Pueraria isoflavones inhibit XOD and GLUT9 to decrease uric acid production and promote uric acid excretion, respectively

Objective: To analyze the possible mechanism of Pueraria isoflavones inhibiting XOD and GLUT9 to reduce uric acid production and promote uric acid excretion. Methods: August 2021-April 2022, a total of forty SPF male Kunming mice were divided into the healthy group (carboxymethylcellulose sodium at a dose of 250 mg/kg), the model group (HUA mice were given carboxymethylcellulose sodium at a dose of 250 mg/kg), the low group (HUA mice were given pueraria isoflavone at a dose of 125 mg/kg), HUA mice were given pueraria isoflavones at a dose of 250 mg/kg once d frequency)and the high group (HUA mice were given pueraria isoflavones at a dose of 500 mg/kg once d frequency) dosage groups, with 8 mice in each group. The contents of uric acid (SUA), urea nitrogen (BUN) and creatinine (SCr) in serum and urine of each group were compared before and after intervention (30 d). Statistical differences of xanthine oxidase (XOD) and human glucose transporter 9(GLUT9), cy- clooxygenase- 2(COX-2), tumor necrosis factor (TNF-α) and interleukin-1 (IL-1β) contents in renal tissues of each group after intervention (30 d) were compared. Results: After intervention, kidney inflammatory factors (COX-2, TNF-α and IL-1β) in the model group were compared. Blood and urine indexes (SUA, BUN, SCr);The contents of XOD and GLUT9 were higher than those of healthy group(P<0.05). Renal inflammatory cytokines (COX-2, TNF-α and IL-1β) in low, medium and high dose groups;Blood and urine indexes (SUA, BUN, SCr);The contents of XOD and GLUT9 were lower than those of model group, and there were low > medium > high dose groups, the comparison between the two groups had statistical significance(P< 0.05). After intervention, the contents of 3 indicators in blood or urine(COX-2, TNF-α and IL-1β) all decreased compared with before intervention, and the differences in intra-group comparison were statistically significant (P<0.05). Conclusion: Pueraria isoflavones can treat HUA mice by inhibiting the expression of XOD and GLUT9, and then play a role in reducing uric acid pro- duction and promoting uric acid excretion, as well as alleviating the degree of disease inflammation.

Development and Validation of UHPLC-MS/MS Method for Quantifying of Agarotriose:An Application for Pharmacokinetic,Tissue Distribution,and Excretion Studies in Rats

A sensitive,rapid,and robust ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)method was established for the first time to quantify agarotriose(A3)in rat plasma,tissues,urine,and feces.A3 and stachyose(internal standard)were separated by a BEH amide column at 65℃under the mobile phase of 10 mmol L^(-1)ammonium ace-tate-acetonitrile(42:58,v/v)with 350µLmin-1.The acquisition of transitions was carried out in multiple reaction monitoring(MRM)pattern operating with positive ionization at m/z 509.16>329.15 for A3 and m/z 689.15>527.11 for stachyose.The linearity ranges of A3 were 10 to 5000nmolL^(-1)for plasma,20 to 10000nmolL^(-1)for tissues,and 40 to 20000nmolL^(-1)for urine and feces.The accuracy and precision ranged from 90.9%to 111.6%and 0.7%to 10.1%,respectively.The stability was between 86.1%and 102.5%.The extraction recovery was consistent and reproducible.The matrix effect ranged from 1.5%to 11.4%.The pharmacokinetic,tissue dis-tribution,and excretion studies were successfully conducted with the validated method.Results showed that A3 could be absorbed by rats,and the absolute bioavailability was 6.7%.Furthermore,it was rapidly distributed in rat tissues and mainly eliminated via feces excretion(67.0%)after oral administration.For intravenous bolus,85.5%was recovered,and renal excretion was the primary path-way(77.6%)for cumulative recovery.

