Calcitriol Research Articles

Parathyroid hormone serum concentration kinetic profile in critically ill patients undergoing continuous renal replacement therapies: a prospective observational study.

Elevated serum parathormone (PTH) levels have been observed in acute kidney injury and are related to calcium-phosphate metabolism disturbance, decreased renal production of 1,25 dihydroxyvitamin D3, impaired renal PTH excretion, and other renal-independent factors. There are no data regarding PTH concentration kinetics in critically ill patients undergoing continuous renal replacement therapies (CRRT) in an intensive care setting. The primary objective of this study was to investigate trends in PTH serum levels in critically ill patients with multiorgan failure undergoing CRRT, by performing periodic PTH measurements in the acute phase of critical illness. This was a single-centre, prospective, observational study conducted in an mixed, university-affiliated intensive care unit. Critically ill patients who fulfilled all of the following criteria were included: respiratory failure; circulatory failure; acute kidney injury treated by CRRT; and sequential organ failure assessment score (SOFA score) of 5 or more. Patients who met any of the following criteria were excluded: acute liver failure; hypercalcemia at admission (total calcium serum level > 10.6 mg/dL; total ionized calcium plasma level > 1.35 mmol/L); parathyroid gland disease; end-stage renal disease; patients undergoing therapeutic plasma exchange or extracorporeal membrane oxygenation procedures; aged under 18 years; pregnant; and life expectancy after admission to the intensive care unit anticipated to be less than 72 hours as assessed by the investigator. Thirty patients met the inclusion criteria. A statistically significant change in PTH over time was observed (Friedman ANOVA; p = 0.0001). The post-hoc test showed a statistically significant decrease in PTH: measurements 5-8 relative to measurement 1, and measurements 4-8 relative to measurement 2 (p < 0.05). No significant correlations between 25 hydroxyvitamin D3 deficiency, age, diagnosis, SOFA score, and PTH levels were observed. A statistical test indicated that serum concentrations of PTH were significantly higher in the de novo sepsis group (p < 0.05). The PTH serum concentration decreases during the course of CRRT in the majority of patients. When the course of the disease starts to be complicated by sepsis, PTH serum levels then remain high. A probable reason for this is the existence of the inflammatory state triggered by sepsis.

Vitamin D Enhances Radiosensitivity of Colorectal Cancer by Reversing Epithelial-Mesenchymal Transition.

Colorectal cancer (CRC) is often resistant to conventional therapies. Previous studies have reported the anticancer effects of vitamin D in several cancers, its role in radiotherapy (RT) remains unknown. We found that 1α, 25-dihydroxyvitamin D3 (VD3), the biologically active form of vitamin D, had antitumor effect on CRC and sensitized CRC cells to ionizing radiation (IR). VD3 demonstrated synergistic effect in combination with IR, which were detected by colony formation and cell proliferation assay. Radiosensitivity restoration induced by VD3 was associated with a series of phenotypes, including apoptosis, autophagy, and epithelial-mesenchymal transition (EMT). Using proteomics, “regulation of cell migration” and “cadherin” were found to be obviously enriched GO terms. Moreover, cystatin D and plasminogen activator inhibitor-1 (PAI-1), the differentially expressed proteins, were associated with EMT. Next, we confirmed the contributions of these two genes in enhancing IR sensitivity of CRC cells upon inhibition of EMT. As determined by proteomics, the mechanism underlying such sensitivity involved partially block of JAK/STAT3 signaling pathway. Furthermore, VD3 also elicited sensitization to RT in xenograft CRC models without additional toxicity. Our study revealed that VD3 was able to act in synergy with IR both in vitro and in vivo and could also confer radiosensitivity by regulating EMT, thereby providing a novel insight for elevating the efficacy of therapeutic regimens.

Clostridium butyricum alone or combined with 1, 25-dihydroxyvitamin D3 improved early-stage broiler health by modulating intestinal flora.

This study was conducted to investigate the effects of Clostridium butyricum in isolation or in combination with 1, 25-dihydroxyvitamin D3 in early-stage broilers. A total of 360 half male and half female Cobb broilers (1day old) were randomly divided into four groups: Con (basal diet), Anti (basal diet+75mg/kg chlortetracycline), Cb (basal diet+109 CFU per kg C. butyricum) and CD (basal diet+109 CFU per kg C. butyricum+25μg/kg 1,25(OH)2 D3 ). The results were as follows: (1) Compared with Con, CD significantly increased ADG (p<0.05). (2) Contrast with Con and Anti, Cb and CD significantly increased glutathione peroxidase and SOD in the serum and liver, and decreased malondialdehyde content in serum (p<0.05). (3) In addition, the content of immunoglobulin (IgA, IgY and IgM) in Cb and CD birds was higher than that in Con birds (p<0.05); the Cb supplementation decreased (p<0.05) the contents of IL-8, IL-1β and TNF-α than those in Con. (4) Cb and CD had lower caecal acetic and propionic content than the Anti group (p<0.05). (5) The community richness of Con was significantly higher than that of Anti (p<0.05). The relative abundance of Alistipes and Ruminococcaceae-UCG-014 in Cb and CD supplemented birds were lower than those in Con (p<0.05). The relative abundant of Escherichia-Shigella in CD was higher than Con and Anti (p<0.05). These data indicated that dietary C. butyricum and 1, 25-dihydroxyvitamin D3 can improve the growth performance, immunity responses, antioxidation, bone development and intestinal microflora in early-stage broilers. Oral administration of C. butyricum or C. butyricum combined with 1,25-dihydroxyvitamin D3 enhanced immunity and antioxidant activity in early-stage birds.

