Epsin1-mediated exosomal sorting of Dll4 modulates the tubular-macrophage crosstalk in diabetic nephropathy.

Tubular epithelial cells (TECs) play critical roles in the development of diabetic nephropathy (DN), and can activate macrophages through the secretion of exosomes. However, the mechanism(s) of TEC-exosomes in macrophage activation under DN remains unknown. By mass spectrometry, 1,644 differentially expressed proteins, especially Dll4, were detected in the urine exosomes of DN patients compared with controls, which was confirmed by western blot assay. Elevated Epsin1 and Dll4/N1ICD expression was observed in kidney tissues in both DN patients and db/db mice and was positively associated with tubulointerstitial damage. Exosomes from high glucose (HG)-treated tubular cells (HK-2) with Epsin1 knockdown (KD) ameliorated macrophage activation, TNF-α, and IL-6 expression, and tubulointerstitial damage in C57BL/6 mice in vivo. In an in vitro study, enriched Dll4 was confirmed in HK-2 cells stimulated with HG, which was captured by THP-1 cells and promoted M1 macrophage activation. In addition, Epsin1 modulated the content of Dll4 in TEC-exosomes stimulated with HG. TEC-exosomes with Epsin1-KD significantly inhibited N1ICD activation and iNOS expression in THP-1 cells compared with incubation with HG alone. These findings suggested that Epsin1 could modulate tubular-macrophage crosstalk in DN by mediating exosomal sorting of Dll4 and Notch1 activation.

Selection of suitable reference genes for qPCR normalization in different developmental stages of Oenanthe javanica.

Gene expression analysis is widely used to unravel molecular regulatory mechanisms and identify key genes in plants. Appropriate reference gene is an important prerequisite to ensure the accuracy and reliability of qPCR analysis results. Water dropwort is a plant of the Oenanthe genus in the Apiaceae family, which has high economic benefits. However, the underlying molecular regulatory mechanisms in the growth and development of water dropwort have not been fully understood and the appropriate reference genes in different developmental stages of water dropwort not yet reported. In this study, 10 candidate reference genes (ACTIN, PP2A, SAND, EF-1α, GAPDH, UBQ, MIP, TBP, RPS-18, eIF-4α) were identified and cloned from Oenanthe javanica. The qPCR primers of candidate reference genes were designed and verified. Four statistical algorithms, geNorm, NormFinder, BestKeeper and RefFinder were used to evaluate the expression stability of 10 candidate reference genes in different developmental stages of water dropwort. The results showed that TBP and UBQ were the most stable genes in different developmental stages of water dropwort, while GAPDH was the most unstable gene. The normalization of EXP1 genes at different developmental stages further confirmed the reliability of internal reference genes. The results of this study provide a theoretical basis for selecting appropriate internal reference genes in different developmental stages of water dropwort. This study also provides technical support and reliable basis for the expression analysis of key genes in different developmental stages of water dropwort.

Genetic spectrum of CAKUT and risk factors for kidney failure: a pediatric multicenter cohort study.

BACKGROUND: Congenital anomalies of the kidney and urinary tracts (CAKUT) are the leading cause of kidney failure in children with phenotypic and genotypic heterogeneity. Our objective was to describe the genetic spectrum and identify the risk factors for kidney failure in children with CAKUT. METHODS: Clinical and genetic data were derived from a multicenter network (Chinese Children Genetic Kidney Disease Database, CCGKDD) and the Chigene database. A total of 925 children with CAKUT who underwent genetic testing from 2014 to 2020 across China were studied. Data for a total of 584 children wereobtained from the CCGKDD, including longitudinal data regarding kidney function. The risk factors for kidney failure were determined by the Kaplan-Meier method and Cox proportional hazards models. RESULTS: A genetic diagnosis was established in 96 out of 925 (10.3%) children, including 72 (8%) with monogenic variants, 20 (2%) with copy number variants (CNVs), and 4 (0.4%)with major chromosomal anomalies. Patients with skeletal abnormalities were more likely to have large CNVs or abnormal karyotypes than monogenic variants. Eighty-two patients from the CCGKDD progressed to kidney failure at a median age of 13.0 (95% confidence interval, 12.4-13.6) years, and twenty-four were genetically diagnosed with variants of PAX2, TNXB, EYA1, HNF1B and GATA3 or the 48, XXYY karyotype. The multivariate analysis indicated that solitary kidney, posterior urethral valves, bilateral hypodysplasia, the presence of certain variants and premature birth were independent prognostic factors. CONCLUSIONS: The genetic spectrum of CAKUT varies among different subphenotypes. The identified factors indicate areas that require special attention.