Dietary nutrient status modulates nutrient regeneration in the marine ciliate Euplotes vannus

Marine ciliates play important roles not only in linking the microbial loop to the classic pelagic and benthic food chains but also in regenerating nutrients,yet how dietary nutrient imbalance impacts their nutrient regeneration has not been thoroughly addressed.The growth and physiological responses of Euplotes vannus to low dietary nitrogen(LN)and low dietary phosphorus(LP)conditions were studied,with the bacterium Pseudomonas putida as prey.Feeding on LN prey reduced the growth rate of E.vannus.Dietary nutrient limitation changed the types and quantities of nutrient recycling.Feeding on LP prey enhanced dissolved organic carbon excretion but reduced orthophosphate excretion,whereas feeding on LN prey generally resulted in decreases in the excretion rate in all N forms(ammonium,urea,and nitrate).In addition,the proportion of ammonium in regenerated N increased significantly under the LN condition.These findings indicate that a nutrient-imbalanced diet triggers E.vannus to retain limited macronutrients and promotes the recycling of excessive macronutrients,which may potentially form positive and negative feedback to ambient N and P limitations,respectively.

Albumin-bound kynurenic acid is an appropriate endogenous biomarker for assessment of the renal tubular OATs-MRP4 channel

Renal tubular secretion mediated by organic anion transporters(OATs)and the multidrug resistanceassociated protein 4(MRP4)is an important means of drug and toxin excretion.Unfortunately,there are no biomarkers to evaluate their function.The aim of this study was to identify and characterize an endogenous biomarker of the renal tubular OATs-MRP4 channel.Twenty-six uremic toxins were selected as candidate compounds,of which kynurenic acid was identified as a potential biomarker by assessing the protein-binding ratio and the uptake in OAT1-,OAT3-,and MRP4-overexpressing cell lines.OAT1/3 and MRP4 mediated the transcellular vectorial transport of kynurenic acid in vitro.Serum kynurenic acid concentration was dramatically increased in rats treated with a rat OAT1/3(rOAT1/3)inhibitor and in rOAT1/3 double knockout(rOAT1/3^(-/-))rats,and the renal concentrations were markedly elevated by the rat MRP4(rMRP4)inhibitor.Kynurenic acid was not filtered at the glomerulus(99%of albumin binding),and was specifically secreted in renal tubules through the OAT1/3-MRP4 channel with an appropriate affinity(Km)(496.7 mM and 382.2 mM for OAT1 and OAT3,respectively)and renal clearance half-life(t1/2)in vivo(3.7±0.7 h).There is a strong correlation in area under the plasma drug concentration-time curve(AUC0et)between cefmetazole and kynurenic acid,but not with creatinine,after inhibition of rOATs.In addition,the phase of increased kynurenic acid level is earlier than that of creatinine in acute kidney injury process.These results suggest that albumin-bound kynurenic acid is an appropriate endogenous biomarker for adjusting the dosage of drugs secreted by this channel or predicting kidney injury.

Analysis of Clinical Characteristics of Patients with Different Types of Primary Hyperuricemia

Objective:This paper aims to study the clinical characteristics of patients with different types of primary hyperuricemia(HUA).Methods:Using a retrospective research method,200 patients with primary HUA in the hospital from June 2020 to January 2023 were selected as the research objects.Patients were grouped according to the detection results of 24-hour urinary uric acid excretion(UUE)and fractional excretion of uric acid(FEUA)(renal insufficiency type,renal overload type,mixed type,and other types).The general information of patients in the four groups(gender,age,body mass index,living habits,etc.),underlying diseases(hypertension,diabetes),blood test results[uric acid(UA),creatinine(Cre)],urine test results(24-hour urine UA,24-hour urine Cre)were summarized and the differences between the groups were analyzed.Results:The 200 cases of HUA patients were divided into 54.00%with renal insufficiency type,38.50%with mixed type,6.00%with renal overload type,and 1.50%with other types.The age of patients with mixed HUA was younger than that of patients with other types,renal overload type,and renal insufficiency type,and the difference was statistically significant(P<0.05).The UA level of patients with other types of HUA was lower than that of patients with mixed type HUA,and there was statistical significance(P<0.05).The Cre level of patients with mixed type HUA was lower than that of patients with renal insufficiency type and renal overload type,and the difference was statistically significant(P<0.05).The 24-hour urinary UA level in patients with renal insufficiency type HUA was lower than that in patients with renal overload type and mixed type HUA,and the difference was statistically significant(P<0.05).The 24-hour urinary Cre level of patients with other types of HUA was lower than that of patients with renal overload type and mixed HUA,the difference was statistically significant(P<0.05).The estimated glomerular filtration rate(eGFR)level of patients with other types of HUA was lower than that of patients with mixed type HUA,and the difference was statistically significant(P<0.05).There was no significant difference in the proportions of hypertension,diabetes,coronary heart disease,and urinary calculi among patients with renal insufficiency type,renal overload type,mixed type,and other types of HUA(P>0.05).Conclusion:The primary HUA patients are mainly of renal insufficiency type,followed by mixed type.There are significant differences in the clinical characteristics of patients with different types of HUA.Among them,patients with other types of HUA are the oldest and have the lowest uric acid levels.Patients with mixed HUA had the best renal function but the highest 24-hour urine creatinine level.This study can be used as a basis for rational selection of urate-lowering drugs for different HUA patients.

