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1.
Methods Mol Biol ; 2050: 13-19, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31468475

RESUMO

Generally, electroporation of in vitro cells is performed under very high electric fields to overcome the physical barrier of plasma membrane. Since traditional electroporation techniques make use of very high voltages, which is critical to cell viability, this study presents a microfluidic platform able to perform cell membrane electroporation with the application of low voltages (1.5-2 V). The platform is manufactured based on the milling by mean of focused ionic beam, which offers an established approach to fabricate ordered arrays of 3D gold hollow nanoelectrodes protruding from an insulating substrate. The novelty of this fabrication relies on the fact that the nanoelectrodes used for electroporation are simultaneously metallic, hollow and communicate through its nanochannels with an isolated microfluidic chamber beneath the device. Adherent cultured cells on the nanoelectrodes can be electroporated in this platform, and molecules can be selectively delivered only inside the porated cells.

2.
J Nanosci Nanotechnol ; 20(3): 1859-1865, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31492353

RESUMO

As oxidation is inevitable during application of microwave absorbing materials (MAM) exposed to oxygen-containing atmosphere at elevated temperatures. The investigations concerning influences of oxidation on microstructure and microwave absorption (MA) properties are of great significance. Aiming to better regulating MA for Si3N4-based ceramics serviced in high temperature environment, the evolution of MA properties of SiCw/Si3N4 composites (SwSN) after oxidation at 1200 °C in ambient air for short time was studied. It was found that after oxidation for 7.5 h, both the real and imaginary permittivity of SwSN showed only slight decrease due to good oxidation protection provided by dense SiO2 layer formed on the surface. Moreover, SwSN after short-time of oxidation also showed an almost negligible decline in MA properties. SwSN which has good oxidation resistance and stable microwave attenuation abilities was subjected to oxidation at 1200 °C for short time. The results showed its favorable and potential applications in high-temperature microwave absorption fields.

3.
Methods Mol Biol ; 2057: 103-112, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31595474

RESUMO

Nitrogen (N2) is the most necessary element in the atmosphere, it is an energetic micronutrient for plant growth and development after water, besides its key role in chlorophyll production, which is crucial for photosynthesis process. Biological nitrogen fixation is measured to be the most potent method to deliver a fixed way of nitrogen to the plants. Plant depends on free-living and symbiotic microbes present in the soil for nitrogen because it cannot be absorbed by the plant itself directly from the atmosphere. Many techniques were reported in the laboratory for nitrogen estimation till now, but Kjeldahl digestion and acetylene reduction assay (ARA) techniques became the most popular. In this chapter, we focus on the most common and popular methods used to determine plant N2; awareness obtained through the wide application of these methods should offer the source for the N2 fixation rate in agriculture system.

4.
Artigo em Inglês | MEDLINE | ID: mdl-31568155

RESUMO

BACKGROUND/RATIONALE: Biliary atresia (BA) is a cholangiopathy characterized by bile flow obstruction due to destruction of the biliary tree. Without surgical correction with Kasai portoenterostomy (KPE), BA leads to death or liver transplant (LTx). Early-onset, progressive liver fibrosis is a defining characteristic of BA. Collagen hybridizing peptide (CHP) is a synthetic peptide which binds to denatured collagen strands allowing quantification of fibrosis. This technique has not been used on human liver tissue. The aim of this pilot study was to evaluate the utility of CHP as a measurement of quantitative fibrosis to allow earlier survival with native liver (SNL) prognostication. RESULTS: We identified 21 patients with wedge liver biopsies available, of which 14 required LTx. No deaths occurred. Patients requiring LTx tended to be female with a significantly different mean bilirubin (p = 0.002), albumin (p = 0.001) and ALT (p = 0.03) at 3-months post-KPE. By 1-year post-KPE, 50% of patients in the high-CHP intensity group required LTx versus 27% in the low-CHP. Overall, fibrosis as quantified by CHP at time of KPE was associated with more than three-times the risk of requiring LTx by 4-years of age (HR 3.6, 95%CI 1.15-10.93, p = 0.03). When controlling for gender and TB > 2 mg/dL and albumin at 3-months post-KPE, it predicted nearly seven times the risk of LTx (HR 6.89, 95%CI 1.38-34.32, p = 0.02). CONCLUSION: Our results suggest that quantitative assessment of fibrosis at the time of KPE holds promise as an earlier predictor of LTx requirement in BA. A larger study is justified to assess quantitative fibrosis as a BA prognostic tool.

