Nitrogen removal performance of improved subsurface wastewater infiltration system under various influent carbon-nitrogen ratios.

Subsurface wastewater infiltration system (SWIS) has been recognized as a simple operation and environmentally friendly technology for wastewater purification. However, effectively removing nitrogen (N) remains a challenge, hindering the widespread application of SWIS. In this study, zero-valent iron (ZVI) and porous mineral material (PMM) were applied in SWIS to improve the soil matrix. Our results suggested that the addition of ZVI and PMM could simultaneously enhance N removal efficiency and reduce nitrous oxide emissions. This could be attributed to the abundant electrons generated by ZVI alleviating the electronic limitation of denitrification and the porous structure of PMM providing solid phase support for microbial growth. In addition, the abundance of microbial functional genes increased in modified SWIS, which could further explain the higher pollutant removal efficiency. Overall, this study provides new insights into the mitigation of wastewater pollution and greenhouse gas emissions in SWIS. PRACTITIONER POINTS: ZVI and PMM can adapt to different C loads and enhance pollutant removal efficiency in SWIS. Increasing C-N ratios positively affected the nitrate removal performance and negatively affected ammonium removal performance in SWIS. The amending soil matrix promoted the reduction of the N2 O to N2 and greenhouse gas emissions were well controlled. The abundance of microbial functional genes increased with the improvement of the soil matrix.

Large Seasonal Variation in Nitrogen Isotopic Abundances of Ammonia Volatilized from a Cropland Ecosystem and Implications for Regional NH3 Source Partitioning.

Ammonia (NH3) volatilization from agricultural lands is a main source of atmospheric reduced nitrogen species (NHx). Accurately quantifying its contribution to regional atmospheric NHx deposition is critical for controlling regional air nitrogen pollution. The stable nitrogen isotope composition (expressed by δ15N) is a promising indicator to trace atmospheric NHx sources, presupposing a reliable nitrogen isotopic signature of NH3 emission sources. To obtain more specific seasonal δ15N values of soil NH3 volatilization for reliable regional seasonal NH3 source partitioning, we utilized an active dynamic sampling technique to measure the δ15N-NH3 values volatilized from maize cropping land in northeast China. These values varied from -38.0 to -0.2‰, with a significantly lower rate-weighted value observed in the early period (May-June, -30.5 ± 6.7‰) as compared with the late period (July-October, -8.5 ± 4.3‰). Seasonal δ15N-NH3 variations were related to the main NH3 production pathway, degree of soil ammonium consumption, and soil environment. Bayesian isotope mixing model analysis revealed that without considering the seasonal δ15N variation in soil-volatilized NH3 could result in an overestimate by up to absolute 38% for agricultural volatile NH3 to regional atmospheric bulk ammonium deposition during July-October, further demonstrating that it is essential to distinguish seasonal δ15N profile of agricultural volatile NH3 in regional source apportionment.

Ticks (Acari: Ixodidae) infesting cattle in coastal Kenya harbor a diverse array of tick-borne pathogens.

Ticks and the microbes they transmit have emerged in sub-Saharan Africa as a major threat to veterinary and public health. Although progress has been made in detecting and identifying tick-borne pathogens (TBPs) across vast agroecologies of Kenya, comprehensive information on tick species infesting cattle and their associated pathogens in coastal Kenya needs to be updated and expanded. Ticks infesting extensively grazed zebu cattle in 14 villages were sampled and identified based on morphology and molecular methods and tested for the presence of bacterial and protozoan TBPs using PCR with high-resolution melting analysis and gene sequencing. In total, 3,213 adult ticks were collected and identified as Rhipicephalus appendiculatus (15.8%), R. evertsi (12.8%), R. microplus (11.3%), R. pulchellus (0.1%), Amblyomma gemma (24.1%), A. variegatum (35.1%), Hyalomma rufipes (0.6%), and H. albiparmatum (0.2%). Ticks were infected with Rickettsia africae, Ehrlichia ruminantium, E. minasensis, Theileria velifera and T. parva. Coxiella sp. endosymbionts were detected in the Rhipicephalus and Amblyomma ticks. Co-infections with two and three different pathogens were identified in 6.9% (n = 95/1382) and 0.1% (n = 2/1382) of single tick samples, respectively, with the most common co-infection being R. africae and E. ruminantium (7.2%, CI: 4.6 - 10.6). All samples were negative for Coxiella burnetii, Anaplasma spp. and Babesia spp. Our study provides an overview of tick and tick-borne microbial diversities in coastal Kenya.