Biomarkers in diabetic nephropathy: Present and future

Diabetic nephropathy(DN) is the leading cause of end stage renal disease in the Western world. Microalbuminuria(MA) is the earliest and most commonly used clinical index of DN and is independently associated with cardiovascular risk in diabetic patients. Although MA remains an essential tool for risk stratification and monitoring disease progression in DN, a number of factors have called into question its predictive power. Originally thought to be predictive of future overt DN in 80% of patients, we now know that only around 30% of microalbuminuric patients progress to overt nephropathy after 10 years of follow up. In addition, advanced structural alterations in the glomerular basement membrane may already have occurred by the time MA is clinically detectable.Evidence in recent years suggests that a significant proportion of patients with MA can revert to normoalbuminuria and the concept of nonalbuminuric DN is well-documented, reflecting the fact that patients with diabetes can demonstrate a reduction in glomerular filtration rate without progressing from normo-to MA. There is an unmet clinical need to identify biomarkers with potential for earlier diagnosis and risk stratification in DN and recent developments inthis field will be the focus of this review article.

Linking uric acid metabolism to diabetic complications

Hyperuricemia have been thought to be caused by the ingestion of large amounts of purines, and prevention or treatment of hyperuricemia has intended to prevent gout. Xanthine dehydrogenase/xanthine oxidase(XDH/XO) is rate-limiting enzyme of uric acid generation, and allopurinol was developed as a uric acid(UA) generation inhibitor in the 1950 s and has been routinely used for gout prevention since then. Serum UA levels are an important risk factor of disease progression for various diseases, including those related to lifestyle. Recently, other UA generation inhibitors such as febuxostat and topiroxostat were launched. The emergence of these novel medications has promoted new research in the field. Lifestyle-related diseases, such as metabolic syndrome or type 2 diabetes mellitus, often have a common pathological foundation. As such, hyperuricemia is often present among these patients. Many in vitro and animal studies have implicated inflammation and oxidative stress in UA metabolism and vascular injury because XDH/XO act as one of the major source of reactive oxygen species Many studies on UA levels and associated diseases implicate involvement of UA generation in disease onset and/or progression. Interventional studies for UA generation, not UA excretion revealed XDH/XO can be the therapeutic target forvascular injury and renal dysfunction. In this review, the relationship between UA metabolism and diabetic complications is highlighted.

Effect of different dietary protein levels and amino acids supplementation patterns on growth performance, carcass characteristics and nitrogen excretion in growing-finishing pigs

Background:This study was conducted to determine the effects of different dietary protein levels and amino acids supplementation patterns in low protein diets on the growth performance,carcass characteristics and nitrogen excretion in growing-finishing pigs.Forty-two barrows(25.00±0.39 kg)were randomly assigned to 7 diets.Diet 1:the high crude protein diet with balanced for 10 essential amino acids(EAAs).Diet 2:the medium crude protein diet with 2%(approx)decreased protein level of Diet 1 and balanced 10 EAAs.Diet 3:the low crude protein diet with 4%decreased protein level of Diet 1 and balanced 10 EAAs.The protein levels of Diet 4,5,6 and 7 were the same as that of Diet 3.Diet 4 was only balanced for lysine(Lys),methionine(Met),threonine(Thr)and tryptophan(Trp);Diet 5 and 6 were further supplemented with extra isoleucine(Ile)or valine(Val),respectively;Diet 7 was further supplemented with extra Ile+Val.Results:Over the 112 days trial,the reduction of dietary protein by 2%or 4%with balanced10 EAAs significantly decreased nitrogen excretion(P<0.05),but had no effects on growth performance and carcass characteristics(P>0.05).In low protein diet,Val supplementation significantly increased body weight gain at 25–50 kg phase(P<0.05),while Ile supplementation at 75–100 kg phase and 100–125 kg phase significantly reduced the ratio of feed to gain(P<0.05).No effect of different dietary protein levels and amino acids supplementation patterns in low protein diets on carcass characteristics was observed(P>0.05).The total N excretion of pigs supplemented with only Lys,Met,Thr and Trp was numerically higher than that of pigs fed with extra Ile,or Val,or Ile+Val diets.Conclusion:In low protein diet,Val is more required than Ile in the early growing phage(25–50 kg),while Ile becomes more required in the late growing and finishing phage(75–125 kg).

