Causal relationship between gut microbiota, plasma metabolites, inflammatory cytokines and abdominal aortic aneurysm: a Mendelian randomization study.

BACKGROUND: Complex interconnections are evident among gut microbiota, circulating metabolites, inflammatory cytokines, and the pathogenesis of abdominal aortic aneurysms (AAA), with the causal dynamics yet to be comprehensively elucidated. The primary objective of this study was to elucidate the potential causal relationships involving gut microbiota-mediated plasma metabolites, inflammatory cytokines, and AAA. METHODS: We utilized data from genome-wide association studies predominantly comprising individuals of European ancestry, encompassing four major gut microbiota signatures, 233 plasma metabolite signatures (N = 136,016), 91 inflammatory cytokine signatures (N = 14,824), and AAA signatures (N = 1,458,875). Mendelian randomization (MR), employed in a two-sample format, was utilized as a tool to investigate the potential causal pathways from gut microbiota to the development of AAA. Additionally, a two-step MR approach was employed to dissect the impact of plasma metabolites and inflammatory cytokines on the relationship between gut microbiota and AAA and to ascertain the mediated fractions. RESULTS: Our findings indicate that five phylum or family-identical bacteria, 175 plasma metabolites, and seven inflammatory factors are causally associated with AAA. Among them, five bacterial species from the same phylum or family, identified from different GWAS data, were strongly associated with AAA. Of these, two exhibited negative causality and three exhibited positive causality. We found that the phylum Firmicutes and the families Oscillospiraceae might reduce the risk of AAA, whereas the families Prevotellaceae, Sutterellaceae, and Aminobacteriaceae might increase the risk of AAA. Further screening indicated that phylum Firmicutes id.1672 (GCST90017114) may confer a protective effect against AAA by reducing triglyceride levels in medium/small high-density lipoprotein (HDL). CONCLUSION: MR analysis has delineated a causal pathway from gut microbiota, through plasma circulating metabolites and inflammatory cytokines, to the pathogenesis of AAA. The role of intestinal flora and certain biomarkers may provide a reference for the diagnosis of AAA, and contribute to the prevention, diagnosis, and treatment of AAA disease.

Value of adjuvant chemotherapy for patients with pT2N0M0 non-small cell lung cancer receiving radical resection.

BACKGROUND: Associations between adjuvant chemotherapy (ACT) and the improvement in survival for patients with pT2N0M0 non-small cell lung cancer (NSCLC) who received R0 resection remain controversial. This study aimed to evaluate the value of ACT for patients with pT2N0M0 NSCLCs, and to identify the subgroups who could benefit from ACT. METHODS: Multivariable Cox proportional hazards regression models were used to estimate independent prognostic factors. High-risk factor (HRF) included visceral pleural invasion (VPI), lymphovascular invasion (LVI) and poor differentiation/undifferentiated tumors. RESULTS: Of the 899 patients, 277 (30.8%) patients received ACT. More younger patients (p < 0.001) and male patients (p = 0.007) received ACT. With the increase of pathological tumor size (p < 0.001) and the number of HRFs (p < 0.001), there was a significant rise in the proportion of patients receiving ACT. For all patients, ACT could not improve recurrence-free survival (RFS) (p = 0.672) and overall survival (OS) (p = 0.306). For patients with pathological stage IIA or radiological pure-solid tumors, ACT could significantly improve the OS (p = 0.011 and p = 0.037, respectively), and multivariate analysis revealed that ACT was an independent prognostic factor for patients with pathological stage IIA (p = 0.005). ACT could improve the OS significantly in patients with pathological stage IB pure-solid lung adenocarcinoma (LUAD) (p = 0.043). CONCLUSION: ACT was valuable for patients with pathological stage IIA (pT2bN0M0) and patients with radiological pure-solid LUAD of pathological stage IB. A combination of radiological features and pathological subtypes could be helpful when selecting patients with pT2N0M0 NSCLCs for ACT.

Identification of a coagulation-related signature correlated with immune infiltration and their prognostic implications in lung adenocarcinoma.