5.
Vasc Endovascular Surg ; : 1538574419879849, 2019 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-31597540

RESUMO

BACKGROUND: The carotid sheath contains clinically important and vital anatomical neurogenic, vascular, and lymphatic structures that allow for a great variety of lesions. Vascular anomalies found in the carotid sheath are rarely reported and may be easily misdiagnosed as arterial aneurysms, neurogenic tumors, paragangliomas, or lymphatic masses. METHOD: We present a 60-year-old woman with a vascular malformation arising within the carotid sheath at the right carotid bifurcation, which mimics carotid body tumor. RESULT: The mass was excised successfully with an uneventful postoperative course, and histological analysis suggested a vascular malformation with thin-walled blood vessels, lined by endothelial cells and separated by fibrous tissue. The patient was in good clinical condition without signs of relapse of the mass at 6-month follow-up. CONCLUSION: Vascular malformation is a rare but existing possibility of pathology in the carotid sheath, which can be effectively removed by meticulous surgery.

6.
Ren Fail ; 41(1): 946-953, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31599192

RESUMO

Objective: Despite the achievement of blood glucose, blood pressure targets, the risk for kidney injury remains high among older adults. This observational retrospective study investigated whether high TG or high WC contribute to this high residual risk for kidney injury. Methods: A total of 843 elderly from Dongli Community, Tianjin, China, we selected 666 individuals with a baseline estimated glomerular filtration rate (eGFR) ≥60 mL/min/1.73 m2 and negative microalbuminuria completing a 3-year follow-up. At baseline, subjects were grouped according to the levels of TG and WC. The primary outcome was the incidence of kidney injury, defined as low eGFR (eGFR <60 mL/min/1.73 m2) or reduced eGFR (eGFR reduced >25%) or UACR ≥30 mg/g. Results: Overall, 6.01% developed low eGFR, 11.11% reduced eGFR, 25.98% UACR ≥30 mg/g, and 3.45% low eGFR and UACR ≥30mg/g after 3-year follow-up. TG ≥1.7 mmol/L increased the risk of eGFR <60 mL/min/1.73 m2 by 1.44-fold, of UACR ≥30 mg/g by 32%, and of developing both abnormality by 1.41-fold in model 1; further adjustment for potential confounders factors, the association is slightly weakened in model 2 and 3; WC (≥90 cm in men and ≥85 cm in women) were associated with a 1.68-fold higher risk of eGFR <60 mL/min/1.73 m2 and a 1.43-fold risk of UACR ≥30 mg/g and a 1.89-fold risk of developing both abnormality in model 1. Further adjustment for potential confounders factors, the association is slightly weakened in model 2 and 3. Conclusions: In a population of Chinese community-dwelling older adults, high TG and central obesity were risk factors for the development of kidney injury over 3 years.

7.
Artigo em Inglês | MEDLINE | ID: mdl-31593347

RESUMO

Here, high-oriented mesoporous graphitic carbon nanospring (OGCS) with graphitic layers that are perpendicular to the axis is prepared by hydrothermal treatment of epoxy resin at 500 ℃ and followed by annealing at 1400 ℃. It is demonstrated that the water plays an important role in not only forming graphitic carbon nanospring with high [002] orientation and a large amount of active edge-plane sites, but also contributing to the generation of mesoporous structure, which both facilitate fast K-ion adsorption and diffusion. On basis of in situ and ex situ measurements, it is confirmed that OGCS undergoes K-adsorption in mesopores at first and then K-intercalation in graphite layer to form KC 8 with low voltage. Besides, the spring-like nanostructure can expand one-dimensionally along the axial direction to accommodate volume variation. As a result, the synthesized OGCS electrode shows a much better K-storage performance than that of unoriented graphitic carbon, such as a stable capacity of 361.7 mA h g -1 at 50 mA g -1 over 100 cycles, good rate capability with capacity of 170 mA h g -1 even at 2000 mA g -1 , and long-term cycling stability over 10000 cycles.