Pharmacologic inducers of the uric acid exporter ABCG2 as potential drugs for treatment of gouty arthritis

Uric acid is the end product of purine catabolism and its plasma levels are maintained below its maximum solubility in water(6–7 mg/dl).The plasma levels are tightly regulated as the balance between the rate of production and the rate of excretion,the latter occurring in urine(kidney),bile(liver)and feces(intestinal tract).Reabsorption in kidney is also an important component of this process.Both excretion and reabsorption are mediated by specific transporters.Disruption of the balance between production and excretion leads to hyperuricemia,which increases the risk of uric acid crystallization as monosodium urate with subsequent deposition of the crystals in joints causing gouty arthritis.Loss-of-function mutations in the transporters that mediate uric acid excretion are associated with gout.The ATP-Binding Cassette exporter ABCG2 is important in uric acid excretion at all three sites:kidney(urine),liver(bile),and intestine(feces).Mutations in this transporter cause gout and these mutations occur at significant prevalence in general population.However,mutations that are most prevalent result only in partial loss of transport function.Therefore,if the expression of these partially defective transporters could be induced,the increased number of the transporter molecules would compensate for the mutation-associated decrease in transport function and hence increase uric acid excretion.As such,pharmacologic agents with ability to induce the expression of ABCG2 represent potentially a novel class of drugs for treatment of gouty arthritis.

Nutritional strategies for reducing nitrogen, phosphorus and trace mineral excretions of livestock and poultry

Animal agriculture contributes to environmental pollutions through the surplus nitrogen（N）, phosphorus（P）, and trace minerals that animals excrete. Animal nutritionists have sought alternatives to formulate more efficient diets and reduce production costs and environmental concerns. In general, element excretions may be reduced by avoiding the overfeeding of specific elements or using nutritional approaches to improve element utilizations by the animals. Several nutritional strategies are possible for minimizing N, P, and trace mineral excretions： 1） to accurately meet dietary N, P and trace mineral requirements of animals, which includes reducing the dietary crude protein contents with synthetic amino acids or feeding high rumenally undegraded protein, minimizing the adequate levels of dietary P and trace elements, adopting phase or group-feeding program, and considering the bioavailable trace mineral contents in the feed ingredients; 2） to improve the bioavailabilities of dietary N, P, and trace elements for animals by using some additives（enzymes, growth promoters, probiotics, prebiotics, vitamin D isomers, and organic acids）; 3） to use highly available P sources or organic trace elements. In the future, nutrient strategies must be integrated into total production systems so that animal production systems are environmentally safe as well as economically viable.

The Effect of Dietary Supplementation with Phytase Transgenic Corn on Growth Performance,Phosphorus Utilization and Excretion in Growing Pigs (Sus scrofa)

Two experiments were conducted to investigate the effect of dietary supplementation with phytase transgenic corn （PTC） on growth performance,phosphorus （P） utilization and excretion in growing pigs.In Exp.1,180 pigs （Large White × Landrace,BW=37.7 kg） were randomly allotted to 4 treatments with 5 replicates of 9 pigs each in order to evaluate the effect of PTC supplementation in low-P diets on growth performance.Four corn soybean meal-based diets consisted of a positive control （PC） diet,a diet containing 500 units （U） of exogenous phytase kg-1 （EP） on the basis of low-P （inorganic P reduced by 0.05% from PC diet） and the low-P＋500 （PTC1） or 750 （PTC2） phytase U of PTC kg-1.In Exp.2,20 barrows （Large White×Landrace,BW=31 kg,4 treatments with 5 replicates of 1 pig each） were randomly selected to evaluate the effect of PTC in low-P diets on serum parameters and nutrient utilization.Diets in Exp.2 were similar to those in Exp.1 except that the EP group was replaced by a low-P diet without exogenous phytase supplementation as a negative control （NC） group.The results from Exp.1 showed that the average daily gain （ADG） in the PTC2 group was significantly higher （P〈0.05） than that in the EP group over all periods.On the other hand,the feed：gain （F：G） ratio of the EP group was significantly higher （P〈0.05） than that of the PTC2 group during 1-21 and 1-42 d,respectively.There were no differences in average daily feed intake （ADFI） among all treatments （P〉0.05）.The results from Exp.2 showed that the concentration of serum Ca in the NC group was the highest （P〈0.05）,while the concentration of serum P in the PTC2 group was the highest （P〈0.05） among all treatments.There was a significant decrease （P〈0.05） in the P apparent digestibility of the NC group compared with the other groups,and that of PTC2 group was the best.Furthermore,fecal P excretion was reduced （P〈0.05） from 1.80 g d-1 in the PC group to 1.28 g d-1 in the PTC2 group.In conclusion,dietary supplementation with PTC could reduce the application of inorganic P,decrease fecal P excretion,and improve the growth performance of growing pigs.

