Papillary thyroid carcinoma with clear cell renal cell carcinoma metastasized to the thyroid gland: A case report.

Metastasis of clear cell renal cell carcinoma (ccRCC) to the thyroid gland is rare, and simultaneous occurrence of ccRCC and papillary thyroid carcinoma (PTC) is even rarer. Due to the occult nature of the disease, the clinical diagnosis is difficult. In the case of multiple tumors, the possibility of thyroid metastasis should not be ignored during the clinical diagnosis and treatment of PTC. The present study reported a case with initial diagnosis of PTC and accidental discovery of thyroid metastasis of ccRCC. This case study aims to improve the understanding of occult thyroid metastasis, providing a reference for its clinical diagnosis and treatment. Accordingly, misdiagnosis and missed diagnosis of this disease may be reduced and the survival rate and the life quality of patients can be improved.

Spatial Distribution of Pinus koraiensis Trees and Community-Level Spatial Associations in Broad-Leaved Korean Pine Mixed Forests in Northeastern China.

Investigating the spatial distributions and associations of tree populations provides better insights into the dynamics and processes that shape the forest community. Korean pine (Pinus koraiensis) is one of the most important tree species in broad-leaved Korean pine mixed forests (BKMFs), and little is known about the spatial point patterns of and associations between Korean pine and community-level woody species groups such as coniferous and deciduous trees in different developmental stages. This study investigated the spatial patterns of Korean pine (KP) trees and then analyzed how the spatial associations between KP trees and other tree species at the community level vary in different BKMFs. Extensive data collected from five relatively large sample plots, covering a substantial area within the natural distribution range of KP in northeastern China, were utilized. Uni- and bivariate pair correlation functions and mark correlation functions were applied to analyze spatial distribution patterns and spatial associations. The DBH (diameter at breast height) histogram of KP trees in northeastern China revealed that the regeneration process was very poor in the Changbai Mountain (CBS) plot, while the other four plots exhibited moderate or expanding population structures. KP trees were significantly aggregated at scales up to 10 m under the HPP null model, and the aggregation scales decreased with the increase in size classes. Positive or negative spatial associations were observed among different life stages of KP trees in different plots. The life history stages of the coniferous tree group showed positive spatial associations with KP saplings and juvenile trees at small scales, and spatial independence or negative correlations with larger KP trees at greater scales. All broad-leaved tree groups (canopy, middle, and understory layers) exhibited only slightly positive associations with KP trees at small scales, and dominant negative associations were observed at most scales. Our results demonstrate that mature KP trees have strong importance in the spatial patterns of KP populations, and site heterogeneity, limited seed dispersal, and interspecific competition characterize the spatial patterns of KP trees and community-level spatial associations with respect to KP trees, which can serve as a theoretical basis for the management and restoration of BKMFs in northeastern China.

Saikosaponin-D induces the pyroptosis of lung cancer by increasing ROS and activating the NF-κB/NLRP3/caspase-1/GSDMD pathway.

Saikosaponin-D (SSD), an active ingredient in Bupleurum chinense, exerts anticancer effects in various cancers by inhibiting cancer proliferation and inducing apoptosis. However, whether SSD can induce other forms of cell death is unknown. The current study aims to demonstrate that SSD can induce pyroptosis in non-small-cell lung cancer. In this study, HCC827 and A549 non-small-cell lung cancer cells were treated with different concentrations of SSD for 1.5 h. HE and TUNEL staining were used to verify cell damage caused by SSD. Immunofluorescence and western blotting were performed to verify the effect of SSD on the NF-κB/NLRP3/caspase-1/gasdermin D (GSDMD) pathway. Changes in inflammatory factors were detected by ELISAs. Finally, the reactive oxygen species (ROS) scavenger N-acetylcysteine (NAC) was introduced to verify that SSD induces pyroptosis through the ROS/NF-κB pathway. The results of the HE and TUNEL staining showed that SSD resulted in balloon-like swelling of NSCLC cells accompanied by increased DNA damage. Immunofluorescence and western blot assays confirmed that SSD treatment activated the NLRP3/caspase-1/GSDMD pathway, stimulated an increase in ROS levels and activated NF-κB in lung cancer cells. The ROS scavenger N-acetylcysteine significantly attenuated SSD-induced NF-κB/NLRP3/caspase-1/GSDMD pathway activation and inhibited the release of the inflammatory cytokines IL-1ß and IL-18. In conclusion, SSD induced lung cancer cell pyroptosis by inducing ROS accumulation and activating the NF-κB/NLRP3/caspase-1/GSDMD pathway. These experiments lay the foundation for the application of SSD in the treatment of non-small-cell lung cancer and regulation of the lung cancer immune microenvironment.

