Purpose: The present study aimed to compare the efficacy and safety of recombinant human growth hormone (rhGH) therapy between children with idiopathic short stature (ISS) and growth hormone deficiency (GHD). Methods: A total of 150 pediatric patients with ISS and 153 pediatric patients with GHD who received rhGH treatment for more than one year from 2005 to 2016 were enrolled. Growth velocity (GV); height standard deviation (HtSD); insulin-like growth factor-1 standard deviation (IGF-1SD); body mass index (BMI); and the incidence of fasting hyperglycemia, fasting hyperinsulinemia, and hypothyroidism were recorded and compared. Results: At the beginning of treatment, chronological age, bone age, height, and BMI were not statistically significant between the two groups. rhGH dosage in ISS was significantly higher compared with GHD (P = 0). GV from half a year to three years after rhGH therapy was higher in the GHD group compared with the ISS group, but the differences were not statistically significant (P > 0 .05). HtSD increased in the two groups after rhGH therapy. HtSD at the beginning and after three years of therapy was not different between groups except for after half a year of therapy. HtSD in patients with ISS was significantly higher compared with GHD (P < 0 .05). The incidence of hypothyroidism was significantly higher in the GHD group compared with the ISS group (13.72% vs. 6.0%; P < 0.05). Moreover, the incidence of hyperinsulinemia was significantly higher in the ISS group compared with the GHD group (15.33% vs. 7.84%; P < 0 .05). Conclusions: rhGH increases growth in children with ISS and GHD. Fasting insulin and thyroid function were closely monitored for long-term follow up.

Objective:To evaluate the effects of visfatin on the protein expression of insulin signal molecules including insulin receptor substrate-1(IRS-1),insulin receptor substrate-2(IRS-2) and phosphatidylinositol 3-kinase(PI3K) on the states of insulin resistant in SW872 Adipocytes.Methods:Pre-adipocytes of the line SW872 were cultured and induced to differentiate into matured SW872 adipocytes.Then the cells were treated with oleate at concentration of 1.0mmol/L for 24hto induce insulin resistance.And the cells were cultured with Visfatin at concentration of 100nmol/L for 1h,and then the cell proteins were extracted.Western blot assay was used to detect the protein expression of IRS-1,IRS-2,and PI3K.Results:In the normal states,compared with the control group,100nmol/L visfatin promoted the protein expression of IRS-1,IRS-2,and PI3Kin SW872 adipocytes significantly.The levels of IRS-1,IRS-2,and PI3K were increased respectively by 51.65%(P0.01),44.74%(P0.01),and 61.38%(P0.01).In the insulin resistant states,after the stimulating by visfatin,the protein expression of IRS-1,IRS-2and PI3Kwere increased by 26.98%(P0.05),35.59%(P0.05),and 27.61%(P0.01).Conclusion:Visfatin might promote the glucose transport and play aphysiological role in the insulin resistance in SW872 adipocytes by modulating the signaling molecules of IRS-1,IRS-2,and PI3K.

To explore the relationship between visfatin, ASP and the incidence of childhood obesity and the significance of them in diagnoses and therapy of children obesity. Eighty-six children (57 boys and 29 girls) including 40 obese children,22 overweight children and 24 normal control children were recruited with ages ranged 7-15 years. Serum visfatin and ASP levels were determined by ELISA. (1) As compared with normal and overweight children, serum visfatin level was significantly higher in obese children (p<0.05);As compared with normal children (p<0.01)and overweight children (p<0.05), serum ASP level was significantly higher in obese children. There were no differences of serum visfatin and ASP levels between normal and overweight children. (2) The body mass index, TC, TG, LDL-c, FPG, FINS, insulin resistance index of the obese children were higher than the normal children (p<0.05 or p<0.01); the HDL-c and insulin sensitivity index of the obese children were lower than the normal group (p<0.01 or p<0.05). The body mass index, the fasting insulin, insulin resistance index of the obese children were higher than the overweight children (p<0.01). (3) Correlation Analysis: Serum visfatin and body mass index and TG were positively correlated (respectively r = 0.218, p<0.05; r = 0.500, p<0.01). ASP and body mass index, total cholesterol, triglycerides were positively correlated (respectively r = 0.268, p<0.05; r = 0.250, p <0.05; r = 0.427, p<0.01). In simple obesity children, there were a significant change in serum visfatin and ASP, and visfatin and ASP involved in the disorder of lipids metabolism in obese children. Serum visfatin and ASP levels can be used as a new indicator to evaluate the risk of childhood obesity trends and future valuation of diabetes, cardiovascular.

