Characteristics of innate and adaptive immune disorder in IgA nephropathy based on integrated bioinformatics.

BACKGROUND: Our study aims to investigate the immunological pathogenesis underlying immunoglobulin A nephropathy (IgAN) and explore potential biomarkers for IgAN diagnosis. MATERIALS AND METHODS: Differentially expressed genes (DEGs) of formalin-fixed and paraffin-embedded (FFPE) samples were screened between IgAN patients and healthy people based on GSE115857. Gene oncology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Set Enrichment Analysis (GSEA) enrichment was performed to identify related biological processes and pathways. CIBERSORT was utilized to seek the relationship of immune cell infiltration with IgAN. Finally, the expression of paraoxonase 2 (PON2) related to innate immune response was verified in FFPE samples of minimal change disease and IgAN patients by immunohistochemistry and PAS staining. RESULTS: 25 down-regulated genes and 12 up-regulated genes were identified in IgAN patients, which mainly responded to endothelial cell proliferation, inflammatory response, and angiogenesis. Toll-like receptor signaling pathway and Epstein-Barr virus (EBV) infection might be involved in IgAN pathogenesis. In addition, the infiltration of macrophages M0, naïve B cells, and follicular helper T (Tfh) cells was positively correlated in IgAN patients. Macrophages M1 and M2 infiltration were up-regulated in IgAN patients, which indicated that innate immune response was closely associated with IgAN. Besides, the results of immunohistochemistry showed that PON2 was obviously positively expressed in acute and chronic lesions of IgAN patients. CONCLUSION: In addition to abnormalities in the adaptive immune response, macrophages M1/M2 and innate immune disorder may participate in IgAN pathogenesis. PON2 may become the feasible targets for further investigation of IgAN.

Employing Pd nanoparticles decorated on halloysite nanotube/carbon composite for electrochemical aptasensing of HER2 in breast cancer patients.

An effective biosensing platform is described based on halloysite nanotube/carbon composite decorated with Pd nanoparticles (HNT/C@Pd NPs). A novel electrochemical aptasensor was designed using the proposed nano-platform to determine human epidermal growth factor receptor 2 (HER2), a breast cancer biomarker. Inherently, aptasensing interfaces provide high sensitivity and selectivity for tumor markers owing to the high specific surface area of HNT/C and good conductivity stems from deposition of Pd NPs into HNT/C composite. With a correlation coefficient of 0.996, the electrochemical aptasensor demonstrated a wide linear range from 0.03 ng/mL to 9 ng/mL. The limit of detection (LOD) of the established assay was 8 pg/mL based on S/N = 3 method. Further, the designed biosensor demonstrated acceptable selectivity, good reproducibility, and high stability. The applicability of the impedimetric sensor in human serum samples was also examined and compared to enzyme-linked immunosorbent assay (ELISA) assay (p-value >0.05). Based on the results, it was found that the proposed methodology can be used in quantification of breast cancer markers for early diagnosis and treatment.

Annexin A2: The diversity of pathological effects in tumorigenesis and immune response.

Annexin A2 (ANXA2) is widely used as a marker in a variety of tumors. By regulating multiple signal pathways, ANXA2 promotes the epithelial-mesenchymal transition, which can cause tumorigenesis and accelerate thymus degeneration. The elevated ANXA2 heterotetramer facilitates the production of plasmin, which participates in pathophysiologic processes such as tumor cell invasion and metastasis, bleeding diseases, angiogenesis, inducing the expression of inflammatory factors. In addition, the ANXA2 on the cell membrane mediates immune response via its interaction with surface proteins of pathogens, C1q, toll-like receptor 2, anti-dsDNA antibodies and immunoglobulins. Nuclear ANXA2 plays a role as part of a primer recognition protein complex that enhances DNA synthesis and cells proliferation by acting on the G1-S phase of the cell. ANXA2 reduction leads to the inhibition of invasion and metastasis in multiple tumor cells, bleeding complications in acute promyelocytic leukemia, retinal angiogenesis, autoimmunity response and tumor drug resistance. In this review, we provide an update on the pathological effects of ANXA2 in both tumorigenesis and the immune response. We highlight ANXA2 as a critical protein in numerous malignancies and the immune host response.

Correlation of urine protein/creatinine ratios to 24-h urinary protein for quantitating proteinuria in children.