8.
Int J Nanomedicine ; 14: 7237-7247, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31564876

RESUMO

Background: The health hazards of silica nanoparticle (SiNP) are raising worldwide concern as SiNPs has become the second largest manufactured nanomaterial in global markets. However, insufficient data for the adverse health effects and safety evaluation of SiNPs are remaining a big question. Purpose: We evaluated the effects and related mechanism of SiNPs on pulmonary inflammation and collagen production through repeated intravenous administration in mice in a 45-day observation period. Methods: Morphological and ultrastructural change, ultradistribution of SiNPs in lungs were observed in ICR mice through intravenous administration. Oxidative damage, pro-inflammatory cytokines, hydroxyproline content, the marker of fibroblasts and epithelial-mesenchymal transition (EMT), and JAK2/STAT3 and TGF-ß1/Smad3 signaling pathways were detected to explore the lung injuries and related mechanism. Results: The results showed repeated intravenous exposure of SiNPs increased the weight of lung tissues and destroyed pulmonary histomorphological structure. The increased MDA content, depletion of SOD and GSH-Px in lungs were observed in SiNP-treated mice. The protein expressions of JAK2/STAT3 pathway were upregulated in lungs, and the levels of inflammatory cytokines TNF-α, IL-1ß, and IL-6 in serum and lungs were also elevated in SiNPs treated group. The increased hydroxyproline content indicated collagen accumulation in lungs of SiNP-treated mice. Meanwhile, the protein expressions of the marker of myofibroblast (a-SMA), the regulators in connective tissue remodeling (CTGF), TGF-ß, and p-Smad3 were all upregulated in lungs. In addition, we found intravenous administration of SiNPs-induced ultrastructural changes in type II alveolar epithelial cells but without downregulation of the protein expression of the key markers of epithelial cells (E-Cadherin). Conclusion: Our results revealed that oxidative stress and inflammation contributed to the collagen accumulation through activation of JAK2/STAT3 and TGF-ß/Smad3 pathways. It suggests that pulmonary aberrant inflammation and collagen accumulation induced by nanoparticles should be seriously considered for the safety application in diagnostics or therapeutics.

9.
Chem Commun (Camb) ; 2019 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-31588453

RESUMO

Hollow PdO-Co3O4 nano-dodecahedrons with moderate catalytic activity were designed as electrocatalysts for Li-O2 batteries, and not only reduce the overpotentials effectively but also improve the reversibility of the ORR and OER, and cycle for more than 90 cycles stably with extremely low overpotentials.

10.
Sci Rep ; 9(1): 14118, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31575964

RESUMO

Chronic hepatitis B virus (HBV) infection may follow four different consecutive phases, which are defined by virology as well as biochemical markers and differ in terms of prognosis and need for antiviral treatment. Currently, host responses reflected by immune markers are not considered in this definition. We aimed to study soluble immune markers and their distribution in different phases of chronic HBV infection. In this cross-sectional retrospective study, we investigated a panel of 14 soluble immune markers (SIM) including CXCL10 in 333 patients with chronic HBV infection. In a small cohort of HBeAg positive patients we analyzed SIM before and after HBeAg seroconversion and compared seroconverters to patients with unknown outcome. Significant differences were documented in the levels of several SIM between the four phases of chronic HBV infection. The most pronounced difference among all investigated SIM was observed for CXCL10 concentrations with highest levels in patients with hepatitis. TGF-ß and IL-17 revealed different levels between HBeAg negative patients. HBeAg positive patients with HBeAg seroconversion presented higher amounts of IL-12 before seroconversion compared to HBeAg positive patients with unknown follow up. SIM such as CXCL10 but also IL-12, TGF-ß and IL-17 may be useful markers to further characterize the phase of chronic HBV infection.

11.
Artigo em Inglês | MEDLINE | ID: mdl-31589799

RESUMO

A series of supramolecular assemblies of types [Ag 8 L 4 ](PF 6 ) 8 and [Ag 4 L 2 ](PF 6 ) 4 obtained from tetraphenylethylene (TPE) bridged tetrakis(1,2,4-triazolium) salts H 4 -L(PF 6 ) 4 and Ag I ions is described. The type of assembly obtained is highly dependent on the N-wingtip substituents of the tetrakis(1,2,4-triazolium) starting materials (H 4 -L)(PF 6 ) 4 . Minute changes in the lengths of the N4-wingtip substituents enables the controlled formation of assemblies with either [Ag 4 L 2 ](PF 6 ) 4 or [Ag 8 L 4 ](PF 6 ) 8 stoichiometry. The molecular structures of selected [Ag 8 L 4 ](PF 6 ) 8 and [Ag 4 L 2 ](PF 6 ) 4 assemblies have been determined by X-ray diffraction analyses. While the tetrakis(1,2,4-triazolium) salts do not exhibit fluorescence in solution, all of their tetra-NHC assemblies do due to rigidification of the TPE core upon NHC-metal coordination. Upon irradiation, all assemblies undergo a light-induced, supramolecule-to-supramolecule structural transformation via an oxidative photocyclization involving phenyl groups of the TPE core, resulting in a significant change of the luminescence properties.

