Spatiotemporal heterogeneity of LMOD1 expression summarizes two modes of cell communication in colorectal cancer.

Cellular communication (CC) influences tumor development by mediating intercellular junctions between cells. However, the role and underlying mechanisms of CC in malignant transformation remain unknown. Here, we investigated the spatiotemporal heterogeneity of CC molecular expression during malignant transformation. It was found that although both tight junctions (TJs) and gap junctions (GJs) were involved in maintaining the tumor microenvironment (TME), they exhibited opposite characteristics. Mechanistically, for epithelial cells (parenchymal component), the expression of TJ molecules consistently decreased during normal-cancer transformation and is a potential oncogenic factor. For fibroblasts (mesenchymal component), the expression of GJs consistently increased during normal-cancer transformation and is a potential oncogenic factor. In addition, the molecular profiles of TJs and GJs were used to stratify colorectal cancer (CRC) patients, where subtypes characterized by high GJ levels and low TJ levels exhibited enhanced mesenchymal signals. Importantly, we propose that leiomodin 1 (LMOD1) is biphasic, with features of both TJs and GJs. LMOD1 not only promotes the activation of cancer-associated fibroblasts (CAFs) but also inhibits the Epithelial-mesenchymal transition (EMT) program in cancer cells. In conclusion, these findings demonstrate the molecular heterogeneity of CC and provide new insights into further understanding of TME heterogeneity.

Artificial intelligence iterative reconstruction in abdominal CT of patients with irregular arm positioning: a case-by-case evaluation.

BACKGROUND: Streak artifacts induced by irregular arm positioning have been an issue in diagnosing the abdomen. PURPOSE: To illustrate the risk of misdiagnosis in abdominal computed tomography (CT) of patients with irregular arm positioning through a case-by-case evaluation and to test if it can be solved by the artificial intelligence iterative reconstruction (AIIR) algorithm. MATERIAL AND METHODS: By reviewing 5220 cases of chest and thoracoabdominal CT, 64 patients with irregular arm positioning were enrolled, whose image data were reconstructed using AIIR in addition to routine hybrid iterative reconstruction (HIR). Lesion detection for livers, spleens, kidneys, gallbladders, and pancreas on AIIR images, performed by two radiologists, was compared with those on HIR images. Discrepancies arising from AIIR images included both cases with additional abnormalities and those with corrections made on previous detections. For cases with discrepancies, artifact scores for organs where discrepancies were found, and contrast-to-noise ratios (CNRs) of cysts with discrepancies were compared between two image sets. RESULTS: Additional abnormalities were detected for 15 cases: additional liver cirrhosis (n=2); additional gallbladder stone (n=1); additional cholecystitis (n=1), additional spleen nodule (n=1); additional kidney cysts (n=8); additional liver cysts (3); and additional spleen cyst (n=1). A spleen contusion was corrected for one case. All involved artifact scores were improved on AIIR images. CNRs of involved liver, kidney, and spleen cysts were improved by up to 539.7%, 538.5%, and 245.5%, respectively. CONCLUSION: Irregular arm positioning may induce a variety of misdiagnoses in abdominal CT, which is almost totally avoidable by the AIIR algorithm.

Association of polycyclic aromatic hydrocarbons with osteoarthritis among adults: an analysis of the NHANES 2001-2016.

