Integrated pan-cancer analysis and experimental verification of the roles of tropomyosin 4 in gastric cancer.

Objective: To investigate the function of tropomyosin 4 (TPM4) using pan-cancer data, especially in gastric cancer (GC), using comprehensive bioinformatics analysis and molecular experiments. Methods: We used UCSC Xena, The Cancer Genome Atlas (TCGA), Genotype-Tissue Expression Project (GTEx), TIMER2.0, GEPIA, cBioPortal, Xiantao tool, and UALCAN websites and databases for the extraction of pan-cancer data on TPM4. TPM4 expression was investigated with respect to prognosis, genetic alterations, epigenetic alterations, and immune infiltration. RNA22, miRWalk, miRDB, Starbase 2.0, and Cytoscape were used for identifying and constructing the regulatory networks of lncRNAs, miRNAs, and TPM4 in GC. Data from GSCALite, drug bank databases, and Connectivity Map (CMap) were used to analyze the sensitivity of drugs dependent on TPM4 expression. Gene Ontology (GO), enrichment analyses of the Kyoto Encyclopedia of Genes and Genomes (KEGG), wound healing assays, and (Matrigel) transwell experiments were used to investigate the biological functions of TPM4 in GC. Result: The findings of the comprehensive pan-cancer analysis revealed that TPM4 has a certain diagnostic and prognosis value in most cancers. Alterations in the expression of TPM4, including duplications and deep mutations, and epigenetic alterations revealed that TPM4 expression is related to the expression of DNA methylation inhibitors and RNA methylation regulators at high concentrations. Besides, TPM4 expression was found to correlate with immune cell infiltration, immune checkpoint (ICP) gene expression, the tumor mutational burden (TMB), and microsatellite instability (MSI). Neoantigens (NEO) were also found to influence its response to immunotherapy. A lncRNA-miRNA -TPM4 network was found to regulate GC development and progression. TPM4 expression was related to docetaxel,5-fluorouracil, and eight small molecular targeted drugs sensitivity. Gene function enrichment analyses revealed that genes that were co-expressed with TPM4 were enriched within the extracellular matrix (ECM)-related pathways. Wound-healing and (Matrigel) transwell assays revealed that TPM4 promotes cell migration and invasion. TPM4, as an oncogene, plays a biological role, perhaps via ECM remodeling in GC. Conclusions: TPM4 is a prospective marker for the diagnosis, treatment outcome, immunology, chemotherapy, and small molecular drugs targeted for pan-cancer treatment, including GC treatment. The lncRNA-miRNA-TPM4network regulates the mechanism underlying GC progression. TPM4 may facilitate the invasion and migration of GC cells, possibly through ECM remodeling.

The role of crm-1 in ionizing radiation-induced nervous system dysfunction in Caenorhabditis elegans.

Ionizing radiation can cause changes in nervous system function. However, the underlying mechanism remains unclear. In this study, Caenorhabditis elegans (C. elegans) was irradiated with 75 Gy of 60Co whole-body γ radiation. Behavioral indicators (head thrashes, touch avoidance, and foraging), and the development of dopaminergic neurons related to behavioral function, were evaluated to assess the effects of ionizing radiation on nervous system function in C. elegans. Various behaviors were impaired after whole-body irradiation and degeneration of dopamine neurons was observed. This suggests that 75 Gy of γ radiation is sufficient to induce nervous system dysfunction. The genes nhr-76 and crm-1, which are reported to be related to nervous system function in human and mouse, were screened by transcriptome sequencing and bioinformatics analysis after irradiation or sham irradiation. The expression levels of these two genes were increased after radiation. Next, RNAi technology was used to inhibit the expression of crm-1, a gene whose homologs are associated with motor neuron development in other species. Downregulation of crm-1 expression effectively alleviated the deleterious effects of ionizing radiation on head thrashes and touch avoidance. It was also found that the expression level of crm-1 was regulated by the nuclear receptor gene nhr-76. The results of this study suggest that knocking down the expression level of nhr-76 can reduce the expression level of crm-1, while down-regulating the expression level of crm-1 can alleviate behavioral disorders induced by ionizing radiation. Therefore, inhibition of crm-1 may be of interest as a potential therapeutic target for ionizing radiation-induced neurological dysfunction.

[Potentilla anserina polysaccharide alleviates the damage of mouse glomerular mesangial cells induced by high glucose via promoting the expression of circ-AKT3].

