapegsomatropin

Abstract Disclosure: P. Hofman: Speaker; Self; Novo Nordisk, Eli Lilly &amp; Company. P.S. Thornton: Research Investigator; Self; Ascendis, Pfizer, Inc., Opko, Zealand, Rezolute, Spruce. Other; Self; Johnson &amp;Johnson, Rezolute. U.M. Nadgir: None. P. Saenger: None. E. Chertok: None. E.M. Aghajanova: Speaker; Self; Berlin Hemi Menarini. M. Mao: Employee; Self; Ascendis Pharma. C. Zhao: Employee; Self; Ascendis Pharma. M.R. Ochoa-Maya: Employee; Self; Ascendis Pharma. A. Komirenko: Employee; Self; Ascendis Pharma. A.D. Shu: Employee; Self; Ascendis Pharma. A.K. Maniatis: Advisory Board Member; Self; Novo Nordisk, Ascendis Pharma, Pfizer, Inc. Consulting Fee; Self; Novo Nordisk, Ascendis Pharma. Research Investigator; Self; Novo Nordisk, Pfizer, Inc., OPKO, Ascendis Pharma. Speaker; Self; Novo Nordisk, Ascendis Pharma. Background: Historically, the treatment paradigm in pediatric growth hormone deficiency (GHD) has been to consider increased doses (up to 0.7 mg hGH/kg/week) later in pubertal development to maximize growth outcomes, although this is not a routine recommendation (Mauras et al. JCEM. 2000;85(10):3653-3660, Grimberg et al. Horm Res Paeds. 2016;86(6):361-397). The enliGHten open-label extension trial of once-weekly lonapegsomatropin provided the opportunity to evaluate dosing, efficacy, and safety outcomes in a group of children that includes more advanced Tanner Stages. Lonapegsomatropin (TransCon hGH), a once-weekly prodrug of somatropin approved for the treatment of pediatric GHD by the FDA and EMA, uses TransCon technology to transiently link a parent drug to an inert carrier. In the pivotal phase 3 heiGHt trial, lonapegsomatropin demonstrated noninferior and superior annualized height velocity and a comparable safety profile to daily somatropin in children with GHD. The current analysis explores IGF-1 levels, dose adjustments, and height outcomes by Tanner Stage. Methods: Three phase 3 trials assessed safety and efficacy of lonapegsomatropin: heiGHt (treatment-naïve trial), fliGHt (switch trial), and enliGHten (open-label extension trial). Participants treated in the 3 phase 3 trials were included in this analysis (data snapshot date, September 1, 2021). Data were analyzed by the most recent Tanner Stage at time of assessment in 3 groups: Stage 1 and 2; Stage 3; and Stage 4 and 5. Association between Tanner Stage and IGF-1 SDS measurements, lonapegsomatropin dose, and height SDS were examined using mixed repeated models with Tanner stage as time-varying covariate. Relationship between dose reduction and change in IGF-1 SDS was summarized by Tanner stage. Results: With a mean follow-up of 157 weeks (N=306) on lonapegsomatropin treatment, the analysis showed that later Tanner Stages (3, 4-5) were correlated with higher average IGF-1 SDS (versus Stages 1-2). There was a small difference in lonapegsomatropin dose across Tanner Stage groups, with participants in Stages 4-5 receiving the lowest mean dose (0.19 mg hGH/kg/wk) compared with Stages 1-2 (0.23 mg hGH/kg/wk) or Stage 3 (0.22 mg hGH/kg/wk). Among patients with dose reduction, a decrease of lonapegsomatropin of 0.02 mg hGH/kg corresponded to a consistent drop in average IGF-1 SDS across all Tanner Stages (overall, 0.16 SDS; Stages 1-2, 0.21 SDS; Stage 3, 0.16 SDS; Stages 4-5, 0.21 SDS). Participants across all subgroups experienced continued growth as expected over time. Conclusions: This analysis showed that it was possible to maintain consistent height SDS improvements in children and adolescents in Tanner Stages 3-5 without an increase in the mean lonapegsomatropin dose. Lonapegsomatropin dose reductions resulted in predictable linear IGF-1 decreases across Tanner Stages. Presentation: Thursday, June 15, 2023

