Tumor immune microenvironment-based clusters in predicting prognosis and guiding immunotherapy in breast cancer.

The development and progression of breast cancer (BC) depend heavily on the tumor microenvironment (TME), especially tumor infiltration leukocytes (TILs). TME-based classifications in BC remain largely unknown and need to be clarified. Using the bioinformatic analysis, we attempted to construct a prognostic nomogram based on clinical features and TME-related differentially expressed genes (DEGs). We also tried to investigate the association between the prognostic nomogram and clinical characteristics, TILs, possible signaling pathways, and response to immunotherapy in BC patients. DEGs for BC patients were identified from The Cancer Genome Atlas Breast Invasive Carcinoma database. TME-related genes were downloaded from the Immunology Database and Analysis Portal. After intersecting DEGs and TME-related genes, 3985 overlapping TME-related DEGs were selected for non-negative matrix factorization clustering, microenvironment cell populations-counter (MCP-counter), LASSO Cox regression, tumor immune dysfunction, and exclusion (TIDE) algorithm analyses. BC patients were divided into three clusters based on the TME-related DEGs and survival data, in which cluster 3 had the best overall survival (OS). Of note, cluster 3 exhibited the highest infiltration or lowest infiltration of CD3+ T-cells, CD8+ T-cells, cytotoxic lymphocytes, B-lymphocytes, monocytic lineage, and myeloid dendritic cells (MDCs). A total of 33 TME-related DEGs were identified as a prognostic gene signature by the LASSO regression analysis. The prognostic gene signature separated BC patients into low- and high-risk groups with significant differences in OS (p<0.01) and demonstrated powerful effectiveness (TCGA all group: 1-year area under the curve [AUC] = 0.773, 3-year AUC = 0.770, 5-year AUC = 0.792). By integrating demographic features, tumor-node metastasis (TNM) stages, and prognostic gene signature, we constructed a nomogram with better predictive value than other clinical features alone. TMErelated DEGs in the low-risk BC patients (with better OS) were enriched in chemokine, cytokine-cytokine receptor interaction, and JAK-STAT and Toll-like receptor signaling pathways. BC patients in the low-risk group exhibited higher TIDE scores associated with worse immune checkpoint blockade response. A prognostic nomogram based on TME-related DEGs and clinical characteristics could predict prognosis and guide immunotherapy in BC patients.

PI3K/Akt and Wnt/ß-catenin Signaling Cross-regulate NF-κB Signaling in TNF-α-induced Human Lgr5+ Intestinal Stem Cells.

BACKGROUND/AIM: Intestinal stem cells (ISCs) are responsible for intestinal proliferation, differentiation, and neoplasia, and also play a crucial role in inflammation. Thus, it is important to investigate the effect of TNF-α on the activities of NF-κB, PI3K/Akt, and Wnt/ß-catenin signaling pathways. MATERIALS AND METHODS: The Lgr5+ intestinal cells were isolated using fluorescence-activated cell sorting from NCM460 spheroid cells, and the potential molecular mechanisms were investigated via short hairpin RNA (shRNA) transfection or the use of an inhibitor. RESULTS: The Lgr5+ cells were termed ISCs because of the higher expression of stem cell genes, including Sox2, Nanog, Oct4, Lgr5, and CD133. The Lgr5+ ISCs had a higher proliferation capacity, invasive ability, and drug resistance to 5-fluorouracil, as well as higher expression levels of anti-apoptotic proteins but lower expression levels of pro-apoptotic proteins, compared with Lgr5~ cells. The PI3K/Akt and Wnt/ß-catenin pathways were triggered by the TNF-α-induced activation of NF-κB signaling. Notably, when p65 expression was knocked-down via shRNA transfection in Lgr5+ ISCs, the TNF-α-induced activation of the NF-κB, PI3K/Akt, and Wnt/ß-catenin pathways were reversed. The same effect was observed with regards to ß-catenin shRNA transfection. Moreover, the Akt inhibitor MK2206 inhibited the TNF-α-induced activation of the PI3K/Akt pathway, as well as the NF-κB and Wnt/ß-catenin pathways. CONCLUSION: PI3K/Akt and Wnt/ß-catenin signaling cross-regulate NF-κB signaling in TNF-α-induced human Lgr5+ ISCs.