Relationship between vitamin D receptor gene FokI polymorphism and 25-hydroxyvitamin D levels in apparently healthy Syrians

The vitamin D receptor (VDR) protein has an essential function in regulating serum calcium and 1, 25 dihydroxy vitamin D3. Therefore, variations in VDR gene play an essential role in vitamin D levels.This study aimed to detect the distribution of FokI polymorphism in healthy Syrians and its relation to 25-OH-vitamin D levels.The study included 197 unrelated healthy individuals (Female:131 and Male:66) divided into four groups according to their vitamin D levels: no hypovitaminosis D(G1), vitamin D insufficiency(G2), vitamin D deficiency (G3), and severe vitamin D deficiency (G4). Genotyping for the FokI single nucleotide polymorphism was performed by PCR followed by restriction fragment length polymorphism analysis (RFLP).Frequencies of FF, Ff and ff genotypes were 32.4%, 50% and 17.6%, in G1, 19.6%, 60.8% and 19.6% in G2, 52.1%, 37.5% and 10.4% in G3, and 58%, 37.7% and 4.3% in G4 respectively, with significant differences between the four groups.In conclusion, this study showed a strong correlation between 25-OH-vitamin D levels and FokI polymorphism of the VDR gene in apparently healthy Syrians.

High-dose vitamin D3 supplementation ameliorates renal fibrosis by vitamin D receptor activation and inhibiting TGF-β1/Smad3 signaling pathway in 5/6 nephrectomized rats

Renal fibrosis is the pathological consequence of progressive chronic kidney disease. Although it has been reported that vitamin D3 exerts antifibrotic effects, the underlying mechanisms remain unclear. This study is aimed at investigating the effects and molecular mechanisms in high-dose vitamin D3 treatment on renal fibrosis. A model of chronic kidney disease was established by 5/6 nephrectomy in rats characterised by high levels of serum creatine, urea nitrogen, and urinary protein. Serum 25-dihydroxyvitamin D3, calcium and parathormone levels were measured to evaluate vitamin D levels. Hematoxylin and eosin, periodic acid Schiff and Mallory's Trichrome staining were used to evaluate histopathological changes in rats. Moreover, the expression of vimentin, collagen I, α-smooth muscle actin and E-cadherin were analyzed at molecular and histopathological levels. Our results showed that exposure to vitamin D3 decreased the levels of serum creatine, urea nitrogen and urine protein and restored the homeostasis of calcium and parathormone. Vitamin D3 also downregulated the expression of vimentin, collagen I and α-smooth muscle actin and attenuated renal fibrosis and epithelial to mesenchymal transition in the kidney. Importantly, vitamin D3 treatment increased the expression of the vitamin D receptor and inhibited Transforming growth factor-β1 (TGF-β1)/Smad3 signaling pathway in rats kidneys with chronic kidney disease. Mechanistically, the upregulation of TGF-β1 and phosphorylation of Smad3 induced by vitamin D3 was reversed by activation of the vitamin D receptor. Our findings indicated that vitamin D3 is a potential antifibrotic drug in chronic kidney disease via the vitmin D receptor and inhibiting TGF-β1/Smad3 signaling pathway.

Impaired 1,25 dihydroxyvitamin D3 action and hypophosphatemia underlie the altered lacuno-canalicular remodeling observed in the Hyp mouse model of XLH.

