Objective:To compare the clinical characteristics, clinical efficacy and adverse drug reactions of rheumatoid factor (RF) positive (+ ) and negative (-) polyarticular juvenile idiopathic arthritis (PJIA).Methods:The clinical data of 67 PJIA patients admitted into Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, from January 2013 to December 2018 were analyzed retrospectively.They were divided into RF-positive PJIA group [RF (+ ) group, 23 cases] and RF-negative PJIA group [RF (-) group, 44 cases] according to RF titer.The clinical characteristics, laboratory indexes and clinical efficacy evaluation of the two groups were compared.Results:(1)Distribution of affected joints: the top 3 affected joints in the RF (+ ) group were the knuckles (16 cases, 69.57%), the wrists (15 cases, 65.22%) and the ankles (13 cases, 56.52%), and those in the RF (-) group were the knees (33 cases, 75.00%), ankle joints (29 cases, 65.91%) and hip joints (26 cases, 59.09%). The wrist joint involvement of the RF (+ ) group was significantly higher than that of the RF (-) group, while the knee joint involvement was lower than that of the RF (-) group.The difference was statistically significant (all P<0.01). (2)Magnetic resonance changes of the affected joints: articular cavity effusion (54 cases, 84.38%), synovial thickening (44 cases, 68.75%) and bone edema (26 cases, 40.63%) are common in both groups.The incidence of bone destruction (7 cases, 70.00%) and soft tissue edema (7 cases, 70.00%) in the RF (+ ) group was higher than that in the RF (-) group (2 cases, 18.18% and 2 cases, 18.18%), the difference was statistically significant (all P<0.05). (3) Changes in laboratory indicators: the positive rates of C-reactive protein, erythrocyte sedimentation rate, anti-cyclic citrullinated peptide antibody and anti-nuclear antibody in the RF(+ ) group were significantly higher than those in the RF(-) group, the difference was statistically significant (all P<0.05). (4)Juvenile arthritis disease activity score 27 (JADAS27): the score difference between RF(+ ) group and RF(-) group was not statistically significant [(22.83±5.60) scores vs.(23.07±6.66) scores, t=0.148, P>0.05]. (5) Efficacy analysis: 2 patients were lost to follow-up after discharge, and the remaining 65 patients were treated with traditional therapy, of which 30 were given biologics at the first hospitalization, 9 cases were treated with biologics after the failure of traditional treatments, and 35 patients were treated with biologics to control disease activity.In different dosage regimens, the disease remission rate in the RF(-) group is generally higher than that in the RF(+ ) group. Conclusions:PJIA patients have complicated joint involvement, RF-positive patients are more prone to joint destruction, and traditional treatments are less effective.Biological agents can effectively improve the symptoms of severe PJIA patients, especially those with poor prognosis.

Objective To evalute the affect on secretion levels and protein expression of the key enzyme of lipid metabolism(LPL, HSL, perilipin, DGAT), and the affect on protein expression of classical signaling pathways of insulin (IRS-1, PI3K) in 3T3-L1 adipocytes with acylation stimulating protein (ASP) stimulated under the condition which produce insulin resistance by free fatty acids. Methods The different concentrations of the Oleate and Palmitate(0, 0. 125, 0. 5, 1. 0mmol /L) were added to cultured 3T3-L1 adipocytes overnight. The real-time-PCR were used to detected mRNA of LPL, HSL, Perilipin, DGAT. The western blot were used to detected protein of HSL, prelipin, IRS-1, PI3K. Results After incubated with ASP, comparing with that of 0mmol /L, the levels of HSL, perilipin, LPL and DGAT mRNA expression were increased. In insulin and ASP stimulating condition, comparing with the 0mmol / L, the levels of HSL, perilipin, PI3K protein expression were increased. And comparing with the insulin stimulated, the protein expression of HSL, perilipin IRS-1 and PISK failed to significantly increase with the ASP stimulated. Conclusion Under the condition of insulin resistance, ASP can increase the mRNA expression of HSL, LPL, perilipin and DGAT by increasing the expression of these enzymes to improve the sensitivity and improve the use of the body's fat cell, to regulate the glucose and lipid metabolism. INS and ASP can increase the protein expression of the key enzyme of lipid metabolism (HSL, perilipin), and the classical insulin signaling pathway (PI3K).