Silencing MARCH2 Inhibits the Invasion, Migration, and EMT Transformation of Human Colorectal Cancer SW480 Cells.

Aim: This study aimed to investigate the role of MARCH2 (membrane-associated RING-CH2) in the progression, invasion, and migration of colorectal cancer (CRC). Methods: In this study, the expression levels of MARCH2 and E-cadherin in CRC tissues were detected by immunohistochemistry through retrospective study, and their correlation was analyzed. After silencing the MARCH2 gene using SiRNA MARCH2-1/-2, the invasion and migration abilities of SW480 cells were detected using Transwell and Scratch assay, respectively. Quantitative real-time PCR (qRT-PCR) and Western blotting assays were performed to detect the expression levels of epithelial-mesenchymal transition (EMT) related markers. Results: As compared to adjacent tissues, the MARCH2 expression level was significantly overexpressed in the CRC tissues, and correlated with tumor size, pathological grade, lymph node metastasis, and survival time. MARCH2 was negatively correlated with E-cadherin. MARCH2 silencing significantly restrained the invasion and migration abilities of SW480 cells in vitro. Meanwhile, the MARCH2 silencing also upregulated the mRNA and protein expression levels of E-cadherin and downregulated those of Vimentin. Conclusions: The high expression of MARCH2 was unfavorable for patients' survival. Thus, MARCH2 might be an independent predictor for CRC patients, affecting the invasion and metastasis of CRC through EMT.

Soil water content and nitrogen differentially correlate with multidimensional leaf traits of two temperate broadleaf species.

The variation and correlation of leaf economics and vein traits are crucial for predicting plant ecological strategies under different environmental changes. However, correlations between these two suites of traits and abiotic factors such as soil water and nitrogen content remain ambiguous. We measured leaf economics and vein traits as well as soil water and nitrogen content for two different shade-tolerant species (Betula platyphylla and Acer mono) in four mixed broadleaved-Korean pine (Pinus koraiensis) forests along a latitudinal gradient in Northeast China. We found that leaf economics traits and vein traits were decoupled in shade-intolerant species, Betula platphylla, but significantly coupled in a shade-tolerant species, A. mono. We found stronger correlations among leaf traits in the shade tolerant species than in the shade intolerant species. Furthermore, leaf economic traits were positively correlated with the soil water gradient for both species, whereas vein traits were positively correlated with soil water gradient for the shade intolerant species but negatively correlated in the shade tolerant species. Although economic traits were positively correlated with soil nitrogen gradient in shade intolerant species but not correlated in shade tolerant species, vein traits were negatively correlated with soil nitrogen gradient in shade tolerant species but not correlated in shade intolerant species. Our study provides evidence for distinct correlations between leaf economics and vein traits and local abiotic factors of species differing in light demands. We recommend that the ecological significance of shade tolerance be considered for species when evaluating ecosystem functions and predicting plant responses to environmental changes.

Long noncoding RNA CLAN promotes lymphangiogenesis in the colorectal carcinoma.

Metastasis is the main cause of colorectal cancer (CRC)-related death and lymph node plays a vital role in this process. Long noncoding RNAs (lncRNAs) are emerging as an important factor of biological progress in cancers. However, lncRNAs related to CRC metastasis was rarely reported.CLAN expression data of tumor tissues and normal tissues were obtained from GEPIA database and 23 paired tumor and normal samples of patients. CLAN expression of 85 patients was carried out with RNA extracted from FFPE samples and quantified with qRT-PCR. Patients' clinical features were collected from department of Pathology of the Affiliated Hospital of Southwest Medical University. Immunohistochemistry staining was used to detect the metastasis-related proteins.CLAN was highly expressed in tumor tissues. And the expression level was not correlated with age, gender, differentiation, and location of CRC patients. Also, CLAN expression did not correlated with budding, LVI, and TILs. However, CLAN expression was strongly associated with lymph node metastasis and higher TNM stage. CLAN changed the lymphatic vessel density by promoting lymphangiogenesis but CLAN did not affect the blood vessel density.CLAN was a unique lncRNA that promoted lymphangiogenesis to accelerate CRC metastasis. CLAN might play a unique role in tumor early dissemination through lymphatic vessel.

miR-576-5p Promotes the Proliferation of Papillary Thyroid Carcinoma through the MAPK4-AKT Pathway.

