Search

Background: Liver Hepatocellular Carcinoma (LIHC) is characterized by high malignancy, poor prognosis, and high recurrence rate worldwide. The role of ferroptosis in tumorigenesis and progression has been confirmed in previous studies. However, the multi-omics analysis in liver cancer of ferroptosis-markers RPL8 remains to be elucidated. Methods: In this analysis, the RPL8 mRNA expression was analyzed via the GEPIA, TIMER and UALCAN databases. In addition, we verified the mRNA expression of RPL8 by qRT-PCR experiment. The Kaplan-Meier plotter, UALCAN, TCGAportal and HPA databases were applied to evaluate RPL8 on prognosis and clinicopathological parameters. Moreover, we used TIMER and Kaplan-Meier plotter to analyze the correlation of RPL8 to immune cell infiltration and immune cell type markers to prognosis. In addition, networks and function enrichment between RPL8 coexpression genes were analyzed by GeneMANIA, cBioportal and Metascape databases. What’s more, we used FerrDb and GEPIA databases to analyze the correlation of 23 Ferroptosis-related genes with RPL8. Results: The mRNA expression of RPL8 was over-expressed in multiple cancers. In addition, transcription and translation levels of RPL8 in LIHC were significantly higher than normal tissues. Furthermore, higher expression of RPL8 was closely related to shorter OS in LIHC patients. The analysis of Kaplan-Meier plotter proved that RPL8 expression was related to stage, Sorafenib treatment, alcohol consumption and hepatitis virus. Moreover, the results showed that the methylation expression level of RPL8 was significantly associated with age, gender, grade, stage and TP53 mutation of LIHC. RPL8 and its co-expression genes were primarily involved in liver regeneration and immune system process. Immune infiltration analysis showed the RPL8 expression had positively correlated with immune cells and immune subtypes in LIHC. Furthermore, qRT-PCR experiment validated the expression difference of RPL8 in liver cancer. Conclusion: Our findings elucidated that ferroptosis-markers RPL8 may play an important role in prognosis, and significantly correlate with ferroptosis-related genes, it also revealed the potential of RPL8 as a novel therapeutic target for LIHC treatment and prognosis assessment.

Subclinical rejection (SCR) is a common injury in protocol biopsy after pediatric liver transplantation (pLT), but its effect on the recipient is not clearly understood. We herein investigated the incidence and risk factors involved in SCR and analyzed the relationship between SCR and allograft fibrosis (AF).We retrospectively reviewed the biopsy results from 507 children between May 2013 and May 2019, and 352 patients underwent protocol biopsy two years after pLT, 203 underwent protocol biopsy five years after pLT, and 48 underwent protocol biopsy both two and five years after pLT.The incidence of SCR in the five-year group was higher than that in the two-year group (20.2%vs.13.4%, respectively, P = 0.033). The number of patients with mild and moderate SCR in the 5-year group was also higher than that in the 2-year group(P = 0.039). Logistic regression analysis showed that acute rejection before liver biopsy and cadaver liver transplantation (DDLT) were independent risk factors for SCR in the two groups, and that the incidence and severity of AF in protocol biopsies at both periods in the SCR group were higher than those in the non-SCR group (P0.05).The incidence and severity of SCR increased with the prolongation of protocol biopsy time. We postulate that acute rejection and DDLT are independent risk factors for SCR after transplantation. As the occurrence of SCR also augmented the incidence and severity of AF. This article is protected by copyright. All rights reserved.

Yan Liu,1 Zhenglu Wang,2 Lin Zhao1 1Department of Obstetrics, Tianjin First Central Hospital, Nankai University, Tianjin, 300192, People’s Republic of China; 2Biobank, Tianjin First Central Hospital, Nankai University, Tianjin, 300192, People’s Republic of ChinaCorrespondence: Lin ZhaoDepartment of Obstetrics, Tianjin First Central Hospital, Nankai University, No. 24 Fukang Road, Nankai District, Tianjin, 300192, People’s Republic of ChinaTel +86 22-85358245Email 5020200076@nankai.edu.cnBackground: Hypertensive disorders of pregnancy affect approximately 5– 10% of all pregnancies, and this study aims to identify potential diagnostic signatures.Methods: We downloaded the mRNA profiles of GSE75010 (placenta samples) and GSE48424 (blood samples) datasets with or without hypertension in pregnancy from the Gene Expression Omnibus database. Differential expression analysis was performed on the placenta samples using limma package of R language. GO terms and KEGG pathways enrichment analyses were performed on the placenta samples by the clusterProfiler package of R language. Infiltrating immune cell proportion of the placenta samples was evaluated using CIBERSORT software. The key genes involved in hypertension in pregnancy were screened from protein–protein interaction (PPI) network constructed based on the differentially expressed genes (DEGs). The logistic regression model was constructed by the glm package of R language, and receiver operating characteristic (ROC) curve was plotted to determine the accuracy of the model.Results: For the placenta samples, a total of 104 DEGs were identified, and 39 GO terms and 7 KEGG pathways were significantly enriched based on these 104 genes. Furthermore, the analysis of infiltrating immune cells indicated that the difference in the amount of immune cells might be the potential cause of hypertension in pregnancy. The logistic regression model was constructed based on three optimal genes (LEP, PRL and IGFBP1) screened from PPI network and could efficiently separate patients with hypertension in pregnancy from healthy subjects.Conclusion: A predictive model based on three potential genes LEP, PRL and IGFBP1 was obtained, suggesting that these genes might be potential diagnostic signatures for hypertension in pregnancy.Keywords: hypertension in pregnancy, diagnostic signature, GO terms and KEGG pathways analyses, PPI network, logistic regression model