OBJECTIVES: Polycyclic aromatic hydrocarbons (PAHs) are environmental endocrine-disrupting compounds, which have been widely recognised as a risk factor for human health. However, the relationship between PAHs exposure and the risk of osteoarthritis has rarely been reported. This study aimed to investigate the association between individual and mixed exposure to PAHs and osteoarthritis. METHODS: In this cross-sectional study, participants aged ≥20 years with data on urinary PAHs and osteoarthritis were extracted from the National Health and Nutrition Examination Survey (NHANES) between 2001 and 2016. Logistic regression analysis was utilised to assess the relationship between individual PAHs exposure and osteoarthritis. The quantile-based g computation (qgcomp) analysis and Bayesian kernel machine regression (BKMR) analysis were performed to assess the effect of mixed exposure to PAHs on osteoarthritis, respectively. RESULTS: A total of 10,613 participants were enrolled, 980 (9.23%) of whom had osteoarthritis. Exposure to high levels of 1-hydroxynaphthalene (1-NAP) [odds ratio (OR)=1.06, 95% confidence interval (CI): 1.01-1.10], 3-hydroxyfluorene (3-FLU) (OR=1.09, 95%CI: 1.02-1.17), and 2-hydroxyfluorene (2-FLU) (OR=1.06, 95%CI: 1.01-1.13) were all associated with greater odds of osteoarthritis after adjusting for age, sex, body mass index, drinking alcohol, and hypertension. The qgcomp analysis showed that the joint weighted value of mixed PAHs exposure (OR=1.11, 95%CI: 1.02-1.22; p=0.017) was significantly related to higher odds of osteoarthritis. The BKMR analysis demonstrated that mixed exposure to PAHs was positively correlated with the risk of osteoarthritis. CONCLUSIONS: Both individual and mixed exposure to PAHs were positively correlated with the risk of osteoarthritis.

IGF2BP3-NRF2 axis regulates ferroptosis in hepatocellular carcinoma.

Poor sensitivity to sorafenib has been an important constraint on the efficacy of targeted therapy in advanced hepatocellular carcinoma (HCC). Therefore, it is particularly important to explore effective therapeutic targets to improve the sensitivity of HCC cells to sorafenib. Upregulation of IGF2BP3 is strongly associated with tumor invasion, early recurrence and poor prognosis in various human cancers, including HCC, but its roles in the sorafenib treatment of HCC remain unclear. In our study, IGF2BP3 knock-down significantly promoted ferroptosis in HCC cells through the evaluation of the Reactive Oxygen Species (ROS), Fe2+ and malondialdehyde (MDA) levels after sorafenib administration. In addition, NRF2 mRNA was identified as an important target of IGF2BP3 by bioinformatics analysis, RNA binding protein immunoprecipitation (RIP) and RNA pulldown experiments. More importantly, IGF2BP3, as an m6A (N6-Methyladenosine) reader, was shown to promote the stability of NRF2 mRNA by reading its m6A modification. Similar results were obtained from in vivo experiments. In summary, our study uncovered the role of IGF2BP3-NRF2 axis on ferroptosis in HCC, providing significant evidence for new anti-cancer strategies aimed at improving the efficacy of sorafenib.

Targeting FAM134B-mediated reticulophagy activates sorafenib-induced ferroptosis in hepatocellular carcinoma.

Ferroptosis is a kind of cell death closely related to selective autophagy, such as ferritinophagy, lipophagy, clockophagy and chaperone-mediated autophagy. However, the role of reticulophagy, which specifically degrades endoplasmic reticulum (ER) fragments (also known as ER-phagy), in ferroptosis regulation is still unclear. In this study, we found that sorafenib (ferroptosis inducer) can effectively activate the receptor protein FAM134B-mediated ER-phagy, and FAM134B knockdown not only blocked ER-phagy but also significantly strengthened cellular sensitivity to ferroptosis without affecting macroautophagy. In vivo experiments also yielded similar results. These evidences provided new clues for ferroptosis regulation. Subsequently, bioinformatic analysis combined with RNA binding protein immunoprecipitation and polyribosome fractionation preliminarily indicated that PABPC1 can interact with FAM134B mRNA and promote its translation. Taken together, this study revealed the role of the PABPC1-FAM134B-ER-phagy pathway on ferroptosis, providing important evidence for novel anti-cancer strategies.

Estrogen alleviates post-traumatic osteoarthritis progression and decreases p-EGFR levels in female mouse cartilage.

