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Heterogeneous karst surfaces exerted scaling effects whereby specific runoff decrease with increasing area. The existing RUSLE-L equations are limited by the default implicit assumption that the surface-runoff intensity is constant at any slope length. The objective of this study was to modify the L-equation by establishing the functional relationship between surface-runoff intensity and karst slope length, and to evaluate its predictive capability at different resolution DEMs. Transfer grid layers were generated based on the area rate of surface karstification and considered the runoff transmission percentage at the exposed karst fractures or conduits to be zero. Using the multiple flow direction algorithm united with the transfer grid (MFDTG), the flow accumulation of each grid cell was simulated to estimate the average surface-runoff intensity over different slope lengths. The effectiveness of MFDTG algorithm was validated by runoff plot data in Southwestern China. The simulated results in a typical peak-cluster depression basin with an area rate of surface karstification of 6.5% showed that the relationship between surface-runoff intensity and slope length was a negative power function. Estimated by the proposed modified L-equation ((alx(b+1)/22.13)m), the L-factor averages of the study basin ranged from 0.35 to 0.41 at 1, 5, 25 and 90 m resolutions respectively. This study indicated that the modified L-equation enables an improved prediction of the much smaller L-factor and the use of any resolution DEMs on karst landscapes. Particular attention should be given to the variation of surface-runoff intensity with slope length when predicting L-factor on hillslopes with runoff scale effect.

Objective The thickness of soilepikarst is an important index affecting the hydrological process of critical zones. The spatial heterogeneity is critical understanding the structural evolution mechanism of the key zones of the Earth and the water conservation function of the key zones of the Earth. Methods Based on the 1.4 km2 area of the Mulian catchment in Huanjiang, Guangxi, the 1 731 soil-epikarst thickness samples were obtained through electrical resistivity tomography (ERT) with 45 sampling lines.The spatial distribution pattern of soil and epikarst thickness and its influencing factors were investigated. The results showed that the average thickness of soil and epikarst was 1.15 m and 6.44 m with strong and medium variance, respectively. Results The results of geostatistical analysis indicated that the spherical model and exponential model could reflect the spatial structure characteristics of soil and epikarst thickness, respectively. Soil thickness showed moderate spatial autocorrelation, a long range and good spatial continuity. However, the epikarst presented strong spatial autocorrelation and spatial dependence with a short range. Soil thickness was affected by multiple environmental factors (topographic wetness index, vertical curvature, curvature, aspect, slope, elevation, coverage, outcrop ratio and NDVI), while the thickness of epikarst was more affected by soil thickness and vegetation type. Conclusion These results are helpful for understanding the evolution of the soil-epikarst zone and provide a scientific basis for the spatial prediction of soil-epikarst thickness in karst areas.

On karst steep slopes, subsurface flow frequently emerges at the rock-soil interface, thereby exacerbating soil erosion. However, there have been few studies that quantitatively assess the impact of subsurface flow on soil erosion in this region. We conducted indoor artificial rainfall experiments to observe runoff and sediment yield under two conditions: with subsurface flow (SF) and without subsurface flow input (WSF). We considered various rainfall intensities (30 mm h−1, 60 mm h−1, 90 mm h−1) and slope gradients (30°, 35°, 40°). The results demonstrated that the presence of subsurface flow significantly amplified both surface runoff and sediment yield, while also intensifying the influence of rainfall intensity and slope gradient on runoff and sediment yield. Compared to WSF, under SF conditions, surface runoff increased by 1.55 to 4.43 times, and sediment yield increased by 4.60 to 8.72 times. Slope gradient played a substantial role in soil loss, while rainfall intensity primarily impacted surface runoff. Under WSF conditions, soil loss only occurred at a 40° slope gradient or under a rainfall intensity of 90 mm h−1, whereas under SF conditions, soil loss could occur at a 30° slope gradient or under a rainfall intensity of 30 mm h−1. This research provides a scientific foundation for a more profound understanding of the mechanisms driving soil erosion on karst slopes, particularly in the context of subsurface flow.

Urtica cannabina L. (UL) has been used clinically for centuries because of its anti-inflammatory properties. This study aimed to investigate the underlying mechanisms and anti-inflammatory effects of different UL concentrations in zebrafish. To elucidate UL's anti-inflammatory properties, two inflammation zebrafish models were designed 1) by severing the zebrafish's caudal fin to assess the repairing effect of UL on the tail inflammation, and 2) by inducing lipopolysaccharides (LPS)-mediated intestinal inflammation to assess the protective and reparative effects of UL on intestinal inflammation at the histological and genetic levels. Furthermore, the effect of UL on the LPS-induced intestinal flora changes was also assessed. After caudal fin resection, a scar formed on the tail of the zebrafish, and the area of the caudal fin increased by 1.30 times as much as that of the control group (P < 0.01). Moreover, this tail scar was alleviated after 10 mg/g UL supplementation but not after 30 mg/g UL dose. LPS decreased the feed intake and body weight of the zebrafish; however, these effects were reversed after 10 and 30 mg/g doses of UL. In addition, the LPS treatment also reduced the intestinal goblet cells by 49% in the zebrafish when compared with the control, which was significantly restored after 10 and 30 mg/g UL treatments. At the genetics level, the expression of the pro-inflammatory cytokine genes (TNF-α, IL6, and IL8) showed that 10 and 30 mg/g UL doses could rescue LPS-induced expression. The gut microbiota analysis revealed changes in the abundance of four major bacterial phyla in the 10 and 30 mg/g UL-treated groups, with an increased probiotic Bacteroidota and decreased pathogenic bacteria. These results indicate that UL strongly inhibits inflammation caused by caudal fin removal and LPS-induced inflammatory changes in the zebrafish intensity, suggesting that UL is a feed additive that could be developed to improve resistance to inflammation in livestock.

Soils provide storage space for nutrients and water, which governs many hydrological processes and functions in the natural ecosystems. Due to the complex soil-rock structures and localized climate, soil thickness and stock vary distinctly in different karst areas in southwest China. The difficulties of soil detection and the non-transparent underground soil-rock structures limit a deeper understanding of the spatial distribution of soils and their ecological functions. In this study, we employed electrical resistivity tomography (ERT) combined with soil probes to detect the spatial distribution of soils on hillslopes in karst forests in southwest China. The ERT surveys were conducted at six sites with 19 plots (20 × 30 m2) along latitudinal gradients. The results show that the ERT method performs well in visualizing the complex soil–bedrock interfaces and patchy distribution of soils in karst hillslopes, with remarkable consistency with the observed soil thickness and rock outcrops. Soil probes often hit rocks in soils (which we call a “rock barrier effect”), resulting in an underestimation of the mean soil thickness within the one-meter exploration depth by ∼0.18 m. This indicates that soil stock estimates derived from soil probes may be obviously underestimated in karst areas where the rock barrier effects are widespread. Meanwhile, the concept of equivalent soil thickness is proposed to remove the impact of scattered bedrock on soil stock estimation. For the six selected sites, the ERT-inferred mean soil thickness, equivalent soil thickness, and stock are 0.86 m, 0.60 m, and 0.10 m3/m3 respectively, with a trend of decreasing from high to low latitudes. Moreover, the rock outcrop coverage rate explains 52 % (60 %) of the variation in soil thickness (stock), followed by the annual mean temperature. This study emphasizes the importance of considering bedrock topography and its distribution in improving the detection of soil distribution, and the implications of the findings in this study are perhaps most profound for soil thickness-related soil erosion, soil water storage, and carbon storage in karst critical zones.

A series of novel quinoline-O-carbamate derivatives was rationally designed for treating Alzheimer’s disease (AD) by multi-target-directed ligands (MTDLs) strategy. The target compounds were synthesised and evaluated by AChE/BuChE inhibition and anti-inflammatory property. The in vitro activities showed that compound 3f was a reversible dual eeAChE/eqBuChE inhibitor with IC50 values of 1.3 µM and 0.81 µM, respectively. Moreover, compound 3f displayed good anti-inflammatory property by decreasing the production of IL-6, IL-1β and NO. In addition, compound 3f presented significant neuroprotective effect on Aβ25-35-induced PC12 cell injury. Furthermore, compound 3f presented good stabilities in artificial gastrointestinal fluids, liver microsomes in vitro and plasma. Furthermore, compound 3f could improve AlCl3-induced zebrafish AD model by increasing the level of ACh. Therefore, compound 3f was a promising multifunctional agent for the treatment of AD.

