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Objective:To investigate the effects of tamoxifen on high glucose-induced epithelial-to-mesenchymal transition of rat peritoneal mesothelial cells and the underlying mechanism.Methods:The peritoneal mesothelial cells of normal male SD rats were selected between January 2015 and June 2016 and then cultured and divided into blank control, high-glucose stimulation and drug intervention groups. High-glucose stimulation group: primary cultured rat peritoneal mesothelial cells (RPMCs) were treated with 60 mmol/L high-concentration glucose to induce epithelial-to-mesenchymal transition. Drug intervention group: (1) RPMCs were treated with 60 mmol/L high-concentration glucose and different concentrations (0.5 μmol/L, 2 μmol/L) of tamoxifen. After 72 hours of stimulation, protein was extracted. (2) RPMCs were treated with 60 mmol/L high-concentration glucose and 2 μmol/L tamoxifen with or without 2 μmol/L ER-α antagonist for 1 hour to extract protein and for 6 hours to extract RNA. (3) RPMCs were treated with high-concentration glucose and 2 μmol/L tamoxifen with or without 1 μmol/L 1 μM proteasome inhibitor for 1 hour to extract protein. Western blot analysis was performed to analyze change in E-cadherin, α-SMA, Smad2, p-Smad2, Smad3, p-Smad3 and Smad4 protein. Real-time fluorescence quantitative PCR was performed to detect the change in mRNA expression of Smad2, Smad3, connective tissue growth factor and plasminogen activator inhibitor 1.Results:Tamoxifen attenuated epithelial-to-mesenchymal transition on RPMCs induced by high-level glucose, showing increased expression of epithelial cell marker E-cadherin and decreased expression of α-SMA in a concentration-dependent manner ( tE-cadherin = 2.31, tα-SMA =-2.53, both P < 0.05).TGF-β1/R-Smad signal pathway was activated by high-concentration glucose. Phosphorylation of Smad2/3 and mRNA expression of CTGF and PAI-1 were increased. Tamoxifen remarkably reduced protein and mRNA level of above mentioned protein and related target genes ( tp-Smad2 = -3.38, tCTGF = -3.81, P < 0.05), which could be blocked by ER-α antagonist. Finally, proteasome inhibitor could weaken the inhibitory effects of tamoxifen on p-Smad2/3 and increase p-Smad2/3 protein level ( tp-Smad2 = 3.94, P < 0.05). Conclusion:Tamoxifen activates ER-α on RPMCs, weakens the activation of TGF-β1/R-Smad signal pathway through decreasing p-Smad2 protein level, and effectively inhibits the progression of high-concentration glucose-induced epithelial-to-mesenchymal transition possibly through degrading p-Smad2 protein through proteasome. The role of tamoxifen in epithelial-to-mesenchymal transition may provide a possible guide for research, prevention and treatment of peritoneal fibrosis.
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Objective@# To develop a magnetic solid-phase extraction combined with gas chromatography-tandem mass spectrometry (GC-MS/MS) based on pyrrole-modified magnetic nanoparticles to determine 16 types of phthalic acid esters (PAEs) in commercial liquors.@*Methods @#Fe3O4 magnetic nuclei were prepared by hydrothermal synthesis, and the polypyrrole-modified magnetic nanomaterials were prepared with the chemical oxidation method. Magnetic solid-phase extraction of beer, grape wine, rice wine and Chinese spirits was performed at 10% alcohol by volume, extraction duration of 20 min and ethyl acetate elution of 10 min, followed by addition of 1 g NaCl for reduction of emulsification effect. The 16 types of PAEs were determined using GC-MS/MS with DB-5MS capillary column (30 m×250 μm, 0.25 μm) under the mode of electron impact ionization (EI) and dynamic multiple reaction monitoring (dMRM), with quantitative analysis using the external standard method. The standard curve, detection limit, spike recovery rate and precision of GC-MS/MS for determination of 16 types of PAEs were evaluated.@*Results@#Pyrrole was successfully embedded onto the surface of magnetic nanoparticles in the form of polymer, and the magnetic nanoparticles modified by polypyrrole were well characterized, showing unapparent matrix and emulsification effects. There was a good linear relationship for the 16 types of PAEs at 50 to 5 000 ng/mL (r=0.999 5-0.999 9), and the spike recovery rate of 16 types of PAEs ranged from 71.61% to 110.50% at 100, 500 and 1 000 μg/kg, with relative standard deviations of 3.78% to 7.41%, detection limits of 0.02 to 1.47 μg/kg. PAEs were detected in 20 out of 50 liquor samples, with 30.00%, 60.00%, 40.00% and 70.00% detection rates in beer, grape wine, rice wine and Chinese spirits, respectively. @*Conclusions@#This method is sensitive to determine 16 types of PAEs in liquor samples, with unapparent matrix and emulsification effects, and the polypyrrole-modified magnetic composite nanoparticles present high adsorption of PAEs in liquor samples, which is feasible for monitoring of PAEs in multiple types of liquor samples.
