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Brain iron deposition in hemodialysis (HD) patients increases over time. Iron deficiency in gray matter nuclei has been reported to lead to idiopathic restless legs syndrome (RLS) symptoms. Regardless of unpleasant RLS sensations, the patterns of iron deposition between hemodialysis patients with RLS (HD-RLS) and hemodialysis patients without RLS (HD-nRLS) are still unclear. To evaluate the differences in iron deposition patterns between HD-RLS and HD-nRLS patients, we utilized quantitative susceptibility mapping (QSM).In sum, 24 HD-RLS patients, 25 HD-nRLS patients and 30 age- and sex-matched healthy controls (HCs) were enrolled. The QSM was used to assess susceptibility values of the regions of interest (ROIs), including the caudate nucleus (CN), putamen (PUT), globus pallidus (GP), thalamus (THA), substantia nigra (SN), red nucleus (RN) and dentate nucleus (DN).HD duration was significantly longer in HD-RLS patients than in HD-nRLS patients (P 0.05). The susceptibility of HD-RLS and HD-nRLS patients in PUT was higher than that in HCs (P 0.05), illustrating elevated iron content in the nucleus. Compared with HD-nRLS patients, HD-RLS patients demonstrated reduced susceptibility in CN and PUT (both P 0.05). Compared with HCs, HD-RLS patients displayed decreased susceptibility in DN (P 0.05).Different iron deposition patterns between HD-RLS and HD-nRLS patients in PUT and DN, which further support disturbed sensory processing in RLS, may be involved in RLS pathogenesis in HD patients.

Trastuzumab resistance is one of the principal causes of failure in tumor chemotherapy. Previous studies showed that the DNA-binding activity of GATA6, a transcription factor, had a remarkable enhancement in trastuzumab resistant gastric cancer cells (NCI N87/R), while its correlation to resistance remains unclear. In this study, Crispr/Cas9 was employed to establish a GATA6 knock-out cell line (NCI N87R/GATA6). The signaling pathways regulated by GATA6 and related to trastuzumab resistance were investigated based on label-free quantitative proteomics combined with bioinformatics. The extracted proteins were alkylated, and digested using filter aided sample preparation (FASP), and the peptides were separated via high performance liquid chromatography (HPLC) and thereafter quantified by liquid chromatography coupled with tandem mass spectroscopy (LC-MS/MS). Differentially expressed proteins were screened by fold change and student's t-test between NCI N87/R and NCI N87R/GATA6 cells. WebGestalt website was adopted for Gene ontology analysis, and GeneAnalytics database was utilized for pathway enrichment analysis. The results demonstrated that GATA6 knock-out enhanced the antiproliferative effect of trastuzumab on NCI N87/R cells and suppressed their invasion ability. A total of 5792 proteins were quantified by LC-MS/MS, among which 305 proteins were up-regulated in NCI N87R/GATA6 cells while 182 ones down-regulated. Pathway enrichment analysis revealed that mitochondrial transport, apoptosis, DNA damage, glucose metabolism, pyruvate metabolism and TCA cycle, and Wnt/β-catenin degradation pathways exhibited significant changes. Western blotting manifested that the expression of mitochondrial dyneins OPA1 and DNM1L, apoptosis protein caspase-9, TCA metabolic enzymes SUCLG2 and MDH1, and glycogen metabolic enzyme PYGL changed significantly in NCI N87R/GATA6 cells, manifesting that GATA6 knock-out gave rise to mitochondrial dysfunction and abnormal energy metabolism, and therefore inducing the apoptosis of NCI N87R/GATA6 cells. These results implicated that inhibiting the transcriptional activity of GATA6 could be an effective strategy to reverse trastuzumab resistance in gastric cancer cells.

Left ventricular mass (LVM) is related to poor outcomes of HD patients. Possible contributions of vascular calcification to LVM changes over time have not been well established. This is a prospective cohort study. At baseline, left lateral lumbar spine radiograph was conducted. The echocardiogram was performed respectively at baseline and 2 years later. The dialysis records were documented in the first week of the first month for each quarter during the two years. Fasting blood samples were collected. The averages were calculated. After the second echocardiogram, patients were followed up for another five years. One hundred and four patients were enrolled, with 57 males (54.8%) and an average age of 60.0 ± 11.1 years. Ninety-one patients had AAC (87.5%). The average level of LVMI changes 2 years later was 3.31 (− 1.51 ~ 8.18) g/m2.7, and 68 patients had increased LVMI. After another five years, 28 patients died (26.9%). Patients with worsening LVMI had significantly higher baseline AAC scores (8.0 (3.0 ~ 12.0) vs 4.0 (1.3 ~ 7.0), P = 0.022). The 5-year mortality rate was also higher (23/68 vs 5/31, P = 0.029). Multiple stepwise regression analysis demonstrated that baseline AAC was an independent predictor for increased LVMI (P = 0.005). For one point increase in AAC scores, LVMI increased by 0.27g/m2.7 2 years later. The independent correlated factors baseline AAC were age and hs-CRP. In conclusion, AAC is an independent predictor of LVMI deterioration over time in HD patients. Whether direct intervention for LVM can benefit MHD patients is controversial. Prevention and treatment of VC may be an important intervention target to improve LVM and prognosis.

Objective: This study's objective is to explore the effect of venous calcium (Ca) load on Ca metabolism using an isotope tracer in both healthy rats and rats with chronic renal failure (CRF). Materials and Methods: The methods included inducing CRF by feeding rats with food that containing 0.75% adenine. Both the rats in the control and CRF group received an intravenous injection of 100 mg/kg CaCl2 mixed with 0.5 mg/kg 42Ca for over 2 minutes in order to induce hypercalcemia. Blood samples, urine, and feces were collected for Ca kinetics and metabolism testing before and 24 hours after the injection. Results: Peak blood Ca levels were presented at 5 minutes in both groups but returned to baseline levels differently: within 3 hours in the CRF group and 1 hour in the control group. 24-hour hypercalciuria and a higher weight-corrected fecal Ca loss were consistently detected in the CRF rats before and after the injection, along with a decrease in the percent fractional Ca absorption and true fractional calcium absorption compared to the control group. The increased fecal Ca in the CRF rats primarily resulted from changes in decreased absorption rate, which indirectly lead to an increase in endogenous fecal Ca (EFC). Body weightcorrected EFC levels in the CRF group were significantly higher than those in the control group (207.58 ± 107.18 mg/kg/day vs. 62.44 ± 20.49 mg/kg/day). In addition, EFC was 22.6% and 26.02% of the total fecal Ca in the CRF group and the control group. Conclusion: These results may be useful in the evaluation of calcium metabolism and may have important implications for stable isotopes that are widely used in exploring calcification studies in the CRF.

