Cascaded signal amplification strategy for ultra-specific, ultra-sensitive, and visual detection of Shigella flexneri.

Pathogen infections including Shigella flexneri have posed a significant threat to human health for numerous years. Although culturing and qPCR were the gold standards for pathogen detection, time-consuming and instrument-dependent restrict their application in rapid diagnosis and economically less-developed regions. Thus, it is urgently needed to develop rapid, simple, sensitive, accurate, and low-cost detection methods for pathogen detection. In this study, an immunomagnetic beads-recombinase polymerase amplification-CRISPR/Cas12a (IMB-RPA-CRISPR/Cas12a) method was built based on a cascaded signal amplification strategy for ultra-specific, ultra-sensitive, and visual detection of S. flexneri in the laboratory. Firstly, S. flexneri was specifically captured and enriched by IMB (Shigella antibody-coated magnetic beads), and the genomic DNA was released and used as the template in the RPA reaction. Then, the RPA products were mixed with the pre-loaded CRISPR/Cas12a for fluorescence visualization. The results were observed by naked eyes under LED blue light, with a sensitivity of 5 CFU/mL in a time of 70 min. With no specialized equipment or complicated technical requirements, the IMB-RPA-CRISPR/Cas12a diagnostic method can be used for visual, rapid, and simple detection of S. flexneri and can be easily adapted to monitoring other pathogens.

SEMA3B-AS1 suppresses colorectal carcinoma progression by inhibiting Semaphorin 3B-dependent VEGF signaling pathway activation.

Mounting evidence has demonstrated the considerable regulatory effects of long noncoding RNAs (lncRNAs) in the tumorigenesis and progression of various carcinomas. LncRNA Semaphorin 3B (SEMA3B) antisense RNA 1 (SEMA3B-AS1) has been found to be dysregulated in a few carcinomas recently. However, its potential function and mechanism in colorectal carcinoma (CRC) have not yet been examined. Here we show that SEMA3B-AS1 acts as a crucial regulator of CRC progression. We found that SEMA3B-AS1 expression was downregulated in CRC cell lines and tissues. Downregulation of SEMA3B-AS1 was significantly associated with poor survival in CRC patients. Overexpression of SEMA3B-AS1 reduced the cell growth and metastasis of CRC in vivo and in vitro. In addition, SEMA3B-AS1 promoted the expression of its sense-cognate gene SEMA3B, a member of the Semaphorin family (SEMAs), by recruiting EP300 to induce H3K9 acetylation at the SEMA3B promoter. Furthermore, we proved that SEMA3B-AS1 suppressed CRC angiogenesis by affecting the vascular endothelial growth factor signaling pathway activation which was regulated by the SEMA3B-NRP1 axis. Our work unravels a novel mechanism of SEMA3B-AS1 in the inhibition of CRC malignant progression and highlights its probability as a new promising diagnostic marker and therapeutic target for CRC interventions.

Role of noncoding RNAs in liver fibrosis.

Liver fibrosis is a wound-healing response following chronic liver injury caused by hepatitis virus infection, obesity, or excessive alcohol. It is a dynamic and reversible process characterized by the activation of hepatic stellate cells and excess accumulation of extracellular matrix. Advanced fibrosis could lead to cirrhosis and even liver cancer, which has become a significant health burden worldwide. Many studies have revealed that noncoding RNAs (ncRNAs), including microRNAs, long noncoding RNAs and circular RNAs, are involved in the pathogenesis and development of liver fibrosis by regulating signaling pathways including transforming growth factor-ß pathway, phosphatidylinositol 3-kinase/protein kinase B pathway, and Wnt/ß-catenin pathway. NcRNAs in serum or exosomes have been reported to tentatively applied in the diagnosis and staging of liver fibrosis and combined with elastography to improve the accuracy of diagnosis. NcRNAs mimics, ncRNAs in mesenchymal stem cell-derived exosomes, and lipid nanoparticles-encapsulated ncRNAs have become promising therapeutic approaches for the treatment of liver fibrosis. In this review, we update the latest knowledge on ncRNAs in the pathogenesis and progression of liver fibrosis, and discuss the potentials and challenges to use these ncRNAs for diagnosis, staging and treatment of liver fibrosis. All these will help us to develop a comprehensive understanding of the role of ncRNAs in liver fibrosis.