Background: MicroRNA-576-5p (miR-576-5p) plays an important role in different human cancers. However, the biological function of miR-576-5p in papillary thyroid carcinoma (PTC) is still unclear. In this study, we explored the function and specific role of miR-576-5p in PTC. Methods: Expression levels of miR-576-5p in PTC patient tissues and cell lines were determined by reverse transcription-quantitative polymerase chain reaction (qRT‒PCR). Cell counting using cell counting kit-8 (CCK-8), wound healing, and Transwell assays were performed to evaluate the effect of miR-576-5p on the proliferation, migration, and invasion of TPC-1 cells. Expression levels of mitogen-activated protein kinase 4 (MAPK4) and phosphorylation levels of protein kinase B (AKT), extracellular regulated protein kinase (ERK), and P38 mitogen-activated protein kinase (P38) were detected by western blotting or immunohistochemistry (IHC). Results: The expression level of miR-576-5p in PTC tissues and TPC-1 cells was significantly increased. In vitro, overexpression of miR-576-5p promoted the proliferation, migration, and invasion of TPC-1 cells. In addition, MAPK4 was highly expressed in PTC tissues, and miR-576-5p could upregulate the expression of MAPK4. Interestingly, MAPK4 knockdown reversed cell proliferation but not migration and invasion in TPC-1 cells after miR-576-5p was overexpressed. Moreover, overexpression of miR-576-5p induced activation of the AKT pathway in TPC-1 cells, and MAPK4 gene knockout reversed this AKT pathway activation. Conclusion: In this study, we found that miR-576-5p was significantly overexpressed in PTC tissues and TPC-1 cells. In addition, miR-576-5p promoted the proliferation of TPC-1 cells by enhancing expression of MAPK4 and activating the AKT pathway.

Prediction of Metastasis in the Axillary Lymph Nodes of Patients With Breast Cancer: A Radiomics Method Based on Contrast-Enhanced Computed Tomography.

BACKGROUND: The use of traditional techniques to evaluate breast cancer is restricted by the subjective nature of assessment, variation across radiologists, and limited data. Radiomics may predict axillary lymph node metastasis (ALNM) of breast cancer more accurately. PURPOSE: The aim was to evaluate the diagnostic performance of a radiomics model based on ALNs themselves that used contrast-enhanced computed tomography (CECT) to detect ALNM of breast cancer. METHODS: We retrospectively enrolled 402 patients with breast cancer confirmed by pathology from January 2016 to October 2019. Three hundred and ninety-six features were extracted for all patients from axial CECT images of 825 ALNs using Artificial Intelligent Kit software (GE Medical Systems, Version V3.1.0.R). Next, the radiomics model was trained, validated, and tested for predicting ALNM in breast cancer by using a support vector machine algorithm. Finally, the performance of the radiomics model was evaluated in terms of its classification accuracy and the value of the area under the curve (AUC). RESULTS: The radiomics model yielded the best classification accuracy of 89.1% and the highest AUC of 0.92 (95% CI: 0.91-0.93, p=0.002) for discriminating ALNM in breast cancer in the validation cohorts. In the testing cohorts, the model also demonstrated better performance, with an accuracy of 88.5% and an AUC of 0.94 (95% CI: 0.93-0.95, p=0.005) for predicting ALNM in breast cancer. CONCLUSION: The radiomics model based on CECT images can be used to predict ALNM in breast cancer and has significant potential in clinical noninvasive diagnosis and in the prediction of breast cancer metastasis.

Identifying Periampullary Regions in MRI Images Using Deep Learning.

BACKGROUND: Development and validation of a deep learning method to automatically segment the peri-ampullary (PA) region in magnetic resonance imaging (MRI) images. METHODS: A group of patients with or without periampullary carcinoma (PAC) was included. The PA regions were manually annotated in MRI images by experts. Patients were randomly divided into one training set, one validation set, and one test set. Deep learning methods were developed to automatically segment the PA region in MRI images. The segmentation performance of the methods was compared in the validation set. The model with the highest intersection over union (IoU) was evaluated in the test set. RESULTS: The deep learning algorithm achieved optimal accuracies in the segmentation of the PA regions in both T1 and T2 MRI images. The value of the IoU was 0.68, 0.68, and 0.64 for T1, T2, and combination of T1 and T2 images, respectively. CONCLUSIONS: Deep learning algorithm is promising with accuracies of concordance with manual human assessment in segmentation of the PA region in MRI images. This automated non-invasive method helps clinicians to identify and locate the PA region using preoperative MRI scanning.