目的 了解川崎病(KD)患病情况及临床特征,探讨KD冠状动脉损害(CAL)及IVIG耐药的危险因素.方法 回顾性分析华中科技大学同济医学院附属同济医院2012年1月1日至2016年12月31日初诊的KD患儿的临床资料,比较分析KD治疗前后,典型和不完全KD,KD伴或不伴CAL,IVIG敏感或耐药的临床特征,分析CAL发生和IVIG耐药的危险因素.结果 725例KD患儿进入本文分析,男:女为1.61:1,平均年龄(2.7±2.3)岁;不完全KD 206例(28.4％),典型KD 519例;CAL 216例(29.8％),IVIG耐药61例(8.4％);治疗中仅使用阿司匹林者70例(9.6％).KD伴CAL的危险因素为IVIG耐药(OR=5.138,95％CI:1.835～14.836)和氨基末端脑钠肽前体(NT-proBNP)≥1000 pg·mL-1(OR=2.723,95％CI:1.110～6.679).IVIG耐药的危险因素为出现CAL(OR=2.586,95％CI:1.067～6.271).结论 KD患病人数、CAL和IVIG耐药患儿有增加趋势.IVIG耐药和NT-proBNP≥1000 pg·mL-1为KD伴CAL的危险因素,而发生CAL为IVIG耐药的危险因素.

Objective To detect serum interleukin 6(IL-6), N-terminal pro-brain natriuretic peptide (NT-proBNP) and serum ferritin in acute Kawasaki disease (KD), and explore their values in the diagnosis of KD, and further to explore the relationship with intravenous immunoglobulin (IVIG) unresponsiveness and coronary arterial lesions (CALs). Methods Totally 108 patients with KD (81 IVIG responders and 27 IVIG non-responders, 31with CALs and 77 non-CALs) were recruited from October 2014 to February 2016 at Department of Pediatrics of Tongji Hospital affiliated to Tongji Medical College, Huazhong University of Science and Technology, 64 were boys and 44 were girls. Their ages ranged from 2 months to 11 years and 5 months. A total of 30 children with respiratory tract infection were selected as the control group, 18 were boys and 12 were girls, ages ranged from 4 months to 10 years. Serum IL-6, NT-proBNP and serum ferritin were measured at the day of admission. The differences between groups were analyzed by t-test. To compare the power of serum level of interleukin 6(IL-6), N-terminal pro-brain natriuretic peptide (NT-proBNP) and serum ferritin levels in predicting KD, IVIG unresponsiveness and CALs, receiver-operating characteristic (ROC) curves were plotted and areas under the curve (AUC) were calculated. All data are presented as means ± standard deviation. Results (1) The levels of IL-6 135 ± 268ng /L, NT-proBNP 1008 ± 1675ng /L and ferritin 227 ± 238μg /L were significantly higher in the acute phase patients with KD than those of the control group 27 ± 29ng /L for IL-6 (t = 2. 192, P = 0. 03), 109 ± 100ng /L for NT-proBNP(t = 5. 463, P = 0. 000) and 72 ± 101μg /L for ferritin (t = 3. 437, P = 0. 001). (2) The levels of NT-proBNP 1837 ± 2666ng /L in IVIG unresponsive group were significantly higher than those of the IVIG responsive group 720 ± 1032ng /L (t = 3. 108, P = 0. 002). However, there were no significant difference of IL-6 and serum ferritin between the two groups. (3) The levels of NT-proBNP in CALs group 1703 ± 2569ng /L vs 742 ± 1080ng /L, serum ferritin 340 ± 405μg /L vs 183 ± 99μg /L were significantly higher than those of the non-CALs group (P ＜ 0. 05). However, there was no significant difference of IL-6 between the two groups. (4) The area under the curve for predicting KD with various variables were as follows: serum IL-6 0. 773, NT-proBNP 0. 835 and serum ferritin 0. 793. The area under the curve for predicting resistance to IVIG with serum NT-proBNP was 0. 623. The area under the curve for predicting CALs with various variables were as follows: NT-proBNP 0. 612 and ferritin 0. 671. The ROC of ferritin for predicting CALs is better than NT-proBNP. A ferritin cut-off value of 160. 2μg /L yielded a sensitivity of 73. 7％, specificity of 52. 1％. Conclusion The serum IL-6, NT-proBNP and serum ferritin can be used as useful parameters in early diagnosis of KD. Elevated NT-proBNP or serum ferritin may be useful to predict IVIG resistance and CALs in KD patients.