BACKGROUND: Lung adenocarcinoma (LUAD) is a fatal form of lung cancer with a poor prognosis. Coagulation system had been confirmed closely related to tumor progression and the hypercoagulable state encouraged the immune infiltration and development of tumor cells, leading to a poor prognosis in cancer patients. However, the use of the coagulation-related genes (CRGs) for prognosis in LUAD has yet to be determined. In this study, we constructed an immune-related signature (CRRS) and identified a potential coagulation-related biomarker (P2RX1). METHODS: We obtained a total of 209 CRGs based on two coagulation-related KEGG pathways, then developed the CRRS signature by using the TCGA-LUAD RNA-seq data via the procedure of LASSO-Cox regression, stepwise-Cox regression, univariate and multivariate Cox regression. Grouped by the CRRS, Kaplan-Meier survival curves and receiver operating characteristic curves were drawn for the training and validation sets, respectively. In addition, single-sample gene set enrichment analysis was exploited to explore immune infiltration level. Moreover, immunophenotypes and immunotherapy grouped by CRRS were further analyzed. RESULTS: We developed an immune-related signature (CRRS) composed of COL1A2, F2, PLAUR, C4BPA, and P2RX1 in LUAD. CRRS was an independent risk factor for overall survival and displayed stable and powerful performance. Additionally, CRRS possessed distinctly superior accuracy than traditional clinical variables and molecular features. Functional analysis indicated that the differentially high expressed genes in the low-risk group significantly enriched in T cell and B cell receptor signaling pathways. The low-risk group was sensitive to anti-PD-1/PD-L1 immunotherapy and displayed abundant immune infiltration and immune checkpoint gene expression. Finally, we identified an independent prognostic gene P2RX1. Low expression of P2RX1 associated with poor overall survival and decreased immune infiltration. CONCLUSIONS: Our study revealed a significant correlation between CRRS and immune infiltration. CRRS could serve as a promising tool to improve the clinical outcomes for individual LUAD patients.

PatchMix Augmentation to Identify Causal Features in Few-Shot Learning.

The task of Few-shot learning (FSL) aims to transfer the knowledge learned from base categories with sufficient labelled data to novel categories with scarce known information. It is currently an important research question and has great practical values in the real-world applications. Despite extensive previous efforts are made on few-shot learning tasks, we emphasize that most existing methods did not take into account the distributional shift caused by sample selection bias in the FSL scenario. Such a selection bias can induce spurious correlation between the semantic causal features, that are causally and semantically related to the class label, and the other non-causal features. Critically, the former ones should be invariant across changes in distributions, highly related to the classes of interest, and thus well generalizable to novel classes, while the latter ones are not stable to changes in the distribution. To resolve this problem, we propose a novel data augmentation strategy dubbed as PatchMix that can break this spurious dependency by replacing the patch-level information and supervision of the query images with random gallery images from different classes from the query ones. We theoretically show that such an augmentation mechanism, different from existing ones, is able to identify the causal features. To further make these features to be discriminative enough for classification, we propose Correlation-guided Reconstruction (CGR) and Hardness-Aware module for instance discrimination and easier discrimination between similar classes. Moreover, such a framework can be adapted to the unsupervised FSL scenario. The utility of our method is demonstrated on the state-of-the-art results consistently achieved on several benchmarks including miniImageNet, tieredImageNet, CIFAR-FS, CUB, Cars, Places and Plantae, in all settings of single-domain, cross-domain and unsupervised FSL. By studying the intra-variance property of learned features and visualizing the learned features, we further quantitatively and qualitatively show that such a promising result is due to the effectiveness in learning causal features.

Causal Association of Thyroid Signaling with C-Reactive Protein: A Bidirectional Mendelian Randomization.

Methods: Based on the latest genome-wide association study summary data, bidirectional two-sample Mendelian randomization (MR) was employed to detect the causal relationship and effect direction between TSH, fT4, and CRP. Furthermore, in view of obesity being an important risk factor of CVD, obesity trait waist-hip ratio (WHR) and body mass index (BMI) were treated as the research objects in MR analyses for exploring the causal effects of TSH and fT4 on them, respectively. Results: Genetically increased CRP was associated with increased TSH (ß = -0.02, P = 0.011) and with increased fT4 (ß = 0.043, P = 0.001), respectively, but there was no evidence that TSH or fT4 could affect CRP. In further analyses, genetically increased TSH was associated with decreased WHR (ß = -0.02, P = 3.99e - 4). Genetically increased WHR was associated with decreased fT4 (ß = -0.081, P = 0.002). Genetically increased BMI was associated with increased TSH (ß = 0.03, P = 0.028) and with decreased fT4 (ß = -0.078, P = 1.05e - 4). Causal associations of WHR and BMI with thyroid signaling were not supported by weighted median analysis in sensitivity analyses. Conclusion: TSH and fT4 were increased due to the higher genetically predicted CRP. WHR was decreased due to the higher genetically predicted TSH. These findings will provide reference for the prevention and treatment of inflammation and metabolic syndrome.

