OBJECTIVE: To investigate the effect of vitamin D3 on polarization of monocyte macrophages induced by serum from patients with ankylosing spondylitis (AS). METHODS: Twenty AS naïve patients and 20 healthy controls from Wenzhou People's Hospital during January 2016 and December 2017 were enrolled. The macrophages were differentiated from THP1 cells induced by phorbol 12-myristate 13-acetate (PMA), and then co-cultured with the serum from healthy subjects (control group) or AS patients. Vitamin D3 was added in the medium mixed with serum from AS patients. Flow cytometry was used to analyze the ratio of CD68 and CD206 positive cells, and RT-PCR was performed to detect the mRNA expression of inducible nitric oxide synthase(iNOS) and arginase-1(Arg-1). RESULTS: THP1 cells could be polarized into mononuclear-macrophages with the induction of PMA. The proportion of CD206 positive cells in AS-serum group was lower than that in the control group (=9.434, <0.05), while the proportion of CD68 positive cells was higher than that in the control group (=43.920, <0.05). The proportion of CD206 positive cells in vitamin D3 group was higher than that in AS-serum group (=8.895, <0.05), while the proportion of CD68 positive cells was lower than that in AS-serum group (=9.089, <0.05). mRNA expression of Arg-1 in AS-serum group was lower than that in the control group (=8.899, <0.05), while mRNA expression of iNOS was higher than that in the control group (=3.656, <0.05). mRNA expression of Arg-1 in vitamin D3 group was higher than that in AS-serum group (=6.219, <0.05), while mRNA expression of iNOS was lower than that in AS-serum group (=5.876, <0.05). CONCLUSIONS Vitamin D3 can regulate the polarization of mononuclear macrophages for immunoregulation in patients with AS.
Adjuvants, Immunologic ; pharmacology ; Cell Differentiation ; drug effects ; Cholecalciferol ; pharmacology ; Humans ; Monocytes ; drug effects ; Spondylitis, Ankylosing ; blood ; physiopathology

OBJECTIVE: To investigate the effect of vitamin D3 to platelet activation by tumor cell culture medium. METHODS: The peripheral blood platelets of BALB/c mice were isolated. The platelets were activated in 4T1 culture fluid for 24 h. The platelets were divided into 7 groups: control group, activation group, 1 nmol/L vitamin D3 group, 10 nmol/L vitamin D3 group, 50 nmol/L vitamin D3 group, 100 nmol/L vitamin D3 group, and positive drug (0.1 μmol/L eptifibatide) group. CCK-8 assay was used to detect the platelet proliferation at 24, 48 and 72 h. Flow cytometry was used to detect the expression of CD61 and CD62p and receptor for advanced glycation end products (RAGE) at 24, 48 and 72 h. ELISA was used to detect the level of platelet-endothelial cell adhesion molecule-1 (PECAM-1) at 24, 48 and 72 h. RESULTS: The CD41 CONCLUSION Vitamin D3 shows antiplatelet effect and can inhibit platelet proliferation and activation.
Animals ; Blood Platelets ; Cell Culture Techniques ; Cholecalciferol/pharmacology* ; Flow Cytometry ; Mice ; Mice, Inbred BALB C ; P-Selectin ; Platelet Activation