Effect of visceral fat area on the accuracy of preoperative CT-N staging of colorectal cancer

ObjectiveTo investigate the effect of visceral fat area (VFA) on the accuracy of preoperative CT-N staging of colorectal cancer. MethodsWe retrospectively reviewed the clinical and imaging data of 385 CRC patients who underwent surgical resection for colorectal cancer between January 2018 and July 2021. Preoperative CT-N staging and imaging features were determined independently by two radiologists. Using postoperative pathology as the gold standard, patients were divided into accurately and incorrectly staged groups, and clinical and imaging characteristics were compared between the two groups. VFA and subcutaneous fat area (SFA) at the L3 vertebral level, sex, age, BMI, tumor location, size, and tumor circumference ratio (TCR) were included. Logistic regression analysis was used to evaluate the independent factors influencing the accuracy of preoperative N staging of colorectal cancer. ResultsOf the 385 patients enrolled, 259 (67.27%) were in the preoperative N-stage accurate staging group, and 126 (32.73%) were in the incorrectly staged group. Univariate analysis showed that there were significant differences in BMI, tumor location, VFA, SFA, size and TCR between the two groups (P<0.05). Logistic regression analysis showed that VFA (95% CI: 1.277, 3.813; P=0.005) and TCR (95% CI: 1.649, 17.545; P=0.005) were independent factors affecting the accuracy of N staging. The optimal cutoff points for VFA and TCR in predicting incorrect staging were 110 cm2 and 0.675, respectively. ConclusionsColorectal cancer patients with lower VFA and higher TCR and preoperative CT-N staging had an increased risk for diagnostic errors.

LncRNA RP11-58O9.2 predicts poor prognosis and promotes progression of non-small cell lung cancer.

Long non-coding RNAs (lncRNAs) play a crucial role in non-small cell lung cancer (NSCLC). This study aimed to investigate the novel biomarker, lncRNA RP11-58O9.2, in patients with NSCLC. RP11-58O9.2 expression in NSCLC cells and tissues was detected by reverse transcription-quantitative polymerase chain reaction. Patient survival was analyzed in relation to RP11-58O9.2 expression levels. RP11-58O9.2 expression was knocked down and endogenous expression was verified in two NSCLC cell lines. Cell proliferation was then assessed by Cell Counting Kit-8 and colony-formation assays, and cell invasion and migration were assessed by Transwell and wound-healing assays, respectively. In vivo experiments were performed in mice, and the combination of RP11-58O9.2 and miR-6749-3p was predicted by miRanda. RP11-58O9.2 was highly expressed in NSCLC cell lines and tissues, and was associated with advanced stage, lymphatic metastasis, and differentiation group. High RP11-58O9.2 levels were also associated with shorter survival. RP11-58O9.2 knockdown inhibited the proliferation, invasion, and migration of lung cancer cells, and tumor growth in mouse xenografts in vivo. RP11-58O9.2 may target and regulate miR-6749-3p. LncRNA RP11-58O9.2 is associated with NSCLC prognosis and promotes lung cancer progression. Further studies are needed to investigate the mechanisms and the regulatory association between RP11-58O9.2 and miR-6749-3p.

A Modified Robust Nomogram for Predicting the Probability of Pelvic Lymph Node Invasion in Localized Prostate Cancer.