Epac activation ameliorates tubulointerstitial inflammation in diabetic nephropathy.

Tubulointerstitial inflammation plays an important role in the progression of diabetic nephropathy (DN), and tubular epithelial cells (TECs) are crucial promoters of the inflammatory cascade. Exchange protein activated by cAMP (Epac) has been shown to suppress the angiotensin II (Ang-II)-induced release of inflammatory cytokines in tubular cells. However, the role of Epac in TEC-mediated tubulointerstitial inflammation in DN remains unknown. We found that administering the Epac agonist 8-pCPT-2'-O-Me-cAMP (8-O-cAMP) to db/db mice inhibited tubulointerstitial inflammation characterized by macrophage infiltration and increased inflammatory cytokine release and consequently alleviated tubulointerstitial fibrosis in the kidney. Furthermore, 8-O-cAMP administration restored CCAAT/enhancer binding protein ß (C/EBP-ß) expression and further upregulated the expression of Suppressor of cytokine signaling 3 (SOCS3), while inhibiting p-STAT3, MCP-1, IL-6, and TNF-α expression in the kidney cortex in db/db mice. And in vitro study showed that macrophage migration and MCP-1 expression induced by high glucose (HG, 30 mM) were notably reduced by 8-O-cAMP in human renal proximal tubule epithelial (HK-2) cells. In addition, 8-O-cAMP treatment restored C/EBP-ß expression in HK-2 cells and promoted C/EBP-ß translocation to the nucleus, where it transcriptionally upregulated SOCS3 expression, subsequently inhibiting STAT3 phosphorylation. Under HG conditions, siRNA-mediated knockdown of C/EBP-ß or SOCS3 in HK-2 cells partially blocked the inhibitory effect of Epac activation on the release of MCP-1. In contrast, SOCS3 overexpression inhibited HG-induced activation of STAT3 and MCP-1 expression in HK-2 cells. These findings indicate that Epac activation via 8-O-cAMP ameliorates tubulointerstitial inflammation in DN through the C/EBP-ß/SOCS3/STAT3 pathway.

Responsible genes in children with primary vesicoureteral reflux: findings from the Chinese Children Genetic Kidney Disease Database.

BACKGROUND: Primary vesicoureteral reflux (VUR) is a common congenital anomaly of the kidney and urinary tract (CAKUT) in childhood. The present study identified the possible genetic contributions to primary VUR in children. METHODS: Patients with primary VUR were enrolled and analysed based on a national multi-center registration network (Chinese Children Genetic Kidney Disease Database, CCGKDD) that covered 23 different provinces/regions in China from 2014 to 2019. Genetic causes were sought using whole-exome sequencing (WES) or targeted-exome sequencing. RESULTS: A total of 379 unrelated patients (male: female 219:160) with primary VUR were recruited. Sixty-four (16.9%) children had extrarenal manifestations, and 165 (43.5%) patients showed the coexistence of other CAKUT phenotypes. Eighty-eight patient (23.2%) exhibited impaired renal function at their last visit, and 18 of them (20.5%) developed ESRD at the median age of 7.0 (IQR 0.9-11.4) years. A monogenic cause was identified in 28 patients (7.39%). These genes included PAX2 (n = 4), TNXB (n = 3), GATA3 (n = 3), SLIT2 (n = 3), ROBO2 (n = 2), TBX18 (n = 2), and the other 11 genes (one gene for each patient). There was a significant difference in the rate of gene mutations between patients with or without extrarenal complications (14.1% vs. 6%, P = 0.035). The frequency of genetic abnormality was not statistically significant based on the coexistence of another CAKUT (9.6% vs. 5.6%, P = 0.139, Chi-square test) and the grade of reflux (9.4% vs. 6.7%, P = 0.429). Kaplan-Meier survival curve showed that the presence of genetic mutations did affect renal survival (Log-rank test, P = 0.01). PAX2 mutation carriers (HR 5.1, 95% CI 1.3-20.0; P = 0.02) and TNXB mutation carriers (HR 20.3, 95% CI 2.4-168.7; P = 0.01) were associated with increased risk of progression to ESRD. CONCLUSIONS: PAX2, TNXB, GATA3 and SLIT2 were the main underlying monogenic causes and accounted for up to 46.4% of monogenic VUR. Extrarenal complications and renal function were significantly related to the findings of genetic factors in children with primary VUR. Like other types of CAKUT, several genes may be responsible for isolated VUR.

