MiR-26a enhances invasive capacity by suppressing GSK3ß in human lung cancer cells.

Lung cancer is the common cause of death from cancer, and most lung cancer patients die of metastasis. MicroRNAs (miRNAs) function as either oncogenes or tumor suppressors, playing crucial role not only in tumorigenesis, but also in tumor invasion and metastasis. There are several studies showed that miR-26a is involved in carcinogenesis, however, its role in tumor metastasis need to be elucidated. In this study, we showed that ectopic expression of miR-26a enhanced migration and invasion of lung cancer cells. Glycogen synthase kinase-3ß (GSK3ß) was identified as a direct target of miR-26a. GSK3ß expression negatively correlated with miR-26a expression in lung cancer tissues. Silencing of GSK3ß achieved similar effect as miR-26a over-expression; over-expression of GSK3ß reversed the enhanced effect of miR-26a on lung cancer cell migration and invasion. Further study indicated that miR-26a increased ß-catenin expression and nuclear translocation. C-myc and cyclin D1, the downstream genes of ß-catenin, were also up-regulated by miR-26a. Furthermore, xenograft study showed that miR-26a promoted lung cancer cell growth in vivo, and suppressed GSK3ß expression. Collectively, our results demonstrated that miR-26a enhanced metastatic potential of lung cancer cells via activation of ß-catenin pathway by targeting GSK3ß, suggesting the potential applicability of miR-26a as a target for cancer treatment.

Establishment of Multiplex Loop-Mediated Isothermal Amplification for Rapid Detection of Genitourinary Mycoplasma.

BACKGROUND: Genitourinary Mycoplasma (Ureaplasma urealyticum, Mycoplasma hominis, and Mycoplasma genitalium) is a common pathogen among women, which can cause funisitis, spontaneous abortion, and low birth weight. However, current laboratory testing methods for genitourinary mycoplasma normally need complex processes, expensive devices, and qualified staffs, and there are many limits for application. Up to now, the LAMP method is a rapidly developing field because of the significance for clinical application and commercial value. Few studies have reported the use of mLAMP to detect UU, MH, and MG. In this study, a multiplex loop-mediated isothermal amplification system was developed for rapid detection of UU, MH, and MG, concurrently. METHODS: Three sets of multiplex LAMP primers were designed to specifically target urease of UU, 16S rRNA of MH, and mgpa of MG. The ratio of primer concentration was optimized. The specificity and sensitivity of multiplex LAMP were explored. Twenty-nine clinical samples were successfully used with mLAMP. RESULTS: In this study, the primer concentration in the mLAMP system was set to 1.3 µmol/L which could maintain reaction efficiency and avoid non-specific reaction. Multiplex LAMP can test UU, MG, and MH simultaneously with high specificity. Meanwhile, the sensitivity of multiplex LAMP was found to be 100 pg for UU, 100 pg for MH and 1 ng for MG, which was much higher than that of conventional PCR. Furthermore, among the 29 clinical samples, there were two positive samples determined by mLAMP, which was consistent with the PCR and sequencing results. CONCLUSIONS: The multiplex LAMP assay can potentially facilitate simultaneous detection of three kinds of mycoplasma in a large number of samples in clinic, which could be used as a primary screening method and as a supplementary method for classical methods.

MMP Gene Polymorphisms, MMP-1 -1607 1G/2G, -519 A/G, and MMP-12 -82 A/G, and Ischemic Stroke: A Meta-Analysis.

