PacBio sequencing of human fecal samples uncovers the DNA methylation landscape of 22 673 gut phages.

Gut phages have an important impact on human health. Methylation plays key roles in DNA recognition, gene expression regulation and replication for phages. However, the DNA methylation landscape of gut phages is largely unknown. Here, with PacBio sequencing (2120×, 4785 Gb), we detected gut phage methylation landscape based on 22 673 gut phage genomes, and presented diverse methylation motifs and methylation differences in genomic elements. Moreover, the methylation rate of phages was associated with taxonomy and host, and N6-methyladenine methylation rate was higher in temperate phages than in virulent phages, suggesting an important role for methylation in phage-host interaction. In particular, 3543 (15.63%) phage genomes contained restriction-modification system, which could aid in evading clearance by the host. This study revealed the DNA methylation landscape of gut phage and its potential roles, which will advance the understanding of gut phage survival and human health.

Predict the prevalence and incidence of Parkinson's disease using fractal interpolation-LSTM model.

The investigation of the prediction of disease population is a noticeable exploration topic in the field of sciences. As a type of neurological disease, the incidence and prevalence of Parkinson's disease are still difficult to accurately study. In this paper, a method is proposed to forecast the number of incident cases (NumIn), incidence rate (InRa), the number of prevalent cases (NumPr), and prevalence rate (PrRa) of Parkinson's disease in ten countries selected. Using past data on the incidence rate, the number of prevalent cases, and the prevalence rate from 1990 to 2019, three types of fractal interpolations with different fractal dimensions are constructed for reconstructing the past data, where the vertical scaling factors are determined by the method proposed in this article. Then, the Long Short-Term Memory (LSTM) model is employed to forecast the values of NumIn, InRa, NumPr, and PrRa with Parkinson's disease in 2020. Meanwhile, the autoregressive integrated moving average model is used to predict the values compared with the LSTM model. The evaluation metrics employed for error calculation include the root mean square error and the coefficient of determination (R2). According to the proposed optimal criteria, the best predicted results are the average of three types of prediction values based on the LSTM model by analyzing and comparing eight predicted results.

Roseburia intestinalis generated butyrate boosts anti-PD-1 efficacy in colorectal cancer by activating cytotoxic CD8+ T cells.

OBJECTIVE: Roseburia intestinalis is a probiotic species that can suppress intestinal inflammation by producing metabolites. We aimed to study the role of R. intestinalis in colorectal tumourigenesis and immunotherapy. DESIGN: R. intestinalis abundance was evaluated in stools of patients with colorectal cancer (CRC) (n=444) and healthy controls (n=575). The effects of R. intestinalis were studied in ApcMin/+ or azoxymethane (AOM)-induced CRC mouse models, and in syngeneic mouse xenograft models of CT26 (microsatellite instability (MSI)-low) or MC38 (MSI-high). The change of immune landscape was evaluated by multicolour flow cytometry and immunohistochemistry staining. Metabolites were profiled by metabolomic profiling. RESULTS: R. intestinalis was significantly depleted in stools of patients with CRC compared with healthy controls. R. intestinalis administration significantly inhibited tumour formation in ApcMin/+ mice, which was confirmed in mice with AOM-induced CRC. R. intestinalis restored gut barrier function as indicated by improved intestinal permeability and enhanced expression of tight junction proteins. Butyrate was identified as the functional metabolite generated by R. intestinalis. R. intestinalis or butyrate suppressed tumour growth by inducing cytotoxic granzyme B+, interferon (IFN)-γ+ and tumour necrosis factor (TNF)-α+ CD8+ T cells in orthotopic mouse models of MC38 or CT26. R. intestinalis or butyrate also significantly improved antiprogrammed cell death protein 1 (anti-PD-1) efficacy in mice bearing MSI-low CT26 tumours. Mechanistically, butyrate directly bound to toll-like receptor 5 (TLR5) receptor on CD8+ T cells to induce its activity through activating nuclear factor kappa B (NF-κB) signalling. CONCLUSION: R. intestinalis protects against colorectal tumourigenesis by producing butyrate, which could also improve anti-PD-1 efficacy by inducing functional CD8+ T cells. R. intestinalis is a potential adjuvant to augment anti-PD-1 efficacy against CRC.