Objective To explore gestational diabetes mellitus (GDM) diagnostic markers and establish the predictive model of GDM. Methods We downloaded the DNA methylation data of GSE70453 and GSE102177 from the Gene Expression Omnibus database. Epigenome-wide association study (EWAS) was performed to analyze the relationship between cytosine-phosphate-guanine (CpG) methylation and GDM. And then the logistic regression models were constructed, with the β-values of CpG sites as predictor variable and the GDM occurrence as binary outcome variable. Data from GSE70453 served as training sets and data from GSE102177 served as verification sets. Results The EWAS and overlap analysis identified nine-shared significant CpGs in the two DNA methylation data sets. Remarkably, these nine CpGs were differently methylated in GDM samples compared to their matched normal specimens, among which five fully methylated CpGs were finally selected. Importantly, we established a binary logistic regression model based on the above five CpGs, in which cg11169102, cg21179618 and cg21620107 were critical. Hence, we further built a logistic regression model by using the three CpGs and found that the area under the curve was 0.8209. The validation of the model by using the verification sets indicated the area under the curve was 0.8519. Conclusions We identified potential CpG biomarkers for the diagnosis of gestational diabetes mellitus patients through using EWAS and Logistic regression models in combination.

Although the significance of the coupled Fe- and N- cycling processes on biogeochemical transformation in riparian wetlands is well-known, the regulation associated with the changes on the microbiotas during different hydrological regimes remains unclear. This study performed field investigations on the bacterial community compositions (BCC) and specific genera associated to Fe- and N- cycling in the rhizosphere soil and sediments in a riparian wetland in Poyang lake, China. The predominant phyla Proteobacteria, Acidobacteria, and Nitrospirae from all the samples remarkably decreased after long-term continuous flooding, while Actinobacteria, Firmicutes and Bacteroidetes were enriched. For the family level, the relative abundances of iron-oxidizing bacteria (FeOB) Gallionellaceae, and N fixing bacteria Nitrospiraceae and Bradyrhizobiaceae significantly declined upon the long-term flooding and then increased with dewatering, which were consistent with the functional genes sequencing analysis. In which, the Bradyrhizobiaceae (RA 2.0 %-34.6 %) was the dominant nirS denitrifier and potential iron-reducing bacteria (FeRB), Sideroxydans lithotrophicus was one of the dominant FeOB (RA 1.7 %-23 %), which was also identified to be the nirS dentrifier (RA 0.2 %-4.3 %). The absolute quantification of the functional genes levels including nirS, nirK, FeRB (Geobacter spp.) showed their significant increases by 3-7 times upon desiccation compared to that under post-CF. The PCA and RDA results indicated the linkage between redox changes of N and Fe during inundation mediated by FeRB, NOB, and FeOB, which were closely related to hydrochemical indices NO

Background Household air pollution (HAP) from residential combustion considerably affects human health in rural China. Large-scale population migration and rural lifestyle changes during the Spring Festival are supposed to change the household air pollution and health risks; however, limited field study has determined its impacts on HAP and short-term health outcomes. Methods A field study was conducted in rural areas of Southern China before and during the Spring Festival to explore the associations between HAP and blood pressure considering different factors such as cooking fuel, heating fuel, and smoking. Stationary real-time PM2.5 monitors were used to measure PM2.5 concentrations of the kitchen, living room, and yard of 156 randomly selected households. Personal exposure to PM2.5 was calculated based on the results of stationary samplers and corresponding time local residents spent in different microenvironments, and one adult resident was recruited of each family for the blood pressure measurement. Results Both personal exposure to PM2.5 and blood pressures of local residents increased during Spring Festival compared to the days before the holiday. Based on generalized linear model coupled with dominance analysis approach, it was found that personal PM2.5 exposure was positively associated with the factors of population size and the types of cooking and heating fuels with the relative contributions of approximately 82%, and systolic blood pressure (SBP, 100-120 mmHg as normal range for adults) was positively and significantly associated with personal PM2.5 exposures with the relative contribution of 11%. Conclusion The findings in this study demonstrated that Spring Festival can give rise to increase of HAP and hypertension risks, also related to tremendous solid fuel use, suggesting further policy making on promoting cleaner energy in rural areas and more attention on large population migration during national holidays.

