[Influence of climate change and human activities on grassland phenology in Anhui Province]. / 安徽省气候变化和人类活动对草地物候的影响.

To explore the influence of climate change and human activities on grassland phenology in Anhui Pro-vince, and quantify the contribution rate of climate change and human activities to phenology, we extracted the phenology of grassland, including the start of growing season (SOS) and the end of growing season (EOS), based on the normalized difference vegetation index (NDVI) dataset of Anhui Province from 2003 to 2020. The temporal and spatial characteristics and future evolution trends of phenological changes were analyzed using slope trend ana-lysis, Mann-Kendall non-parametric test, and Hurst index. We further conducted correlation analysis and residual analysis based on the datasets of mean annual temperature and mean annual precipitation to explore the responses of phenology to climate change and human activities, and quantify their contribution rate. The results showed that SOS and EOS showed an advancing trend with a rate of 0.8 and 0.7 days per year from 2003 to 2020. SOS in the sou-thern part of the study area was significantly earlier than in the central and northern regions, while EOS gradually advanced from south to north. Both SOS and EOS in the future showed an advancing trend. SOS was negatively correlated with annual average temperature, while positively correlated with annual precipitation. EOS was negatively correlated with both annual average temperature and annual precipitation. The proportion of the area where SOS was advanced driven by both climate change and human activities was 56.9%, and the value was 48.3% for EOS. Human activities were the main driving factor for phenology, and climate change was the secondary driving factor. The relative contributions of human activities and climate change to SOS were 66.4% and 33.6%, and to EOS were 61.2% and 38.8%, respectively. Human activities had stronger impact on SOS and EOS than climate change, resulting in earlier phenology.

Successful treatment of severe pityriasis rubra pilaris with secukinumab in a 3-year-old boy.

Pityriasis rubra pilaris (PRP) is a rare, scaly, keratotic inflammatory skin disease characterized by red scaly patches, keratosis papules, palmoplantar keratoderma and scaling of the scalp. In severe cases, ectropion of the eyelid may occur, and erythroderma may further develop. Recently, it has been reported that secukinumab, a monoclonal anti-interleukin-17A antibody, has certain efficacy in the treatment of PRP. Herein, we report a 3-year-old Chinese boy with severe Type III (classic juvenile) PRP who was successfully treated with secukinumab alone.

Exosomal miR-3682-3p Suppresses Angiogenesis by Targeting ANGPT1 via the RAS-MEK1/2-ERK1/2 Pathway in Hepatocellular Carcinoma.

BACKGROUND: Angiogenesis is a crucial process in tumorigenesis and development. The role of exosomes derived from hepatocellular carcinoma (HCC) cells in angiogenesis has not been clearly elucidated. METHODS AND RESULTS: Exosomes were isolated from HCC cell lines (HCCLM3, MHCC97L, and PLC/RFP/5) by ultracentrifugation and identified by nano transmission electron microscopy (TEM), NanoSight analysis and western blotting, respectively. In vitro and in vivo analyses showed that exosomes isolated from highly metastatic HCC cells enhanced the migration, invasion and tube formation of human umbilical vein endothelial cells (HUVECs) compared to exosomes derived from poorly metastatic HCC cells. In addition, microarray analysis of HCC-Exos was conducted to identify potential functional molecules, and miR-3682-3p expression was found to be significantly downregulated in exosomes isolated from highly metastatic HCC cells. By in vitro gain-of-function experiments, we found that HCC cells secreted exosomal miR-3682-3p, which negatively regulates angiopoietin-1 (ANGPT1), and this led to inhibition of RAS-MEK1/2-ERK1/2 signaling in endothelial cells and eventually impaired angiogenesis. CONCLUSION: Our study elucidates that exosomal miR-3682-3p attenuates angiogenesis by targeting ANGPT1 through RAS-MEK1/2-ERK1/2 signaling and provides novel potential targets for liver cancer therapy.

[The Application of Estrogen Receptor Aptamer in Immunohistochemical Detection of Breast Cancer].