Uncovering 1058 Novel Human Enteric DNA Viruses Through Deep Long-Read Third-Generation Sequencing and Their Clinical Impact.

BACKGROUND & AIMS: Lack of viral reference genomes poses a challenge to virome study. We investigated human gut virome and its clinical implication by ultra-deep metagenomic sequencing. METHODS: We extracted sufficient viral DNA from human feces for ultra-deep PacBio sequencing (>10 µg) and Illumina sequencing (>1 µg). Upon de novo assembly and 6 stages of strict filtering, viral genomes were generated and validated in 3 cohorts of 2819 published fecal metagenomes. Diagnostic performance of assembled viruses for colorectal cancer were tested in a training cohort and 2 independent validation cohorts. Virus mapping ratio, evolutionary history, and virus status (lytic or temperate) were also examined. RESULTS: The mean amount of extracted viral DNA increased by 14-fold compared with previous protocols. We obtained PacBio long reads and Illumina short reads with 290-fold higher depth than previous studies. We assembled and validated 1178 contigs as complete viral genomes, of which 1058 were newly identified. Thirteen viral genomes (398-839 kb) that are longer than the largest bacteriophage found in humans (393 kb) were discovered. Phylogenetic tree was constructed based on Hidden Markov Models alignment scores of 4 conserved viral proteins. Incorporating our assembled genomes into the National Center for Biotechnology Information database improved the mapping ratio of published metagenomes ≤18 times. Lytic viruses (75.9% ± 12.2% of total) were predominantly present in our sample. A biomarker panel of 14 novel viruses could discriminate patients with colorectal cancer from controls with an area under the receiver operating characteristics curve of 0.87 in the training cohort, which was validated with areas under the receiver operating characteristics curve of 0.85 and 0.73 in 2 independent cohorts. CONCLUSIONS: We uncovered 1058 novel human gut viruses. These findings can contribute to clinical diagnosis, current viral reference genome, and future virome investigation.

Altered Mycobiota Signatures and Enriched Pathogenic Aspergillus rambellii Are Associated With Colorectal Cancer Based on Multicohort Fecal Metagenomic Analyses.

BACKGROUND & AIMS: The enteric mycobiota is a major component of the human gut microbiota, but its role in colorectal cancer (CRC) remains largely elusive. We conducted a meta-analysis to uncover the contribution of the fungal mycobiota to CRC. METHODS: We retrieved fecal metagenomic data sets from 7 previous publications and established an additional in-house cohort, totaling 1329 metagenomes (454 with CRC, 350 with adenoma, and 525 healthy individuals). Mycobiota composition and microbial interactions were analyzed. Candidate CRC-enriched fungal species (Aspergillus rambellii) was functionally validated in vitro and in vivo. RESULTS: Multicohort analysis revealed that the enteric mycobiota was altered in CRC. We identified fungi that were associated with patients with CRC or adenoma from multiple cohorts. Signature CRC-associated fungi included 6 enriched (A rambellii, Cordyceps sp. RAO-2017, Erysiphe pulchra, Moniliophthora perniciosa, Sphaerulina musiva, and Phytophthora capsici) and 1 depleted species (A kawachii). Co-occurrent interactions among CRC-enriched fungi became stronger in CRC compared with adenoma and healthy individuals. Moreover, we reported the transkingdom interactions between enteric fungi and bacteria in CRC progression, of which A rambellii was closely associated with CRC-enriched bacteria Fusobacterium nucleatum. A rambellii promoted CRC cell growth in vitro and tumor growth in xenograft mice. We further identified that combined fungal and bacterial biomarkers were more accurate than panels with pure bacterial species to discriminate patients with CRC from healthy individuals (the area under the curve relative change increased by 1.44%-10.60%). CONCLUSIONS: This study reveals enteric mycobiota signatures and pathogenic fungi in stages of colorectal tumorigenesis. Fecal fungi can be used, in addition to bacteria, for noninvasive diagnosis of patients with CRC.