OBJECTIVE: To investigate the effect of estrogen on the progression of post-traumatic osteoarthritis (PTOA) in mice and its possible mechanism. METHODS: Twelve-week-old ICR mice were divided into Group A (female control group), group B (ovariectomized(OVX) group), group C (OVX group supplemented with estrogen), and group D (male group) by destabilization of the medial meniscus (DMM)or sham operation. Safranin O staining was performed at 8 weeks and 12 weeks after operation, and the degree of articular cartilage lesion was evaluated using Mankin score. Twelve weeks after the operation, tissue sections were stained to analyze the matrix metalloproteinase 13(MMP13), phosphorylated epidermal growth factor receptor (p-EGFR) expression and apoptosis of chondrocytes. RESULTS: Decreased estrogen can significantly increase the weight of mice in female mice. The degree of cartilage damage in the knee joint on the DMM side of female mice was significantly severer than that on the Sham side. The DMM side also showed higher MMP13 expression and increased apoptotic chondrocytes. The degree of cartilage damage in the knee joint on the DMM side of female mice was significantly reduced after estrogen supplementation, and cartilage damage in the knee joint on the DMM side of female mice was less serious than that of male mice. As estrogen levels decreased, the severity of cartilage erosion in the knee joint on the DMM side was aggravated, and p-EGFR expression in the cartilage surface was also higher in female mice contrast to that in male mice. However, minimal changes in p-EGFR expression in the cartilage surface of bilateral knee joints of male mice were observe. CONCLUSION: Estrogen has a regulatory effect on PTOA and its inhibits the expression of p-EGFR in cartilage on the knee joint surface and has a protective effect on articular cartilage in female mice.

CircBACH1 (hsa_circ_0061395) promotes hepatocellular carcinoma growth by regulating p27 repression via HuR.

In recent years, an increasing number of circular RNAs (circRNAs) have been discovered in hepatocellular carcinoma (HCC). However, the functions of most circRNAs require further investigation. Here, we found that circBACH1 was significantly upregulated in HCC tissues and that high circBACH1 levels were closely associated with poor prognosis. In addition, circBACH1 could promote HCC growth by accelerating cell cycle progression in vitro and in vivo. We next investigated the cellular and molecular mechanisms and discovered that circBACH1 inhibited p27 translation, which influenced cell cycle progression. Moreover, we revealed that circBACH1 could combine directly with HuR using RNA immunoprecipitation assays, pull-down assays, and electrophoretic mobility shift assays. The combination of these molecules facilitated HuR translocation from the nucleus to the cytoplasm according to the fluorescence in situ hybridization and immunofluorescence results. Finally, silencing HuR abrogated circBACH1's inhibition of p27 translation and abolished the circBACH1-induced effect on HCC proliferation. In sum, circBACH1 plays a significant role as an oncogene through the circBACH1/HuR/p27 axis in HCC development.

Comparison of Scoring Systems in Predicting Severity and Prognosis of Hypertriglyceridemia-Induced Acute Pancreatitis.

BACKGROUND: In China, hyperlipidemia is the second major reason of acute pancreatitis. AIMS: Comparison of Scoring Systems in identification patients at risk for severe acute pancreatitis (SAP), pancreatic necrosis (PNec), and infected pancreatic necrosis (IPN) early in the course of hypertriglyceridemia-induced acute pancreatitis (HTG-AP). METHODS: Predictive accuracy of scoring systems was measured by the area under the receiver operating characteristic curve (AUC) in a retrospective study. Pairwise AUC comparisons were performed to calculate the difference between scoring systems. RESULTS: A total of 238 patients diagnosed with HTG-AP were included. Sixty patients (25.2%) were classified as SAP. Twenty-nine patients (12.2%) had evidence of PNec. Nine patients (3.8%) were diagnosed with IPN. One patient (0.4%) died during hospitalization. In predicting SAP in HTG-AP, the AUCs of APACHE-II, SOFA, SIRS, Ranson's, BISAP, and MMS were 0.77, 0.83, 0.73, 0.88, 0.83, and 0.85, respectively; in predicting PNec, were 0.75, 0.77, 0.75, 0.86, 0.80, and 0.75, respectively; and in predicting IPN, were 0.92, 0.86, 0.76, 0.85, 0.84, and 0.87, respectively. Pairwise AUC comparisons revealed that Ranson's, MMS, BISAP, and SOFA had higher accuracy than SIRS, Ranson's and MMS had higher accuracy than APACHE-II in predicting SAP; Ranson's had the same accuracy with BISAP, but higher than other four criteria in predicting PNec; APACHE-II had higher accuracy than SIRS in predicting IPN. CONCLUSIONS: APACHE-II had high performance in predicting IPN, and all other score systems had medium performance in predicting SAP, PNec, and IPN in HTG-AP. Each score has its merit and weakness; BISAP may be the best criterion in predicting severity and prognosis of HTG-AP.