OBJECTIVE: To synthesize and select an estrogen receptors aptamer that can be used in immunostaining of breast cancer tissues. METHODS: ER protein was purified. ER aptamer that showed a high affinity and specificity for ER was synthesized and selected and by SELEX. Ligand -receptor interactions assay was adopted to measure the affinity of the aptamer-ER complex. Both the biotinylated aptamer and the anti-ER monoclonal antibody were tested for immunohistochemical staining of ER status on 105 breast cancer samples. Agreement on the detection of ER expression was determined by Kappa statistics. RESULTS: The dissociation contant (Kd) of the biotinylated aptamer-ER complex, as calculated by a linear regression analysis, was determined to be (0.34±0.05) nmol/L ( n=3, r=0.989). The binding capacity (B max) was 769.23 fmol/(mg prot·nmol -1·L -1). The ER aptamer and the anti-ER antibody both exhibited identical specificity to ER-expressing breast cancer cells. There was a high agreement between the two methods ( n=105, Kappa value=0.943, 95% confident interval=0.879-1.006, P<0.05 for the ER positive and negtive samples; n=75, Kappa value=0.805, 95% confident interval=0.642-0.967, P<0.05 for the ER weak and moderate/strong expression samples). Both the anti-ER antibody and the ER aptamer can also recognized breast cancer cells at the same sites. There was no expression in the negative controls. There were also positive expressions in the 2 endometrial cancer tissues by using biotinylated aptamer. CONCLUSIONS: Our results indicated that the synthesized ER aptamer has a high affinity to bind ER. ER aptamer and the anti-ER antibody can both be used for immunohistochemical staining of ER status in breast cancer tissue.

Development of a novel anti-human aspartyl-(asparaginyl) ß-hydroxylase monoclonal antibody with diagnostic and therapeutic potential.

Human aspartyl-(asparaginyl)-ß-hydroxylase (HAAH) has recently been the subject of several studies, as it was previously observed to be overexpressed in numerous types of carcinoma cells and tissues in patient tumor samples. HAAH has been implicated in tumor invasion and metastasis, indicating that it may be an important target and biomarker for tumor diagnosis and treatment. However, the immunological tools currently available for the study of this protein, including monoclonal antibodies, are limited, as is the present knowledge regarding the role of HAAH in tumor therapy and diagnosis. In the present study, a recombinant C-terminal domain of HAAH was expressed in Pichia pastoris and a novel monoclonal antibody (mAb) targeting HAAH (HAAH-C) was constructed. Immunofluorescence and antibody-dependent cellular cytotoxicity (ADCC) assays were used to demonstrate the specificity and ADCC activity of this antibody. The results demonstrated that this anti-C-terminal HAAH mAB, in combination with an existing anti-N terminal HAAH mAb, exhibited a high response to native HAAH from carcinoma cell culture supernatant, as measured with a double antibody sandwich enzyme-linked immunosorbent assay. This validated novel mAB-HAAH-C may prompt further studies into the underlying mechanisms of HAAH, and the exploration of its potential in tumor diagnosis and therapy.

The anti-inflammatory effect of Sonchus oleraceus aqueous extract on lipopolysaccharide stimulated RAW 264.7 cells and mice.

CONTEXT: Sonchus oleraceus L. (Asteraceae) (SO) is a dietary and traditional medicinal plant in China. However, its underlying mechanism of action as an anti-inflammatory agent is not known. OBJECTIVE: This study evaluates the anti-inflammatory activity of aqueous extract of SO. MATERIALS AND METHODS: The extract of SO was used to treat RAW 264.7 cells (in the working concentrations of 500, 250, 125, 62.5, 31.3 and 15.6 µg/mL) for 24 h. Pro-inflammatory cytokines and mediators produced in LPS-stimulated RAW 264.7 cells were assessed. Meanwhile, the expression level of TLR-4, COX-2, pSTATs and NF-κB was tested. Moreover, the anti-inflammatory activity of the extract in vivo was assessed using xylene-induced mouse ear oedema model and the anti-inflammatory compounds in the extracts were analyzed by HPLC-MS. RESULTS: SO extract significantly inhibited the production of pro-inflammatory cytokines and mediators at gene and protein levels with the concentration of 31.3 µg/mL, and suppressed the expression of TLR-4, COX-2, NF-κB and pSTAT in RAW 264.7 cells. The anti-inflammatory activity of SO in vivo has significant anti-inflammatory effects with the concentration of 250 and 125 mg/kg, and less side effect on the weights of the mice at the concentration of 250 mg/kg. Moreover, HPLC-MS analysis revealed that the anti-inflammatory compounds in the extract were identified as villosol, ferulaic acid, ß-sitosterol, ursolic acid and rutin. DISCUSSION AND CONCLUSION: This study indicated that SO extract has anti-inflammatory effects in vitro and in vivo, which will be further developed as novel pharmacological strategies in order to defeat inflammatory diseases.