It was well recognized that the pollutant emissions from real-world residential coal combustion contributed significantly to air pollution and human health, resulting in an urgent need for a better understanding of the pollutant emission factors (EFs) of residential coal combustion. However, field-based researches on residential coal combustion emissions are still limited at this stage. In this study, based on carbon balance method, emission factors (EFs) of CO2 and a series of pollutants, including gaseous pollutants (CO, NOX, CH4, and SO2), particulate matters (PM2.5 and PM10), elemental carbon (EC), organic carbon (OC), and polycyclic aromatic hydrocarbons (PAHs) from residential honeycomb briquette (HB) and coal chunk (CC) combustions were measured in real-world cooking stoves. As a result, apart from SO2, the burning of HB produced much less pollutants than CC. The median of emission reduction ratios estimated from Monte Carlo simulation ranged from 2.4% (SO2) to 98.8% (EC) by replacing CC with HB in residential combustion in Shanxi province in 2012. EFs of CO, PM2.5 and PAHs derived from the present field measurement were around 1–50 times higher than those from laboratory tests; furthermore, larger variations were found in our field measurement compared to that in the laboratory studies. This result called for more field measurements to reduce uncertainties of emission inventory estimations. As for the PAH emissions, particle phase PAHs dominated in both HB and CC burning and PAH emissions from HB burning had lower toxicity than CC combustion. Hence, HB is an ideal substituted fuel compared with CC. Previous studies suggested that PAH isomer ratios of FLA/(FLA+PYR) and IcdP/(IcdP+BghiP) above 0.5 represented source of coal combustion, which did not fit with our study well, indicating PAH isomer ratios might not be valid enough for source apportionment.

Many studies have reported the occurrence of microplastics in live shellfish intended for human consumption. However, far fewer studies have been conducted on dried shellfish from supermarkets or fishery markets. In this study, the characteristics of microplastics in six kinds of dried shellfish products following different cooking treatments were investigated. Dietary exposure to microplastics in dried shellfish was estimated using the consumption rate of seafood among different age groups. Microplastics were detected in all the uncooked, dried shellfish products, ranging from 0.3 to 4.2 items/g. Fibres accounted for more than 80% of microplastics in razor clams, winkles, and scallops. The proportion of microplastics smaller than 1 mm in size ranged from 57.1% to 89.7% of the total microplastics found in dried shellfish. The polymer types included polyethylene terephthalate (PET), rayon, polyester, nylon, polypropylene (PP), cellophane (CP), and polyethylene (PE). Principal component analysis (PCA) showed that the sizes and shapes of microplastics in scallops were more susceptible to alteration by different cooking methods. Steaming and frying significantly reduced the abundance of microplastics in razor clams. In addition, significantly fewer microplastics were found in scallop products after boiling and steaming than were found in fried scallop products. The estimated dietary intake of microplastics for infants was the highest among the age groups considered (3.05 items/kg(bw)/day). Accordingly, frying was suggested for cooking mussels, boiling for clams and winkles, and steaming for scallops. Combining risks from ingesting plastics and plastic additives, steaming is suggested as the best method to cook shellfish.

Transplant oncology is a newly emerging discipline integrating oncology, transplant medicine, and surgery and has brought malignancy treatment into a new era via transplantation. In this context, obtaining a drug with both immunosuppressive and antitumor effects can take into account the dual needs of preventing both transplant rejection and tumor recurrence in liver transplantation patients with malignancies. Capecitabine (CAP), a classic antitumor drug, has been shown to induce reactive oxygen species (ROS) production and apoptosis in tumor cells. Meanwhile, we have demonstrated that CAP can induce ROS production and apoptosis in T cells to exert immunosuppressive effects, but its underlying molecular mechanism is still unclear. In this study, metronomic doses of CAP were administered to normal mice by gavage, and the spleen was selected for quantitative proteomic and phosphoproteomic analysis. The results showed that CAP significantly reduced the expression of HSP90AB1 and SMARCC1 in the spleen. It was subsequently confirmed that CAP also significantly reduced the expression of HSP90AB1 and SMARCC1 and increased ROS and apoptosis levels in T cells. The results of in vitro experiments showed that HSP90AB1 knockdown resulted in a significant decrease in p-Akt, SMARCC1, p-c-Fos, and p-c-Jun expression levels and a significant increase in ROS and apoptosis levels. HSP90AB1 overexpression significantly inhibited CAP-induced T cell apoptosis by increasing the p-Akt, SMARCC1, p-c-Fos, and p-c-Jun expression levels and reducing the ROS level. In conclusion, HSP90AB1 is a key target of CAP-induced T cell apoptosis via Akt/SMARCC1/AP-1/ROS axis, which provides a novel understanding of CAP-induced T cell apoptosis and lays the experimental foundation for further exploring CAP as an immunosuppressant with antitumor effects to optimize the medication regimen for transplantation patients.