BACKGROUND: Several polymorphisms in the human gene encoding MMP have been investigated for the association with ischemic stroke (IS) in the population, of which the MMP-1 -1607 1G/2G, -519 A/G, and MMP-12 -82 A/G polymorphisms gain more and more attention; however, the results are controversial and ambiguous. We therefore carried out the meta-analysis to yield a valid conclusion. METHODS: The literature database was comprehensively searched to identify potentially eligible reports. RESULTS: A total of 15 studies with 3237 cases and 3075 controls were included in this meta-analysis. CONCLUSIONS: There was a significant association in MMP-1 -1607 1G/2G and MMP-12 -82 A/G gene polymorphisms, but none was observed in MMP-1 -519 A/G. When a subgroup analysis by ethnicity and HWE, MMP-12 -82 A/G gene polymorphisms may be a risk factor for IS in Europe. In Africa, MMP-1 -1607 1G/2G and MMP-12 -82 A/G also showed a significant effect on IS. Further investigation on a larger sample size of different ethnic populations is needed to confirm the findings.

PVT1-derived miR-1207-5p promotes breast cancer cell growth by targeting STAT6.

Accumulating evidence indicates that ectopic expression of non-coding RNAs are responsible for breast cancer progression. Increased non-coding RNA PVT1, the host gene of microRNA-1207-5p (miR-1207-5p), has been associated with breast cancer proliferation. However, how PVT1 functions in breast cancer is still not clear. In this study, we show a PVT1-derived microRNA, miR-1207-5p, that promotes the proliferation of breast cancer cells by directly regulating STAT6. We first confirm the positive correlated expression pattern between PVT1 and miR-1207-5p by observing consistent induced expression by estrogen, and overexpression in breast cancer cell lines and breast cancer patient specimens. Moreover, silence of PVT1 also decreased miR-1207-5p expression. Furthermore, increased miR-1207-5p expression promoted, while decreased miR-1207-5p expression suppressed, cell proliferation, colony formation, and cell cycle progression in breast cancer cell lines. Mechanistically, a novel target of miR-1207-5p, STAT6, was identified by a luciferase reporter assay. Overexpression of miR-1207-5p decreased the levels of STAT6, which activated CDKN1A and CDKN1B to regulate the cell cycle. We also confirmed the reverse correlation of miR-1207-5p and STAT6 expression levels in breast cancer samples. Therefore, our findings reveal that PVT1-derived miR-1207-5p promotes the proliferation of breast cancer cells by targeting STAT6, which in turn controls CDKN1A and CDKN1B expression. These findings suggest miR-1207-5p might be a potential target for breast cancer therapy.

Effect of Water Chemistry and Hydrodynamics on Nitrogen Transformation Activity and Microbial Community Functional Potential in Hyporheic Zone Sediment Columns.

Hyporheic zones (HZ) are active biogeochemical regions where groundwater and surface water mix. N transformations in HZ sediments were investigated in columns with a focus on understanding how the dynamic changes in groundwater and surface water mixing affect microbial community and its biogeochemical function with respect to N transformations. The results indicated that denitrification, DNRA, and nitrification rates and products changed quickly in response to changes in water and sediment chemistry, fluid residence time, and groundwater-surface water exchange. These changes were accompanied by the zonation of denitrification functional genes along a 30 cm advective flow path after a total of 6 days' elution of synthetic groundwater with fluid residence time >9.8 h. The shift of microbial functional potential toward denitrification was correlated with rapid NO3- reduction collectively affected by NO3- concentration and fluid residence time, and was resistant to short-term groundwater-surface water exchange on a daily basis. The results implied that variations in microbial functional potential and associated biogeochemical reactions in the HZ may occur at space scales where steep concentration gradients present along the flow path and the variations would respond to dynamic HZ water exchange over different time periods common to natural and managed riverine systems.

Polymerase chain reaction-hybridization method using urease gene sequences for high-throughput Ureaplasma urealyticum and Ureaplasma parvum detection and differentiation.

