Effects of ghrelin on the proliferation and differentiation of 3T3-L1 preadipocytes and its possible mechanisms / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the effects of ghrelin on the proliferation and differentiation of 3T3-L1 preadipocyte, and study the possible mechanisms.</p><p><b>METHODS</b>3T3-L1 preadipocytes were cultured in vitro. The proliferation potentials of 3T3-L1 preadipocytes that were treated with different concentrations of ghrelin were evaluated by MTT methods. The levels of c-myc and thymidine kinase mRNA were detected using RT-PCR. 3T3-L1 preadipocytes were differentiated into the matured adipocytes with insulin (INS) or ghrelin. The morphological changes of 3T3-L1 adipocytes were observed and the differentiation rate was assayed by oil-red O staining. Total RNA was extracted from adipocytes at various times, and the levels of peroxisome proliferation activated receptor gamma (PPARgamma) and CAAT/enhancer binding protein(C/EBPalpha) mRNA expressions were detected using RT-PCR.</p><p><b>RESULTS</b>Ghrelin at concentrations of 10(-7) to 10(-15) mol/L significantly stimulated preadipocyte proliferation (p<0.05). The levels of c-myc and thymidine kinase mRNA significantly increased in 3T3-L1 preadipocytes with 10(-9) mol/L and 10(-11) mol/L ghrelin treatment (p<0.01). The 3T3-L1 preadipocytes treated with 10(-11) mol/L ghrelin had lots of lipid droplets in the cytoplasma, but the differentiation rate was lower than those treated with INS. Ghrelin of 10(-11) mol/L significantly increased the mRNA expression of PPARgamma and C/EBPalpha in the course of 3T3-L1 preadipocyte differentiation, compared with the normal control group (p<0.05). The PPARgamma and C/EBPalpha mRNA expression increased with the prolonged differentiation of preadipocytes induced by ghrelin or INS. There were significant differences in the levels of PPARgamma and C/EBPalpha mRNA expression between the 2nd and 8th days of differentiation(p<0.01).</p><p><b>CONCLUSIONS</b>Ghrelin promotes the proliferation and differentiation of 3T3-L1 preadipocytes. The proliferation of 3T3-L1 preadipocytes induced by ghrelin may be associated with increased c-myc levels. Ghrelin may promote differentiation of 3T3-L1 preadipocytes by increasing mRNA expression of PPARgamma and C/EBPalpha, thus enhances the sensitivity of adipocytes to INS.</p>

Effect of acylation stimulating protein on the perilipin and adipophilin expression during 3T3-L1 preadipocyte differentiation / 生理学报

Perilipin and adipophilin, two significant lipid droplet (LD)-specific proteins, participate in storing fat or ectopic lipid deposition and fat mobilization in many types of mammalian cells. Acylation stimulating protein (ASP) is a novel adipocyte-derived hormone known for a major determinant for triglyceride synthesis (TGS) and lipid metabolism. The present study was aimed to investigate: (1) whether ASP, rather than insulin, is a powerful potentiator which could physiologically and directly influence TGS during 3T3-L1 preadipocyte differentiation; (2) whether ASP exposure at indicated time points during 3T3-L1 preadipocyte differentiation could influence the gene/protein expression of adipophilin and perilipin. 3T3-L1 preadipocytes were differentiated by traditional hormone cocktail and divided into control, ASP and insulin groups according to the treatment of ASP (1 mmol/L) or insulin (100 nmol/L). ASP-stimulated and insulin-stimulated TGS rate at indicated time points (0 d, 3 d, 6 d, 9 d) were assayed by measuring the incorporation of [(3)H]-oleic acid into TG, and the corresponding glucose transport was assayed by [(3)H]-2-DG uptake. The effects of ASP or insulin on gene/protein expression of adipophilin and perilipin at indicated time points were evaluated by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The results obtained were as follows: (1) on the 3rd and 6th day of differentiation, ASP dramatically enhanced TGS rate compared with control group (P<0.05, P<0.01); There was no significant difference in TGS rate between insulin group and control group; (2) on the 6th and 9th day of differentiation, both ASP and insulin promoted glucose uptake (P<0.05, P<0.01), and the promoting effect in ASP group was greater than that in insulin group; (3) ASP elevated adipophilin gene and protein expression at the very early stage of differentiation (P<0.05, P<0.001) and had no significant effect from the 4th day of differentiation. Perilipin gene and protein expression increased throughout preadipocyte differentiation and its expression was up-regulated following ASP stimulation from the 3rd day of differentiation (P<0.05, P<0.001) to the end of differentiation (P<0.05); (4) Insulin did not affect gene and protein variation pattern of adipophilin and perilipin. Taken together, this study provides evidence that ASP-evoked changes in gene and protein expression of adipophilin and perilipin correlate with ASP-stimulated TGS acceleration, and adipophilin and perilipin are involved in the molecular mechanism of ASP-induced adipogenesis and LD formation.