A Novel Hierarchical Clustering Approach for Joint Analysis of Multiple Phenotypes Uncovers Obesity Variants Based on ARIC.

Genome-wide association studies (GWASs) have successfully discovered numerous variants underlying various diseases. Generally, one-phenotype one-variant association study in GWASs is not efficient in identifying variants with weak effects, indicating that more signals have not been identified yet. Nowadays, jointly analyzing multiple phenotypes has been recognized as an important approach to elevate the statistical power for identifying weak genetic variants on complex diseases, shedding new light on potential biological mechanisms. Therefore, hierarchical clustering based on different methods for calculating correlation coefficients (HCDC) is developed to synchronously analyze multiple phenotypes in association studies. There are two steps involved in HCDC. First, a clustering approach based on the similarity matrix between two groups of phenotypes is applied to choose a representative phenotype in each cluster. Then, we use existing methods to estimate the genetic associations with the representative phenotypes rather than the individual phenotypes in every cluster. A variety of simulations are conducted to demonstrate the capacity of HCDC for boosting power. As a consequence, existing methods embedding HCDC are either more powerful or comparable with those of without embedding HCDC in most scenarios. Additionally, the application of obesity-related phenotypes from Atherosclerosis Risk in Communities via existing methods with HCDC uncovered several associated variants. Among these, UQCC1-rs1570004 is reported as a significant obesity signal for the first time, whose differential expression in subcutaneous fat, visceral fat, and muscle tissue is worthy of further functional studies.

Prognostic implication of tumor spread through air spaces in patients with pathologic N0 lung adenocarcinoma.

OBJECTIVES: Tumor spread through air spaces (STAS) has been identified as an invasive pattern in lung adenocarcinoma (ADC), but the prognostic implication of STAS has not been well studied in patients with pathologic N0 lung ADC. The purpose of this study was to evaluate the prognostic implication of STAS in pathologic N0 lung ADC patients after radical surgery. MATERIALS AND METHODS: Between January 2017 and December 2018, 796 patients with completely resected pathologic N0 lung ADC were reviewed. Pearson's chi-square test or Fisher exact test was used for comparing the relationship between STAS and clinicopathological features. The log-rank test and multivariate Cox regression models were used to explore prognostic factors. RESULTS: Among the 796 patients, STAS was positive in 201 patients (25.3%). The presence of STAS was significantly associated with patients with solid nodules (P < 0.001), micropapillary pattern-predominant adenocarcinoma/solid pattern-predominant adenocarcinoma (P < 0.001), larger tumor size (P < 0.001), visceral pleural invasion (P < 0.001) and lymphovascular invasion (P < 0.001). Multivariable analysis showed that STAS was an independent prognostic factor for recurrence-free survival (RFS) in pathologic N0 lung ADC patients (P = 0.014). For patients with acinar pattern-predominant adenocarcinoma (APA) / papillary pattern-predominant adenocarcinoma (PPA) / invasive mucinous adenocarcinoma (IMA) and patients who underwent lobectomy, STAS was an independent prognostic factor for RFS (P = 0.015, P = 0.011; respectively) and overall survival (OS) (P = 0.038, P = 0.020; respectively). CONCLUSION: In this study, STAS was an independent prognostic factor for RFS in pathologic N0 lung adenocarcinomas, and it was also an independent prognostic factor for RFS and OS in patients with APA/PPA/IMA and those who received lobectomy.

Clinicopathologic features and prognostic value of epidermal growth factor receptor mutation in patients with pT1a and pT1b invasive lung adenocarcinoma after surgical resection.