Comparative short-term outcomes of robotic-assisted surgery for older patients with non-small cell lung cancer: a propensity matched study.

To investigate comparative short-term outcomes of robotic-assisted surgery (RAS) versus video-assisted thoracoscopic surgery (VATS) for older non-small cell lung cancer (NSCLC) patients. Patients ≥ 65 years old with stage I-III NSCLC operated with RAS or VATS from 2016 to 2018 were consecutively included. Propensity score-matched (PSM) method was implemented to balance inter-group biases. Totally 376 participants (224 with VATS and 152 with RAS) were included. After PSM, a cohort (144 with VATS and 107 with RAS) was generated with balanced baseline characteristics. RAS was significantly superior over VATS in the majority of perioperative outcomes, such as operating time (120.8 vs. 165.1 min), conversion rate (0.0% vs. 19.4%), and length of stay (8.6 vs. 10.8 days). RAS versus VATS was significantly associated with comparable rates of postoperative complications (OR 0.642, 95% CI 0.311-1.327), except the rate of pneumonia (OR 0.161, 95% CI 0.048-0.544). RAS leads to analogous postoperative complications and seemingly accelerates the recovery time of older NSCLC patients compared with VATS.

[Diagnosis and treatment of small cell neuroendocrine carcinoma of the prostate].

OBJECTIVE: To explore the clinical manifestations, pathological features and treatment of small cell neuroendocrine carcinoma of the prostate (SCPCa). METHODS: We reported 1 case of SCPCa treated in Jinan People's Hospital and analyzed the clinical data on another 57 cases reported in China. RESULTS: Of the total number of cases, 52 (91.2%) had urinary tract symptoms, including 40 cases of urinary tract obstruction symptoms and 53 (91.2%) had abnormal prostatic nodules on digital rectal examination, 17 (29.8%) with PSA > 4 µg/L. The clinical data on 43 (75.4%) of the cases were valuable for the assessment of clinical stages, including 30 cases with confirmed metastases and another 20 with lymph node and distant metastases. Thirty-seven of the cases (64.9%) were reported with immunohistochemical results, 30 (52.6%) with follow-up data (followed up for 1－24 months, with median survival time of 8 months, and a 1-year survival rate of 16.7%), and another 26 with therapeutic information, 14 treated by chemotherapy, with a median survival time of 11 months and a 1-year survival rate of 28.6%, significantly higher than in the non-chemotherapy groups (P<0.01). CONCLUSIONS: Small cell carcinoma of the prostate is highly malignant with poor prognosis. Its clinical symptoms mainly include urinary tract obstruction, its definite diagnosis chiefly depends on histopathology and immunohistochemistry, and its treatment recommended is chemotherapy-based comprehensive protocol.

Anthropogenic influences in a rapidly urbanizing area using linear alkylbenzenes and polycyclic aromatic hydrocarbons as tracers.

Environmental molecular markers can be used to understand the sources, transport, and fate of pollutants. Furthermore, they can also be applied to assess the influences of anthropogenic activities and elucidate urbanization from different perspectives. In this study, the potential of linear alkylbenzenes (LABs) and polycyclic aromatic hydrocarbons (PAHs) as chemical indicators of urbanization was examined first. Overall, the concentrations of LABs and PAHs ranged from 5.49-148 ng/g (mean: 15.6, median: 9.33) and 3.61-4878 ng/g (mean: 181, median: 71.3), respectively. Owing to the different sources and input methods of these two substances in soil, the area-weighted median values for LABs were more suitable to assess the magnitude of contamination on the administrative scale. For PAHs, the average values were more practical. LAB (consumption-induced pollutants) and PAH (production-induced pollutants) concentrations exhibited good correlations with some indices for residential daily life and industrialization, which indicated that soil can be utilized to reveal multidimensional urbanization-environment relationships. Two different patterns, the inverted U-shaped pattern and the upward pattern, were employed to simulate the environment-urbanization relationships in Shenzhen, China, which indicated that raising the standard of living or industrialization had created different soil pollution. The environmental quality demand was more difficult to meet by changing the energy structure than by improving infrastructure.