Dynamic changes of lipid profile in severe hypertriglyceridemia-induced acute pancreatitis patients under double filtration plasmapheresis: a retrospective observational study.

BACKGROUND: To investigate the dynamic change of lipid profile under double filtration plasmapheresis (DFPP) in severe hypertriglyceridemia-induced acute pancreatitis (sHTGP) patients and ascertain the association between these changes and the clinical prognosis. METHODS: sHTGP patients admitted within 72 h after disease onset were included, and all the patients received DFPP within 24 h after admission. Lipid profile were detected on admission, consecutive 4 days after DFPP and at discharge. RESULTS: There were 47 sHTGP patients enrolled in this study. At admission, all the parameters of lipid profile changed significantly except for low density lipoprotein. In the first day after DFPP, the serum level of TG, cholesterol and very low density lipoprotein declined significantly, while the high-density lipoprotein (HDL) as well as apoprotein A1 elevated obviously (P < 0.05). TG maintained the downward trend in the following three days and the other parameters kept steady. Linear regression analysis showed that HDL was negatively correlated with the duration of hospitalization among three adjusted models (P = 0.043, P = 0.029, P = 0.025 respectively). CONCLUSION: There was distinct fluctuation of the lipid profile upon the burst of sHTGP and the parameters changed significantly in the first day after DFPP. Among these parameters, HDL may serve as a biomarker for disease prognosis in patients with sHTGP.

The role of double filtration plasmapheresis in hypertriglyceridemic pancreatitis: A propensity score matching analysis.

INTRODUCTION: The role of double filtration plasmapheresis (DFPP) in hypertriglyceridemic pancreatitis (HTGP) is still not well established. Our study aimed to investigate the efficacy of DFPP in reducing triglyceride levels as well as their effects on the outcomes of HTGP. MATERIAL AND METHODS: We retrospectively evaluated the data of patients with HTGP presenting within 72 hours from symptom onset between January 2016 and February 2019. Patients with DFPP treatment were compared with those without DFPP treatment. We used a propensity score matching analysis to reduce confounding factors. RESULTS: Of the 249 patients enrolled, 88 (35.3%) were treated with DFPP. The DFPP was initiated at a median of 7.7 hours from the time of presentation. In the propensity score-matched cohort (n = 100), patients with DFPP had a significantly lower level of triglyceride (P = .034), higher triglyceride reduction (P = .005), and the proportion of triglyceride <5.65 mmol/L (P = .002) by 24 hours after admission when compared with those without DFPP. However, this efficacy did not last until 48 hours after admission. No differences were found in terms of the majority of the clinical outcomes between the two groups, including persistent organ failure (P = .098), local complications (P = .213), hospital stay (P = .657), and in-hospital mortality (P > .999). CONCLUSIONS: HTGP patients with early initiation of DFPP were associated with rapidly and efficiently reduction of triglyceride levels compared with those without DFPP. However, DFPP showed no beneficial effects on improving the clinical outcomes of HTGP.

Elevated triglycerides on admission positively correlate with the severity of hypertriglyceridaemic pancreatitis.

BACKGROUND: Hypertriglycaeridemia has been positively associated with the risk of acute pancreatitis, but whether increased triglyceride (TG) levels are related to the severity of pancreatitis remains unclear. The aim of this study was to assess the relationship between hyperlipidaemia and disease severity of hypertriglycaeridemic pancreatitis. METHODS: From 2016 to 2018, patients with hypertriglyceridaemic pancreatitis presented within 24 hours from symptom onset were retrospectively enrolled. The severity was classified by the Atlanta classification 2012. The demographic and clinical characteristics of patients were compared with respect to severity stratification and different TG categories, respectively. The relationships of admission TG levels and disease severity were assessed with Spearman's rank correlation test and Linear-by-Linear Association test. RESULTS: Among 256 patients included, 125 (48.8%) were diagnosed with mild (MAP), 76 (29.7%) with moderate (MSAP) and 55 (21.5%) with severe acute pancreatitis (SAP). The mean TGs (standard derivation) on admission in patients with MAP, MSAP and SAP were 21.6 (15.2) mmol/L (1913 [1346] mg/dL), 34.6 (22.6) mmol/L (3065 [2002] mg/dL) and 41.5 (32.5) mmol/L (3676 [2879] mg/dL), respectively (P < .001). Patients were then categorised based on their TG levels. TG categories had a strong positive correlation with the disease severity (Rho = 0.34, P < .001). Positive trend for the association across increasing TG categories and SAP was observed (P < .001). CONCLUSIONS: Elevated serum TG levels at the time of admission seem to correlate positively with the severity of hypertriglyceridaemia-induced acute pancreatitis.