Histone demethylase KDM5A promotes tumorigenesis of osteosarcoma tumor.

Osteosarcoma is a primary bone malignancy with a high rate of recurrence and poorer prognosis. Therefore, it is of vital importance to explore novel prognostic molecular biomarkers and targets for more effective therapeutic approaches. Previous studies showed that histone demethylase KDM5A can increase the proliferation and metastasis of several cancers. However, the function of KDM5A in the carcinogenesis of osteosarcoma is not clear. In the current study, KDM5A was highly expressed in osteosarcoma than adjacent normal tissue. Knockdown of KDM5A suppressed osteosarcoma cell proliferation and induced apoptosis. Moreover, knockdown of KDM5A could increase the expression level of P27 (cell-cycle inhibitor) and decrease the expression of Cyclin D1. Furthermore, after knockout of KDM5A in osteosarcoma cells by CRISPR/Cas9 system, the tumor size and growth speed were inhibited in tumor-bearing nude mice. RNA-Seq of KDM5A-KO cells indicated that interferon, epithelial-mesenchymal transition (EMT), IL6/JAK/STAT3, and TNF-α/NF-κB pathway were likely involved in the regulation of osteosarcoma cell viability. Taken together, our research established a role of KDM5A in osteosarcoma tumorigenesis and progression.

Transfer of heavy metal along food chain: a mini-review on insect susceptibility to entomopathogenic microorganisms under heavy metal stress.

The development and physiological status of pest insects are important factors that affect the effectiveness of biological control. Current knowledge reveals that heavy metals can be transferred to phytophagous insects through food chains and cause various chronic toxicological effects on the growth and physiology of phytophagous insects. These findings potentially attribute heavy metal contamination to an environmental factor governing biocontrol efficiency against pest insects, pointing to an urgent demand to better understand the effects of heavy metal exposure on insect susceptibility to entomopathogenic microorganisms. Here we discuss the transfer characteristics of heavy metals along the food chains to phytophagous insects and conclude that heavy metal exposure may promote insect susceptibility to entomopathogenic microorganisms in the heavy metal-contaminated regions. Furthermore, we propose a 'combined effect' hypothesis that combination of entomopathogenic agent and heavy metal stress can cause a much higher overall insect mortality than does the entomopathogenic agent or the heavy metal stress alone. This is a new and relatively unexplored area in the microbial-based biocontrol research, which might have great potential for future optimization of biocontrol strategies against economically and ecologically important agricultural or forest pests in the heavy metal polluted areas. © 2020 Society of Chemical Industry.

Blocking retrograde axonal transport of autophagosomes contributes to sevoflurane-induced neuron apoptosis in APP/PS1 mice.

Autophagy, a crucial pathway for the degradation of proteins in eukaryotic cells, is linked to the development of Alzheimer's disease (AD), and the accumulated autophagosomes in the cells resulting in the death of cells. Sevoflurane can impair spatial learning and memory in mice with AD and lead to the apoptosis of nerve cells; however, the underlying mechanisms remain unknown. We aim to explore the effects and underlying mechanisms of sevoflurane in APPswe/PS1ΔE9 double-transgenic mice. 51 heterozygous APPswe/PS1ΔE9 double-transgenic mice were involved and divided into three groups, including control group, sham group and sevoflurane group. Morris water maze experiment was used to test the learning and memory abilities of mice, flow cytometry was conducted to detect apoptosis and mitochondrial membrane potential of brain cells in mice, transmission electron microscopy was used to observe the number of autophagosomes at the axon in mice, and western blot was carried out to detect the expression of Bax, Bcl-2, LC3II, P62, KIF3B and DIC proteins of brain cells in mice. In our study, we found that significantly longer escape latencies, fewer crossings of the platform and shorter time spent in the target quadrant of the morris water maze experiment in the sevoflurane group. Flow cytometry showed cellular apoptosis was increased and the membrane potential of the mitochondria was reduced of brain cells in the sevoflurane group. Transmission electron microscopy displayed that there was a remarkable upregulation of autophagosomes at the axon of brain cells in mice after treatment of sevoflurane. Western blot demonstrated that the expression of Bax, LC3II, P62 and KIF3B proteins were elevated, and the expression of Bcl-2 and DIC proteins were reduced in the sevoflurane group. Sevoflurane impaired acquisition learning and memory function, promoted the apoptosis of hippocampal neurons in APPswe/PS1ΔE9 double-transgenic mice, and the mechanism might be related to the activation of autophagy along with the disruption of autophagosomes retrograde transport in axons.