Improved predictive models for the risk of pelvic lymph node invasion (LNI) in localized prostate cancer (PCa) is important in decision making of pelvic lymph node dissection (PLND) or whole pelvic RT (WPRT). We aimed to establish a nomogram with improved robustness (Ruijin model) based on pre-treatment information. Continuous localized PCa patients with detailed prostate tumor biopsy information, treated with laparoscopic radical prostatectomy and PLND between 2013 and 2022 in single institution were retrospectively reviewed. A multivariable logistic regression model was fitted and represented the basis for a coefficient-based nomogram of predicting LNI. Comparisons between the Ruijin model and the Roach formula were conducted using the receiver operating characteristic-derived area under the curve (AUC), calibration plot, and decision-curve analyses (DCAs). In total, 624 patients with median age (69.5 year) were included in this analysis. The median number of pelvis lymph nodes removed was 6 (range: 2-30). LNI was found in 35 (5.6%) patients. In the multivariable logistic regression model, total prostate-specific antigen (OR = 1.008, P = 0.025), percentage of cores with the highest-grade PCa (OR = 14.822, P<0.001), clinical stage (III vs. I-II: OR = 7.733, P = 0.008), and biopsy Gleason Grading Group (G3 vs G1-2: OR = 3.152, P = 0.082; G4 vs G1-2: OR = 3.065, P = 0.083; G5 vs G1-2: OR = 5.262, P = 0.008;) were included and formed the basis for the nomogram. The predictive accuracy of Ruijin nomogram in our cohort was 87.7%. Using a cutoff of 4% based on Ruijin nomogram, 395 (63%) PLND would be spared and LNI would be missed in only two (0.5%) patients. The sensitivity, specificity, and negative predictive value associated with the 4% cutoff were 94.3%, 66.7%, and 99.5%, respectively. As compared with the Roach formula, the Ruijin model showed higher AUC (87.7% vs 80.9%, Z = -2.013, P = 0.044), better calibration characteristics, and a higher net benefit at DCA. We developed a novel nomogram for predicting the LNI in localized PCa patients with detailed biopsy information. PLND or WPRT could be avoided in patients with a risk of LNI <4%, so as to spare more than 60% of unnecessary pelvic nodal treatment with a cost of missing only 0.5% LNIs.

Predictive Value of Genomic Classifier Scores and Transcriptomic Data for Prostate Cancer Distant Metastasis Risk: A Multicenter Retrospective Study.

Prostate-specific membrane antigen (PSMA) positron emission tomography (PET) has a greater specificity and sensitivity for detection of extraprostatic prostate cancer than conventional imaging. The Decipher genomic classifier is an established prognostic biomarker being evaluated for its ability to predict systemic treatment intensification. The relationship between Decipher scores and PSMA-based spread remains unknown, as do differences in transcriptomic patterns of PSMA PET-based spread in the de novo vs. recurrent setting. We retrospectively identified patients who (a) had undergone staging with a PSMA PET prior to treatment or for evaluation of recurrence post-radical prostatectomy (RP) at two institutions and (b) had transcriptomic data available from the Genomics Resource for Intelligent Discovery (GRID) database from either biopsy or RP specimens. We classified the PSMA PET pattern of spread using molecular imaging (mi) staging as localized (miT+N0M0), node-positive (miN1M0), distant metastasis (miM1a-c), or negative/non-diagnostic. We used logistic regression to calculate the odds ratios (OR) with 95% confidence intervals (CI) for distant metastasis risk based on Decipher score both pre-treatment and post-RP. As an exploratory analysis, we compared each of the staging groups for differences in important transcriptomic signatures. Kruskal-Wallis and Pearson chi-squared tests were used for continuous and categorical variables, respectively. A total of 315 patients were included in this analysis (n = 164 pre-treatment, n = 151 post-RP). Eighty PSMA PET scans were negative, while 147 were miT+N0M0, 45 were miN1M0, and 43 were miM1a-c. A higher Decipher score was associated with distant metastasis (miM1a-c) on PSMA PET both pre-treatment (OR 1.3 [95% CI: 1.0-1.7] per 0.1 increase in Decipher score, P = 0.05) and post-RP (OR 1.2 [1.0-1.4] per 0.1 increase in Decipher score, P = 0.04). There were higher TP53 mutation (P = 0.01) and cell cycle progression (P = 0.04) signature scores in miM1a-c patients compared to miN1M0 or miT+N0M0 patients. Basal subtype was more prevalent per PAM50 in miM1a-c or miN1M0 patients (36%) than miT+N0M0 patients (19%, P=0.01). Patients with de novo miN1M0 or miM1a disease (n = 19) had higher Decipher scores (0.85 vs 0.57, P = 0.10) and IFNa response (P = 0.08) than patients with recurrent miN1M0 or miM1a disease (n = 35). Higher Decipher scores were associated with distant metastasis on PSMA PET in both the de novo and recurrent setting. Transcriptomic differences in pathways related to proliferation, p53 status, and PAM50 classification were seen when comparing localized, node-positive, and distant metastatic disease. Patients with de novo miN1M0 or miM1a disease may harbor more aggressive disease than those with miN1M0 or miM1a disease at recurrence.