Summary. The chemokine monokine induced by interferon‐γ (Mig) is involved in the recruitment of inflammatory cells and liver injury during hepatitis B virus (HBV) infection. HBV protein X contributes to Mig expression in vitro by activation of nuclear factor (NF)‐κB; however, the molecular mechanisms by which HBV induces Mig expression in vivo are unknown. In this paper, we established a mouse model for HBV study by tail vein injection of HBV genome‐containing adenovirus vectors. Host immune response to the secreted hepatitis B surface antigen and e antigen was detected and serum alanine aminotransferase (ALT) was elevated at different time points. We also demonstrated that peripheral and intrahepatic Mig expression was increased after Ad‐HBV infection. This was followed by inflammatory cell migration and formation of inflammatory foci in the liver. In addition, NF‐κB p65 subunit translocated from the cytoplasm to the nucleus, and phosphoinositide 3‐kinase/Akt, extracellular signal‐regulated kinase (ERK) and c‐Jun N‐terminal kinase (JNK) were to some extent phosphorylated after HBV injection. Following tail vein injection of Mig siRNA/in vivo‐jetPEI‐Gal complex, Mig expression was partially suppressed, inflammatory cell migration was inhibited, serum level of ALT were reduced. In conclusion, through NF‐κB activation, HBV induced Mig expression in vivo, which recruited peripheral inflammatory cells to the liver and resulted in liver damage. Phosphorylation of phosphoinositide 3‐kinase/Akt, ERK and JNK but not p38 might involved in the molecular mechanisms underlying HBV induced Mig expression in vivo.

The novel adipokine acylation stimulating protein (ASP) is involved in lipid metabolism and obesity-related disorders. Adipophilin and perilipin, two members of the lipid droplet protein family, participate not only in fat storage within adipocytes, but also in ectopic lipid deposition in the form of cytoplasmic triglyceride (TG) droplets within many types of mammalian cells. During differentiation to mature adipocytes, mechanisms controlling the synthesis and turnover of these lipid droplet proteins are only partially understood, the mechanisms regulating gene/protein expression as yet unidentified. In our previous study, ASP has been shown to regulate adipophilin and perilipin expression to facilitate TG synthesis during 3T3-L1 cell differentiation. Our aim in this study was to provide insight into the physiological importance of phosphoinositide 3-kinase (PI3K) and phospholipase C (PLC) in ASP-triggered alteration of adipophilin and perilipin expression. We found that acute (2.5 h) inhibition of PLC or PI3K results in a decrease in mRNA and protein of perilipin and adipophilin at any time during differentiation. The fact that there is such a rapid change even with mRNA levels suggests a rapid turnover of both mRNA and protein independent of a direct ASP effect. Also, the presence of these inhibitors blocked the ASP stimulatory effects with a maximal decrease in gene and protein expression of adipophilin (-45% and -60%, respectively, P < 0.01) and perilipin (-96% and -63%, respectively, P < 0.01 and P < 0.05). These findings provide further understanding of the adipogenic properties of ASP in adipocytes.