Objective To explore the effect of Potentilla anserina polysaccharide on the injury of glomerular mesangial cells induced by high glucose and its possible mechanism. Methods High glucose was used to induce SV40 MES 13 mouse glomerular mesangial cells to establish a cell injury model, and 0.1, 0.3, 0.6 mg/mL Potentilla anserina polysaccharides were added to treat the cells. pcDNA and pcDNA-circ-AKT3 were respectively transfected into mesangial cells induced by high glucose. si-NC and si-circ-AKT3 were transfected into glomerular mesangial cells induced by high glucose and Potentilla anserina polysaccharide was added to the cells. The levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and malondialedhyde (MDA) were tested according to the kit. The levels of IL-6, IL-18 and monocyte chemoattractant protein-1 (MCP-1) were detected by ELISA. Flow cytometry was used to detect the apoptosis rate. The real time quantitative PCR was used to detect the expression of circ-AKT3 mRNA. Results In glomerular mesangial cells induced by high glucose, the levels of SOD and GSH-Px were decreased, but the levels of MDA, IL-6, IL-18, and MCP-1 were increased, along with the increased rate of apoptosis, and decreased the expression level of circ-AKT3. After Potentilla anserina polysaccharide treatment of glomerular mesangial cells induced by high glucose, the levels of SOD and GSH-Px were increased whereas the levels of MDA, IL-6, IL-18, and MCP-1 were decreased, together with decreased apoptosis rate, and increased expression level of circ-AKT3. The difference was significant among different concentration groups. pcDNA-circ-AKT3 was transfected into glomerular mesangial cells induced by high glucose, followed by the increased levels of SOD and GSH-Px, with the decreased levels of MDA, IL-6, IL-18 and MCP-1. The apoptosis rate was also reported decreased. Interference of circ-AKT3 expression, on the other hand, can restore the protective effect of Potentilla anserina polysaccharide on high glucose induced glomerular mesangial cell injury. Conclusion Potentilla anserina polysaccharide can alleviate the damage of glomerular mesangial cells induced by high glucose by increasing the expression of circ-AKT3.

Protective Effects of Polydatin on Reproductive Injury Induced by Ionizing Radiation.

The reproductive system is vulnerable to ionizing radiation, which is a hot research topic at present. We tested the effect of polydatin on spermatocytes(GC-1 cells) after X-ray irradiation. The reproductive damage model of C.elegans was established by 60Coγ-ray, and the protective effect of polydatin on reproductive damage caused by ionizing radiation was evaluated. We quantified the ROS levels of GC-1 cells and C.elegans after irradiation with polydatin and evaluated the anti-apoptosis effect of polydatin at proper concentration. Differential genes of C.elegans reproductive damage were screened out from transcriptome sequencing results and comparable GEO datasets. It was proved that 100µM polydatin significantly reduced the apoptosis of GC-1 cells induced by 2 Gy X-ray. In addition, the longevity, reproductive capacity, germ cell apoptosis and spawning and hatching capacity of polydatin were tested. The results showed that 100 µM polydatin content significantly increased the influence of 50 Gy 60Coγ-ray on reproductive capacity of C.elegans. Quantitative analysis of mRNA and protein levels of apoptosis-related genes and reproductive-related genes by qRT-PCR and Western blotcon firmed that polydatin with appropriate dosage had good protective effects on reproductive damage caused by radiation, which laid a foundation for the application research of polydatin in radiation protection.

Predictive performances of 6 data mining techniques for obstructive sleep apnea-hypopnea syndrome.

PURPOSE: This study compared the effects of 6 types of obstructive sleep apnea-hypopnea syndrome (OSAHS) prediction models to develop a reference for selecting OSAHS data mining tools in clinical practice. METHODS: This cross-sectional study included 401 cases. They were randomly divided into 2 groups: training (70%) and testing (30%). Logistic regression, a Bayesian network, an artificial neural network, a support vector learning machine, C5.0, and a classification and regression tree were each adopted to establish 6 prediction models. After training, the 6 models were used to test the remaining samples and calculate the correct and error rates of each model. RESULTS: Twenty-one input variables for which the difference between the patient and nonpatient groups was statistically significant were considered. The models found the abdominal circumference, neck circumference, and nocturia ≥2 per night to be the most important variables. The support vector machine, neural network, and C5.0 models performed better than the classification and regression tree, Bayesian network, and logistic regression models. CONCLUSIONS: In terms of predicting the risk of OSAHS, the support vector machine, neural network, and C5.0 were superior to the classification and regression tree, Bayesian network, and logistic regression models. However, such results were obtained based on the data of a single center, so they need to be further validated by other institutions.