OBJECTIVES: To investigate the level of 25 hydroxyvitamin D [25(OH)D] in late preterm infants and the effect of vitamin D₃ supplementation on the neurobehavioral development of infants and young children. METHODS: In this prospective study, 161 late preterm infants who were admitted from June 2017 to June 2020 were enrolled. According to the level of 25(OH)D in umbilical cord blood, they were divided into three groups: sufficiency group (n=52), insufficiency group (n=53), and deficiency group (n=56). Each group was further divided into subgroup A (vitamin D₃ 800 IU/d) and subgroup B (individualized vitamin D₃ supplementation) using a random number table. The levels of 25(OH)D were measured at 3 months after birth and at the corrected ages of 10 months and 18 months. The neurobehavioral development levels were determined by the Gesell Developmental Scale at the corrected ages of 10 months and 18 months. RESULTS: Within 24 hours and 3 months after birth, the insufficiency group and the deficiency group had a significantly lower level of 25(OH)D than the sufficiency group (P<0.05), and the insufficiency group had a significantly higher level of 25(OH)D than the deficiency group (P<0.05). In the deficiency group, subgroup B had a significantly higher level of 25(OH)D than subgroup A (P<0.05) at 3 months after birth. At the corrected ages of 10 months and 18 months, the insufficiency and deficiency groups had significantly lower scores of five functional areas of the Gesell Development Scale than the sufficiency group (P<0.05). Compared with the insufficiency group, the deficiency group had a significantly lower score of language at the corrected age of 10 months and a significantly lower score of gross motor at the corrected age of 18 months (P<0.05). Compared with subgroup A of the deficiency group, subgroup B had a significantly higher score of adaptive ability at the corrected age of 10 months and significantly higher scores of adaptive ability and response ability at the corrected age of 18 months (P<0.05). CONCLUSIONS There is a significant difference in the level of 25(OH)D in umbilical cord blood in late preterm infants. Individualized vitamin D supplementation appears to be more effective for the treatment of vitamin D deficiency. Vitamin D level at birth and in early infancy has certain influence on neurobehavioral development.
Infant ; Child ; Infant, Newborn ; Humans ; Child, Preschool ; Cholecalciferol/pharmacology* ; Prospective Studies ; Fetal Blood ; Infant, Premature ; Dietary Supplements ; Vitamin D

This study was aimed to investigate the effect of 1,25(OH)(2) vitamin D(3) [1,25(OH)(2) Vit D(3)] on the differentiation, maturation and function of human dendritic cells (DC) in vitro and its mechanism. Human peripheral blood mononuclear cells were induced to differentiate to DC in vitro. The DC in test group were cultured with 1,25(OH)(2) Vit D(3) 1 nmol/L for 9 d, while the DC in control group were cultured with the equivalent of absolute alcohol. The expression of co-stimulatory molecules on DC were analyzed by flow cytometry. T cell proliferation induced by DC was assessed by MTT method. The expression of indoleamine 2, 3-dioxygenase (IDO) protein was determined by Western blot. The results showed that compared with the control group, the expression of CD80, CD83 and CD86 on DC in test group was significantly down-regulated (P < 0.05), while the CD1a was up-regulated (P < 0.05). The expression rate of CD80, CD83, CD86, CD1a in test group were (40.43 ± 9.83)%, (20.04 ± 4.73)%, (14.45 ± 5.38)%, (58.48 ± 10.72)% respectively, while in control group were (29.36 ± 13.34)%, (35.91 ± 10.19)%, (27.15 ± 11.64)%, (72.20 ± 12.79)% respectively. Compared with the control group, 1,25(OH)(2) Vit D(3)-treated DC exhibited a markedly reduced ability to stimulate allogenic T cell proliferation and up-regulated IDO protein expression.It is concluded that 1,25(OH)(2) Vit D(3) efficiently inhibits the maturation of DC and DC-mediated T cell proliferation, which may be related to the up-regulation of IDO protein expression.
Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cholecalciferol ; pharmacology ; Dendritic Cells ; cytology ; immunology ; Flow Cytometry ; Humans ; Immune Tolerance ; drug effects ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; metabolism

OBJECTIVETo observe the role of magnesium sulfate in vascular calcification, to explore the role and the mechanism of magnesium sulfate in vascular calcification.

METHODSThe vascular calcification model was established by administration of vitamin D3 plus nicotine (VDN) in SD rats. To estimate the extent of calcification by Von Kossa staining, calcium content and alkaline phosphatase activity, osteopontin (OPN) mRNA were determined by using semi-quantitative reverse-transcription polymerase chain reaction.The malondialdehyde (MDA) and nitric oxide (NO) content and activities of superoxide dismutase(SOD) were measured by biochemistry.

RESULTSA strong positive staining of black/brown areas among the elastic fibers of the medial layer in calcified aorta by Von Kossa staining, calcium content and ALP activity in calcified arteries increased by 3.9-and 3.4-fold as compared with the controls. The expression of OPN mRNA was up-regulated by 40% (P < 0.01). The lipid peroxidation products MDA in vascular were increased 2.0-fold (P < 0.01). The NO content and SOD activity were greatly decreased by 64% and 72% (P < 0.01), respectively, compared with controls. However, calcium content and ALP activity in VDN plus magnesium sulfate group were lower than those in VDN group. Low and high dosage magnesium sulfate obviously relieved degree of calcification in the cardiovascular tissues in a dosage-dependent manner (P < 0.01).