HLE-B3 cell line, a human lens epithelial cell line, was used to examine the anti-glycative and anti-oxidative protection of aqueous extract prepared from steamed red amaranth leaves against high glucose induced injury. Phytochemical profile of this aqueous extract was analyzed. HLE-B3 cells were pretreated by this aqueous extract at 0.25%, 0.5%, or 1%, and followed by high glucose treatment. Results showed that the content of phenolic acids, flavonoids, anthocyanins, carotenoids, and triterpenoids in this aqueous extract was in the range of 1,107-2,861 mg/100 g dry weight. High glucose decreased cells viability and suppressed Bcl-2 mRNA expression. This aqueous extract pretreatments raised 11-42% cell survival and upregulated 20-47% Bcl-2 mRNA expression. High glucose reduced Na+ -K+ ATPase activity and mitochondrial membrane potential (MMP). This aqueous extract raised 27-40% Na+ -K+ ATPase activity, and 18-51% MMP. High glucose stimulated the generation of total advanced glycative endproducts (AGEs), methylglyoxal, and reactive oxygen species (ROS). This aqueous extract pretreatments lowered total AGEs, methylglyoxal, and ROS levels in the range of 0.38-1.17 folds, 1.7-4.9 nmol/mg protein, and 0.35-1.06 relative fluorescence unit/mg protein. High glucose upregulated mRNA expression of aldose reductase, nuclear factor kappa B, and p38. This aqueous extract pretreatments decreased mRNA expression of these factors in the range of 75-159%, 57-151%, and 54-166%. High glucose downregulated mRNA expression of nuclear factor E2-related factor 2 (Nrf2). This aqueous extract pretreatments increased 12-38% Nrf2 mRNA expression. These results suggested that this aqueous extract might be a potent nutritional supplement to prevent diabetic retinopathy.

BACKGROUND: Hemodialysis (HD) causes various nervous system abnormalities. Alterations in white matter (WM) microstructure after long-term HD have been reported in a few previous studies; however, no studies have been performed to investigate enlarged perivascular spaces (PVS) in WM regions. We measured cerebral blood flow (CBF) and white matter volume (WMV) in HD patients to assess enlarged PVS severity in the WM across the whole brain and suggest possible explanations for this. METHODS: Fifty-one HD patients and 51 age-, sex-, and education-matched healthy controls (HCs) were recruited. The number of enlarged PVS in the centrum semiovale (CS), cerebral watershed (CW), and basal ganglia (BG) regions were assessed by T2-weighted MRI. CBF was estimated by arterial spin labeling (ASL), which is a non-invasive perfusion imaging technique. WMV was assessed by the computational anatomy toolbox (CAT12), which is a statistical analysis package. Differences in descriptive variables (two-tailed t-tests, χ(2) tests, Mann-Whitney U tests, and Friedman M tests), an intra-class correlation between radiologists, the relationship between enlarged PVS number and HD duration, normalized CBF and WMV (multiple regression), and group differences in CBF and WMV {voxel-wise t-tests with age and sex as covariates [cluster size >50 voxels, false discovery rate (FDR) corrected, P

BACKGROUND Endovascular treatment of acute thrombosed arteriovenous grafts performed completely under ultrasound guidance has rarely been reported. We compared the efficacy of a new endovascular thrombectomy technique (percutaneous manual thromboaspiration through the introducer sheath) with classical hybrid thrombectomy (minimally invasive surgical thrombectomy combined with high-pressure angioplasty) performed completely under ultrasound guidance, for arteriovenous graft thrombosis. METHODS This was a retrospective cohort study involving patients receiving hemodialysis who underwent arteriovenous graft thrombectomy between January 2014 and December 2017. We divided 130 participants into an intervention (endovascular) group (N.=65) and a control (classical hybrid) group (N.=65) according to the thrombectomy technique. The procedural success rate, immediate outcomes and patency were compared between the groups. RESULTS There was no significant difference in the procedural success rate (92.31% vs. 89.23%, P=0.55) between the intervention and control groups, respectively. No major complications were noted, but two cases of vessel rupture occurred in the control group and three cases of vessel rupture occurred in the intervention group. The procedure time in the intervention group was significantly shorter than that in the control group (74±14.21 min vs. 109.05±19.20 min, respectively; P

Hemodialysis with restless legs syndrome (HD-RLS) is associated with alterations in neuronal function, the blood-brain barrier and iron deposition, thus affecting cerebral metabolism and perfusion. This study utilized three-dimensional arterial spin labeling (ASL) to identify HD-RLS-related perfusion patterns and potential relationships with disease severity. Twenty-six HD-RLS patients, 30 hemodialysis patients without restless legs syndrome (HD-nRLS) and 30 age-, sex-, and education-matched healthy controls were included in this study. One-way analysis of covariance and post hoc analyses were used to assess differences in cerebral blood flow (CBF) values, demographics and clinical data among the three groups. Pearson’s correlation analysis was conducted between altered CBF values in the HD-RLS group and clinical data. Compared with HD-nRLS patients, HD-RLS patients showed increased CBF in the right primary motor cortex (false discovery rate [FDR]-corrected P

PURPOSE The purpose of this study was to investigate the differences of gray matter volume (GMV) alteration patterns between hemodialysis with restless legs syndrome (HD-RLS) and hemodialysis without restless legs syndrome (HD-nRLS) patients using voxel-based morphometry. METHODS Twenty-three HD-RLS patients, 27 HD-nRLS patients, and 27 age-, sex-, and education-matched healthy controls were included in this study. One-way analysis of covariance and post hoc analyses were used to assess differences in GMV, demographics, and clinical data among the 3 groups. Pearson correlation analysis was conducted between altered GMV in the HD-RLS group and clinical data. RESULTS Compared with HD-nRLS patients, HD-RLS patients showed decreased GMV in the left primary motor cortex (false discovery rate corrected, P < 0.05). Compared with the healthy controls, both HD subgroups (ie, those with and without RLS) exhibited consistent GMV changes, including decreased GMV in the bilateral anterior cingulate and paracingulate gyrus and left middle temporal gyrus (false discovery rate corrected, P < 0.05). The GMV values in the left precentral gyrus were negatively correlated with the RLS rating scores (r = 0.2138, P = 0.0263). CONCLUSIONS This abnormal decreased GMV in the sensorimotor cortex provides evidence for a sensory processing disorder in RLS that may be involved in the pathogenesis of RLS in HD patients.

