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Traumatic brain injury refers to brain injury caused by mechanical impact often leading to severe morbidity and mortality. Despite increasing awareness, there are no effective treatments strategies. Therefore, there is a need to develop new effective treatments for this injury. Forsythiaside A is a monomer of phenylethanolglucoside extracted from Forsythia, which has a wide range of pharmacological properties including protective effects on brain tissue. Herein, using a rat model of traumatic brain injury, we have shown that forsythiaside A can improve nerve function and brain tissue injury in rats with traumatic brain injury, and reduce brain inflammation and neuronal apoptosis. We have further shown that forsythiaside A regulates toll-like receptor 4/myeloid differentiation factor 88/nuclear factor-kappa B signaling pathway. This opens the possibility of a potentially promising therapeutic drug for the treatment of traumatic brain injury.

Summary Epithelial regeneration is critical for barrier maintenance and organ function after intestinal injury, although the repair mechanisms are unclear. Here, we found that Bach2 deficiency promotes intestinal epithelial cell proliferation during homeostasis. Moreover, genetic inactivation of Bach2 in mouse intestinal epithelium facilitated crypt regeneration after irradiation, resulting in a reduction in mortality. RNA-sequencing analysis of isolated crypts revealed that Bach2 deficiency altered the expression of numerous genes, including those regulating double-strand break repair. Mechanistic characterizations indicated that Bach2 deletion facilitated DNA repair in intestinal crypt cells, as evidenced by faster resolution of γ-H2AX and 53BP1 foci in Bach2−/− crypt cells, compared with Bach2+/+ control. Together, our studies highlight that Bach2 deficiency promotes intestinal regeneration by accelerating DNA repair in intestinal stem cells after radiation damage., Graphical Abstract, Highlights • Bach2 deficiency facilitates intestinal recovery after irradiation damage • Bach2 deficiency promotes the regeneration of crypt intestinal stem cells (ISCs) • Bach2 deletion accelerates DNA repair in ISCs, Radiation causes intestinal injury, as occurs in abdominal and pelvic tumor radiotherapy. Intestinal stem cells (ISCs) are critical for epithelial regeneration. In this article, Hua and colleagues show that Bach2 deficiency promotes intestinal epithelial regeneration by accelerating DNA repair in ISCs, resulting in a reduction in mortality during radiation-induced intestine injury.

Cleaner winter heating has been promoted to abate the winter air pollution in northern China. Although improvements in air quality have been observed, the effectiveness and mechanism of cleaner heating measures on air quality have not been examined on the empirical ground. In this study, we estimate the annual effects of winter heating policy on air quality from 2014 to 2017 using a regression discontinuity design (RDD) and dynamic regression model. The results show that winter heating aggravates Air Quality Index (AQI). Specifically, the AQI raised by winter heating reduce from 85.3 in 2014 to 24.1 in 2017, indicating diminishing effects of winter heating with the implementation of clean heating measures. The heterogeneous characteristics of winter heating in terms of different pollutants and city scales are further quantified. The effects of clean heating are more evident for particulate pollutants (PM

Background Gallbladder carcinoma (GBC) is one of the most lethal malignancies which do not have a targeted drug in the clinic. Patient-derived primary cell lines (PDCs) are useful in assessment of cancer complexity and heterogeneity, drug-sensitivity tests, and personalized-drug-selection guidance. The aim of this study is to establish GBC PDCs and characterize their biological features. Methods The characterization of PDCs was defined by morphology, growth kinetics, chromosomal analysis, short tandem repeat (STR) analysis, RNA-seq and tumorigenicity. Glycosylation of PDCs derived from GBC was first studied, and the PDC model’s performance were also tested and evaluated using seven molecular target inhibitors. Results Three novel GBC cell lines from three GBC patients were successfully established and denoted as JXQ-3D-902R4, JXQ-3D-4494R, and JXQ-3D-4786R. These cell lines demonstrated the heterogeneous characteristics of tumor morphology and phenotypes which are consistent with primary GBC, such as irregular cell shape, varied chromosomal numbers, and different STR patterns. Moreover, the growth activity and tumorigenicity ability varied among the cell lines, of which JXQ-3D-4494R exhibited the best growth rate. Furthermore, glycan profiling of whole proteins were detected and characterized. Unique N-glycans of each PDC were identified, JXQ-3D-902R4, JXQ-3D-4494R and JXQ-3D-4786R contained ten, four and seven unique glycans, respectively. The epithelial origins of three PDCs were confirmed using RNA-seq based on the highly expressed typical epithelial marker genes. Moreover, the drug-sensitivity results demonstrated that the three PDCs exhibited different responses to the seven-most commonly used targeted medicines belonging to three groups: cell-cycle inhibitors, PI3K/AKT/mTOR signaling-pathway inhibitors, and ErbB inhibitors. JXQ-3D-4494R was sensitive to most of the inhibitors, JXQ-3D-4786R was sensitive to ErbB inhibitors, and JXQ-3D-902R4 was sensitive to PI3K/AKT/mTOR inhibitors. Conclusions These results indicate that PDCs may be efficient preclinical models for further investigation of the biological behaviors and potential targeted therapies of human GBC.

Methane oxidation coupled to denitrification is mediated by Candidatus "Methylomirabilis oxyfera" (M. oxyfera), which belongs to the candidate phylum NC10, and plays a crucial role in the global carbon and nitrogen cycle. Using the Yellow River Delta coastal wetland as the study area, molecular biology technology and laboratory incubation were used to determine the abundance of NC10 bacteria and the denitrifying anaerobic methane oxidation (DAMO) rate in soils from different vegetation areas. The results of the electrophoresis detection show that M. oxyfera-like bacteria can be found in the four types of soils, according to the growth analysis by the system, OTU1 (SA) has been found the highest similarity to first-discovered Candidatus Methylomir-abilis oxyfera (FP565575) (over 98%); Vegetation cover significantly increased the abundance of M. oxyfera-like bacteria compared to beach areas, which abundance was significantly higher in deeper layers than in surface ones. Nitrate, nitrite, total nitrogen, and conductivity were identified as the main environmental factors affecting the DAMO rate. This study showed that both groups A and B of Candidatus M. oxyfera-like bacteria exist in the coastal wetland of the Yellow River Delta, which provides molecular biological evidence for the existence of the DAMO process therein. Moreover, it was revealed the influence mechanism of physical and chemical characteristics of each region on the DAMO rate. This is of significance for furthering our understanding of the coupled effect of the global carbon and nitrogen cycle.

Benign recurrent meningitis, also known as Mollaret’s meningitis (MM), is characterized by recurrent headache, fever, meningeal irritation and sterility of cerebrospinal fluid examination, which is rare in clinic. Although with clear diagnostic criteria, there are still many unclear aspects of its pathogenesis, and treatment of this disease. The author reported a case of benign recurrent aseptic meningitis, who had three episodes of acute headache with or without fever. The patient was treated with ceftriaxone, acyclovir and cefazoxime for the first time, cefazoxime, cefimidazole and mebendazole for the second time, and piperacillin and acyclovir for the third time. The results showed that the patient did not recover completely in the first two times and fully recovered at the third time. Based on the current research, the author believes that for MM, mainly symptomatic relief, indomethacin should be recommended, and too much antiviral treatment is not needed.

The bismuth vanadate (BiVO4) photoanode receives extensive attention in photoelectrochemical (PEC) water splitting. However, the high charge recombination rate, low electronic conductivity, and sluggish electrode kinetics have inhibited the PEC performance. Increasing the reaction temperature for water oxidation is an effective way to enhance the carrier kinetics of BiVO4. Herein, a polypyrrole (PPy) layer was coated on the BiVO4 film. The PPy layer could harvest the near-infrared light to elevate the temperature of the BiVO4 photoelectrode and further improve charge separation and injection efficiencies. In addition, the conductive polymer PPy layer acted as an effective charge transfer channel to facilitate photogenerated holes moving from BiVO4 to the electrode/electrolyte interface. Therefore, PPy modification led to a significantly improved water oxidation property. After loading the cobalt–phosphate co-catalyst, the photocurrent density reached 3.64 mA cm−2 at 1.23 V vs the reversible hydrogen electrode, corresponding to an incident photon-to-current conversion efficiency of 63% at 430 nm. This work provided an effective strategy for designing a photothermal material assisted photoelectrode for efficient water splitting.