CONCLUSIONMagnesium sulfate plays a role in the pathogenesis of vascular calcification by reducing vascular calcification and decreasing vascular injury.

Animals ; Cholecalciferol ; adverse effects ; Magnesium ; pharmacology ; Male ; Nicotine ; adverse effects ; Osteopontin ; metabolism ; RNA, Messenger ; genetics ; Rats ; Vascular Calcification ; chemically induced ; pathology

Several chemical compounds can restore pigmentation in vitiligo through mechanisms that vary according to disease etiology. In the present study, we investigated the melanogenic activity of six structurally distinct compounds, namely, scopoletin, kaempferol, chrysin, vitamin D, piperine, and 6-benzylaminopurine. We determined their effectiveness, toxicity, and mechanism of action for stimulating pigmentation in B16F10 melanoma cells and in a zebrafish model. The melanogenic activity of 6-benzylaminopurine, the compound identified as the most potent, was further verified by measuring green fluorescent protein concentration in tyrp1 a: eGFP (tyrosinase-related protein 1) zebrafish and mitfa: eGFP (microphthalmia associated transcription factor) zebrafish and antioxidative activity. All the tested compounds were found to enhance melanogenesis responses both in vivo and in vitro at their respective optimal concentration by increasing melanin content and expression of TYR and MITF. 6-Benzyamino-purine showed the strongest re-pigmentation action at a concentration of 20 μmol·Lin vivo and 100 μmol·Lin vitro, and up-regulated the strong fluorescence expression of green fluorescent protein in tyrp1a: eGFP and mitfa: eGFP zebrafish in vitro. However, its relative anti-oxidative activity was found to be very low. Overall, our results indicated that 6-benzylaminopurine stimulated pigmentation through a direct mechanism, by increasing melanin content via positive regulation of tyrosinase activity in vitro, as well as up-regulating the expression of the green fluorescent protein in transgenic zebrafish in vivo.
Alkaloids ; chemistry ; pharmacology ; Animals ; Benzodioxoles ; chemistry ; pharmacology ; Benzyl Compounds ; chemistry ; pharmacology ; Cholecalciferol ; chemistry ; pharmacology ; Flavonoids ; chemistry ; pharmacology ; Humans ; Kaempferols ; chemistry ; pharmacology ; Melanins ; genetics ; metabolism ; Monophenol Monooxygenase ; genetics ; metabolism ; Pigmentation ; drug effects ; Piperidines ; chemistry ; pharmacology ; Polyunsaturated Alkamides ; chemistry ; pharmacology ; Purines ; chemistry ; pharmacology ; Scopoletin ; chemistry ; pharmacology ; Vitiligo ; drug therapy ; enzymology ; metabolism ; Zebrafish

OBJECTIVETo investigate the effect of hydrogen sulfide (H2S) on artherosclerosis (AS) and its mechanism in rats.

METHODS125 healthy male SD rats of the weight (210 +/- 10) g were randomly divided into 5 groups: control group, AS model group, AS + low-dose NaHS (2.8 micromol/(kg x d)) group, AS+ middle-dose (14 micromol/(kg x d)) NaHS group, AS+ high-dose NaHS (28 micromol/(kg x d)) group. The atherosclerotic model was established by feeding high grease food and injecting large doses of VitD3. The rats were using NaHS by peritoneal injection for 12 weeks. 5 rats were executed in each group before the experiment and in the weeks of 3, 6, 9, 12 after the experiment, respeotively. The blood fat was analyzed by automatic biochemistry analysator. H2S content in serum was detected by the method of deproteinization. The pathological damage of vessels was observed and scored by HE stain. The expression of VEGF in the vessel tissue was detected by immunohistochemistry staining.