In this article, we discuss the polymerase chain reaction (PCR)-hybridization assay that we developed for high-throughput simultaneous detection and differentiation of Ureaplasma urealyticum and Ureaplasma parvum using one set of primers and two specific DNA probes based on urease gene nucleotide sequence differences. First, U. urealyticum and U. parvum DNA samples were specifically amplified using one set of biotin-labeled primers. Furthermore, amine-modified DNA probes, which can specifically react with U. urealyticum or U. parvum DNA, were covalently immobilized to a DNA-BIND plate surface. The plate was then incubated with the PCR products to facilitate sequence-specific DNA binding. Horseradish peroxidase-streptavidin conjugation and a colorimetric assay were used. Based on the results, the PCR-hybridization assay we developed can specifically differentiate U. urealyticum and U. parvum with high sensitivity (95%) compared with cultivation (72.5%). Hence, this study demonstrates a new method for high-throughput simultaneous differentiation and detection of U. urealyticum and U. parvum with high sensitivity. Based on these observations, the PCR-hybridization assay developed in this study is ideal for detecting and discriminating U. urealyticum and U. parvum in clinical applications.

Identification of a novel mitochondrial interacting protein of C1QBP using subcellular fractionation coupled with CoIP-MS.

The study of protein-protein interactions is an essential process to understand the biological functions of proteins and the underlying mechanisms. Co-immunoprecipitation coupled with mass spectrometry (CoIP-MS) is one of the most extensively used high-throughput techniques to discover novel protein-protein interactions. However, the traditional CoIP process uses whole cell lysate, disrupts cellular organization, and leads to potential false positives by inducing artificial protein-protein interactions. Here, we have developed a strategy by combining subcellular fractionation with CoIP-MS to study the interacting proteins of the complement component 1, q subcomponent binding protein (C1QBP) in the mitochondria. Using this method, a novel C1QBP interacting protein, dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex, mitochondrial (DLAT) was identified and validated. Furthermore, the activity of the pyruvate dehydrogenase (PDH) was found to be affected by the expression level of C1QBP. These results provide novel insights regarding the mitochondrial function of C1QBP in the regulation of cellular energy metabolism. This method could also be used to analyze the subcellular protein-protein interactions for other proteins of interest.

Methods Comparison and Bias Estimation of Three Distinct Biochemistry Analytical Systems in One Clinical Laboratory Using Patient Samples.

BACKGROUND: In China, one laboratory usually owns more than one diagnostic device or reagent kit measuring the same analyte and this situation causes great troubles for quality control and traceability. To determine if the different devices yield equivalent results, the correlation coefficients and predicted bias between three distinct bio-chemistry analyzers in our lab were evaluated. METHODS: 40 analytes were detected and used to evaluate method performance and comparability of results between different analyzers. The Vitros5600 and Hitachi7170 analyzers were separately compared with the Cobas 8000 analyzer according to Clinical and Laboratory Standards Institute (CLSI) guideline (EP9-A2). Within-day and between-day imprecision of the three analytical systems were calculated in accordance with CLSI guidelines EP15-A2. RESULTS: Comparing the Hitachi7170 with Cobas8000 analyzer, except for calcium, magnesium, chloride ion (CL-), and carbon dioxide, the other 36 analytes were closely correlated (R2 > 0.95), while 4 of the 36 analytes' predicted bias exceeded the acceptable criteria. As for the Vitros5600 and Cobas8000, except for albumin, sodium ion (NA+), magnesium, and chloride (CL), the other 13 analytes were closely correlated (R2 > 0.95), while 5 of the 13 analytes' predicted bias exceeded the acceptable criteria. CONCLUSIONS: Significant differences for several analytes between distinct analyzers were found; for some analytes the predicted bias between dry chemical analyzer and conventional wet chemical analyzer can reach 30%, which is worthy of our concern. When one analyte was detected on more than one device, a strict method comparison study should be performed regularly. Reference intervals should be validated and transferred from the Cobas 8000 to Vitros5600 when the bias cannot be adjusted.

Corin is down-regulated and exerts cardioprotective action via activating pro-atrial natriuretic peptide pathway in diabetic cardiomyopathy.