Squalene epoxidase drives cancer cell proliferation and promotes gut dysbiosis to accelerate colorectal carcinogenesis.

OBJECTIVE: Aberrant lipid metabolism is a hallmark of colorectal cancer (CRC). Squalene epoxidase (SQLE), a rate-limiting enzyme in cholesterol biosynthesis, is upregulated in CRC. Here, we aim to determine oncogenic function of SQLE and its interplay with gut microbiota in promoting colorectal tumourigenesis. DESIGN: Paired adjacent normal tissues and CRC from two cohorts were analysed (n=202). Colon-specific Sqle transgenic (Sqle tg) mice were generated by crossing Rosa26-lsl-Sqle mice to Cdx2-Cre mice. Stools were collected for metagenomic and metabolomic analyses. RESULTS: SQLE messenger RNA and protein expression was upregulated in CRC (p<0.01) and predict poor survival of patients with CRC. SQLE promoted CRC cell proliferation by inducing cell cycle progression and suppressing apoptosis. In azoxymethane-induced CRC model, Sqle tg mice showed increased tumourigenesis compared with wild-type mice (p<0.01). Integrative metagenomic and metabolomic analyses unveiled gut dysbiosis in Sqle tg mice with enriched pathogenic bacteria, which was correlated to increased secondary bile acids. Consistent with detrimental effect of secondary bile acids, gut barrier function was impaired in Sqle tg mice, with reduced tight junction proteins Jam-c and occludin. Transplantation of Sqle tg mice stool to germ-free mice impaired gut barrier function and stimulated cell proliferation compared with control mice stool. Finally, we demonstrated that terbinafine, a SQLE inhibitor, could be repurposed for CRC by synergising with oxaliplatin and 5-fluorouracil to inhibit CRC growth. CONCLUSION: This study demonstrates that SQLE mediates oncogenesis via cell intrinsic effects and modulation of gut microbiota-metabolite axis. SQLE represents a therapeutic target and prognostic marker in CRC.

Cigarette smoke promotes colorectal cancer through modulation of gut microbiota and related metabolites.

OBJECTIVE: Cigarette smoking is a major risk factor for colorectal cancer (CRC). We aimed to investigate whether cigarette smoke promotes CRC by altering the gut microbiota and related metabolites. DESIGN: Azoxymethane-treated C57BL/6 mice were exposed to cigarette smoke or clean air 2 hours per day for 28 weeks. Shotgun metagenomic sequencing and liquid chromatography mass spectrometry were parallelly performed on mice stools to investigate alterations in microbiota and metabolites. Germ-free mice were transplanted with stools from smoke-exposed and smoke-free control mice. RESULTS: Mice exposed to cigarette smoke had significantly increased tumour incidence and cellular proliferation compared with smoke-free control mice. Gut microbial dysbiosis was observed in smoke-exposed mice with significant differential abundance of bacterial species including the enrichment of Eggerthella lenta and depletion of Parabacteroides distasonis and Lactobacillus spp. Metabolomic analysis showed increased bile acid metabolites, especially taurodeoxycholic acid (TDCA) in the colon of smoke-exposed mice. We found that E. lenta had the most positive correlation with TDCA in smoke-exposed mice. Moreover, smoke-exposed mice manifested enhanced oncogenic MAPK/ERK (mitogen-activated protein kinase/extracellular signal­regulated protein kinase 1/2) signalling (a downstream target of TDCA) and impaired gut barrier function. Furthermore, germ-free mice transplanted with stools from smoke-exposed mice (GF-AOMS) had increased colonocyte proliferation. Similarly, GF-AOMS showed increased abundances of gut E. lenta and TDCA, activated MAPK/ERK pathway and impaired gut barrier in colonic epithelium. CONCLUSION: The gut microbiota dysbiosis induced by cigarette smoke plays a protumourigenic role in CRC. The smoke-induced gut microbiota dysbiosis altered gut metabolites and impaired gut barrier function, which could activate oncogenic MAPK/ERK signalling in colonic epithelium.