Reverse cholesterol transport: From classical view to new insights

Cholesterol is of vital importance for the human body. It is a constituent for most biological membranes, it is needed for the formation of bile salts, and it is the pre- cursor for steroid hormones and vitamin D. However, the presence of excess cholesterol in cells, and in particular in macrophages in the arterial vessel wall, might be harmful. The accumulation of cholesterol in arteries can lead to atherosclerosis, and in turn, to other cardiovascular diseases. The route that is primarily thought to be responsible for the disposal of cholesterol is called reverse cholesterol transport (RCT). Therefore, RCT is seen as an interesting target for the development of drugs aimed at the prevention of atherosclerosis. Research on RCT has taken off in recent years. In this review, the classical concepts about RCT are discussed, together with new insights about this topic.

Association of urinary albumin excretion with central foveal thickness and intravitreal conbercept treatment frequency in patients with diabetic macular edema

AIM: To investigate the effect of albuminuria on diabetic macular edema(DME) and the possible association between baseline urinary albumin excretion(UAE) and intravitreal conbercept(IVC) treatment frequency in DME patients. METHODS: In this hospital-based retrospective study, a total of 350 in-patients with type 2 diabetes mellitus were recruited and their clinical records were reviewed. Thereafter, 52 patients identified with severe non-proliferative diabetic retinopathy(NPDR) combined with albuminuria were divided into the microalbuminuria(UAE 30-300 mg/24 h) and macroalbuminuria(UAE>300 mg/24 h) groups, which were compared and analyzed by both independent sample t-test and Chi-square test. Correlations between the systemic variables and the central foveal thickness(CFT) were evaluated using Spearman’s correlation and linear regression analyses. Of the 52 patients with centerinvolved DME, 43 received an initial combined injection of conbercept(0.5 mg/0.05 mL) and triamcinolone acetonide(1 mg/0.05 mL), followed by an IVC injection, as needed. The relationship between baseline UAE and number of IVCinjections during the first year of treatment was analyzed using Spearman’s partial correlation.RESULTS: Of 350 patients, a higher incidence of DME was observed in severe non-proliferative retinopathy(NPDR) patients than that observed in other groups. By dividing the 52 patients with severe NPDR into the micro-and macro-albuminuria subgroups, significant differences in CFT, systolic blood pressure, total cholesterol and serum creatinine levels, and UAE were revealed. Furthermore, a positive liner correlation between the UAE and CFT was found. Finally, the partial correlation coefficient adjusted for either the CFT or UAE indicated that both parameters directly correlated with the number of IVC injections administered during the 12 mo of follow-up. CONCLUSION: Generally, macular edema occurred in patients with severe NPDR, for whom the UAE is an independent risk predictor of DME. The baseline UAE and CFT predicted the treatment frequency of IVC injections administered in the first year for eyes with DME.

Optimal dietary copper requirements and relative bioavailability for weanling pigs fed either copper proteinate or tribasic copper chloride