Dawn of Staging for Head and Neck Soft Tissue Sarcoma: Validation of the Novel 8th Edition AJCC T Classification and Proposed Stage Groupings

After decades of stagnation, the 8th edition TNM (TNM8) introduced a new T classification for head and neck (HN) soft tissue sarcomas (STS). New size cutoffs of 2 and 4 cm define T1-3, and a novel T4 category is defined by local invasion of adjoining structures. These size cutoffs had been chosen arbitrarily to advance data collection in this unique disease site since literature showed approximately 70% of HN STS did not reach the previous size threshold (5 cm) for the existing T1 category. The definition of the TNM8 T categories also align with mucosal HN cancers. No stage grouping for HN STS was defined since this new classification required more data collection to derive stage groups. This study aims to validate the TNM8 T classification and to propose stage groupings. Clinical data of all adult (>16 years) HN STS patients treated from 1988 - 2019 with curative intent in our tertiary cancer center were retrieved from a prospective database, and supplemented with chart review. As per TNM8, cutaneous angiosarcoma, embryonal and alveolar rhabdomyosarcoma, Kaposi sarcoma, and dermatofibrosarcoma protuberans were excluded due to their different behavior. Multivariate analysis (MVA) identified prognostic factors for overall survival (OS). Adjusted hazard ratios (AHR) and recursive partitioning analysis (RPA) were used to derive stage groupings. Stage grouping performance for OS was assessed and also compared against the existing TNM8 groups for non-HN STS. A total of 221 patients (N1: 2; M1: 2) were included. Of the 219 M0 patients, 63% were males; median tumor size was 3.0 cm (range: 0.3-14.0); the proportion of TNM8 T1-T4 were 35%, 34%, 26%, and 5%, respectively. Median follow up was 5.9 years. Five-year OS was 79%. MVA confirmed the prognostic value of T category (T4 HR 7.73, 95% CI 3.62-16.5) and grade (G2/3 vs G1 HR 3.7, 95% CI 1.82-7.53), in addition to age (HR 1.03, 95% CI 1.01-1.04) (all p<0.001) for OS. AHR model derived T1-3_Grade 1 as stage 1; T1-3_Grade 2/3 as stage II; and T4_any Grade or any T_N1 as stage III (Table 1); the corresponding 5-year OS was 93%, 73%, and 38%, respectively. Both patients with M1 died within 1.5 years after diagnosis and M1 disease was designated stage IV. The AHR-grouping outperformed the RPA and non-HN TNM8 stage grouping for hazard consistency, hazard discrimination, percent variance explained, hazard difference, and sample size balance. The novel T4 category introduced in TNM8 is associated with a >7 fold increased risk of death. Grade continues to be a critical prognostic factor in HN STS. The TNM8 HN STS T classifications have been validated, and the proposed new stage groupings with TNM8 incorporating grade have excellent performance for OS.

The Effect of Clinical (Anatomical) and Prognostic Stage Groups on Survival in Patients Diagnosed with Breast Cancer: MULTI-Center Study Results, TROD Breast Cancer Study Group