Transformation mechanisms of ammonium and nitrate in subsurface wastewater infiltration system: Implication for reducing greenhouse gas emissions.

Subsurface wastewater infiltration system (SWIS) has been recognized as a cost-effective and environmentally friendly tool for wastewater treatment. However, there is a lack of knowledge on the transformation processes of nitrogen (N), hindering the improvement of the N removal efficiency in SWIS. Here, the migration and transformation mechanisms of ammonium (NH4+-N) and nitrate (NO3+-N) over 10 days were explored by 15N labeling technique. Over the study period, 49% of the added 15NH4+-N remained in the soil, 29% was removed via gaseous N emissions, and 14% was leaked with the effluent in the SWIS. In contrast, only 11% of the added 15NO3--N remained in the soil while 65% of the added 15NO3--N was removed via gaseous N emissions, and 12% with the effluent in the SWIS. The main pathway for N2O emission was denitrification (52-70%) followed by nitrification (15-28%) and co-denitrification (9-20%). Denitrification was also the predominant pathway for N loss as N2, accounting for 88-96% of the N2 emission. The dominant biological transformation processes were different at divergent soil depths, corresponding to nitrification zone and denitrification zone along the longitudinal continuum in SWIS, which was confirmed by the expression patterns of microbial gene abundance. Overall, our findings reveal the mechanism of N transformation in SWIS and provide a theoretical basis for establishing a pollutant management strategy and reducing greenhouse gas emissions from domestic wastewater treatment.

Microbial-Related Metabolites May Be Involved in Eight Major Biological Processes and Represent Potential Diagnostic Markers in Gastric Cancer.

Metabolites associated with microbes regulate human immunity, inhibit bacterial colonization, and promote pathogenicity. Integrating microbe and metabolome research in GC provides a direction for understanding the microbe-associated pathophysiological process of metabolic changes and disease occurrence. The present study included 30 GC patients with 30 cancerous tissues and paired non-cancerous tissues (NCs) as controls. LC-MS/MS metabolomics and 16S rRNA sequencing were performed to obtain the metabolic and microbial characteristics. Integrated analysis of the microbes and metabolomes was conducted to explore the coexistence relationship between the microbial and metabolic characteristics of GC and to identify microbial-related metabolite diagnostic markers. The metabolic analysis showed that the overall metabolite distribution differed between the GC tissues and the NC tissues: 25 metabolites were enriched in the NC tissues and 42 metabolites were enriched in the GC tissues. The α and ß microbial diversities were higher in the GC tissues than in the NC tissues, with 11 differential phyla and 52 differential genera. In the correlation and coexistence integrated analysis, 66 differential metabolites were correlated and coexisted, with specific differential microbes. The microbes in the GC tissue likely regulated eight metabolic pathways. In the efficacy evaluation of the microbial-related differential metabolites in the diagnosis of GC, 12 differential metabolites (area under the curve [AUC] >0.9) exerted relatively high diagnostic efficiency, and the combined diagnostic efficacy of 5 to 6 microbial-related differential metabolites was higher than the diagnostic efficacy of a single feature. Therefore, microbial diversity and metabolite distribution differed between the GC tissues and the NC tissues. Microbial-related metabolites may be involved in eight major metabolism-based biological processes in GC and represent potential diagnostic markers.

Expression characteristics and their functional role of IGFBP gene family in pan-cancer.

BACKGROUND: Insulin-like growth factor binding proteins (IGFBPs) are critical regulators of the biological activities of insulin-like growth factors. The IGFBP family plays diverse roles in different types of cancer, which we still lack comprehensive and pleiotropic understandings so far. METHODS: Multi-source and multi-dimensional data, extracted from The Cancer Genome Atlas (TCGA), Oncomine, Cancer Cell Line Encyclopedia (CCLE), and the Human Protein Atlas (HPA) was used for bioinformatics analysis by R language. Immunohistochemistry and qRT-PCR were performed to validate the results of the database analysis results. Bibliometrics and literature review were used for summarizing the research progress of IGFBPs in the field of tumor. RESULTS: The members of IGFBP gene family are differentially expressed in various cancer types. IGFBPs expression can affect prognosis of different cancers. The expression of IGFBPs expression is associated with multiple signal transduction pathways. The expression of IGFBPs is significantly correlated with tumor mutational burden, microsatellite instability, tumor stemness and tumor immune microenvironment. The qRT-PCR experiments verified the lower expression of IGFBP2 and IGFBP6 in gastric cancer and the lower expression of IGFBP6 in colorectal cancer. Immunohistochemistry validated a marked downregulation of IGFBP2 protein in gastric cancer tissues. The keywords co-occurrence analysis of IGFBP related publications in cancer showed relative research have been more concentrating on the potential of IGFBPs as tumor diagnostic and prognostic markers and developing cancer therapies. CONCLUSIONS: These findings provide frontier trend of IGFBPs related research and new clues for identifying novel therapeutic targets for various cancers.