SIMILAR TO RCD ONE (SRO) gene family is a small plant-specific gene family responsible for growth, development, and stress responses. In particular, it plays a vital role in responding to abiotic stresses such as salt, drought, and heavy metals. Poplar SROs are rarely reported to date. In this study, a total of nine SRO genes were identified from Populus simonii × Populus nigra, which are more similar to dicotyledon SRO members. According to phylogenetic analysis, the nine PtSROs can be divided into two groups, and the members in the same cluster have a similar structure. There were some cis-regulatory elements related to abiotic stress response and hormone-induced factors identified in the promoter regions of PtSROs members. Subcellular localization and transcriptional activation activity of PtSRO members revealed a consistent expression profile of the genes with similar structural profiles. In addition, both RT-qPCR and RNA-Seq results indicated that PtSRO members responded to PEG-6000, NaCl, and ABA stress in the roots and leaves of Populus simonii × Populus nigra. The PtSRO genes displayed different expression patterns and peaked at different time points in the two tissues, which was more significant in the leaves. Among them, PtSRO1c and PtSRO2c were more prominent in response to abiotic stress. Furthermore, protein interaction prediction showed that the nine PtSROs might interact with a broad range of transcription factors (TFs) involved in stress responses. In conclusion, the study provides a solid basis for functional analysis of the SRO gene family in abiotic stress responses in poplar.

Anaerobic oxidation of methane (AOM) in wetland soils is widely recognized as a key sink for the greenhouse gas methane (CH

Significant technological advancements in the Internet of Vehicles (IoV) incentivize the rapid development of various vehicular communication technologies for wireless connectivity among vehicles, roadside devices, and pedestrians. Despite the countless benefits, security issues have received considerable critical attention since breaches have been recurrent in vehicular communication networks and applications, which seriously affects the transportation environment. To resolve the security issues in vehicular communication, we introduce a novel signature scheme based on a standard elliptic curve cryptography (ECC) algorithm in this paper. Compared with existing schemes, the proposed scheme is lightweight and cost-saving in the vehicular environment and can efficiently authenticate and protect the exchange of messages between each entity in the IoV. The security of our scheme is based on the unsolved NP-complete problem: the elliptic curve discrete logarithm problem. Moreover, by using key prefixing, we demonstrate the merits of our scheme through security analysis in terms of avoiding key escrow and replacement attack resistance.

Estrogen is very important to the differentiation of B lymphocytes; B lymphopoiesis induced by OVX was supposedly involved in osteoporosis. But the effects of B lymphocytes on the osteogenic differentiation of bone mesenchymal stem cells (BMSCs) are not clear. In this study, we detected bone quality and bone loss in a trabecular bone by electronic universal material testing machine and microcomputed tomography (micro-CT) in OVX and splenectomized-ovariectomy (SPX-OVX) rats. Additionally, changes in lymphocytes (B lymphocyte, CD4+ and CD8+ T lymphocytes, and macrophages) in the bone marrow were analyzed by flow cytometry. The osteogenesis of BMSCs cocultured with normal and LPS-pretreated B lymphocytes was detected by BCIP/NBT and Alizarin red S staining. Measurement of the Notch2, Notch4, Hey1, Hey2, Hes1, and runt-related transcription factor 2 (Runx2) expression in BMSCs cocultured with B lymphocytes was done using real-time PCR. The effects of dexamethasone and DAPT (inhibitor of Notch signaling) on osteogenesis of BMSCs were detected by BCIP/NBT, Alizarin red S staining, and real-time PCR. Osteoporosis happened in OVX rats, more serious in SPX-OVX rats, B lymphocytes increased in OVX rats, and sharply higher in SPX-OVX rats. Osteoporosis did not happen in SPX rats which is still companied with a high increase of B lymphocytes. LPS-pretreated B lymphocytes suppressed the osteogenesis of BMSCs, but the normal B lymphocytes could not. The LPS-pretreated B lymphocytes upregulated the expression of Notch4, Hes1, and Hey2 and downregulated the expression of Runx2 in BMSCs. Dexamethasone and DAPT could downregulate the high expression of Notch4, Hes1, Hey2 and upregulate the low expression of Runx2 in BMSCs which cocultured with LPS treated B lymphocytes, the inhibited ALP and Alizarin red staining in BMSCs which cocultured with LPS treated B lymphocytes also partly restored.

Chronic and systemic bone complications frequently occur in patients who undergo radiotherapy; however, the pathological mechanisms underlying these complications remain unclear. This study aimed to observe persistent and systemic changes in locally irradiated rats and to determine the systemic pathological changes that persistently affect bone metabolism. We examined the inflammatory and oxidative stress responses that occurred after local irradiation using enzyme immunoassays and biochemical analyses. Lymphocytes obtained from the blood, spleen, thymus, and bone marrow were evaluated using flow cytometry. The proliferation and apoptosis characteristics of co-cultured bone marrow-derived mesenchymal stem cells (BMSCs) were detected by MTT assay and PI/Annexin V-FITC staining, respectively, and the differentiation of BMSCs was measured according to alkaline phosphatase (ALP) staining, alizarin red staining, and Oil Red O staining and by evaluating the mRNA expression of ALP, osteocalcin (OCN), osteopontin (OPN), collagen I, Runx2, and PPARγ. Our results revealed that no significant or continuous differences were present in the inflammatory response or the oxidative stress response throughout the body after local irradiation. B lymphocyte levels increased continuously in the blood, spleen, and bone marrow after local irradiation. T lymphocyte levels were decreased at 2 weeks after local irradiation, and CD8+T lymphocyte levels were increased in the blood, thymus, and bone marrow at 12 weeks after local irradiation. The ratio of CD4+/CD8+T lymphocytes began to decrease during the early phase after local irradiation and became significantly decreased at 12 weeks after local irradiation. Normal BMSCs co-cultured with lymphocytes derived from irradiated rats exhibited decreased proliferation and increased apoptosis, and the ALP staining intensity, alizarin red staining intensity, and mRNA expression of related genes were all also decreased. Oil Red O staining intensity and mRNA expression of PPARγ were both increased. Lymphocyte levels contribute to chronic and systemic bone complications after radiotherapy by inhibiting the proliferation and osteoblastogenesis of BMSCs.

In response to the spread of COVID-19, China implemented a series of control measures. The causal effect of these control measures on air quality is an important consideration for extreme air pollution control in China. Here, we established a difference-in-differences model to quantitatively estimate the lockdown effect on air quality in the Beijing-Tianjin-Hebei (BTH) region. We found that the lockdown measures did have an obvious effect on air quality. The air quality index (AQI) was reduced by 15.2%, the concentration of NO2, PM10, PM2.5, and CO were reduced by 37.8%, 33.6%, 21.5%, and 20.4% respectively. At the same time, we further explored the heterogeneous effects of travel restrictions and the control measure intensity on air quality. We found that the traffic restrictions, especially the restriction of intra-city travel intensity (TI), exhibited a significant heterogeneous effect on NO2 with a decrease of approximately 13.6%, and every one-unit increase in control measures intensity reduced the concentration of air pollutants by approximately 2–4%. This study not only provides a natural, experimental basis for control measures on air quality but also indicates an important direction for future control strategies. Importantly, determining the estimated effect helps formulate accurate and effective intervention measures on the differentiated level of air pollution, especially on extreme air pollution.