12.
Artigo em Inglês | MEDLINE | ID: mdl-31566350

RESUMO

The comfort and protection of clothes are critically important for human well-being in life; however, constructing multifunctional fabrics with excellent thermoregulating and water-repellent performance still presents an exciting scientific challenge and a significant technological advancement. Therefore, we report a novel and straightforward methodology to fabricate corncoblike and phase-changeable nanofibers by incorporating n-octadecane phase change capsules (PCCs) for creating water-repellent and thermoregulating nanofibrous membranes. This strategy causes PCC to be uniformly distributed on the nanofibers to form a unique corncoblike structure, preventing the abscission of PCC and the leakage of the phase change ingredient (n-octadecane). Besides, the resultant nanofibrous membranes are endowed with hierarchical roughness, small pore size, and energy storage/release capacity in response to environmental changes. As a consequence, the nanofibrous membranes present prominent water repellence with superhydrophobicity (a water contact angle of 153°) and a high hydrostatic pressure of 84 kPa, robust mechanical properties with a tensile strength of 9.2 MPa, as well as excellent hybrid active-passive thermal regulation with a water vapor transmittance rate of 11.4 kg m-2 d-1 and a high phase change enthalpy of 74 J g-1 after 50 heating/cooling cycles, indicating them to be an exceptional candidate for personal protection and thermal management.

13.
J Am Chem Soc ; 2019 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-31600069

RESUMO

A diverse collection of copper-catalyzed intermolecular aminative difunctionalizations of unactivated alkenes with N-halodialkylamines as the terminal dialkylamino source is reported. A bidentate auxiliary tethered on the alkene substrates is crucial, which can promote the migratory insertion of nonactivated alkenes into the aminyl radical-metal complex and stabilize the resultant high-valent copper intermediate to allow for further transformations. By employing this strategy, the intermolecular aminohalogenation reactions and a three-component aminoazidation reaction of unactivated alkenes with dialkylamino source were successively achieved in a remarkable regio- and stereoselective manner. These reactions were performed under neutral conditions and maintained excellent functional group tolerance toward a wide range of N-halodialkylamines and unactivated alkenes. Further mechanistic studies and DFT calculations supported a concerted migratory insertion of the C-C double bond into the aminyl radical-metal complex to form a Cu(III) intermediate.

14.
Eur Respir J ; 2019 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-31601711

RESUMO

WHO (World Health Organization) recommends countries to implement pharmacovigilance and to collect information on aDSM (active drug safety monitoring and management of adverse events-AEs).Aim of this prospective study was to evaluate the frequency and severity of AEs to anti-tuberculosis (TB) drugs in a cohort of consecutive TB patients treated with new (i.e. bedaquiline, delamanid) and repurposed (i.e. clofazimine, linezolid) drugs, based on the WHO aDSM project.AEs were collected prospectively after attribution to a specific drug together with demographic, bacteriological, radiological and clinical information at diagnosis and during therapy. This interim analysis included patients who completed or were still on treatment at time of data collection.Globally, 45 centres from 26 countries/regions reported 658 patients (68.7% males, 4.4% HIV co-infected) treated as follows: 87.7% with bedaquiline, 18.4% with delamanid (6.1% with both), 81.5% with linezolid and 32.4% with clofazimine. Overall 504 AE episodes were reported: 447 (447/504, 88.7%) were classified as minor (grade 1-2) and 57 (57/504, 11.3%) as serious (grade 3-5).The majority of the 57 serious AEs reported by 55 patients (51/57; 89.5%) ultimately resolved. Among patients reporting serious AEs some drugs held responsible were discontinued: bedaquiline in 0.35% (2/577), delamanid in 0.8% (1/121), linezolid in 1.9% (10/536) and clofazimine in 1.4% (3/213) of patients.Serious AEs were reported in 6.9% (9/131) of patients treated with amikacin, 0.4% (1/221) with ethionamide/prothionamide, 2.8% (15/536) with linezolid and 1.8% (8/498) with cycloserine/terizidone.The aDSM study provided valuable information but implementation needs scaling-up to support patient-centred care.