Osteocytes remodel the perilacunar matrix and canaliculi. X-linked hypophosphatemia (XLH) is characterized by elevated serum levels of fibroblast growth factor 23 (FGF23), leading to decreased 1,25 dihydroxyvitamin D3 (1,25D) production and hypophosphatemia. Bones from mice with XLH (Hyp) have enlarged osteocyte lacunae, enhanced osteocyte expression of genes of bone remodeling, and impaired canalicular structure. The altered lacuno-canalicular (LCN) phenotype is improved with 1,25D or anti-FGF23 antibody treatment, pointing to roles for 1,25D and/or phosphate in regulating this process. To address whether impaired 1,25D action results in LCN alterations, the LCN phenotype was characterized in mice lacking the vitamin D receptor (VDR) in osteocytes (VDRf/f;DMP1Cre+). Mice lacking the sodium phosphate transporter NPT2a (NPT2aKO) have hypophosphatemia and high serum 1,25D levels, therefore the LCN phenotype was characterized in these mice to determine if increased 1,25D compensates for hypophosphatemia in regulating LCN remodeling. Unlike Hyp mice, neither VDRf/f;DMP1Cre+ nor NPT2aKO mice have dramatic alterations in cortical microarchitecture, allowing for dissecting 1,25D and phosphate specific effects on LCN remodeling in tibial cortices. Histomorphometric analyses demonstrate that, like Hyp mice, tibiae and calvariae in VDRf/f;DMP1Cre+ and NPT2aKO mice have enlarged osteocyte lacunae (tibiae: 0.15±0.02μm2(VDRf/f;DMP1Cre-) vs 0.19±0.02μm2(VDRf/f;DMP1Cre+), 0.12±0.02μm2(WT) vs 0.18±0.0μm2(NPT2aKO), calvariae: 0.09±0.02μm2(VDRf/f;DMP1Cre-) vs 0.11±0.02μm2(VDRf/f;DMP1Cre+), 0.08±0.02μm2(WT) vs 0.13±0.02μm2(NPT2aKO), p<0.05 all comparisons) and increased immunoreactivity of bone resorption marker Cathepsin K (Ctsk). The osteocyte enriched RNA isolated from tibiae in VDRf/f;DMP1Cre+ and NPT2aKO mice have enhanced expression of matrix resorption genes that are classically expressed by osteoclasts (Ctsk, Acp5, Atp6v0d2, Nhedc2). Treatment of Ocy454 osteocytes with 1,25D or phosphate inhibits the expression of these genes. Like Hyp mice, VDRf/f;DMP1Cre+ and NPT2aKO mice have impaired canalicular organization in tibia and calvaria. These studies demonstrate that hypophosphatemia and osteocyte-specific 1,25D actions regulate LCN remodeling. Impaired 1,25D action and low phosphate levels contribute to the abnormal LCN phenotype observed in XLH.

Effects of 1,25 Dihydroxyvitamin D3 on Human Retinal Pigment Epithelial Cell Lines

To assess the effects of 1,25 dihydroxyvitamin D3 (vitamin D3) either alone or under oxidative damage on human retinal pigment epithelium cell lines. The human retinal pigment epithelial cell lines were pretreated with hydrogen peroxide with different concentrations (100-1000μM) and durations (4, 12 and 24h) to determine the appropriate dose. A group of cells were treated with vitamin D3 alone, and another group of cells were co-treated with different concentrations of (10-100nM) vitamin D3 and hydrogen peroxide. Anti-cytotoxic, anti-apoptotic and anti-genotoxic effects of vitamin D3 on the hydrogen peroxide treated cell line were evaluated. In addition, mitochondrial membrane potentials of treated cell lines were measured. Vitamin D3 showed statistically significant anti-cytotoxic effects and increased cell viability in all concentrations (p < 0.001). It has also significantly decreased the intracellular ROS generation at concentrations between 10-60nM and increased intracellular reactive oxygen species in high doses over 90nM (p < 0.01). When apoptosis was evaluated, vitamin D3 caused statistically significant decrease in a dose-dependent manner (p < 0.001). In terms of DNA damage which was caused by oxidative stress, it was observed that vitamin D3 significantly reduced the damage in a dose-dependent manner (p < 0.001). At the doses of 10-50nM, vitamin D3 significantly decreased the mitochondrial membrane potential (p < 0.01). Our study suggests that 1,25 (OH)2 D3 is capable for alleviating the oxidative damage in ARPE cell lines. With these results, vitamin D is thought to be a therapeutic alternative for the prevention of age-related macular degeneration. This warrants further investigations.

Protective Effects of 1,25 Dihydroxyvitamin D3 against High-Glucose-Induced Damage in Human Umbilical Vein Endothelial Cells Involve Activation of Nrf2 Antioxidant Signaling

Aim: To explore the protective effects and related mech­anisms of 1,25 dihydroxyvitamin D₃ (1,25(OH)₂D₃) on en­dothelial dysfunction under hyperglycemic conditions. Methods: Cultured human umbilical vein endothelial cells (HUVECs) were treated with normal glucose (glucose concentration of 5.5 mmol/L), high glucose (glucose concentration of 33 mmol/L), and high glucose plus 1,25(OH)₂D₃, respectively. Cell viability and apoptosis, intracellular reactive oxygen species (ROS) and nitric oxide (NO) contents, antioxidant enzyme activities, proinflammatory cytokine mRNA levels, and expression levels of proteins involved were measured. Results: High glucose decreased HUVEC viability, promoted ROS production and apoptosis, and reduced NO generation, which was associated with decreased activities of antioxidant enzymes and increased levels of proinflam­matory cytokines. 1,25(OH)₂D₃ treatment enhanced HUVEC viability, attenuated ROS generation and apoptosis, and ­increased NO production, which was accompanied by ­enhanced antioxidant enzyme activities and reduced ­proinflammatory factors. Mechanically, 1,25(OH)₂D₃ promoted nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) in a vitamin D receptor (VDR)-dependent manner, and Nrf2 siRNA abolished the antioxidative and ­anti-inflammatory effects of 1,25(OH)₂D₃. Conclusions: 1,25(OH)₂D₃ attenuates high-glucose-induced endothelial oxidative injury through upregulation of the Nrf2 antioxidant pathway in a VDR-dependent manner.