RESULTSCompared with the control group at contemporaneity, both serumal triglyceride (TG) and cholesterol (TC) increased significantly in the AS model group after rat feeded 3, 6, 9, 12 weeks, and scores of the artery pathological damage also increased obviously from the 6th week to the 12th week (P < 0.01), as well as artherosclerosis plaque appeared, displaying as lipid plaque in the positive part. The serumal H2S concentration decreased obviously, from (44.98 +/- 2.06) micromol/L of before feeding to (38.56 +/- 2.26), (32.96 +/- 2.38), (28.63 +/- 0.92), (23.55 +/- 0.92) nnol/L of after feeding 3, 6, 9, and 12 weeks, respectively, and lower than that of control at contemporaneity (44.72 +/- 0.85), (43.71 +/- 0.59), (41.96 +/- 0.97), (39.87 +/- 1.25) micromol/L, respectively ( P < 0.01), and VEGF expression of the vascular tissue also increased (P < 0.01). Compared with the AS model group, all of above indexes in rat of the low-dose of NaHS group did not appear any obvious change. The serumal H2S concentration in rat of the middle-dose NaHS began increase at the 6 week after rat feeded (36.13 +/- 0.3 vs. 32.96 +/- 2.38 micronol/L, P < 0.05), and continuously increased at the 9th and the 12th week (33.07 +/- 1.14 vs. 28.63 +/- 0.92 micromol/L, 30.16 +/- 0.2 vs. 23.55 +/- 0.92 micromol/L; P < 0.01, respectively). The serumal H2S concentration in high-dose NaHS groups, increased from the 3th to the 12th week (41.25 +/- 0.80, 38.71 +/- 0.46, 35.31 +/- 0.62, 33.38 +/- 0.78 micromol/L, respectively, P < 0.01). The rat serumal TC in both middle and high-dose NaHS groups, decreased from the 3th to the 12th week (P < 0.01), and TG began decrease from the 3th and the 6th week to the 12th week after rat feeded, respectively (P < 0.05, P < 0.01). Both of the pathological damage scores and the expression of VEGF decrease from the 6th week to the 12th week (P < 0.05). The correlation analysis showed that H2S in serum had a negative correlation with both pathological damage scores (r = -0.917, P < 0.01) and the expression of VEGF (r = -0. 885, P < 0.01). But it had no obvious correlation with serumal TG and TC.

CONCLUSIONThe formation and development of artherosclerosis has a close correlation with the depressing of endogenous H2S. Administration of exogenous H2S could raise the H2S concentration of serum in artherosclerosis, which might improve the damage of vessels and inhibit the expression of VEGF.

Animals ; Atherosclerosis ; physiopathology ; prevention & control ; Cholecalciferol ; administration & dosage ; Dietary Fats ; administration & dosage ; Hydrogen Sulfide ; metabolism ; pharmacology ; therapeutic use ; Lipids ; blood ; Male ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

Differentiated HL-60 is an effector cell widely used for the opsonophagocytic-killing assay (OPKA) to measure efficacy of pneumococcal vaccines. We investigated the correlation between phenotypic expression of immunoreceptors and phagocytic ability of HL-60 cells differentiated with N,N-dimethylformamide (DMF), all-trans retinoic acid (ATRA), or 1alpha, 25-dihydroxyvitamin D3 (VitD3) for 5 days. Phenotypic change was examined by flow cytometry with specific antibodies to CD11c, CD14, CD18, CD32, and CD64. Apoptosis was determined by flow cytometry using 7-aminoactinomycin D. Function was evaluated by a standard OPKA against serotype 19F and chemiluminescence-based respiratory burst assay. The expression of CD11c and CD14 gradually increased upon exposure to all three agents, while CD14 expression increased abruptly after VitD3. The expression of CD18, CD32, and CD64 increased during differentiation with all three agents. Apoptosis remained less than 10% until day 3 but increased after differentiation by DMF or ATRA. Differentiation with ATRA or VitD3 increased the respiratory burst after day 4. DMF differentiation showed a high OPKA titer at day 1 which sustained thereafter while ATRA or VitD3-differentiated cells gradually increased. Pearson analysis between the phenotypic changes and OPKA titers suggests that CD11c might be a useful differentiation marker for HL-60 cells for use in pneumococcal OPKA.
Antibodies, Bacterial/immunology ; Antigens, CD11c/metabolism ; Antigens, CD14/metabolism ; Antigens, CD18/metabolism ; Apoptosis/*immunology ; Biological Assay ; Cell Differentiation ; Cell Line, Tumor ; Cholecalciferol/pharmacology ; Dimethylformamide/pharmacology ; Flow Cytometry ; HL-60 Cells ; Humans ; Phagocytosis/*immunology ; Pneumococcal Vaccines/*immunology ; Receptors, IgG/metabolism ; Receptors, Immunologic/*biosynthesis ; Respiratory Burst/immunology ; Streptococcus pneumoniae/*immunology ; Tretinoin/pharmacology