BACKGROUND: Previous studies have evaluated the prognostic value of epidermal growth factor receptor (EGFR) mutation in different subgroups of lung adenocarcinoma, but there remains controversial on this issue. We conduct this study aimed to reveal the prognostic value of EGFR mutation in patients with pT1a and pT1b invasive lung adenocarcinoma. METHODS: From August 2009 to February 2015, 338 patients with pT1a and pT1b invasive lung adenocarcinoma who underwent EGFR mutation analysis were enrolled into this study. According to clinicopathologic and radiologic characteristics, survival analysis was conducted in different subgroups using Kaplan-Meier methods and Cox regression models. RESULTS: EGFR mutation was detected in 216 (63.9%) patients. In the entire cohort, EGFR mutation was significantly frequent in female (P=0.011), never smoking (P=0.014) patients, patients with part-solid nodules (P=0.005) and patients with lepidic pattern-predominant adenocarcinoma (LPA)/acinar pattern-predominant adenocarcinoma (APA)/papillary pattern-predominant adenocarcinoma (PPA) (P=0.005). No difference in recurrence-free survival (RFS) was seen between patients harboring EGFR mutation and patients without EGFR mutation in the entire cohort (P=0.664) and the subgroup cohorts. Patients with EGFR mutation had a longer overall survival (OS) compared with patients without EGFR mutation in the entire cohort (P=0.005) and the subgroups of N0 stage cohort (P=0.013), N1-2 stage cohort (P=0.033), APA/PPA/invasive mucinous adenocarcinoma (IMA) cohort (P=0.011) and pT1b cohort (P=0.002). Tyrosine kinase inhibitors (TKIs) could significantly prolong the OS in patients with EGFR mutation after recurrence (P=0.04). CONCLUSIONS: EGFR mutation was not a risk factor for recurrence of patients with pT1a and pT1b invasive lung adenocarcinoma.

A Novel Method for Mendelian Randomization Analyses With Pleiotropy and Linkage Disequilibrium in Genetic Variants From Individual Data.

Mendelian randomization makes use of genetic variants as instrumental variables to eliminate the influence induced by unknown confounders on causal estimation in epidemiology studies. However, with the soaring genetic variants identified in genome-wide association studies, the pleiotropy, and linkage disequilibrium in genetic variants are unavoidable and may produce severe bias in causal inference. In this study, by modeling the pleiotropic effect as a normally distributed random effect, we propose a novel mixed-effects regression model-based method PLDMR, pleiotropy and linkage disequilibrium adaptive Mendelian randomization, which takes linkage disequilibrium into account and also corrects for the pleiotropic effect in causal effect estimation and statistical inference. We conduct voluminous simulation studies to evaluate the performance of the proposed and existing methods. Simulation results illustrate the validity and advantage of the novel method, especially in the case of linkage disequilibrium and directional pleiotropic effects, compared with other methods. In addition, by applying this novel method to the data on Atherosclerosis Risk in Communications Study, we conclude that body mass index has a significant causal effect on and thus might be a potential risk factor of systolic blood pressure. The novel method is implemented in R and the corresponding R code is provided for free download.

Optimized Intracellular Staining Reveals Heterogeneous Cytokine Production Ability of Murine and Human Hematopoietic Stem and Progenitor Cells.

Under stress conditions, hematopoietic stem and progenitor cells (HSPCs) can translate danger signals into a plethora of cytokine signals. These cytokines, or more precisely their combination, instruct HSPCs to modify the magnitude and composition of hematopoietic output in response to the threat, but investigations into the heterogeneous cytokine expression and regulatory mechanisms are hampered by the technical difficulty of measuring cytokine levels in HSPCs at the single-cell level. Here, we optimized a flow cytometry-based method for the simultaneous assessment of multiple intracellular cytokines in HSPCs. By selecting an optimal combination of cytokine restimulation reagents, protein transport inhibitors, and culture supplements, an optimized restimulation protocol for intracellular staining was developed. Using this method, we successfully examined expression levels of granulocyte/macrophage colony-stimulating factor (GM-CSF), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in murine and human HSPC subsets under steady-state or different stress conditions. Different cytokine expression patterns were observed, suggesting distinct regulatory modes of cytokine production dependent on the HSPC subset, cytokine, disease, organ, and species. Collectively, this technical advance may help to obtain a better understanding of the nature of HSPC heterogeneity on the basis of differential cytokine production.

Construction and Comprehensive Analysis of a ceRNA Network to Reveal Potential Novel Biomarkers for Triple-Negative Breast Cancer.