Knockdown of GPRC5B alleviates the high glucose-induced inflammation and extracellular matrix deposition of podocyte through inhibiting NF-κB pathway.

BACKGROUND: Diabetes is a serious disease that could greatly increase the risk of cardiovascular complications, whereas the underlying pathology of DN is still unknown. GPRC5B is a member of the RAIG subfamily of type 3 (family C) GPCR, and its role in DN is still unclear. OBJECTIVE: To unveil the role of GPRC5B in diabetic nephropathy (DN) progression and investigate the potential signaling pathway. MATERIALS AND METHODS: Podocytes were stimulated with high glucose and expression of GPRC5B was analyzed by qPCR and western blot. Then the level of GPRC5B was depleted by siRNA transfection and inflammatory cytokine level was monitored by ELISA assay. The ECM depostion and the activation of NF-κB pathway were detected by Immunoblot. RESULTS: We investigated the possible role of GPRC5B in the pathology of diabetic nephropathy. We found GPRC5B was highly expressed in high glocuse (HG) induced podocytes. The depletion of GPRC5B inhibited HG induced cell inflammation. In addition, the ablation of GPRC5B suppressed the HG induced ECM deposition. We further found GPRC5B could alleviate the inflammation and extracellular matrix deposition of HG-induced podocytes through NF-κB pathway. CONCLUSION: We therefore thought GPRC5B could serve as a promising target for the treatment of diabetic nephropathy. G-protein-coupled receptors.

MUC1 Promotes Mesangial Cell Proliferation and Kidney Fibrosis in Diabetic Nephropathy Through Activating STAT and ß-Catenin Signal Pathway.

Diabetic nephropathy (DN) is a complication of diabetes, which leads to most end-stage kidney diseases and threatens health of patients. Mucin 1 (MUC1) is a heterodimeric oncoprotein, which is abnormally expressed in tumors and hematologic diseases. The aim of this study is to clarify the mechanism and role of MUC1 in DN. The mesangial cells (MCs) suffered from high glucose (HG) treatment to mimic DN in vitro. The cell proliferation was detected by Cell Counting Kit-8 assay and 5-ethynyl-2-deoxyuridine (EdU) staining assay. The expression of MUC1 and fibrosis markers: fibronectin, collagen I, and collagen IV were assessed by western blot. In this study, we demonstrated that HG treatment induced MUC1 expression in MCs. With knockdown of MUC1 or overexpressed MUC1 in MCs, the results indicated that knockdown of MUC1 inhibited MCs proliferation and reduced kidney fibrosis markers expression, including fibronectin, collagen I, and collagen IV, whereas overexpression of MUC1 led to opposite results. Mechanically, MUC1 activated signal transducers and activators of transcription (STAT) and ß-catenin signal pathway. After added AG490 (STAT inhibitor) or FH535 (ß-catenin inhibitor), blocking STAT3 and ß-catenin signal pathway attenuated MUC1-induced cell proliferation and fibronectin production in MCs. Finally, knockdown of MUC1 attenuated DN-induced kidney fibrosis in db/db mice. Therapeutic target for DN. In conclusion, MUC1 promotes MCs proliferation and kidney fibrosis in DN through activating STAT and ß-catenin signal pathway, which can help to provide a novel therapeutic target for DN.

Bridging "Office-Based Care" With the "Virtual Practice Care Model": Evolving Care for Chronic Kidney Disease Patients in the COVID-19 Pandemic-And Beyond.