Abstract Background and Aim Programmed death 1 (PD‐1) and programmed death ligand 1 (PD‐L1) inhibitors have transformed the treatment landscape of advanced hepatocellular carcinoma (HCC), but consistent responses are not observed in all patients, and prognostic biomarkers to guide treatment decisions are lacking. We aimed to evaluate the predictive value of PD‐L1 expression in advanced HCC patients treated with PD‐1/PD‐L1 inhibitors. Methods A comprehensive search of PubMed, Embase, Web of Science, and the Cochrane Library was conducted. Studies comparing the objective response rate (ORR) and/or disease control rate (DCR) based on the tumor PD‐L1 status of HCC were included. Results Eleven studies with 1,330 HCC patients treated with PD‐1/PD‐L1 inhibitors were included. Pooled odds ratio (OR) analysis demonstrated a significantly improved ORR in PD‐L1‐positive patients compared with PD‐L1‐negative patients (OR, 1.86, 95% CI, 1.35–2.55). Similar results were observed in the anti‐PD‐1 treatment (p

Abstract Introduction Due to the high recurrence, the HCC prognosis remains poor. Yet, the biomarkers for predicting the recurrence of high‐risk patients are currently lacking. We aimed to develop a signature to predict the recurrence of HCC based on NKG2D ligands. Methods The multivariate Cox proportional hazards regression was used to select recurrence‐related variables of NKG2D ligands in HCC patients from The Cancer Genome Atlas (TCGA). HCC patients from the OEP000321 dataset and Guilin cohort were used to validate the predictive signature. The mRNA expression of NKG2D ligands was measured by QRT‐PCR. Immunohistochemistry analysis of HCC tissue microarray samples was used to identify the expression of NKG2D ligands. Results In this study, NKG2D ligands expression in the mRNA and protein level was both abnormally expressed in HCC and associated with recurrence‐free survival (RFS). Then, the recurrence‐related variables of NKG2D ligands in HCC were selected by the multivariate Cox proportional hazards regression. Among the eight NKG2D ligands, MICA (HR = 1.347; 95% CI = 1.012–1.793; p = 0.041), ULBP3 (HR = 0.453; 95% CI = 0.231–0.889; p = 0.021) and ULBP5 (HR = 3.617; 95% CI = 1.819–7.194; p

BackgroundGut microbes play an important role in the adaptation of insects. Polyphagous insects usually undergo changes in gut microbiota after host shift. The Bactrocera cucurbitae have a wide range of hosts, but the dynamic of gut microorganisms during host shift have not been thoroughly investigated. To understand the role of gut microbes in insect adaptation, it is necessary to study the dynamics of insect gut microorganisms during host transfer.MethodsUsing Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae) and its four hosts as study subjects, we investigated the dynamics of gut microbes during host transfer and the effects of different hosts on the gut microbial composition of B. cucurbitae.ResultsThe results showed that the Chao1 index of B. cucurbitae decreased significantly during host transfer, and the intestinal microorganisms were significantly affected by the original host, host, and generations. Furthermore, predicated changes in the abundance of secondary metabolite pathways after host transfer suggested that microorganisms may play an important role in the degradation of secondary metabolites, among which Providencia and Morganella have important functions in the gut of B. cucurbitae.ConclusionThis implied that microorganisms play a function in the host transfer process of B. cucurbitae and may be an important cofactor in the adaptation of B. cucurbitae to different hosts and environments, providing new research ideas for the future control of B. cucurbitae.

In recent decades, the increasingly widespread application of chemical pesticides has exacerbated the emergence of insecticide resistance among insect pests. In this study, we examined the rapid response of bacteria in the midgut of the fruit fly Bactrocera tau (Walker) (Diptera: Tephritidae) to stress induced by the insecticides lambda-cyhalothrin and spinosad by analyzing the bacterial community structure and diversity in the midguts of 4-day-old B. tau. The results revealed that 4-day-old B. tau females were more resistant to lambda-cyhalothrin and spinosad than their 4-day-old male counterparts. Alpha- and beta-diversity analyses revealed no significant differences between male and female B. tau with respect to the diversity and richness of gut bacteria in response to the same treatments. In response to treatment with lambda-cyhalothrin and spinosad at lethal concentration 50 (LC50), we detected significant changes in the structure and diversity of the bacterial community in the midguts of both male and female B. tau. Particularly among the dominant bacterial genera, there were decreases in the relative abundances of Citrobacter, Enterobacter, Klebsiella, and Pectobacterium. Increases were observed in the relative abundances of Dysgonomonas, Erwinia, and Providencia. Our findings provide a theoretical basis for gaining a better understanding of the relationships between midgut bacteria and the insecticide resistance of B. tau.

As an advantageous technique and service, the blockchain has shown great development and application prospects. However, its security has also met great challenges, and many security vulnerabilities and attack issues in blockchain-based services have emerged. Recently, security issues of blockchain have attracted extensive attention. However, there is still a lack of blockchain security research from a full-stack architecture perspective, as well as representative quantitative experimental reproduction and analysis. We aim to provide a security architecture to solve security risks in blockchain services from a full-stack architecture perspective. Meanwhile, we propose a formal definition of the full-stack security architecture for blockchain-based services, and we also propose a formal expression of security issues and defense solutions from a full-stack security perspective. We use ConCert to conduct a smart contract formal verification experiment by property-based testing. The security vulnerabilities of blockchain services in the Common Vulnerabilities and Exposures (CVE) and China Nation Vulnerability Database (CNVD) are selected and enumerated. Additionally, three real contract-layer real attack events are reproduced by an experimental approach. Using Alibaba's blockchain services and Identity Mixer in Hyperledger Fabric as a case study, the security problems and defense techniques are analyzed and researched. At last, the future research directions are proposed.

Tumor-infiltrating immune cells greatly participate in regulating tumorigenesis and metastasis of hepatocellular carcinoma (HCC). Natural killer cell, as an important role of innate immunity, plays an indispensable role in antitumor immunity and regulate tumor development. In this study, we firstly identified 251 NK cell marker genes of HCC based on single-cell RNA sequencing data. Subsequently, an NK cell marker genes-related prognostic signature (NKPS) was developed in the cancer genome atlas (TCGA) cohort for risk stratification and prognosis prediction. The predictive value of the NKPS in prognosis was well validated in different clinical subgroups and three external datasets (ICGC-LIHC cohort, GSE14520 cohort and Guilin cohort). Moreover, multivariate analysis revealed the independent prognostic value of NKPS for OS in HCC. Further functional analysis indicated the NKPS was associated with basic cellular processes, that may contribute to the development and progression of HCC. Thereafter, immune characteristics as well as the therapeutic benefits in NKPS risk score-defined subgroups were analyzed. Patients with low-risk score exhibited immune-active status, manifested as higher immune scores, more infiltration of CD8+ T cells and macrophage M1, and higher T-cell receptor (TCR) richness and diversity. Remarkably, the NKPS was negatively correlated with immunotherapy response-related signatures. In addition, the low-risk group exhibited significantly improved therapeutic benefits, either from immunotherapy or traditional chemotherapy and target therapy. Overall, the NKPS showed an excellent predictive value for prognosis and therapeutic responses for HCC, which might also provide novel insights into better HCC management strategies.