15.
Phys Chem Chem Phys ; 2019 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-31595279

RESUMO

The adenosine A1 receptor (A1R) is one of four adenosine receptors in humans, which are involved in the function of the cardiovascular, respiratory and central nervous systems. Experimental results indicate that A1R can form a homodimer and that the protomer-protomer interaction in the A1R dimer is related to certain pharmacological characteristics of A1R activation. In this work, we performed docking, metadynamics simulation, conventional molecular dynamics simulations, Gaussian-accelerated molecular dynamics simulations, potential of mean force calculations, dynamic cross-correlation motions analysis and community network analysis to study the binding mode of 5'-N-ethylcarboxamidoadenosine (NECA) to A1R and the effect of dimerization on the activation of A1R. Our results show that NECA binds to A1R in a similar mode to adenosine in the A1R crystal structure and NECA in the A2AR crystal structure. The A1R homodimer can be activated by one or two agonists with NECA occupying its orthosteric pockets in one (which we call the NECA-A1R system) or both protomers (which we call the dNECA-A1R system). In the NECA-A1R system, activation is predicated in the protomer without NECA bound. In the dNECA-A1R system, only one protomer achieves the active state. These findings suggest an asymmetrical activation mechanism of the homodimer and a negative cooperativity between the two protomers. We envision that our results may further facilitate the drug development of A1R.

16.
J Zhejiang Univ Sci B ; 20(11): 877-890, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31595724

RESUMO

Glycerol monolaurate (GML) has been widely used as an effective antibacterial emulsifier in the food industry. A total of 360 44-week-old Hy-Line brown laying hens were randomly distributed into four groups each with six replicates of 15 birds, and fed with corn-soybean-meal-based diets supplemented with 0, 0.15, 0.30, and 0.45 g/kg GML, respectively. Our results showed that 0.15, 0.30, and 0.45 g/kg GML treatments significantly decreased feed conversion ratios (FCRs) by 2.65%, 7.08%, and 3.54%, respectively, and significantly increased the laying rates and average egg weights. For egg quality, GML drastically increased albumen height and Haugh units, and enhanced yolk color. Notably, GML increased the concentrations of polyunsaturated and monounsaturated fatty acids and reduced the concentration of total saturated fatty acids in the yolk. The albumen composition was also significantly modified, with an increase of 1.02% in total protein content, and increased contents of His (4.55%) and Glu (2.02%) under the 0.30 g/kg GML treatment. Additionally, GML treatments had positive effects on the lipid metabolism of laying hens, including lowering the serum triglyceride and total cholesterol levels and reducing fat deposition in abdominal adipose tissue. Intestinal morphology was also improved by GML treatment, with increased villus length and villus height to crypt depth ratio. Our data demonstrated that GML supplementation of laying hens could have beneficial effects on both their productivity and physiological properties, which indicates the potential application of GML as a functional feed additive and gives us a new insight into this traditional food additive.

17.
Artigo em Inglês | MEDLINE | ID: mdl-31596040

RESUMO

A convenient and efficient domino aryne process was developed under transition metal-free conditions, featuring a group of tetra- and pentacyclic ring systems. This transformation was realized by a 1,2-benzdiyne-involved nucleophilic, Diels-Alder reaction cascade using styrene as diene moiety. Three new chemical bonds, namely one C-N and two C-C bonds, and two benzofused rings could be concomitantly constructed, which was made possible by distinct chemoselective control on both 1,2-aryne and 2,3-aryne stages. Moreover, in-depth studies were carried out on domino aryne precursors and diastereoselective control.

18.
Biomacromolecules ; 2019 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-31596574

RESUMO

Catheter-related infection is a great challenge to modern medicine, which causes significant economic burden and increases patient morbidity. Hence, there is a great requirement for functionalized surfaces with inherently antibacterial properties and biocompatibility that prevent bacterial colonization and attachment of blood cells. Herein, we developed a strategy for constructing polymer brushes with hierarchical architecture on polyurethane (PU) via surface-initiated atom-transfer radical polymerization (SI-ATRP). Surface-functionalized PU (PU-DMH) was readily prepared, which comprised of poly(3-[dimethyl-[2-(2-methylprop-2-enoyloxy)ethyl]azaniumyl]propane-1-sulfonate) (PDMAPS) brushes as the lower layer and antimicrobial peptide-conjugated poly (methacrylic acid) (PMAA) brushes as the upper layer. The PU-DMH surface showed excellent bactericidal property against both Gram-positive and Gram-negative bacteria, and could prevent accumulation of bacterial debris on surfaces. Simultaneously, the PU-DMH samples possessed good hemocompatibility and low cytotoxicity. Furthermore, the integrated antifouling and bactericidal properties of PU-DMH under hydrodynamic conditions were confirmed by an in vitro circulating model. The functionalized surface possessed persistent antifouling and bactericidal performances both under static and hydrodynamic conditions. The microbiological and histological results of animal experiments also verified the in vivo anti-infection performance. The present work might find promising clinical applications for preventing catheter-related infection.