Spindle cell metaplastic carcinoma of breast: A clinicopathological and immunohistochemical analysis.

AIM: To better characterize spindle cell metaplastic carcinoma (SpCMC) of breast, a rare variant of breast cancer that has been classified under the broad rubric of metaplastic carcinoma. METHODS: We presented herein 19 cases of metaplastic breast carcinoma with dominant spindle cell component. All cases were clinically of breast origin, showed more than 80% spindle morphology, 10 cases exhibited pure spindled morphology, 8 contained invasive ductal carcinoma (IDC) and 1 presented with ductal carcinoma in situ elements. RESULTS: Immunohistochemical studies showed evidence suggesting myoepithelial and epithelial differentiation as exhibited by immunoreactivity for at least one myoepithelial and epithelial markers in all pure spindle cell components. IDC group showed 21.7% of axillary lymph nodes metastasis rate, whereas the axillary lymph node metastasis rate of the SpCMC group was 1.3%, significantly lower than that of the IDC group (P < 0.001). Immunohistochemical staining of IDC exhibited higher degrees of positivity for ER, PR and Her2 (90, 60 and 30%, respectively) when compared with the SpCMC group, which showed a positive degree of 5.2, 5.2 and 10.5% for ER, PR and Her2, respectively (P < 0.001). CONCLUSION: Based on this series, SpCMC is a rare variant of metaplastic breast carcinoma with the distinct histopathological and immunohistochemical features. The biological behaviors of SpCMC, like axillary lymph node status, were quite different from that of IDC, suggesting that it may act as an independent pathologic subtype. Immunohistochemical analysis of a panel of epithelial and myoepithelial markers could contribute to the pathologic diagnosis of SpCMC.

GPR78 promotes lung cancer cell migration and metastasis by activation of Gαq-Rho GTPase pathway.

GPR78 is an orphan G-protein coupled receptor (GPCR) that is predominantly expressed in human brain tissues. Currently, the function of GPR78 is unknown. This study revealed that GPR78 was expressed in lung cancer cells and functioned as a novel regulator of lung cancer cell migration and metastasis. We found that knockdown of GPR78 in lung cancer cells suppressed cell migration. Moreover, GPR78 modulated the formation of actin stress fibers in A549 cells, in a RhoA- and Rac1-dependent manner. At the molecular level, GPR78 regulated cell motility through the activation of Gαq-RhoA/Rac1 pathway. We further demonstrated that in vivo, the knockdown of GPR78 inhibited lung cancer cell metastasis. These findings suggest that GPR78 is a novel regulator for lung cancer metastasis and may serve as a potential drug target against metastatic human lung cancer. [BMB Reports 2016; 49(11): 623-628].

Heat shock protein 90 inhibitors induce functional inhibition of human natural killer cells in a dose-dependent manner.