A Noninvasive Multianalytical Approach for Lung Cancer Diagnosis of Patients with Pulmonary Nodules.

Addressing the high false-positive rate of conventional low-dose computed tomography (LDCT) for lung cancer diagnosis, the efficacy of incorporating blood-based noninvasive testing for assisting practicing clinician's decision making in diagnosis of pulmonary nodules (PNs) is investigated. In this prospective observative study, next generation sequencing- (NGS-) based cell-free DNA (cfDNA) mutation profiling, NGS-based cfDNA methylation profiling, and blood-based protein cancer biomarker testing are performed for patients with PNs, who are diagnosed as high-risk patients through LDCT and subsequently undergo surgical resections, with tissue sections pathologically examined and classified. Using pathological classification as the gold standard, statistical and machine learning methods are used to select molecular markers associated with tissue's malignant classification based on a 98-patient discovery cohort (28 benign and 70 malignant), and to construct an integrative multianalytical model for tissue malignancy prediction. Predictive models based on individual testing platforms have shown varying levels of performance, while their final integrative model produces an area under the receiver operating characteristic curve (AUC) of 0.85. The model's performance is further confirmed on a 29-patient independent validation cohort (14 benign and 15 malignant, with power > 0.90), reproducing AUC of 0.86, which translates to an overall sensitivity of 80% and specificity of 85.7%.

Smoking history and the efficacy of immune checkpoint inhibitors in patients with advanced non-small cell lung cancer: a systematic review and meta-analysis.

BACKGROUND: Some studies imply a strong correlation between smoking history and the efficacy of immune checkpoint inhibitors (ICIs) in patients with advanced non-small cell lung cancer (NSCLC). Hence, a systematic review and meta-analysis was conducted to comprehensively investigate this correlation. METHODS: Three online databases including PubMed, Embase and Cochrane Library were searched. Abstracts and presentations from European Society of Medical Oncology (ESMO) and American Society of Clinical Oncology (ASCO) were also reviewed. The deadline of search was Nov 9, 2019. Randomized clinical trials (RCT) of ICIs that reported hazard ratio (HR) for overall survival (OS) or progressive-free survival (PFS) by the smoking status of NSCLC patients were eligible for our study. We focused on publications issued in English. A random effects model was implemented in the synthesis, and a two-step interaction test was used to investigate the difference of ICIs efficacy among patients with different smoking histories. RESULTS: Twelve RCTs involving 6,497 NACLC patients [5,569 (85.72%) current/former smokers and 928 (114.28%) never smokers] were eligible for our systematic review and meta-analysis. The pooled HRs [95% confidential interval (CI)] of OS and PFS were 0.74 (0.67, 0.81) and 0.72 (0.59, 0.88) respectively for current/former smokers in the experimental group with ICIs versus those in the control group. The pooled HRs (95% CI) of OS and PFS were 0.81 (0.60, 1.08) and 0.92 (0.55, 1.54) respectively for never smokers in the experimental group with ICIs compared with those in the control group. The difference of ICIs efficacy in terms of OS between current/former and never smokers was insignificant [interaction HR (95% CI), 0.77 (0.69, 0.86), I2=25.4%, P_hetero=0.21]. CONCLUSIONS: The efficacy of ICIs in patients with smoking history is seemingly superior over patients without smoking history, but insignificantly. The difference can be explained by several factors such as insufficient sample size of non-smokers, and confounding factors. We suggest that smoking history cannot be recognized as a predictor of immune therapy in advanced NSCLC.

Is the presence of lung injury in COVID-19 an independent risk factor for secondary lung cancer?

The morbidity and mortality of lung cancer are increasing. The Corona Virus Disease 2019 (COVID-19) is caused by novel coronavirus 2019-nCoV-2, leading to subsequent pulmonary interstitial fibrosis with chronic inflammatory changes, e.g., inflammatory factors repeatedly continuously stimulating and attacking the alveolar epithelial cells. Meanwhile, 2019-nCoV-2 can activate PI3K/Akt and ERK signaling pathways, which can play the double roles as both anti-inflammatory and carcinogenic factors. Moreover, hypoxemia may be developed, resulting in the up-regulation of HIF-1 α expression, which can be involved in the occurrence, angiogenesis, invasion and metastasis of lung cancer. Additionally, the immune system in 2019-nCoV-2 infected cases can be suppressed to cause tumor immune evasion. Therefore, we speculate that COVID-19 may be a risk factor of secondary lung cancer.