e15598 Background: 5-FU-based regimens are the mainstay treatment for colorectal cancer. However, the limited response rate to 5-FU-based regimens hampered the increase in the treatment efficacy of locally advanced rectal cancer. During the drug screening in the rectal cancer organoid biobank (n = 106), we found out that more than half of the organoids that were resistant to 5-FU monotherapy and sensitive to irinotecan maintained sensitive status to the treatment of 5-FU combined with irinotecan (combination-sensitive), while a quite proportion of them turn to become remarkably resistant to 5-FU plus irinotecan (combination-resistant). Some studies suggested the mechanism of 5-FU-based resistance was associated with the activation of some oncogenic pathways. However, the mechanism of the addition of 5-FU promoting the resistance to irinotecan is still unclear. Methods: To investigate the relationship between the combination-resistant organoids and corresponding patient responses, we collected patient clinical responses and survival outcomes. RNA sequencing and ATAC sequencing were performed to identify differential genes and their enrichment pathways. Moreover, small molecule inhibitors were used to try to reverse the resistance induced by 5-FU. Results: Twenty-one organoids showed sensitivity to irinotecan and resistance to 5-FU. Among them, 8 organoids (38.1%) were combination-resistant and 13 organoids (61.9%) showed sensitivity to combination treatment. Combination-resistant organoids corresponding patients had worse pathologic tumor regression grades (pTRGs) distribution (8/8, 100% had TRGs of 2-3 versus 100% had TRGs of 0-1 in combination-sensitive patients; p < 0.05) and disease-free survival (DFS) rates (HR:11.57, 95% IC:1.025-130.5; p = 0.0047) than patients whose organoids were combination-sensitive. Combination-resistant organoids had higher expression of Ki-67 and CD44 and reduced cleaved-caspase 3 in the combination treatment group and 5-FU group compared with the irinotecan group and control. Furthermore, we identified hedgehog pathway activation after adding 5-FU to irinotecan in combination-resistant organoids through RNA sequencing and ATAC sequencing. GANT-61, an inhibitor of the hedgehog pathway, increased sensitivity to the combination of 5-FU and irinotecan in combination-resistant organoids. Conclusions: We found that organoids which are sensitive to irinotecan turn to become resistant to 5-FU combined with irinotecan. Moreover, corresponding patients had worse responses and a lower DFS rate. The hedgehog pathway is activated in these organoids after the addition of 5-FU to irinotecan, and GANT-61 could reverse the resistance caused by 5-FU. However, more organoid-associated trials with large-scale patients are warranted to reverse the 5-FU-based resistance in the future.

High-dose radiation exposure induces gastrointestinal (GI) stem cell death, resulting in denudation of the intestinal mucosa and lethality from GI syndrome, for which there is currently no effective therapy. Studying an intestinal organoid-based functional model, we found that Sirtuin1(SIRT1) inhibition through genetic knockout or pharmacologic inhibition significantly improved mouse and human intestinal organoid survival after irradiation. Remarkably, mice administered with two doseages of SIRT1 inhibitors at 24 and 96 h after lethal irradiation promoted Lgr5+ intestinal stem cell and crypt recovery, with improved mouse survival (88.89% of mice in the treated group vs. 0% of mice in the control group). Moreover, our data revealed that SIRT1 inhibition increased p53 acetylation, resulting in the stabilization of p53 and likely contributing to the survival of intestinal epithelial cells post-radiation. These results demonstrate that SIRT1 inhibitors are effective clinical countermeasures to mitigate GI toxicity from potentially lethal radiation exposure.

Gallbladder carcinoma (GBC) is a lethal biliary tract malignant neoplasm. Patient-derived primary cancer cell lines (PDPCs) are appropriate models to explore biological characteristics and potential therapeutics; however, there is a lack of PDPCs in GBC. In this study, we aimed to establish and characterize the GBC PDPCs, and further investigated the intra-tumor heterogeneity (ITH). Multi-region sampling (3–9 regions) of the operable tumor tissue samples was used to establish PDPCs. Short tandem repeat genotyping for cell authentication and karyotyping was performed, followed by whole-exome sequencing and RNA sequencing to assess the ITH at the genetic and transcriptional levels, respectively. Thirty-eight PDPCs were successfully established from seven GBC patients and characterized. ITH was observed with a median of 38.3% mutations being heterogeneous (range, 26.6–59.4%) across all patients. Similar with other tumor types, TP53 mutations were always truncal. In addition, there were three genes, KMT2C, CDKN2A, and ARID1A, with truncal mutations in at least two patients. A median of 370 differentially expressed genes (DEGs) was identified per patient. Distinct expression patterns were observed between major histocompatibility complex (MHC) class I and II genes. We found the expression of MHC class II genes in the PDPC samples was closely regulated by CIITA, while that of MHC class I genes were not correlated with CIITA expression. The PDPCs established from GBC patients can serve as novel in vitro models to identify the ITH, which may pave a crucial molecular foundation for enhanced understanding of tumorigenesis and progression.

BACKGROUND Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths worldwide, but there is a shortage of effective biomarkers for its diagnosis. AIM To explore blood exosomal micro ribonucleic acids (miRNAs) as potential biomarkers for HCC diagnosis. METHODS T RESULTS The principal component analysis suggested that daily alcohol consumption could alter the blood exosomal miRNA profiles of hepatitis B virus positive non-HCC patients through miR-3168 and miR-223-3p. The miRNA profiles also revealed the tumor stages of HCC patients. High expression of miR-455-5p and miR-30c-5p, which significantly correlated with better overall survival in tumor tissues, could also be detected in blood exosomes. Two pairs of miRNAs (miR-584-5p/miR-106-3p and miR-628-3p/miR-941) showed a 94.1% sensitivity and 68.4% specificity to differentiate HCC patients from non-HCC patients. The specificity of the combination was substantially influenced by alcohol consumption habits. CONCLUSION This study suggested that blood exosomal miRNAs can be used as new non-invasive diagnostic tools for HCC. However, their accuracy could be affected by tumor stage and alcohol consumption habits.

BackgroundStatins are the most widely used drugs in elderly patients; the most common clinical application of statins is in aged hyperlipemia patients. There are few studies on the effects and mechanisms of statins on bone in elderly mice with hyperlipemia. The study is to examine the effects of atorvastatin on bone phenotypes and metabolism in aged apolipoprotein E-deficient (apoE–/–) mice, and the possible mechanisms involved in these changes.MethodsTwenty-four 60-week-old apoE–/–mice were randomly allocated to two groups. Twelve mice were orally gavaged with atorvastatin (10 mg/kg body weight/day) for 12 weeks; the others served as the control group. Bone mass and skeletal microarchitecture were determined using micro-CT. Bone metabolism was assessed by serum analyses, qRT-PCR, and Western blot. Bone marrow-derived mesenchymal stem cells (BMSCs) from apoE–/–mice were differentiated into osteoblasts and treated with atorvastatin and silent information regulator 1 (Sirt1) inhibitor EX-527.ResultsThe results showed that long-term administration of atorvastatin increases bone mass and improves bone microarchitecture in trabecular bone but not in cortical bone. Furthermore, the serum bone formation marker osteocalcin (OCN) was ameliorated by atorvastatin, whereas the bone resorption marker tartrate-resistant acid phosphatase 5b (Trap5b) did not appear obviously changes after the treatment of atorvastatin. The mRNA expression of Sirt1, runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), and OCN in bone tissue were increased after atorvastatin administration. Western blot showed same trend in Sirt1 and Runx2. The in vitro study showed that when BMSCs from apoE–/–mice were pretreated with EX527, the higher expression of Runx2, ALP, and OCN activated by atorvastatin decreased significantly or showed no difference compared with the control. The protein expression of Runx2 showed same trend.ConclusionsAccordingly, the current study validates the hypothesis that atorvastatin can increase bone mass and promote osteogenesis in aged apoE−/−mice by regulating the Sirt1–Runx2 axis.