Cyanocobalamin and/or calcitriol mitigate renal damage-mediated by tamoxifen in rats: Implication of caspase-3/NF-κB signaling pathways

AimTamoxifen (TAMO) is a chemotherapeutic drug used for the treatment of breast cancer. Nevertheless, there is a lack of information available in regarding its nephrotoxicity. The purpose of this work was to investigate the impact of cyanocobalamin (COB) and/or calcitriol (CAL) injections on TAMO-induced nephrotoxicity. Main methodsAnimals were allocated into five groups as follows: normal control group; TAMO (45 mg/kg) administered group; TAMO+COB (6mg/kg, i.p) treated group; TAMO+CAL (0.3 μg/kg, i.p) treated group; TAMO+COB+CAL combination groups. Key findingsRenal injury induced by TAMO was confirmed by the alteration in renal function parameters in the serum (urea and creatinine), as well as in the urine (creatinine clearance, total protein and albumin). These results were supported by histopathological examination. Upregulation of renal inflammatory parameters; tumor necrosis factor (TNF)-α, interleukin (IL)-6, C-reactive protein (CRP); and transforming growth factor (TGF)-β1 as well as in protein expression of nuclear factor-kappa B (NF-κB) and cleaved caspase-3 were observed to a greater extent in the TAMO-treated rats compared with the control. Renal fibrosis was also evidenced by a elevation in renal L-hydroxyproline level as well as by histomorphological collagen deposition in TAMO-treated groups compared to the control group. Administration of COB and/or CAL concurrently with TAMO significantly ameliorated the deviation in the above-studied parameters and improved the histopathological renal picture. SignificanceInhibition of NF-κβ-mediated inflammation and caspase-3-induced apoptosis are possible renoprotective mechanisms of COB and/or CAL against TAMO nephrotoxicity, which was more noticeable in the TAMO group treated with the combination of the two vitamins in question.

Reproductive performance, expression of TRAP6 and TGF-β4 genes in utero-vaginal junction mucosa, and indicators of liver function in female Chukar partridge (Alectoris chukar) breeders fed with fish oil and calcitriol during the egg-laying period

Reproductive attributes, expression of TRAP6 and TGF-β mRNA in the mucosa of the utero-vaginal junction (UVJ) of oviduct, and liver function were evaluated in Chukar partridge (Alectoris chukar) breeders subjected to long-term oral administration of fish oil (FO) and/or calcitriol (CT). A total of forty-eight 1.5-year-old laying Chukar partridges and 16 age-matched males (female:male ratio of 3:1) were randomly allocated to four groups (4 replicates of 3 female birds and one male bird each). Breeder females in groups 1, 2, and 3 were orally administered daily with 0.2 mL (0.24 g)/500 g body weight FO, 0.2 mL solution containing 10 μg CT, or their combination (FO + CT) for 42 successive days, respectively. Pure crystalline calcitriol was dissolved in ethanol (30%) prior to administration. The control group (CON), received a similar volume of a 30% solution of ethanol only. Eggs were collected and incubated to evaluate the reproductive performance. Blood samples were taken on days 0, 21, and 42 of the trial for the quantification of serum alkaline phosphatase (ALP), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). On day 43, one bird per replicate was killed by cervical dislocation to assess the expression of TRAP6 and TGF-β genes in the UVJ mucosa. Administration of CT or FO + CT increased the egg production rate, fertility rate, and hatchability rate of the set eggs. Fertility duration and sperm penetration rate were higher in partridges receiving FO and (or) CT, but chick quality, and embryonic mortality were not affected by the treatment effect. Administration of CT or FO + CT decreased the serum ALT and AST levels. Administration of FO or CT was associated with a lower expression of TGF-β mRNA in the UVJ mucosa. Oral administration of FO resulted in a reduction in the expression of TRAP6 in the UVJ mucosa. However, the birds fed with CT or FO + CT recorded a higher mRNA expression for TRAP6. Although the reproductive performance and TRAP6 expression were higher following the feeding of FO or FO + CT, expression of TGF-β was decreased, suggesting plausibly that TGF-β may not have a determinant effect on the reproductive attributes in female Chukar partridges. Further studies are required to understand the mechanisms underlying the effects of TRAP6 and TGF-β on other reproductive criteria in partridges.