Effect of increased carbon load on denitrification efficiency and nitrate isotope enrichment factors in subsurface wastewater infiltration system.

Denitrification plays a dominant role in nitrate removal in subsurface wastewater infiltration system (SWIS). However, the effect of increased carbon (C) load on denitrification efficiency in the SWIS remain unclear. In this study, we used analyses of stable isotopes of nitrogen (N) and oxygen (O) in nitrate to investigate the N and O isotope enrichment factors (15 ε and 18 ε) and quantified N losses via denitrification in SWIS. The results demonstrated that an increase in C loads positively affected the pollutant removal performance of SWIS. The natural abundance of 15 N and 18 O increased with decreasing nitrate concentration from 12.5 to 7.3 mg/L, accompanied by increased 15 ε and 18 ε from -8.7‰ to -10.6‰ and -5.9‰ to -8.2‰, respectively, as the C load increased from 18 to 36 g/(m2  d). The contribution of denitrification to nitrate removal was 62%, 71%, and 77% when C loads were 18, 27, and 36 g/(m2  d), respectively, indicating that increased C loads could improve the nitrate removal through denitrification in SWIS. PRACTITIONER POINTS: Increasing C loads positively affected the nitrate removal performance of SWIS. N and O isotope enrichment factors of nitrate increased with the enhancement of influent C load. A C load of 36 g/(m2 d) is recommended in SWIS to improve the N removal performance and denitrification efficiency.

Ultrasensitive determination of α-glucosidase activity using CoOOH nanozymes and its application to inhibitor screening.

In this work, a novel method for the colorimetric sensing of α-glucosidase (α-Glu) activity was developed based on CoOOH nanoflakes (NFs), which exhibit efficient oxidase-mimicking activity. Colorless 3,3',5,5'-tetramethylbenzidine (TMB) can be oxidized by CoOOH NFs into blue-colored oxidized TMB (oxTMB) in the absence of H2O2. L-Ascorbic acid-2-O-α-D-glucopyranose (AAG) can be hydrolysed by α-glucosidase to produce ascorbic acid, resulting in a significant decrease of catalytic activity of CoOOH NFs. Thus, a colorimetric α-glucosidase activity detection method was designed with a limit of detection of 0.0048 U mL-1. Furthermore, the designed sensing platform exhibits favorable applicability for the α-glucosidase (α-Glu) activity assay in real samples. Meanwhile, this method can be expanded to study the inhibitors of α-Glu. Finally, the as-proposed method combined with a smartphone would be a color recognizer, which was successfully applied for the determination of α-Glu activity in human serum samples.

Numerical Studies on Failure Mechanisms of All-Composite Sandwich Structure with Honeycomb Core under Compression and Impact Loading Conditions.

The all-composite sandwich structure with the honeycomb core is a lightweight and high-strength structure with broad application scenarios. The face sheet and honeycomb core of the proposed all-composite sandwich structure in this work are composed of carbon-fiber-reinforced polymer (CFRP) composites. The mechanical response and damage mechanism of the all-composite sandwich structure under out-of-plane quasi-static compression and out-of-plane impact are studied by numerical methods. The refined finite element models of the sandwich structures are built on the ABAQUS/Explicit platform. The micromechanics of failure (MMF) theory based on physical component failure is used to describe the intralaminar damage mechanism of the face sheet and honeycomb core, and the mixed-mode exponential cohesive zone model (ECZM) is utilized to simulate the initiation and evolution of interlayer damage. In addition, the cohesive contact approach is adopted to capture the debonding failure at the face-sheet/core. The numerical results show that the all-composite sandwich structure has the characteristics of large structural stiffness and strong energy absorption ability. The failure mechanism of the all-composite sandwich structure under compression is mainly matrix damage and delamination of the honeycomb core, with buckling and folding in appearance. Under out-of-plane impact, matrix damage and delamination arise on the upper sheet, little damage is observed on the lower sheet, and the delamination damage morphology tends to be circular with increasing impact energy. In addition, the interface failure of the upper-sheet/core is more than that of the lower-sheet/core. In addition, the matrix damage near the impact center of the honeycomb core tends to be consistent with the delamination contour, and a small amount of fiber failure is also observed, which manifests as a collapse morphology of the impact area. The research results enrich the understanding of the mechanical behavior of all-composite sandwich structures with honeycomb cores and provide theoretical support for their potential applications.