Clinical characteristics of Wolfram syndrome / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Wolfram syndrome (WFS) is a rare, autosomal recessive inherited disease characterized by various clinical manifestations. The aim of this study was to investigate clinical characteristics of WFS.</p><p><b>METHODS</b>One case of WFS was reported. Combined with the clinical data of 8 cases of WFS which had been reported in China between 1994 and 2007, the clinical characteristics of WFS were reviewed.</p><p><b>RESULTS</b>Insulin-dependent diabetes mellitus as the earliest manifestation was found in all of the 9 patients, with a median onset age of 5.0 years. Optic atrophy occurred in 8 patients (onset age: 8.5 years), diabetes insipidus in 7 patients (onset age: 8.5 years) and deafness in 7 patients (onset age: 9.8 years). Short stature was found in 6 patients and hydroureteronephrosis in 4 patients.</p><p><b>CONCLUSIONS</b>Insulin-dependent diabetes mellitus was the first presentation in children with WFS. Optic atrophy, diabetes insipidus and deafness were common complications, with a various onset age.</p>

Childhood gynecomastia: a clinical analysis of 240 cases / 中国当代儿科杂志

Two hundred forty cases of childhood gynecomastia were studied retrospectively. There were 13 cases aged 3 to 10 years and 227 cases aged 11 to 15 years. Of the 240 cases of gynecomastia, 160 presented with bilateral breast enlargement, 50, left breast enlargement, and 30, right breast enlargement. The etiology of gynecomastia of the 240 patients included adolescent breast hyperplasia (n=219), drug ingestion (n=2), and secondary causes (n=5). Fourteen patients did not show identifiable causes and were diagnosed as idiopathic gynecomastia. The 8 patients with identifiable causes received specific treatment. After 1-3 months of treatment, the breasts of the patients improved. The 219 cases of adolescent breast hyperplasia and 14 cases of idiopathic gynecomastia were not given any medication. They were followed up regularly. Most of the patients recovered well within a 27-month follow-up.

Expression of Daxx in children with acute leukemia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate Daxx expression and its clinical significance in children with acute leukemia.</p><p><b>METHODS</b>The expression of Daxx protein was detected by immunohistochemical assay in 50 children with newly diagnosed acute leukemia (34 cases of acute lymphocytic leukemia and 16 cases of acute non-lymphocytic leukemia). Twenty children with normal bone marrow were used as the control group.</p><p><b>RESULTS</b>Daxx protein was expressed in 38.0% of 50 children with acute leukemia, which was significantly higher than that of the control group (5.0%) (P < 0.05). The children with acute non-lymphocytic leukemia had significantly higher Daxx expression levels (62.5%) than those with acute lymphocytic leukemia (26.5%; P < 0.05) as well as the control group (P < 0.05). There were no significant differences in the Daxx expression between acute lymphocytic leukemia children and the control group. Daxx protein was expressed in 55.6% of high risk group of acute lymphocytic leukemia but it was not expressed in standard risk group of acute lymphocytic leukemia (P < 0.05).</p><p><b>CONCLUSIONS</b>Daxx expression is abnormal in children with acute leukemia and associated with some clinical features of acute leukemia, suggesting that it may play an important role in the genesis and development of acute leukemia.</p>