BACKGROUND: Triple-negative breast cancer (TNBC) is the most common and aggressive type of breast cancer with an unfavourable outcome worldwide. Novel therapeutic targets are urgently required to explore this malignancy. This study explored the ceRNA network and the important genes for predicting the therapeutic targets. METHODS: It identified the differentially expressed genes of mRNAs, lncRNAs and miRNAs between TNBC and non-TNBC samples in four cohorts (TCGA, GSE38959, GSE45827 and GSE65194) to explore the novel therapeutic targets for TNBC. Downstream analyses, including functional enrichment analysis, ceRNA network, protein-protein interaction and survival analysis, were then conducted by bioinformatics analysis. Finally, the potential core protein of the ceRNA network in TNBC was validated by immunohistochemistry. RESULTS: A total of 1,045 lncRNAs and 28 miRNAs were differentially expressed in the TCGA TNBC samples, and the intersections of 282 mRNAs (176 upregulations and 106 downregulations) between the GEO and TCGA databases were identified. A ceRNA network composed of 7 lncRNAs, 62 mRNAs, 12 miRNAs and 244 edges specific to TNBC was established. The functional assay showed dysregulated genes, and GO, DO and KEGG enrichment analysis were performed. Survival analysis showed that mRNA LIFR and lncRNA AC124312.3 were significantly correlated with the overall survival of patients with TNBC in the TCGA databases (P < 0.05). Finally, the LIFR protein was validated, and immunohistochemical results showed the upregulated expression of LIFR in TNBC tissues. CONCLUSION: Thus, our study presents an enhanced understanding of the ceRNA network in TNBC, where the key gene LIFR may be a new promising potential therapeutic target for patients with TNBC.

MASQC: Next Generation Sequencing Assists Third Generation Sequencing for Quality Control in N6-Methyladenine DNA Identification.

DNA N6-methyladenine (6mA) modification has been discovered as the most prevalent DNA modification in prokaryotes and eukaryotes, involving gene expression, DNA replication and repair, and host-pathogen interactions. Single-molecule real-time sequencing (SMRT-seq) can detect 6mA events in prokaryotic and eukaryotic genomes at the single-nucleotide level. However, there are no strict and economical quality control methods for high false-positive 6mA events in eukaryotic genomes. Therefore, by analyzing the distribution of 6mA in eukaryotic and prokaryotes, we proposed a method named MASQC (MeDIP-seq assists SMRT-seq for quality control in 6mA identification), which can identify 6mA events without doing the whole genome amplification (WGA) sequencing. The proposed MASQC method was assessed on two eukaryotic genomes and six bacterial genomes, our results demonstrate that MASQC performs well in quality control of false positive 6mA identification for both eukaryotic and prokaryotic genomes.

Artificial soil nutrient, aggregate stability and soil quality index of restored cut slopes along altitude gradient in southwest China.

In the subalpine mountainous region of southwest China, the artificial soil properties of restored cut slopes along the altitude gradient were studied, including available nitrogen (AN), available phosphorus (AP), available potassium (AK), urease (UR), sucrase (SC), protease (PR), catalase (CAT), texture, and aggregate stability of soil. Soil aggregatet stability by mean weight diameter (MWD), geometric mean diameter (GMD), structure failure rate (P), index of unstable aggregates (IUA) and area difference of dry and wet sieve cumulative distribution curve (ΔS) were measured and analyzed. It was found that available soil nutrients, UR, and CAT activities increased initially and then decreased, but the texture of soil was finer, and aggregate structure tended to be more stable along with an increase in the altitude gradient. The soil aggregate stability index that MWD, GMD, P, IUA, and ΔS indicated that the higher the altitude, the stability of soil aggregates was better. Principal component analysis (PCA) was used to determine the soil quality index (SQI). The obtained results from this study showed that the artificial soil quality of the cut slopes was better at a higher altitude compared to a lower altitude. These results provide a reference for the improvement of artificial soil properties of cut slopes and their quality in the future. It is necessary to pay attention to the soil quality management at a low-altitude area and reform the soil nutrients, enzyme activities, and soil structure for the restored cut slopes in the mountainous subalpine highway of southwest China.

Jatrorrhizine inhibits colorectal carcinoma proliferation and metastasis through Wnt/ß-catenin signaling pathway and epithelial-mesenchymal transition.