Comprehensive Analysis and Validation of Solute Carrier Family 25 (SLC25) and Its Correlation with Immune Infiltration in Pan-Cancer.

As the largest gene family functioning in protein transport among human solute carriers, the SLC25 family (mitochondrial carrier family) can participate in development of cancer. However, a comprehensive exploration for the exactly roles of SLC family remains lacking. In the present study, a total of 15 functional SLC25 family genes were retrieved from all current publications. And multidimensional analyses were systematically performed based on the transcriptome and genome data of SLC25 family from a variety of online databases for their expression, immune cell infiltration, and cancer prognosis. Validation by qPCR and immunohistochemistry were further conducted for the expression of partial SLC25 family members in some tumor tissue. We found that the SLC25 family had strong correlation with immune cells, such as macrophages M2, CD8+ T cell, CD4+ T cell memory activated, and memory resting. Among them, SLC25A6 was most correlated with Macrophage M1 in uveal melanoma (r = -0.68, P = 1.9e - 0.5). Expression of mRNA level showed that SLC25A4 was downregulated in stomach adenocarcinoma and colon adenocarcinoma. SLC25A7 was highly expressed in stomach adenocarcinoma and colon adenocarcinoma. SLC25A23 was decreased in colon adenocarcinoma. qPCR and immunohistochemistry validation results were consistent with our bioinformatics prediction. SLC25A8 was associated with the prognosis of cancer. All these findings suggested that the SLC25 family might affects the immune microenvironment of the cancer and then had the potential to be predictive biomarkers for early diagnosis and prognosis as well as novel targets for individualized treatment of cancer.

Epidemiology of tick-borne pathogens of cattle and tick control practices in coastal Kenya.

Tick-borne diseases (TBD) are a major constraint to livestock health and productivity in sub-Saharan Africa. Nonetheless, there are relatively few robust epidemiologic studies documenting TBD and its management in different endemic settings in Kenya. Therefore, a cross-sectional study using multi-stage cluster sampling was undertaken to characterize the epidemiology of TBD and management factors among zebu cattle reared under an extensive system in coastal Kenya. Blood samples from 1486 cattle from 160 herds in 14 villages were screened for the presence of tick-borne bacterial and protozoan pathogens using PCR with high-resolution melting analysis and sequencing. Standardized questionnaires were used to collect data on herd structure and herd management practices, and a mixed-effect logistic regression model to identify risk factors for tick-borne pathogens (TBPs). The application of chemical acaricide was the primary method for tick control (96.3%, 154/160), with the amidine group (mainly Triatix®, amitraz) being the most frequently used acaricides. Respondents identified East Coast fever as the most important disease and Butalex® (buparvaquone) was the most commonly administered drug in response to perceived TBD in cattle. The overall animal- and herd-level prevalence for TBPs were 24.2% (95% confidence interval (CI): 22.0-26.4%) and 75.6% (95% CI: 68.2-82.1%), respectively. Cattle were infected with Anaplasma marginale (10.9%, 95% CI: 9.4-12.6), Theileria parva (9.0%, 95% CI: 7.5-10.5), Anaplasma platys (2.6%, 95% CI: 1.9-3.6), Theileria velifera (1.1%, 95% CI: 0.7-1.8), Babesia bigemina (0.5%, 95% CI: 0.2-1.0), and Anaplasma sp. (0.1%, 95% CI: 0.0-0.4). Moreover, 21 cattle (1.4%) were co-infected with two TBPs. None of the assessed potential risk factors for the occurrence of either A. marginale or T. parva in cattle were statistically significant. The intra-herd correlation coefficients (lCCs) computed in this study were 0.29 (A. marginale) and 0.14 (T. parva). This study provides updated molecular-based information on the epidemiological status of TBPs of cattle and herd management practices in coastal Kenya. This information can be used in designing cost-effective control strategies for combating these TBD in the region.