Ambient fine particulate matter (PM

Hepatocellular carcinoma (HCC) is one of leading causes of cancer‐related deaths worldwide, but there is a shortage of effective biomarkers for its diagnosis. We aimed to explore exosomal miRNAs as potential biomarkers for HCC diagnosis. After the eliminations, this study included 89 patients from Xiangya Hospital of Central South University between August 2017 and May 2018. The principal component analysis (PCA) analysis suggested that daily alcohol consumption could alter the blood exosomal miRNA profiles of HBV positive non-HCC patients through miR-3168 and miR-223-3p. The miRNA profiles also revealed the tumor stages of HCC patients. High expression of miR-455-5p and miR-30c-5p, which significantly correlated with better overall survival in tumor tissues, could also be detected in blood exosomes. Two pairs of miRNAs (miR-584-5p/miR-106-3p and miR-628-3p/miR-941) showed 94.1% sensitivity and 68.4% specificity to differentiate HCC patients from non-HCC patients. The specificity of the combination was substantially influenced by alcohol consumption habits. This study suggested that blood exosomal miRNAs can be used as new non-invasive diagnostic tools for HCC. However, their accuracy could be affected by tumor stage and alcohol consumption habits.

The therapy of Huatanjiangqi capsule (HTJQ) has been used in the treatment of chronic obstructive pulmonary disease (COPD) clinically, with remarkable benefits. A lower functional activity of MRP1 is related to COPD development. However, the mechanism that contributes to MRP1 up-regulation by HTJQ was unclear. This study aimed to investigate the effects of HTJQ on the expression of Nrf2 and MRP1 in COPD rats. Rats were exposed to cigarette smoke plus lipopolysaccharide tracheal instillation to induce a COPD model. Then, Nrf2, HO-1, and MRP1 mRNA expression and their protein production in lung tissues were examined. The model group showed significant changes in the above assessments as compared to those of the control group (p < 0.05). The lung function of model group treated with low/high dose of HTJQ was significantly strengthened (both p < 0.05), and the inflammatory cells and goblet cells were decreased in lung tissues. Treatment with low/high dose of HTJQ resulted in obvious increase in Nrf2, HO-1, and MRP1 mRNA and protein expression levels (p < 0.05). HTJQ could up-regulate Nrf2 and MRP1 expression in lung tissues of COPD model rats. The mechanism by which HTJQ up-regulates MRP1 expression may be associated with Nrf2.

ObjectivesTo investigate whether the plasma level of trimethylamine N-oxide (TMAO), a proatherosclerotic intestinal microbiota metabolite, can be a predictor of ischemic brain injury secondary to carotid artery stenting (CAS).MethodsIn this multicenter, prospective cohort study, we enrolled patients with severe carotid artery stenosis (>70%) who were prepared for CAS. Plasma TMAO level was measured within 3 days before CAS, and MRI was performed 1 to 3 days after CAS.ResultsThe mean age of the 268 eligible patients was 64.4 years. New lesions on diffusion-weighted imaging (DWI) were detected in 117 patients (43.7%). TMAO level was higher in patients with new (DWI) lesions than in patients without new lesions (median 5.2 vs 3.2 µmol/L; p < 0.001). Increased plasma TMAO levels were associated with an increased risk of having new lesions on DWI after CAS (adjusted odds ratio for the highest vs lowest quartile, 3.85; 95% confidence interval, 1.37–7.56, p < 0.001; adjusted odds ratio for the third vs lowest quartile, 1.86; 95% confidence interval, 1.09–4.66, p = 0.02). The area under the receiver operating characteristic curve of TMAO was 0.706 for new lesions on DWI, and the optimal cutoff value was 4.29 µmol/L. The sensitivity, specificity, positive predictive value, and negative predictive value of TMAO levels ≥4.29 µmol/L for predicting new lesions on DWI were 61.5%, 74.8%, 65.5%, and 65.5%, respectively.ConclusionsIncreased TMAO levels are associated with an increased risk of new ischemic brain lesions on post-CAS MRI scans.

Hematite (α-Fe2O3) is a promising photoanode material for photoelectrochemical (PEC) water splitting due to its appropriate bandgap, good stability, and earth-abundance. However, the poor charge transfer property and sluggish kinetics of water oxidation limit the PEC performance of α-Fe2O3 photoanodes. Herein, a thin NiOx buffer layer was introduced between the Ti doped α-Fe2O3 (Fe2O3-Ti) layer and the fluorine-doped tin oxide (FTO) substrate without affecting the nanowire morphology and light absorption property of Fe2O3-Ti. This buffer layer can apparently suppress the charge recombination by mitigating the lattice mismatching between the Fe2O3-Ti film and the FTO substrate. In addition, the good conductivity of the NiOx film from the non-stoichiometric composition is also beneficial to the charge transfer. As a consequence, the photocurrent density was greatly improved by adding the NiOx layer in the Fe2O3-Ti photoanode, reaching 1.32 mA·cm−2 at 1.23 VRHE without any co-catalyst and sacrificial agent. This work gives a detailed analysis of the back contact in the hematite-based photoanode and provides an effective strategy for underlayer interface optimization.

In this paper, we investigate the effect of offering a flexible product in a newsvendor system with two products. We show that this flexible selling strategy can help firms effectively pool excess stocks to better match supply with demand and thus enhance profitability. However, offering a flexible product may also bring the potential risk of cannibalizing regular demand. We explore this trade-off by incorporating pricing decision for the flexible product when demand cannibalization exists. Our study shows that even when there is no demand induction effect, offering a flexible product still significantly improves profit. The value of offering a flexible product is highest when prices for specific products are the same, and it increases when the demands for specific products are more negatively correlated, more volatile, and more symmetric. Furthermore, the performance improvement is more salient when products have narrow profit margin or high overage risk.

The mantle transition zone (MTZ) bounded by physical discontinuities at 410 and 660 km depth is believed to play a key role in controlling mantle flow. Mineral physics experiments reveal that phase changes from olivine to β phase and γ -spinel to perovskite+magnesiowustite, at pressures equivalent to the existing at those mantle discontinuities, largely explain the variations in seismic velocity observed by the seismologists. The globally observed seismic velocity discontinuity at 660 km depth marks the bottom of the MTZ, which is the natural boundary between the upper and lower mantle. This discontinuity is considered an important mantle boundary, since it is especially associated with the mantle dynamics, the source of mantle plumes and the sinking of the subducted plates. On the other hand, temperature variations are the cause of thickening and thinning of the MTZ in correspondence with the positive and negative slopes of the Clapeyron curve at the 410 and 660 km mantle discontinuities, respectively. Receiver function analysis is a straightforward and commonly accepted method of studying the crust and upper-mantle structure through the use of teleseismic waveforms recorded at three-component seismic stations. This method requires the rotation of the ZNE displacement components to the radial and transversal components in the horizontal plane; and later the transformation of the radial and vertical components, which lie in the same vertical plane, into the L and Q components of the ground motion. By deconvolving the L component from the Q -component, both the source, far field path and instrument effects are removed. The resulting receiver function waveform is the ground’s impulse response. The analysis of each receiver function recorded at an array station provides, after moveout correction, a detailed estimation of the depth of the interface where the P-to-S seismic phase conversion is produced. However, the converted Ps phases at the discontinuities of 410 and 660 km depth are so weak that stacking of a large amount of waveforms is needed to enhance the signal-to-noise ratio in the receiver functions. In this study, we obtained 8600 teleseismic P-wave receiver functions recorded at 48 permanent broadband stations deployed in Yunnan and surroundings, which later were properly stacked in a single trace in 1°×1°-sized grid cells. Before stacking, the receiver functions were migrated from time domain to depth domain with the help of a reference earth model. Finally, we obtained 6 stacking images of receiver functions at latitude of 28°, 27°, 26°, 25°, 24° and 23°N, being the stacking depth in the 0–800 km range. The results indicate: (1) further north of 26°N, the average depths of the 410 and 660 km mantle discontinuities are in a range of 407–408 and 663–670 km, respectively, while the average thickness of the MTZ is within the range of 255–269 km, which are values very close to the global average thickness of 250 km. (2) South of 26°N, the average depths of the 410 and 660 km discontinuities are in a range of 412–426 and 675–703 km, respectively, and the average thickness of the MTZ is within the range of 262–279 km, which is obviously larger than the global average value of 250 km. The deepening of the 410 and 660 km discontinuities in the Yunnan region is obviously associated with the eastward subduction of the Indian plate below the Burma Arc; then, based on the greater thickness of the MTZ beneath the Yunnan region, we suggest that the eastward subduction of the Indian plate occurs mainly south of 26°N in the Yunnan region.