BACKGROUND: Diabetic cardiomyopathy (DCM), a fatal cardiovascular complication of diabetes mellitus, often leads to progressive heart failure, however its pathogenesis remains unclear. Corin, a cardiac serine protease, is responsible for converting pro-atrial natriuretic peptide (pro-ANP) to biologically active atrial natriuretic peptide (ANP). It has been well established that corin deficiency is associated with the progression of hypertension, cardiac hypertrophy and heart failure. However, because the involvement of corin-mediated pro-ANP processing in DCM has not been clarified, this study aims to investigate the role of corin in the pathogenesis of DCM. METHODS: Diabetes mellitus was induced by a single intraperitoneal injection of streptozotocin (STZ 65 mg/kg) to Sprague-Dawley rats (180-220 g). DCM was confirmed by monitoring continuously transthoracic echocardiography every 4 weeks and hemodynamic measurements at 20 weeks. Myocardial disorder and fibrosis were detected by HE staining and Masson's trichrome staining. The mRNA and protein levels of corin and ANP in rat hearts and cardiomyocytes were determined by quantitative real-time PCR, western blotting and immunohistochemical staining, respectively. H9c2 cardiomyoblasts proliferation was detected by MTT colorimetric assay and viable cell counting with trypan blue. The effect of Corin-siRNA H9c2 cardiomyoblasts on EA.hy926 cells migration was measured by the wound healing scratch assay. RESULTS: The corin and ANP expression in mRNA and protein levels was decreased in DCM rat hearts. Corin and ANP levels of neonatal rat cardiomyocytes and H9c2 cardiomyoblasts treated with high glucose were significantly lower than that of normal glucose treated. Precisely, corin and ANP levels decreased in DCM rats at 12, 16, 20 and 33 weeks; neonatal cardiomyocytes and H9c2 cardiomyoblasts treated with high glucose at 36, 48 and 60 h demonstrated significant reduction in corin and ANP levels. Corin-siRNA H9c2 cardiomyoblasts showed decreased proliferation. Culture supernatants of Corin-siRNA H9c2 cardiomyoblasts prevented endothelial cell line EA.hy926 migration in the wound healing scratch assay. Furthermore, iso-lectin expression in arteriole and capillary endothelium was down-regulated in DCM rats. CONCLUSIONS: Our results indicate that corin plays an important role in cardioprotection by activating pro-atrial natriuretic peptide pathway in DCM. Corin deficiency leads to endothelial dysfunction and vascular remodeling.

Overexpression of YB1 and EZH2 are associated with cancer metastasis and poor prognosis in renal cell carcinomas.

Renal cell carcinoma (RCC) is the most common type of kidney cancers in adults, and metastasis represents the major cause of mortality of RCC patients. The Y-box binding protein 1 (YB1) is a multifunctional oncoprotein in various malignancies. Enhancer of zeste homolog 2 (EZH2), a polycomb histone methyltransferase, is a key epigenetic modifier implicated in various cancer metastasis. However, the expression patterns and clinical correlations of both YB1 and EZH2 in RCC remain largely unclear. In this study, the expression of YB1 and EZH2 were examined using immunohistochemistry staining in a study cohort including 165 RCC and 80 tumor adjacent normal tissues. RCC tissues showed a significant higher nuclear expression of YB1 (p < 0.001) and EZH2 (p < 0.001) as compared with the normal counterparts. In addition, YB1 and EZH2 nuclear overexpression were found to be positively associated with RCC stage (p < 0.001 and p = 0.005), Fuhrman tumor grade (p = 0.022 and p = 0.044), and metastasis (p < 0.001 and p = 0.009). Overall survival analysis indicated patients with YB1 (p = 0.004, HR 5.656 (2.006-10.944)) and/or EZH2 (p = 0.006, HR 4.551 (2.124-9.438)) nuclear overexpression correlated with poor survival. More interestingly, YB1 and EZH2 nuclear expression was correlated (p = 0.005). Further studies demonstrated that EZH2 expression was significantly downregulated in YB1 knockdown RCC cell lines. Functionally, YB1 knockdown inhibited RCC invasion in vitro. In conclusion, YB1 and EZH2 expression was correlated and associated with RCC incidence, tumor stage, grade, metastasis, and survival.