RNA-sequencing reveals the expression profiles of tsRNAs and their potential carcinogenic role in cholangiocarcinoma.

BACKGROUND: Recently, the incidence of cholangiocarcinoma (CCA) has gradually increased. As CCA has a poor prognosis, the ideal survival rate is scarce for patients. The abnormal expressed tsRNAs may regulate the progression of a variety of tumors, and tsRNAs is expected to become a new diagnostic biomarker of cancer. However, the expression of tsRNAs is obscure and should be elucidated in CCA. METHODS: High-throughput RNA sequencing technology (RNA-seq) was utilized to determine the overall expression profiles of tsRNAs in three pairs CCA and adjacent normal tissues and to screen the tsRNAs that were differentially expressed. The target genes of dysregulated tsRNAs were predicted and the biological effects and potential signaling pathways of these target genes were explored by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to validate 11 differentially expressed tRFs with 12 pairs CCA and adjacent normal tissues. RESULTS: High-throughput RNA-seq totally demonstrated 535 dysregulated tsRNAs, of which 241 tsRNAs were upregulated, such as tRF-21-YLKZKWE5D，tRF-16-9NF5W8B，tRF-27-78YLKZKWE52，tRF-19-RLXN48KP，tRF-33-IK9NJ4S2I7L7DV，tRF-19-F8DHXYIV, and 294 tsRNAs were downregulated (tRF-20-739P8WQ0, tRF-34-JJ6RRNLIK898HR, tRF-17-VL8RPY5, tRF-23-YP9LON4VDP, tRF-39-EH623K76IR3DR2I2, tRF-17-18YKISM, tRF-19-Q1Q89PJZ, etc.) in CCA compared with adjacent normal tissues (|log2 [fold change] | ≥ 1 and p value <0.05). GO and KEGG enrichment analyses indicated that the target genes of dysregulated tRFs (tRF-34-JJ6RRNLIK898HR, tRF-38-0668K87SERM492V, and tRF-39-0668K87SERM492E2) were mainly enriched in the Notch signaling pathway, Hippo signaling pathway, cAMP signaling pathway and in growth hormone synthesis, secretion and action, etc. qRT-PCR result showed that tRF-34-JJ6RRNLIK898HR/tRF-38-0668K87SERM492V/tRF-39-0668K87SERM492E2 was downregulated (p = 0.021), and tRF-20-LE2WMK81 was upregulated in CCA (p = 0.033). CONCLUSION: Differentially expressed tRFs in CCA are enriched in many pathways associated with neoplasms, which may impact the tumor progression and have potential to be diagnostic biomarkers and therapeutic targets of CCA.

Evaluating the efficacy of Mazao Tickoff (Metarhizium anisopliae ICIPE 7) in controlling natural tick infestations on cattle in coastal Kenya: Study protocol for a randomized controlled trial.

Ticks and tick-borne diseases cause substantial economic losses to the livestock industry in sub-Saharan Africa. Mazao Tickoff is a novel bioacaricide developed for tick control and is based on the entomopathogenic fungus Metarhizium anisopliae sensu lato (s.l.) isolate ICIPE 7. To date, no randomized controlled study has been undertaken to demonstrate the efficacy of this bioacaricide in reducing natural tick infestation on cattle. To this end, this field trial is designed to evaluate the anti-tick efficacy of Mazao Tickoff on cattle in coastal Kenya compared to a standard chemical tick control protocol. In this prospective, multi-center randomized controlled trial, eligible herds will be randomized by the herd size to the intervention arm in a 1:1:1 ratio to either Triatix® (active ingredient: amitraz); Mazao Tickoff (active ingredient: M. anisopliae ICIPE 7); or placebo (excipients of the Mazao Tickoff), with a total enrollment target of 1,077 cattle. Treatments will be dispensed on Day 0 (defined individually as the day each animal receives the first treatment) and thereafter every two weeks until Day 182. Ticks will be counted on every animal in each herd (herds to be included have at least one animal bearing at least one tick on Day 0), and thereafter on bi-weekly intervals until Day 182. The primary efficacy assessments of Mazao Tickoff will be based on the mean percentage reduction in tick counts at each post-treatment follow-up visit compared to the placebo group and the Triatix® arm. Further, the effect of Mazao Tickoff on the prevalence of common cattle pathogens, Anaplasma marginale and Theileria parva, will be determined by assessing incidence and seroprevalence at four different time points. This protocol describes the first rigorous evaluation of the efficacy of Mazao Tickoff and its potential as a viable alternative non-chemical acaricide tool for tick control in Kenya and elsewhere.