A review on the efficacy and mechanism of action of Shenkang injection against chronic kidney disease.

Chronic kidney disease (CKD) is one of the most common conditions which significantly increases the risk for serious health outcomes. Epidemiological investigations have shown that CKD has become a serious global health problem. At present, there are no treatments for CKD, thus the need for an effective and safe treatment for this condition. Shenkang Injection (SKI), which is an herbal medication in Chinese Medicine, has been used in the management and treatment of CKD and has achieved favorable therapeutic effects. The purpose of this paper is to review the clinical efficacy, mechanism of action, and safety profile of SKI when used in CKD, and to provide comprehensive potential evidence for its clinical application.

Functional analysis of UGT201D3 associated with abamectin resistance in Tetranychus cinnabarinus (Boisduval).

Uridine diphosphate (UDP)-glycosyltransferases (UGTs) are widely distributed within living organisms and share roles in biotransformation of various lipophilic endo- and xenobiotics with activated UDP sugars. In this study, it was found that the activity of UGTs in abamectin-resistant (AbR) strain was significantly higher (2.35-fold) than that in susceptible strain (SS) of Tetranychus cinnabarinus. Further analysis showed that 5-nitrouracil, the inhibitor of UGTs, could enhance the lethal effect of abamectin on mites. From the previous microarray results, we found an UGT gene (UGT201D3) overexpressed in AbR strain. Quantitative PCR analysis showed that UGT201D3 was highly expressed and more inducible with abamectin exposure in the AbR strain. After silencing the transcription of UGT201D3, the activity of UGTs was decreased and the susceptibility to abamectin was increased in AbR strain whereas it was not in SS. Furthermore, UGT201D3 gene was then successfully expressed in Escherichia coli. The recombinant UGT201D3 exhibited α-naphthol activity (2.81 ± 0.43 nmol/mg protein/min), and the enzyme activity could be inhibited by abamectin (inhibitory concentration at 50%: 57.50 ± 3.54 µmol/L). High-performance liquid chromatography analysis demonstrated that the recombinant UGT201D3 could effectively deplete abamectin (15.77% ± 3.72%) incubating with 150 µg protein for 6 h. These results provided direct evidence that UGT201D3 was involved in abamectin resistance in T. cinnabarinus.

Tetra-kis(µ(2)-2-phen-oxy-propionato)-κO,O':O';κO:O,O';κO:O'-bis-[(1,10-phenanthroline-κN,N')(2-phen-oxy-propionato-κO,O')cerium(III)].

In the centrosymmetric binuclear title complex, [Ce(2)(C(9)H(9)O(3))(6)(C(12)H(8)N(2))(2)], the two Ce(III) ions are linked by four 2-phen-oxy-propionate groups in bi- and tridentate bridging modes. Each Ce(III) ion is nine-coordinated by one 1,10-phenanthroline mol-ecule, two O atoms from a chelating carboxyl-ate, two O atoms derived from a µ(3)-carboxylate and two O atoms derived from two µ(2)-carboxylate ligands in a distorted CeN(2)O(7) monocapped square-anti-prismatic geometry.

Tetra-kis(µ(2)-2-phen-oxy-propionato)-κO,O':O';κO:O,O';κO:O'-bis-[(1,10-phenanthroline-κN,N')(2-phen-oxy-propionato-κO,O')gadolinium(III)].

In the centrosymmetric binuclear title complex, [Gd(2)(C(9)H(9)O(3))(6)(C(12)H(8)N(2))(2)], the two Gd(III) ions are linked by four 2-phen-oxy-propionate (L) groups in bi- and tridentate bridging modes. Each Gd(III) ion is nine-coordinated by one 1,10-phenanthroline mol-ecule, one bidentate chelating carboxyl-ate group and four bridging carboxyl-ate groups in a distorted GdN(2)O(7) monocapped square-anti-prismatic geometry.

Bis(µ-2-phen-oxy-propionato-κO:O')bis-[(1,10-phenanthroline-κN,N')bis-(2-phen-oxy-propionato-κO,O')samarium(III)].

The dimeric title compound, [Sm(2)(C(9)H(9)O(3))(6)(C(12)H(8)N(2))(2)], is centrosymmetric and is composed of six 2-phen-oxy-propionate anions and two 1,10-phenanthroline ligands. The Sm(III) atom is coordinated by two O atoms from two bridging anions, four O atoms from two chelating anions and the N atoms of the N-heterocycle in a distorted dodeca-hedral geometry.