Low temperatures greatly influence newly introduced species, and increased cold tolerance can facilitate their establishment in new environments. The invasive alien species Ambrosia artemisiifolia is distributed at high latitudes and altitudes, where it suffers more from cold stress than it would at low latitudes or altitudes. Whether cold stress influences the accumulation of cryoprotectants and cold tolerance in A. artemisiifolia, and further influences the cold tolerance of its biological control agent, Ophraella communa, through feeding remain unknown. We investigated the levels of cryoprotectants and metabolic changes in A. artemisiifolia. We found that the level of total sugar, trehalose, proline, and other cold responsible metabolites increased in A. artemisiifolia after rapid cold-hardening (RCH) treatment, when compared to normal plants. These indicated that RCH treatment could improve the cold-hardiness of A. artemisiifolia. We then investigated the levels of cryoprotectants and metabolic changes in O. communa. We found that O. communa fed on RCH-treated A. artemisiifolia had higher levels of total sugar, trehalose, proline, glycerol, lipid, lower water content, lower super-cooling point, and increased cold tolerance compared to O. communa fed on normal A. artemisiifolia. This suggested that O. communa fed on cold-hardened A. artemisiifolia could increase its cold tolerance. Results showed a trophic transmission in insect cold tolerance. Our study enriches the theoretical basis for the co-evolution of cold tolerance in invasive and herbivorous insects.

Background Previous studies confirmed that most neoantigens predicted by algorithms do not work in clinical practice, and experimental validations remain indispensable for confirming immunogenic neoantigens. In this study, we identified the potential neoantigens with tetramer staining, and established the Co-HA system, a single-plasmid system coexpressing patient human leukocyte antigen (HLA) and antigen, to detect the immunogenicity of neoantigens and verify new dominant hepatocellular carcinoma (HCC) neoantigens.Methods First, we enrolled 14 patients with HCC for next-generation sequencing for variation calling and predicting potential neoantigens. Then, the Co-HA system was established. To test the feasibility of the system, we constructed target cells coexpressing HLA-A*11:01 and the reported KRAS G12D neoantigen as well as specific T-cell receptor (TCR)-T cells. The specific cytotoxicity generated by this neoantigen was shown using the Co-HA system. Moreover, potential HCC-dominant neoantigens were screened out by tetramer staining and validated by the Co-HA system using methods including flow cytometry, enzyme-linked immunospot assay and ELISA. Finally, antitumor test in mouse mode and TCR sequencing were performed to further evaluate the dominant neoantigen.Results First, 2875 somatic mutations in 14 patients with HCC were identified. The main base substitutions were C>T/G>A transitions, and the main mutational signatures were 4, 1 and 16. The high-frequency mutated genes included HMCN1, TTN and TP53. Then, 541 potential neoantigens were predicted. Importantly, 19 of the 23 potential neoantigens in tumor tissues also existed in portal vein tumor thrombi. Moreover, 37 predicted neoantigens restricted by HLA-A*11:01, HLA-A*24:02 or HLA-A*02:01 were performed by tetramer staining to screen out potential HCC-dominant neoantigens. HLA-A*24:02-restricted epitope 5'-FYAFSCYYDL-3' and HLA-A*02:01-restricted epitope 5'-WVWCMSPTI-3' demonstrated strong immunogenicity in HCC, as verified by the Co-HA system. Finally, the antitumor efficacy of 5'-FYAFSCYYDL-3'-specific T cells was verified in the B-NDG-B2mtm1Fcrntm1(mB2m) mouse and their specific TCRs were successfully identified.Conclusion We found the dominant neoantigens with high immunogenicity in HCC, which were verified with the Co-HA system.

Study region: Northwest of Guangxi Province, southwest China. Study focus: Knowing the role of isotopes in precipitation can improve understanding of climate change and eco-hydrological processes in complex critical-zone systems. However, knowledge of the influence of local meteorological factors on variations of precipitation isotopes remains limited as high-resolution rainwater sampling is lacking. New hydrological insights for the region: The temporal characteristics (intra-annual, seasonal, and monthly scales) of precipitation were analyzed using daily precipitation isotopes data from 2011 to 2020 and event-resolving data from 2019 to 2020 at Huanjiang in southwest China. The boosted regression tree model was used to explore the relationship between local meteorological factors and δ18O values. On an intra-annual scale, δ18O, δD, and d-excess values exhibited significant sinusoidal fluctuation, being lower and higher during the wet and dry seasons respectively. Temperature is the most sensitive meteorological factor to monsoon climate response. On the seasonal scale, ET0 explains 37.5% of isotopic variations in the dry season, thus indicating that the influence of evaporation on the rainwater isotopes variation should be considered. The “amount effect” of rainwater isotopes was clearly shown on the monthly scale. However, precipitation isotopes did not follow the “amount effect” on an inter-event scale, which can be explained by large-scale convection activity. These findings can improve interpretations of isotopic variations in precipitation in humid subtropical climate zones.

Zhenbao pill (ZBP) is a type of traditional Mongolian medicine that contains 241 ingredients such as flavonoids, anthraquinones, triterpenoids, sterols, and Calculus bovis, which are mainly used for treating neurological diseases in clinical practice. In this study, we established an Alzheimer’s disease (AD)-mimetic symptomatic zebrafish model by AlCl3 treatment and investigated the therapeutic effect of ZBP on the AD-mimetic zebrafish model. The results revealed that AlCl3 treatment led to behavioral impairment, induced apoptotic cell (body) in the brain, increased acetylcholinesterase (AChE) activity, and decreased the ACh content in zebrafish. After treatment with ZBP, 16 μg/mL ZBP could alleviate the behavioral disorder of zebrafish larvae. This effect was found superior to the effect of the cholinesterase inhibitor donepezil hydrochloride. The analysis of transcriptomics data revealed that ZBP mainly acted on apoptosis, the p53 signaling pathway, and the enrichment of the AChE-associated pathway. Furthermore, the overexpression of AChE RNA produced similar AD-mimetic disorders, whereas AChE-gene knockdown (AChE-MO) decreased the AChE activity and alleviated the AD-mimetic symptoms. These results suggested that ZBP is a potential cholinergic-enhancing drug, which can be used for the treatment of AD in the future.

BackgroundThe disease burden caused by chronic hepatitis B virus (HBV) infection is still heavy, and the current treatment scheme has not achieved a complete cure. Changes in natural and adaptive immunity usually accompany chronic HBV infection. As a marker expressed on dendritic cells (DCs), whether lysosome-associated membrane glycoprotein 3 (LAMP3) participates in chronic HBV infection deserves further analysis.MethodsWe retrieved chronic HBV infection transcriptional information from the Gene Expression Omnibus (GEO) database. The LAMP3 expression in the liver of patients with chronic hepatitis B (CHB) was analyzed in three GEO datasets and confirmed in our validation cohort (27 patients with CHB). Differentially expressed genes were obtained from one CHB cohort by comparing LAMP3high and LAMP3low expression subgroups. These genes underwent Gene Ontology, Kyoto Encyclopedia of Genes and Genomes analysis, and Gene Set Enrichment Analysis to decipher the influence of LAMP3 on the biological process and immunity changes in HBV infection. Furthermore, we investigated the potential relationship between LAMP3 levels, the abundance of infiltrating immune cells, and liver dysfunction.ResultsCompared to healthy controls, LAMP3 expression was upregulated in the transcriptional profiles of the liver in patients with CHB. The high LAMP3 expression was related to T cell activation and the chemokine signaling pathway. The LAMP3 gene was positively linked to marker sets of infiltrating activated regulatory T cells (Treg), T cell exhaustion, monocytes, and DCs. Moreover, CHB patients with high LAMP3 expression had unfavorable liver dysfunction.ConclusionsLAMP3 is a gene related to HBV infection, which might be involved in HBV infection by regulating T cell activation and adaptive immune response.

Heat shock proteins (Hsps) function as molecular chaperones that enable organisms to withstand stress and maintain normal life activities. In this study, we identified heat shock protein 70 (encoded by hsp70), which exhibits a higher expression in the mature male testis than in the unmature testis of Ophraella communa. Tissue expression profile revealed that Ochsp70 levels in males were highest in the testis, whereas those in females were highest in the head. Moreover, the expression of Ochsp70 was found to be significantly induced in female bursa copulatrix after mating. Double-stranded RNA dsOchsp70 was injected into males to performance RNA interference, which significantly decreased the male Ochsp70 expression levels within 20 d post-injection, whereas no effect was observed on the Ochsp70 expression level in the females after mating with dsOchsp70-injected males. However, significant downregulation of female fertility was marked simultaneously. Furthermore, knockdown of female Ochsp70 expression also led to a significant reduction in fertility. Finally, comparative transcriptomic analysis identified glucose dehydrogenase and insulin-like growth factor binding protein as putative downstream targets of Ochsp70. Overall, we deduced that Ochsp70 is an indispensable gene and a potential male mating factor in O. communa, which regulates reproduction.