Adventitial delivery of nanoparticles encapsulated with 1\u03b1, 25-dihydroxyvitamin D3 attenuates restenosis in a murine angioplasty model

Percutaneous transluminal angioplasty (PTA) of stenotic arteriovenous fistulas (AVFs) is performed to maintain optimal function and patency. The one-year patency rate is 60% because of venous neointimal hyperplasia (VNH) and venous stenosis (VS) formation. Immediate early response gene X-1 (Iex-1) also known as Ier3 increases in response to wall shear stress (WSS), and can cause VNH/VS formation in murine AVF. In human stenotic samples from AVFs, we demonstrated increased gene expression of Ier3. We hypothesized that 1α, 25-dihydroxyvitamin D3, an inhibitor of IER3 delivered as 1α, 25-dihydroxyvitamin D3 encapsulated in poly lactic-co-glycolic acid (PLGA) nanoparticles loaded in Pluronic F127 hydrogel (1,25 NP) to the adventitia of the stenotic outflow vein after PTA would decrease VNH/VS formation by reducing Ier3 and chemokine (C–C motif) ligand 2 (Ccl2) expression. In our murine model of AVF stenosis treated with PTA, increased expression of Ier3 and Ccl2 was observed. Using this model, PTA was performed and 10-μL of 1,25 NP or control vehicle (PLGA in hydrogel) was administered by adventitial delivery. Animals were sacrificed at day 3 for unbiased whole genome transcriptomic analysis and at day 21 for immunohistochemical analysis. Doppler US was performed weekly after AVF creation. At day 3, significantly lower gene expression of Ier3 and Ccl2 was noted in 1,25 NP treated vessels. Twenty-one days after PTA, 1,25 NP treated vessels had increased lumen vessel area, with decreased neointima area/media area ratio and cell density compared to vehicle controls. There was a significant increase in apoptosis, with a reduction in CD68, F4/80, CD45, pro-inflammatory macrophages, fibroblasts, Picrosirius red, Masson’s trichrome, collagen IV, and proliferation accompanied with higher wall shear stress (WSS) and average peak velocity. IER3 staining was localized to CD68 and FSP-1 (+) cells. After 1,25 NP delivery, there was a decrease in the proliferation of α-SMA (+) and CD68 (+) cells with increase in the apoptosis of FSP-1 (+) and CD68 (+) cells compared to vehicle controls. RNA sequencing revealed a decrease in inflammatory and apoptosis pathways following 1,25 NP delivery. These data suggest that adventitial delivery of 1,25 NP reduces VNH and venous stenosis formation after PTA.

Vitamin D receptor expression in mature osteoclasts reduces bone loss due to low dietary calcium intake in male mice

Mature osteoclasts express the vitamin D receptor (VDR) and are able to respond to active vitamin D (1α, 25-dihydroxyvitamin D3; 1,25(OH)2D3) by regulating cell maturation and activity. However, the in vivo consequences of vitamin D signalling directly within functionally mature osteoclasts is only partially understood. To investigate the in vivo role of VDR in mature osteoclasts, conditional deletion of the VDR under control of the cathepsin K promoter (CtskCre/Vdr−/−), was assessed in 6 and 12-week-old mice, either under normal dietary conditions (NormCaP) or when fed a low calcium (0.03 %), low phosphorous (0.08 %) diet (LowCaP). Splenocytes from CtskCre/Vdr−/− mice were co-cultured with MLO-Y4 osteocyte-like cells to assess the effect on osteoclastogenesis. Six-week-old CtskCre/Vdr-/- mice demonstrated a 10 % decrease in vertebral bone volume (p < 0.05), which was associated with increased osteoclast size (p < 0.05) when compared to Vdrfl/fl control mice. Control mice fed a LowCaP diet exhibited extensive trabecular bone loss associated with increased osteoclast surface, number and size (p < 0.0001). Interestingly, CtskCre/Vdr-/- mice fed a LowCaP diet showed exacerbated loss of bone volume fraction (BV/TV%) and trabecular number (Tb.N), by a further 22 % and 21 %, respectively (p < 0.05), suggesting increased osteoclastic bone resorption activity with the loss of VDR in mature osteoclasts under these conditions. Co-culture of CtskCre/Vdr-/- splenocytes with MLO-Y4 cells increased resulting osteoclast numbers 2.5-fold, which were greater in nuclei density and exhibited increased resorption of dentine compared to osteoclasts derived from Vdrfl/fl splenocyte cultures. These data suggest that in addition to RANKL-mediated osteoclastogenesis, intact VDR signalling is required for the direct regulation of the differentiation and activity of osteoclasts in both in vivo and ex vivo settings.

Cadmium Toxicity on Chondrocytes and the Palliative Effects of 1α, 25-Dihydroxy Vitamin D3 in White Leghorns Chicken's Embryo.