Recent studies have demonstrated that hemodialysis patients exhibit disruptions in functional networks with invisible cerebral alterations. We explored the alterations of functional connectivity in hemodialysis patients using the graph-theory method. A total of 46 hemodialysis patients (53.11 ± 1.58 years, 28 males) and 47 healthy controls (55.57 ± 0.86 years, 22 males) were scanned by using resting-state functional magnetic resonance imaging. The brains of these patients were divided into 90 regions and functional connectivity was constructed with the automatic anatomical labeling atlas. In the defined threshold range, the graph-theory analysis was performed to compare the topological properties including global, regional and edge parameters between the hemodialysis and the healthy control groups. Both hemodialysis patients and healthy control subjects demonstrated common small-world property of the brain functional connections. At the global level, the parameters normalized clustering coefficients and small-worldness were significantly decreased in hemodialysis patients compared with those noted in healthy controls. At the regional level, abnormal nodal metrics (increased or decreased nodal degree, betweenness centrality and efficiency) were widely found in hemodialysis patients compared with those of healthy controls. The network-based statistical method was employed and two disrupted neural circuits with 18 nodes and 19 edges (P = 0.0139, corrected) and 10 nodes and 11 edges (P = 0.0399, corrected) were detected. Of note, the edge-increased functional connectivity was associated with the salience network and the frontal-temporal-basal ganglia connection, whereas the edge-decreased functional connectivity was associated with the frontoparietal network. The graph-theory method may be one of the potential tools to detect disruptions of cerebral functional connectivity and provide important evidence for understanding the neuropathology of hemodialysis patients from the disrupted network organization perspective.

Structural and functional brain alterations have been always observed in end-stage renal disease (ESRD) patients undergoing hemodialysis. The present study aimed to investigate the gray matter volume (GMV) changes in hemodialysis patients compared with those noted in healthy subjects, as well as explore the associated functional connectivity alterations based on the abnormal GMV regions. The experiments revealed the effects of regional morphometry aberrance on the brain functional integrity. A total of 46 hemodialysis patients (53.11 ± 1.58 years, 28 males) and 47 healthy subjects (55.57 ± 0.86 years, 22 males) were enrolled in the present study. All subjects underwent high-resolution T1-weighted imaging, resting-state functional MR imaging, and laboratory examinations were performed in hemodialysis patients. The GMV deficits were analyzed using voxel-based morphometry (VBM) and regions with GMV alteration were defined as seeds for functional connectivity analysis. Correlation analyses between significantly different regions and the results of the blood examination were further performed. We found that bilateral thalamus exhibited significantly increased volumes in the hemodialysis patients compared with those of the healthy subjects. However, the bilateral rectus, bilateral caudate, and bilateral temporal gyrus demonstrated significantly decreased volumes. When the regions with GMV alterations were defined as seeds, the hemodialysis patients exhibited decreased integrations in the thalamo-cortical network and within the basal-ganglia connection. The present study revealed the presence of different types of structural and functional brain impairments in hemodialysis patients.

Gynura bicolor leaf aqueous extract (GAE) is rich in phytochemicals including phenolic acids, flavonoids, carotenoids, and anthocyanins. Effects of GAE upon hepatic injury in mice with chronic ethanol intake were examined. Lieber-DeCarli liquid diet with ethanol was used to induce hepatic lipid accumulation, oxidative, glycative, and inflammatory injury. GAE at 0.25% or 0.5% was added in feeds, and supplied to mice consumed Lieber-DeCarli liquid diet with ethanol for 6 wk. Blood and liver were collected for analyses. Results showed that ethanol increased plasma and hepatic triglyceride and cholesterol content, and affected plasma levels of insulin, adiponectin, leptin, and ghrelin. GAE at both doses decreased lipid accumulation, and at high dose improved hormones abnormality. Histological data revealed that GAE supplement mitigated hepatic lipid deposit. Ethanol increased plasma Ne -(carboxyethymethyl)-lysine and pentosidine levels. GAE at high doses lowered those glycative factors. Ethanol depleted glutathione content, increased CYP2E1 activity and reactive oxygen species production, and reduced the activity of glutathione peroxide, glutathione reductase and catalase in liver. GAE supplement at both doses reversed these alterations and attenuated hepatic oxidative stress. GAE supplement also at both doses decreased hepatic inflammatory cytokines release in ethanol treated mice. These findings support that leaves of G. bicolor is a functional food with liver protective activities against ethanol.

Background Altered cerebral blood flow (CBF) and amplitude of low-frequency fluctuation (ALFF) have been reported in hemodialysis patients. However, neurovascular coupling impairments, which provide a novel insight into the human brain, have not been reported in hemodialysis patients. Methods We combined arterial spin labeling (ASL) and blood oxygen level dependent (BOLD) techniques to investigate neurovascular coupling alterations and its relationships with demographic and clinical data in 46 hemodialysis patients and 47 healthy controls. To explore regional neuronal activity, ALFF was obtained from resting-state functional MRI. To measure cerebral vascular response, CBF was calculated from ASL. The across-voxel CBF–ALFF correlations for global neurovascular coupling and CBF/ALFF ratio for regional neurovascular coupling were compared between hemodialysis patients and healthy controls. Two-sample t-tests were used to compare the intergroup differences in CBF and ALFF. Multiple comparisons were corrected using a voxel-wise false discovery rate (FDR) method (P < 0.05). Results All hemodialysis patients and healthy controls showed significant across-voxel correlations between CBF and ALFF. Hemodialysis patients showed a significantly reduced global CBF–ALFF coupling (P = 0.0011) compared to healthy controls at the voxel-level. Of note, decreased CBF/ALFF ratio was exclusively located in the bilateral amygdala involved in emotional regulation and cognitive processing in hemodialysis patients. In hemodialysis patients, the decreased CBF (right olfactory cortex, anterior cingulate gyrus and bilateral insula) and ALFF (bilateral precuneus and superior frontal gyrus) were mainly located in the default mode network and salience network-related regions as well as increased CBF in the bilateral thalamus. Conclusions These novel findings reveal that disrupted neurovascular coupling may be a potential neural mechanism in hemodialysis patients.