Differentiated HL-60 is an effector cell widely used for the opsonophagocytic-killing assay (OPKA) to measure efficacy of pneumococcal vaccines. We investigated the correlation between phenotypic expression of immunoreceptors and phagocytic ability of HL-60 cells differentiated with N,N-dimethylformamide (DMF), all-trans retinoic acid (ATRA), or 1alpha, 25-dihydroxyvitamin D3 (VitD3) for 5 days. Phenotypic change was examined by flow cytometry with specific antibodies to CD11c, CD14, CD18, CD32, and CD64. Apoptosis was determined by flow cytometry using 7-aminoactinomycin D. Function was evaluated by a standard OPKA against serotype 19F and chemiluminescence-based respiratory burst assay. The expression of CD11c and CD14 gradually increased upon exposure to all three agents, while CD14 expression increased abruptly after VitD3. The expression of CD18, CD32, and CD64 increased during differentiation with all three agents. Apoptosis remained less than 10% until day 3 but increased after differentiation by DMF or ATRA. Differentiation with ATRA or VitD3 increased the respiratory burst after day 4. DMF differentiation showed a high OPKA titer at day 1 which sustained thereafter while ATRA or VitD3-differentiated cells gradually increased. Pearson analysis between the phenotypic changes and OPKA titers suggests that CD11c might be a useful differentiation marker for HL-60 cells for use in pneumococcal OPKA.
Antibodies, Bacterial/immunology ; Antigens, CD11c/metabolism ; Antigens, CD14/metabolism ; Antigens, CD18/metabolism ; Apoptosis/*immunology ; Biological Assay ; Cell Differentiation ; Cell Line, Tumor ; Cholecalciferol/pharmacology ; Dimethylformamide/pharmacology ; Flow Cytometry ; HL-60 Cells ; Humans ; Phagocytosis/*immunology ; Pneumococcal Vaccines/*immunology ; Receptors, IgG/metabolism ; Receptors, Immunologic/*biosynthesis ; Respiratory Burst/immunology ; Streptococcus pneumoniae/*immunology ; Tretinoin/pharmacology

OBJECTIVETo investigate the effects of Chinese kidney-tonifying drugs on bone mineral density, biomechanics, 25-hydroxy Vitamin D3 and 1,25-dihydroxy Vitamin D3 of ovariectomized osteoporosis rats, and explore the mechanism of treating osteoporosis with the drugs.

METHODSThirty-six female SD rats (four months) were randomly divided into model group, sham group and treatment group. All the rats had been ovariectomied except those in sham group. Selecting 4, 8, 12 weeks in the experiment, the value of bone mineral density (BMD) was measure by dual energy X-ray absorptiometry (DEXA) of femoral head, while the biomechanics machine was applied to analysis femoral head biomechanics index and ELISA method was used to detect the content of 25-hydroxy Vitamin D3 and 1,25-dihydroxy Vitamin D3 discern in blood-serum, liver and kidney.

RESULTSTreatment group rats' BMD of femoral head was enhance compared with model group, significant differences were absent (P<0.05), and the maximal load and maximal stress measurement were improved, significant differences were absent (P<0.05). As the content of 25-hydroxy Vitamin D3 and 1,25-dihydroxy Vitamin D3 discern in blood-serum, liver and kidney were elevate, furthmore there were significant differences in group comparison, all significant differences were absent (P<0.05). But those compared with sham group, there was no significant difference (P>0.05).

CONCLUSIONIn the early period in absence of estrogenic hormone, the Chinese kidney-tonifying drugs could activate bone metabolism to raise BMD and reinforce quality of bone through up-regulating expression of 25-hydroxy Vitamin D3 and 1,25-dihydroxy Vitamin D3 at protein level.

Animals ; Biomechanical Phenomena ; drug effects ; Bone Density ; drug effects ; Cholecalciferol ; analogs & derivatives ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Femur Head ; drug effects ; metabolism ; Humans ; Osteoporosis ; drug therapy ; metabolism ; physiopathology ; Ovariectomy ; adverse effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Renal Agents ; pharmacology