Abstract Concentrations of heavy metals continue to increase in soil environments as a result of both anthropogenic activities and natural processes. Cadmium (Cd) is one of the most toxic heavy metals and poses health risks to both humans and the ecosystem. Herein, we explore the impacts of Cd on a soil–plant system composed of oilseed rapes (Brassica napus and Brassica juncea) and bacteria. The results showed that Cd accumulation within tissues of two species of oilseed rapes enhanced with increasing concentrations of Cd in soils, and Cd treatment decreased their chlorophyll content and suppressed rapeseeds growth. Meanwhile, Cd stress induced the changes of antioxidative enzymes activities of both B. napus and B. juncea. Response to Cd of bacterial community was similar in soil-two species of oilseed rapes system. The impact of Cd on the bacterial communities of soils was greater than bacterial communities of plants (phyllosphere and endophyte). The α-diversity of bacterial community in soils declined significantly under higher Cd concentration (30 mg/kg). In addition, soil bacterial communities composition and structure were altered in the presence of higher Cd concentration. Meanwhile, the bacterial communities of bulk soils were significantly correlated with Cd, while the variation of rhizosphere soils bacterial communities were markedly correlated with Cd and other environmental factors of both soils and plants. These results suggested that Cd could affect both the growth of plants and the indigenous bacterial community in soil–plant system, which might further change ecosystem functions in soils.

Abstract Background Since it’s a challenging task to precisely predict the prognosis of patients with hepatocellular carcinoma (HCC). We developed a nomogram based on a novel indicator GMWG [(Geometric Mean of gamma-glutamyltranspeptidase (GGT) and white blood cell (WBC)] and explored its potential in the prognosis for HCC patients. Methods The patients enrolled in this study were randomly assigned to training and validation cohorts. And we performed the Least Absolute Shrinkage and Selection Operator proportional hazards model (LASSO Cox) model with clinical characteristics, serum indexes, and novel GMWG. Multivariate analysis was performed to build a nomogram. The performance of the nomogram was evaluated by C-index, the area under the receiver operating characteristic curve (AUC), and the calibration curve. Kaplan-Meier curves showed discrimination of the nomogram. Clinical utility was assessed by decision curve analysis (DCA). The discrimination ability of the nomogram was determined by the net reclassification index (NRI). Results The geometric mean of GGT and white WBC count (GMWG), neutrophil to lymphocyte ratio (NLR), and tumor size were significantly associated with the overall survival (OS). The variables above were used to develop the nomogram. The indexes of nomogram were 0.70 and 071 in the training or validation cohort, respectively. AUC of 1-, 3- and 5-year OS showed satisfactory accuracy as well. The calibration curve showed agreement between the ideal and predicted values. Kaplan-Meier curves based on the overall survival (OS) and disease-free survival (DFS) showed significant differences between nomogram predictive low and high groups. DCA showed clinical utilities while NRI showed discrimination ability in both training or validation cohort. Conclusions GMWG might be a potential prognostic indicator for patients with HCC. The nomogram containing GMWG also showed satisfaction prediction capacity.

To predict and mitigate the effects of climate change on communities and ecosystems, the joint effects of extreme climatic events on species interactions need to be understood. Using the common ragweed (Ambrosia artemisiifolia L.)—leaf beetle (Ophraella communa) system, we investigated the effects of heat wave and elevated CO2 on common ragweed growth, secondary metabolism, and the consequent impacts on the beetle. The results showed that elevated CO2 and heat wave facilitated A. artemisiifolia growth; further, A. artemisiifolia accumulated large amounts of defensive secondary metabolites. Being fed on A. artemisiifolia grown under elevated CO2 and heat wave conditions resulted in the poor performance of O. communa (high mortality, long development period, and low reproduction). Overall, under elevated CO2, heat wave improved the defensive ability of A. artemisiifolia against herbivores. On the other hand, enhanced adaptability to climatic changes may aggravate invasive plant distribution, posing a challenge to the control of invasive plants in the future.

BackgroundAlthough effective vaccines have been developed against coronavirus disease 2019 (COVID-19), the level of neutralizing antibodies (NAbs) induced after vaccination in the real world is still unknown. The aim of this work was to evaluate the level and persistence of NAbs induced by two inactivated COVID-19 vaccines in China.MethodsSerum samples were collected from 1,335 people aged 18 years and over who were vaccinated with an inactivated COVID-19 vaccine at Peking University People’s Hospital from January 19 to June 23, 2021, for the detection of anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies.ResultsThe positive rate for NAbs against SARS-CoV-2 was 79–91% from the first month to the second month after the second vaccine dose. The gradual decline in positivity rate for NAb response was observed from 78% at 3 months post-vaccination to 0% at 12 months post-vaccination. When there was a 21-day interval between the two doses of vaccine, the NAb positivity rate was 0% 6 months after the second dose. NAb levels were significantly higher when the interval between two doses were 3–8 weeks than when it was 0–3 weeks (χ2 = 14.04, p < 0.001). There was a linear correlation between NAbs and IgG antibodies in 1,335 vaccinated patients. NAb levels decreased in 31 patients (81.6%) and increased in 7 patients (18.4%) over time in the series of 38 patients after the second vaccination. The NAb positivity rate was significantly higher in 18- to 40-year-old subjects than in 41- to 60-year-old subjects (t = −1.959, p < 0.01; t = 0.839, p < 0.01).ConclusionThe NAb positivity rate was the highest at the first and second month after the second dose of vaccine, and gradually decreased over time. With a 21-day interval between two doses of vaccine, neutralizing antibody levels persisted for only 6 months after the second dose of vaccine. Therefore, a third vaccine dose is recommended. Our results suggest that in cases in which NAbs cannot be detected, IgM/IgG antibodies can be detected instead. The level of NAbs produced after vaccination was affected by age but not by sex. Our results suggest that an interval of 21 to 56 days between shots is suitable for vaccination.

Abstract Background Hepatocarcinogenesis is reportedly correlated with abnormal m6A modifications; however, it is unknown whether m6A RNA methylation regulators facilitate the occurrence of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). Thus, we constructed an m6A-related model that may enhance HBV-related HCC prognosis. Methods Gene signatures of HNRNPA2B1 and RBM15 were generated by univariate and Lasso Cox regression analyses using the gene set and clinical information from The Cancer Genome Atlas (TCGA) database. High-risk and low-risk groups were confirmed based on the gene signature model. Furthermore, we validated the predictive roles of the two genes for overall survival (OS) in the GSE14520 dataset. The relative expression of 22 paired mRNAs was measured using quantitative real-time polymerase chain reaction (qRT-PCR) analysis to determine whether the two genes had a predictive role in our Guilin cohort. Results The differences in OS between the high-risk and low-risk groups were statistically significant in the TCGA (p = 0.003) and GSE14520 (p = 0.045) datasets, but not in the Guilin cohort, owing to differences in clinical information among the three cohorts (mainly the TNM stage and survival state). Stratified analysis of TNM stages showed that the two-gene signature acted as a prognostic indicator of HBV-related HCC patients in the early TNM stage; both TCGA and GSE14520 cohorts showed statistical significance. Moreover, multivariate Cox regression analysis indicated that the two-gene signature was an independent factor for predicting prognosis (HR = 1.087, 95% CI: 1.007–1.172). Correlation analysis between the gene signature and clinical features revealed that the risk stratification was significantly correlated with grade and survival state. Finally, Gene Set Enrichment Analysis (GSEA) revealed that the KEGG pathways associated with the cell cycle, DNA replication, the spliceosome, repair, and metabolism-related processes were all significantly enriched in the high-risk group. Among the enriched genes, the expression levels of the replication protein RPA1 and the pre-mRNA splicing factor SF3B1 were significantly upregulated in the high-risk group. These results might help in elucidating the underlying molecular mechanisms of HBV-related HCC. Conclusions Our data may provide new predictive signatures and potential therapeutic targets to identify and treat HBV-related HCC patients in the early disease stage.