BACKGROUND: The purpose of this study was to evaluate the correlation and consistency between urine protein/creatinine ratio (UPCR) and 24-h urine protein (24HUPr) in children, and to determine cutoff values of UPCR relative to 24HUPr at 100 mg/m2/d (≥ 100 mg/m2/d as pathological proteinuria) and 1000 mg/m2/d (≥ 1000 mg/m2/d as nephrotic-range proteinuria). METHODS: Three hundred sixty-six children were enrolled, including 81 controls, 109 with Henoch-Schönlein purpura nephritis, 167 with nephrotic syndrome, 5 with IgA nephropathy, and 4 with lupus nephritis. Patients were divided into three groups: normal group; non-nephrotic-range proteinuria group; nephrotic-range proteinuria group. The cutoff values of UPCR in predicting the different levels of proteinuria were determined using ROC curve analysis. RESULTS: UPCR was positively correlated with 24HUPr (r = 0.915, p < 0.01). Bland-Altman diagrams showed that UPCR and 24HUPr had good consistency, and > 95% spots of UPCR and 24HUPr were within 95% confidence intervals. Relative to 24HUPr at 100 mg/m2/d, the cutoff value of UPCR (0.18 g/g Cr) had the highest sensitivity (94%) and specificity (98.8%) which is close to 0.2 g/g Cr as proposed by the American College of Rheumatology. Relative to 24HUPr at 1000 mg/m2/d, the cutoff value of UPCR (2.09 g/g Cr) had the highest sensitivity (92.1%) and specificity (92.1%) which is close to the 2.0 g/g Cr proposed in KDIGO guidelines. CONCLUSIONS: UPCR showed strong correlation and consistency with 24HUPr for evaluating levels of proteinuria in children. UPCR < 0.2 g/g Cr can be considered a criterion for normal-range proteinuria. Instead of 24HUPr ≥ 1000 mg/m2/d, UPCR ≥ 2.0 g/g Cr can be considered a criterion for nephrotic-range proteinuria or nephrotic syndrome in children.

Diverse nodule bacteria were associated with Astragalus species in arid region of northwestern China.

The legume species of Astragalus as traditional Chinese medicine source and environmental protection plants showed an extensive distribution in the arid region of northwestern China. However, few rhizobia associating with Astragalus have been investigated in this region so far. In this study, 78 endophytic bacteria were isolated from root nodules of 12 Astragalus species and characterized by the PCR-RFLP of 16S rRNA gene and symbiotic genes together with the phylogenetic analysis. Results showed that the majority (53%) of isolates are non-nodulating Agrobacterium sp. and the rest are Mesorhizobium genomic species (41%), Ensifer spp. and Rhizobium gallicum (6%), respectively. Mesorhizobium genomic species are broadly distributed in the Astragalus symbioses and most of them share similar symbiotic genes. It seems that horizontal gene transfer occurred frequently among different genomic species independent of their original hosts and sites. Astragalus adsurgens is nodulated by a widely range of rhizobial species in the nodulation test, revealing that it could play an important role in diversification of Astragalus symbionts and that might be a reason for its wide adaptation to diverse environments.

Urinary matrix metalloproteinase-7 is a sensitive biomarker to evaluate renal tubular injury in patients with minimal change disease and focal segmental glomerulosclerosis.

Objective: To explore the advantages of urinary matrix metalloproteinase-7 (MMP-7) in evaluating renal tubular injury in minimal change disease (MCD) and focal segmental glomerulosclerosis (FSGS) patients compared with urinary cystatin C (CysC) and retinol-binding protein (RBP). Methods: Serum and urine samples were collected from 20 healthy volunteers, and 40 MCD and 20 FSGS patients. Serum and urinary MMP-7 levels were measured by enzyme-linked immunosorbent assay. Urinary total protein, CysC and RBP levels were measured by automatic specific protein analyzer and compared with urinary creatinine level for calibration. The renal tissue serial sections were stained by MMP-7 immunohistochemistry and periodic acid-Schiff. Results: Under light microscopy, MMP-7 granular weak positive expression was showed sporadically in the cytoplasm of a few renal tubular epithelial cells without obvious morphological changes in MCD patients, and MMP-7-positive expression was observed in the cytoplasm of some renal tubular epithelial cells in FSGS patients. There was no significant difference in serum MMP-7 level among the three groups. Compared with the control group, the urinary MMP-7 level in MCD patients was higher, but urinary CysC and RBP levels were not increased significantly. Compared with the control group and MCD patients, urinary MMP-7, CysC and RBP levels in FSGS patients were upregulated significantly. Conclusions: Urinary MMP-7 could not only evaluate the mild renal tubular epithelial cells injury in MCD patients with massive proteinuria, but also evaluate the continuous renal tubular epithelial cells injury in FSGS patients.

To investigate the correlation between normal fetal biventricular myocardial function and gestational age using velocity vector imaging.