Resistance to trastuzumab, which specifically target HER2-positive breast and gastric cancer, can develop ultimately in cancer patients. However, the underlying mechanisms of resistance in gastric cancer have not been fully elucidated. Here, we established trastuzumab-resistant MKN45 and NCI N87 gastric cancer sublines from their parental cells. The resistant cells exhibited characteristics of epithelial-mesenchymal transition (EMT) and acquired higher migratory and invasive capacities. To exploit the activated pathways and develop new strategies to overcome trastuzumab resistance, we investigated MKN45 and MKN45/R cells via label-free quantitative proteomics, and found pathways that were altered significantly in MKN45/R cells, with the Wnt/&beta, catenin pathway being the most significant. We further confirmed the activation of this pathway by detecting its key molecules in MKN45/R and NCI N87/R cells via Western blot, in which Wnt3A, FZD6, and CTNNB1 increased, whereas GSK-3&beta, decreased, manifesting the activation of the Wnt/&beta, catenin pathway. Correspondingly, inhibition of Wnt/&beta, catenin pathway by ICG-001, a specific Wnt/&beta, catenin inhibitor, preferentially reduced proliferation and invasion of trastuzumab-resistant cells and reversed EMT. Concurringly, CTNNB1 knockdown in stable cell lines potently sensitized cells to trastuzumab and induced more apoptosis. Taken together, our study demonstrates that the Wnt/&beta, catenin pathway mediates trastuzumab resistance, and the combination of Wnt/&beta, catenin inhibitors with trastuzumab may be an effective treatment option.

The apoptotic effects of maslinic acid (MA) at 4, 8, 16, 32 and 64 μmol/L on human lung cancer A549 cells under normoxic and hypoxic conditions were examined. MA at 4–64 and 16–64 μmol/L lowered Bcl-2 expression under normoxic and hypoxic conditions, respectively (p &lt, 0.05). This agent at 4–64 μmol/L decreased Na+-K+-ATPase activity and increased caspase-3 expression under normoxic conditions, but at 8–64 μmol/L it caused these changes under hypoxic conditions (p &lt, 0.05). MA up-regulated caspase-8, cytochrome c and apoptosis-inducing factor expression under normoxic and hypoxic conditions at 8–64 μmol/L and 32–64 μmol/L, respectively (p &lt, 0.05). MA down-regulated hypoxia-inducible factor (HIF)-1α, vascular endothelial growth factor (VEGF), survivin and inducible nitric oxide synthase (iNOS) expression under normoxic and hypoxic conditions at 8–64 and 16–64 μmol/L, respectively (p &lt, 0.05). After cells were pre-treated with YC-1, an inhibitor of HIF-1α, MA failed to affect the protein expression of HIF-1α, VEGF, survivin and iNOS (p &gt, 0.05). MA at 8-64 and 32-64 μmol/L reduced reactive oxygen species and nitric oxide levels under both conditions (p &lt, 0.05). These findings suggest that maslinic acid, a pentacyclic triterpenic acid, exerted its cytotoxic activities toward A549 cells by mediating mitochondrial apoptosis and the HIF-1α pathway.

Epithelial-mesenchymal transition (EMT) of tubular epithelial cells is a key event in renal interstitial fibrosis and the progression of chronic kidney disease (CKD). Apelin is a regulatory peptide involved in the regulation of normal renal hemodynamics and tubular functions, but its role in renal fibrosis remains unknown. In this study, we examined the inhibitory effects of apelin on transforming growth factor-β1 (TGF-β1)-induced EMT in HK-2 cells, and evaluated its therapeutic efficacy in mice with complete unilateral ureteral obstruction (UUO). In vitro, apelin inhibited TGF-β1-mediated upregulation of α-smooth muscle actin (α-SMA) and downregulation of E-cadherin. Increased levels of phosphorylated Smad-2/3 and decreased levels of Smad7 in TGF-β1-stimulated cells were reversed by apelin co-treatment. In the UUO model, administration of apelin significantly attenuated renal interstitial fibrosis, as evidenced by the maintenance of E-cadherin and laminin expression, and markedly suppressed expression of α-SMA, TGF-β1 and its type I receptor, as well as interstitial matrix components. Interestingly, in UUO mice, there was a reduction in the plasma level of apelin, which was compensated by upregulation of APJ expression in the injured kidney. Exogenous supplementation of apelin normalized the level of plasmatic apelin and renal APJ. In conclusion, our study provides the first evidence that apelin is able to ameliorate renal interstitial fibrosis by suppression of tubular EMT through a Smad-dependent mechanism. The apelinergic system itself may promote some compensatory response in the renal fibrotic process. These results suggest that apelin has potential renoprotective effects and may be an effective agent for retarding CKD progression.

Hypertension is an independent risk factor for mortality in chronic kidney disease (CKD) and is suboptimally controlled worldwide. Therefore, this study aimed to examine the rate of BP control and the main barriers to achieving target BP, according to K/DOQI guidelines, in China. We performed a single-center, prospective cohort study. Two hundred and sixty CKD patients were referred by general physicians to nephrologists, and their BP was treated in accordance with K/DOQI guidelines for a 1-year follow-up. We evaluated improvement of BP target achievement and factors affecting BP control. We defined “not-at-goal” as persistence of systolic BP ≥ 130 mmHg and/or diastolic BP ≥ 80 mmHg after 1 year. The BP decreased from 138 ± 12/84 ± 7 mmHg at baseline to 124 ± 13/73 ± 7 mmHg after 1 year. The rate of achieving the BP goal (

Peritoneal transport status is important not only for prescription, but also as a prognostic index. Flt-1 and Flk-1, the major vascular endothelial growth factor receptors involved in angiogenesis and hyperpermeability, may play a potent role in determining peritoneal transport characteristics. However, the relationship between them has not been studied to date. We hypothesized that Flt-1 and Flk-1 expression in the peritoneal vasculature of uremic patients could be closely related to baseline peritoneal transport status.Thirty-six new patients without a previous history of peritonitis were enrolled. Clinical parameters such as age, sex, height, weight, causes of renal failure, and residual renal function were assessed. Parietal peritoneal biopsies were obtained during implantation of peritoneal dialytic catheters. Flt-1 and Flk-1 were semi-quantitatively evaluated by immunohistochemical staining. Peritoneal microvascular density (MVD) was counted. Within 6 weeks after commencing peritoneal dialysis, a standard peritoneal equilibration test was performed, and the dialysate-to-plasma concentration ratio for creatinine at 4 h (D4/P Cr) was determined. The patients were divided into two groups based on the D4/P Cr: more than 0.65 (Group H, n = 22) and less than or equal to 0.65 (Group L, n = 14). The 24-h peritoneal protein excretion (PPE) was assayed. Flt-1 and Flk-1 were correlated with peritoneal MVD, D4/P Cr, and PPE.Flt-1 and Flk-1 were detected in the peritoneal vasculature of uremic patients. Flt-1 expression was similar between the two groups, but Flk-1 expression in Group H was significantly higher than that in Group L (p = 0.001). Flt-1 expression did not show significant correlations with peritoneal MVD, D4/P Cr, and PPE. However, Flk-1 expression showed significant correlations with the above three parameters (p0.001 for all).For the first time, the expressions of Flt-1 and Flk-1 in peritoneal vasculature of uremic patients were detected. Flk-1 expression in peritoneal vasculature of uremic patients is closely correlated with the number of peritoneal microvessels, peritoneal small solute transport rate, and PPE. Our findings strongly suggest that Flk-1 may be a crucial determinant of baseline peritoneal transport characteristics. Further interventional studies are needed.