Astragaloside IV enhanced carboplatin sensitivity in prostate cancer by suppressing AKT/NF-κB signaling pathway.

In our study, we explored the effect of astragaloside IV (AgIV) on carboplatin chemotherapy in prostate cancer cell lines in vitro and in vivo. Cell viability assay, colony formation assay, flow cytometry, Western blot, immunohistochemistry, immunofluorescence, and tumor xenograft growth assay were conducted. We found that AgIV significantly decreased the half-maximal inhibitory concentration of carboplatin in prostate cancer cell lines LNCap and PC-3. Moreover, AgIV enhanced the effect of carboplatin in suppressing colony formation and inducing cell apoptosis. A low-dose carboplatin treatment upregulated N-cadherin and Vimentin expression and downregulated E-cadherin expression, but this effect was abolished by combining with AgIV. Carboplatin treatment increased the levels of p-AKT and p-p65 and decreased p-IκBα, but AgIV treatment suppressed this. In addition, AgIV synergized with carboplatin to suppress tumor xenograft growth of PC-3 cells, and decreased pAKT and p-p65 levels in vivo. Our results suggested that AgIV enhanced carboplatin sensitivity in prostate cancer cell lines by suppressing AKT/NF-κB signaling, thus suppressed epithelial-mesenchymal transition induced by carboplatin. Our findings provided a new mechanism for AgIV in overcoming drug resistance of platinum-based chemotherapy and suggested a potential combination therapy of AgIV and carboplatin in prostate cancer.

Mycorrhizal type influences plant density dependence and species richness across 15 temperate forests.

Recent studies suggest that the mycorrhizal type associated with tree species is an important trait influencing ecological processes such as response to environmental conditions and conspecific negative density dependence (CNDD). However, we lack a general understanding of how tree mycorrhizal type influences CNDD strength and the resulting patterns of species abundance and richness at larger spatial scales. We assessed 305 species across 15 large, stem-mapped, temperate forest dynamics plots in Northeastern China and North America to explore the relationships between tree mycorrhizal type and CNDD, species abundance, and species richness at a regional scale. Tree species associated with arbuscular mycorrhizal (AM) fungi showed a stronger CNDD and a more positive relationship with species abundance than did tree species associated with ectomycorrhizal (ECM) fungi. For each plot, both basal area and stem abundance of AM tree species was lower than that of ECM tree species, suggesting that AM tree species were rarer than ECM tree species. Finally, ECM tree dominance showed a negative effect on plant richness across plots. These results provide evidence that tree mycorrhizal type plays an important role in influencing CNDD and species richness, highlighting this trait as an important factor in structuring plant communities in temperate forests.

Cd exposure-induced growth retardation involves in energy metabolism disorder of midgut tissues in the gypsy moth larvae.

Cadmium, a common environmental contaminant in both terrestrial and aquatic ecosystems, presented a serious hazard to growth and development of phytophagous insects. For better understanding the toxicology of Cd exposure on phytophagous insects, the physiological and molecular mechanisms underlying the energy metabolism disorder in midgut tissue of gypsy moth larvae fed on Cd-amended artificial diets (3.248 or 44.473 mg Cd/kg fresh food) were investigated. Our results showed that compared with control, Cd exposure at both two levels triggered detriment effects on growth indexes, and with the increase of exposure concentrations, the adverse effects were significantly exacerbated. Larval growth and nutritional indexes (except approximate digestibility) showed a strong positive correlation, indicating that growth retardation in the gypsy moth larvae under Cd stress was tightly related to the food utilization. The key genes at mRNA level in glycolysis/gluconeogenesis, citrate cycle pathway and starch/sucrose metabolism pathway also presented a significant and positive correlation with growth indexes, once again demonstrating that energy metabolism was the key factor that controls the growth and development of the gypsy moth larvae under Cd stress. Antioxidant system collapse and oxidative damage, a chief cause of histopathological alterations in midgut tissue, consist of the physiological basis of energy metabolism disorder in Cd-treated gypsy moth larvae. Together, these results suggest that histopathological alterations or oxidative damage of tissue structure significant disturbed physiological functions of midgut tissue in gypsy moth larvae exposed to Cd stress, as reflected via food utilization or energy metabolism disorder, and eventually resulted in larval growth retardation.