PURPOSE: Jatrorrhizine (JAT) is a natural protoberberine alkaloid, possesses detoxification, bactericidal and hypoglycemic activities. However, its anti-cancer mechanism is not clear. This study aimed to investigate the mechanism of JAT through which inhibits colorectal cancer in HCT-116 and HT-29 cells. METHODS: MTT assay and colony formation assay were used to check the cell proliferation ability. Cell apoptosis and cell cycle were measured by Hoechst 33342 staining and flow cytometry, respectively. Cell migration and invasion were detected by scratch wound healing assay and trans-well assay, respectively. Further, expression of related proteins was examined via Western blotting and the in vivo anti-cancer effect of JAT was confirmed by nude mice xenograft model. RESULTS: The research showed that JAT inhibited the proliferation of HCT-116 and HT-29 cells with IC50 values of 6.75±0.29 µM and 5.29±0.13 µM, respectively, for 72 hrs. It has also showed a time dependently, cell cycle arrested in S phase, promoted cell apoptosis and suppressed cell migration and invasion. In addition, JAT inhibited Wnt signaling pathway by reducing ß-catenin and increasing GSK-3ß expressions. Increased expression of E-cadherin, while decreased N-cadherin, indicating that JAT treatment suppressed the process of cell epithelial-mesenchymal transition (EMT). In HCT-116 nude mice xenograft model, JAT inhibited tumor growth and metastasis, and induced apoptosis of tumor cells. CONCLUSION: This study demonstrated that JAT efficiently inhibited colorectal cancer cells growth and metastasis, which provides a new point for clinical treatment of colorectal cancer.

The life span and influencing factors of metal mesh in artificial soil on railway rock-cut slopes in humid areas.

The stability of slope is strengthened by the metal mesh. The studies of the life span and influencing factors of metal mesh in the artificial soil in humid areas will guide ecological restoration of rock-cut slopes in Southwest China. Due to metal corrosion, the fixation function of the metal mesh could last for 10 years. The factors of soil contents, soil electrochemical properties and soil bacteria not only changed with the vegetation succession but also weakened the effect of the metal mesh on soil fixation for slope protection. The potential gradient, chloride ion content, sulfate ion content and water content were the main influencing factors for metal mesh corrosion during the vegetation restoration stage from 0 to 5 years, while the corrosion potential, potential gradient, chloride ion content, and water content were the main influential factors for metal mesh corrosion during the vegetation restoration stage after 11 years. At different vegetation restoration stages, the soil bacteria contained different dominant species, which had spatial heterogeneity, and the heterogeneity of the soil bacteria was another factor influencing the corrosion of the buried metal mesh. Meanwhile, the plant root as a soil fixation function strengthened with time, and 8 years later, the local woody plants gradually migrated to form a community dominated by multiple woody species. It is the first time that the life span of a metal mesh under the artificial soil of rock-cut slopes and factors affecting the different corrosion stages of the metal mesh in a humid area have been judged. The fixation function of plant root gradually replaces metal mesh, and main factors affecting the process include soil contents, soil electrochemical properties and soil bacteria. The research on corrosion factors of metal mesh under artificial soil for rock-cut slopes will contribute towards reducing the environmental risk of ecological restoration.

An efficient genomic signature ranking method for genomic island prediction from a single genome.

Genomic islands that are associated with microbial adaptations and carry genomic signatures different from that of the host, and thus many methods have been proposed to select the informative genomic signatures from a range of organisms and discriminate genomic islands from the rest of the genome in terms of these signature biases. However, they are of limited use when closely related genomes are unavailable. In the present work, we proposed a kurtosis-based ranking method to select the informative genomic signatures from a single genome. In simulations with alien fragments from artificial and real genomes, the proposed kurtosis-based ranking method efficiently selected the informative genomic signatures from a single genome, without annotated information of genomes or prior knowledge from other datasets. This understanding can be useful to design more powerful method for genomic island detection.

Vegetation and soil nutrient restoration of cut slopes using outside soil spray seeding in the plateau region of southwestern China.

Outside soil spray seeding (OSSS) is used widely for road cut revegetation, and the artificial soil used in OSSS can improve slope soil conditions and nutrients, and help promote plant growth and succession. Three different slopes was investigated to evaluate the effectiveness of OSSS for restoration, including a natural slope (NS), a cut slope without any artificial recovery treatment (CSW) and a cut slope treated with OSSS (CSO). The recovery of cut slopes was determined by evaluating a number of factors, including indices associated with plants on the slopes, soil enzyme activities (urease and sucrase), and soil nutrient content (soil organic matter (SOM), total phosphorous (TP), total potassium (TK), available nitrogen (AN), available phosphorous (AP), available potassium (AK), potassium (K+), calcium (Ca2+), magnesium (Mg2+), and sulphate (SO42-)). The results indicated that the vegetation and soil conditions differed between the three slopes. The Shannon-Wiener index (H), the Simpson index (D), and the Margalef index (R) values from the CSO and NS were lower than those of the CSW, whilst the Pielou index (E) value and vegetation canopy cover were higher for the CSO and NS than for the CSW. The content of SOM and AN in soil from the CSO was lower than in soil from the NS and CSW, and content of many nutrients were higher in soil from the CSO than in soil from the NS and CSW. This suggests that the restoration of vegetation and soil nutrients on the CSO was relatively successful. Our results indicated that the use of OSSS to restore cut slopes is effective in plateau areas. However, despite improvements in soil nutrient levels, there were still nutritional imbalances. Therefore, more attention should be paid to balancing nutrients in the later stage of OSSS implementation for the recovery of cut slopes at high altitudes.