Mortality Effects of Hypothetical Interventions on Physical Activity and TV Viewing.

INTRODUCTION: Long-term effects of physical activity and television (TV) viewing on mortality have been inferred from observational studies. The associations observed do not allow for inferences about the effects of population interventions and could be subject to bias due to time-varying confounding. METHODS: Using data from the Australian Diabetes, Obesity and Lifestyle Study, collected in 1999-2000 (T0), 2004-2005 (T1), and 2011-2012 (T2), we applied the parametric g-formula to estimate cumulative risks of death under hypothetical interventions on physical activity and/or TV viewing determined from self-report while adjusting for time-varying confounding. RESULTS: In the 6377 participants followed up for 13 yr from 2004 to 2005 to death or censoring in 2017, 781 participants died. The observed cumulative risk of death was 12.2%. The most effective hypothetical intervention was to increase weekly physical activity to >300 min (risk ratio (RR), 0.66 (0.46-0.86) compared with a "worst-case" scenario; RR, 0.83 (0.73-0.94) compared with no intervention). Reducing daily TV viewing to <2 h in addition to physical activity interventions did not show added survival benefits. Reducing TV viewing alone was least effective in reducing mortality (RR, 0.85 (0.60-1.10) compared with the worst-case scenario; RR, 1.06 (0.93-1.20) compared with no intervention). CONCLUSIONS: Our findings suggested that sustained interventions to increase physical activity could lower all-cause mortality over a 13-yr period, and there might be limited gain from intervening to reduce TV viewing time in a relatively healthy population.

Heavy Metals in Grains from Jilin Province, China, and Human Health Risk.

ABSTRACT: Heavy metals are an indispensable part of industrial and agricultural development. As the cradle of China's industry and an important province for agricultural production, Jilin Province has been an area of concern about heavy metal pollution in the local environment and grains. In this study, we focused on four heavy metals that are harmful to humans: arsenic (As), cadmium (Cd), methylmercury (MeHg), and inorganic arsenic (iAs). We determined the contents of these metals in 341 grain samples by using graphite furnace atomic absorption spectrometry and liquid chromatography-atomic fluorescence spectrometry and compared our results with the limit value of national standards. To evaluate the potential risk to human health, we determined the target hazard quotient and hazard index. Heavy metals were detected at these rates, from high to low: Cd (48%) > iAs (20.8%) > MeHg (4.6%) > Pb (3%). Most of these values are far below the limit of national standards. The target hazard quotient and hazard index were both smaller than 1; thus, we conclude that heavy metal pollution in grains in Jilin Province is not serious and that people are not at high risk from heavy metals in grains.

Moderate Exercise Enhances Endothelial Progenitor Cell Exosomes Release and Function.

PURPOSE: Exercise has cardiovascular benefits which might be related to endothelial progenitor cells (EPC). Meanwhile, there is evidence suggesting that EPC-derived exosomes (EPC-EX) promote vascular repair and angiogenesis through their carried microRNA (miR)-126. In this study, we investigated whether exercise could increase the levels of circulating EPC-EX and their miR-126 cargo, and by which promote the protective function of EPC-EX on endothelial cells (EC). METHODS: Plasma EPC-EX from sedentary, low, or moderate exercise mice, respectively, denoted as EPC-EX, EPC-EX, and EPC-EX, were isolated using microbead-based sorting techniques and characterized by nanoparticle tracking analysis, Western blot, and quantitative real-time polymerase chain reaction assessments of biomarkers and miR-126. High glucose (25 mM) with hypoxia (1% O2) was used for inducing an EC injury model. The injured EC were treated by coculturing with vehicle, EPC-EX, EPC-EX, EPC-EX, or EPC-EX + anti-miR-126. After that, EC were used for flow cytometry analysis of apoptosis, assessments of tube formation and migration, and measurements of miR-126 level and its downstream sprouty-related protein-1 (SPRED1) and vascular endothelial growth factor (VEGF). RESULTS: 1) Isolated EPC-EX positively expressed exosomal markers (CD63 and Tsg101) and EPC markers (CD34 and VEGFR2). 2) Exercise intensity dependently elevated plasma level of EPC, EPC-EX/EPC ratio, and miR-126 expression in EPC and EPC-EX. 3) Injured EC displayed apoptosis increment, angiogenic dysfunction and miR-126 reduction. 4) EPC-EX had better effects than EPC-EX and EPC-EX on alleviating those changes of injured EC, accompanied with SPRED1 downregulation and VEGF upregulation. 5) The effects of EPC-EX were abolished by miR-126 knockdown. CONCLUSIONS: Our data demonstrate that exercise can increase EPC-EX release and miR-126 level and enhance the effects of EPC-EX on protecting EC against injury through the SPRED1/VEGF pathway.