Expression of Kiss-1 mRNA in the hypothalamus of true precocious female rats / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the gene expression of Kiss-1 in the hypothalamus of true precocious female rats at various stages of development.</p><p><b>METHODS</b>Forty 5-day-old normal female Sprague-Dawley rats were randomly assigned into four groups of 10 rats: Control group 1, Control group 2, Model group 1 and Model group 2. The rats from the two model groups were injected with 300 microg of danazol at 5 days of age to induce true precocious puberty. The two control groups were injected with normal saline instead. For the determination of Kiss-1 mRNA expression in the hypothalamus, the rats of the Model group 1 were sacrificed during the first diestrus (early puberty) and meanwhile the rats of the Control group 1 were sacrificed when they were at prepuberty; the Control group 2 rats were sacrificed at the first diestrus (early puberty); the rats of the Model group 2 were sacrificed during the second diestrus (middle puberty). The expression of Kiss-1 mRNA in the hypothalamus in the four groups was detected using semi-quantitative reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>Kiss-1 mRNA expression in the hypothalamus in Model group 1 and Model group 2 increased by 1.4-fold and 2.8-fold, respectively, compared with that of Control group 1 (P < 0.05). Model group 2 showed significantly higher Kiss-1 mRNA levels than Model group 1 (P < 0.05). There were no statistical differences in the Kiss-1 mRNA expression between Control group 2 and Model group 1.</p><p><b>CONCLUSIONS</b>Gene expression of Kiss-1 is associated with the developmental period of true precocious puberty, suggesting that Kiss-1 might play a role in the pathogenesis of this disorder.</p>

Effect of Oleic Acid on Differentiation of SW872 Preadipocytes / 实用儿科临床杂志

Objective To study the effect of oleic acid on the differentiation of SW872 preadipocytes.Methods SW872 prea-(dipocytes) were cultured and induced to differentiate by 0.6 mmol/L oleic acid in vitro.After 24 h,48 h and 72 h of differentiation,the morphological changes of SW872 preadipocytes were observed and the differentiation rate was assayed by oil-red O staining.In addition,triglyceride(TG) mass was detected by chemical colorimetry methods.During the differentiation of SW872 preadipocytes,transcription factors including peroxisome proliferator activated receptor-?_2(PPAR-?_2) and CAAT/enhancer binding protein-?(C/EBP-?)(mRNA) were also measured by reverse transcription-polymerase chain reaction(RT-PCR).Results 1.SW872 preadipocytes were fibroblastic and had no obvious fat droplet in cytoplasm.However,when stimulated for 72 hby 0.6 mmol/L oleic acid,SW872 preadipocytes became more bigger and rounder and differentiated into mature adipocytes with lots of fat droplets in the cells.2.Compared with that of predifferentiation,the concentration of TG mass increased by 14 folds after 72-hour differentiation(P

Clinical Significance of Detection of Serum Interleukin-8 in Children with Thyroid Disfunction / 实用儿科临床杂志

Objective To investigate the changes of serum interleukin 8(IL-8) level in children with thyroid disfunction and its clinical significance in children with autoimmune thyroid disease(AITD).Method Serum IL-8 of 30 children with hyperthyroidism,16 children with hypotyroidism and 20 health children were detected by double antibody enzyme linked immunossorbent assay(ELISA).Results Serum IL-8 levels in hyperthyroidism and hypothyroidism children were significantly higher than that in controls(P0.05).With the remission of the diseases, IL-8 levels decreased; there was a significant difference between serum IL-8 levels in patients before and after therapy (P

Gene Expression of Adiponectin Receptors during Diffe rentiation of SW872 Preadipocytes / 实用儿科临床杂志

Objective To explore genes expression of adiponectin receptors during differentiation of SW872 preadipocytes. Met-(hods) SW872 preadipocytes were cultured in vitro and induced to differentiate by 0.6 mmol/L oleic acid. During the progress of diffe-(rentiation), the morphological changes of SW872 cells were observed and the differentiation rate was assayed by oil-red O staining. Adiponectin receptors mRNA was measured by reverse transcription-polymerase chain reaction during differentiation of SW872 preadipocytes. Results 1.After stimulated by 0.6 mmol/L oleic acid for 72 hours, almost all SW872 cells were differentiated,and there were lots of fat droplets in the cells.2.There were adiponectin receptors genes expressions in SW872 preadipocytes.After 72 hours,and the levels of adiponectin receptor(AdipoR) 1 mRNA and AdipoR 2 mRNA were markedly increased up to 2.54 and 4.09 times,respectively. Conclusion There are AdipoR1 and AdipoR2 genes expressions in fat cells and the expressions are differentiation-dependent.