Objective To explore the roles of Visfatin on insulin resistance(IR) in cultured SW872 matured adipocytes induced by oleic acid.Methods SW872 preadipocytes were cultured in vitro and induced to differentiated by 0.6 mmol·L-1 oleic acid for 48 h.Then the SW872 preadipocytes were differentiated into the matured adipocytes.There were 2 group in the study,normal control group:normal SW872 matured adipocytes;IR group:the normal SW872 matured adipocytes were induced by 1.0 mmol·L-1 oleic acid for 24 h.Glucose transport was assessed by[3H]-2-deoxy glucose uptake.Results 1.Glucose transport stimulated by Visfatin and Insulin in normal control group were significantly increased by 110% (P0.01) and 134% (P0.01) compared with basis state group,respectively.2.In IR group,basic glucose transport was reduced by 9.73% (P0.05),glucose transport stimulated by Visfatin was reduced by 12.30 % (P0.01) compared with normal control group.Glucose transport stimulated by Insulin was reduced by 26.81 % (P0.01),basic glucose transport was reduced by 10.50% (P0.05) compared with normal control group.3.In IR group,glucose transport stimulated by Insulin was not increased significantly (P0.05) compared with the same basis state group;Glucose transport stimulated by Visfatin was significantly increased by 6.9%(P0.05) compared with the same basis state group.Conclusion Visfatin can promote glucose transport and improve IR in SW872 matured adipocytes.

OBJECTIVE To investigate the effects of ghrelin on the proliferation and differentiation of 3T3-L1 preadipocyte, and study the possible mechanisms. METHODS 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. RESULTS 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). CONCLUSIONS 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.

OBJECTIVE 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. METHODS 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. RESULTS 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. CONCLUSIONS 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.

AIM: To investigate the role of individual protein isoforms in the regulation of 3T3-L1 adipocyte development through detecting temporal patterns of Gβ,Gα/q,phosphorylated phosphoinositide 3-kinase IA and IB isoforms and activated protein kinase C α and ζ subtypes involved in Gβ/Gαq-PI3K-PKC signaling pathway during 3T3-L1 preadipocyte differentiation.METHODS: The cells were induced by 1-methyl-3-isobutylxanthine,dexamethasone and insulin in vitro,and harvested at indicated time points(0 d,6 h,12 h,1 d,3 d,6 d,9 d),then the total proteins of these cells were extracted.The expressions of Gβ,Gα/q,p101,phosphorylated p85,p55,p110γ,PKCα and ζ were assayed by Western blotting.RESULTS: (1) Gαq/11 and Gβ increased after induction of differentiation,reaching maxima respectively at 3 d and 1 d when cellular level of Gβ was 1.97±0.16-fold higher and expression of Gαq/11 was 2.34±0.22-fold higher than that in just-confluent(0 d) cells.Subsequently the expression declined.(2) Compared to 0 d,phosphorylated p110γ,p55 and p85 elevated slightly at 12 h,and decreased significantly by the end of the treatment period(9 d) which coincided with maximal differentiation.While the expression of p101 elevated slightly at 12 h,no statistical significance through differentiation was found.(3) Phosphorylated PKCα increased significantly,peaking at 3 d of differentiation.Expression of phosphorylated PKCζ decreased during differentiation and its level 45.52% lower in adipocytes than that in preadipocyte was observed.CONCLUSION: The protein levels elevating at the early stage of differentiation correlate with the time point at which clonal expansion of cells is observed.Phosphorylated p55 and PKCζ decrease in adipocytes than that in preadipocyte,indicating an inhibitory influence upon the late differentiation process.

脑肠肽是一种胃源性促生长激索释放多肽,是生长激素促分泌素受体的内源性配体.作为一种与生长和能量代谢密切相关的激素,脑肠肽在体内起到调控能量正平衡、促进生长的作用.脑肠肽与其受体结合后能促进生长激素的释放,并且其促进生长激素释放的作用明显大于促生长激素释放激素,进而在儿童生长发育过程中发挥重要作用.脑肠肽在发挥该作用的同时也受到其他因素的调控,其作用机制和调节机制较为复杂.目前有关脑肠肽与生长落后关系的研究尚存在争议。

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.