Elevated Plasma P-Selectin Autoantibodies in Primary Sjögren Syndrome Patients with Thrombocytopenia.

BACKGROUND Primary Sjögren's syndrome (pSS) is one of the most common chronic systemic autoimmune diseases, and thrombocytopenia is one of the hematological manifestations of pSS. When platelet and endothelial cells are activated, P-selectin is expressed on the cell surface. This study aimed to investigate the role of P-selectin autoantibodies in the pathogenesis of thrombocytopenia in pSS. MATERIAL AND METHODS P-selectin autoantibodies were measured by enzyme-linked immunosorbent assay (ELISA) in 38 pSS patients without thrombocytopenia and 32 pSS patients with thrombocytopenia, 32 idiopathic thrombocytopenic purpura (ITP) patients, and 35 healthy controls. RESULTS The plasma P-selectin autoantibodies (A490) in ITP patients and pSS patients with/without thrombocytopenia were significantly higher than those in healthy controls, but there were no significant differences between ITP patients and pSS patients with thrombocytopenia. The positive rate of P-selectin autoantibodies in pSS patients with thrombocytopenia was significantly higher than that in ITP patients. The platelet count was lower in P-selectin autoantibodies-positive patients, while among pSS patients with thrombocytopenia, the platelet count was lower in P-selectin autoantibodies-positive patients than in P-selectin autoantibodies-negative patients. In ITP patients and pSS patients with thrombocytopenia, the platelet count was lower in P-selectin autoantibodies-positive patients. CONCLUSIONS Elevated plasma P-selectin autoantibodies may play a role in the pathogenesis of thrombocytopenia in pSS patients.

MiR-26a enhances metastasis potential of lung cancer cells via AKT pathway by targeting PTEN.

Lung cancer is the leading cause of cancer related death, 90% of lung cancer patients die of metastasis. Many microRNAs (miRNAs) are deregulated in cancer. They are involved in tumorigenesis and function as oncogenes or tumor suppressor genes. Recent studies show that miRNAs may be responsible for tumor metastasis. Several functional studies show that miR-26a plays an important role in carcinogenesis; however, none of these studies is related to tumor metastasis. In the present study, we investigated the effect of miR-26a on metastasis potential of lung cancer cells. Our data showed that miR-26a expression level was higher in lymph node metastasis tumor tissues than in primary tumor tissues. Ectopic expression of miR-26a dramatically enhanced lung cancer cell migration and invasion abilities. Metastasis-related genes matrix metallopeptidase 2 (MMP-2), vascular endothelial growth factor (VEGF), Twist and ß-catenin were upregulated. Phosphatase and tensin homolog (PTEN) was a direct target of miR-26a. Further mechanistic study revealed that miR-26a increased AKT phosphorylation and nuclear factor kappa B (NFκB) transcriptional activation. Our study demonstrated that miR-26a enhanced lung cancer cell metastasis potential via modulation of metastasis-related gene expression, and activation of AKT pathway by PTEN suppression, suggesting that miR-26a might be a potential therapeutic candidate in patients with metastatic lung cancer.

Critical role of TRPC6 channels in the development of human renal cell carcinoma.