Serotonin-Mediated Cardiac Analgesia via Ah-Type Baroreceptor Activation Contributes to Silent Angina and Asymptomatic Infarction.

Silent angina is a critical phenomenon in the clinic and is more commonly associated with women patients suffering from myocardial ischemia. Its underlying cause remains mysterious in medicine. With our recent discovery of female-specific Ah-type baroreceptor neurons (BRNs), we hypothesize that cardiac analgesia is due to the direct activation of Ah-type BRNs by elevated levels of circulating serotonin (5-HT) myocardial infarction (MI) patients. Electromyography and the tail-flick reflex were assessed in control and MI-model rats to evaluate 5-HT-mediated BP regulation as well as peripheral and cardiac nociception. 5-HT or a 5-HT receptor agonist was microinjected into the nodose ganglion to confirm the involvement of the afferent pathway of the baroreflex arc. An inward current was observed in identified BRNs by applying a whole-cell patch-clamp technique in conjunction with qRT-PCR to verify the afferent-specific action of 5-HT and the expression of 5-HT receptors. Although the tail-flick reflex and mean arterial pressure were dramatically reduced in female MI rats with elevated serum 5-HT, intrapericardial capsaicin-evoked muscular discharges were significantly inhibited in comparing with those of males, which were mimicked by microinjection of 5-HT or SR57227A into the nodose. Ah-type BRNs displayed robust inward currents at lower concentrations of 5-HT than the C-type or the A-type, with significantly increased expression and cellular distribution of 5-HT3AR but not 5-HT3BR compared to the A- and C-types. Activation of 5-HT3AR in Ah-type BRNs by 5-HT contributes significantly to cardiac analgesia, which may suggest the pathogenic condition that silent angina occurs mainly in female patients.

Comparison of endoscopic submucosal tunneling dissection and thoracoscopic enucleation for the treatment of esophageal submucosal tumors.

BACKGROUND AND AIMS: Endoscopic submucosal tunneling dissection (ESTD) has been proved to be safe and effective for removal of esophageal submucosal tumors (SMTs) and can maintain the mucosal integrity compared with other endoscopic methods. The aim of the study was to estimate the safety and efficacy of ESTD as well as compare its efficacy with thoracoscopic enucleation for esophageal SMTs, which is used increasingly as a minimally invasive approach. METHODS: We retrospectively collected the clinical data of patients with esophageal SMTs <40 mm who underwent ESTD or thoracoscopic enucleation at Nanfang Hospital between January 2008 and August 2016. Epidemiologic data (sex, age), tumor location, tumor size, en bloc resection rate, adverse events, pathologic results, length of postoperative hospital stay, and cost were compared between ESTD and thoracoscopic enucleation. RESULTS: A total of 126 patients were included. A total of 74 patients underwent ESTD, and the other 52 underwent thoracoscopic enucleation. There was no significant difference between the 2 groups in sex, age, tumor size, hospitalization expense, infection, adverse events, and en bloc resection rate (P < .05). However, patients in the ESTD group had a shorter operating time, less estimated blood loss, shorter length of postoperative hospital stay, and lower chest pain level (P < .05). Kaplan-Meier curves for disease-free survival also showed no statistically significant difference between ESTD and thoracoscopic enucleation groups during the median follow-up of 19.5 and 42 months, respectively. CONCLUSIONS: The treatment efficacy was comparable between the ESTD and thoracoscopic enucleation for esophageal SMTs <40 mm. However, there was a significant advantage in the ESTD group for a shorter operating time, reduced postoperative chest pain, and shorter hospitalization.

Application of low dose of FK506 in pancreas transplantation model in Wistar-SD rats.