Cadmium (Cd) can causes osteoporosis and joint swelling. However, the mechanism of Cd toxicity in chondrocytes and how to alleviate Cd poisoning to chondrocytes are still unclear. Herein, we evaluated the toxicity of Cd to chicken chondrocytes, and whether vitamin D can relieve the toxicity of Cd to chondrocytes. Primary chondrocytes were collected from knee-joint cartilage of 15-day-old chicken embryos. They were treated with (0, 1, 2, and 4) μM Cd alone, 10−8 M 1α,25-(OH)2D3 alone, or 2 μM Cd combined with 10−8 M 1α,25-(OH)2D3. We found that Cd significantly inhibited Sox9 and ACAN mRNA expression, which are markers for chondrocyte differentiation, downregulated the mitochondrial membrane potential, upregulated the Bax/B-cell lymphoma 2 ratio. Furthermore, Cd significantly promoted matrix metalloproteinase (MMP)-9 expression, thus accelerating the degradation of extracellular matrix. And Cd also inhibited the expression of main macromolecular protein of extracellular matrix, Collagen type IIα1 (COL2A1) and acid mucopolysaccharide. However, 1α,25-(OH)2D3 pretreatment significantly alleviated the toxicity effects of Cd on the differentiation, apoptosis and extracellular matrix gene expression in primary chondrocytes. Conclusively, Cd exposure could inhibited chicken embryo chondrocytes differentiation, extracellular matrix gene expression, and induced chondrocyte apoptosis. However, these toxic effects of Cd are alleviated by the pretreatment of chondrocytes with 1α,25-(OH)2D3.

A Double-Blind Randomized Controlled Trial Comparing the Efficacy of 0.0003% Calcitriol with 1% Pimecrolimus versus Placebo in the Management of Pityriasis Alba

A Double-Blind Randomized Controlled Trial Comparing the Efficacy of 0.0003% Calcitriol with 1% Pimecrolimus versus Placebo in the Management of Pityriasis Alba

Role of 1, 25-dihydroxyvitamin D3 in liver lipid metabolism induced by methionine-choline-deficient diet in rats

Objective: To investigate the role of 1, 25-dihydroxyvitamin D3 [1.25(OH) (2)D(3)] in liver lipid metabolism so as to provide the clues for elucidating the mechanism of non-alcoholic fatty liver. Methods: 26 SD rats were randomly divided into control group (methionine-choline-sufficient diet, MCS), model group (methionine-choline-deficiency diet, MCD) and intervention group [MCD+1.25(OH) (2)D(3)]. The intervention, control, and model group was given 3 ng/100 g 1.25(OH) (2)D(3) peanut oil solution per day by gavage according to body mass. After 4 weeks the experiment was ended up, and the blood was collected from the inferior vena cava to detect alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The liver tissue was collected to observe the liver morphological and pathological changes (oil red O and HE staining). The changes in the level of liver total triglyceride (TG) content and liver lipid metabolism-related genes [fatty acid transfer protein (FAT/CD36), acetyl-coenzyme A carboxylase (ACC1)] mRNA and protein were detected. One-way analysis of variance was used to compare the means between groups. Results: Oil red O staining and HE staining showed that lipid droplet-vacuoles were significantly increased in the liver tissue of the model group than that of the intervention group. The liver TG content (2.23 ± 0.98) μmol/g of the intervention group was significantly lower than that of the model group (3.53 ± 1.06) μmol/g (F = 5.930, P = 0.035). The ALT content of the intervention group (35.99±9.54) U/L was significantly lower than that of the model group (57.65 ± 19.42) U/L (F = 13.790, P = 0.034). The AST content of the intervention group (16.9 ± 3.73) U/L was significantly lower than that of the model group (27.81 ± 13.31) U/L (F = 3.084, P = 0.046). The relative expression levels of mRNA and protein (mRNA: 1.21 ± 0.61, protein: 1.54 ± 0.75) of FAT/CD36 in the intervention group were significantly lower than those of the model group (mRNA: 2.31 ± 0.81, protein: 2.83 ± 1.42) (mRNA: F = 8.370, P = 0.001, protein: F = 7.212, P = 0.043). The relative expression level of mRNA and protein of ACC1 (mRNA: 0.89 ± 0.54, protein: 0.28 ± 0.11) were also significantly lower than those in model group (mRNA: 1.39 ± 0.19, protein: 0.47 ± 0.24) (mRNA: F = 3.948, P = 0.036, protein: F = 10.933, P = 0.048). Conclusion: 1.25(OH) (2)D(3) can reduce liver fat deposition in rats fed with MCD by inhibiting the expression of fat / CD36 and ACC1.

Fucoidan Ameliorates Renal Injury-Related Calcium-Phosphorus Metabolic Disorder and Bone Abnormality in the CKD-MBD Model Rats by Targeting FGF23-Klotho Signaling Axis.