This paper investigates an effective controller design method for discrete-time linear system with parameter uncertainty, time-varying delay, input nonlinearity and process disturbance. A novel dis...

Low-cost particulate matter (PM) sensors can be widely deployed to measure aerosol concentrations at higher spatial and temporal resolutions than traditional instruments, but they need to be carefully calibrated under ambient conditions. In this study, a long-term field experiment was conducted from December 2015 to May 2017 at a site in Nanjing to evaluate the capabilities of in-house built low-cost PM monitors using the Shinyei PPD42NS sensor for ambient PM2.5 monitoring. A BAM-1020 particulate monitor was co-located with the low-cost sensors to provide reference readings. Least-square regressions with linear and power-law functions, and an artificial neural network (ANN) technique were used to convert electrical instrument readings to ambient aerosol concentrations. Applying the ANN technique resulted in the best estimation of the hourly PM2.5 (R2 = 0.84; mean normalized bias = 12.7% and mean normalized error (MNE) = 29.7%). The low-cost sensors displayed relatively good performance with high aerosol concentrations but larger errors with concentrations below 35 µg m–3. High humidity (RH > 75%) can cause a larger MNE for these sensors, but the impact of temperature was negligible in this study. A clear sensor deterioration trend was observed during the 18-month field calibration. High correlations were found between the data from a single low-cost sensor and the data from the BAM-1020 when the low-cost sensor was individually calibrated, but the correlations between measurements taken by different low-cost sensor units were only moderate, possibly due to internal sensor variations. The results suggest that these low-cost sensors can measure ambient PM2.5 concentrations with an acceptable level of accuracy, which can and should be improved by calibrating each sensor individually. Special attention should be paid to the accuracy of these sensors after long-term application and in highly humid environments.

The experimental evidences linking PM

Fine particulate matter (PM2.5 ) concentrations show high variations in different microenvironments indoors, which has considerable impact on risk management. However, the real-time variations of PM2.5 exposure associated with per activity/microenvironment and intra-variation among family members remain undefined. In this study, real-time monitors were used to collect real-time PM2.5 data in different microenvironments in 32 households in urban community of China. Peak concentrations of PM2.5 were found in kitchen. The parallel levels of PM2.5 household indoor and outdoor indicated the benefit of clean energies use. To validly assess the health risk of individuals, we proposed a novel method to estimate the real-time exposure of all residents and firstly investigate the intra-variation of PM2.5 exposure among family members. The member who is responsible for cooking in the family had the maximum PM2.5 exposure. The ratios among intraindividual variations demonstrated children usually had lower exposure compared to the adults as they stayed more time in lower polluted microenvironments such as living room and bedroom. The exposure intensity in living room was above 1.0 for most residents, indicating it is warranted to alleviate the air pollution in living room. This study firstly focused on the intra differences of PM2.5 exposure among family members and provided a new insight for indoor air pollution management. The results suggested when adopting measures to reduce exposure, the microenvironments pattern of each member should be taken into consideration. Future work is welcomed to move another big step on this issue to protect the human health.

Fine particulate matter (PM

The selection and spread of antibiotic resistance poses risks to public health by reducing the therapeutic potential of antibiotics against human pathogens. Wastewater-based epidemiology (WBE) is potentially the most reliable approach to estimate antibiotics use. Previous WBE studies used parent antibiotics as biomarkers, which may lead to overestimation since parent antibiotics may be directly disposed of. Using metabolites as biomarkers can avoid this drawback. This study developed a simultaneous solid-phase extraction coupled with ultra-high-performance liquid chromatography tandem mass spectrometry method for analyzing 12 antibiotics and human metabolites in wastewater to help assess health risk. Optimum conditions were achieved using a PEP cartridge at pH 3.0. The extraction efficiencies were 73.3~95.4% in influent and 72.0~102.7% in effluent for most of the target analytes. Method detection limit ranged from 0.1 to 1.5 ng/L for influent wastewater and 0.03 to 0.7 ng/L for effluent wastewater. A stability experiment showed that sulfonamide parents and their metabolites were stable at 4 °C, −20 °C and −80 °C, while macrolides metabolites were more stable than their corresponding parents at 4 °C and −20 °C. Finally, the method was applied to measure these analytes in wastewater samples collected from three Beijing WWTPs and to derive apparent removal rates. All metabolites were detected in wastewater samples with concentrations ranging from 1.2 to 772.2 ng/L in influent, from