What is the Optimal Age for Students to Receive Cardiopulmonary Resuscitation Training?

OBJECTIVES: Training students has been proven to be the optimal way to deliver cardiopulmonary resuscitation (CPR) skills. However, it is somehow unknown whether or not the current recommendations appropriate for Caucasian students are also suitable for East Asian students. The purpose of this study is to explore the best age for East Asian students to receive CPR training. METHODS: Students were recruited from six schools. Students participated in a standard CPR training program provided by tutors. Each student attended a 60-minute training session with a manikin. After being trained, within one hour, the student's compression quality was assessed. RESULTS: A total of 360 students who constituted 12 continuous grades were recruited for this study. Adequate compression depth and satisfactory compression rate with correct hand position could be achieved since the age of 12. However, successful compression rate and complete release could be achieved since the younger age of six. CONCLUSIONS: Current recommendations for Caucasian students to cultivate a full-capacity CPR rescuer at the age of 12 are also appropriate for East Asian students. However, the optimal age for students to receive CPR training should be decided based on evidence and importance assessment of CPR.He D, Huang K, Yang Y, Jiang W, Yang N, Yang H. What is the optimal age for students to receive cardiopulmonary resuscitation training? Prehosp Disaster Med. 2018;33(4):394-398.

Effect of synchronous solitary bone metastasectomy and lung cancer resection on non-small cell lung cancer patients.

Lung cancer is the most frequent cause of cancer-associated mortality among men and women globally. The skeleton is one of the most common metastatic sites. The majority of patients exhibiting bone metastases are treated using systemic therapy or symptom-based palliative approaches without surgery. The present study attempted to improve the therapeutic effects of synchronous surgeries in resectable non-small cell lung cancer patients exhibiting solitary bone metastasis. A total of 5 patients underwent synchronous lung cancer resections and solitary bone metastasectomies between October 2009 and November 2011 in the Department of Cardiothoracic Surgery, Shanghai Sixth People's Hospital (Shanghai, China). All patients had received fluorodeoxyglucose positron emission tomography-computed tomography or bone scintigraphy to demonstrate the presence of solitary bone metastasis and to exclude the presence of metastases at alternative sites. The patients received standard lung cancer and mediastinal lymph node resections. In addition, bone lesions were assessed by orthopedists and operated on synchronously with standard procedures. Following surgery, all patients were administered standard chemotherapeutic regimens. Perioperative indicators, including time for thoracic drainage, length of hospital stay, incidence of post-operative complications and progression-free survival (PFS) time, were observed. The average time for post-operative drainage was 4.6±1.1 days, and the average length of post-operative hospitalization was 8.8±2.2 days. All procedures were performed safely with no serious complications. The PFS of the patients was 13.2±7.7 months. While 2 patients presenting with spinal metastases succumbed at ~1 year post-surgery, the remaining 3 patients presenting with limb bone metastases survived for >16 months post-surgery, and were alive at the last follow-up. In conclusion, the present study indicated that a synchronous metastasectomy and lung tumor resection is a safe method of treatment. The PFS time and survival results demonstrated that on the rare occasion that a patient exhibits solitary bone metastasis, aggressive surgical treatment may be a potential therapeutic option.

Hepatoid adenocarcinoma of the stomach: A case report and review of the literature.