Background: Recently, chronic kidney disease (CKD)-mineral and bone disorder (MBD) has become one of common complications occurring in CKD patients. Therefore, development of a new treatment for CKD–MBD is very important in the clinic. In China, Fucoidan (FPS), a natural compound of Laminaria japonica has been frequently used to improve renal dysfunction in CKD. However, it remains elusive whether FPS can ameliorate CKD–MBD. FGF23-Klotho signaling axis is reported to be useful for regulating mineral and bone metabolic disorder in CKD–MBD. This study thereby aimed to clarify therapeutic effects of FPS in the CKD–MBD model rats and its underlying mechanisms in vivo and in vitro, compared to Calcitriol (CTR). Methods: All male rats were divided into four groups: Sham, CKD–MBD, FPS and CTR. The CKD–MBD rat models were induced by adenine administration and uninephrectomy, and received either FPS or CTR or vehicle after induction of renal injury for 21 days. The changes in parameters related to renal dysfunction and renal tubulointerstitial damage, calcium-phosphorus metabolic disorder and bone lesion were analyzed, respectively. Furthermore, at sacrifice, the kidneys and bone were isolated for histomorphometry, immunohistochemistry and Western blot. In vitro, the murine NRK-52E cells were used to investigate regulative actions of FPS or CTR on FGF23-Klotho signaling axis, ERK1/2-SGK1-NHERF-1-NaPi-2a pathway and Klotho deficiency. Results: Using the modified CKD–MBD rat model and the cultured NRK-52E cells, we indicated that FPS and CTR alleviated renal dysfunction and renal tubulointerstitial damage, improved calcium-phosphorus metabolic disorder and bone lesion, and regulated FGF23-Klotho signaling axis and ERK1/2-SGK1-NHERF-1-NaPi-2a pathway in the kidney. In addition, using the shRNA-Klotho plasmid-transfected cells, we also detected, FPS accurately activated ERK1/2-SGK1-NHERF-1-NaPi-2a pathway through Klotho loss reversal. Conclusion: In this study, we emphatically demonstrated that FPS, a natural anti-renal dysfunction drug, similar to CTR, improves renal injury-related calcium-phosphorus metabolic disorder and bone abnormality in the CKD–MBD model rats. More importantly, we firstly found that beneficial effects in vivo and in vitro of FPS on phosphorus reabsorption are closely associated with regulation of FGF23-Klotho signaling axis and ERK1/2-SGK1-NHERF-1-NaPi-2a pathway in the kidney. This study provided pharmacological evidences that FPS directly contributes to the treatment of CKD–MBD.

Smell Regions in Patients with Vitamin D Deficiency: An MRI Evaluation.

Objective We investigated the effects of vitamin D deficiency in the peripheral and central smell regions by magnetic resonance imaging (MRI). Methods This retrospective study included 29 patients (12 males, 17 females) with 25-dihydroxy vitamin D3 [25(OH) 2 D 3 ] deficiency (group 1) and 34 subjects without 25(OH) 2 D 3 deficiency (14 males, 20 females) (group 2). Using cranial MRIs, the peripheral (olfactory bulb [OB] volume and olfactory sulcus [OS] depth) and central (insular gyrus and corpus amygdala) smell regions were evaluated. Results The OB volume and OS depth values of the 25(OH) 2 D3 deficiency group were significantly lower than those of the control group ( p < 0.05). For the central smell regions, the insular gyrus and corpus amygdala areas of the 25(OH) 2 D3 deficiency group were nonsignificantly lower than those in the control group ( p > 0.05). There were positive correlations between OB volumes, OS depths, and insular gyrus and corpus amygdala areas bilaterally in the 25(OH) 2 D3 deficiency group separately and in all subjects (groups 1 and 2) ( p < 0.05). In the 25(OH) 2 D3 deficiency group, as the 25(OH) 2 D3 values became lower, the insular gyrus area values decreased bilaterally ( p < 0.05). In females, the corpus amygdala area values were lower than in males ( p < 0.05). Conclusion Since vitamin D3 deficiency affected the peripheral and central smell regions negatively, we recommend evaluating patients' vitamin D levels as a health policy to prevent vitamin D3 deficiency-related cranial smell region problems. Moreover, sunlight exposure is very important to increase vitamin D levels, and the public should be informed about this topic.