Differentiating Transition Zone Cancers From Benign Prostatic Hyperplasia by Histogram Analysis of Apparent Diffusion Coefficient Maps With Standard and Ultrahigh b-value Diffusion-weighted MR Imaging.

OBJECTIVE: To compare the diagnostic performance of standard and ultrahigh b-value Diffusion-weighted Imaging (DWI) using volumetric histogram analysis in differentiating transition zone (TZ) cancer from benign prostatic hyperplasia (BPH). METHODS: 57 TZ cancer and 61 BPH patients received standard (1000 s/mm) and ultrahigh b-value (2000 s/mm) DWI. The diagnostic ability of ADC histogram parameters derived from two DWI for differentiating TZ cancer from BPH was determined by receiver operating characteristic curve. RESULTS: Median, minimum, the 10th, 25th percentile ADC in both ADC1000 and ADC2000 and skewness in ADC2000 had significant differences between TZ cancer and BPH (for all, P < 0.05).The 10th percentile ADC showed highest area under the ROC curve (AUC) in both ADC1000 and ADC2000.The 10th percentile ADC of ADC2000 showed significantly higher AUC than did ADC1000 (P = 0.0385). CONCLUSIONS: The 10th percentile ADC obtained from ultrahigh b-value DWI performed better for differentiating TZ cancer from BPH.

Alterations in diaphragmatic function assessed by ultrasonography in mechanically ventilated patients with sepsis.

PURPOSE: To assess alteration of diaphragmatic function by ultrasonography in a population of mechanically ventilated patients with or without sepsis. METHODS: We performed a prospective, 6-month, single-center, observational cohort study. Mechanically ventilated septic and nonseptic patients were studied within 24 hours following intubation and before the moment of ventilator liberation. Diaphragm thickness and contractile activity (quantified by diaphragmatic thickening fraction, DTF) were measured by ultrasonography at the zone of apposition. Intraobserver and interobserver reproducibility were measured. RESULTS: Fifty-two critically ill patients were included, 28 with sepsis and 24 without sepsis. Upon initiation of ventilation, DTF was lower in septic than that in nonseptic patients (P = 0.03). No difference was observed between septic and nonseptic patients for diaphragm thickness. Mean 188 ± 111 hours after the first measurement, both diaphragm thickness and DTF decreased significantly compared with first measurements in septic and nonseptic patients, all P < 0.001. Diaphragm thickness decreased by 9.1 ± 10.7% in nonseptic and by 16.0 ± 13.5% in septic patients, P = 0.049. DTF decreased by 15.2 ± 21.3% in nonseptic and by 30.7 ± 22.0% in septic patients, P = 0.013. CONCLUSIONS: Mechanically ventilated patients with sepsis were associated with an earlier and more severe diaphragm dysfunction compared with patients without sepsis.

Prediction of Clinical Pathologic Prognostic Factors for Rectal Adenocarcinoma: Volumetric Texture Analysis Based on Apparent Diffusion Coefficient Maps.