Background The advanced hepatocellular carcinoma (HCC), such as the recurrent tumor after liver transplantation (LT), is an obstacle of HCC treatment. The aim of this study was to discover the underlying mechanism of HCC progression caused by non-coding RNAs (ncRNAs). Methods To this end, we investigated the selected patient cohort of matching primary and recurrent HCC after receiving LT. The recurrent tumors after LT were regarded as clinical models of the advanced HCC. Microarrays were used to profile lncRNA and mRNA expression in HCC recurrent and primary tissue samples. The mRNA profile characteristics were analyzed by bioinformatics. Two cell lines, HepG2 and QGY-7703, were used as HCC cell models. The protein-coding potential, length, and subcellular location of the interested lncRNAs were examined by bioinformatics, Northern blot, fluorescent in situ hybridization (FISH), and quantitative RT-PCR (qRT-PCR) assays. HCC cell proliferation was detected by CCK-8, doubling time and proliferation marker gene quantitation assays. DNA replication during the cell cycle was measured by EdU/PI staining and flow cytometry analyses. Promoter activity was measured using a luciferase reporter assay. Interactions between DNA, RNA, and protein were examined by immunoprecipitation and pull-down assays. The miRNA-target regulation was validated by a fluorescent reporter assay. Results Both lncRNA and mRNA profiles exhibited characteristic alterations in the recurrent tumor cells compared with the primary HCC. The mRNA profile in the HCC recurrent tissues, which served as model of advanced HCC, showed an aberrant cell cycle regulation. Two lncRNAs, the highly expressed lncRNA in recurrent HCC (HERH)-1 and HERH-4, were upregulated in the advanced HCC cells. HERH-1/4 enhanced proliferation and promoted DNA replication and G1-S transition during the cell cycle in HCC cells. HERH-1 interacted with the transcription factor CREB1. CREB1 enhanced cyclin A2 (CCNA2) transcription, depending on HERH-1-CREB1 interaction. HERH-4 acted as an miR-29b/c sponge to facilitate CCNA2 protein translation through a competing endogenous RNA (ceRNA) pathway. Conclusions The oncogenic lncRNA HERH-1/4 promoted CCNA2 expression at the transcriptional and post-transcriptional levels and accelerated cell cycle progression in HCC cells. The HERH-1-CREB1-CCNA2 and HERH-4-miR-29b/c-CCNA2 axes served as molecular stimuli for HCC advance.

To explore gestational diabetes mellitus (GDM) diagnostic markers and establish the predictive model of GDM.We downloaded the DNA methylation data of GSE70453 and GSE102177 from the Gene Expression Omnibus database. Epigenome-wide association study (EWAS) was performed to analyze the relationship between cytosine-phosphate-guanine (CpG) methylation and GDM. And then the logistic regression models were constructed, with the β-values of CpG sites as predictor variable and the GDM occurrence as binary outcome variable. Data from GSE70453 served as training sets and data from GSE102177 served as verification sets.The EWAS and overlap analysis identified nine-shared significant CpGs in the two DNA methylation data sets. Remarkably, these nine CpGs were differently methylated in GDM samples compared to their matched normal specimens, among which five fully methylated CpGs were finally selected. Importantly, we established a binary logistic regression model based on the above five CpGs, in which cg11169102, cg21179618 and cg21620107 were critical. Hence, we further built a logistic regression model by using the three CpGs and found that the area under the curve was 0.8209. The validation of the model by using the verification sets indicated the area under the curve was 0.8519.We identified potential CpG biomarkers for the diagnosis of gestational diabetes mellitus patients through using EWAS and Logistic regression models in combination.

Pulmonary hypertension (PH), a rare but deadly cardiopulmonary disorder, is characterized by extensive remodeling of pulmonary arteries resulting from enhancement of pulmonary artery smooth muscle cell proliferation and suppressed apoptosis; however, the underlying pathophysiological mechanisms remain largely unknown. Recently, epigenetics has gained increasing prominence in the development of PH. We aimed to investigate the role of vestigial-like family member 4 (VGLL4) in chronic normobaric hypoxia (CNH)-induced PH and to address whether it is associated with epigenetic regulation. The rodent model of PH was established by CNH treatment (10% O2 , 23 hours/day). Western blot, quantitative reverse transcription polymerase chain reaction, immunofluorescence, immunoprecipitation, and adeno-associated virus tests were performed to explore the potential mechanisms involved in CNH-induced PH in mice. VGLL4 expression was upregulated and correlated with CNH in PH mouse lung tissues in a time-dependent manner. VGLL4 colocalized with α-smooth muscle actin in cultured pulmonary arterial smooth muscle cells (PASMCs), and VGLL4 immunoactivity was increased in PASMCs following hypoxia exposure in vitro. VGLL4 knockdown attenuated CNH-induced PH and pulmonary artery remodeling by blunting signal transducer and activator of transcription 3 (STAT3) signaling; conversely, VGLL4 overexpression exacerbated the development of PH. CNH enhanced the acetylation of VGLL4 and increased the interaction of ac-H3K9/VGLL4 and ac-H3K9/STAT3 in the lung tissues, and levels of ac-H3K9, p-STAT3/STAT3, and proliferation-associated protein levels were markedly up-regulated, whereas apoptosis-related protein levels were significantly downregulated, in the lung tissues of mice with CNH-induced PH. Notably, abrogation of VGLL4 acetylation reversed CNH-induced PH and pulmonary artery remodeling and suppressed STAT3 signaling. Finally, STAT3 knockdown alleviated CNH-induced PH. In conclusion, VGLL4 acetylation upregulation could contribute to CNH-induced PH and pulmonary artery remodeling via STAT3 signaling, and abrogation of VGLL4 acetylation reversed CNH-induced PH. Pharmacological or genetic deletion of VGLL4 might be a potential target for therapeutic interventions in CNH-induced PH.