Renal cell carcinoma (RCC) is the most common tumor arising from the cells in the lining of tubules in the kidney. Some members of the Ca2+-permeable transient receptor potential canonical (TRPC) family of channel proteins have demonstrated a role in the proliferation of some types of cancer cells. In this study, we investigated the role of TRPC6 in the development of human RCC. RT-PCR and Western blotting were used to investigate TRPC6 expression in 1932 and ACHN cells. Immunohistochemical techniques were applied to study TRPC6 expression in 60 cases of RCC primary tissue samples and 10 cases of corresponding normal renal tissues. To inhibit TRPC6 activity or expression, RNA interference was used. The effects of TRPC6 channels on RCC cell viability and cell cycle progression were investigated by MTT and flow cytometry. TRPC6 was expressed in 1932 and ACHN cells. TRPC6 protein was detected in 73.3% of RCC samples, and there was a significant difference compared with the normal renal samples (30%) (p<0.05). Moreover the level of TRPC6 expression was associated with RCC Fuhrman grade (p<0.01). Blockade of TRPC6 channels in ACHN cells suppressed basal cell proliferation and partially inhibited HGF-induced cell proliferation. Furthermore, inhibition of TRPC6 channels expression prolonged the transition through G2/M phase in ACHN cells. In summary, expression of TRPC6 is markedly increased in RCC specimens and associated with RCC histological grade. TRPC6 plays an important role in ACHN cells proliferation.

Drug-drug interactions between ketoconazole and berberine in rats: pharmacokinetic effects benefit pharmacodynamic synergism.

Ketoconazole (KTZ), a clinical antifungal agent, is a known inhibitor of CYP3A and permeability glycoprotein (P-gp). Berberine (BBR), a natural plant-derived product used for gastroenteritis, is a substrate of P-gp. Recently, a synergistic antifungal effect of KTZ combined with BBR has been revealed. In this study, we performed both in vivo and in vitro experiments to explore whether pharmacokinetic interactions between KTZ and BBR would benefit their pharmacodynamic synergism. After oral co-administration of 10 mg/kg KTZ with 60 mg/kg BBR, the average area under the curve (AUC) and the maximum concentration (C(max) ) for KTZ increased to 215% and 449% (p < 0.05), respectively, in male rats and 157% and 172% (p < 0.05), respectively, in female rats, compared with those administered KTZ alone. Area under the curve and C(max) for BBR increased to 173% and 142%, respectively, compared with those administered BBR alone. After intravenous co-administration of 0.5 mg/kg KTZ and 0.8 mg/kg BBR, the pharmacokinetic properties of KTZ remained the same, but AUC of BBR increased to 254% (p < 0.05) compared with those administered BBR alone. In rat liver microsomes, inhibitory concentration (IC)(50) of BBR inhibiting KTZ depletion was determined to be 103 µm. These resulting pharmacokinetic interactions may benefit their pharmacodynamic synergism to a certain extent.

Source and enrichment mechanism of fluoride in groundwater of the Hotan Oasis within the Tarim Basin, Northwestern China.

In arid inland irrigated areas, the role of human activities on fluoride enrichment in groundwater is not fully understood. There is an extremely arid climate, high-intensity irrigation, and severe soil salinization in the Hotan Oasis within the Tarim Basin, Northwestern China. In this study, hydrogeochemistry and environmental isotope methods were combined to explore the distribution characteristics and controlling processes of fluoride enrichment in groundwater. The F- concentration in groundwater had a range of 1.12-9.4 mg/L. F- concentrations of all the groundwater samples were higher than 1.0 mg/L (Chinese Standards for Drinking Water Quality), and about 89% were higher than 1.5 mg/L (WHO Guidelines for Drinking Water Quality). High fluoride groundwater was mainly distributed downstream of the river and in the middle of the interfluvial zone. Vertically, the fluoride concentration was higher when the sampling depth was less than 15 m. There was a significant positive correlation between F- concentration and salinity in groundwater. F- in groundwater was mainly derived from river water fluoride, which could be imported to groundwater with infiltration of rivers and irrigation canals as well as irrigation return flow. Anthropogenic inputs may be partly responsible for fluoride enrichment in groundwater. Fluoride accumulated in the vadose zone by strong evapotranspiration and then leached into groundwater with irrigation return flow was the main mechanism of F- enrichment in groundwater in the study area. This work is a clear example of how human activities together with natural processes can affect the chemical quality of groundwater, which is essential to safeguard the sustainable management of water and soil resources inland arid oasis areas.

Insights into carbon isotope fractionation on trichloroethene degradation in base activated persulfate process: The role of multiple reactive oxygen species.