Integrative metabolomic characterisation identifies altered portal vein serum metabolome contributing to human hepatocellular carcinoma.

OBJECTIVE: Altered metabolites are important for the tumourigenicity of hepatocellular carcinoma (HCC). We performed integrative metabolomics analysis of the metabolites changes in portal venous blood and in comparison with the metabolites changes in liver tissues and stool samples of HCC patients and healthy liver donors. DESIGN: Serum (portal and central vein), liver tissue (HCC tumour and adjacent non-tumour, normal liver) and stool samples were collected from 102 subjects (52 HCC patients and 50 healthy controls) in the discovery cohort; and 100 subjects (50 HCC patients and 50 healthy controls) in an independent validation cohort. Untargeted metabolomic profiling was performed using high-performance liquid chromatography-mass spectrometry. The function of candidate metabolites was validated in hepatocyte cell lines. RESULTS: Detailed metabolomic evaluation showed distinct clusters of metabolites in serum, liver tissue and stool samples from patients with HCC and control individuals (p<0.001). HCC patients had significantly higher levels of portal vein serum and HCC tissue metabolites of DL-3-phenyllactic acid, L-tryptophan, glycocholic acid and 1-methylnicotinamide than healthy controls, which were associated with impaired liver function and poor survival. On the other hand, HCC patients had lower levels of linoleic acid and phenol in portal vein and stool samples than healthy controls. Linoleic acid and phenol significantly inhibited HCC proliferation, inferring their anti-HCC function as protective metabolites. CONCLUSIONS: The integrative metabolome analysis of serum, tissue and stool metabolites revealed unreported metabolic alterations in HCC patients. In portal vein, we identified elevated and depleted metabolites signifying that they might play a role in HCC development.

Transmission Line Object Detection Method Based on Contextual Information Enhancement and Joint Heterogeneous Representation.

Transmission line inspection plays an important role in maintaining power security. In the object detection of the transmission line, the large-scale gap of the fittings is still a main and negative factor in affecting the detection accuracy. In this study, an optimized method is proposed based on the contextual information enhancement (CIE) and joint heterogeneous representation (JHR). In the high-resolution feature extraction layer of the Swin transformer, the convolution is added in the part of the self-attention calculation, which can enhance the contextual information features and improve the feature extraction ability for small objects. Moreover, in the detection head, the joint heterogeneous representations of different detection methods are combined to enhance the features of classification and localization tasks, which can improve the detection accuracy of small objects. The experimental results show that this optimized method has a good detection performance on the small-sized and obscured objects in the transmission line. The total mAP (mean average precision) of the detected objects by this optimized method is increased by 5.8%, and in particular, the AP of the normal pin is increased by 18.6%. The improvement of the accuracy of the transmission line object detection method lays a foundation for further real-time inspection.

PVformer: Pedestrian and Vehicle Detection Algorithm Based on Swin Transformer in Rainy Scenes.

Pedestrian and vehicle detection plays a key role in the safe driving of autonomous vehicles. Although transformer-based object detection algorithms have made great progress, the accuracy of detection in rainy scenarios is still challenging. Based on the Swin Transformer, this paper proposes an end-to-end pedestrian and vehicle detection algorithm (PVformer) with deraining module, which improves the image quality and detection accuracy in rainy scenes. Based on Transformer blocks, a four-branch feature mapping model was introduced to achieve deraining from a single image, thereby mitigating the influence of rain streak occlusion on the detector performance. According to the trouble of small object detection only by visual transformer, we designed a local enhancement perception block based on CNN and Transformer. In addition, the deraining module and the detection module were combined to train the PVformer model through transfer learning. The experimental results show that the algorithm performed well on rainy days and significantly improved the accuracy of pedestrian and vehicle detection.

Assessment of GF3 Full-Polarimetric SAR Data for Dryland Crop Classification with Different Polarimetric Decomposition Methods.