BackgroundNeoantigen (NeoAg) peptides displayed at the tumor cell surface by human leukocyte antigen molecules show exquisite tumor specificity and can elicit T cell mediated tumor rejection. However, few NeoAgs are predicted to be shared between patients, and none to date have demonstrated therapeutic value in the context of vaccination.MethodsWe report here a phase I trial of personalized NeoAg peptide vaccination (PPV) of 24 stage III/IV non-small cell lung cancer (NSCLC) patients who had previously progressed following multiple conventional therapies, including surgery, radiation, chemotherapy, and tyrosine kinase inhibitors (TKIs). Primary endpoints of the trial evaluated feasibility, tolerability, and safety of the personalized vaccination approach, and secondary trial endpoints assessed tumor-specific immune reactivity and clinical responses. Of the 16 patients with epidermal growth factor receptor (EGFR) mutations, nine continued TKI therapy concurrent with PPV and seven patients received PPV alone.ResultsOut of 29 patients enrolled in the trial, 24 were immunized with personalized NeoAg peptides. Aside from transient rash, fatigue and/or fever observed in three patients, no other treatment-related adverse events were observed. Median progression-free survival and overall survival of the 24 vaccinated patients were 6.0 and 8.9 months, respectively. Within 3–4 months following initiation of PPV, seven RECIST-based objective clinical responses including one complete response were observed. Notably, all seven clinical responders had EGFR-mutated tumors, including four patients that had continued TKI therapy concurrently with PPV. Immune monitoring showed that five of the seven responding patients demonstrated vaccine-induced T cell responses against EGFR NeoAg peptides. Furthermore, two highly shared EGFR mutations (L858R and T790M) were shown to be immunogenic in four of the responding patients, all of whom demonstrated increases in peripheral blood neoantigen-specific CD8+ T cell frequencies during the course of PPV.ConclusionsThese results show that personalized NeoAg vaccination is feasible and safe for advanced-stage NSCLC patients. The clinical and immune responses observed following PPV suggest that EGFR mutations constitute shared, immunogenic neoantigens with promising immunotherapeutic potential for large subsets of NSCLC patients. Furthermore, PPV with concurrent EGFR inhibitor therapy was well tolerated and may have contributed to the induction of PPV-induced T cell responses.

Background Highly upregulated in liver cancer (HULC) is a long non-coding RNA (lncRNA) which has recently been identified as a key regulator in hepatocellular carcinoma (HCC) progression. However, its role in the secretion of exosomes from HCC cells remains unknown. Aim To explore the mechanism by which HULC promotes the secretion of exosomes from HCC cells. Methods Serum and liver tissue samples were collected from 30 patients with HCC who had not received chemotherapy, radiotherapy, or immunotherapy before surgery. HULC expression in serum exosomes and liver cancer tissues of patients was measured, and compared with the data obtained from healthy controls and tumor adjacent tissues. The effect of HULC upregulation in HCC cell lines and the relationship between HULC and other RNAs were studied using qPCR and dual-luciferase reporter assays. Nanoparticle tracking analysis was performed to detect the quantity of exosomes. Results HULC expression in serum exosomes of patients with HCC was higher than that in serum exosomes of healthy controls, and HULC levels were higher in liver cancer tissues than in tumor adjacent tissues. The expression of HULC in serum exosomes and liver cancer tissues correlated with the tumor-node-metastasis (TNM) classification, and HULC expression in tissues correlated with that in serum exosomes. Upregulation of HULC promoted HCC cell growth and invasion and repressed apoptosis. Notably, it also facilitated the secretion of exosomes from HCC cells. Moreover, qPCR assays showed that HULC repressed microRNA-372-3p (miR-372-3p) expression. We also identified Rab11a as a downstream target of miR-372-3p. Dual-luciferase reporter assays suggested that miR-372-3p could directly bind both HULC and Rab11a. Conclusion Our findings illustrate the importance of the HULC/miR-372-3p/Rab11a axis in HCC and provide new insights into the molecular mechanism regulating the secretion of exosomes from HCC cells.