Retention of deposited ammonium and nitrate and its impact on the global forest carbon sink.

The impacts of enhanced nitrogen (N) deposition on the global forest carbon (C) sink and other ecosystem services may depend on whether N is deposited in reduced (mainly as ammonium) or oxidized forms (mainly as nitrate) and the subsequent fate of each. However, the fates of the two key reactive N forms and their contributions to forest C sinks are unclear. Here, we analyze results from 13 ecosystem-scale paired 15N-labelling experiments in temperate, subtropical, and tropical forests. Results show that total ecosystem N retention is similar for ammonium and nitrate, but plants take up more labelled nitrate ([Formula: see text]%) ([Formula: see text]) than ammonium ([Formula: see text]%) while soils retain more ammonium ([Formula: see text]%) than nitrate ([Formula: see text]%). We estimate that the N deposition-induced C sink in forests in the 2010s  is [Formula: see text] Pg C yr-1, higher than previous estimates because of a larger role for oxidized N and greater rates of global N deposition.

Interleukin 20 receptor A expression in colorectal cancer and its clinical significance.

BACKGROUND: Interleukin 20 receptor A (IL20RA) has been shown to play a role in the establishment and progression of multiple tumors. However, the expression of this protein in colorectal cancer (CRC) and its correlation with the clinicopathological parameters of CRC have remained unclear. METHODS: A total of 323 paraffin sections including CRC tissues and adjacent normal tissues after surgery were collected. IL20RA protein expression was detected by immunohistochemical staining. The difference expression of IL20RA mRNA between CRC and normal tissues was also explored in the Oncomine and GEO databases. In addition, the IL20RA-related differentially expressed genes were analyzed in TCGA database and enrichment analysis was conducted to explore the cell functions and pathways related to IL20RA expression. RESULTS: There was increased IL20RA expression in CRC compared with that in normal tissues. High IL20RA expression was associated with greater tumor diameter, lymph node metastasis, and poor TNM stage in CRC, while also being suggestive of poor prognosis. The main pathways of IL20RA-related differentially expressed genes in TCGA were protein heterodimerization activity, oxygen binding, oxygen transporter activity, hormone activity, and lipid transporter activity. Meanwhile, IL20RA-related differentially expressed genes were mainly enriched in peroxidase, nucleotide stimulant repair, fatty acid metabolism, basal transcription factor, and RNA degradation. CONCLUSIONS: IL20RA might have a role as a biomarker for CRC. Its upregulation might contribute to an aggressive phenotype in CRC. IL20RA's involvement in the development and progression of CRC might occur through it affecting fatty acid metabolism, oxygen binding, oxygen transport, and hormone activity.

Comprehensive Pan-Cancer Analysis of Heat Shock Protein 110, 90, 70, and 60 Families.

Background: Here we carried out a panoramic analysis of the expression and prognosis of HSP110, HSP90, HSP70, and HSP60 families in 33 types of cancer, with the aim of deepening the systematic understanding of heat shock proteins (HSPs) in cancer. Materials and Methods: Next-generation sequencing data of multiple tumors were downloaded from TCGA, CCLE and Oncomine databases. RStudio 3.6.1 was used to analyze HSP110, HSP90, HSP70 and HSP60 families based on their expression in 33 types of cancer. The validations in vivo (stomach adenocarcinoma and colon adenocarcinoma tissues) were performed by qRT-PCR. Results: HSPs were differentially expressed in different cancers. The results revealed mainly positive correlations among the expressions of HSPs in different cancers. Expressions of HSP family members were generally associated with poor prognosis in respiratory, digestive, urinary and reproductive system tumors and associated with good prognosis in cholangiocarcinoma, pheochromocytoma and paraganglioma. TCGA mutation analysis showed that HSP gene mutation rate in cancers was 0-23%. CCLE mutation analysis indicated that HSP gene mutation rate in 828 cell lines from 15 tumors was 0-17%. CNV analysis revealed that HSPs have different degrees of gene amplifications and deletions in cancers. Gene mutations of 15 HSPs influenced their protein expressions in different cancers. Copy number amplifications and deletions of 22 HSPs also impacted protein expression levels in pan-cancer. HSP gene mutation was generally a poor prognosis factor in cancers, except for uterine corpus endometrial carcinoma. CNVs in 14 HSPs showed varying influences on survival status in different cancers. HSPs may be involved in the activation and inhibition of multiple cancer-related pathways. HSP expressions were closely correlated with 22 immune cell infiltrations in different cancers. The qRT-PCR validation results in vivo showed that HSPA2 was down-regulated in stomach adenocarcinoma and colon adenocarcinoma; HSPA7 and HSPA1A also were down-regulated in colon adenocarcinoma. HSPA2-HSPA7 (r = 0.031, p = 0.009) and HSPA1A-HSPA7 (r = 0.516, p < 0.001) were positive correlation in colon adenocarcinoma. Conclusion: These analysis and validation results show that HSP families play an important role in the occurrence and development of various tumors and are potential tumor diagnostic and prognostic biomarkers as well as anti-cancer therapeutic targets.