Topographic aspect affects the vegetation restoration and artificial soil quality of rock-cut slopes restored by external-soil spray seeding.

External-soil spray seeding (ESSS), a technique of spraying artificial soil materials onto bare slopes for vegetation cover construction, has been widely used to restore rock-cut slopes. However, studies on the effect of the practical application of this technique on different topographic aspects have been rarely performed. In this study, two topographic aspects, namely, north-facing versus south-facing, were investigated under two railway lines, and two local natural slopes (north-facing versus south-facing) were selected as references. Vegetation and soil conditions, which are paramount aspects of ecological restoration assessment, were characterized in terms of the richness and diversity indices, vegetation canopy cover, basic soil physico-chemical properties, and structural characteristics of these slopes. Results showed that (1) the topographic aspect significantly affected the vegetation restoration and artificial soil quality of rock-cut slopes restored by ESSS; (2) the ecological restoration effect of north-facing slopes were better than that of south-facing slopes; and (3) the vegetation and soil conditions of natural slopes were better than those of rock-cut slopes. Therefore, additional scientific management measures should be implemented to promote the ecological restoration of rock-cut slopes, especially for south-facing slopes.

PTX-loaded three-layer PLGA/CS/ALG nanoparticle based on layer-by-layer method for cancer therapy.

Poly (lactic-co-glycolic acid) (PLGA) nanoparticles are an ideal paclitaxel (PTX)-carrying system due to its biocompatibility and biodegradability. But it possessed disadvantage of drug burst release. In this research, a layer-by-layer deposition of chitosan (CS) and sodium alginate (ALG) was applied to modify the PLGA nanoparticles. The surface charges and morphology of the PLGA, PLGA/CS and PLGA/CS/ALG particles was measured by capillary electrophoresis and SEM and TEM, respectively. The drug encapsulation and loading efficiency were confirmed by ultraviolet spectrophotometer. The nanoparticles were stable and exhibited controlled drug release performance, with good cytotoxicity to human lung carcinoma cells (HepG 2). Cumulatively, our research suggests that this kind of three-layer nanoparticle with LbL-coated shield has great properties to act as a novel drug-loaded system.

Synthesis and characterization of poly(lactic acid-co-glycolic acid) complex microspheres as drug carriers.

Poly(lactic-co-glycolic) acid (PLGA) is synthesized via melt polycondensation directly from lactic acid and glycolic acid with a feed molar ratio of 75/25. Bovine serum albumin, which is used as model protein, is entrapped into the poly(lactic-co-glycolic acid) microspheres with particle size of 260.9 ± 20.0 nm by the double emulsification method. Then it is the first report of producing more carboxyl groups by poly(lactic-co-glycolic acid) surface hydrolysis. The purpose is developing poly(lactic-co-glycolic acid) microspheres surface, which is modified with chitosan by chemical reaction between carboxyl groups and amine groups. The particle size and the positive zeta potential of the poly(lactic-co-glycolic acid)/chitosan microspheres are 388.2 ± 35.6 nm and 10.4 ± 2.9 mV, respectively. The drug loading ratio and encapsulation efficacy of poly(lactic-co-glycolic acid)/chitosan microspheres are 36.3% and 57.5%, which are higher than PLGA microspheres. Furthermore, the drug burst release of poly(lactic-co-glycolic acid)/chitosan microspheres at 10 h is decreased to 21.72% while the corresponding value of the poly(lactic-co-glycolic acid) microsphere is 64.56%. These results reveal that surface hydrolysis modification of poly(lactic-co-glycolic acid) is an efficient method to improve the negative potential and chemical reaction properties of the polymer. And furthermore, this study shows that chitosan-modified poly(lactic-co-glycolic acid) microspheres is a promising system for the controlled release of pharmaceutical proteins.