Application of gas chromatography-mass spectrometry analysis on urine filter paper in the high-risk screening and diagnosis of inherited metabolic diseases / 中华儿科杂志

<p><b>OBJECTIVE</b>To establish a specific procedure for the high-risk screening and diagnosis of organic acidurias and other inherited metabolic diseases in China.</p><p><b>METHODS</b>A nation-wide network for the high-risk screening and diagnosis of genetic metabolic diseases was formed to facilitate the collaboration. Urine samples were collected using filter paper from patients with clinical symptoms suspicious of inherited metabolic diseases. The samples were eluted with distilled water and internal standards were added. Samples were treated with hydroxylamine hydrochloride to form oximes to improve the recoveries of 2-ketoacids. Urinary organic acids were extracted with ethyl acetate and diethyl ether under acidic condition. After dehydration, the combined organic phase was evaporated to dryness with nitrogen. The residues were added with BSTFA + 1%TMCS and heat incubated to form the trimethylsilyl derivatives, and then were analyzed on an Agilent 5890/5973N gas chromatography-mass spectrometer (GC-MS), with a 7683 series auto-sampler. The peaks were identified by reference to a mass spectral library.</p><p><b>RESULTS</b>Totally 352 samples were collected from the network collaborating hospitals since 2001. Thirty-four (9.66%) cases of various inherited metabolic diseases were diagnosed with an age range of 2 days to 14 years. The disease profile was consisted of methylmalonic acidemias (6), alpha-keto-glutaric aciduria (5), tyrosinemia type I (4), dicarboxylic aciduria (4), multiple carboxylase deficiency (3), phenylketonuria (3), lactic acidemia (3), propionic acidemia (2), ornithine transcarbamoylase deficiency (1), ethylmalonic-adipic aciduria (1), glutaric aciduria type II (1) and 3-methylcrotyl CoA carboxylase deficiency (1). The most common clinical symptoms and signs included mental and developmental retardation, convulsion, musculotonic abnormality and jaundice. Routine laboratory tests often revealed metabolic acidosis, hypoglycemia and hyperammonemia, etc.</p><p><b>CONCLUSION</b>Urine organic acids analysis by GC-MS remains to be the most important technique for the high-risk screening and diagnosis of inherited metabolic diseases. Use of urine filter paper for sample collection and analysis in advanced genetic metabolic centers is a practical approach to extend the diagnostic capacity and improve the management of such diseases in China. Collaborative network played a critical role in the success of the program.</p>

Recombinant human interleukin-6 inhibits mRNA expressions of adiponectin,its receptor 1 and adiponectin secretion in SW872 adipocytes / 中华内分泌代谢杂志

SW872 preadipocytes were cultured and induced to differentiate by oleic acid in vitro.The levels of adiponectin and its receptors (AdipoR1 and AdipoR2) mRNA were measured by semiquantitative RT- PCR.The concentration of adiponectin in the culture medium was assayed by ELISA.The results showed that rhIL- 6 could inhibit adiponectin,AdipoR1 mRNA expressions and adiponectin secretion in SW872 adipocytes in a time- and dose-dependent manner,but did not influence adipoR2 mRNA expression.

Influence of Thyroid Status on Serum Levels of Leptin and Leptin mRNA in Rats / 实用儿科临床杂志

Objective To explore the influence of thyroid status on serum level of leptin and expression of leptin mRNA in rats with hypothyroidism or hyperthyroidism.Methods Fifty rats were artificially rendered hyperthyroidism and hypothyroidism by L-thyroxine or methimazole and divided into 5 groups.The serum of T 3,T 4,TSH,leptin concentration and expression of leptin mRNA were measured in 5 groups.Results Serum leptin concentration was significantly correlated with weight of rats in 5 groups(P

Effect of recombinant human interleukin-6 on the secretion of acylation-stimulating protein in SW872 adipocytes / 中华内分泌代谢杂志

SW872 cells were cultured in vitro with oleic acid and differentiated into mature adipocytes. The role of interleukin-6(IL-6)in the secretion of acylation-stimulating protein(ASP)in mature SW872 adipocytes was observed.The results suggested that IL-6 significantly inhibited ASP secretion into the media in a dose-and time-dependent manner in mature SW872 adipocytes.

Changes of gene expressions of adiponectin in adipose tissue and adiponectin receptor R1 in skeletal muscle of type 2 diabetic rats / 中华内分泌代谢杂志

The mRNA levels of adiponectin in adipose tissue and adiponectin receptor R1 in skeletal muscle of type 2 diabetic rats were examined by semi-quatitative RT-PCR.The expression of adiponectin receptor R1 was not altered in the skeletal muscle of type 2 diabetic rats as compared with normal rats.The serum adiponectin level was decreased in diabetic rats due to the decline of adiponectin mRNA level in adipose tissue and rosiglitazone improved the adiponectin deficiency.