Chemokines are a family of cytokines that orchestrate the migration and positioning of immune cells within tissues and are critical for the function of the immune system. CCR2 participates in liver pathology, including acute liver injury, chronic hepatitis, fibrosis/cirrhosis, and tumor progression, by mediating the recruitment of immune cells to inflammation and tumor sites. Although a variety of chemokines have been well studied in various diseases, there is no comprehensive review presenting the roles of all known chemokine ligands of CCR2 (CCL2, CCL7, CCL8, CCL12, CCL13, CCL16, and PSMP) in liver disease, and this review aims to fill this gap. The introduction of each chemokine includes its discovery, its corresponding chemotactic receptors, physiological functions and roles in inflammation and tumors, and its impact on different immune cell subgroups.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) shows a high degree of homology with SARS-CoV. They share genes, protein sequences, clinical manifestations, and cellular entry patterns. Thus, SARS research may serve helpful in gaining a better understanding of the current coronavirus disease 2019 (COVID-19) pandemic. Serum antibodies from convalescent patients with SARS collected in 2018 were used to target the recombinant SARS-CoV-2 spike protein via a chemiluminescence microsphere immunoassay. Antibodies of convalescent patients with SARS exhibited serous immune cross-reactivity with the SARS-CoV-2 spike protein. The serous antibodies, excluding S22 of convalescent patients with SARS, did not competitively inhibit the binding of SARS-CoV-2 spike protein to ACE2. T cellular immunity research was conducted in vitro using peripheral blood mononuclear cells (PBMCs) stimulated by pooled peptide epitopes 15 years post-infection. Interferon gamma was detected and the PBMC transcriptomic profile was obtained. The heatmap of the transcriptomic profile showed that mRNAs and circRNAs of the SARS group clustered together after being stimulated by the peptide epitope pool. Differentially expressed mRNAs were most significantly enriched in immunity and signal transduction (P < 0.01). SARS elicits cytokine and chemokine responses, partially consistent with previously published data about COVID-19. Overall, our results indicate that antibodies from convalescent patients with SARS persisted for 15 years and displayed immune cross-reactivity with the SARS-CoV-2 spike protein. The immune status of patients with SARS 15 years post-infection may provide a better understanding of the future immune status of patients with COVID-19.

In February 2020, an inpatient in Peking University People's Hospital (PKUPH), China, was confirmed positive for the novel coronavirus. In this case, 143 hemodialysis patients were labeled as close contacts and required to be placed under the hospital-based group medical quarantine (HB-GMQ) for 2 weeks by the authorities. After the case was reported, false or misleading information about the case flourished on social media platforms, which led to infodemic. Under this context, PKUPH adopted patient-centered humanistic care to implement the HB-GMQ, through the synergy of administrative, healthcare, logistical, and other measures under the model of patient-centered care of the Massachusetts Medical Society (MMS). As a result, all the patients tided over the HB-GMQ with no COVID-19 infection and no unanticipated adverse events, and all met the criteria for lifting the HB-GMQ. According to the questionnaires taken during the HB-GMQ, a high level of satisfaction was found among the quarantined and no symptomatic increase of anxiety and depression in the patients before and during the HB-GMQ, by comparing the Zung self-rating anxiety scale (SAS) and self-rating depression scale (SDS) conducted in December 2019 and on the 12th day of the HB-GMQ. This article is to brief on PKUPH's experience in implementing patient-centered humanistic care tailored to hemodialysis patients under the HB-GMQ, and to validate the hypothesis that patient-centered humanistic care is effective and helpful to help them tide over the HB-GMQ, so as to shed light on how to implement the HB-GMQ and cope with the HB-GMQ-induced problems in other hospitals.

PD-(L)1 inhibitor could improve the survival of locally advanced esophageal cancer (ESCA) patients, but we cannot tailor the treatment to common biomarkers. WNT signaling activation was associated with primary resistance to immunotherapy. In this study, we used our two clinical cohorts (BJCH n = 95, BJIM n = 21) and three public cohorts to evaluate and verify a new immunotherapeutic biomarker based on WNT signaling in ESCA patients. Our findings showed that WNT signaling-related genes stratified TCGA patients into Cluster 1, 2, and 3, among which, Cluster 3 had the worst prognosis. The most up- and down-regulated genes in Cluster 3 were IGFBP1 and WNT3A. Further analysis validated that IGFBP1hiWNT3Alo ESCA patients had significantly poor RFS and OS in the TCGA and BJCH cohorts. Interestingly, IGFBP1hiWNT3Alo patients had a good response and prognosis with immunotherapy in three independent cohorts, exhibiting better predictive value than PD-L1 expression (signature AUC = 0.750; PD-L1 AUC = 0.571). Moreover, IGFBP1hiWNT3Alo patients may benefit more from immunotherapy than standard treatment (p = 0.026). Immune cell infiltration analysis revealed a significant increase in DC infiltration in IGFBP1hiWNT3Alo patients post-immunotherapy (p = 0.022), which may enhance immune response. The IGFBP1hiWNT3Alo signature could predict patients who benefited from PD-(L)1 inhibitor treatment and may serve as a biomarker in ESCA.

The beetle Ophraella communa is an effective biological control agent against the invasive common ragweed spread across various ecosystems with variable temperature ranges. The trend in climate change attributed to fluctuating temperatures and abrupt rainfalls is expected to continue. This study aimed to better understand the effects of thermal fluctuation on O. communa by exposing all their life stages to heat stress under different treatments. Repeated exposure to high temperatures, relative to constant milder temperatures, increased the duration of immature development, mean generation time, and the adult longevity, decreased the intrinsic rate of increase, finite rate of population increase, net reproductive rate, survival rate, overall longevity, body length, and mass of adults and positively affected overall fecundity by prolonging the oviposition period, biasing sex ratio towards females. After exposure to heat stress, the mating success and production of viable offspring were higher in O. communa. Our findings demonstrate that exposure to heat stress negatively affects ragweed beetles, but they were able to survive and reproduce.

At the end of 2019, an outbreak of pneumonia took place caused by a new coronavirus (SARS-CoV-2 virus), named coronavirus disease 2019 (COVID-19). A series of strict prevention and control measures were then implemented to reduce the spread of the epidemic. Influenza, another respiratory tract virus, may also respond to these measures. To assess the impact of these measures, we used the total number of passengers movement in mainland China from 2018 to 2020 and daily number of railway passenger flow during the 2020 Spring Festival travel rush to reflect the population movement and to analyze newly and cumulatively confirmed COVID-19 and influenza cases. We found that implementing the series of measures against COVID-19 mitigated both COVID-19 and influenza epidemics in China. Prevention and control measures for COVID-19 might be used to control respiratory tract infections to reduce the national health economic burden caused by these pathogens.

Abstract Antennal olfaction plays a key role in insect survival, which mediates important behaviors like host search, mate choice, and oviposition site selection. As an oligophagous insect, olfaction is extremely important for Ophraella communa to locate host plants. However, information on the olfactory genes has been lacking in O. communa. Using next generation sequencing, we assembled the antennal transcriptome of O. communa and first reported the major chemosensory genes necessary for olfaction in this species. In this study, a total 105 candidate chemosensory genes were identified in O. communa antennae, including 25 odorant-binding proteins (OBPs), 11 chemosensory proteins (CSPs), four sensory neuron membrane proteins (SNMPs), 30 odorant receptors (ORs), 18 ionotropic receptors (IRs), and 17 gustatory receptors (GRs). We also identified full-length sequences of the highly conserved ORco and IR8a/25a family in O. communa. In addition, the expression profile of 15 ORs and four OBPs were validated by quantitative real-time polymerase chain reaction (qPCR). We found that OcomOR2/4/19 and OcomOBP19/20 had a biased expression in male antennae, and OcomOR8 had a biased expression in the female antennae. This large number of chemosensory genes handled by homology analysis and qPCR results will provide the first insights into molecular basis for the olfactory systems of O. communa as well as advance our understanding of olfactory mechanisms in Coleoptera.