More frequent extreme heat conditions, forecasted in the context of climate change, are likely to significantly affect microbial communities that are important for nitrogen (N) cycling and N 2 O emissions. Here we report an incubation study on the effect of a range of temperatures (20, 30 and 40 °C) on the growth of bacteria (ammonia oxidizing bacteria, AOB), archaea (ammonia oxidizing archaea, AOA), fungi (18SrRNA) and key denitrifying communities ( nirK- , nirS- and nosZ -denitrifiers), and effects on N 2 O emissions, following the application of urea and an organic manure in an acidic soil. Results showed that the growth of ammonia oxidizers and denitrifiers were strongly inhibited by the higher temperatures, particularly at 40 °C. AOB grew well with the application of urea and manure but AOA grew in the Control at 20 °C. Bacterial denitrifiers only grew mostly at 20 °C and did not grow at the higher temperatures. In contrast, fungi communities, including fungal nirK -type communities, grew under all three temperatures in the manure treatment, but not in the urea treatment, showing fungal tolerance to high temperatures and an interaction between nitrogen (N) source and fungal growth. N 2 O emissions increased in the urea treatments and decreased in the manure treatments with increasing temperature, possibly pointing to different mechanisms of N 2 O production in the urea and manure treated soils. These results suggest that different microbial communities will respond differently to extreme heat conditions and the type of N source applied will also have an interactive effect with temperature on the microbial growth and N 2 O emissions.

The long-term contamination of soil by microplastics may pose risks that are often still not well understood, and the ecological effects of microplastics are mainly dependent on their environmental behavior in environments. This study used saturated quartz sand as a solid porous medium to study the migration and influencing factors of 40–48 μm polyethylene (PE) particles in saturated porous media. The breakthrough curves at different injection concentrations (0.3, 0.4, 0.5 mg/L), flow rates (1.0, 1.5, 2.0, 2.5 ml/L), porous medium particle sizes (1–2, 2–4 mm), ionic strengths (0, 0.01, 0.05 mol/L) and concentrations of fulvic acid (FA) (0, 5, 10 mg/L) were compared and analyzed. The Derjaguin-Landau-Verwey-Overbeek (DLVO) theory was used to more accurately explain relevant transport behaviors. The results showed that the input concentration, flow rate, and particle size can affect the migration of PE particles individually or in combination. As ionic strength increased, the repulsion between microplastics and quartz sand gradually disappeared according to DLVO theory, and their attraction gradually strengthened. As a result, fewer microplastics could penetrate the sand column and reach the water body. With the continuous addition of FA, the repulsive energy between microplastics and quartz sand rose from DLVO theory, and the migration ability of microplastics initially increased before becoming stable because of the effect of straining. In all cases, the migration ability of PE was low (C/C0

Multidrug resistance-related protein 1 (MRP1) is involved in the biological transport of several molecules with diverse structural characteristics outside of the cell. In addition to its transport activity, MRP1 exhibits multiple defense mechanisms in vivo. MRP1 is highly expressed in normal lung tissues and plays a protective role in the process of chronic obstructive pulmonary disease. In the present study, human bronchial epithelial cells (16HBE14o-cells) were stimulated by cigarette smoke extract (CSE) in vitro to simulate a smoking environment. On this basis, the mechanism of Allyl isothiocyanate (AITC) administration on the expression of MRP1 in CSE-stimulated 16HBE14o-cells was investigated. The effects of CSE on the viability of 16 HBE14o-cells were investigated by an MTT assay. The changes in the mRNA expression levels of nuclear erythroid factor 2 (Nrf2) and MRP1 were investigated in CSE-stimulated 16HBE14o-cells using western blotting and reverse transcription quantitative PCR (RT-qPCR). Immunofluorescence analysis was used to detect Nrf2 nuclear translocation. Incubation of the cells with 5% CSE for 24 h had minor effects on cell viability and resulted in the activation of the JNK and p38MAPK signaling pathways. AITC activated the JNK pathway, inhibited the activation of the p38MAPK pathway in 16HBE14o-cells stimulated by 5% CSE and upregulated the expression levels of Nrf2 and MRP1 in a time-dependent manner. The upregulation of Nrf2, MRP1 and of Nrf2, and MRP1 mRNA expression levels in CSE-stimulated cells was inhibited by pretreatment with SP600125 (a JNK pathway inhibitor). Furthermore, the fluorescence intensity in the nucleus was significantly enhanced following AITC pretreatment and the analysis indicated nuclear translocation of Nrf2 in the cells. These results indicated that Nrf2 and MRP1 expression levels in CSE-stimulated cells were altered following AITC pretreatment. Thus demonstrating that the primary mechanism may be associated with activation of the JNK pathway, while the p38MAPK pathway may not be involved.

Taste loss is one of the debilitating complications in radiation-induced oral mucositis (RIOM), as occurs in head and neck cancer patients undergoing radiotherapy. We report here a radio-mitigation effect of Sirtuin 1 (SIRT1) inhibitors in taste bud organoids and a mouse model of radiation-induced taste bud injury. The organoids, developed from circumvallate (CV) papilla, were irradiated with single dose of X-rays and inhibitors of SIRT1 or SIRT2 were added 24 h later. The survival was evaluated by measuring the number and size of regenerated organoids after irradiation (IR). Oral mucositis (OM) was induced by IR of the oral region of Lgr5-lacZ transgenic mice. The surviving Lgr5+ taste bud stem cells were identified after lacZ-staining and the mucosal ulceration on tongue dorsal surface was determined by histological methods. Results showed that SIRT1 inhibitors (nicotinamide, EX527, salermide and sirtinol), but not SIRT2 inhibitors, significantly improve taste bud organoid survival after IR. Remarkably, administration of nicotinamide (NAM), a recognized inhibitor of SIRT1 to mice 24 h after IR promotes the survival of Lgr5+ taste bud stem cells, resulting in alleviated tongue mucositis. In conclusion, SIRT1 inhibitors promote Lgr5+ taste bud stem cell survival and mitigate RIOM in mice. These observations have important implications for efforts to develop therapeutic strategies against taste dysfunction and mucosal ulceration in RIOM.

Tissue sampling of biliary tract carcinomas (BTCs) for molecular characterization is challenging. The aim of this study was to investigate the possibility of identifying individual actionable mutations derived from bile cell-free DNA (cfDNA) using targeted deep sequencing. Ten BTC patients, four with gallbladder carcinomas and six with cholangiocarcinomas, were enrolled in the present study. Using targeted deep sequencing with a panel of 150 tumor-related genes, paired bile cfDNA and tumor DNA were analyzed for mutational variants individually and then compared. The present study, to the best of our knowledge, is the first to reveal that bile cfDNA is predominantly comprised of long DNA fragments, which is not the case for plasma cfDNA. Herein, paired bile cfDNA and tumors from ten BTC patients were examined using targeted deep sequencing. When comparing bile cfDNA and tumor DNA for single nucleotide variation (SNV)/insertion and deletion (Indel), the results using targeted deep sequencing revealed high sensitivity (94.7%) and specificity (99.9%). Additionally, the sensitivity of detecting a copy number variation (CNV) was 75.0%, with a specificity of 98.9%. When comparing two bile extraction methods, including percutaneous transhepatic cholangial drainage and operation, no significant difference in SNV/Indel or CNV detection sensitivity was noted. Moreover, when examining the tumor stage and incidence site, AJCC stage II and the distal bile duct both had significantly decreased CNV detection sensitivities. The present study revealed that targeted deep sequencing can reliably detect mutational variants within bile cfDNA obtained from BTC patients. These preliminary results may shed light on bile cfDNA as a promising liquid biopsy for BTC patients.