The American Joint Committee on Cancer (AJCC) anatomic stage groups have arguably been the most powerful method in predicting breast cancer outcomes. In the present study, we aimed to determine the differences between anatomical stage and clinical prognostic stage groups, which were obtained by adding biological markers such as histologic tumor grade, estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 status to patients diagnosed with breast cancer, the rate of change in stage, and the effect of stage change on survival. The study protocol was sent to all radiation oncology centers in Türkiye. Four centers enrolled their patients to the study. A total of 1470 stage I-III breast cancer patients with complete information on biological markers (histologic tumor grade, hormone receptor, and c-erb B2 receptor status), haven't been treated with neoadjuvant therapy were included to the study and evaluated retrospectively. The 8th edition of AJCC consolidated biological markers in to clinical prognostic stage groups. Kaplan-Meier curves were used to estimate survival. The log-rank test was used to compare the difference between groups. The Cox proportional-hazards regression model was used to determine the association between anatomic-prognostic stage, disease-free survival (DFS), and overall survival (OS). The median follow-up time was 82 (6-237) months. Median age of the patients was 52 years (20-88) and 865 cases (58.8%) were in the postmenopausal period. The axillary lymph node status was negative (N0) in 765 patients (52.1%). Tumor grade was grade I in 200 patients (13.6%), grade II in 812 patients (55.2%) and grade III in 452 patients (30.7%). Estrogen receptor status was positive in 1247 patients (84.8%), PR status was positive in 1178 patients and Her2-neu status was positive in 207 patients (14.1%). A stage change has been identified in a total of 777 patients (52.9%). Compared with the anatomic stage groups, application of the clinical prognostic stage groups assigned 46.4% cases lower and 6.5% cases higher stage. Five- and ten-year OS and DFS rates of the patients are 73.7%, 44.3% and 91.9%, 86.3% respectively. Age (p<0.001), tumor grade (p<0.001), ER status (p<0.001), PR status (p<0.001), cerbB2 receptor status (p = 0.025) were found to be statistically significant variables in multivariable analysis for OS. For DFS, multivariable analysis showed that age (p = 0.027), tumor grade (p = 0.005), anatomical stage (p<0.001) and assigned to higher stage (p = .001) were statistically significant variables. Hormone receptors and c-erb B2 receptor status are independent variables which impact OS and DFS in our patient group which is mostly consisted of early-stage cases according to anatomical stage. In prognostic staging, upstaging stands out as an independent prognostic factor for DFS. The 8th edition of AJCC prognostic stage groups determines the prognosis much better in our patient cohort.

Prophylactic vs. Reactive Gastrostomy Tube Placement in Patients Treated with Radiotherapy for Head and Neck Cancer

Radiotherapy (RT) for head and neck cancer (HNC) can cause acute toxicities resulting in weight loss warranting alterative enteral access. Some institutions favor prophylactic gastrostomy tube (GT) placement to prevent malnutrition at the forefront, while others choose to reactively place tubes if nutritional deficits arise. Though prophylactic GT placement may limit on-treatment weight loss, this approach may result in unneeded GT placement and may potentially impact long-term swallowing function. Patients with HNC treated with CRT from 2018-2021 were reviewed. GT placement prior to CRT (prophylactic [ppx] tube) was performed at the discretion of the treating radiation oncologist. The remainder of patients were treated with reactive (rx) GT placement in the event of on-treatment weight loss approaching 10-15%. Patient, disease and treatment factors were abstracted from the medical record. Primary endpoints were placement of GT in the rx group, weight change at the end of RT, and time to tube removal. In total, 278 patients were identified; 35 GT-dependent patients and 22 patients with nasal cavity/paranasal sinus cancers were excluded, yielding 221 for analysis. Baseline factors associated with GT group included age, ECOG, baseline swallowing function, primary site, T and N stage, stage group, and treatment year. Treatment factors associated with GT group were: neck target (bilateral v. unilateral/none), concurrent chemotherapy, and total dose. Of the 118 patients in the rx group, 14 (12%) required rx GT placement during or within 30 days of CRT. RT completion rates were similar between groups (95-96%, p = 1). GT was removed at last follow-up in 57% of patients in both groups (p = 1). Percent on-treatment weight loss was -8.9% (SD 6.7) and -7.2% (SD 6.1) for the ppx and rx groups, respectively (p = 0.04). Median Kaplan-Meier estimate of time to GT removal was 7.7 months (95% CI 6.0-11.3) and did not differ between groups (log-rank p = 0.87). Factors associated with the placement of rx GT include: concurrent chemotherapy (yes 20% vs. No 0%, p.0007) and baseline FOIS (5 or less 40% vs. 6 or more 9%, p = 0.02). T stage, N stage, overall stage, postoperative RT, degree of neck irradiation were not associated. Reactive gastrostomy tube placement in patients treated with chemoradiotherapy for head and neck cancer patients is feasible and results in low rates of gastrostomy tube placement. There are no observed differences in on-treatment weight loss between patients treated using a prophylactic versus reactive approach. No differences in time to gastrostomy tube removal were observed.