Low-rate Denial of Service (LDoS) attack is a special DoS attack. The routing protocol is vulnerable to many types of attacks in a wireless sensor network (WSN), which is an important network type of the Internet of Things (IoT). The novel LDoS attack to the routing protocol is proposed to evaluate the security and trust mechanism in the WSN. In fact, the LDoS attack is difficult to be detected due to its small-signal characteristics, so it is a serious threat to the security and trust of the WSN. A Hilbert-Huang transform (HHT) time-frequency joint analysis approach is utilized to analyze the non-stationary small signal that is produced by the LDoS attack. However, false intrinsic mode function (IMF) components are the challenge problems to precisely detect the LDoS attack. Correlation coefficient and Kolmogorov-Smirnov (KS) test approaches are united to evaluate the trustworthy of IMF components and exclude the false IMF components. Hilbert-Huang transformation and trust evaluation approaches are combined to detect the novel LDoS attack in Zigbee WSN. CC2530 system-on-chip integrated with ZigBee protocol is utilized to build a wireless sensor node. Random routing REQuest (RREQ) flooding attack is used to implement the routing layer LDoS attack in Zigbee WSN. If the correlation coefficient value of IMF component relative to original traffic is more than 0.3 and the KS similarity probability value of the IMF component relative to the original traffic is more than 0.4, the IMF component is identified as high trust IMF components that will be used to detect LDoS attack. If the IMF component only satisfies one of the trust evaluation conditions, the IMF component is identified as low trust IMF component. Otherwise, the IMF component is the false IMF component. We have proposed a scalable LDoS attack detection architecture for both WSN and IoT. The experimental results demonstrate that the novel approach is highly effective to detect the LDoS attack in the ZigBee WSN.

Background: Repeat hepatectomy is an important treatment for patients with repeat recurrent hepatocellular carcinoma (HCC).Methods: This study was a multicenter retrospective analysis of 1,135 patients who underwent primary curative liver resection for HCC. One hundred recurrent patients with second hepatectomy were included to develop a nomogram to predict the risk of post-recurrence survival (PRS). Thirty-eight patients in another institution were used to externally validate the nomogram. Univariate and multivariate Cox regression analyses were used to identify independent risk factors of PRS. Discrimination, calibration, and the Kaplan–Meier curves were used to evaluate the model performance.Results: The nomogram was based on variables associated with PRS after HCC recurrence, including the tumor, node, and metastasis (TNM) stage; albumin and aspartate aminotransferase levels at recurrence; tumor size, site, differentiation of recurrences; and time to recurrence (TTR). The discriminative ability of the nomogram, as indicated by the C statistics (0.758 and 0.811 for training cohort and external validation cohorts, respectively), was shown, which was better than that of the TNM staging system (0.609 and 0.609, respectively). The calibration curves showed ideal agreement between the prediction and the real observations. The area under the curves (AUCs) of the training cohort and external validation cohorts were 0.843 and 0.890, respectively. The Kaplan–Meier curve of the established nomogram also performed better than those of both the TNM and the BCLC staging systems.Conclusions: We constructed a nomogram to predict PRS in patients with repeat hepatectomy (RH) after repeat recurrence of HCC.

Microbial communities associated with the plant phyllosphere and endosphere can have both beneficial as well as detrimental effects on their hosts. There is an ongoing debate to which extend the phyllosphere and endosphere microbiome assembly is controlled by the host plant how pronounced cultivar effects are. We investigated the bacterial and fungal communities from the phyllosphere and endosphere of 10 different rice cultivars grown under identical environmental conditions in the frame of a targeted approach to identify drivers of community assembly. The results indicated that the endophytic bacterial communities were clearly separated into two groups. The α-diversity and microbial network complexity within Group I were significantly lower than in Group II. Moreover, the genera Nocardioides, Microvirga, and Gaiella were significantly more abundant in Group II and only present in the interaction networks of this group. These three genera were significantly correlated with α- and β-diversity of the endophytic bacterial community and thus identified as major drivers of the endosphere community. We have identified keystone taxa that shape endophytic bacterial communities of different rice cultivars. Our overall findings provide new insights into plant-microbe interactions, and may contribute to targeted improvements of rice varieties in the future.

BackgroundHepatocellular carcinoma (HCC) is the seventh most common malignancy and the second most common cause of cancer-related deaths. Autophagy plays a crucial role in the development and progression of HCC.MethodsUnivariate and Lasso Cox regression analyses were performed to determine a gene model that was optimal for overall survival (OS) prediction. Patients in the GSE14520 and GSE54236 datasets of the Cancer Genome Atlas (TCGA) were divided into the high-risk and low-risk groups according to established ATG models. Univariate and multivariate Cox regression analyses were used to identify risk factors for OS for the purpose of constructing nomograms. Calibration and receiver operating characteristic (ROC) curves were used to evaluate model performance. Real-time PCR was used to validate the effects of the presence or absence of an autophagy inhibitor on gene expression in HepG2 and Huh7 cell lines.ResultsOS in the high-risk group was significantly shorter than that in the low-risk group. Gene set enrichment analysis (GSEA) indicated that the association between the low-risk group and autophagy- as well as immune-related pathways was significant. ULK2, PPP3CC, and NAFTC1 may play vital roles in preventing HCC progression. Furthermore, tumor environment analysis via ESTIMATION indicated that the low-risk group was associated with high immune and stromal scores. Based on EPIC prediction, CD8+ T and B cell fractions in the TCGA and GSE54236 datasets were significantly higher in the low-risk group than those in the high-risk group. Finally, based on the results of univariate and multivariate analyses three variables were selected for nomogram development. The calibration plots showed good agreement between nomogram prediction and actual observations. Inhibition of autophagy resulted in the overexpression of genes constituting the gene model in HepG2 and Huh7 cells.ConclusionsThe current study determined the role played by autophagy-related genes (ATGs) in the progression of HCC and constructed a novel nomogram that predicts OS in HCC patients, through a combined analysis of TCGA and gene expression omnibus (GEO) databases.

Soil erosion has been a major environmental issue in young citrus orchards in karst areas of Southwest China. Straw mulching is an effective measure to reduce soil erosion. However, few studies have considered this measure in soil and water conservation in citrus orchards in karst areas. In this study, the effects of straw mulching on runoff and sediment reduction in young citrus orchards were studied using the simulated rainfall method with two typical soils—red soil and lime soil—in karst areas. This study consisted of two rainfall intensities (60 and 120 mm/h) and four straw coverages (0, 20, 50, and 80%). The results showed that straw mulching can significantly reduce surface soil loss in both lime soil and red soil. The surface runoff reduction rate of lime soil with straw mulching was much higher than that of red soil. Under the condition of heavy rainfall (120 mm/h), the surface soil loss rate was reduced by more than 57.5% in lime soil with 20% straw mulching. In red soil, a similar reduction rate of soil loss can only be achieved when straw mulching is 50%. Straw mulching reduced the surface soil loss of lime soil in young citrus orchards, mainly because of limited runoff transport, while red soil was limited by soil stripping. The results can provide a scientific basis for soil and water conservation in young citrus orchards in karst areas.

Coarse root systems provide a framework for water and nutrient absorption from the soil and play an important role in plant survival in harsh environments. However, the adaptions of plant roots in soil-limited environments with low water storage capacity and nutrient content needs to be better understood. The adaptation strategies of two common plant species (a deciduous tree Platycarya longipes and an evergreen shrub Tirpitzia ovoidea) from two contrasting habitats (a shallow rocky soil and a nearby deep soil) in a karst region of subtropical China were compared and analyzed. Foliar nutrient concentrations, stoichiometry, stable carbon, and oxygen isotopes were used to determine plant nutrient and water use status across these two habitats. Six indexes, including maximum root depth, maximum root radial extent, number of major roots and secondary roots, and tapering rate and curvature, were all investigated to characterize coarse root systems. Results show that both species exhibited similar nutrient and water use status in the two habitats that had contrasting water holding capacity and available nutrient content. On the other hand, although maximum root depths of the individual plants were not deeper than 33 cm, maximum radial extents were much larger when compared to rooting depths. Specifically, the ratio of radial extent to depth in the soil-limited habitat was approximately 6 and 1.5 times higher than that in the deep-soil habitat for the tree and shrub, respectively. Additionally, especially for the tree, a larger root radial extent was further accompanied by lower root tapering rate and bending levels. Our results provided evidence that plants growing in soil-limited environments maintain a stable resource use status along with large radially extended coarse root systems in humid karst regions like southwest China.