Kawasaki disease(KD) is an acute,self-limited vasculitis of childhood and has become the leading cause of acquired pediatric heart disease.The underlying etiology remains unknown.The disease itself may be the characteristic manifestation of a common pathway of immune-mediated vascular inflammation in susceptible hosts.Intravenous immunoglobulin (IVIG) is now widely accepted as the first-line therapy for KD.However,approximately 10％-20％ of patients are resistant to IVIG.Although,an additional administration of IVIG is often chosen for the treatment of KD patients resistant to the first administration of IVIG,its efficacy is reported to be lower than that of the first IVIG dose.Thus,it is clear that some KD patients can not be treated successfully by IVIG alone,even if it is used repetitively.Currently,methylprednisolone(MP) is used for the treatment of IVIG-resistant KD.However,the proper use of MP for maximum effect and safety has not yet been elucidated.Tumor necrosis factor-α blocker,infliximab effective in the control of inflammation in patients with resistant KD.Cyclosporin treatment is also a promising option for patients with refractory KD.Plasma exchange was a safe,effective prophylactic measure against coronary artery lesions in children with KD refractory to intravenous gamma globulin therapy.

In addition to skin and muscle lesions,severe juvenile dermatomyositis can also cause multiple systemic and organ damage,which directly threatens the life of children.The development of imaging and specific antibody detection techniques is beneficial to the early diagnosis of severe juvenile dermatomyositis.Early diagnosis and regular combination of continuous medication can improve its prognosis.At present,the treatment of severe juvenile dermatomyositis is mainly based on large doses of glucocorticoid.Intravenous immunoglobulin is a rapid and effective treatment and can be used as a first-line drug.It can be repeated in half a year.For children with combined organ damage,selective use of cyclosporine,mycophenolate mofetil,tacrolimus and other immunosuppressive agents,even biological agents,plasma exchange,stem cell transplantation,etc.,but the efficacy needs to be further verified.

目的 观察3T3-L1成熟脂肪细胞胰岛素抵抗状态下促酰化蛋白(acylation stimulating protein,ASP)对炎性因子(IL-6、MCP-1、MIP-1α和TNF-α)分泌水平以及炎性信号因子(JNK1、IKKβ)蛋白表达的影响.方法 3T3-L1成熟脂肪细胞分别给予不同浓度(0、0.125、0.5、1.0mmol/L)油酸(C18:1)或棕榈酸(C16:0)温育过夜,诱导胰岛素抵抗,在此基础上给予ASP刺激,用ELISA测定IL-6、MCP-1、MIP-1α和TNF-α 4种细胞炎性因子的分泌水平,采用Western blot法检测JNK1和KKβ蛋白表达.结果 油酸诱导的胰岛素抵抗下,ASP刺激的脂肪细胞IL-6和MCP-1的分泌水平轻度下调,但差异无统计学意义;而ASP刺激的脂肪细胞MIP-1α和TNF-α的分泌水平显著下降,MIP-1α和TNF-α的分泌分别下调57％(P＜0.05)和48％ (P ＜0.05)(1.0mmol/L油酸组).棕榈酸诱导的胰岛素抵抗下,ASP刺激的脂肪细胞IL-6分泌水平呈下降趋势,最大下调50％ IL-6(P＜0.05);22％ MCP-1(P＞0.05),35％ MIP-1α(P ＞0.05)和38％ TNF-α(P ＞0.05)的分泌.高浓度(1.0mmol/L)油酸和棕榈酸诱导的脂肪细胞胰岛素抵抗状态下,ASP刺激的炎性信号蛋白JNK1蛋白表达显著下调,分别为34％ (P ＜0.05)和54％ (P ＜0.01).但IKKβ蛋白表达差异无统计学意义.结论 在脂肪酸诱导的脂肪细胞胰岛素抵抗状态下,ASP在一定程度上调节炎性因子分泌,参与调节JNK、IKK/NF-κB信号通路中重要信号分子的功能,ASP参与了脂肪细胞脂毒性-炎性反应的调控.