Since the outbreak of the coronavirus epidemic, the "virtual" telemedicine has become a critical substitute for patient-provider interactions. However, virtual encounters often face challenges in the care of patients in high-risk categories such as chronic kidney disease (CKD) patients. In this study, we explore the patient's satisfaction and the practical effects of a newly established telemedicine program on CKD patients' care during the COVID-19 pandemic. Based on a prior version of an online patient care platform established in 2017, we developed a customized and improved online telemedicine program designed to specifically address the challenges emerging from the pandemic. This included an online, smart phone-based strategy for triage and medical care delivery and psychological support. We invited a total of 278 CKD patients to join the new platform during the pandemic. The subjects in group A were patients utilizing our old online CKD system and were historical users registered at least 3 months before the pandemic. A pilot survey interrogating medical and psychological conditions was conducted. Feedback on the program as well as a psychological assessment were collected after 1 month. In total, 181 patients showed active responses to the program, with 289 person-time medical consultations occurring during the study. The virtual care program provided a rapid triage for 17% (30 out of 181) patients, with timely referral to in-patient medical encounters for their worsening medical conditions or severe psychological problems. Nearly all patients (97.4%) believed the program was helpful. The number of symptoms (OR 1.309, 95%CI 1.113-1.541; P = 0.001) and being enrolled during the pandemic (OR 3.939, 95% CI 1.174-13.221; P = 0.026) were associated with high stress. During the follow-up, the high-stress CKD group at baseline showed a significant decrease in avoidance score (6.9 ± 4.7 vs. 9.8 ± 1.9, P = 0.015). In conclusion, during the pandemic, we established an online telemedicine care program for CKD patients that provides a rapid triage function, effective CKD disease management, and potentially essential psychological support.

Effects of 60Co γ Irradiation on the Reproductive Function of Caenorhabditis elegans.

The effects of ionizing radiation on the reproductive system have always been a matter of great interest. Both artificial and naturally occurring ionizing radiation can directly or indirectly affect the reproductive system via the introduction of DNA single-strand and double-strand breaks, the excitation of water molecules, and the generation of free radicals. In order to quantitatively investigate the effects of ionizing radiation on reproductive function, 60Co γ irradiation was applied on a model organism, Caenorhabditis elegans (C. elegans). The egg-laying and embryo-hatching activities were observed for the parent (F0) and the first 2 progeny (F1 and F2) generations. The incidence rate of ovipositor malformation was also recorded. Acridine orange was used to detect the number of apoptotic germ cells. With the above metrics, the effects of 60Co γ irradiation on the reproductive function of C. elegans were systematically evaluated. The results showed that the postirradiation egg-laying and embryo-hatching activities of the F0 generation were increasingly suppressed by increasing doses of 60Co γ irradiation. Those of the F1 generation showed a trend toward recovery although also suppressed by the radiation to the F0 generation compared with the control. Those activities were restored to normal or near-normal levels for the F2 generation. The incidence rate of ovipositor malformation was greatly increased by 60Co γ irradiation according to radiation doses. Gamma irradiation by 60Co also substantially induced germ cell apoptosis, and the apoptosis rate increased with increasing radiation doses. Therefore, 60Co γ irradiation affects the reproductive function of C. elegans. The suppression on its reproductive function increases with increasing radiation doses. The reproductive functions of progeny generations are also affected and weakened.

Effect of sulfated galactan from Porphyra haitanensis on H2O2-induced premature senescence in WI-38 cells.

Porphyran sulfated galactan extracted from red algae Porphyra haitanensis is a sulfated polysaccharide, which possesses excellent activities. In the present study, WI-38 cells were treated with H2O2 to induce premature senescence and then the protection of porphyran against aging in vitro and associated molecular mechanisms were investigated. The protection occurred in a dose-dependent manner, offering an optimal efficacy starting at 10µg/mL. The proportion of SA-ß-gal positive cells in porphyran group decreases from 53% to 23% in the cultures at 30 PDs. Porphyran has been detected specifically reducing SAHF-like foci formation in senescent cells. In addition, porphyran significantly affected the p53-p21 pathways in H2O2-treated WI-38 cells. Our data suggest the promising role of porphyran as an attractive and bio-safe agent with the potential to retard senescence and attenuate senescence-related diseases.

A Nomogram for Predicting the Likelihood of Obstructive Sleep Apnea to Reduce the Unnecessary Polysomnography Examinations.