Heat shock protein 90 (Hsp90) is a ubiquitously expressed ATP-dependent molecular chaperone across all species that helps to the correct the folding of many proteins related to important signaling pathways. Tumor cells expressing Hsp90 have more ATP-binding affinity than normal cells. Many correlative inhibitors have been developed to promising anti-tumor strategies and have been evaluated in clinical trials. However, the effect of Hsp90 inhibitors on immunocytes cannot be ignored. Natural killer (NK) cells are key components of the innate immune system that play a pivotal role in tumor surveillance. The present study has investigated the potential effect of four Hsp90 inhibitors (NVP-AUY922, BIIB021, 17-DMAG, and SNX-2112) on human primary NK cells. The viability, cytotoxicity, apoptosis, phenotype, and cytokine secretion of NK cells after inhibitor treatment were assessed. The results of this study demonstrated that the inhibitors had negative effects on NK cell activity in a dose-dependent manner. The four inhibitors significantly reduced the cytotoxicity of the NK cells by decreasing viability, inducing apoptosis and down-regulating the expression of cytokines and functional receptors. These findings suggest that more attention should be given to the effect of Hsp90 inhibitors on NK cell function during clinical trials and also represent a potential immunosuppressant strategy.

ADAM15 targets MMP9 activity to promote lung cancer cell invasion.

ADAM15 is a membrane-associated proteinase belonging to a disintegrin and metalloproteinase (ADAM) family. Recent studies suggested that ADAM15 is overexpressed in several types of cancer and is involved in metastatic tumor progression. However, the function of ADAM15 in non-small cell lung cancer (NSCLC) is currently unknown. In the present study, we found that high expression of ADAM15 was associated with decreased overall survival (OS) and disease-free survival (DFS) in NSCLC patients. Furthermore, shRNA-mediated knockdown of ADAM15 attenuated cell migration and invasion. Mechanistic study demonstrated that ADAM15 upregulated MMP9 expression in lung cancer cells via activation of the MEK-ERK pathway. Moreover, ADAM15 proteolytically cleaved and activated pro-MMP9 in vitro and interacted with MMP9 in vivo. Overexpression of ADAM15 in A549 cells promoted cell invasion, while knocking down MMP9 attenuated cell invasive ability. Therefore, our data not only support a pro-metastatic role of ADAM15 in lung cancer progression, but also reveal a novel mechanism of ADAM15 in promoting cancer cell invasion through directly targeting MMP9 activation.

Using a sequence of estrogen response elements as a DNA aptamer for estrogen receptors in vitro.

Estrogen receptors (ERs) are overexpressed in approximately 70% of breast cancer cases, and they play an important role in tumorigenesis. ERs are strong predictive factors for measuring responses to hormonal therapies. Aptamers are short and single stranded oligonucleotides that are able to recognize target molecules with high affinity. In the present study, we selected and synthesized an oligonucleotide, which has a similar sequence to estrogen response element in the Xenopus Vitellogenin A2 gene. The synthesized oligonucleotide was evaluated by using immunostaining of paraffin-embedded breast cancer tissues and treating MCF-7 human mammary carcinoma cell line in vitro. We found that the synthesized oligonucleotide had a high binding affinity to ER similar to estradiol. Using a specific anti-ER antibody as a standard control, we showed that the synthesized oligonucleotide specifically recognized and immunostained tumor cells of breast cancer without cross-reaction with normal tissues. The overall agreement of ER detection between the anti-ER antibody and the ER aptamer was 97.1% (kappa value=0.943; 95% CI=0.879-1.006; p<0.002). Similar to tamoxifen or fulvestrant, the oligonucleotide also had an inhibitory effect on cell proliferation of MCF-7 cell line in a dose- and time-dependent fashion but had no cytotoxic effect on human normal mammary epithelial cells. Therefore, the synthesized oligonucleotide may be used as an aptamer for immunostaining of paraffin-embedded tissue sections for breast cancer diagnosis, as well as a potential ER antagonist in the treatment of breast cancer.

Speckle-tracking imaging to monitor myocardial function after coronary artery bypass graft surgery.