Understanding the role of reactive oxygen species (ROS) is essential to elucidate the mechanism of contaminants degradation in in-situ chemical oxidation (ISCO). In this study, compound specific isotope analysis (CSIA) and radicals quenching methods were integrated to investigate the roles of hydroxyl radical (HO), sulfate radical (SO4-), and superoxide radical (O2-) on trichloroethene (TCE) degradation during persulfate (PS) activated with base. The carbon isotope fractionation of TCE was found to be dependent of the base:PS ratios, yielding carbon enrichment factors (ε values) from -9.8 ± 0.5‰ to -16.7 ± 1.0‰ at base:PS molar ratios between 0.5:1 and 10:1, which was attributed to multi-pathways degradation of TCE by multiple ROS. The expected ε value (-31.6 ± 1.6‰) for TCE degradation via O2- attacking pathway, was more negative than those values via SO4- or HO pathways. The relative contributions of HO, SO4- and O2- for TCE degradation during base activated PS were estimated with observed ε values. HO and O2- were the predominant ROS for TCE degradation (with the relative contribution of 55-69% and 22-45%, respectively) in base activated PS. This work highlights the prospect of CSIA application for identifying degradation pathways of contaminants with ROS in environment.

Identification of groundwater pollution from livestock farming using fluorescence spectroscopy coupled with multivariate statistical methods.

Extensive livestock farming has highly threatened groundwater quality, thereby necessitating a rapid and effective method to identify groundwater quality in such areas. Fluorescence spectroscopy has been recognized as an interpretable method for tracking anthropogenic influences on water quality, but its applicability in identifying the groundwater pollution from livestock farming remains unknown. In this study, the fluorescence characteristics of dissolved organic matter (DOM) in groundwater from a typical livestock farming area were investigated by using fluorescence excitation emission matrix (EEM)-parallel factor analysis (PARAFAC) coupled with multivariate statistical methods. The results showed that livestock farming significantly altered the content and composition of DOM in groundwater, and these effects were mainly observed in shallow groundwater in the study area. Hierarchical cluster analysis based on fluorescence parameters divided the groundwater samples into three clusters with significantly different pollution degrees: Cluster A, unpolluted; Cluster B, highly polluted; Cluster C, moderately polluted. In particular, the intensity of tryptophan-like fluorescence was high in the polluted groundwater but was almost undetectable in the unpolluted groundwater, suggesting that it is a potential indicator of groundwater quality. Principal component analysis based on the fluorescence parameters explained 91.5% of the variance with the first two principal components, and revealed that the degree of pollution dominated the fluorescence characteristics of groundwater in the study area. In addition, NO3- was abundant in Clusters B and C, while it was low in Cluster A, validating the analysis results of fluorescence spectroscopy. These findings indicated that DOM fluorescence was sensitive to livestock farming pollution and could be applied to identify, monitor, and assess groundwater pollution from livestock farming.

Hydrogeochemistry Evidence for Impacts of Chemical Acidic Wastewater on Karst Aquifer in Dawu Water Source Area, Northern China.

The study of the hydrochemical characteristics and the water-rock interaction of karst groundwater is very important for the rational exploitation of karst groundwater and its pollution control. In this paper, the systematic clustering method was used to analyze the hydrochemical characteristics of different types of groundwater, combined with hydrochemical graphic analysis and correlation analysis to explore the impact of chemical acidic wastewater on the evolution of karst aquifer in the Dawu water source area, northern China. The results show that the chemical acid wastewater, sourcing from discharges/spillages from the local chemical industries, has different degrees of pollution impact on karst groundwater, causing the total hardness of all karst groundwater and the total dissolved solids, Cl- and SO42- in nearly half of the karst groundwater to exceed the quality indexes of class III water in China's standard for groundwater quality (GB/T 14848-2017). Hydrochloric acid and sulfuric acid in the wastewater can be buffered by the dissolution of carbonate rocks, resulting in a nearly neutral pH (pH-buffering effect) and an increase in Ca2+, Mg2+, Sr, Cl- and SO42- concentrations in karst groundwater.