Objective: The aim of this study was to evaluate the left and right ventricular segmental and global myocardial function of normal fetuses using velocity vector imaging and explore the correlation between global myocardial function parameters and gestational age. Methods: A total of 127 normal fetuses were selected and divided into five groups according to gestational age for the measurement of their left and right ventricular segmental and global velocity, strain, and strain rate. This study also explored the change trend in the global myocardial function parameters at different gestational ages and analyzed its correlation with gestational age. Results: The peak velocities of the biventricular segments of the normal fetuses showed a decreasing trend from the basal to the middle to the apex segment, and the differences were statistically significant (P < 0.05). However, the strain and peak strain rate between adjacent segments showed no significant differences (P > 0.05). The peak global velocity of both ventricles increased with the gestational age, and it was moderately correlated with gestational age; however, the correlation of strain and peak strain rate with gestational age was not statistically significant (P > 0.05). Conclusion: In normal fetuses, the peak myocardial velocity of the biventricular segments showed a decreasing trend from the basal to the apical segment. The global peak myocardial velocity was linearly correlated with gestational age; however, the global strain and peak strain rate did not change as gestational age increased, indicating that the myocardial deformability of the fetus' ventricles was constant in the middle and late trimesters.

Pirfenidone Attenuates Renal Tubulointerstitial Fibrosis through Inhibiting miR-21.

BACKGROUND: Our previous studies had shown pirfenidone (PFD) not only improved tubulointerstitial fibrosis (TIF) but also inhibited the expression of microRNA-21 (miR-21) in the renal tissue of unilateral urethral obstruction (UUO) rats. This study aims to investigate whether PFD can attenuate TIF through inhibiting miR-21 in UUO rats. METHODS: Sprague Dawley rats were divided randomly into sham-operated group, UUO group, and PFD and olmesartan (Olm) treatment groups. Samples were collected on day 14. Expression of miR-21, TGF-ß1, Smad3, and Smad7 mRNA in the renal tissue was detected using real-time quantitative PCR. Immunohistochemistry was performed to assess the protein expressions of collagen III, E-cadherin, and α-SMA. Automated capillary Western blotting was used to detect the quantitative expression of TGF-ß1, Smad3, p-Smad3, Smad7, collagen III, E-cadherin, and α-SMA in renal tissues. The expression of miR-21 and Smad7 mRNA and the protein levels of collagen III and α-SMA were examined in the miR-21-overexpressing cell line, NRK-52E. RESULTS: Compared with the UUO group, both PFD and Olm inhibited renal tubular dilation, diffused epithelial cell degeneration and necrosis, and reduced renal interstitial edema, inflammatory cell infiltration, and collagen fiber deposition, while no significant difference between PFD group and Olm group. Informatics-based approaches identified Smad7 as a likely candidate for regulation by miR-21. Compared with the sham group, miR-21 expression was upregulated in the UUO group resulting in the downregulation of Smad7 expression due to degradation. The overexpression of miR-21 in the in vitro model downregulated Smad7 and promoted EMT and ECM accumulation. Protein levels of TGF-ß1, Smad3, p-Smad3, collagen III, and α-SMA were upregulated, while E-cadherin protein was downregulated in the UUO group than in the sham group. PFD rather than Olm decreased the expression of miR-21 and increased the expression level of Smad7 mRNA and then inhibited the TGF-ß1/Smad3 signaling pathway. Olm only downregulated the TGF-ß1/Smad3 signaling pathway. CONCLUSIONS: PFD improves TIF by downregulating the expression of miR-21, then elevating Smad7, and finally inhibiting the activation of the TGF-ß1/Smad3 signaling pathway in UUO rats.

Diagnostic efficacy and influence factors of urinary protein/creatinine ratio replacing 24-h urine protein as an evaluator of proteinuria in children.

PURPOSE: The aim was to investigate the diagnostic efficacy of urinary protein/creatinine ratio (UPCR) and factors influencing its substitutability of 24-h urine protein (24hUP) in children with proteinuria. METHODS: A total of 356 children were recruited, including 149 with non-nephrotic-range proteinuria and 207 with nephrotic-range proteinuria which were further divided into Henoch-Schönlein purpura nephritis (HSPN), lupus nephritis (LN), and primary nephrotic syndrome (PNS). The urine protein and creatinine were measured by routine methods. Bland-Altman analysis was used to test the agreement. Spearman correlation was performed to evaluate the relevance. The receiver operating characteristic curve was used to analyze the diagnostic efficacy of UPCR. RESULTS: Bland-Altman analysis showed there was an excellent agreement between UPCR and 24hUP in each group. Correlations between UPCR and 24hUP were strong in 356 children (r = 0.869) and in the non-nephrotic-range proteinuria group (r = 0.806), but moderate in nephrotic-range proteinuria group (r = 0.586). With the increase of nephrotic-range proteinuria, the correlations between UPCR and 24hUP were decreased further, however, after UPCR was adjusted by 24-h urine creatinine (24hUCr), the correlation coefficient was improved (r = 0.682). In three subgroups with nephrotic-range proteinuria, high correlation coefficient (r = 0.731) was observed in HSPN, but not in LN (r = 0.552) and PNS (r = 0.563). The sensitivity and specificity of UPCR for diagnosing nephrotic-range proteinuria were 89.9 % and 92.2%. CONCLUSIONS: UPCR is competent in evaluating proteinuria. The degree of proteinuria, 24hUCr and the underlying pathological types of renal disease may be the important influencing factors in the correlation between UPCR and 24hUP in children with nephrotic-range proteinuria.