Ophraella communa is an effective bio-control agent of the invasive common weed. By now, the reference genes in O. communa have not yet been screened and validated. The aim of this study was to screen for the most stable reference genes in different backgrounds, such as different developmental stages, sexes, tissues, and male reproductive system with different body sizes. We selected 12 common housekeeping genes involved in different biological processes, including GAPDH, ACT1, ACT2, ARF1, ARF4, SDH, βTUBC, RPL4, RPL19, RPS18, EF1α, and COX as the candidate reference genes. To analyze the stability of the candidate reference genes, we first used three dedicated algorithms, GeNorm, NormFinder, and BestKeeper, and further comprehensive ranking was provided by ReFinder. The results showed that RPL19 and RPL4 exhibited the least variation in different developmental stages/sexes and in male reproductive systems with different body sizes. COX proved to be most suitable for normalizing the gene expression levels in different tissues, and coincidentally, RPL19 was also found to be second in terms of stability in this study. To the best of our knowledge, this is the first study to identify suitable reference genes for analyzing gene expression in O. communa; thus, this study would lay the foundation for future research on the molecular physiology and biochemistry of O. communa and other insects.

The inevitable integration of critical infrastructure to public networks has exposed the underlying industrial control systems to various attack vectors. In this paper, we model multi-stage crypto ransomware attacks, which are today an emerging cyber threat to critical infrastructure. We evaluate our modeling approach using multi-stage attacks by the infamous WannaCry ransomware. The static malware analysis results uncover the techniques employed by the ransomware to discover vulnerable nodes in different SCADA and production subnets, and for the subsequent network propagation. Based on the uncovered artifacts, we recommend a cascaded network segmentation approach, which prioritizes the security of production network devices. Keywords: Critical infrastructure, Cyber-attack, Industrial control system, Crypto ransomware, Vulnerability

Abstract Background Identification of novel MDM2 or p53 binding proteins may reveal undefined oncogenes, tumor suppressors, signaling pathways and possible treatment targets. Methods By means of immunoprecipitation and Mass Spectrometry analysis, we aimed to identify novel regulators of the MDM2-p53 pathway. We further clarified the impact of MYL6B on the p53 protein level and on the process of apoptosis. We also investigated the role of MYL6B in hepatocellular carcinoma by clone formation assay and by determining the correlation between its expression and prognosis of HCC patients. Results We identified a novel MDM2 and p53 binding protein, MYL6B. It is a myosin light chain that could bind myosin II heavy chains to form non-muscle myosin II holoenzymes (NMII). We found that MYL6B could facilitate the binding of MDM2 to p53, which consequently promotes the ubiquitination and degradation of p53 protein. We further proved that MYL6B exerts the suppression effect on p53 as part of NMII holoenzymes because inhibiting the ATPase activity of myosin II heavy chain largely blocked this effect. We also discovered that MYL6B is overexpressed in HCC tissues and linked to the bad prognosis of HCC patients. Knocking out of MYL6B dramatically suppressed the clonogenic ability and increased the apoptosis level of HCC cell lines. Conclusions To summary, our results demonstrate that MYL6B is a putative tumor driver gene in HCC which could promote the degradation of p53 by enhancing its’ MDM2-mediated ubiquitination.

Abstract Soil water movement is difficult to explain with event-scale approaches, especially in karst regions. This paper focuses on investigating seasonal recharge and mean residence time (MRT) of soil water based on temporal variation of stable isotopes (δD and δ18O) and a dispersion model (DM), and discussing their differences along a footslope and a depression in a small karst catchment of southwest China. Temporal variations of the stable isotopes in precipitation and soil water within 0–100 cm profiles were monitored weekly for approximately 43 and 99 weeks. Results show that the seasonal recharge of soil water inside the footslope and the depression were similar, but the vertical flow velocity was higher implying a faster hydrological process in the footslope. The MRT of soil water (2–64 weeks) increased roughly, suggesting decreasing velocity of water displacement with increasing depth. However, the MRT at 60–100 cm depths in the depression (47–64 weeks) was obviously longer than at other sites, revealing more intensive water mixing. Furthermore, a shallower isotopic damping depth was found in the depression, indicating stronger delay and attenuation effects on base flow recharge. These results provide new insights into research on hydrological processes in karst areas.

Chemosensory proteins (CSPs) are considered to be the transporter linking odorant chemicals and receptors on sensory neurons. However, the extensive expression patterns of CSPs in insects suggest that CSPs are also involved in other physiological processes; the range of their functions, however, remains uncertain. In this study, we successfully characterized and cloned the CSP12 of Ophraella communa (OcomCSP12). The open reading frame of OcomCSP12 encodes 131 amino acids, with four conserved cysteine residues. The expression patterns of OcomCSP12 validated by quantitative real-time PCR (qRT-PCR) showed that OcomCSP12 is specifically expressed in female ovary. Furthermore, compared with the control treatment, silencing OcomCSP12 resulted in significantly reduced oviposition in females. Surprisingly, the knock-down rate of OcomCSP12 exceeded 95% and remained depressed for more than 15 days, indicating that RNA interference (RNAi) was a suitable method for exploring the function of CSP12 in O. communa. These findings increase our understanding of the expression profile and function of the CSP gene family in insects.

Considering the predicted rising temperatures under current climate change and heat wave scenarios, organisms are expected to suffer more intense and frequent thermal stress. Induced heat is accumulated by organisms and can cause a variety of physiological stress responses. Ophraella communa is an effective biological control agent of common ragweed, Ambrosia artemisiifolia, but the responses of this biocontrol agent to heat stress have not been fully elucidated and, therefore, its potential responses to climate change are uncertain. We investigated the physiological metabolism of subsequent O. communa adults after: (1) different developmental stages (egg, larval, pupal, and adult) were exposed to thermal stress for 3 h each day for 3, 5, 5, and 5 days, respectively (by stage); and (2) individuals were exposed to thermal stress throughout the egg-to-adult period for 3 h each day. The high temperatures of 40, 42, and 44°C were used to induce thermal stress. A control group was reared at 28 ± 2°C. The results showed that short- or long-term exposure to daily phasic high temperatures significantly decreased water and lipid contents and significantly increased glycogen and glycerol contents in all adults (i.e., after exposure of different stages or throughout the egg-to-adult period). However, the total sugar content significantly increased in adults after the eggs and larvae were exposed to brief short-term thermal stress. Compared to the control, the total sugar content was also significantly higher in the adults and pupae exposed to 44°C. Total sugar content in females increased significantly in response to long-term phasic thermal stress at 40°C. However, sugar content of males exposed to 44°C decreased significantly. After long-term phasic thermal stress, water and glycogen contents in males were significantly higher than in females; however, females had higher total sugar and lipid contents. Therefore, our study provides a basic understanding of the metabolic responses of O. communa to thermal stress and offers insights into its potential as a natural biocontrol agent against A. artemisiifolia during the summer season and under predicted climate change scenarios.

In karst regions, soil architecture varies along topographical locations, resulting in marked differences in infiltration rates. However, the relationship between soil architecture and preferential flow (PF) is still unclear. In this study, dye tracing was used to investigate PF and the dominant effects of five types of soil architecture in a small karst catchment. These soil architectures included deep clay soil in farmland (CSF) and deep clay soil in shrubland (CSS) in the depression; shallow clay soil with minimally weathered, slanted bedrock on downslope locations (CSWD); shallow sandy loam soil with highly weathered bedrock on midslope locations (SLSM); and sandy loam soil with rock fragments on upslope locations (SLSU). The results showed that macropores and cracks were the major channels that resulted in a high degree of PF in depressions and downslope locations. Preferential flow also continued along the rock–soil interface in the downslope locations. Finger flow mainly appeared in mid- and upslope locations, and cracks mainly appeared in the surface soil layer. However, down- and upslope locations showed lower PF. We observed that PF might not contribute to the different infiltration rates in different topographical locations. Tillage in depressions damages macropores, thereby minimizing vertical percolation. The high saturated hydraulic conductivity () of the soil matrix covered the development of PF in the mid- and upslope locations, and the decrease in weakened the formation of PF as a result of heterogeneity of soil properties. These findings demonstrate that infiltration-excess runoff may occur in depressions and downslope locations and that saturation-excess runoff may occur in mid- and upslope locations.