To analyze predictive factors for all-cause mortality, cardiovascular (CV) mortality, nonfatal CV events (CVE) in maintenance hemodialysis (MHD) patients, and to compare the effects of standard hemodialysis (HD) and online hemodiafiltration (HDF) on these factors and outcomes. A total of 333 MHD patients were prospectively followed up for 50 ± 15 months and all-cause death, CV death and CVE were registered. At the baseline, demographic, clinical, and laboratory data of the whole population were recorded. Then, patients were stratified into two groups according to the dialysis modalities, HD (n = 268) and HDF (n = 65). At the end of 6th month, clinical and laboratory data were recorded again. The predictive factors at baseline for all-cause mortality, CV mortality, and CVE were analyzed by Cox regression. The effects of HD and HDF on these factors at the 6th month and long-term outcomes were compared by t-test and Kaplan-Meier method, respectively. Age, gender, left ventricular mass index (LVMI), aortic arch calcification score (AoACS), hemoglobin (Hb)10 g/dL, and ferritin500 ng/mL maintained independent associations with all-cause mortality. C-reactive protein (CRP), LVMI, AoACS, and Hb10 g/dL were associated with CV mortality. Prior cardiovascular disease (CVD), AoACS and LVMI were independent predictors of nonfatal CVE. Higher body mass index (BMI), body weight, total serum cholesterol, Hb concentration, and lower CRP level, LVMI, and AoACS were found in patients on HDF at the end of the 6th month. Improved outcomes with longer survival time for all-cause mortality, CV mortality, and CVE were found in HDF group. Age, gender, LVMI, AoACS, Hb, and ferritin were predictors of all-cause mortality in MHD patients. CRP, LVMI, AoACS, and Hb were associated with CV mortality. Prior CVD, AoACS, and LVMI were independent predictors of nonfatal CVE. HDF could improve BMI, body weight, total serum cholesterol, Hb, CRP, LVMI, AoACS, and long-term outcomes, including all-cause mortality, CV mortality, and CVE.

Asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase inhibitor, is associated with vascular dysfunction. The polypeptide apelin mediates two major actions on blood vessels. However, their combined effects on vascular function are not fully understood. The present study aimed to determine the effect of apelin-13 on myosin light chain (MLC) phosphorylation in vascular smooth muscle cells (VSMCs) under ADMA-induced endothelial leakage conditions. To assess the increased permeability induced by ADMA, human umbilical vein endothelium cells (HUVECs) were plated in transwell dishes. The FITC-dextran flux and FITC-apelin-13 flux through the endothelial monolayer were measured. To examine the effect of leakage of apelin-13 on MLC phosphorylation in HUVSMCs, transwell dishes were used to establish a coculture system with HUVECs in upper chambers and HUVSMCs in lower chambers. Western blot was performed to assess the phospho-MLC levels. ADMA increased endothelial permeability in a concentration- and time-dependent manner, accompanied by actin stress fiber assembly and intercellular gap formation. When HUVECs were treated with ADMA, the permeability to both macromolecular dextran and micromolecular apelin-13 increased significantly. Both p38 MAPK inhibitor and NADPH oxidase inhibitor could prevent HUVECs from the increased permeability, and the changes of cytoskeleton and intercellular junction, which were induced by ADMA. Apelin-13 passed through the ADMA-stimulated endothelial monolayer and increased the expression of phospho-MLC in VSMCs. These results suggest that ADMA increases endothelial permeability, which may involve the p38 MAPK and NADPH oxidase pathway. Apelin-13 can pass through the damaged endothelial barrier, and acts directly on VSMCs to increase MLC phosphorylation.

Common variable immunodeficiency disease (CVID) represents a heterogeneous group of primary hypogammaglobulinemias of unknown etiology, characterized by decreased serum immunoglobulin levels and recurrent bacterial infections and is often accompanied by autoimmune disease. Renal involvement is rare in CVID, despite widespread involvement of other organ systems. We describe a 21-year-old girl who presented with recurrent infections, hepatosplenomegaly, renomegaly, and renal insufficiency. Renal biopsy revealed a remarkable diffused interstitial infiltration and severe degenerative tubular lesions. Interstitial infiltration consisted mainly of CD8(+) T cells and CD68(+) macrophages with less CD4(+) T and rare B cells. For the cases with recurrent infections, multiple organomegaly, and renal insufficiency, clinicians should consider to exclude CVID, so as to make the timely diagnosis and appropriate management.

Mice treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) were used to examine the neuroprotective effects of carnosine. Carnosine at 0.5, 1, and 2 g/L was directly added to the drinking water for 4 weeks. MPTP treatment significantly depleted striatal glutathione content, reduced the activity of glutathione peroxidase (GPX), superoxide dismutase (SOD), and catalase, increased malondialdehyde and reactive oxygen species levels, and elevated interleukin-6, nitrite, and tumor necrosis factor-α production as well as enhanced inducible nitric oxide synthase (iNOS) activity in the striatum (P < 0.05). The preintake of carnosine significantly attenuated MPTP-induced glutathione loss, retained the activity of GPX and SOD, diminished oxidative stress, and lowered inflammatory cytokines and nitrite levels as well as suppressed iNOS activity (P < 0.05). MPTP treatment significantly suppressed GPX mRNA expression and enhanced iNOS mRNA expression (P < 0.05). Carnosine preintake significantly elevated GPX mRNA expression and declined iNOS mRNA expression (P < 0.05). Preintake of carnosine also significantly improved MPTP-induced dopamine depletion and maintained 3,4-dihydroxyphenylacetic acid and homovanillic acid levels (P < 0.05). These results suggest that carnosine could provide antioxidative and anti-inflammatory protection for the striatum against the development of Parkinson's disease.