Tetra-kis(µ(2)-2-phen-oxy-propionato)-κO,O':O';κO:O,O';κO:O'-bis-[(1,10-phenanthroline-κN,N')(2-phen-oxy-propionato-κO,O')lanthanum(III)].

In the centrosymmetric binuclear title complex, [La(2)(C(9)H(9)O(3))(6)(C(12)H(8)N(2))(2)], the two La(III) ions are linked by four 2-phen-oxy-propionate (L) groups in bi- and tridentate bridging modes. Each La(III) ion is nine-coordinated by one 1,10-phenanthroline mol-ecule, one bidentate chelating carboxyl-ate group and four bridging carboxyl-ate groups in a distorted LaN(2)O(7) monocapped square-anti-prismatic geometry.

Bis(µ(2)-2-phen-oxy-propionato-κO:O')bis-[(1,10-phenanthroline-κN,N')bis-(2-phen-oxy-propionato-κO,O')ytterbium(III)].

In the centrosymmetric binuclear title complex, [Yb(2)(C(9)H(9)O(3))(6)(C(12)H(8)N(2))(2)], the two Yb(III) ions are linked by two 2-phen-oxy-propionate (L) groups in a bidentate bridging mode. Each Yb(III) ion is eight-coordinated by two O atoms from two bridging L ligands, four O atoms from two chelating L groups and two N atoms from one chelating phen mol-ecule in a distorted YbN(2)O(6) dodeca-hedral geometry.

Tetra-kis(µ-2-phen-oxy-propionato)-κO,O':O';κO:O,O';κO:O'-bis-[(1,10-phenanthroline-κN,N')(2-phen-oxy-propionato-κO,O')holmium(III)].

The title compound, [Ho(2)(C(9)H(9)O(3))(6)(C(12)H(8)N(2))(2)], lies about a centre of symmetry and is comprised of six 2-phen-oxy-propionate (POPA) anions and two 1,10-phenanthroline (phen) ligands. The two Ho(III) ions are linked by four POPA groups utilizing both bi- and tridentate bridging modes to form an inversion-symmetric dimer. Each Ho(III) ion is nine-coordinate, with a chelating 1,10-phenanthroline mol-ecule, one bidentate chelating carboxyl-ate group, two bidentate bridging carboxyl-ate groups and two tridentate bridging carboxyl-ate groups in a distorted mono-capped square anti-prism geometry. There are weak π-π aromatic inter-actions between the phen groups and aromatic rings of the POPA ligands [centroid-centroid distance = 3.829 (1) Å].

Tetra-kis(µ-2-phen-oxy-propionato)-κO,O':O';κO:O,O';κO:O'-bis-[(1,10-phenanthroline-κN,N')(2-phen-oxy-propionato-κO,O')terbium(III)].

In the centrosymmetric binuclear title complex, [Tb(2)(C(9)H(9)O(3))(6)(C(12)H(8)N(2))(2)], the two Tb(III) ions are linked by four 2-phen-oxy-propionate (L) groups through their bi- and tridentate bridging modes. Each Tb(III) ion is nine-coordinated by one 1,10-phenanthroline mol-ecule, one bidentate carboxyl-ate group and four bridging carboxyl-ate groups in a distorted TbN(2)O(7) monocapped square-anti-prismatic geometry.

Tetra-kis(µ-2-phen-oxy-propionato)-κO,O':O';κO:O,O',κO:O'-bis-[(1,10-phenanthroline-κN,N')(2-phen-oxy-propionato-κO,O')dysprosium(III)].

In the centrosymmetric binuclear title complex, [Dy(2)(C(9)H(9)O(3))(6)(C(12)H(8)N(2))(2)], the two Dy(III) ions are linked by four 2-phen-oxy-propionate (L) groups through their bi- and tridentate bridging modes. Each Dy(III) ion is nine-coordinated by one 1,10-phenanthroline mol-ecule, one bidentate carboxyl-ate group and four bridging carboxyl-ate groups in a distorted DyN(2)O(7) monocapped square-anti-prismatic geometry. The title compound is isotypic with its terbium-containing analogue.

Tetra-kis(µ-2-phen-oxy-propionato)-κO,O':O';κO:O,O',κO:O'-bis-[(1,10-phenanthroline-κN,N')(2-phen-oxy-propionato-κO,O')praseodymium(III)].