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.

Aim To examine the effect of estradiol,testosterone and progesterone on Visfatin gene and protein expression in cultured 3T3-L1 adipocytes and preadipocytes.Methods 0 mmol/L(sex hormones-free DMEM/F12),10-8 mol/L,10-7 mol/L and 10-6 mol/L estradiol or testosterone or progesterone were added to cultured 3T3-L1 adipocytes or preadipocytes overnight.Total RNA and proteins were extracted.Then the expression of Visfatin mRNA and protein was measured by RT-PCR and Western blot,respectively.Results Overnight incubation with estradiol increased Visfatin mRNA expression in both adipocytes(24% to 21%,P<0.05) at the concentration of 10-8 mol/L to 10-7 mol/L and preadipocytes(70% to 123%,P<0.01) at 10-7 mol/L to 10-6 mol/L,respectively.Visfatin mRNA expression increased by 28%(P<0.05) at 10-8 mol/L testosterone in adipocytes,76%(P<0.05) at 10-7 mol/L testosterone in preadipocytes.Progesterone did not affect Visfatin mRNA expression on adipocytes,but had a marked increase in Visfatin expression in preadipocytes by 2.6-fold(P<0.05).In 3T3-L1 adipocytes,estradiol had small but insignificant effects on Visfatin protein expression,whereas testosterone had a much greater effect,increasing Visfatin protein by 134% at 10-6 mol/L(P<0.05).However,progesterone significantly decreased Visfatin protein by 32% at 10-6 mol/L(P<0.05).Conclusions These data suggest that Visfatin may play a physiological role in the insulin resistance caused by estradiol,testosterone or progesterone in 3T3-L1 adipocytes and preadipocytes.

OBJECTIVE To explore the role of adipokines including insulin, resistin, leptin, adiponectin, acylation stimulating protein (ASP) and complement C3 (C3) in various types of obesity (peripheral obesity, abdominal obesity and mixed obesity) in Chinese children and adolescents, and their relationships with body size and pubertal development. METHODS Children and adolescents (n=3 508) aged 6 to 18 years, with 1 788 boys and 1 720 girls were assessed for body mass index, waist circumference, pubertal development, blood insulin, resistin, leptin, adiponectin, ASP and C3 levels. Three types of obesity [peripheral obesity (n=43), abdominal obesity (n=473), mixed obesity (n=1 187)] and non-obese control (n=1 805) were defined with combined use of Chinese body mass index and waist circumference criteria. RESULTS Serum resistin, leptin and adiponectin levels were higher in girls than those in boys (all P<0.01). Insulin and leptin increased and adiponectin decreased across five Tanner stages in both girls and boys (all P<0.001), while ASP changed only in girls (P<0.001) and C3 only in boys (P<0.001). Insulin, leptin and ASP were higher, but adiponectin was lower in all three types of obesity vs. the non-obese control (all P<0.05). The greatest abnormalities of all six adipokines were found in the mixed obesity group. With inclusion of body mass index and waist circumference in simultaneous regression analyses, both body size indices were independently and significantly correlated with insulin, leptin and adiponectin after age and gender adjustment. Compared with waist circumference, the body mass index was stronger in interpreting insulin, leptin, adiponectin and ASP levels, whereas it was weaker in explaining variance of plasma C3. CONCLUSION Obese children have a worse metabolic profile with high insulin, resistin, leptin, ASP and C3, and low adiponectin levels. The adipokine profile in mixed obesity is worse than that in peripheral or abdominal obesity. Identification of obese subjects with a malignant adipokine profile using a combination of body mass index and waist circumference is important for the prevention of obesity-related disease.