Toll‐like receptor 4 (TLR4) plays an essential role in cancer progress. Here, we find that the expression of TLR4 in relapsed human hepatocellular carcinoma (HCC) clinical samples is higher than that in the non‐relapsed ones, which leads us to explore the role of TLR4 in cancer stemness. We reported that TLR4‐AKT signaling pathway was activated by lipopolysaccharides (LPS) in HCC cell lines to enhance the cancer stemness capacity, which was reflected by the increased percentage of CD133+ CD49f+ population and side population, enhanced sphere formation, and the upregulation of stemness marker gene‐SOX2. Downregulation of SOX2 attenuated the enhanced HCC stemness induced by LPS, indicating SOX2 as a downstream mediator of LPS‐TLR4 signaling. The role of LPS‐TLR4 signaling in inducing HCC stemness was further confirmed by tumor xenograft experiment in vivo. Taken together, our findings provide a novel therapeutic target to prevent the recurrence of HCC.

Hepatocellular carcinoma (HCC) is the third most common cause of cancer mortality worldwide, which is partially due to the lack of appropriate therapeutic options. The development of HCC is accompanied with unique and continuous genomic and epigenetic modifications. Therefore, the absence of a personalized and reproducible human model reduces the ability to determine the potential of candidate treatments. Conditional reprogramming (CR) culture has been used to establish and indefinitely grow patient‑derived tumor cell lines in a rapid and efficient manner. In the present study, primary HCC cells were isolated from tumor specimens and cultured under CR conditions. The proliferative potential and capacity of cells to undergo continuous regeneration were evaluated by cell viability and proliferation assays, and the expression of tumor‑specific markers was determined by western blotting and immunofluorescence to determine the prospects for use in clinical settings. It was demonstrated that ~55% of tumor samples were able to generate HCC cells that could be continuously expanded and passaged under CR conditions; this ability was associated with the source and composition of the tumor tissues. Furthermore, the expression of the tumor‑specific marker α‑fetoprotein and the proliferative ability of cells were maintained following cycles of cryopreservation and resuscitation. In conclusion, with further optimization, the CR system may be a useful tool for the precise therapeutic treatment of patients with HCC.

The temporal variation of antibiotics and ARGs as well as the impact of daily schedule of inpatients on their regular occurrence in hospital wastewater (HWW) were previously obscure. In this study, the wastewater of the inpatient department pre- and posttreatment (hydraulic retention time = 8 h) was collected intraday and intraweek. The absolute concentrations of antibiotics/metabolites and ARGs in HWW were analyzed to investigate the temporal variations of their occurrence levels. Fluoroquinolones were the predominant drugs used in the inpatient department (681.30-881.66 ng/mL in the effluent) and the main contaminant in the outlet of the disinfection pond (538.29-671.47 ng/mL). Diurnal variations peaked at 19:00 for most antibiotics and ARGs, while the maximum of them occurred on weekends. Aminoglycoside resistance genes (AMRGs, 21.6-23000 copies/mL) and β-lactam resistance genes (BLGRs, 1.24-8500 copies/mL) were the dominant ARGs before and after treatment processing, respectively (p 0.05). The significant removal rates (50%) of most antibiotics and ARGs, as well as the integrase gene intI1 and 16S rRNA gene, were found to be subjected solely to the chloride disinfection process, suggesting the necessity of the self-contained wastewater treatment process. Meanwhile, the statistically significant correlation among antibiotics, ARGs, intI1, and 16S rRNA (p 0.05) demonstrated that the risk of selective pressure, horizontal transfer and vertical propagation of ARGs in the effluent of the hospital was warranted. Principal component analysis (PCA) showed that the daily schedule of inpatients and wastewater treatment processes could markedly induce fluctuations in antibiotic and ARG levels in HWW, indicating that they should be considered an impact factor for environmental monitoring. This study demonstrated for the first time the temporal variations in the abundance and dissemination of antibiotics and ARGs in a semiclosed zone and provided new insight into the development of assessments of the associated ecological risk and human health.

Postoperative hepatocellular carcinoma (HCC) recurrence and metastasis throw great threaten to its overall survival (OS). This paper focus on exploring the prognostic significance of NANOG and OCT4 expression in HCC recurrence and OS after liver transplantation. Eighty-six patients who meet University of California San Francisco (UCSF) criteria and underwent liver transplantation in Tianjin First Central Hospital between August 2010 and August 2013 were included. Expression of NANOG and OCT4 was determined by immunohistochemistry. The relationships between NANOG and OCT4 expression with tumor recurrence, tumor count, histology stage, lymph node metastasis (LNM) and microvascular invasion (MVI) were explored through the χ2 test and Cox regression analysis. We found that 19/26 and 20/24 patients with positive expression of NANOG and OCT4 relapsed. Combination of NANOG and OCT4 expression was indicated as valuable prognostic signature for HCC recurrence prediction (P < 0.0011). Besides, we identified other key factors with significant correlations with recurrence, such as LNM (P = 0.011) and MVI (P = 0.024). Strikingly, recurrence sites could significantly affect recurrence time (P = 0.0062) and patients with recurrence in transplanted liver have longer recurrence time. In conclusions, we analyzed the relationships between NANOG/OCT4 expression, clinicopathology features, HCC recurrence, and OS after liver transplantation for the first time. Combination of NANOG, OCT4, LNM, histopathological stage, and MVI may be predictor for HCC recurrence posttransplantation. Comprehensive of histopathological stage grade and LNM were considered as prognosis factor for OS after liver transplantation. This should be helpful for treatment method selection for HCC patients after liver transplantation.