Background: The objective of this study was to determine the effects of supplementing Cu on growth performance, Cu metabolism and Cu-related enzyme activities of weanling pigs fed diets with two different Cu sources, and to estimate optimal Cu requirements and relative bioavailability from these two sources for pigs.Methods: Weanling pigs were allocated to 14 treatments arranged factorially, including 6 added Cu levels(5, 10,20, 40, 80, 160 mg/kg), and 2 mineral sources(tribasic Cu chloride, TBCC and copper proteinate, Cu Pro), as well as one negative control(0 mg/kg added Cu level) and one maximum allowed level treatment(200 mg/kg TBCC) for the entire 38-d experiment. Growth performance, mineral status and enzyme activities were measured at the end of this study.Results: Increasing levels of Cu showed linear and quadratic responses(P < 0.01) for final BW, ADG and FCR regardless of the sources. Supplementation with TBCC(> 80 mg/kg) and Cu Pro(> 20 mg/kg) significantly decreased(P < 0.05) diarrhea incidence of weanling pigs. There were linear and quadratic increases(P < 0.01) in bile, hepatic,and intestinal Cu concentrations, fecal Cu contents, and plasma enzyme activities(alkaline phosphatase,ceruloplasmin, Cu, Zn-Superoxide dismutase(Cu/Zn SOD), and glutathione peroxidase), whereas plasma malondialdehyde decreased(P < 0.01) linearly and quadratically as dietary Cu level increased. Similarly, pigs fed Cu Pro absorbed and retained more Cu and excreted less Cu than those fed TBCC when supplemented 80 mg/kg and above. Optimal dietary Cu requirements for pigs from 28 to 66 d of age estimated based on fitted broken-line models(P < 0.05) of bile Cu, plasma Cu/Zn SOD and growth performance were 93–140 mg/kg from TBCC, and 63–98 mg/kg from Cu Pro accordingly. According to slope ratios from multiple linear regression, the bioavailability value of Cu Pro relative to TBCC(100%) was 156–263%(P < 0.01).Conclusion: The findings indicated that Cu recommendation from current NRC(5–6 mg/kg) was not sufficient to meet the high requirement of weanling pigs. Cu from Cu Pro was significantly more bioavailable to weanling pigs than TBCC in stimulating growth and enzyme activities, decreasing diarrhea frequency and fecal Cu contents to the environment.

EFFECTS OF REDUCED SALINITY ON OXYGEN CONSUMPTION AND AMMONIA-N EXCRETION OF CHLAMYS FARRERI

Effects of reduced salinity on the oxygen consumption rate (OCR) and the ammonia N excretion rate (AER) of scallop Chlamys farreri (3.2 5.9 cm in shell height, 0.147 1.635 g in soft tissue dry weight ) were studied in laboratory from March 21,1997 to April 16, 1997. Under the controlled conditions of reduced salinity from 31.5 to 15.0 and ambient temperature 17℃ and 23℃, the concentrations of dissolved oxygen and ammonia N were determined by the Winkle method and the hypobromite method, respectively. Results showed that with controlled reduced salinity, the mean values of the OCR were 2.17 mg/(g.h) at 17℃, and 2.86 mg/(g.h) at 23℃and that the mean values of the AER were 178.0 μg/(g.h) at 17℃ and 147.0 μg/(g.h) at 23℃. The OCR and the AER decreased with reducing salinity from 31.5 to 15.0 both at 17℃ and 23℃. The effects of reduced salinity on the OCR and the AER of scallop C. farreri could be represented by the allometric equation and the exponential equation, respectively.

Value of elastography point quantification in improving the diagnostic accuracy of early diabetic kidney disease

BACKGROUND Diabetic kidney disease(DKD)is a common complication of diabetes.The patient’s prognosis is poor once DKD progresses to advanced stage.Accurate diagnosis and timely treatment of early DKD are important for improving patient’s prognosis and reducing mortality.AIM To explore the value of elastography point quantification(ElastPQ)in improving the accuracy of early DKD diagnosis.METHODS A total of 69 patients with type 2 diabetes were recruited from Naval Military Medical University Affiliated Gongli Hospital.Patients were divided into early DKD group and medium DKD group according to pathological results and urinary albumin excretion rate(UAER).Another 40 patients with simple diabetes were included as the diabetes group.The baseline data,laboratory diagnostic indicators,and ultrasound indicators for each patient were recorded.The differences of the indicators in the three groups were compared.Multivariate logistic regression was used to analyze the influencing factors of the development from simple diabetes into early DKD and from early DKD into medium DKD.Receiver operating characteristic analyses of potential indicators in identifying early DKD and medium DKD,and early DKD and simple diabetes were established.RESULTS Multivariate logistic regression analysis showed that UAER(P<0.001),renocortical Young's Modulus(YM)(P<0.001),and renal parenchymal thickness(P=0.013)were the independent influencing factors of the development from early DKD into medium DKD.Diabetes duration(P=0.041),UAER(P=0.034),and renocortical YM(P=0.017)were the independent influencing factors of the development from simple diabetes into early DKD.Receiver operating characteristic analysis indicated that UAER,renocortical YM,and renal parenchymal thickness were accurate in identifying early DKD and medium DKD[all area under curve(AUC)>0.9].The accuracy of UAER(AUC=0.744),diabetes duration(AUC=0.757),and renocortical YM(AUC=0.782)for the diagnosis of early DKD and simple diabetes were limited.However,the combined diagnosis of UAER,diabetes duration,and renocortical YM was accurate in identifying early DKD and simple diabetes(AUC=0.906),which was significantly higher than any of the three indicators(all P<0.05).CONCLUSION ElastPQ is of great value in the diagnosis of early DKD.When combined with the diabetes duration and UAER,it is expected to diagnose accurately early DKD.