OBJECTIVE: To probe into application of low dose of FK506(Tacrolimus) in pancreas transplantation. METHODS: Effects of low-dose FK506 (Tacrolimus) in pancreas transplantation with examination of ELISA Electron microscopy and TUNEL by method of random control were studied. RESULTS: Blood glucose concentration in control group is higher than that in treated group A (FK506) and treated group B (CsA) 7 days after transplantation (p < 0.05). Serum C-peptide and insulin concentrations in control group are less than that in treated group A (FK506) and treated group B (CsA) 7 days after transplantation (p < 0.05). Blood glucose, serum C-peptide and insulin concentrations are same as that in control group, group A (FK506) and group B (CsA) (p > 0.05).There are more apoptotic nuclei in control group than that in treated group A (FK506) and treated group B (CsA) (p < 0.05). There is no significant difference between group A (FK506) and treated group B (CsA) in sum of apoptotic nuclei (p > 0.05). There is no significant difference among treated group A (FK506) and treated group B (CsA) in electron microscopy fields. CONCLUSION: Low-dose FK506 applied in pancreas transplantation could not only be effective for immunosuppressive, but also be safe for islet cells of pancreas.

Laparoscopic adrenalectomy in pheochromocytoma: retroperitoneal approach versus transperitoneal approach.

OBJECTIVES: The aim of this study was to evaluate outcomes of transperitoneal and retroperitoneal approaches in laparoscopic adrenalectomy (LA) for pheochromocytoma. METHODS: We retrospectively compared the results of transperitoneal LA (TLA) with that of retroperitoneal LA (RLA) in 99 patients with unilateral pheochromocytoma (<6 cm in diameter). Age of patients ranged from 20 to 63 years (42 men and 57 women; 56 in right and 43 in left). The mean operation time, blood loss, mean hospital stay after operation, fluctuation range of blood pressure, blood pressure after operation, complication, mortalities, and recurrences were considered. RESULTS: There were significant differences between RLA and TLA in terms of duration of surgery 84 ± 17 minutes versus 117 ± 24 minutes (p < 0.05), intraoperative blood loss 200 ± 22.5 ml versus 340 ± 41.7 ml (p < 0.05), mean hospital stay after operation 4.8 ± 1.6 days versus 7.8 ± 2.2 days (p < 0.05), and complication 12.5% versus 6.7% (p < 0.05). There was no significant difference between RLA and TLA in terms of fluctuation range of blood pressure (p > 0.05) and blood pressure after operation (p > 0.05). There were no mortalities or recurrences of tumors in all patients during follow-up. CONCLUSIONS: Compared with TLA, RLA is safer, more quick, and effective for patients with unilateral pheochromocytoma (< 6 cm in diameter).

Mechanisms of diabetes mellitus induced with FK506 in SD rats models.

BACKGROUND: Tacrolimus causes post-transplant diabetes mellitus, however the pathogenetic mechanisms remain controversial. In this study we probed into the mechanisms of tacrolimus-induced diabetes mellitus in rats. METHODS: Glucose levels were determined on whole blood samples using a glucose oxidase method. Levels of serum insulin and C-peptide were measured with ELISA. Histological damage of ultra-structure and apoptosis of beta cells of the pancreas were assayed with electric microscope and tunnel methods respectively.--Ultra-structure were assayed with electric microscope and apoptosis of beta cells of the pancreas were assayed with tunnel methods. Immunohistochemistry was utilized to detect the sum of insulin receptors of hepatic cells. RESULTS: Compared to control group, insulin and C peptide levels in serum decreased in rats of diabetes mellitus models induced with FK506(P<0.05). Compared to the control group, the sum of apoptosis body in pancreatic islets increased in rats of diabetes mellitus models induced with FK506 (P<0.05). Compared to the control group, electron microscopy showed cytoplasm swelling and vacuolization, and marked decrease or absence of dense-core secretory granules in beta cells in rats with diabetes mellitus induced with FK506.Compared to the control group, expression of insulin receptor of hepatic cell decreased in rats of diabetes mellitus models induced with FK506 (P<0.05). CONCLUSION: Pathogenetic mechanisms of rats of diabetes mellitus models induced with FK506 including reduction of secretion of insulin in beta cells of pancreatic islets, damages of ultra-structure of beta cells of pancreatic islets, increasing of apoptosis of beta cells of pancreatic islets and decreasing of expression of insulin receptors in hepatic cells.