19.
J Cell Mol Med ; 2019 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-31568645

RESUMO

Diabetic nephropathy (DN) remains one of the severe complications associated with diabetes mellitus. It is worthwhile to uncover the underlying mechanisms of clinical benefits of human urine-derived stem cells (hUSCs) in the treatment of DN. At present, the clinical benefits associated with hUSCs in the treatment of DN remains unclear. Hence, our study aims to investigate protective effect of hUSC exosome along with microRNA-16-5p (miR-16-5p) on podocytes in DN via vascular endothelial growth factor A (VEGFA). Initially, miR-16-5p was predicated to target VEGFA based on data retrieved from several bioinformatics databases. Notably, dual-luciferase report gene assay provided further verification confirming the prediction. Moreover, our results demonstrated that high glucose (HG) stimulation could inhibit miR-16-5p and promote VEGFA in human podocytes (HPDCs). miR-16-5p in hUSCs was transferred through the exosome pathway to HG-treated HPDCs. The viability and apoptosis rate of podocytes after HG treatment together with expression of the related factors were subsequently determined. The results indicated that miR-16-5p secreted by hUSCs could improve podocyte injury induced by HG. In addition, VEGA silencing could also ameliorate HG-induced podocyte injury. Finally, hUSC exosomes containing overexpressed miR-16-5p were injected into diabetic rats via tail vein, followed by qualification of miR-16-5p and observation on the changes of podocytes, which revealed that overexpressed miR-16-5p in hUSCs conferred protective effects on HPDCs in diabetic rats. Taken together, the present study revealed that overexpressed miR-16-5p in hUSC exosomes could protect HPDCs induced by HG and suppress VEGFA expression and podocytic apoptosis, providing fresh insights for novel treatment of DN.

20.
Int J Gynecol Cancer ; 29(8): 1280-1284, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31570543

RESUMO

INTRODUCTION: The solute carrier family 12 member 5 (SLC12A5) gene is playing a putative oncogenic role in colorectal carcinoma. However, the status of SLC12A5 amplification and expression in ovarian carcinoma and its potential clinical and/or prognostic significance has not yet been investigated. METHODS: In the present study, semi-quantitative staining and fluorescence in situ hybridization were used to investigate SLC12A5 protein expression and gene amplification levels. Samples were obtained from archival, formalin-fixed, paraffin-embedded pathological specimens consisting of 30 normal ovaries, 30 ovarian cystadenomas, 30 borderline ovarian tumors, and 147 invasive ovarian carcinomas. SLC12A5 immunohistochemical staining results, pathological parameters, and patient prognosis were then evaluated using various statistical models. Patient survival rate was also assessed using receiver-operator curve analysis. RESULTS: Our results revealed no SLC12A5 protein overexpression in normal ovaries. However, 7% of cystadenomas had SLC12A5 protein overexpression along with 17% of borderline tumors and 37% of ovarian carcinomas (P<0.01). Amplification of SLC12A5 was detected in 10.3% of ovarian carcinomas. Further correlational analyses showed that SLC12A5 protein overexpression in ovarian carcinomas was significantly associated with ascending histological grade, pT/pN/pM status, as well as FIGO stage (P<0.05). A subsequent univariate survival analysis of our ovarian carcinoma cohorts resulted in a significant association between SLC12A5 protein overexpression and decreased patient survival (44.3 and 85.9 months for high and low SLC12A5 protein expression, respectively; P<0.001). Importantly, additional multivariate analysis revealed that SLC12A5 protein expression was a significant, independent prognostic factor for overall survival in ovarian carcinoma patients (P=0.003). CONCLUSIONS: Collectively, these findings support the conclusion that SLC12A5 protein overexpression could indicate an invasive and/or aggressive phenotype of ovarian carcinoma. Future work will need to investigate whether SLC12A5 protein can serve as an independent prognostic molecular marker in patients with ovarian carcinoma.

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