OBJECTIVES: The purpose of this study was to investigate the changes in myocardial function in patients after coronary artery bypass graft (CABG) surgery using longitudinal and circumferential strain on speckle-tracking imaging. METHODS: A total of 145 patients who successfully underwent CABG surgery with a left ventricular ejection fraction (LVEF) of 50% or greater were enrolled in this study. Patients were classified into 4 groups based on age: group 1 (33-59 years), group 2 (60-64 years), group 3 (65-69 years), and group 4 (70-79 years). Routine echocardiography and longitudinal and circumferential strain measurements on speckle-tracking imaging were performed 1 week before and 1, 3, and 6 months after the CABG. RESULTS: In all groups, longitudinal strain increased significantly at 3 and 6 months after CABG therapy compared to baseline (P < .05). A significant increase in circumferential strain was found 1 month after the CABG in groups 1, 2, and 3, and a continuous increase in the parameter was observed in all groups 3 months after therapy (P < .05). However, the LVEF, left ventricular end-diastolic dimension, and stroke volume measured by routine echocardiography were not significantly changed after successful CABG treatment in all groups during 6 months of follow-up. CONCLUSIONS: Based on the results of our study in all age groups, speckle-tracking imaging parameters are more effective than the LVEF, left ventricular end-diastolic dimension, and stroke volume for monitoring improvement in myocardial function after CABG surgery.

Relationship between epicardial adipose tissue volume measured using coronary computed tomography angiography and atherosclerotic plaque characteristics in patients with severe coronary artery stenosis.

AIM: To investigate the relationship between epicardial adipose tissue (EAT) volume and coronary plaque composition. METHODS: EAT volume (measured using coronary computed tomography angiography) and coronary plaque characteristics were assessed on a per-segment basis (modified 15-segment American College of Cardiology/American Heart Association classification) in patients with severe coronary artery stenosis. Coronary plaques were classified into four types: 1, calcified plaques; 2, mixed plaques (calcification dominant); 3, mixed plaques (non-calcification dominant); and 4, non-calcified plaques. The gold standard for luminal stenosis was conventional coronary angiography. RESULTS: In 365 patients (mean age 58.7 ± 8.0 years), EAT volume was 169.85 ± 29.94 ml. There were no significant between-group differences in patient characteristics. Statistically significant differences in EAT volume between non-calcified and calcified plaque groups were observed. EAT volume showed a positive correlation with total cholesterol and low-density-lipoprotein cholesterol, and a very weak correlation with age, triglyceride, body mass index and high-density-lipoprotein cholesterol. CONCLUSION: EAT volume was higher in the presence of non-calcified and mixed plaques in patients with ≥50% severe coronary artery stenosis.

[The research progress of Aurora-B kinase and its inhibitors].

Aurora-B as an important kinase to adjust the cell normal mitosis is a potent target for cancer treatment. Aurora-B is overexpressed in a broad range of tumor and tumor cells are more sensitive while Aurora-B is inhibited. Due to the key role of the Aurora-B in cell mitosis, the development of its inhibitors is becoming more and more important. Several small molecules inhibit with a similar efficacy both Aurora-A and Aurora-B, however, in most cases the effects resemble Aurora-B disruption by genetic methods, indicating that Aurora-B represents an effective therapeutic target. There were several Aurora-B kinase inhibitors which had entered the clinics and displayed good antitumor activity. In this review, we will outline the functions of Aurora kinase B in normal cell division and in malignancy. We will focus on recent preclinical and clinical studies that have explored the mechanism of action and clinical effect of Aurora-B inhibitors in cancer treatment.

Importance of HLA-G expression and tumor infiltrating lymphocytes in molecular subtypes of breast cancer.

This study is aimed at investigating whether or not human leukocyte antigen-G (HLA-G) expression is associated with breast cancer molecular subtypes. HLA-G expression was immunohistochemically investigated in 104 patients with invasive ductal breast carcinoma, in which 56 were luminal A, 17 were luminal B, 19 were HER-2, and 12 were basal-like/normal breast-like subtype classified according to immunohistochemical staining results of ER, HER-2, CK5/6, and EGFR. Host immune response status was assessed by estimating the density of tumor infiltrating lymphocytes (TIL). For comparison, other biomarkers such as Ki67, p53 and VEGF were also investigated. Associations of these biomarkers and TIL with molecular subtypes were statistically analyzed. We found that there were more cases with high expressions of HLA-G in non-luminal than in luminal subtypes (P=0.035). In contrast, more cases with high density of TIL was found in luminal than in non-luminal subtypes (P=0.023). Compared to all the biomarkers studied, only HLA-G expression was found to be inversely associated with the density of TIL (P=0.004). Furthermore, patients with HLA-G(high)/TIL(low) status had a higher risk of recurrence than those with HLA-G(low)/TIL(high) status, regardless of the molecular subtypes. Therefore, a combination of the status of HLA-G and TIL could improve the prognostic prediction for patients with various molecular subtypes of breast cancer.