Texture analysis has been used to characterize and measure tissue heterogeneity in medical images. The purpose of this study was to investigate the potential of texture features derived from apparent diffusion coefficient (ADC) maps, to serve as imaging markers for predicting important histopathologic prognostic factors in rectal cancer. One hundred patients of rectal cancer received 3 T preoperative magnetic resonance imaging including diffusion-weighted imaging (DWI). Skewness, kurtosis, uniformity from the histogram and entropy, energy, inertia, correlation from gray-level co-occurrence matrix (GLCM) derived from whole-lesion volumes were measured. Independent sample t-test or Mann-Whitney U-test and receiver operating characteristic (ROC) curves were used for statistical analysis. Uniformity, energy and entropy were significantly different (p = 0.026, 0.001, and 0.006, respectively) between stage pT1-2 and pT3-4 tumors. Skewness, kurtosis and correlation were significantly different (p = 0.000, 0.006, and 0.041, respectively) between grade 1-2 and grade 3 tumors. Energy and entropy (p = 0.008 and 0.033, respectively) could significantly differentiate negative circumferential resection margin (CRM) from positive CRM. Furthermore, predicted probabilities derived by logistic regression analysis yielded greater area under the curve (AUC) in differentiating pT3-4 stage and grade 3 grade tumors. Texture features derived from ADC maps may useful to predict important histopathologic prognostic factors of rectal cancer.

Study on mechanism about long noncoding RNA MALAT1 affecting pancreatic cancer by regulating Hippo-YAP signaling.

By investigating the migration and invasion ability in pancreatic cancer, this study probed into the lncRNA MALAT1 molecular mechanism on Hippo-YAP signaling. The expression of lncRNA MALAT1 in PC tissues and cells was detected by qRT-PCR and Western blot. The effect of si-MALAT1 on proliferation was determined by CCK-8 assay. Cell apoptosis, migration, and invasion were respectively detected by flow cytometry assay, wound healing assay, and transwell assay. Western blot and immunohistochemistry were successively used for detecting LATS1 and YAP1 expression in pancreatic cancer tissues. The microarray analysis determined that lncRNA MALAT1 in pancreatic cancer was highly expressed. LncRNA MALAT1 presented an extremely high expression level in pancreatic cancer tissues and cells. After transfected with si-MALAT1, the proliferation of AsPC-1 cells decreased, induce apoptosis of AsPC-1 cells, and migration and invasion ability were reduced. The tendency of LATS1 expression level was down-regulated and YAP1 show the opposite trend in the Hippo-YAP signaling. The in vivo assay was found that the tumor to be small in size and volume, and the expression of Ki-67 was decreased. High expression of lncRNA MALAT1 in PC disorder the proliferation, apoptosis, and migration and invasion ability via influence Hippo-YAP signaling pathway.

Low-intensity extracorporeal shock wave therapy promotes myogenesis through PERK/ATF4 pathway.

AIM: Stress urinary incontinence (SUI) is a significant health problem for women. Treatments employing muscle derived stem cells (MDSCs) may be a promising approach to this prevalent, bothersome condition, but these treatments are invasive and require collection of cells from one site for injection into another. It is also unknown whether or not these cells establish themselves and function as muscle cells in the target tissues. Alternatively, low-intensity extracorporeal shock wave therapy (Li-ESWT) is non-invasive and has shown positive outcomes in the treatment of multiple musculoskeletal disorders, but the biological effects responsible for clinical success are not yet well understood. The aim of this study is to explore the possibility of employing Li-ESWT for activation of MDSCs in situ and to further elucidate the underlying biological effects and mechanisms of action in urethral muscle. METHODS: Urethral muscle derived stem cells (uMDSCs) were harvest from Zucker Lean (ZUC-LEAN) (ZUC-Leprfa 186) rats and characterized with flow cytometry. Li-ESWT (0.02 mJ/mm2 , 3 Hz, 200 pulses) and GSK2656157, an inhibitor of PERK pathway, were applied to L6 rat myoblast cells. To assess for myotube formation, we used immunofluorescence staining and western blot analysis in uMDSCs and L6 cells. RESULTS: The results indicate that uMDSCs could form myotubes. Myotube formation was significantly increased by the Li-ESWT as was the expression of muscle heavy chain (MHC) and myogenic factor 5 (Myf5) in L6 cells in vitro. Li-ESWT activated protein kinase RNA-like ER kinase (PERK) pathway by increasing the phosphorylation levels of PERK and eukaryotic initiation factor 2a (eIF2α) and by increasing activating transcription factor 4 (ATF4). In addition, GSK2656157, an inhibitor of PERK, effectively inhibited the myotube formation in L6 rat myoblast cells. Furthermore, GSK2656157 also attenuated myotube formation induced by Li-ESWT. CONCLUSION: In conclusion, this experiment reveals that rat uMDSCs can be isolated successfully and can form myotubes in vitro. PERK/ATF4 pathway was involved in myotube formation, and L6 rat myoblast cells were activated by Li-ESWT to form myotubes. These findings suggest that PERK/ATF4 pathway is activated by Li-ESWT. This study elucidates one of the biochemical pathways responsible for the clinical improvements seen after Li-ESWT. It is possible that this information will help to establish Li-ESWT as an acceptable treatment modality and may help to further refine the use of Li-ESWT in the clinical practice of medicine.