Quantifying nitrogen uptake and translocation for mature trees: an in situ whole-tree paired 15N labeling method.

Nitrogen (N) is one of the major nutrients limiting plant growth in terrestrial ecosystems. To avoid plant-microbe competition, previous studies on plant N uptake preference often used hydroponic experiments on fine roots of seedlings and demonstrated ammonium preference for conifer species; however, we lack information about N uptake and translocation in the field. In this paper, we described a method of in situ paired 15N labeling and reported the rates and time course of N uptake and translocation by mature trees in situ. We added 15N-enriched ammonium or nitrate, together with the nitrification inhibitor dicyandiamide, to paired Larix kaempferi (Lamb.) Carr (larch) trees from 30-, 40- and 50-year-old plantations. Fine roots, coarse roots, leaves and small branches were collected 2, 4, 7, 14 and 30 days after labeling. Nitrate uptake and translocation averaged 1.59 ± 0.16 µg 15N g-1 day-1, which is slightly higher than ammonium (1.08 ± 0.10 µg 15N g-1 day-1), in all tree organs. Nitrate contributed 50-78% to N uptake and translocation, indicating efficient nitrate use by larch in situ. We observed no age effect. We suggest that sampling leaves after 4 days of 15N labeling is sufficient to detect mature tree N uptake preference in situ. Whole-tree 15N-ammonium recovery equaled that of 15N-nitrate 30 days after 15N addition, implying the importance of both ammonium and nitrate to mature larch N use in the long run. We conclude that our method is promising for studying mature tree N uptake preference in situ and can be applied to other conifer and broadleaf species. We suggest using highly enriched 15N tracer to overcome soil dilution and a nitrification inhibitor to minimize ammonium transformation to nitrate. Our study revealed mature tree N preference in situ and demonstrated the strong contribution of nitrate toward mature larch growth on soils rich in nitrate.

Dynamics and multi-annual fate of atmospherically deposited nitrogen in montane tropical forests.

The effects of nitrogen (N) deposition on forests largely depend on its fate after entering the ecosystem. While several studies have addressed the forest fate of N deposition using 15 N tracers, the long-term fate and redistribution of deposited N in tropical forests remains unknown. Here, we applied 15 N tracers to examine the fates of deposited ammonium ( NH 4 + ) and nitrate ( NO 3 - ) separately over 3 years in a primary and a secondary tropical montane forest in southern China. Three months after 15 N tracer addition, over 60% of 15 N was retained in the forests studied. Total ecosystem retention did not change over the study period, but between 3 months and 3 years following deposition 15 N recovery in plants increased from 10% to 19% and 13% to 22% in the primary and secondary forests, respectively, while 15 N recovery in the organic soil declined from 16% to 2% and 9% to 2%. Mineral soil retained 50% and 35% of 15 N in the primary and secondary forests, with retention being stable over time. The total ecosystem retention of the two N forms did not differ significantly, but plants retained more 15 NO 3 - than 15 NH 4 + and the organic soil more 15 NH 4 + than NO 3 - . Mineral soil did not differ in 15 NH 4 + and 15 NO 3 - retention. Compared to temperate forests, proportionally more 15 N was distributed to mineral soil and plants in these tropical forests. Overall, our results suggest that atmospherically deposited NH 4 + and NO 3 - is rapidly lost in the short term (months) but thereafter securely retained within the ecosystem, with retained N becoming redistributed to plants and mineral soil from the organic soil. This long-term N retention may benefit tropical montane forest growth and enhance ecosystem carbon sequestration.