Crop classification is one of the most important agricultural applications of remote sensing. Many studies have investigated crop classification using SAR data, while few studies have focused on the classification of dryland crops by the new Gaofen-3 (GF3) SAR data. In this paper, taking Hengshui city as the study area, the performance of the Freeman-Durden, Sato4, Singh4 and multi-component decomposition methods for dryland crop type classification applications are evaluated, and the potential of full-polarimetric GF3 data in dryland crop type classification are also investigated. The results show that the multi-component decomposition method produces the most accurate overall classifications (88.37%). Compared with the typical polarization decomposition techniques, the accuracy of the classification results using the new decomposition method is improved. In addition, the Freeman method generally yields the third-most accurate results, and the Sato4 (87.40%) and Singh4 (87.34%) methods yield secondary results. The overall classification accuracy of the GF3 data is very positive. These results demonstrate the great promising potential of GF3 SAR data for dryland crop monitoring applications.

Efficacy and safety of laparoscopic splenectomy for hypersplenism secondary to portal hypertension after transjugular intrahepatic portosystemic shunt.

BACKGROUND: Laparoscopic splenectomy (LS) being used after Transjugular intrahepatic portosystemic shunt (TIPS) has not been reported. This report aims to explore the feasibility, safety, and potential efficacy of LS after TIPS hypersplenism secondary to portal hypertension (PHT). METHODS: We retrospectively reviewed a series of six patients who underwent LS after TIPS for hypersplenism secondary to PHT between 2014 and 2020. The perioperative data and patients' clinical outcomes were recorded. RESULTS: LS was successfully performed in all patients. Hypersplenism was corrected after LS in all six patients. Postoperative prothrombin time, prothrombin activity, international normalized ratio, and total bilirubin showed a trend toward improvement. The preoperative and 1-month postoperative albumin and activated partial thromboplastin levels showed no significant difference. Plasma ammonia level and thromboelastography indicators were ameliorated in two limited recorded patients. No postoperative complications such as subphrenic abscess, portal vein thrombosis, variceal bleeding, hepatic encephalopathy, and liver failure occurred during the 1-month follow-up period. CONCLUSION: LS following TIPS is feasible, safe, and beneficial for patients with hypersplenism secondary to PHT. The following LS not only corrects the hypersplenism, but also has the potential to improve liver function.

Dryland Crop Classification Combining Multitype Features and Multitemporal Quad-Polarimetric RADARSAT-2 Imagery in Hebei Plain, China.

The accuracy of dryland crop classification using satellite-based synthetic aperture radar (SAR) data is often unsatisfactory owing to the similar dielectric properties that exist between the crops and their surroundings. The main objective of this study was to improve the accuracy of dryland crop (maize and cotton) classification by combining multitype features and multitemporal polarimetric SAR (PolSAR) images in Hebei plain, China. Three quad-polarimetric RADARSAT-2 scenes were acquired between July and September 2018, from which 117 features were extracted using the Cloude-Pottier, Freeman-Durden, Yamaguchi, and multiple-component polarization decomposition methods, together with two polarization matrices (i.e., the coherency matrix and the covariance matrix). Random forest (RF) and support vector machine (SVM) algorithms were used for classification of dryland crops and other land-cover types in this study. The accuracy of dryland crop classification using various single features and their combinations was compared for different imagery acquisition dates, and the performance of the two algorithms was evaluated quantitatively. The importance of all investigated features was assessed using the RF algorithm to optimize the features used and the imagery acquisition date for dryland crop classification. Results showed that the accuracy of dryland crop classification increases with evolution of the phenological period. In comparison with SVM, the RF algorithm showed better performance for dryland crop classification when using full polarimetric RADARSAT-2 data. Dryland crop classification accuracy was not improved substantially when using only backscattering intensity features or polarization decomposition parameters extracted from a single-date image. Satisfactory classification accuracy was achieved using 11 optimized features (derived from the Cloude-Pottier decomposition and the coherency matrix) from 2 RADARSAT-2 images (acquisition dates corresponding to the middle and late stages of dryland crop growth). This study provides an important reference for timely and accurate classification of dryland crop in Hebei plain, China.

Challenges of the establishment of rubber-based agroforestry systems: Decreases in the diversity and abundance of ground arthropods.