Methamphetamine (METH) and ketamine (KET) are widely detected in surface waters and thus may pose threat to aquatic organisms. However, their degradation in aquatic systems and the effects on bacterial community were unknown. The present study investigated the biodegradation process of METH and KET in river waters and sediments. Three microcosms were examined over 40-days’ incubation under (i) aerobic and illumination conditions, (ii) anaerobic condition exposed to light, (iii) anaerobic-dark condition. Statistically significant biodegradation of METH and KET (1 mg L−1) was observed in all treatments. The half-lives under the examined conditions indicate that the two drugs were refractory in aquatic environment. Moreover, there were no pronounced absorption and photolysis observed in this work. Illumina MiSeq sequencing analysis revealed that Methylophilaceae, Saprospiraceae, WCHB1–69, Desulfobulbaceae, Porphyromonadaceae, FamilyXI, Peptococcaceae, and Rhizobiaceae were the predominant candidatus families during KET and METH biodegradation, and the preponderance would impair other microorganisms’ prosperity since them were scarcely detected in the wild. Meanwhile, canonical correlation analysis (CCA) indicates that METH as an environmental factor may affect bacterial community structure in field water samples.

Evidence about the adverse effects of methamphetamine (METH) on invertebrates is scarce. Hence, C. elegans, a representative invertebrate model, was exposed to METH at environmental levels to estimate chronic and transgenerational toxicity. The results of chronic exposure were integrated into an underlying toxicity framework of METH in invertebrates (e.g., benthos) at environmentally relevant concentrations. The induction of cellular oxidative damage-induced apoptosis and fluctuation of ecologically important traits (i.e., feeding and locomotion) might be attributed by the activation of the longevity regulating pathway regulated by DAF-16/FOXO, and detoxification by CYP family enzymes. The adverse effects to the organism level included impaired viability and decreased fecundity. The results from transgenerational exposure elucidated the cumulative METH-induced damage in invertebrates. Finally, a new risk assessment method named toxicity indicator sensitivity distribution (TISD) analysis was proposed by combining multiple toxicity indicator test data (ECx) to derive the hazardous concentration for 10% indicators (C10) of one species. The risk quotient (RQ) values calculated by measured environmental concentrations and C10 in southern China, southeastern Australia, and the western US crossed the alarm line (RQ = 5), suggesting a need for long-term monitoring.

The adverse effects on model fish induced by methamphetamine (METH) have been revealed. However, the toxicity of METH on different kinds of non-model fish during the natural attenuation remained unclear. Hence, in this study, we for the first time established a static lab-scale aquatic ecosystem spiked with METH (initial levels at 25 μg/L) for 40 days to estimate its metabolism and toxicity in Chinese medaka, rosy bitterling, loach, and mosquito fish. The concentrations of METH in water and fish's brain were detected termly. The physiological functions, histopathology of brain, neurotransmitters contents, and expressions of associated genes of the four kinds of fish were determined at day 0, 20, and 40, respectively. The results indicated METH could be remarkably accumulated in fish brains with the distribution factor vs water (DFw) at 232.5-folds, and attenuated both in water and fish body during the exposure. METH caused physiological functions (i.e., swimming trajectories, locomotion distances, and feeding rates) disorders of the four kinds of fish, and stimulated surfacing behavior of loach. Tissue and macro/micromolecular biomarkers including histopathology, neurotransmitters (i.e., dopamine, serotonin, and norepinephrine), and mRNA, were similarly affected by METH. Mitogen-activated protein kinase (MAPKs) signaling pathway, P53-regulated apoptosis signaling pathway, N-methyl- d -aspartate-dopamine system, and mTOR signaling pathway of different kinds of fish were regulated by METH. Additionally, the impairments of the physiological and macromolecular indicators of fish could be alleviated as the natural attenuation of METH occurred. All the biomarkers, as well as the recovery effects during the exposure were integrated onto an adverse outcome pathway (AOP) framework. The key event was the micromolecular indicators (genes). The adverse outcomes at individual and population levels would result in the ecological consequences, implying the imperative to consider the natural attenuation process while assessing the environmental risk of METH.