Combined fluorescence and electrochemical investigation on the binding interaction between organic acid and human serum albumin

Human serum albumin （HSA） is a plasma protein responsible for the binding and transport of fatty acids and a variety of exogenous chemicals such as drugs and environmental pollutants. Such binding plays a crucial role in determining the ADME （absorption, distribution, metabolism, and excretion） and bioavailability of the pollutants. The binding interaction between HSA and acetic acid （C2）, octanoic acid （C8） and dodecanoic acid （C12） has been investigated by the combination of site-specific fluorescent probe, tryptophan intrinsic fluorescence and tyrosine electrochemistry. For the study of the fatty acid interaction with the two drug-binding sites on HSA, two fluorescent probes, dansylamide and dansyl-L-proline were employed in the displacement measurements. Intrinsic fluorescence of tryptophan in HSA was monitored upon addition of the fatty acids into HSA. Electrocatalyzed response of the tyrosine residues in HSA by a redox mediator was used to investigate the binding interaction. Qualitatively, observations from these three approaches were very similar. HSA did not show any change in the fluorescence and electrochemical experiments after mixing with C2, suggesting there is no significant interaction with the short-chain fatty acid. For C8, the measured signal dropped in a single-exponential mode, indicating an independent and non-cooperative binding. The calculated association constant and binding ratio were 3.1 × 10^6 L/mol and 1 with drug binding Site Ⅰ, 1.1 × 107 L/mol and 1 with Site Ⅱ, and 7.0× 0^4 L/mol and 4 with the tryptophan site, respectively. The measurements with C12 displayed multiple phases of fluorescence change, suggesting cooperativity and allosteric effect of the C12 binding. These results correlate well with those obtained by the established methods, and validate the new approach as a viable tool to study the interactions of environmental pollutants with biological molecules.

Effect of Eurycoma longifolia stem extract on uric acid excretion in hyperuricemia mice

OBJECTIVE Eurycoma longifolia is a tropical medicinal plant belonging to Simaroubaceae distributed in South East Asia.The aim of this study is to explore the effect and mechanism of E.longifolia stem 70%ethanol extract(EL)and its active com⁃poundson uric acid excretion.METHODS Potassium oxonate(PO)induced hyperuricemia rats and adenine-PO induced hyperuricemia mouse model were used to evaluate the effects of EL.Ultra Performance Liquid Chromatography was used to determine the levels of plasma or serum uric acid and creatinine.Hematoxylin-eosin staining was applied to observe kidney pathological changes,Western blot⁃ting was applied to detect protein expression levels of uric acid transporters.Effects of constituents on urate uptake were tested in hU⁃RAT1-expressing HEK293T cells.RESULTS EL significantly reduced serum and plasma uric acid levels at dosages of 100,200 and 400 mg·kg^-1 in hyperuricemia rats and mice,and increased the clearance rate of uric acid and creatinine,improved therenal pathological injury.The protein expression levels of urate reabsorption transporter 1(URAT1)and glucose transporter 9 were down-regulated while sodium-dependent phosphate transporter 1 and ATP-binding cassette transporter G2 were up-regulated in the kidney after EL treat⁃ment.The diterpenes(50μmol·L^-1)isolated from EL showed inhibitory effects on urate uptake in hURAT1-expressing HEK293T cells,and the effect of eurycomanol was further confirmed in vivo.CONCLUSION EL significantly reduced blood uric acid levels and prevented pathological changes of kidney in PO induced hyperuricemia animal model,improved renal urate transports.We partly clarified the mechanism was related to suppressing effect of URAT1 by diterpene in EL.This study is the first to demonstrate that EL plays a role in hyperuricemia by promoting renal uric acid excretion.