Chemical characteristics and antioxidant properties of crude water soluble polysaccharides from four common edible mushrooms.

Four crude water soluble polysaccharides, CABP, CAAP, CFVP and CLDP, were isolated from common edible mushrooms, including Agaricus bisporus, Auricularia auricula, Flammulina velutipes and Lentinus edodes, and their chemical characteristics and antioxidant properties were determined. Fourier Transform-infrared analysis showed that the four crude polysaccharides were all composed of ß-glycoside linkages. The major monosaccharide compositions were D-galactose, D-glucose and D-mannose for CABP, CAAP and CLDP, while CFVP was found to consist of L-arabinose, D-galactose, D-glucose and D-mannose. The main molecular weight distributions of CABP and the other three polysaccharides were <5.1 × 10(4) Da and >66.0 × 10(4) Da, respectively. Antioxidant properties of the four polysaccharides were evaluated in in vitro systems and CABP showed the best antioxidant properties. The studied mushroom species could potentially be used in part of well-balanced diets and as a source of antioxidant compounds.

In situ follicular lymphoma with progressive transformation of the germinal centers confirmed by laser capture microdissection, IGH gene rearrangement analysis, and fluorescence in situ hybridization for t(14;18).

The authors report an unusual case of in situ follicular lymphoma associated with progressive transformation of the germinal centers. The patient was a 74-year-old Chinese woman with sequential lymphadenopathy in the right and left cervical regions over a period of 2 months. The first biopsy revealed in situ follicular lymphoma with progressive transformation of germinal centers, and the biopsy of the second lymph node led to a diagnosis of in situ follicular lymphoma. The immunophenotype, polymerase chain reaction amplification of the immunoglobulin heavy chain gene, and fluorescence in situ hybridization for t(14;18) were analyzed in each biopsy specimen, which showed both specimens to have t(14;18)(q32;q21) and revealed progression from polyclonality to monoclonality. These findings suggest a case of multicentric in situ follicular lymphoma and provide new insights into the pathogenesis of this disease.

[Expression of DOG-1 in gastrointestinal stromal tumor and its diagnostic application].

OBJECTIVE: To investigate the expression of DOG-1 in gastrointestinal stromal tumors (GIST) and its diagnostic application. METHODS: Immunohistochemical EnVision technique was used to assess the expression of DOG-1 in 84 cases of GIST in comparison with CD117 and CD34. RESULTS: All 84 cases of GIST consisted of variable proportions of spindle and epithelioid tumor cells or just one type of the tumor cell. The expression rates of DOG-1, CD117 and CD34 were 91.3% (42/46), 95.7% (44/46) and 82.6% (38/46), in the group of very low and low risk GIST, and were 100% (38/38), 100% (38/38) and 78.9% (30/38), respectively, in the group of moderate and high risk GIST. True leiomyomas, schwannomas, fibromatosis and normal gastrointestinal mucoca did not express these markers. Moreover, the sensitivity and specificity of DOG-1 in the detection of GIST were similar to those of CD117, without statistical difference (P > 0.05) between the two markers. However, the sensitivity and specificity of DOG-1 detection of moderate and high risk GIST were significantly higher than those of CD34 (P < 0.01). CONCLUSIONS: DOG-1 is a novel marker of gastrointestinal stromal tumors. It has the sensitivity and specificity higher than CD34, especially in the detection of moderate and high risk GIST. Combined DOG-1 and CD117 immunohistochemistry will likely improve the diagnostic accuracy of GIST.