Background: Colon signet-ring cell carcinoma (SRCC) is a rare type of malignant dedifferentiated adenocarcinomas, and associated with poor survival. However, an in-depth study of the biological features of colon SRCC is hampered by lack of in vitro model. Our study was designed to establish and characterize cell cultures from SRCC for further translational and biological research. Methods: By harnessing the power of organoid culture system, we established a human colon SRCC organoid line from surgical sample of one patient with colon SRCC. The organoid line was characterized at the morphology, histology, ultrastructure, chromosome stability level and its tumorigenic ability was verified by xenografts. Capture-based targeted DNA sequencing combining with drug screen based on a bespoke 88 compound library identified the possible target and the treatment response was confirmed by in vivo organoid system and in vitro xenografts. Findings: Our data showed that the colon SRCC organoid line, YQ-173, resemble histological and growth characteristics of the original tumor cells and have high tumor formation rate. DNA sequencing indicated JAK2 might be the treatment target. In vitro drug screen found that AT9283 and Pacritinib could be the effective JAK2 inhibitors, which was consistent with in vivo xenografts' response. Interpretation: We report, for the first time, the establishment of SRCC organoid line allowing in-depth study of SRCC biology, as well as a strategy to assess in vitro drug testing in a personalized fashion. Funding: This work was supported by National Natural Science Foundation of China (31470826, 31670858, 81672374), Science and Technology Commission of Shanghai Municipality (16411966300), and Shanghai Sailing Program (19YF1409500). Declaration of Interest: The authors declare no potential conflicts of interest. Ethical Approval: The tissue was obtained with informed consent for experimentation with the patient and this study was approved by the ethical committee of Fudan University Shanghai Cancer Center. All animal experiments were performed according to the guidelines for the care and use of laboratory animals and were approved by Institutional Animal Care and Use Committee of Fudan University.

The increasing vehicle numbers in China have raised issues on effective mitigation of the vehicular NOx emissions recently. Notably, temporally growing and spatial agglomeration of high vehicular NOx emissions make an essential challenge to the mitigation strategy-makers. However, so far, there have been few studies to give insight into the socioeconomic drivers like the spatial imbalance of socioeconomic development, vehicle structure and road infrastructure to help governors. To fill the above gap, this study explores drivers of temporal change and spatial differences by building a temporal-spatial decomposition model and accounting for national and regional NOx emissions from vehicles in China from 2005 to 2015. Results show that, of all the driving forces in this study, only road vehicle carrying capacity (ΔNVI) acts as a primary driving force for both temporal growing and spatial agglomeration of vehicular NOx emissions in China. Regional vehicle emission intensity (ΔNNI) and road economic growth (ΔNEI) only mainly contributed to spatial agglomeration. While economic development (ΔNG) played a crucial role in the temporal growing of vehicular NOx emissions in China. These findings indicate that the future mitigation policy should fully cover the comprehensive socioeconomic factors, which would be useful for China and other developing countries when aiming to improve the performance of their current vehicle emissions policy system.

Microbes play a key role in soil nutrient cycling and supply in the extensive semiarid grassland ecosystem, where no fertilizers are applied. However, the role of fungi vs bacteria in nitrogen (N) mineralization in such ecosystem is poorly understood. The objective of this study was to determine the impacts of different mowing practices on fungal and bacterial communities and the relationships between the two microbial communities and net N mineralization rate (R m). This study was based on a 13-year mowing experiment in Inner Mongolia. The treatments included mowing once every second year (M1/2), mowing twice every 3 years (M2/3), mowing once a year (M1), mowing twice a year (M2), and the unmown (control, CK). Soil basic chemical properties, R m, microbial biomass, bacterial and fungal community abundance, and diversity were determined, and fungal phylogeny and the relationship between microbial community and R m were analyzed. Moderate mowing (M1/2, M2/3, and M1) enhanced soil carbon and nitrogen stocks, R m, fungal community abundance and diversity which might mainly because of the increased decomposer fungi species, but the higher frequency mowing (M2) significantly decreased the above. There was a significant correlation between fungal community abundance and R m (r = 0.688, P = 0.005) in this study. However, different mowing practices had little effect on the bacterial community, which might due to human disturbance (mowing practices) and poor environmental conditions (drought, limited available nitrogen and phosphorus). Overall, fungi may play a more important role than bacteria in N mineralization under mowing management in such a grassland ecosystem. Moderate-frequency mowing, e.g., M1, is more appropriate for maintaining soil microbial communities and soil fertility, whereas the high-frequency mowing, e.g., M2, is not sustainable for maintaining soil nutrients and microbial community in such a semiarid grassland ecosystem in a long term.

Methane (CH4) is a potent greenhouse gas, and soil can both be a source and sink for atmospheric CH4. It is not clear how future climate change may affect soil CH4 emissions and related microbial communities. The aim of this study was to determine the interactive effects of a simulated warmer and drier climate scenarios and the application of different nitrogen (N) sources (urea and manure) on CH4 emissions and related microbial community abundance in a vegetable soil. Greenhouses were used to control simulated climate conditions which gave 2.99 °C warmer and 6.2% lower water content conditions. The field experiment was divided into two phases. At the beginning of phase II, half of the greenhouses were removed to study possible legacy effects of the simulated warmer and drier conditions. The responses in methanogen and methanotroph abundance to a simulated climate change scenario were determined using real-time PCR. The results showed that the simulated warmer and drier conditions in the greenhouses significantly decreased CH4 emissions largely due to the lower soil moisture content. For the same reason, CH4 emissions of treatments in phase I were much lower than the same treatments in phase II. The abundance of methanotrophs showed a more significant response than methanogens to the simulated climate change scenario, increasing under simulated drier conditions. Methanogenic community abundance remained low, except where manure was applied which provided a source of organic C that stimulated methanogen growth. Soil moisture content was a major driver for methanotroph abundance and strongly affected CH4 emissions. The application of N source decreased CH4 emissions probably because of increased methanotrophic activity. CH4 emissions were positively correlated to methanogenic abundance and negatively correlated to methanotrophic abundance. These results demonstrate that projected future climate change conditions can have a feedback impact on CH4 emissions from the soil by altering soil conditions (particularly soil moisture) and related microbial communities.

Nitrous oxide (N2O) is a potent greenhouse gas and is mainly produced from agricultural soils especially vegetable soils with large N fertilizer input. How future projected climate change may impact on the N2O emissions and the related key (de)nitrifier communities in such ecosystem is poorly understood. The aim of this field study was to determine the interactive effects of a simulated warmer and drier climate on (de)nitrifier communities and N2O emissions in a vegetable soil. A warmer (+3.3 °C) and drier climate (−14.4% soil moisture content) was created with greenhouses with or without urea N fertilizer application. The variation of microbial population abundance and community structure of Ammonia-oxidizing archaea (AOA), bacteria (AOB) and denitrifiers (nirK/S, nosZ) were determined using Real time-PCR and sequencing. The results showed a strong interactive effect of simulated climate change with N fertilizer applications, whereby the impacts of warmer and drier conditions on the microbial communities and N2O emissions were more evident when N fertilizer was applied. The simulated warmer and drier conditions in the greenhouses significantly decreased N2O emissions largely due to the drier soil conditions. The abundance and community structure of AOB showed more rapid responses than AOA under the simulated climate conditions when N fertilizer was applied. Changes of AOB community structure were significantly correlated with soil moisture content and NH4+-N concentration. The simulated climate change did not affect the nirS gene abundance, but significantly increased nirK gene abundance, and significantly decreased nosZ gene abundance with urea application. N2O emissions were positively correlated with the bacterial amoA abundance and with the ratio of nirK/nosZ gene abundance. Therefore, bacterial amoA, nirK- and nosZ-type denitrifiers are the dominant microbial communities which were affected by the simulated climate conditions and are thus critically important for N cycling in vegetable soils under a changing climate.

We study the pricing problem for a firm with two servers where heterogeneous customers can choose between deterministic service and probabilistic service. We find that different queueing priority policies do not affect the firms revenue but affect the firms optimal pricing strategies. Specifically, when the flexible customers (who choose probabilistic service) have a high priority, the optimal price of the deterministic service could be lower than the one of the probabilistic service in a small or moderate market.