Comparison of clinicopathological parameters, prognosis, micro-ecological environment and metabolic function of Gastric Cancer with or without Fusobacterium sp. Infection.

Background: Fusobacterium sp. plays a crucial role in the tumorigenesis and development of gastrointestinal tumors. Our research group previously disclosed that Fusobacterium sp. was more abundant in gastric cancer (GC) tissues than adjacent non-cancerous (NC) tissues. However, Fusobacterium sp. did not exist in all GC tissues and the differentiated features of GC with or without Fusobacterium sp. infection is not clear. Methods: The expression data of 61 GC tissues came from 16S rRNA gene sequencing. Comparison groups were defined based on sOTU at the genus level of Fusobacterium sp., which was performed by the Qiime2 microbiome bioinformatics platform. We used Chi-square and Fisher's exact test to compare clinicopathological parameters, and used Kaplan-Meier analysis, Cox univariate and multivariate analysis to compare prognosis. Micro-ecological environment comparison was characterized by 16S rRNA gene sequencing, and the metabolic function prediction was applied by PICRUSt2. Results of microbial diversity, differential enrichment genus and metabolic function in GC with or without Fusobacterium sp. infection was validated with 229 GC tissues downloaded from an independent cohort in ENA database (PRJNA428883). Results: The infection rate of Fusobacterium sp. in 61 GC tissues was 52.46% and elderly GC patients were more prone to Fusobacterium sp. infection. GC patients infected with Fusobacterium sp. were more likely to have tumor-infiltrating lymphocytes and p53 expression. The microbial diversity and microbial structure showed significant differences between two GC tissue groups with 42 differential enrichment genera. The metabolic function of Fusobacterium sp.-positive GC tissues was related to the biosynthesis of lysine, peptidoglycan, and tRNA. The differences in microbial structure, the existence of some differential enrichment genera and the metabolic function of Fusobacterium sp.-positive GC tissues, were then validated by 229 GC tissues of an independent cohort. Conclusions: Fusobacterium sp. infection can affect the phenotypic characteristics, micro-ecological environment, and metabolic functions of GC, which may provide a basis for further exploring the relationship between Fusobacterium sp. infection and carcinogenesis of GC.

Mitochondria-related core genes and TF-miRNA-hub mrDEGs network in breast cancer.

BACKGROUND: Mitochondria-nuclear cross-talk and mitochondrial retrograde regulation are involved in the genesis and development of breast cancer (BC). Therefore, mitochondria can be regarded as a promising target for BC therapeutic strategies. The present study aimed to construct regulatory network and seek the potential biomarkers of BC diagnosis and prognosis as well as the molecular therapeutic targets from the perspective of mitochondrial dysfunction. METHODS: The microarray data of mitochondria-related encoding genes in BC cell lines were downloaded from GEO including GSE128610 and GSE72319. GSE128610 was treated as test set and validation sets consisted of GSE72319 and TCGA tissue samples, intending to identify mitochondria-related differentially expressed genes (mrDEGs). We performed enrichment analysis, PPI network, hub mrDEGs and overall survival analysis and constructed transcription factor (TF)-miRNA-hub mrDEGs network. RESULTS: A total of 23 up-regulated and 71 down-regulated mrDEGs were identified and validated in BC cell lines and tissues. Enrichment analyses indicated that mrDEGs were associated with several cancer-related biological processes. Moreover, 9 hub mrDEGs were identified and validated in BC cell lines and tissues. Finally, 5 hub coregulated mrDEGs, 21 miRNAs and 117 TFs were used to construct TF-miRNA-hub mrDEGs network. MYC associated zinc finger protein (MAZ), heparin binding growth factor (HDGF) and Sp2 transcription factor (SP2) regulated 3 hub mrDEGs. Hsa-mir-21-5p, hsa-mir-1-3p, hsa-mir-218-5p, hsa-mir-26a-5p and hsa-mir-335-5p regulated 2 hub mrDEGs. Overall survival analysis suggested that the up-regulation of fibronectin 1 (FN1), as well as the down-regulation of discoidin domain receptor tyrosine kinase 2 (DDR2) correlated with unfavorable prognosis in BC. CONCLUSION: TF-miRNA-hub mrDEGs had instruction significance for the exploration of BC etiology. The hub mrDEGs such as FN1 and DDR2 were likely to regulate mitochondrial function and be novel biomarkers for BC diagnosis and prognosis as well as the therapeutic targets.