The global land area devoted to rubber plantations has now reached 13 million hectares, and the further expansion of these rubber plantations at the expense of tropical forests will have significant adverse effects on the ecological environment. Rubber-based agroforestry systems are considered a preferable approach for ameliorating the ecological environment. Many researchers have focused on the positive effects of rubber-based agroforestry systems on the ecological environment, while ignoring the risks involved in the establishment of rubber-based agroforestry systems. The present study investigated the effects of different-aged rubber-based agroforestry systems on the abundance and diversity of ground arthropods. It has been observed that the abundance and taxon richness of ground arthropods generally showed no difference when comparing young and mature rubber plantations. The rubber-based agroforestry systems significantly decreased the understory vegetation species, along with the abundance and taxon richness of ground arthropods compared to the same aged-rubber monoculture plantations. In addition, the change in the abundance and taxon richness of ground arthropods was greatly affected by the understory vegetation species and soil temperature. The abundance and taxon richness of ground arthropods decreased with the decrease in number of species of understory vegetation. The study results indicate that the establishment of rubber-based agroforestry systems have adversely affected the abundance and richness of ground arthropods to an extant greater than expected. Therefore, single, large rubber-based agroforestry systems are not recommended, and the intercropping of rubber and rubber-based agroforestry systems must be designed to promote the migration of ground arthropods between different systems.

Contrasting ecosystem CO2 fluxes of inland and coastal wetlands: a meta-analysis of eddy covariance data.

Wetlands play an important role in regulating the atmospheric carbon dioxide (CO2 ) concentrations and thus affecting the climate. However, there is still lack of quantitative evaluation of such a role across different wetland types, especially at the global scale. Here, we conducted a meta-analysis to compare ecosystem CO2 fluxes among various types of wetlands using a global database compiled from the literature. This database consists of 143 site-years of eddy covariance data from 22 inland wetland and 21 coastal wetland sites across the globe. Coastal wetlands had higher annual gross primary productivity (GPP), ecosystem respiration (Re ), and net ecosystem productivity (NEP) than inland wetlands. On a per unit area basis, coastal wetlands provided large CO2 sinks, while inland wetlands provided small CO2 sinks or were nearly CO2 neutral. The annual CO2 sink strength was 93.15 and 208.37 g C m-2 for inland and coastal wetlands, respectively. Annual CO2 fluxes were mainly regulated by mean annual temperature (MAT) and mean annual precipitation (MAP). For coastal and inland wetlands combined, MAT and MAP explained 71%, 54%, and 57% of the variations in GPP, Re , and NEP, respectively. The CO2 fluxes of wetlands were also related to leaf area index (LAI). The CO2 fluxes also varied with water table depth (WTD), although the effects of WTD were not statistically significant. NEP was jointly determined by GPP and Re for both inland and coastal wetlands. However, the NEP/Re and NEP/GPP ratios exhibited little variability for inland wetlands and decreased for coastal wetlands with increasing latitude. The contrasting of CO2 fluxes between inland and coastal wetlands globally can improve our understanding of the roles of wetlands in the global C cycle. Our results also have implications for informing wetland management and climate change policymaking, for example, the efforts being made by international organizations and enterprises to restore coastal wetlands for enhancing blue carbon sinks.

Overexpression of microRNA-30a-5p inhibits liver cancer cell proliferation and induces apoptosis by targeting MTDH/PTEN/AKT pathway.