In Situ Activation of Penile Progenitor Cells With Low-Intensity Extracorporeal Shockwave Therapy.

BACKGROUND: We previously reported that progenitor cells, or stem cells, exist within penile tissue. We hypothesized that acoustic wave stimulation by low-intensity extracorporeal shockwave therapy (Li-ESWT) would activate local stem or progenitor cells within the penis, producing regenerative effects. AIMS: To study the feasibility of in situ penile progenitor cell activation by Li-ESWT. METHODS: We performed a cohort analysis of young and middle-age male Sprague-Dawley rats treated with 5-ethynyl-2'-deoxyuridine (EdU) pulse followed by Li-ESWT. In addition, Li-ESWT was applied to cultured Schwann cells and endothelial cells to study the molecular mechanism involved in cell proliferation. Thirty minutes before Li-ESWT, each rat received an intraperitoneal injection of EdU. Li-ESWT was applied to the penis at very low (0.02 mJ/mm2 at 3 Hz for 300 pulses) or low (0.057 mJ/mm2 at 3 Hz for 500 pulses) energy levels. The endothelial and Schwann cells were treated with very low energy (0.02 mJ/mm2 at 3 Hz for 300 pulses) in vitro. OUTCOMES: At 48 hours or 1 week after Li-ESWT, penile tissues were harvested for histologic study to assess EdU+ and Ki-67+ cells, and cell proliferation, Ki-67 expression, Erk1/2 phosphorylation, translocation, and angiogenesis were examined in cultured Schwann and endothelial cells after Li-ESWT. RESULTS: Li-ESWT significantly increased EdU+ cells within penile erectile tissues (P < .01) at 48 hours and 1 week. There were more cells activated in young animals than in middle-age animals, and the effect depended on dosage. Most activated cells were localized within subtunical spaces. In vitro studies indicated that Li-ESWT stimulated cell proliferation through increased phosphorylation of Erk1/2. CLINICAL TRANSLATION: The present results provide a possible explanation for the clinical benefits seen with Li-ESWT. STRENGTHS AND LIMITATIONS: The main limitation of the present project was the short period of study and the animal model used. Li-ESWT could be less effective in improving erectile function in old animals because of the decreased number and quality of penile stem or progenitor cells associated with aging. CONCLUSION: Li-ESWT activation of local penile progenitor cells might be one of the mechanisms that contribute to the beneficial effects of shockwave treatment for erectile dysfunction, which represents a non-invasive alternative to exogenous stem cell therapy. Lin G, Reed-Maldonado AB, Wang B, et al. In Situ Activation of Penile Progenitor Cells With Low-Intensity Extracorporeal Shockwave Therapy. J Sex Med 2017;14:493-501.

Impaired contractility of the circular striated urethral sphincter muscle may contribute to stress urinary incontinence in female zucker fatty rats.