The present study evaluated the application of three dimensional echocardigraphy (3DE) in the diagnosis of atrial septal defect (ASD) and the measurement of its size by 3DE and compared the size with surgical findings. Two-dimensional and real-time three dimensional echocardiography (RT3DE) was performed in 26 patients with atrial septal defect, and the echocardiographic data were compared with the surgical findings. Significant correlation was found between defect diameter by RT3DE and that measured during surgery (r=0.77, P<0.001). The defect area changed significantly during cardiac cycle. Percentage change in defect size during cardiac cycle ranged from 6%–70%. Our study showed that the size and morphology of atrial septal defect obtained with RT3DE correlate well with surgical findings. Therefore, RT3DE is a feasible and accurate non-invasive imaging tool for assessment of atrial septal size and dynamic changes.

Objective To explore the effect of different concentrations of recombinant human visfatin on glucose transport in cultured SW872 adipocytes.Methods SW872 preadipocytes were cultured in vitro and induced to differentiate by 0.6 mmol/L oleic acid for 48 h.During the progress of differentiation,the morphological changes of SW872 cells were observed by optics microscope.When SW872 preadipocytes were differentiated into the matured adipocytes,visfatin was added into the culture medium of fully differentiated adipocytes for 24 h.Following the exposure of adipocytes to visfatin at different concentrations,glucose transport was measured by 2-deoxy-[3H]-dexiontropic glucose incorporation method.Results SW872 readipocytes were fibroblastic and had no obvious fat droplet in cytoplasm.However,when stimulated for 72 h by 0.6 mmol/L oleic acid,SW872 preadipocytes became even bigger,and rounder,and differentiated into mature adipocytes with lots of fat droplets in the cells.SW872 adipocytes incubated for 24 h with different concentrations of visfatin showed that,under ba-sal states and insulin stimulation,5 nmol/L and 10 nmol/L visfatin had no effect on the glucose uptake.But 25,50,100 and 200 nmol/L visfatin significantly increased glucose uptake of basal states by 2.90%,7.70%,12.01% and 12.29% compared with control groups,and that of insulin stimulation states by 3.40%,16.37%,30.30% and 30.83% compared with control groups,respectively(Pa0.05,0.01).The glucose uptake of basal states and insulin stimulation states reached an approaching maximal effect at 100 nmol/L visfatin in SW872 adipocytes,showed significantly different compared with 50 nmo/L visfatin and control group(Pa0.01).There were no significant differences between 200 nmol/L and 100 nmo/L visfatin group(Pa0.05).Conclusion Visfatin increases glucose transport in SW872 adipocytes in concentration-dependent manner,after exposure to 100 nmol/L visfatin,the glucose uptake reaches an approaching maximal effect.

近几年,因乳房发育而就诊的男性儿童及青少年有逐渐增多的趋势,国内外就男性儿童乳房发育的病因、诊断和治疗也时见报道[1-6].为总结该病的诊疗经验,笔者对本院2004年1月～2007年1月内分泌门诊就诊的21例男性儿童及青少年非生理性乳房发育的资料,进行回顾性分析,现报告如下.