Cross-Comparison Individual Patient Level Analysis of Three Gene Expression Signatures in Localized Prostate in over 50,000 Men

Risk stratification guides the management of localized prostate cancer. Multiple commercial gene expression biomarkers have been developed to improve estimates of prognosis, however the 22-gene Decipher genomic classifier (22-GC) is the only test with level 1 evidence supporting its use per NCCN guidelines. It is unknown whether other commercial signatures, Oncotype (GPS) or Prolaris (CCP), are sufficiently correlated to negate the differences in evidence supporting these commercial tests. Herein, we aim to perform a cross-comparison of these signatures in a large cohort of patients diagnosed with localized prostate cancer. Patients diagnosed with localized prostate cancer who underwent whole transcriptome gene expression microarray analysis on their primary tumor biopsy specimen were included. The 22-GC score was calculated by Veracyte using a commercially locked model. Individual genes in each of the GPS and CCP gene signatures were identified, and the gene weights in each signature were retrained for prediction of metastasis in a multi-institutional cohort of 1,574 men with long-term outcome data. This was performed to improve correlation performance of GPS and CCP given only the 22-GC was trained for prediction of metastasis. For each of the three signatures, both continuous and categorical scores were calculated. Linear regression and spearman correlations were calculated both on univariable and multivariable analyses adjusting for age, grade group, PSA, and T-stage. A total of 50,881 patients were included (15,379 (30.2%) NCCN low-risk, 14,773 (29.0%) favorable intermediate-risk, 15,544 (30.5%) unfavorable intermediate-risk, and 5,185 (10.2%) high/very high-risk) with a median age of 68 years, and a median PSA of 6.2 ng/mL. On linear regression, the GPS model had poor goodness-of-fit to the 22-GC with an R2 of 0.36, as did the CCP model to the 22-GC with an R2 of 0.32. For CCP, the linear sum of the 31-genes was also tested but had inferior performance (R2 0.28) compared to the reoptimized CCP model. Results were similar on multivariable analysis adjusting for age, PSA, clinical stage and grade group. Spearman correlation between the continuous GPS model scores and the 22-GC was moderate at 0.59, as was the correlation between CCP model and the 22-GC of 0.54. CCP is a measure of proliferation, but in 22-GC high-risk patients, the majority (64.1%) of patients had low-average proliferation and only 35.9% had high proliferation, potentially explaining the lack of strong correlation. There is minimal to moderate correlation between the 22-GC and GPS or CCP gene expression signatures tested. Therefore, these tests should not be viewed as interchangeable, and utilization should be based on the level of evidence supporting each gene expression biomarker.

Longitudinal Assessment of Health-Related Quality of Life in Three Different Hyperphosphatemia Management Groups of Stage III and IV Chronic Kidney Disease Patients.

Health-related quality of life (HRQoL) has emerged as an important outcome measure inpatients with CKD. The lack of prospective studies on HRQoL and its relation with hyperphosphatemia control measures among pre-dialysis patients necessitated the need for this study. This is a prospective, randomized, controlled, open-labelled studythat was conducted for one year on 120 CKD stages III and IV patients divided equally into three groups: Group 1, in which patients under went dietary phosphorus modification; Group 2, in which they were administered calcium-based phosphate binders; and Group 3, in which they were administered non-calcium-based phosphate binders. Patients were assessed for HRQoL, fibroblast growth factor 23 (FGF 23), intact parathyroid hormone (iPTH), phosphorus, and nutritional status, and dietary phosphorus control strategies at 0, 6, and 12 months. HRQoL was measured by using the 36-item Short Form Survey (SF-36) that included a physical component score (PCS) and mental component score (MCS). The scores is ranging from 0 to 100. Higher scores indicate better health status. Usingthe two-wayand one-way repeated measure ANOVA, we analyzed equality of group means, time intervals, and interactions. At baseline, the mean PCS and MCS were equal in three groups. PCS improved significantly (P < 0.00) from baseline to one year in all the three groups: ingroup 1, the PCS score was 66.5 ± 13.5to 75.1 ± 9.76; in group 2, it was 68.9 ± 11.80 to 77.2 ± 7.50; and in group 3, it was 66.2 ± 12.16 to 73.8 ± 9.27. Initially, MCS declined substantially on the sixth month but recovered afterone year. Multiple regression analysis in 13 associated parameters yielded R2 of 13.7% and 18.1% in PCS and MCS, respectively, indicating little contribution of various parameters. There has been a significant positive change in PCSs in three hyperphosphatemia management groups over a period of 12 months. Among the three study groups, the incremental changes in PCS and MCS scores were insignificant.