AIM: Obesity has been an independent risk factor for female stress urinary incontinence (SUI), the mechanism of this association remains unknown. The aim of this study is to validate the hypothesis that urethral dysfunction is a possible contributor to SUI in obese women. METHODS: Ten Zucker Fatty (ZF) (ZUC-Leprfa 185) and 10 Zucker Lean (ZL) (ZUC-Leprfa 186) female rats at 12-week-old were used in this experiment. The urethral sphincter rings were harvested from the bladder neck through to the most proximal 2/3 regions. In the organ bath study, single pulses of electrical field stimulation (EFS) were applied. For the fatiguing stimulation, repeated multi-pulse EFS with 70 mA were applied at frequency of 5 Hz for 5 min. Caffeine-containing Krebs' solution was administrated to contract the urethra until the contraction began to reach a plateau for 10 min. We performed immunofluorescence staining of the urethra after the experiment was finished. RESULTS: Compared to ZL controls, ZF rats had significantly impaired muscle contractile activity (MCA) (P < 0.05). Also, ZF rats presented early fatiguing of MCA and had a significantly greater percentage of MCA decline from baseline in the fatiguing test (37.7% vs 25.6%, P < 0.05). The plateau of maximal MCA induced by caffeine in ZF rats was significantly lower than ZL controls (0.22 vs 0.36, P < 0.05). CONCLUSIONS: This novel study showed that obese female rats had significantly impaired contractile properties of striated urethral sphincter, suggesting urethral dysfunction could be an important contributor to SUI in obesity.

Anatomical principles for minimally invasive reconstruction of the acromioclavicular joint with anchors.

PURPOSE: The aim of this study was to evaluate the outcome of a minimally invasive surgical technique for the treatment of patients with acromioclavicular joint dislocation. METHODS: Sixteen patients with complete acromioclavicular joint dislocation were enrolled in this study. All patients were asked to follow the less active rehabilitation protocol post-operatively. Computed tomography with 3-D reconstruction of the injured shoulder was performed on each patient post operatively for the assessment of the accuracy of the suture anchor placement in the coracoid process and the reduction of the acromioclavicular joint. Radiographs of Zanca view and axillary view of both shoulders were taken for evaluating the maintenance of the acromioclavicular joint reduction at each follow-up visit. The Constant shoulder score was used for function assessment at the final follow-up. RESULTS: Twenty seven of the 32 anchors implanted in the coracoid process met the criteria of good position. One patient developed complete loss of reduction and another had partial loss of reduction in the anteroposterior plane. For the other 14 patients, the mean Constant score was 90 (range, 82-95). For the patients with partial and complete loss of reduction, the Constant score were 92 and 76 respectively. All of them got nearly normal range of motion of the shoulders and restored to pre-operative life and works. CONCLUSION: With this minimally invasive approach and limited exposure of the coracoid, a surgeon can place the suture anchors at the anatomical insertions of the coracoclavicular ligament and allow the dislocated joint reduced and maintained well. LEVEL OF EVIDENCE: Level IV, Case series; therapeutic study.

The effects of STAT3 and Survivin silencing on the growth of human bladder carcinoma cells.

Despite accumulating evidence suggesting a critical role of signal transducer and activator of transcription 3 (STAT3) and Survivin in human bladder cancer, the effects of silencing these genes on the proliferation of T24 bladder carcinoma cells remain unknown. Here, we investigated the inhibitory effects of STAT3 or Survivin silencing on the in vitro and in vivo growth of human T24 bladder carcinoma cells. Small interfering RNA (siRNA) vectors targeting STAT3, Survivin, or both genes were designed and synthesized. The recombinant plasmid DNA constructs were confirmed by DNA sequencing. They were then transiently transfected into T24 cells, and the mRNA and protein expressions of STAT3 and Survivin were determined using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot, respectively. Cell proliferation was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The in vivo growth-inhibition effects of the siRNA vectors were assessed using a bladder cancer mouse model. The tumor weight and size was recorded 4 weeks post-inoculation. We successfully synthesized four siRNA vectors targeting STAT3 and four vectors targeting Survivin. The STAT3-3 and Survivin-4 siRNA constructs were most efficient in reducing STAT3 or Survivin expression, respectively. We then constructed a vector containing these two sequences together (STAT3-Survivin siRNA) and found that the single vector efficiently silenced STAT3 and Survivin expression. Moreover, silencing of STAT3 or Survivin significantly suppressed the in vitro and in vivo proliferation of T24 cells compared to the controls (P<0.05). Our findings indicated that the downregulation of STAT3 or Survivin can suppress the proliferation of T24 bladder cancer cells. Moreover, no additive effects were observed when STAT3 and Survivin were knocked down together, suggesting that they work in the same signaling pathway in T24 cells. These results provide valuable insights into understanding the pathways involved during the tumorigenesis of bladder cancer.