The feedback of greenhouse gas emissions to climate change is vital for understanding and predicting the impact of global warming on ecosystem functions. However, the legacy effect of simulated climate change on nitrous oxide (N2O) emissions and the associated microbial guilds remain largely unknown. Using a climate change field-based mesocosm facility, we studied the impact of a warmer and drier environment and its legacy effect on in situ N2O emissions with different fertilizer types (manure and urea). The related functional guilds including N2O-producer [ammonia-oxidizing archaea and bacteria (AOA and AOB), nirS/K-type denitrifying bacteria, and nirK-type denitrifying fungi] and N2O-reducer (nosZI and nosZII) were analyzed by using high throughput and cloning sequencing. The simulated climate change significantly decreased in situ N2O emissions in the fertilized soil (urea- or manure-treated) while increasing the emissions in a non-fertilized soil. The AOA and AOB were well adapted to the simulated climate change condition in the manure- and urea-treated soil, respectively. In contrast, the fungal nirK-type N2O-producers were well adapted in non-fertilized soil. The abundance of nosZII was significantly stimulated by simulated climate change in both fertilized and non-fertilized soils. Moreover, different fertilizer types modulated the resilience of the microbial guilds. The AOA and the nirS-type denitrifying bacteria showed strong resilience, leading to a significant increase of N2O emissions in the manure-treated soil. The strong resilience was also observed in nosZII clade N2O-reducers, and the abundance of the species related to Candidatus Promineofilum breve and Gemmatirosa kalamazoonesis was stimulated by the legacy effect of simulated climate change in the urea-treated soil. The in situ N2O emissions were negatively correlated to nosZII rather than nosZI. These results highlight a significant potential of nosZII in mitigating N2O emissions under a projected climate change, especially in agroecosystems, where a large amount of fertilizers is commonly used.

Ambient fine particulate matter (PM2.5) can cause adverse health effects through the generation of reactive oxygen species (ROS) after inhalation. Humic-like substances (HULIS) are major constituents contributing to the ROS-generation potential in organic aerosols. In this study, PM2.5 samples in urban Shanghai during autumn and winter (2018-2019) were collected. Mass-normalized ·OH generation rate in surrogate lung fluid (SLF) was used to denote the intrinsic ROS-generation potential of PM2.5 or of the HULIS isolated from PM2.5. In this study, ROS-generation potential of PM2.5 decreased with increasing ambient PM2.5 concentration due to higher percentage of inorganic components in high PM2.5 event. Same trend was observed for the ROS-generation potential of unit mass of HULIS, which was higher when HULIS and PM2.5 concentrations were both relatively lower. The HULIS with high ROS-generation potential but low concentration (High-ROS/Low-Conc HULIS) were likely produced by the atmospheric aqueous-phase reactions during nighttime or under high relative humidity conditions, not from biomass burning emissions or the photochemical pollution products. The association between ROS-generation potential and light absorption properties of HULIS was studied as well. The High-ROS/Low-Conc HULIS also showed stronger light absorbance than the other HULIS. Our results implied the potentially important roles that HULIS species might play in atmospheric environment and human health even when the PM2.5 pollution is low.

This study aims to understand the characteristics of surface ozone (O3), search for factors affecting the variations in its concentration, and estimate its impacts on the secondary organic carbon (SOC) levels and atmospheric oxidation capacities in the Yangtze River Delta (YRD). Four years of continuous observations (2014–2017) of the surface O3, organic carbon, elemental carbon, nitrogen oxides, PM2.5 and meteorological factors along with three years of measurements (2015–2017) of the concentrations of 56 volatile organic compounds were conducted at a rural site. Our measurements showed that the total number of O3 pollution days more than doubled over the four-year period, from 28 days in 2014 to 76 days in 2017. The annual mean of the maximum daily 8-h average O3 concentration during the months with the strongest solar radiation (July–September) showed a 6.8% growth rate, from 124.5 (2014) to 149.8 μg m−3 (2017). Regional transport was shown to be the dominant contributor to the high level of O3 based on a process analysis of the O3 variation using the Weather Research and Forecasting-Community Multiscale Air Quality model for this site. The simulation results indicated that the city junction site served well as an epitome of the regional background of the YRD. We also found that the level of SOC, which is a major component of PM2.5 that results from atmospheric oxidizing processes, gradually increased with the increase in the surface O3 level, even though the overall PM2.5 concentration significantly decreased each year. There was an increasingly strong correlation between SOC and Ox (O3 + nitrogen dioxide) during both the daytime and night-time from 2014 to 2017 when the highest annual O3 concentration was observed. These findings imply that the atmospheric oxidation capacity increased and likely contributed to the SOC production in the YRD during 2014–2017.

Purpose: Biliary tract cancer (BTC) is a kind of malignant digestive system tumors without a targeted drug approved in the clinic. A high proportion of gene mutations and abnormal expressions of ERBB pathway have been reported in BTC. However, there are few studies on targeting ERBB in BTC. In this study, we aim to explore the role of targeting ERBB and its potential resistance mechanisms in BTC. Materials and Methods: We used the tumor tissue samples of BTC patients to establish patient-derived cell lines (PDCs), and perform the biological characteristics of PDCs. At the first, Western Blot was used to select the PDCs with different ERBB2 expressions as in vitro models. Secondly, colony formation assay was to verify the sensitivity of inhibitors and Western blot was to confirm the change of signaling pathways in PDCs. Then, whole exome sequencing (WES) and transcriptome sequencing (RNA-seq) were integrated to explore the potential resistance mechanisms of ERBB inhibitors. Moreover, stable PDCs with overexpression and knockdown established by lentiviral vectors were used. Finally, PDC xenografts were used to further confirm the hypothesis from the in vitro study. Results: Twenty-two BTC PDCs were successfully established in this study. According to ERBB2 expression, nine PDCs were selected for the sensitivity evaluation of ERBB inhibitors. Four PDCs of them are sensitive to three ERBB inhibitors, while the other five PDCs are not. Furthermore, colony formation assay has demonstrated the difference of colonies between sensitive cell lines and resistant cell lines for all three ERBB inhibitors. Meanwhile, Western Blot showed that inhibition activity is negatively correlated with phosphorylation of ERBB2 in PDCs. These results demonstrated that decreasing ERBB activity could inhibit the proliferation of BTC PDCs. Further analysis showed that all four sensitive PDCs highly expressed ERBB2, while four resistant PDCs carried with low ERBB2 expression levels. HCC783 was the only one PDC with high ERBB2 expression as well as resistance to ERBB inhibitors and was considered as a primary resistant PDC for follow-up study. RNA-seq analysis results showed that SMARCA1 was significantly lower (p < 0.01) and the gene transcriptional spectrum were greatly changed in resistant PDCs compared with sensitive PDCs, suggesting that SMARCA1 may have been involved in resistance mechanisms of ERBB inhibitors in BTC. Importantly, stable PDCs were established with overexpression and knockdown of SMARCA1 and the colony formation assay with them confirmed this hypothesis in vitro. The results demonstrated that a resistant cell line by overexpression of SMARCA1 could make sensitive to ERBB inhibitors, while a sensitive cell line by knockdown of SMARCA1 could become relatively resistant to ERBB inhibitors. The exploration of resistance mechanisms triggered by SMARCA1 and the in vivo validation with PDC xenografts are undergoing. Conclusions: In this study, the PDC model, drug screening and colony formation assay were integrated to verify that ERBB inhibitors could prevent the proliferation of BTC cells for the first time. Furthermore, WES and RNA-seq combined with stable PDC models were used to discover and validate that SMARCA1 is one of the potential resistance mechanisms of ERBB inhibitors in BTC. Citation Format: Feiling Feng, Qingbao Cheng, Dadong Zhang, Chen Liu, Xiaoya Xu, Bin Li, Huizhen Wang, Yong Yu, Bin Li, Xiaobing Wu, Jun Zhou, Kaijian Chu, Zhenghua Xie, Qingxiang Gao, Lei Xiong, Fugen Li, Bin Yi, Xiaoqing Jiang. Study on the role of targeting ERBB and its potential resistance mechanism in biliary tract cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1912.