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Objective:To understand the iodine nutrition status of pregnant women in Huzhou City, and to provide a basis for timely targeted prevention and control of the iodine deficiency disorders.Methods:In 2017, five districts (counties) in Wuxing, Nanxun, Deqing, Changxing, and Anji were used as monitoring sites in Huzhou City, and each monitoring site was divided into five areas according to the east, west, south, north, and middle locations. One township (street) in each location was sampled to collect drinking water samples from residents, and the water iodine content was tested; 21 pregnant women (7 each in the early, middle, and late pregnancy periods) were sampled in each township (street), and edible salt samples were collected at home for testing salt iodine and urine samples were collected for testing urinary iodine content.Results:A total of 332 drinking water samples were collected from Huzhou residents, water iodine median was 2.2 μg/L, which belonged to iodine-deficient regions; and there was statistically significant difference in the water iodine median between different regions ( H = 161.0, P < 0.05). And 525 edible salt samples were detected from pregnant women, the salt iodine median was 23.4 mg/kg, the salt iodine coverage was 97.1% (510/525), and the qualified iodized salt consumption rate was 93.0% (488/525). There was a statistically significant difference in salt iodine levels between different regions ( H = 67.7, P < 0.05). Meanwhile, the urinary iodine median of 525 urine samples of pregnant women was 123.1 μg/L, which was at the level of iodine deficiency. From the regional distribution, the urinary iodine median in Deqing County was 154.0 μg/L, which was the highest and at an iodine appropriate level; the other regions were Wuxing District, Nanxun District, Anji County, and Changxing County in order, all of them were at the iodine deficiency level; the urinary iodine median of pregnant women between different regions was statistically significantly different ( H = 14.1, P < 0.05). From the pregnancy periods' distribution, the urinary iodine median in middle pregnancy period was the highest at 172.8 μg/L; there was statistically significant difference between different pregnancy periods ( H = 7.5, P < 0.05). Conclusions:The Huzhou City belongs to the environmental iodine deficiency area. Except for Deqing County, the iodine nutrition status of pregnant women in other districts (counties) is at an iodine deficiency level. Urinary iodine level monitoring and health education of pregnant women should be strengthened to improve iodine nutrition status.
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Objective:To elucidate the effects of andrographolide (AG) against acute pancreatitis (AP)-associated kidney injury and explore the potential mechanism.Methods:Thirty-two C57BL/6 mice were divided into control group, AP group, AG treatment group and AG control group. In AP group, AP model was established by intraperitoneal injection of L-arginine and LPS. AG treatment group was intravenously injected with AG 8 h before the model establishment. The control group was intraperitoneally injected with an equal volume of normal saline. The AG control group was intravenously injected with AG 8 h before the intraperitoneal administration of normal saline. Venous blood of inner canthus in mice was collected, and the levels of amylase, creatinine (Scr) and urea nitrogen (BUN) in serum were detected. Pancreatic and double kidney tissues were also collected. Pancreatic myeloperoxidase (MPO) activity was detected by colorimetry. Histopathological changes of pancreas and kidneys were observed under light microscope and pathological score was evaluated. Ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry was used to analyze metabonomics in renal tissue.Results:Compared to the control group, the levels of amylase, Scr and BUN in serum and pancreatic MPO activity in the AP group were obviously increased [(13.78±6.01)U/L to (0.23±0.04)U/L, (79.81±24.03)μmol/L to (9.02±2.87)μmol/L, (34.76±14.53)mmol/L to (8.52±2.55)mmol/L, (16.55±4.23)U/g pro to (2.32±0.55)U/g pro]; histopathologic scores of pancreas and kidney in the AP group were also increased (2.70±0.26 to 0.20±0.12, 3.00±0.35 to 0.30±0.12), and the differences were statistically significant (all P<0.05). The levels of amylase, Scr and BUN in serum and pancreatic MPO activity in the AG treatment group were decreased than the AP group [(8.26±3.87)U/L to (13.78±6.01)U/L, (55.42±17.25)μmol/L to (79.81±24.03)μmol/L, (20.66±10.30)mmol/L to (34.76±14.53)mmol/L, (11.51±3.29)U/g pro to (16.55±4.23)U/g pro]; histopathologic scores of pancreas and kidney were also decreased in the AG treatment group than those in the AP group (1.40±0.19 to 2.70±0.26、1.70±0.26 to 3.00±0.35), and the differences were statistically significant (all P<0.05). Metabonomics analysis detected a total of 31 metabolites. Most of metabolites were involved in amino acid and fatty acid metabolism, and a few of them were also involved in glucose, nucleotide, vitamin and bile acid metabolism. Conclusions:AG exerts protective effects on AP-associated kidney injury by altering the levels of multiple metabolites.