小儿肥厚型心肌病(HCM)为一种单基因疾病,多数呈常染色体显性遗传,主要基因编码在肌节蛋白;小儿HCM的病因和预后与成人不尽相同,除特发性外,可见先天代谢缺陷、各种畸形综合征、神经肌肉疾病等引起的HCM.现阐述小儿HCM的病理和病理生理、临床表现及诊断方法,并介绍药物治疗的进展和Morrow 心肌切除术、经皮室间隔消融术、双腔起搏以及植入式心脏复律除颤器的适应证.

胰岛素抵抗不仅是2型糖尿病的发病基础,更是贯穿多种代谢相关性疾病的主线。糖、脂代谢是生物最基本的能量代谢形式。近年来研究发现,许多胰岛素抵抗都伴随着某些炎性因子以及游离脂肪酸水平的升高,胰岛素抵抗不是机体糖脂代谢稳态失衡的唯一表现形式,胰岛素抵抗的概念不仅仅是狭义的胰岛素自身,而应是由多种脂源性激素参与的"胰岛素抵抗状态",这种状态实质上代表一种普遍意义的脂肪抵抗状态。因此,"脂毒性-炎症反应"与脂源性激素抵抗的关系正成为当前研究的热点,本文就二者关系作一综述。

OBJECTIVE The current standard method of treatment of Kawasaki disease is the use of high doses of gamma globulin(2 g·kg-1) and aspirin.This study will use the Meta analysis of the evidence-based medicine to explore the advantages and disadvantages of either glucocorticosteroid or repeated intravenous gamma globulin in treating Kawasaki disease after failure of initial treatment.METHODS This study selected six research groups which were related to the patients with Kawasaki disease who failed to respond to the initial intravenous gamma globulin treatment;then used the Meta analysis to compare the two kinds of treatment in the aspects of damage of the coronary arteries,fever duration and serum CRP levels.RESULTS 6 research groups included 156 patients for analysis.89 patients were received intravenous glucocorticosteroid and 67 cases were given repeated intravenous gamma globulin.In comparison of intravenous glucocorticosteroid versus repeated intravenous gamma globulin,OR for the damage of the coronary arteries was 0.65(95%CI,0.29-1.45)0 with rhombus intersection with the vertical line,WMD for fever duration was-1.91(95%CI,-2.88——0.95)0 with rhombus situated to the left side of the vertical line.WMD for the serum CRP levels was-3.94(95%CI,-5.42——2.45)0 with rhombus entirely situated to the left side of the vertical line.Therefore,both efficacy in reducing coronary artery injury were no significant difference,but in terms of fever and reduce the effect of CRP were the significant differences.CONCLUSION Glucocorticoid can play a better therapeutic effect and spend less in the economy than re-use of gamma globulin in the failure of initial treatment of Kawasaki disease.

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 evaluate potential acylation stimulating protein(ASP) resistance in both adipocytes and preadipocytes under the conditions of insulin resistance induced by testosterone on both receptor level and post-receptor level.Methods: 3T3-L1 preadipocytes were cultured and differentiated,and the adipocytes and preadipocytes were treated with 0(testosterone-free DMEM/F12),10-8,10-7,and 10-6 mol/L testosterone overnight.RT-PCR and flow cytometry were used to detected mRNA and cell surface expression of ASP receptor.Both non-testosterone treated and testosterone treated 3T3-L1 cells were cultured with 5.0 μmol/L ASP for 4 hours.Then the cell proteins were extracted and the expressions of Gβ,Gαq/11,p-PKCα,and p-PKC ζ were measured by Western blot.Results: High dose testosterone suppressed the C5L2 mRNA and protein expression in 3T3-L1 adipocytes but not in preadipocytes.At 10-6 mol/L,testosterone inhibited C5L2 mRNA and cell surface C5L2 expression by 60%(P<0.01) and 27%(P<0.01),respectively.After overnight incubation with testosterone(in adipocytes and preadipocytes),Gαq/11,Gβ,p-PKCα,and p-PKCζ were downregulated in the presence of ASP treatment to a certain degree.In adipocytes,testosterone effectively blocked the ASP-stimulated Gαq/11,p-PKCα,and p-PKCζ expression by 52%,50% and 57%(P<0.05 to P<0.01,respectively) at 10-6 mol/L in adipocytes.In preadipocytes,testosterone did not influence the four proteins' expression.Conclusion: Testosterone inhibited ASP-stimulated signal proteins.The ASP resistance mechanism of action involves both changes in expression of C5L2 as well as signaling parameters to some extent.Testosterone-induced ASP resistance may contribute to the physiological abnormalities associated with insulin resistance and some abnormalities related with testosterone such as disorders in polycystic ovarian syndrome and so on.