原核生物、真核生物、植物体的非神经细胞和组织中,尤其是多种免疫活性细胞中,均证实乙酰胆碱酯酶（acetylcholinesterase,AChE）、胆碱乙酰转移酶（choline acetyltransferase,ChAT）和乙酰胆碱受体（acetylcholine receptor,AChR）各亚型在内的胆碱能系统组分的存在,其中烟碱样乙酰胆碱受体α7（nicotinic acetylcholine receptor α7,nAChRα7）是烟碱样胆碱能抗炎通路（nicotinic anti-inflammatory pathway）中重要的分子核心机制,同时也是机体限制宿主防御反应扩大的内源性抗炎机制之一.本文旨在探讨（前）脂肪细胞上非神经元型胆碱能系统是否存在及初步揭示烟碱样胆碱能受体α7对前脂肪细胞功能的影响.以体外培养的3T3-L1前脂肪细胞为研究对象,采用免疫组化和蛋白质免疫印迹技术,分别检测前脂肪细胞和成熟脂肪细胞中乙酰胆碱酯酶、胆碱乙酰转移酶和烟碱样乙酰胆碱受体α7的3种胆碱能系统主要组分的蛋白表达.另将前脂肪细胞分为给予广谱烟碱样乙酰胆碱受体激动剂尼古丁、特异性烟碱样乙酰胆碱受体α7激动剂氯化胆碱及特异性烟碱样乙酰胆碱受体α7拮抗剂甲基牛扁亭碱干预12 h、24 h、36 h,并设立相应处理时间的空白对照组,逆转录聚合酶链反应检测前脂肪细胞visfatin mRNA表达情况.免疫组化染色可见前脂肪细胞中AChE、ChAT及AChRα7均有阳性表达;蛋白免疫印迹检测进一步半定量证实了前脂肪细胞和成熟脂肪细胞中AChE、ChAT及AChRα7的蛋白表达;拮抗剂甲基牛扁亭碱（10-6~10-4mol/L）时间、剂量依赖性上调前脂肪细胞visfatin mRNA表达（1.3~1.55-fold,P＜0.01）,与对应空白对照组相比,存在显著性统计学差异;加入不同剂量的尼古丁和氯化胆碱,则前脂肪细胞中visfatin mRNA表达水平与对应空白对照组相比,均不同程度地下降,其中以氯化胆碱的抑制效应更为显著.前脂肪细胞与成熟脂肪细胞中均存在有独立的胆碱能体系,其中AChRα7很可能在调节脂肪细胞因子分泌及肥胖相关的病理生理过程中发挥重要作用.

Objective To observe the expression of growth hormone secretagogue receptor-1a(GHSR-1a) gene during the period of 3T3-L1 preadipocyte differentiation and explore the relationship between GHSR-1a gene expression and adipocytes differentiation.Methods 3T3-L1 preadipocytes were cultured in vitro and differentiated into matured adipocytes.The morphological changes of 3T3-L1 adipocytes were observed and the differentiation rate was assayed by oil-red O staining,triglyceride(TG) mass was detected by chemical colorimetry method during the period of 3T3-L1 preadipocyte differentiation.Total RNA was extracted from adipocytes at varying time(day 0-8),and the level of GHSR-1a mRNA expression was measured by RT-PCR.Software of SPSS 12.0 was used to analyze all data.Results 3T3-L1 preadipocytes were fibroblastic and had no obvious fat droplet in cytoplasm.However,after they were induced to differentiate for 8 d,3T3-L1 preadipocytes became bigger and rounder,and differentiated into mature adipocytes with lots of fat droplets in the cells.Compared with that of preadipocytes,the concentration of TG mass began to increase at day 4 of differentiation and increased significantly at day 8 of differentiation(Pa0.01).GHSR-1a mRNA gene began to express in the preadipocytes and in the adipocytes of the first day of differentiation.From day 4-8 of differentiation,the GHSR-1a gene mRNA expression was obviously up-regulated.There were significant differences between any two detected time points in the levels of GHSR-1a gene mRNA expression(Pa0.05),except those points between day 0-1 and day 1-4(P0.05).Conclusions GHSR-1a gene mRNA expression is up-regulated during 3T3-L1 preadipocyte differentiation.It may participate in the adipocyte differentiation.Up-regulation in the level of GHSR-1a gene mRNA expression during 3T3-L1 adipocyte differentiation may be involved in the adipogenesis of adipocytes.

人生长激素(hGH)是脑神经垂体嗜酸性细胞分泌的一种蛋白质激素,它是体内最重要的促进生长的激素.儿童身高的增长主要是通过长骨骨干与骨骺之间的软骨板中的细胞分裂增殖实现的,生长激素(GH)正是对这种软骨细胞的分裂增殖具有显著的促进作用.