In the centrosymmetric binuclear title complex, [Pr(2)(C(9)H(9)O(3))(6)(C(12)H(8)N(2))(2)], the two Pr(III) ions are linked by four 2-phen-oxy-propionate (L) groups through their bi- and tridentate bridging modes. Each Pr(III) ion is nine-coordinated by one 1,10-phenanthroline mol-ecule, one bidentate carboxyl-ate group and four bridging carboxyl-ate groups in a distorted PrN(2)O(7) monocapped square-anti-prismatic geometry. The title compound is isotypic with its terbium- and dysprosium-containing analogues.

[µ-2-(4-Hy-droxy-phen-yl)acetato]-κO:O,O';κO,O':O'-bis-{aqua-(4,4'-bi-pyridine-κN)bis-[2-(4-hy-droxy-phen-yl)acetato-κO,O']holmium(III)} mono-hydrate.

In the title dinuclear complex, [Ho(2)(C(8)H(7)O(3))(6)(C(10)H(8)N(2))(2)(H(2)O)(2)]·H(2)O, each of the two independent Ho(III) ions is coordinated by eight O atoms from four 4-hy-droxy-phenyl-acetate (HPAA) ligands and a water mol-ecule, and one N atom from a 4,4'-bipyridine (bipy) ligand in a distorted tricapped trigonal-prismatic geometry. The HPAA ligands are coordinated in bis-chelate, bridging and bridging tridentate modes. In the crystal, O-H⋯O and O-H⋯N hydrogen bonds link the mol-ecules into a three-dimensional network.

Poly[(µ(2)-4,4'-bipyridine-κN:N')bis-(µ(2)-2-phen-oxy-propionato-κO:O')cobalt(II)].

In the polymeric title compound, [Co(C(9)H(9)O(3))(2)(C(10)H(8)N(2))](n), the Co(II) ion is located on a twofold rotation axis and is six-coordinated by two N atoms from two 4,4'-bipyridine (4,4'-bipy) ligands in axial positions and by four O atoms from four 2-phen-oxy-propionate (POPA) anions in equatorial positions, defining a slightly distorted octa-hedral geometry. The carboxyl-ate group of the POPA anion displays a bis-monodentate mode, linking pairs of Co(II) ions into a chain parallel to [001]. Adjacent chains are connected in a perpendicular manner through 4,4'-bipy ligands into layers parallel to (100). The 4,4'-bipy ligand is likewise located on a twofold rotation axis, with a dihedral angle between the two pyridine rings of 57.05 (7)°. C-H⋯O hydrogen-bonding inter-actions are present within the layers. π-π stacking inter-actions between the POPA benzene rings of neighbouring layers [centroid-to-centroid distance = 3.976 (3) Šand plane-to-plane distance = 3.618 (3) Å] stabilize the packing of the structure.

Tetra-kis(µ-3,4-dimeth-oxy-phenyl-acetato)-bis-[(3,4-dimeth-oxy-phenyl-acetato)(1,10-phenanthroline)holmium(III)].

In the centrosymmetric title compound, [Ho(2)(C(10)H(11)O(4))(6)(C(12)H(8)N(2))(2)], the Ho(III) atom is nine-coordinated by seven O atoms from the 3,4-dimeth-oxy-phenyl-acetate (L) anions and two N atoms from a 1,10-phenanthroline (phen) mol-ecule. The L ligands are coordinated to the Ho(III) ions in three modes: chelating, bridging and bridging-tridentate. Intra-molecular C-H⋯O inter-actions occur. The crystal packing is stabilized by inter-molecular C-H⋯O inter-actions and weak aromatic π-π inter-actions between phen mol-ecules and the aromatic rings of the L ligands [centroid-centroid distance = 3.821 (2) Å].

(Methanol-κO)bis-{2-meth-oxy-6-[(4-methyl-phen-yl)iminiometh-yl]phenolato-κO,O'}tris-(nitrato-κO,O')lanthanum(III).

The asymmetric unit of title compound, [La(NO(3))(3)(C(15)H(15)NO(2))(2)(CH(3)OH)], consists of two Schiff base 2-meth-oxy-6-[(4-methyl-phen-yl)iminiometh-yl]phenolato (HL) ligands, three independent nitrate anions and one methanol mol-ecule coordinated to La(III). The coordination environment of the La(III) ion is formed by eleven O atoms. Three bidentate nitrate anions coordinate to the La(III) ion, while two HL ligands chelate the metal center with O atoms from the phenolate and meth-oxy groups. The HL ligands are zwitterionic, with protonated imine N atoms. The coordination sphere is completed by one methanol mol-ecule. The protonated imine N atoms are involved in intra-molecular N-H⋯O hydrogen bonds with the phen-oxy groups and nitrate ligands. One O atom of one nitrate group is disordered over two sites of equal occupancy.