Hepatoid adenocarcinoma is a rare tumor, associated with an extremely poor prognosis. In the present report, a 72-year-old female presented to Huashan Hospital (Shanghai, China) with upper abdominal discomfort and, following endoscopy, an ulcerated lesion was observed in the stomach. A computed tomography scan revealed a mass in the antrum of the stomach and the α-fetoprotein level was normal. Sections of the mass exhibited malignant cells arranged in a solid to trabecular pattern; the cells were polygonal shaped with well-defined cytoplasmic borders. Immunohistochemical analysis was performed and the cells were positive for Hep1, CK, CK8, CK18 and P53. The histological features together with the immunohistochemical findings were diagnostic of a hepatoid adenocarcinoma of the stomach. Immunohistochemical studies may aid in the identification of the characteristic features and prevent the misdiagnosis of this tumor.

Nedaplatin- versus cisplatin-based chemotherapy in the survival time of patients with non-small cell lung cancer.

Nedaplatin (NDP) has been extensively used to treat patients with non-small cell lung cancer (NSCLC) in the last decade. The present study compared the survival benefits of NDP and cisplatin (DDP) in the treatment of NSCLC. Patients (n=392) with NSCLC were treated with at least two cycles of platinum-based chemotherapy. Among these patients, 202 received DDP-based chemotherapy, and 190 received NDP-based chemotherapy. The overall survival time of the two groups and the toxicity of drugs were analyzed. The results showed that only the chemotherapy cycle duration was found to be statistically different between DDP and NDP groups in all the characteristics. The mean chemotherapy duration was 3.3 cycles in the DDP group, and 4.1 cycles in the NDP group (χ2=20.206, P<0.001). Additionally, the chemotherapy cycle number was also an independent predictive factor for the overall survival time in the multivariate analysis (HR=0.539, P<0.001). The median survival time (MST) was 15 months in the DDP group, and 20 months in the NDP group (χ2=5.189, P=0.023). The 1-, 2- and 3-year overall survival rates were 62.4, 25.7 and 15.8%, and 78.9, 38.9, and 16.8% in the DPP and NDP groups, respectively. The incidence of grade 3-4 nausea/vomiting, anorexia and weight loss was higher in the DDP compared to the NDP group (36.1 vs. 8.4%, 17.3 vs. 5.8%, and 9.9 vs. 1%, respectively). In conclusion, NDP-based chemotherapy had a survival benefit compared to DDP-based chemotherapy for NSCLC patients, due to the lower toxicity of NDP, which renders this drug more tolerable, thus allowing patients to undergo more cycles of chemotherapy.

JAK/STAT3 signaling pathway mediates endothelial-like differentiation of immature dendritic cells.

Endothelial-like differentiation (ELD) of dendritic cells (DCs) is a poorly understood phenomenon. The present study evaluated the effect on the ELD of DCs by using human esophageal squamous cell carcinoma (ESCC) cells with high or poor differentiation. The results demonstrated that KYSE450 (highly differentiated) and KYSE70 (poorly differentiated) cell supernatants induce the differentiation of immature DCs (iDCs), derived from healthy adult volunteers, away from the DC pathway and towards an endothelial cell (EC) fate. This effect was strongest in the cells treated with the KYSE70 supernatant. During the ELD of iDCs, sustained activation of JAK (janus tyrosine kinase)/STAT3 (signal transducer and activator of transcription 3) signaling was detected. Incubation of iDCs with the JAK inhibitor, AG490 blocked JAK/STAT3 phosphorylation and iDC differentiation. These results suggested that the JAK/STAT3 signaling pathway mediates ELD of iDCs. Furthermore, the poorly differentiated ESCC cells may have a greater effect on the ELD of iDCs than highly differentiated ESCC cells.

Airway management using a supraglottic airway device without endotracheal intubation for positive ventilation of anaesthetized rats.