Asymmetric dimethylarginine (ADMA) as a uremia toxin is accumulated in end-stage renal disease (ESRD) patients. Elevated ADMA level has been shown to be predictive of cardiovascular diseases (CVDs) and all-cause mortality in ESRD. Therefore, we investigated the removal of ADMA by different dialysis treatments.There were 30 each of hemodialysis (HD), hemodiafiltration (HDF), peritoneal dialysis (PD) patients, and healthy volunteers enrolled. The ADMA concentrations in serum, urine, and spent dialysate samples were determined. The urine and spent dialysate volumes were recorded. The ADMA removals by urine and spent dialysate in 1 week were calculated and compared among four groups. It was also analyzed for the correlations between the total removal of ADMA in 1 week and the parameters of age, durance of dialysis, glomerular filtration rate, urine volume, urinal ADMA level, spent dialysate volume, and spent dialysate ADMA level.The serum levels of ADMA in dialysis patients were much higher than in healthy subjects (0.32 +/- 0.09 micromol/L), and their 1-week total removals of ADMA were much lower than healthy controls (249.21 +/- 57.04 micromol/week) (p-values all were less than 0.01). Among dialysis groups, serum ADMA levels decreased significantly in PD patients compared with HD or HDF patients (1.38 +/- 0.30 micromol/L vs. 1.82 +/- 0.38 micromol/L and 1.63 +/- 0.32 micromol/L, p0.01), and the total removal of ADMA diminished remarkably by turns of PD, HDF, and HD groups (47.79 +/- 8.20 micromol/week, 31.79 +/- 8.92 micromol/week, 14.63 +/- 6.53 micromol/week, respectively, p0.01). The total removal of ADMA in 1 week was related directly with the spent dialysate concentrations of ADMA, the spent dialysate volume, and the urine volume.ADMA was mainly removed by dialysate in dialysis patients. Different dialysis models have different clearance capability on plasma ADMA. PD might be more effective on ADMA removal than HD and HDF, with HDF being more effective than HD.

The inhibitory effect of diallyl sulphide (DAS) and diallyl disulphide (DADS) against meticillin-resistant Staphylococcus aureus (MRSA) infection in diabetic mice was studied. The influence of these agents on the plasma levels of fibronectin, C-reactive protein (CRP), fibrinogen, interleukin (IL)-6 and tumour necrosis factor-α (TNF-α), and on the activity of plasminogen activator inhibitor-1 (PAI-1), antithrombin III (AT-III) and protein C, in MRSA-infected diabetic mice was examined. To induce diabetes, mice were treated intraperitoneally with streptozotocin for 5 consecutive days. Ten clinical MRSA isolates obtained from infected patients were used in this study. Diabetic mice were infected by injecting 200 μl MRSA/PBS suspension containing 107 c.f.u. via the tail vein. At day 4 post-infection, 200 μl DAS or DADS was administrated twice orally with an interval of 12 h. Eight hours after each administration, the blood and organs of mice were collected. Results showed that DAS and DADS significantly decreased MRSA viability in the kidney (P P α (P α (P >0.05); however, DAS or DADS given twice significantly decreased the plasma levels of both IL-6 and TNF-α (P P P >0.05), but DAS or DADS given twice significantly increased AT-III activity (P P P P

The in vitro inhibitory effects of diallyl disulphide (DADS), diallyl trisulphide (DAT), roselle calyx extract and protocatechuic acid (PA) on the growth of Helicobacter pylori (15 susceptible, 11 clarithromycin-resistant and 9 metronidazole-resistant strains) were studied. The inhibition zone was determined after each agent had been heated at 25, 60, 100 °C for 60 min. The minimal inhibitory concentration (MIC) of each agent was determined by the tube dilution assay. The results showed that heat treatment did not affect the anti-H. pylori activity of DADS, DAT, roselle calyx extract and PA, and the MIC values of these agents against test H. pylori strains were in the range 8–64 mg/L. The time-kill study assay for DAT and PA at 1× MIC was monitored in Muller Hinton broth supplemented with 10% horse blood or mice stomach homogenate. Both DAT and PA inhibited the growth of all test H. pylori in broth and mice stomach homogenate (p < 0.05); however, the inhibitory effects of these two agents were less in mice stomach homogenate than in broth (p < 0.05). DAT at 4, 6, 8, 10, 12, 14 mg/L and PA at 8, 16, 24, 32, 40, 48 mg/L were used for urease activity assay. These two agents significantly reduced urease activity of test H. pylori strains (p < 0.05), in which DAT and PA at 1× MIC reduced the urease activity of H. pylori to 70% and 40%, respectively. These agents, based on their lower MIC values and heat tolerance, might be useful in the prevention or therapy of H. pylori. Copyright © 2007 John Wiley & Sons, Ltd.

In this study, the antibacterial effects and action modes of asiatic acid against the foodborne bacterial pathogens Escherichia coli O157:H7, Salmonella Typhimurium DT104, Pseudomonas aeruginosa, Listeria monocytogenes, Staphylococcus aureus, Enterococcus faecalis, and Bacillus cereus were examined. Minimal inhibitory concentrations (MICs) of asiatic acid against these bacteria were in the range of 20-40 μg/mL. Minimum bactericidal concentrations of asiatic acid were in the range of 32-52 μg/mL. Asiatic acid at 2X MIC effectively reduced bacterial numbers from 6 log10 to < 2 log10 in all test bacteria within 6 h (P < 0.05). The antibacterial activity of asiatic acid was not affected by heat treatments from 25 to 100°C. Asiatic acid at 1 or 2X MICs caused 40-56% and 71-89% membrane damage in test bacteria within 2 h, respectively In addition, asiatic acid at 1 or 2X MICs led to 1.5-2.4 ppm and 2.9-4.1 ppm K(+) release within 2 hr, respectively. Asiatic acid treatments dose-dependently increased bacterial nucleotide leakage (P < 0.05). After 3 days of storage at 25°C, the addition of asiatic acid dose-dependently inhibited the growth of test bacteria in ground beef (P < 0.05), in which 8 mg asiatic acid treatments led to bacterial levels (log CFU/g) in said ground beef lower than 2. These findings suggest that asiatic acid might be a potent antibacterial agent to prevent food contamination.