Increasing results suggest microRNAs (miRNAs) could function important roles in malignant tumor progression. miR-30a-5p is downregulated in variety of cancers and acts as a cancer suppressing gene. The functions and molecular mechanisms of miRNA-30a-5p in hepatocellular carcinoma (HCC) remain unclear. In the present study, quantitative real-time PCR (qRT-PCR) was used to detect miR-30a-5p expression in 16 pairs of HCC and their adjacent non-cancerous tissues and HCC cell lines. By overexpression of miRNA-30a-5p, CCK-8 and colon formation assay were used to evaluate cell growth and flow cytometry to evaluate cell apoptosis. Western blot was used to test protein expression. And potential mechanisms were analyzed with luciferase activity assay. In vivo HepG2 tumor growth was observed with nude mice. Our results showed that miR-30a-5p expression in HCC tissues was significantly lower compared to adjacent non-cancerous liver tissues, and lower miR-30a-5p expression was also observed in HCC cell lines compared to normal liver cell. Luciferase assay showed that metadherin (MTDH) mRNA was a direct target of miR-30a-5p. A significant reverse correlation between miR-30a-5p and MTDH in liver cancer tissues was observed. miR-30a-5p overexpression in HCC cells significantly inhibited cell proliferation, colon formation, and induced apoptosis while MTDH overexpression reversed growth inhibition and apoptosis induction of miRNA-30a-5p in HCC cells. miRNA-30a-5p upregulated phosphatase and tensin homolog (PTEN) protein expression and thus inhibited AKT activating by targeting MTDH. miRNA-30a-5p also significantly inhibited HepG2 tumor growth in vivo. Our results suggest that underexpression of miR-30a-5p might function as a tumor suppressing miRNA by directly targeting MTDH in HCC and is therefore a potential candidate biomarker for HCC targeting therapy.

High expression of metadherin correlates with malignant pathological features and poor prognostic significance in papillary thyroid carcinoma.

BACKGROUND: Metadherin (MTDH) protein, also called astrocyte elevated gene-1 (AEG-1) is over expressed in a variety of malignant tumours, and is closely related to tumour invasion and the poor prognosis. OBJECTIVE: This study tries to explore the clinical pathological significance of MTDH expression in a large cohort of patients with PTC. DESIGN AND PATIENTS: Immunohistochemistry was used to detect MTDH expression in 156 cases of PTC, 6 cases of anaplastic thyroid carcinoma (ATC), 10 cases of multinodular goitre (MNG) and 10 cases of thyroid adenoma tissues who received a thyroid operation between June 2003 and July 2008. MEASUREMENTS: Clinical pathological data of 156 cases of PTC were analysed according to MTDH expression. The Kaplan-Meier method was used to plot survival curves and log-rank test to compare the postoperative survival results. The prognostic meaning of MTDH expression in PTC was evaluated by Cox regression analysis. RESULTS: The positive expression rates of MTDH in PTC and ATC tissues were 37·2% (58/156) and 50% (3/6), respectively, and MTDH positive expression rates were both 10% (1/10) in MNG and thyroid adenoma tissues. High MTDH expression in PTC was associated with larger tumour size (P = 0·030), high rates of lymph node (P = 0·041) and distant metastasis (P = 0·028), but no relation with the patient age, gender, tumour multicenter, extrathyroid invasion and tumour grade. High MTDH expression was associated with recurrence-free survival (RFS) and disease-specific survival rate (DSS) (P = 0·014, P = 0·001, respectively). Cox regression analysis showed that high MTDH expression was independent prognostic indicators for RFS and DSS in patients with PTC (P = 0·023 and P = 0·035, respectively). CONCLUSION: High MTDH expression in PTC might play an important role in tumour growth and metastasis, and targeting MTDH treatment might have potential therapeutic value for patients with PTC.

Lentiviral-Mediated Short Hairpin RNA Knockdown of MTDH Inhibits Cell Growth and Induces Apoptosis by Regulating the PTEN/AKT Pathway in Hepatocellular Carcinoma.

The activation of oncogenes and the loss of tumor suppressor genes are believed to play critical roles in the pathogenesis of human hepatocellular carcinoma (HCC). Metaherin(MTDH), also called astrocyte elevated gene-1 (AEG-1), is frequently amplified in a variety of cancers, but the roles of MTDH with regard to growth and apoptosis in HCC have not yet been studied. In the present study, we first analyzed the expression of MTDH in HCC samples. We found that MTDH protein levels are higher in most HCC cancerous tissues compared with their matched adjacent non-tumor tissues. Additionally, the MTDH mRNA was also higher inHCC tissues compared to their matched adjacent non-tumor tissues. Knockdown of the endogenous MTDH using small interfering RNA further showed that deficiency of MTDH suppressed cell growth and caused apoptosis in HCC cells. Knockdown MTDH promoted PTEN and p53 expression in HCC cells and inhibited AKT phosphorylation. Knockdown MTDH also inhibited tumor growth in vivo. All these results indicated that MTDH protein levels in most HCC tissues are higher than non-tumor tissues, and knockdown of MTDH inhibited growth and induced apoptosis in HCC cells through the activation of PTEN.Therefore, MTDH might be an effective targeted therapy gene for HCC.