Endotracheal intubation is often necessary for positive pressure ventilation of rats during open thoracic surgery. Since endotracheal intubation in rats is technically difficult and is associated with numerous complications, many techniques using various devices have been described in the scientific literature. In this study, we compared the effectiveness of airway management of a home-made supraglottic airway device (SAD), which is cheap to fabricate and easy to place with that of an endotracheal intubation tube in enflurane-anaesthetized rats. Twenty male Sprague-Dawley rats (200-300 g) were randomly assigned to two equal groups for positive pressure mechanical ventilation using either the SAD or an endotracheal intubation tube. The carotid artery of each rat was cannulated for continuous blood pressure measurements and obtaining blood samples for determination of oxygen tension, carbon dioxide tension, and blood acidity before, during and after SAD placement or endotracheal intubation. Proper placement of the SAD was confirmed by observing chest wall movements that coincided with the operation of the mechanical ventilator. No complications and adverse events were encountered in the rats in which the SAD was placed, during SAD placement and immediate removal, during their mechanical ventilation through the SAD, and one week after SAD removal. From the results of blood gas analyses, we conclude that anaesthetized rats can be successfully ventilated using an SAD for open thoracic surgery.

Blind oral endotracheal intubation of rats using a ventilator to verify correct placement.

Endotracheal intubation in rats is challenging due to difficulties visualizing the epiglottis and vocal cords. No visualization of these structures results in repeated intubation attempts which can cause trauma to the oral cavity and/or oesophagus, and death of the animal due to respiratory failure. Here, we describe a simple blind oral tracheal intubation technique in the rat that decreases the frequency of repeated intubations using an intubation device that comprises a 16 G intravenous catheter and a modified 18 G epidural needle, and a rodent ventilator. The epidural needle is bent in such a way that it curves in conformity with the rat's oral airway in order to direct the catheter into the larynx, and the rodent ventilator is used to verify its correct placement. The first attempt success rate of endotracheal intubation using the blind oral tracheal intubation technique with a rodent ventilator was greater than the first attempt success rate using the blind oral tracheal intubation technique without using a rodent ventilator. Although this method is a simple modification of a previously described method of blind oral endotracheal intubation, our method is easy to learn, inexpensive and does not require specialized equipment.

Association between TNF-alpha promoter polymorphism and Helicobacter pylori cagA subtype infection.

AIMS: To assess the importance of tumour necrosis factor alpha (TNF-alpha) promoter polymorphism in relation to infection with the cytotoxin associated gene A (cagA) subtype of Helicobacter pylori within a dyspeptic Korean population. METHODS: Eighty three patients with gastric disease and 113 healthy controls were studied. The DNA from gastric biopsy specimens was analysed by H pylori specific and cagA specific polymerase chain reaction (PCR). To characterise TNF-alpha polymorphism at positions -308 and -238, PCR based restriction fragment length polymorphism analysis was performed. RESULTS: Helicobacter pylori infection was closely correlated with G to A transition at position -308 of the TNF-alpha promoter when compared with healthy controls (odds ratio (OR), 2.912; 95% confidence interval (CI), 1.082 to 7.836; p = 0.034). Although TNF-alpha -308 polymorphism in patients with H pylori was not significantly different from that in patients without H pylori, the -308A polymorphism was strongly associated with H pylori cagA subtype infection when compared with the polymorphism in cagA negative H pylori infection (OR, 8.757; 95% CI, 1.413 to 54.262; p = 0.019) and healthy controls (OR, 3.683; 95% CI, 1.343 to 10.101; p = 0.011). G to A genetic change at position -238 of the TNF-alpha gene was not significantly associated with H pylori cagA subtype infection. In addition, genetic polymorphisms at both sites of the TNF-alpha promoter in patients with H pylori infection did not correlate with the severity of disease. CONCLUSION: TNF-alpha -308A polymorphism was significantly related to infection with the H pylori cagA subtype in Korean patients with gastric disease.

The -238 tumor necrosis factor-alpha promoter polymorphism is associated with decreased susceptibility to cancers.

We investigated the potential association of tumor necrosis factor-alpha (TNF-alpha) promoter polymorphisms with cancers. The study included 169 patients with gastric cancer, uterine cervical cancer, colorectal cancer, or renal cell carcinoma and 92 healthy controls. The -308 and -238 polymorphisms in the TNF-alpha promoter were analyzed by PCR-restriction fragment length polymorphism (RFLP). The proportion of individuals carrying the TNF-238A allele was significantly lower in the cancer group than in the control group. The odds ratio for cancer in subjects with the TNF-238A allele was 0.25 (95% CI, 0.10-0.64). No association was found between the -308 polymorphism and cancers. These results suggest that the -238A allele has a protective function against cancers.