Ambrosia artemisiifolia is a noxious invasive alien weed, that is harmful to the environment and human health. Ophraella communa is a biocontrol agent for A. artemisiifolia, that was accidentally introduced to the Chinese mainland and has now spread throughout southern China. Recently, we found that upon artificial introduction, O. communa can survive in northern China as well. Therefore, it is necessary to study the cold hardiness of O. communa. Many genes have been identified to play a role in cold-tolerance regulation in insects, but the function of the gene encoding non-muscle myosin regulatory light chain (MRLC-sqh) remains unknown. To evaluate the role played by MRLC-sqh in the cold-tolerance response, we cloned and characterized MRLC-sqh from O. communa. Quantitative real-time PCR revealed that MRLC-sqh was expressed at high levels in the gut and pupae of O. communa. The expression of MRLC-sqh was shown to decrease after cold shock between 10 and 0 °C and ascend between 0 and −10 °C, but these did not show a positive association between MRLC-sqh expression and cold stress. Silencing of MRLC-sqh using dsMRLC-sqh increased the chill-coma recovery time of these beetles, suggesting that cold hardiness was reduced in its absence. These results suggest that the cold hardiness of O. communa may be partly regulated by MRLC-sqh. Our findings highlight the importance of motor proteins in mediating the cold response in insects.

Ophraellacommuna LeSage is an effective biological control agent of common ragweed, Ambrosia artemisiifolia L., which competes with crops and causes allergic rhinitis and asthma. However, thermal stress negatively affects the developmental fitness and body size of this beetle. High temperatures cause a variety of physiological stress responses in insects, which can cause oxidative damage. We investigated the total protein content and activity of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), and peroxidases (PODs) in O. communa adults when its different developmental stages were exposed to high temperatures (40, 42, and 44°C) for 3 h each day for 3, 5, 5, and 5 days, respectively (by stage), and a whole generation to high temperatures (40, 42, and 44°C) for 3 h each day. A control group was reared at 28 ± 2°C. Under short-term daily phasic high-temperature stress, total protein contents were close to the control as a whole; overall, SOD activities increased significantly, CAT activities were closer to or even higher than the control, POD activities increased at 40°C, decreased at 42 or 44°C; stage-specific response was also observed. Under long-term daily phasic high-temperature stress, total protein content increased significantly at 44°C, SOD activities increased at higher temperatures, decreased at 44°C; CAT activities of females increased at ≤42°C, and decreased at 44°C, CAT activities of males decreased significantly; POD activities of females increased at 40°C, decreased at ≥42°C, POD activities of males decreased at 44°C; and antioxidant enzymes activities in females were significantly higher than those in males. Antioxidative enzymes protect O. communa from oxidative damage caused by thermal stress.

Chill tolerance plays a crucial role that allows insect species to adapt to cold environments. Two Chinese geographical populations (Laibin and Yangzhou populations) were selected to understand the chill resistance and evolutionary potential in the Ophraella communa, a biological control agent of the invasive common ragweed, Ambrosia artemisiifolia. Super-cooling point assays, knockdown tests under static low-temperature conditions and determination of glycerol content were studied. ANOVAs indicated significant differences regarding chill coma recovery time, super-cooling point, and glycerol content across populations and sexes. The narrow-sense heritability (h2) estimates of cold resistance based on a parental half-sibling breeding design ranged from 0.39 to 0.53, and the h2 value was significantly higher in the Yangzhou population than in the Laibin population. Additive genetic variances were significantly different from zero for cold tolerance. The Yangzhou population of O. communa has a strong capability to quickly gain resistance to cold. We conclude that the O. communa beetle has a plasticity that can provide cold resistance in the changing climate conditions.

For tree species adapted to shallow soil environments, rooting strategies that efficiently explore rock fractures are important because soil water depletion occurs frequently. However, two questions: (a) to what extent shallow soil-adapted species rely on exploring rock fractures and (b) what outcomes result from drought stress, have rarely been tested. Therefore, based on the expectation that early development of roots into deep soil layers is at the cost of aboveground growth, seedlings of three tree species (Cyclobalanopsis glauca, Delavaya toxocarpa, and Acer cinnamomifolium) with distinct aboveground growth rates were selected from a typical shallow soil region. In a greenhouse experiment that mimics the basic features of shallow soil environments, 1-year-old seedlings were transplanted into simulated microcosms of shallow soil overlaying fractured bedrock. Root biomass allocation and leaf physiological activities, as well as leaf δ13C values were investigated and compared for two treatments: regular irrigation and repeated cycles of drought stress. Our results show that the three species differed in their rooting strategies in the context of encountering rock fractures, however, these strategies were not closely related to the aboveground growth rate. For the slowest-growing seedling, C. glauca, percentages of root mass in the fractures, as well as in the soil layer between soil and bedrock increased significantly under both treatments, indicating a specialized rooting strategy that facilitated the exploration of rock fractures. Early investment in deep root growth was likely critical to the establishment of this drought-vulnerable species. For the intermediate-growing, A. cinnamomifolium, percentages of root mass in the bedrock and interface soil layers were relatively low and exhibited no obvious change under either treatment. This limited need to explore rock fractures was compensated by a conservative water use strategy. For the fast-growing, D. toxocarpa, percentages of root mass in the bedrock and interface layers increased simultaneously under drought conditions, but not under irrigated conditions. This drought-induced rooting plasticity was associated with drought avoidance by this species. Although, root development might have been affected by the simulated microcosm, contrasting results among the three species indicated that efficient use of rock fractures is not a necessary or specialized strategy of shallow-soil adapted species. The establishment and persistence of these species relied on the mutual complementation between their species-specific rooting strategies and drought adaptations.

OBJECTIVE: To determine the capacity of dendritic cells (DCs) loaded with single or multiple-peptide mixtures of novel hepatitis C virus (HCV) epitopes to stimulate HCV-specific cytotoxic T lymphocyte (CTL) effector functions. METHODS: A bioinformatics approach was used to predict HLA-A2-restricted HCV-specific CTL epitopes, and the predicted peptides identified from this screen were synthesized. Subsequent IFN-γ ELISPOT analysis detected the stimulating function of these peptides in peripheral blood mononuclear cells (PBMCs) from both chronic and self-limited HCV infected subjects (subjects exhibiting spontaneous HCV clearance). Mature DCs, derived in vitro from CD14(+) monocytes harvested from the study subjects by incubation with appropriate cytokine cocktails, were loaded with novel peptide or epitope peptide mixtures and co-cultured with autologous T lymphocytes. Granzyme B (GrB) and IFN-γ ELISPOT analysis was used to test for epitope-specific CTL responses. T-cell-derived cytokines contained in the co-cultured supernatant were detected by flow cytometry. RESULTS: We identified 7 novel HLA-A2-restricted HCV-specific CTL epitopes that increased the frequency of IFN-γ-producing T cells compared to other epitopes, as assayed by measuring spot forming cells (SFCs). Two epitopes had the strongest stimulating capability in the self-limited subjects, one found in the E2 and one in the NS2 region of HCV; five epitopes had a strong stimulating capacity in both chronic and self-limited HCV infection, but were stronger in the self-limited subjects. They were distributed in E2, NS2, NS3, NS4, and NS5 regions of HCV, respectively. We also found that mDCs loaded with novel peptide mixtures could significantly increase GrB and IFN-γ SFCs as compared to single peptides, especially in chronic HCV infection subjects. Additionally, we found that DCs pulsed with multiple epitope peptide mixtures induced a Th1-biased immune response. CONCLUSIONS: Seven novel and strongly stimulating HLA-A2-restricted HCV-specific CTL epitopes were identified. Furthermore, DCs loaded with multiple-epitope peptide mixtures induced epitope-specific CTLs responses.