Purpose: Lung cancer is the most common malignancy with the highest morbidity and mortality worldwide. Low-dose computed tomography (LDCT) is one of the main tools for lung cancer screening and diagnosis. However, high false positive, high cost, over-diagnosis and radiation exposure are major drawbacks to screen pulmonary nodules by LDCT. Thus, a noninvasive diagnostic approach with high specificity and accuracy is badly needed to enhance LDCT. Liquid biopsy represent a valuable non-invasive approach when biopsy or resection is not the first choice. Till now, it is rare to see the studies on exosome-derived miRNAs as early diagnosis biomarkers to distinguish benign and malignant pulmonary nodules using small RNA sequencing. Here, we aimed to explore the diagnostic value of a panel of significantly differential expressed plasma exosomal miRNAs between benign and malignant pulmonary nodule samples in Chinese cohorts. Materials and Methods: This study was registered at Chinese Clinical Trial Registry (www.chictr.org.cn) with registration number ChiCTR1800019877. Forty-five patients including twenty-six lung adenocarcinoma and nineteen benign nodules with various pathological characteristics were enrolled as a training cohort. A test cohort consisted of sixty-two patients with twenty-four benign nodules patients similar to training cohort and thirty-eight lung adenocarcinoma. Exosomes were precipitated from the plasma, and small RNA sequencing was performed to identify the differential expressed miRNAs. A statistical model consisting of a panel of exosomal miRNAs was trained to discriminate benign nodules from cancerous ones. The model was validated in the independent test cohort, and re-confirmed in an external dataset from another Chinese cohort. Enriched Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were also performed. Results: Characteristic proteins and morphology of exosomes were characterised by western blotting, nanoparticle tracking analysis and scanning electron microscopy. Five differential expressed miRNAs (let-7b-3p, miR-101-3p, miR-125b-5p, miR-150-5p, and miR-3168) with median expression > 50 were selected by LASSO-penalized regression as a linear model to classify samples into benign or maligant groups with 10-fold cross-validation to determine the model parameters. When the specificity set 94.7% and 91.7% for the training and test cohorts, respectively, the model had 57.7% and 57.9% sensitivity in both cohorts. The model was also confirmed in an external dataset with 87.5% specificity and 53.1% sensitivity. The expression of each biomarker in benign, adenocarcinoma in situ/microinvasive adenocarcinoma and invasive adenocarcinoma nodules were gradually altered. Four of the five biomarkers were gradually increased, whereas one miRNA was gradually decreased. GO and KEGG analysis demonstrated that biological process and pathways of the genes targeted by five biomarkers were associated with tumor development. Conclusions: This study using small RNA sequencing identified five plasma exosome-derived differentially expressed miRNAs as a diagnosis model to distinguish benign and malignant pulmonary nodules, which provides insights into the feasibility of exosomal miRNAs as a novel early diagnosis approach for lung adenocarcinoma. Citation Format: Di Zheng, Yang Yang, Chunyan Wu, Huizhen Wang, Jiyang Zhang, Shiyi Liu, Xiaoya Xu, Hao Chen, Dadong Zhang, Fugen Li, Jian Ni, Gening Jiang, Jianfang Xu. A panel of plasma exosomal miRNAs as diagnosis biomarker to distinguish benign and malignant nodules in non-small cell lung cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 756.

Background Dachshund homologue 1 (DACH1) is highly expressed in LGR5+ intestinal stem cells and colorectal tumours. However, the roles of DACH1 in intestinal cell stemness and colorectal tumorigenesis remain largely undefined. Methods We used immunohistochemistry, western blotting and quantitative real-time PCR to analyse DACH1 expression in colorectal cancer (CRC) samples. CRISPR/Cas9 gene editing and lentiviral vector-mediated overexpression and shRNA-mediated knockdown of DACH1 were utilized to modulate DACH1 expression in cell lines and organoids. An intestinal organoid-based functional model was analysed, and cancer cell colony formation, sphere formation assays and murine xenotransplants were performed to reveal the role of DACH1 in CRC cell proliferation, stemness and tumorigenesis. Immunofluorescence, co-immunoprecipitation, RNA interference and microarray data analyses were conducted to demonstrate the association between DACH1 and the bone morphogenetic protein (BMP) signalling pathway. Findings DACH1 is specifically expressed in discrete crypt base cells, and increased DACH1 expression was found in all stages of CRC. Moreover, the high expression of DACH1 independently predicted poor prognosis. In colon cancer cells, shRNA-mediated suppression of DACH1 inhibited cell growth in vitro and in vivo. By studying the intestinal organoid-based functional model, we found that depletion of DACH1 reduced the organoid formation efficiency and tumour organoid size. DACH1 overexpression stimulated both colonsphere formation and tumour organoid formation in the context of dysregulated BMP signalling. Mechanistic characterizations indicated that overexpression of DACH1 affects a subset of stem cell signature genes implicated in stem cell proliferation and maintenance through the suppression of BMP signalling via SMAD4. Interpretation Together, our study highlights DACH1 as an integral regulator of BMP signalling during intestinal tumorigenesis, and DACH1 could be a potential prognostic marker and therapeutic target for colorectal cancer patients.

Aims Chronic obstructive pulmonary disease (COPD) is a disease with high morbidity and mortality worldwide, which can cause serious social and economic burdens. Allyl isothiocyanate (AITC) is one of the most common natural isothiocyanates and has been shown to have anti-inflammatory and antioxidant biological activities. The purpose of this study was to investigate whether AITC regulated Multidrug resistance-associated protein 1 (MRP1), reactive oxide species (ROS) and reduced glutathione (GSH) levels via Nrf2 and Notch1 signaling pathways to treat COPD and whether there was an interaction between these two pathways. Main methods Lung function indexes and histopathological changes in mice were determined by lung function instrument and HE staining, respectively. The protein expression was analyzed using immunohistochemistry and Western blotting. The mRNA expression was measured by RT-PCR in human bronchial epithelial cell line 16HBE. The contents of ROS, GSH and GSSG were detected by kits in 16HBE cells. Key findings The protein expression of Notch1, Hes1, MRP1, Nrf2, and HO-1 in lung tissues of WT mice and untransfected cells were significantly down-regulated in COPD, then significantly ameliorated in treatment groups. The protein expression of MRP1, Notch1 and Hes1 in lung tissues of Nrf2−/− mice were markedly reduced. There was a significant reduction in expression of Nrf2, HO-1 and MRP1 in si-Notch1 transfected cells. Pretreatment with AITC markedly improved oxidative stress and GSH-redox disorder in COPD. Significance Our study demonstrates that there is a potential interaction between Nrf2 and Notch1 signaling pathways during treatment of COPD.

This paper analyses the function expression of the optimal minimum competition window which integrates the network node number, average collision data frame length and sending rate of data frame. At the same time, the proposed optimised method of MAC protocol combines with the SVN neural network which can memory communication environment which include node density and mobile velocity. Each terminal node in the network runs proposed MAC protocol optimisation algorithm based on this function expression to adaptive adjust their minimum competition window and back off the optimal value to improve the network performance. The simulation results show that the effort of optimised algorithm in the Ad Hoc system is limited for unsaturated business VANET; but high accuracy and effect of the optimised algorithm in aspects of throughput and transmission delay has improved significantly for saturated business of VANET.

Accumulating evidence indicates that patient-derived organoids (PDOs) can predict drug responses in the clinic, but the ability of PDOs to predict responses to chemoradiation in cancer patients remains an open question. Here we generate a living organoid biobank from patients with locally advanced rectal cancer (LARC) treated with neoadjuvant chemoradiation (NACR) enrolled in a phase III clinical trial. Our co-clinical trial data confirm that rectal cancer organoids (RCOs) closely recapitulate the pathophysiology and genetic changes of corresponding tumors. Chemoradiation responses in patients are highly matched to RCO responses, with 84.43% accuracy, 78.01% sensitivity, and 91.97% specificity. These data imply that PDOs predict LARC patient responses in the clinic and may represent a companion diagnostic tool in rectal cancer treatment.