Solid pseudopapillary neoplasm (SPN) of the pancreas presenting with ascites misdiagnosed as pancreatic tuberculosis: a case report and literature review.

INTRODUCTION: Solid pseudopapillary neoplasm (SPN) is a rare pancreatic tumor that mainly affects young women. It is a low-grade malignant neoplasm, with an excellent prognosis after surgical treatment. We report herein a case of SPN presenting with ascites that was misdiagnosed as pancreatic tuberculosis (TB). CASE REPORT: A 16-year-old female initially presented with a large volume of ascites. Contrast-enhanced ultrasound and computed tomography found a heterogeneous lesion in the pancreatic body, which had slight contrast enhancement on the arterial phase. Analysis of ascites showed it was exudative. Endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) of the mass only revealed a few blood clots. The diagnosis was highly suggestive of a pancreatic TB. However, after 6 months of anti-TB therapy, the pancreatic lesion remained essentially unchanged. Subsequently, magnetic resonance imaging indicated a mixed solid and cystic lesion with a well-defined margin in the pancreatic body. Further EUS-FNA showed monomorphic neoplastic cells with papillary architecture and immunohistochemical analysis revealed that the tumor cells were positive for ß-catenin, CD10, vimentin, cytokeratin, and synaptophysin. These findings were consistent with SPN. After distal pancreatectomy with splenectomy, postoperative pathology and immunohistochemical staining confirmed the diagnosis of SPN. CONCLUSION: Clinicians should consider the possibility of SPN for pancreatic heterogeneous masses. Multiple diagnostic imaging modalities and EUS-FNA may contribute to the preoperative diagnosis of this disease.

The effect of hepatocellular carcinoma-associated fibroblasts on hepatoma vasculogenic mimicry.

Vasculogenic Mimicry (VM) is the main source of blood supply in the early stage of tumor growth. Carcinoma-associated fibroblasts (CAFs) are one of the most important host cells in the tumor microenvironment. Some studies have found that CAFs can promote tumor angiogenesis, but there are few reports on the relationship between CAFs and VM. Tissue samples were collected from 60 cases of hepatocellular carcinoma (HCC) and 10 persons with normal liver function. The relationship between VM expression and clinicopathologic features was analyzed. Furthermore, the relationship between VM expression and vimentin or α-SMA expression was analyzed. Primary culture of hepatocellular CAFs and the collection of conditioned media were carried out. The effects of hepatocellular CAF conditioned medium on the formation of VM and the levels of VM-related proteins and genes in MHCC-97H cells were studied. The positive rate of VM was 35.0% in HCC tissues. There was no VM expression in normal liver tissues. VM expression was related to tumor diameter, Edmondson grade, clinical stage, and liver cirrhosis. The expression of vimentin and α-SMA in VM-positive patients was higher than in VM-negative patients. Different concentrations of hepatocellular CAF conditioned medium could promote the formation of VM and increase the expression of VM-related genes and proteins (MMP2 and EphA2) in MHCC-97H cells. The results show that there was a significant correlation between VM formation and the expression of vimentin or α-SMA in HCC tissues. The conditioned medium of hepatocellular CAFs may promote VM formation and the expression of VM-related genes and proteins (MMP2 and EphA2) in hepatoma cell line MHCC-97H.

Differential responses to Cd stress induced by exogenous application of Cu, Zn or Ca in the medicinal plant Catharanthus roseus.