BACKGROUND: The currently available polysomnography (PSG) equipments and operating personnel are facing increasing pressure, such situation may result in the problem that a large number of obstructive sleep apnea (OSA) patients cannot receive timely diagnosis and treatment, we sought to develop a nomogram quantifying the risk of OSA for a better decision of using PSG, based on the clinical syndromes and the demographic and anthropometric characteristics. METHODS: The nomogram was constructed through an ordinal logistic regression procedure. Predictive accuracy and performance characteristics were assessed with the area under the curve (AUC) of the receiver operating characteristics and calibration plots, respectively. Decision curve analyses were applied to assess the net benefit of the nomogram. RESULTS: Among the 401 patients, 73 (18.2%) were diagnosed and grouped as the none OSA (apnea-hypopnea index [AHI] <5), 67 (16.7%) the mild OSA (5 ≤ AHI < 15), 82 (20.4%) the moderate OSA (15 ≤ AHI < 30), and 179 (44.6%) the severe OSA (AHI ≥ 30). The multivariable analysis suggested the significant factors were duration of disease, smoking status, difficulty of falling asleep, lack of energy, and waist circumference. A nomogram was created for the prediction of OSA using these clinical parameters and was internally validated using bootstrapping method. The discrimination accuracies of the nomogram for any OSA, moderate-severe OSA, and severe OSA were 83.8%, 79.9%, and 80.5%, respectively, which indicated good calibration. Decision curve analysis showed that using nomogram could reduce the unnecessary polysomnography (PSG) by 10% without increasing the false negatives. CONCLUSIONS: The established clinical nomogram provides high accuracy in predicting the individual risk of OSA. This tool may help physicians better make decisions on PSG arrangement for the patients referred to sleep centers.

Synthesis of tetracycline-imprinted polymer microspheres by reversible addition-fragmentation chain-transfer precipitation polymerization using polyethylene glycol as a coporogen.

Tetracycline (TC)-imprinted microspheres have been synthesized by reversible addition-fragmentation chain-transfer precipitation polymerization using PEG as a coporogen. In the synthesis, methacrylic acid and ethylene dimethacrylate were used as the functional monomer and cross-linker, respectively. 2,2'-Azobisisobutyronitrile was the initiator, and cumyl dithiobenzoate was the chain-transfer reagent. Although monodispersed microspheres were obtained using acetonitrile as porogen, the particles cannot be used in the column extraction because of the high backpressure. To increase the porosity of the material, PEG was introduced as a coporogen. The influence of the molecular weight and concentration of PEG on the morphology, binding affinity, and porosity of the molecularly imprinted polymers (MIPs) have been studied. The results demonstrated that PEG as a macroporogen increased the porosity of the polymers. Meanwhile, the column backpressure was reduced using the MIPs with higher porosity. The binding affinity of the MIPs was increased when a low concentration of PEG was employed, while it was decreased when the ratio of PEG 12,000/monomers was >0.8%. Under the optimized conditions, TC-imprinted microspheres with good selectivity and size uniformity have been obtained, which facilitates its application in the column extraction for TC determinations.

[Relationship of continuous positive airway pressure and pulmonary function in patients with obstructive sleep apnea-hypopnea syndrome].

OBJECTIVE: To explore the relationship of continuous positive airway pressure and pulmonary function in patients with obstructive sleep apnea hypopnea syndrome (OSAHS), and formulate the prediction equation for the effective therapeutic pressure. METHODS: In a retrospective cross-sectional study of 48 patients with established OSAHS, all the patients were carefully examined for their medical history, and overnight sleep monitoring was carried out to measure the sleep apnea-hypopnea index, mean oxygen saturation, minimum oxygen saturation, and oxygen drop index. The data of manual pressure titration (effective pressure) and pulmonary function tests (tidal volume, one second forced expiratory volume, central airway resistance, and peripheral airway resistance were collected for multiple linear regression analysis. RESULTS: The effective therapeutic pressure was not correlated with the indices of the pulmonary functions in the patients, but showed correlations with the neck circumference, abdominal circumference, apnea-hypopnea index, mean oxygen saturation, least oxygen saturation, and oxygen desaturation index. Multiple linear regression (α=0.05 test level) identified only oxygen desaturation index (P=0.012) and mean oxygen saturation (P=0.036) as the dependent variables of the effective therapeutic pressure. Linear regression analysis showed that the effective therapeutic pressure had a linear relationship with the oxygen drop index and mean oxygen saturation, and was inversely correlated with the mean oxygen saturation. CONCLUSIONS: In patients with OSAHS, the pulmonary function indices are not correlated with the effective therapeutic pressure. In the absence of manual pressure titration, the effective therapeutic pressure can be predicted using the prediction equation: effective therapy pressure = 24.262+0.044×oxygen desaturation index -0.19×average oxygen saturation.