Background: Purple laver ((Porphyra dentate) is a popular edible seaweed in Asia. This study examined protective effects of extract from purple laver extract (PLE) in diabetic mice. Methods: Content of carotenoids and anthocyanins in PLE was analyzed. PLE at 0.5 and 1% was supplied for 7 weeks. Results: PLE was rich in anthocyanins. PLE intake at 0.5 and 1% lowered plasma glucose level (P

Scope Antiglycative effects of glycyrrhizic acid (GA) in kidney of diabetic mice were examined. Methods and results GA at 0.05, 0.1, and 0.2% was supplied to diabetic mice for 9 wk. Results showed that GA intake increased its deposit in kidney, raised plasma insulin level, decreased plasma glucose and blood urine nitrogen levels, and improved creatinine clearance rate (p < 0.05). GA intake dose-dependently reduced renal carboxymethyllysine level, and at 0.1 and 0.2% decreased plasma HbA1c, urinary glycated albumin, and renal pentosidine levels (p < 0.05). Dietary GA intake declined renal aldose reductase activity and protein expression, as well as lowered renal fructose and sorbitol levels (p < 0.05). GA intake dose-dependently increased glyoxalase-1 activity and expression, and decreased renal methylglyoxal level (p < 0.05). This compound at 0.1 and 0.2% raised glyoxalase-2 activity and protein expression, and increased d-lactate formation (p < 0.05). GA intake dose-dependently suppressed renal expression of nuclear factor kappa B (NF-κB) p65 and p-p38, decreased reactive oxygen species production, and retained glutathione content (p < 0.05). This compound at 0.1 and 0.2% downregulated renal expression of NF-κB p50 and p-ERK1/2 (p < 0.05), and lowered renal level of monocyte chemoattractant protein-1 (MCP-1) and intercellular adhesion molecule-1 (ICAM-1). Conclusions These findings suggest that glycyrrhizic acid is an antiglycative and renal-protective agent.

Asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase inhibitor, is considered to be an independent risk factor in the progression of chronic kidney diseases (CKD). It can induce kidney fibrosis by increasing transforming growth factor (TGF)-β1 expression, but its molecular mechanism is unclear. The aim of the present study was to investigate the role of actin cytoskeleton in ADMA-induced TGF-β1 high expression in human renal glomerular endothelial cells (HRGECs). The structure of stress fibers was visualized by immunofluorescence, nuclear factor-κB (NF-κB) DNA-binding activity was assessed by an electrophoretic mobility shift assay and TGF-β1 expression was assessed by western blot analysis. Results showed that ADMA induced the assembly of stress fibers, DNA binding of NF-κB, and increasing expression of TGF-β1. When the dynamics of actin cytoskeleton was perturbed by the actin-depolymerizing agent cytochalasin D and the actin-stabilizing agent jasplakinolide, or ablation of stress fiber bundles by the nicotineamide adenine dinucleotide phosphate oxidase inhibitor apocynin and p38 mitogen-activated protein kinase inhibitor SB203580, ADMA-induced DNA binding of NF-κB and TGF-β1 expression were inhibited. These results revealed an actin cytoskeleton-dependent mechanism in ADMA-induced NF-κB activation and TGF-β1 high expression in HRGECs. The specific targeting of the actin cytoskeleton may be a useful strategy to prevent ADMA-activated kidney fibrosis in CKD.

The effects of trans fatty acids, elaidic acid (trans-9, C18:1) and linoelaidic acid (trans-9, trans-12 C18:2), at 20 or 40 μM in nerve growth factor differentiated PC12 cells with or without beta-amyloid peptide (Aβ) were examined. Elaidic acid treatment alone did not affect cell viability and oxidative injury associated markers (P > 0.05). However, co-treatments of elaidic acid and Aβ led to more reduction in mitochondrial membrane potential (MMP) and Na⁺-K⁺-ATPase activity, and more increase in DNA fragmentation and 8-hydroxydeoxyguanosine (8-OHdG) production than Aβ treatment alone (P < 0.05). Linoelaidic acid alone exhibited apoptotic and oxidative effects in cells via decreasing MMP and Na⁺-K⁺-ATPase activity, increasing reactive oxygen species (ROS) level, lowering glutathione content and glutathione peroxidase (GPX) activity (P < 0.05). The co-treatments of linoelaidic acid with Aβ further enhanced oxidative damage via enhancing the generation of ROS, nitrite oxide and 8-OHdG, elevating caspase-3, caspase-8 and nitric oxide synthase activities, as well as declining GPX, catalase and superoxide dismutase activities (P < 0.05). These results suggested that the interaction of linoelaidic acid and Aβ promoted oxidative stress and impaired mitochondrial functions in neuronal cells.

Residual renal function (RRF) is of paramount importance for hemodialysis (HD) adequacy, morbidity, and mortality. Some studies have shown that overhydration is beneficial for preservation of RRF, but it can also increase the probability of adverse events such as hypertension and heart failure in HD patients. To determine the optimal amount of dehydration, we performed HD with limited water removal in HD patients. Eighteen HD patients included in this self-controlled study underwent HD with limited water removal. Water removal volume was determined by a previous volume as follows. Total water removal volume was divided into levels: ≤3.0, 3.0-9.0, and9.0 L per week. Water removal was performed to obtain dry weight in the last dialysis, and was performed three times with a ratio of 1:1:2 and 2:2:3, respectively. Urine volume, endogenous creatinine clearance rate, Kt/V, hemoglobin, and serum albumin were recorded before and after the study at 3, 6, 9, and 12 months. The follow-up period was 12 months. Ten patients withdrew from the study because of adverse events including hypertension (n = 3), heart failure (n = 3), angina (n = 1), polycystic kidney rupture (n = 1), obvious edema (n = 1), and one patient had too much interdialytic weight gain to continue. As a result, we stopped this study after 1 month. Our data suggest that the preservation of RRF with limited water removal in HD patients must be interpreted with caution.