Influence of the Biofield Energy Treatment on the Pharmacokinetics of 25-Hydroxyvitamin D3 in Male Sprague-Dawley Rats after a Single Oral Dose of Vitamin D3

Vitamin D3 reported having many important roles in bone metabolism, osteoporosis, immunity, and useful in numerous diseases, i.e., cardiovascular, cancers, and neurodegenerative diseases. Unexpectedly, there is little information available about the concentration of 25-hydroxyvitamin D3 in the blood, which is the best indicator of vitamin D3 status after its oral absorption. However, the biological activity of vitamin D3 is mediated via the formation of the active metabolite, 1-α, 25-dihydroxyvitamin D3 . There are several factors that affect its absorption and first-pass metabolism that lead to having very low plasma concentrations of both the metabolites (25-hydroxyvitamin D3 and 1-α, 25-dihydroxyvitamin D3 ) following an oral administration. Thus, the current study was executed to determine the influence of the Trivedi Effect®-Consciousness Energy Treatment on vitamin D3 and rats through the measurement of plasma 25-hydroxyvitamin D3 concentrations after the oral administration of vitamin D3 in male Sprague-Dawley rats. The test item, vitamin D3 was divided into two parts. One part was denoted as the control (without Biofield Energy Treatment/Blessing), while the other part was defined as the Biofield Energy Treated sample, which received the Biofield Energy Treatment by renowned Biofield Energy Healer, Dahryn Trivedi. Additionally, one group of animals also received Consciousness Energy Treatment per se by the Healer under similar conditions. The Biofield Energy Healer who was located in the USA, while the test samples and animals were located in the research laboratory in India. Vitamin D3 oral formulations were administrated by oral gavage at a dose of 500 µg per kg in groups viz. G1 (untreated vitamin D3 ), G2 (Biofield Treated vitamin D3 ), and G3 (Biofield Treated animals received untreated vitamin D3 ) group. The Biofield Energy Treatment increased the maximum plasma concentration (Cmax) of 25-hydroxyvitamin D3 by 3.45% and 72.77% in the G2 and G3 groups, respectively as compared with the G1 group. The area under the plasma concentration– time curve (AUC0–t) of 25-hydroxyvitamin D3 was altered by -11.19% in the G2 group and 59.45% in the G3 group as compared to the G1 group. After oral administration, the Tmax of 25-hydroxyvitamin D3 was altered by -62.97% in G2 and 55.56% in G3 groups compared to G1. The mean residence time (MRT) of 25-hydroxyvitamin D3 was also altered in the G2 group by -12.34% and G3 group by 0.18%, as compared to the G1. The relative oral bioavailability (Fr) of 25-hydroxyvitamin D3 was significantly altered by -11.19% in the G2 group and 59.45% in the G3 group compared to the G1. Biofield Energy Treatment could be an innovative strategy that opens new avenues to overcome poorly absorbed pharmaceuticals/ nutraceuticals/herbal extracts and can also improve the therapeutic performance of orally active molecules.

Effect of Vitamin D Adjuvant and Allergen Specific Immunotherapy on Serum IL-10 and IL-17 Levels in Childhood Asthma: A Controlled Clinical Trial

Background: 1, 25-dihydroxy vitamin D3 (VitD3) can improve the effect of allergenspecific immunotherapy (SIT). Few data is available about its role in childhood asthma. Objective: To assess the immunological and clinical efficacy of VitD3 as an adjuvant to allergen specific immunotherapy in pediatric asthma. Methodology: Sixty nine children with atopic asthma were divided into three groups: a group received subcutaneous immunotherapy (SCIT) in combination with VitD3 (n=23), another group received SCIT alone (n=23), and the last group VitD3 alone (n=23). All children were assessed at baseline, and six months for rate of inhaled corticosteroid (ICS) discontinuation, and serum levels of IL-10, and IL-17A. Results: In the SCIT + vitD3, ICS discontinuation rate was higher compared to VitD3 alone group and SCIT alone group at the end of 6th month (P=0.555 and 0.016 respectively). The combined SCIT+ VitD3 group showed significant increase of serum IL-10 level in comparison to SCIT alone group and VitD3 alone group (P=0.000) and significant decrease in serum IL-17A level compared to VitD3 alone group (P= 0.011) Conclusion: VitD3 enhance the clinical and immunological outcomes of SIT in pediatric asthma. Further investigation is needed to evaluate this effect in a larger scale to confirm its role as an adjunct to SIT.

Disorders of Phosphate Metabolism: Hypophosphatemia and Hyperphosphatemia

Phosphorus in the body exists as phosphate. Phosphate is the most abundant intracellular anion and exists mostly as organic phosphate compounds critical to cellular functions such as adenosine triphosphate (ATP). The kidneys are the main regulator of phosphate homeostasis. There are three hormonal systems responsible for phosphate homeostasis, the parathyroid hormone (PTH), Fibroblast growth factor-23 (FGF-23)/klotho, and 1,25 dihydroxyvitamin D3 (1,25 (OH)2D3, calcitriol). Most cases of hypophosphatemia are acquired and are due to malnutrition as in alcoholism. The kidneys maintain phosphate level in the normal range; therefore, hyperphosphatemia is uncommon until glomerular filtration rate (GFR) falls below 30 ml/min. Hyperphosphatemia is common in patients on renal replacement therapy and is treated with dietary phosphate restriction, and phosphate binders.