[Effects of hypoxia inducible factor-2α on promoting angiogenesis of residual hepatocellular carcinoma after high-intensity focused ultrasound ablation].

OBJECTIVE: To investigate the dynamic features of angiogenesis in residual tumors after high intensity focused ultrasound (HIFU),and to determine the temporal effect and mechanism of hypoxia inducible factor-2 alpha (HIF-2a) in the angiogenic process of residual tumors. METHODS: Xenograft tumors of HepG2 cells were generated by subcutaneously inoculating athymic BALB/c nu/nu mice with the hepatoma cells.About 30 days after inoculation,all mice (except in the control group) were treated by HIFU and assigned randomly to the following 7 groups according to various time intervals post-treatment:1st,3rd,5th day and 1st,2nd,3rd,4th week when the residual tumor tissues were obtained from the experimental groups.Protein levels of HIF-2a and vascular growth factor A (VEGF-A) were quantified by immunohistochemistry and western blotting,and mRNA levels were measured by (real-time quantitative) qPCR. Microvascular density (MVD) was calculated by counting the CD31-positive vascular endothelial cells identified by means of an immunohistochemical staining method. RESULTS: Compared with results from the control group,the protein and mRNA levels of HIF-2a expression reached the highest level in the experimental mice at the 2nd week (P=0.000 and P < 0.01 respectively),and were decreased thereafter(3rd week and 4th week, P=0.000 and P < 0.05).VEGF-A expression in the residual tumor tissues group that received HIFU was significantly decreased,compared with the control group,at all time points uPto 1 week (all P=0.000 and P < 0.01),but the levels increased compared to controls in the 2nd through 4th week (all P=0.000, P < 0.05). Similar results were obtained for MVD. CONCLUSION: HIFU treatment can inhibit angiogenesis in residual hepatoma tissues in the short-term (1 to 2 weeks post-treatment) in mice with hepatocellular carcinoma,but can promote angiogenesis overtime (2 to 4 weeks post-treatment); the angiogenic process may involve the HIF-2α/VEGFA pathway.

Improved soil moisture, nutrients, and economic benefits using plastic mulchs in balsa-based agroforestry systems.

The manufacture of wind turbine blades generally uses balsa wood as the base materials, and it is crucial to explore new regions for cultivating balsa trees to achieve carbon neutrality in the future. Xishuangbanna may be China's only area with a tropical climate suitable for the large-scale planting of balsa trees. The present study investigated the key soil elements influencing the growth of balsa plantations and the effects of different cultivation practices on soil environments and economic benefits in Xishuangbanna, China. We found that the height of balsa stems after growing 4 years reached 5.8 m; the increment of diameter at breast height (DBH) reached 27.7 cm and volume of balsa stems reached 196.0 m3 ha-1 in Xishuangbanna of China. It is of the utmost importance to improve the contents of soil exchangeable magnesium (Mg) and available phosphorus (P) for the growth of balsa trees, and exchangeable aluminium (Al) inhibited the growth of balsa trees. The practice of plastic film mulching not only improved soil moisture in the 40‒100-cm soil layer in the dry season and in the 0-60-cm soil layer in the rainy season but also enhanced soil nitrate nitrogen when compared with no plastic-mulching practice in balsa plantations. The comprehensive economic benefits of balsa/coriander/ginger/taro plantations were significantly improved by implementing plastic film mulching, as compared to balsa plantations. We conclude that balsa tree can be cultivated in Xishuangbanna, China, and its successful cultivation provides opportunities for China's wind power development.