Gluten immunogenic peptide excretion detects dietary transgressions in treated celiac disease patients

BACKGROUND Life-long removal of gluten from the diet is currently the only way to manage celiac disease(CeD). Until now, no objective test has proven useful to objectively detect ingested gluten in clinical practice. Recently, tests that determine consumption of gluten by assessing excretion of gluten immunogenic peptides(GIP) in stool and urine have been developed. Their utility, in comparison with conventional dietary and analytical follow-up strategies, has not been fully established.AIM To assess the performance of enzyme-linked immunosorbent assay(ELISA) and point-of-care tests(PoCTs) for GIP excretion in CeD patients on gluten-free diet(GFD).METHODS We conducted an observational, prospective, cross-sectional study in patients following a GFD for at least two years. Using the Gastrointestinal Symptom Rating Scale questionnaire, patients were classified at enrollment as asymptomatic or symptomatic. Gluten consumption was assessed twice by 3-d dietary recall and GIP excretion(by ELISA in stool and PoCTs(commercial kits for stool and urine) in two consecutive samples. These samples and dietary reports were obtained 10 day apart one from the other. Patients were encouraged to follow their usual GFD during the study period.RESULTS Forty-four patients were enrolled, of which 19(43.2%) were symptomatic despite being on a GFD. Overall, 83 sets of stool and/or urine samples were collected.Eleven out of 44 patients(25.0%) had at least one positive GIP test. The occurrence of at least one positive test was 32% in asymptomatic patients compared with 15.8% in symptomatic patients. GIP was concordant with dietary reports in 65.9% of cases(Cohen′s kappa: 0.317). PoCT detected dietary indiscretions. Both ELISA and PoCT in stool were concordant(concomitantly positive or negative) in 67 out of 74(90.5%) samples. Excretion of GIP was detected in 7(8.4%) stool and/or urine samples from patients considered to be strictly compliant with the GFD by dietary reports.CONCLUSION GIP detects dietary transgressions in patients on long-term GFD, irrespective of the presence of symptoms. PoCT for GIP detection constitutes a simple homebased method for self-assessment of dietary indiscretions.

A Participant-assigned Interventional Research of Precesarean Internal Iliac Artery Balloon Catheterization for Managing Intraoperative Hemorrhage of Placenta Previa and Placenta Accreta Spectrum Disorders After Cesarean Section

Placenta accreta spectrum disorder (PASD) and placenta previa (PP) are two of the mosthideous obstetric complications which are usually associated with a history of cesarean section(CS). Moreover, women with PASD, PP and/or a cesarean scarred uterus are more likely to haveadverse pregnancy outcomes, including blood transfusion, hysterectomy, pelvic organs damage,postpartum hemorhage, disseminated intravascular coagulation, multi-organ dysfunction syndromeand even maternal or fetal death. This study aimed to investigate the efficacy of precesareaninternal iliac artery balloon catheterization (BC) for managing severe hemorhage caused by PASDand PP with a history of CS. This participant-assigned interventional study was conducted inTongji Hospital. We recruited 128 women with suspected PASD, PP and a history of CS. Womenin the BC group accepted precesarean BC of bilateral internal iliac arteries before the scheduledcesarean delivery. Women in the control group underwent a conventional cesarean delivery.Intraoperative hemorrhage, transfusion volume, radiation dose, exposure time, complicationsand neonatal outcomes were discussed. There were significant differences in calculated bloodloss (CBL) between BC group and control group (1015.0±144.9 vs. 1467.0±171.0 mL, P=0.04).Precesarean BC could reduce intraoperative red blood cell (RBC) transfusion as compared withcontrol group (799.5±136.1 vs. 1286.0±161.6 mL, P=0.02) and lessen the rate of using bloodproducts (57.1% vs. 76.4%, P=0.02). The incidence of hysterectomy was also lower in BC groupthan in control group. Postpartum outcomes showed no significant differences between the twogroups, except that postoperation hospitalization was longer in BC group than in control group(6.7±0.4 vs. 5.8±0.2 days, P=0.03). Precesarean BC of internal iliac artery is an effective methodfor managing severe hemorrhage caused by PASD and PP with a cesarean scarred uterus, as it couldreduce intraoperative blood loss, lessen intraoperative RBC transfusions and potentially decreasehysterectomies.