Application of a combination of echocardiographic techniques in an experimental model of epirubicin-induced cardiotoxicity.

This study compared the potential ability of multinomial echocardiographic parameters in early detection, prediction and combined diagnosis of antineoplastic-related cardiotoxicity. Male Balb/c mice were repeatedly administered with low doses of epirubicin (6 × 3 mg/kg; n = 20) to induce cardiac injury or with placebo as control (n = 10). Conventional and strain parameters as well as myocardial performance index (MPI) were analyzed at baseline, 1 day after the second, fourth and sixth cycle, and 12 days after completion of chemotherapy (as follow-up) by a high-resolution rodent ultrasound machine. After the experiment, serum cTnI levels were measured, and myocardial injury was evaluated by histological analyses. Thirteen mice developed cardiotoxicity after epirubicin exposure. Global longitudinal (GLS), radial strain (GRS) and longitudinal strain rate (LSR) were markedly decreased (all P ≤ 0.01) and MPI was increased (P ≤ 0.05) at the completion of treatment compared with baseline values. GLS expressed the best correlations with myocardial pathological injury, especially with collagen content (ρ = - 0.68, P < 0.01). Additionally, GLS and MPI were associated with serum cTnI levels. A > 9.5% decrease in GLS from baseline to the fourth cycle of chemotherapy could predict future cardiotoxicity (odds ratio = 0.331, P < 0.05). GLS (cutoff value, - 15.16%) combined with MPI (cutoff value, 0.64) could improve the accuracy of diagnosing cardiotoxicity (sensitivity, 92%; specificity, 87%). GLS was the only predictor of cardiotoxicity. GLS combined with MPI may provide a noninvasive and accurate method for the early detection of cardiotoxicity.

MicroRNA-128/homeobox B8 axis regulates ovarian cancer cell progression.

MicroRNA-128 (miR-128) has been found to be dysregulated and might function as a tumour suppressor in various cancers, including ovarian cancer. However, the underlying mechanism of miR-128 in ovarian cancer has not been fully understood. The miR-128 and homeobox B8 (HOXB8) levels in clinical samples and cultured cell lines were measured using qRT-PCR and/or Western blot analysis. Cell proliferation was assessed using Cell Counting Kit-8 assay. Cell apoptosis was determined using flow cytometry. The association between miR-128 and HOXB8 was confirmed using dual-luciferase reporter assay. Results showed that decreased miR-128 expression and increased HOXB8 expression were observed in ovarian cancer tissues and cell lines. Transfection with miR-128 mimics suppressed the cell proliferation and enhanced paclitaxel sensitivity in ovarian cancer cell lines. miR-128 directly targeted HOXB8 in ovarian cancer cell lines. Knockdown of HOXB8 abolished the effects of miR-128 inhibitor on ovarian cancer cell proliferation and paclitaxel sensitivity. Summarily, miR-128 displayed a tumour suppressor role in ovarian cancer via targeting HOXB8. It is supposed that miR-128 might be effective for targeting therapy for ovarian cancer.

Urinary metal and metalloid biomarker study of Henoch-Schonlein purpura nephritis using inductively coupled plasma orthogonal acceleration time-of-flight mass spectrometry.

To obtain a better understanding as to whether concentration alterations of metals and metalloids in urine were related to Henoch-Schonlein purpura nephritis (HSPN), the profiles of as many as 29 elements in urine were compared among three groups, the Henoch-Schonlein purpura (HSP), HSPN and a healthy control group. To this end, a reliable method has been developed for the simultaneous quantification of multiple elements including Li, Be, B, Al, Sc, Ti, V, Cr, Mn, Fe, Ni, Co, Cu, Zn, Ga, Ge, As, Se, Rb, Sr, Mo, Cd, Sn, Sb, Cs, Ba, Tl, Pb and Bi in urine using inductively coupled plasma orthogonal acceleration time-of-flight mass spectrometry (ICP-oa-TOF-MS). The process of sample pre-treatment used a direct 20-fold dilution method with centrifuged urine. The internal standard element used for quantification was 103Rh, and 1,4-butanediol was chosen as a matrix matching reagent. The method detection limits of these 29 elements were in the range of 0.04-12ngmL-1. Results of statistical analysis revealed that the concentrations of 15 elements and the element homeostasis were significantly different among these three groups. Our study provides a potential method for HSPN metal and metalloid biomarker discovery.