Free CA125 promotes ovarian cancer cell migration and tumor metastasis by binding Mesothelin to reduce DKK1 expression and activate the SGK3/FOXO3 pathway.

Objective: CA125/MUC16 is an O-glycosylated protein that is expressed on the surfaces of ovarian epithelial cells. This molecule is a widely used tumor-associated marker for diagnosis of ovarian cancer. Recently, CA125 was shown to be involved in ovarian cancer metastasis. The purpose of this study was to investigate the mechanism of CA125 during ovarian cancer metastasis. Methods: We analyzed the Oncomine and CSIOVDB databases to determine the expression levels of DKK1 in ovarian cancer. DKK1 expression levels were upregulated or downregulated and applied with CA125 to Transwell and Western blot assays to ascertain the underlying mechanism by which CA125 stimulates cell migration via the SGK3/FOXO3 pathway. Anti-mesothelin antibodies (anti-MSLN) were used to block CA125 stimulation. Then the expression levels of DKK1were tested by enzyme-linked immunosorbent assay (ELISA) to eliminate the blocking effect of anti-MSLN to CA125 stimulation. Xenograft mouse models were used to detect the effects of CA125 and anti-MSLN on ovarian cancer metastasis in vivo. Results: DKK1 levels were downregulated in ovarian tumor tissues according to the analyses of two databases and significantly correlated with FIGO stage, grade and disease-free survival in ovarian cancer patients. DKK1 levels were downregulated by CA125 stimulation in vitro. Overexpression of DKK1 reversed the ability of exogenous CA125 to mediate cell migration by activating the SGK3/FOXO3 signaling pathway. Anti-MSLN abrogated the DKK1 reduction and increased the apoptosis of ovarian cancer cells. The use of anti-MSLN in xenograft mouse models significantly reduced tumor growth and metastasis accelerated by CA125. Conclusions: These experiments revealed that the SGK3/FOXO3 pathway was activated, wherein decreased expression of DKK1 was caused by CA125, which fuels ovarian cancer cell migration. Mesothelin is a potential therapeutic target for the treatment of ovarian cancer metastasis.

Shallow Groundwater Quality and Its Controlling Factors in the Su-Xi-Chang Region, Eastern China.

Understanding factors influencing groundwater quality is critical to the development of best management practices at the large watershed scale. In this study, the shallow groundwater (10-20 m depth) in the Su-Xi-Chang region, eastern China, was investigated as part of a monitoring program from 2007 to 2008 to analyze the regional groundwater quality as well as the hydrogeochemical processes and their controlling factors. Conventional physicochemical water parameters (pH, turbidity, electrical conductivity, dissolved oxygen, total phosphorus), major cations (Na+, Ca2+, Mg2+ and NH4+) and anions (Cl-, NO3- and SO42-) were measured. Hydrochemical methods and multivariate statistical methods were applied to analyze the hydrogeochemical signatures, origins, the similarities among the variables and to identify the main pollution sources in the groundwater. The results showed that (1) the concentrations of TDS (224.89-1086.70 mg/L) and turbidity (0.1-18.60 NTU) were higher than the class II groundwater quality standards in China and the WHO drinking water standards, (2) there were extremely high concentrations of ammonia (0.01-32.90 mg/L), with a mean value of 0.72 mg/L and (3) the nitrate concentrations (average value of 22.07 mg/L) exceeded the class III groundwater quality standards. The study also provided evidence that weathering, dissolution of carbonate, halite and silicate and cation exchange were the possible primary hydrogeochemical control mechanisms in the groundwater. The sources of ammonia, total phosphorus, sulfates and nitrates included rock-water interactions and anthropogenic activities. The groundwater administration of pollution sinks and sources, long-term legal frameworks and economic incentives should be improved to optimize watershed scale management in the context of rapid development in China.