Cd(II) is one of the most widespread and toxic heavy metals and seriously threatens plant growth, furthermore negatively affecting human health. For survival from this metal stress, plants always fight with Cd(II) toxicity by themselves or using other external factors. The effects of second metals copper (Cu(II)), zinc (Zn(II)) and calcium (Ca(II)) on the Cd(II)-affected root morphology, Cd(II) translocation and metabolic responses in Catharanthus roseus were investigated under hydroponic conditions. We found that the Cd-stressed plants displayed the browning and rot root symptom, excess H2O2 content, lipid peroxidation and Cd(II) accumulation in plants. However, the supplement with second metals largely alleviated Cd-induced toxicity, including browning and rot roots, oxidative stress and internal Cd(II) accumulation. The amended effects at metabolic and transcriptional levels involved in different second metals share either common or divergent strategies. They commonly repressed Cd uptake and promoted Cd(II) translocation from roots to shoots with divergent mechanisms. High Zn(II) could activate MTs expression in roots, while Cu(II) or Ca(II) did not under Cd(II) stress condition. The presence of Ca(II) under Cd stress condition largely initiated occurrence of lateral roots. We then grouped a metabolic diagram integrating terpenoid indole alkaloid (TIA) accumulation and TIA pathway gene expression to elucidate the metabolic response of C. roseus to Cd(II) alone or combined with second metals. The treatment with 100 Cd(II) alone largely promoted accumulation of vinblastine, vindoline, catharanthine and loganin, whereas depressed or little changed the expression levels of genes detected here, compared to 0 Cd(II) control. In the presence of Cd(II), the supplement with second metals displayed specific effect on different alkaloid. Among them, the metal Ca(II) is especially beneficial for serpentine accumulation, Zn(II) mainly promoted tabersonine production. However, the addition of Cu(II) commonly depressed accumulation of most alkaloids detected here. Generally, we presented different mechanisms by which the second metals used to alleviate Cd (II) toxicity. This plant has potential application in phytoremediation of Cd(II), due to relatively substantial accumulation of biomass, as well as secondary metabolites TIAs used as pharmaceutical materials when facing Cd stress.

Metabolomics Characterization of Two Apocynaceae Plants, Catharanthus roseus and Vinca minor, Using GC-MS and LC-MS Methods in Combination.

Catharanthus roseus (C. roseus) and Vinca minor (V. minor) are two common important medical plants belonging to the family Apocynaceae. In this study, we used non-targeted GC-MS and targeted LC-MS metabolomics to dissect the metabolic profile of two plants with comparable phenotypic and metabolic differences. A total of 58 significantly different metabolites were present in different quantities according to PCA and PLS-DA score plots of the GC-MS analysis. The 58 identified compounds comprised 16 sugars, eight amino acids, nine alcohols and 18 organic acids. We subjected these metabolites into KEGG pathway enrichment analysis and highlighted 27 metabolic pathways, concentrated on the TCA cycle, glycometabolism, oligosaccharides, and polyol and lipid transporter (RFOS). Among the primary metabolites, trehalose, raffinose, digalacturonic acid and gallic acid were revealed to be the most significant marker compounds between the two plants, presumably contributing to species-specific phenotypic and metabolic discrepancy. The profiling of nine typical alkaloids in both plants using LC-MS method highlighted higher levels of crucial terpenoid indole alkaloid (TIA) intermediates of loganin, serpentine, and tabersonine in V. minor than in C. roseus. The possible underlying process of the metabolic flux from primary metabolism pathways to TIA synthesis was discussed and proposed. Generally speaking, this work provides a full-scale comparison of primary and secondary metabolites between two medical plants and a metabolic explanation of their TIA accumulation and phenotype differences.

Effects of TGF-ß1 on the Proliferation and Apoptosis of Human Cervical Cancer Hela Cells In Vitro.

To investigate the effects of TGF-ß1 on the proliferation and apoptosis of cervical cancer Hela cells in vitro. Human cervical cancer Hela cells were cultured in vitro and divided into the experimental and control groups. In the experimental groups, Hela cells were stimulated with different concentrations of TGF-ß1 (0.01, 0.1, 1, and 10 ng/mL), while Hela cells cultured in serum-free medium without TGF-ß1 were used as controls. The CCK8 method was adopted to detect the effect of TGF-ß1 on Hela cell proliferation, and flow cytometry was used to determine cell apoptosis 72 h after TGF-ß1 treatment. Compared with the control group, the CCK-8 tests showed that different concentrations of TGF-ß1 had no obvious effect on Hela cell proliferation 24 h after treatment (P > 0.05). However, upon 48 or 72 h of treatment, TGF-ß1 significantly inhibited the proliferation of Hela cells in a time- and dose-dependent manner (P < 0.05). The flow cytometry results indicated that TGF-ß1 influenced the apoptosis of human cervical cancer Hela cells in a dose-dependent manner after 72 h of treatment (P < 0.05). TGF-ß1 significantly inhibited the growth and induced the apoptosis of human cervical Hela cells in vitro.