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Objective To investigate the potential mechanism of Rhizoma Coptidis extracts (RCE) against sepsis associated with acute kidney injury. Methods C57BL/6 mice were divided into sham group, model group and RCE treatment group. The levels of Scr and BUN were measured by test kits. Gas chromatography-mass spectrometry was used to analyze metabolic changes in kidneys. Results The levels of Scr and BUN were increased in the model group than sham, which were reversed by RCE. 16 metabolites related to the progress of sepsis associated with acute kidney injury were detected, which were involved in amino acid metabolism and carbohydrate metabolism. Among these metabolites, the level of 8 metabolites can be reversed with RCE treatment. Conclusion RCE might exert therapeutic effects in sepsis associated with acute kidney injury by altering multiple metabolic pathways.
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Objective To investigate the diagnostic and prognostic value of neutrophil-to-lymphocyte rate ( NLR) in acute pancreatitis related acute kidney injury patients .Methods Peripheral blood specimens and clinical information of 98 acute pancreatitis patients in Wenzhou Center Hospital were collected .The WBC, neutrophils , lymphocytes were detected and NLR were calculated when they were admitted .The patients were divided into two groups by their NLR and whether they were with AKI respectively .Besides, ALT, Hct, TC, TG, blood calcium concentration , serum creatine and urea nitrogen , C reactive protein were detected and patients′APACHEⅡ score were also recorded to analyze the difference between the two groups .Results There is no significance in the age , BMI, Hct, TG, TC, ALT and blood calcium between AP patients with and without AKI.The blood creatinine, BUN, CRP, APACHEⅡscore were (395 ±122)μmol/L, (28.2 ±5.2) mmol/L, (34.0 ±8.2)mg/L, (11.5 ±3.8) score, respectively in AP patients with AKI, and which were (79 ±17 )μmol/L, ( 7.3 ±2.0 ) mmol/L, ( 14.8 ±2.9 ) mg/L, ( 6.9 ±2.4 ) score, respectively in AP patients without AKI.The blood ALT, blood creatinine, BUN, CRP, APACHEⅡscore were (257 ±76)U/L, (159 ±62)μmol/L, (20.5 ±6.6)mmol/L, (24.8 ±5.5)mg/L and (12.4 ±4.6) score in the patients with higher NLR respectively , and which were ( 165 ±30 ) U/L, ( 98 ±23 )μmol/L, ( 14.3 ±5.2 ) mmol/L, (19.5 ±3.0)mg/L and (5.4 ±2.1) score in the patients with lower NLR respectively .NLR was 4.97 ±0.19 in AP patients with AKI, and was 9.62 ±0.81 in AP patients without AKI.The difference between the two groups was significant(P=0.0001).The area under ROC curve of diagnosing AP by NLR was 0.895 (95%CI 5.75).the sensitivity was 89.5%and the specificity was 77.2% when using 5.75 as the cut-off value to diagnose AP related AKI with NLR .Conclusions NLR can be a potential predictive index of the severity and relate to renal function in acute pancreatitis related acute kidney injury patients .
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<p><b>OBJECTIVE</b>To develop a method for determination of mandelic acid (MA) and phenylglyoxylic acid (PGA) in urine by reagent-free ion chromatography.</p><p><b>METHODS</b>Ion chromatography was performed on an AS19 column with a gradient elution solution containing 10-35 mmoL/L KOH at a flow rate of 1.00 ml/min, and MA and PGA were detected at ultraviolet wavelengths of 225 nm and 254 nm, respectively. The samples were diluted 10 times with purified water, then purified on a silver column to remove high concentrations of chloride ion, and injected after being filtered through a 0.2-µm m filter membrane.</p><p><b>RESULTS</b>The recoveries of standard addition of MA and PGA were 96.5% and 99.3%, respectively, with both relative standard deviations less than 5.0%. Good linear relationships were noted in the range of 1.0-100.0 mg/L for both MA and PGA (r >0.9995). The detection limits of MA and PGA were 0.02 mg/L and 0.05 mg/L, respectively; the minimum detectable concentrations of MA and PGA were 0.2 mg/L and 0.5 mg/L (when the sampling amount was 5.0 ml and diluted to 50.0 ml with water, and the injection volume was 300 µL).</p><p><b>CONCLUSIONS</b>This method is fast, convenient, and highly sensitive and selective. It can be used for the analysis of MA and PGA in the urine of styrene-exposed workers.</p>