Following the introduction of modified cellulosic and then synthetic membrane dialyzers, it was realized that the dialyzer bio-incompatibility depends on the membrane composition. We designed a prospective, randomized, cohort study of 6 months to determine several parameters of biocompatibility in maintenance hemodialysis (MHD) patients treated with four different membrane dialyzers.There were 60 MHD patients enrolled in the study. In baseline, synthetic low-flux dialyzer, polysulfone (PS) membrane was used in all patients for at least 3 months. Then the patients were randomly divided into three groups according to different dialyzer membranes. Synthetic high-flux dialyzer group, polyethersulfone membrane, cellulose triacetate (CTA) high-flux membrane, and synthetic low-flux dialyzer, polymethylmethacrylate (PMMA) membrane were used in 6 months. A new dialyzer was used for each study treatment, and there was no dialyzer reuse. The biocompatibility markers and solutes removal markers were detected repeatedly at different time points.The blood levels of highly sensitive C reactive protein, interleukin (IL)-1β, and interleukin (IL)-13 showed no difference among different groups at al time points. However, the blood complement levels and white blood cell counts were significantly different among three groups. When the dialyzers changed from PS to PMMA membrane, C3a levels and white blood cell counts changed significantly (p0.05). Moreover, the changes of C5a levels were significantly different between group CTA and group PMMA in month 3 (p0.05).There were much more differences on bio-incompatibility among different dialyzer membranes.

The effects of protocatechuic acid (PCA) on hepatic activity and/or mRNA expression of lipogenic enzymes and sterol regulatory element-binding proteins (SREBPs) in mice fed a trans fatty acid (TFA)-rich diet were examined. PCA at 1, 2, or 4% was provided for 10 weeks. Results showed that TFA diet significantly enhanced hepatic activity and mRNA expression of fatty acid synthase, 3-hydroxy-3-methylglutaryl coenzyme A reductase, stearoyl-CoA desaturase-1, and SREBP-1c (P

The impact of hepatitis B virus (HBV) or hepatitis C virus (HCV) infection upon B vitamins status and antioxidative defense in infected patients was examined. Dietary record and blood levels of B vitamins and oxidative stress–associated biomarkers were determined for 195 healthy controls, 132 HBV, and 114 HCV patients. HBV-infected patients had significantly higher levels of total cholesterol, free fatty acids (FFA), and lower ghrelin level (p

Alleviative effects of s-allyl cysteine (SAC) and s-ethyl cysteine (SEC) upon methionine and choline deficient (MCD) diet-induced hepatotoxicity in mice were examined. SAC or SEC at 1g/L was added into drinking water for 7 weeks with MCD diet. MCD feeding significantly increased hepatic triglyceride and cholesterol levels, and elevated the activity of glucose-6-phosphate dehydrogenase (G6PDH), malic enzyme, fatty acid synthase (FAS) and 3-hydroxy-3-methylglutaryl coenzyme A reductase (P < 0.05). However, the intake of SAC or SEC significantly decreased hepatic triglyceride accumulation, and reduced G6PDH and FAS activities (P < 0.05). MCD feeding significantly lowered serum and hepatic glutathione (GSH) levels, increased malondialdehyde (MDA) and oxidized glutathione (GSSG) formation, and suppressed the activity and mRNA expression of glutathione peroxidase (GPX), superoxide dismutase (SOD) and catalase (P < 0.05). The intake of SAC or SEC significantly increased serum and hepatic GSH levels, decreased MDA and GSSG formation, restored the activity and mRNA expression of GPX, SOD and catalase (P < 0.05). MCD feeding significantly enhanced the mRNA expression of interleukin (IL)-1beta, IL-6, tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta1, matrix metalloproteinases-9 (MMP-9) and collagen-alpha1 (P < 0.05). The intake of SAC and SEC significantly blunted the mRNA expression of IL-1beta, IL-6, TNF-alpha, TGF-beta1 and collagen-alpha1 (P < 0.05). SEC was greater than SAC in suppressing IL-6 and TNF-alpha expression (P < 0.05), but SAC was greater than SEC in suppressing collagen-alpha1 and TGF-beta1 expression (P < 0.05). These data suggest that SAC and SEC are potent agents against MCD-induced hepatotoxicity.

In this study, we examined oxidative stress and B vitamins status in non-small cell lung cancer (NSCLC) patients at different stages. NSCLC patients were divided into 2 groups, stage III (IIIA + IIIB, n = 27) and stage IV (n = 23). A total of 16 healthy control subjects were included for comparison. Plasma levels of alpha-tocopherol, beta-carotene, vitamin C, Se, Cu, Zn, reduced glutathione (GSH), oxidized glutathione (GSSG), lipid oxidation and the activities of glutathione peroxidase (GPX), superoxide dismutase (SOD), catalase, and xanthine oxidase (XO) were determined for evaluating oxidative status in these subjects. B vitamins (B(1), B(2), B(6), B(12), folate) in blood and plasma ghrelin level in these subjects were analyzed. Results showed that plasma level of ghrelin and lipid oxidation in NSCLC patients were significantly greater than control groups (P0.05). The activity of GPX, SOD, or catalase was significantly reduced, but XO activity was significantly elevated in NSCLC patients (P0.05). Plasma level of GSH was significantly lower, but GSSG level was significantly increased in NSCLC patients (P0.05). Vitamins B(2) and B(6) levels in red blood cells (RBC) from NSCLC patients were significantly lower (P0.05), and both were negatively correlated with plasma ghrelin. The correlation coefficients were -0.788 and -0.752, respectively. These data suggest that plasma GSH level may be a proper biomarker for evaluating oxidation status for NSCLC patients. RBC levels of vitamins B2 and B6 were reduced in NSCLC patients; thus, the importance of vitamins B(2) and B(6) for NSCLC patients could not be ignored.

Alleviative effects of histidine and carnosine in mice against ethanol-induced oxidative and inflammatory was examined. After chronic alcoholic liver injury was induced, histidine and carnosine at 0.5, 1, 2g/L were added to the drinking water for 3 weeks. Results showed that the post-intake of histidine or carnosine markedly decreased alanine aminotransferase and aspartate aminotransferase activities (P

According to the multiple alignment of various dihydrolipoamide dehydrogenases (E3s) sequences, three human mutant E3s of the conserved residues in the center domain, N286D, N286Q, and D320N were created, over-expressed and purified. We characterized these mutants to investigate the reaction mechanism of human dihydrolipoamide dehydrogenases. The specific activities of N286D, N286Q, and D320N are 30.84%, 24.57% and 48.60% to that of the wild-type E3 respectively. The FAD content analysis indicated that these mutant E3s about 96.0%, 99.4% and 82.7% of FAD content compared to that of wild-type E3 respectively. The molecular weight analysis showed that these three mutant proteins form the dimer. Kinetic's data demonstrated that the K(cat) of both forward and reverse reactions of these mutant proteins were decreased. These results suggest that N286 and D320 play a role in the catalytic function of the E3.