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.

Objective To investigate the relationship between plasma acylation stimulating protein(ASP) and obesity and metabolic syndrome(MS) among Chinese children and adolescents.Methods Based on the Beijing Child and Adolescent Metabolic Syndrome(BCAMS) study,1 603 children and adolescents(873 boys and 730 girls) aged from 6 to 18 years old were randomly selected from the control group(without any MS component) and the case group(with at least one of the five MS components),and divided into three groups of normal weight(n=603),overweight(n=291) and obese(n=709) according to the sex-age-specific body mass index cutoffs recommended by Chinese Working Group on Obesity(WGOC).The presence of pediatric MS was defined with modified NCEP ATPⅢ criteria by the presence of three or more items of the following five components:①central obesity was defined as waist circumference≥90th percentile for age and gender(established according to the BCAMS study);②elevated systolic and/or diastolic blood pressure ≥90th percentile for age and sex(according to the BCAMS study);③hypertriglyceridemia was defined as TG≥1.24 mmol·L-1;④low serum high-density lipoprotein cholesterol(HDL-C) was defined as ≤1.03 mmol·L-1;⑤impaired fasting glucose(IFG) was defined as 5.60 mmol·L-1.Data including anthropometric measurements(height,weight and waist circumference),systolic and diastolic blood pressure,fat mass percentage by bioimpedance analysis,pubertal development stage were collected.Venous blood samples were obtained by direct vein puncture after an overnight(minimum 12 h) fast.Plasma glucose was detected by the glucose oxidase method.Serum total cholesterol,triglyceride,HDL-C and low-density lipoprotein cholesterol(LDL-C) were measured.Plasma ASP was measured by enzyme-linked immunosorbent assay(ELISA),and plasma complement 3(C3) concentration was determined by turbidimetric assay using a polyclonal anti-human antibody specific against C3(Lin-Fei Co,P.R.China).For C3 and ASP,the intra-and inter-assay coefficients of variations were 4% and 8%.The percentage of C3 converted to ASP(%ASP/C3) was calculated to evaluate the extent of conversion C3 to ASP.One-way analysis of variance(ANOVA) and general linear model were used to compare the levels of plasma ASP and C3 among various weight status groups.The odds ratios(ORs) and 95% confidence intervals(CIs) were analyzed with multivariable Logistic regression model to investigate the effect of ASP on metabolic abnormalities.Results Within the total groups,376 subjects were with the presence of MS,and the prevalence rates were 2.2%,15.5% and 44.9% in normal weight,overweight and obese groups,respectively.Girls had higher plasma ASP than boys(t=2.527,P0.05).Plasma ASP levels were increased in overweight group and obese group compared with in normal weight group in boys(geometric mean:57.88 nmol·L-1,60.63 nmol·L-1 vs 37.58 nmol·L-1,F=34.329,P0.001) and girls(geometric mean:60.25 nmol·L-1,66.68 nmol·L-1 vs 44.16 nmol·L-1,F=22.246,P0.001).C3 was higher in obese group than that in normal weight group only in girls(P0.01).The percentage of C3 converted to ASP(%ASP/C3) was also increased in overweight group and obese group versus in normal weight group in both boys(F=17.382,P0.001) and girls(F=6.317,P=0.002).Plasma ASP increased with the clustering of MS components in boys(F=16.422,P0.001) and girls(F=9.661,P0.001).Increased ASP was also associated with the presence of one single metabolic abnormality(especially hypertension,abdominal obesity or hyperglycemia).With age,gender and pubertal development as covariants and the lowest quintile of plasma ASP as reference,the ORs(95%CIs) of the highest quintile of ASP were 3.90(2.38-6.39),6.05(4.06-9.01) and 2.89(1.93-4.33) for predicting overweight,obesity and MS,respectively.But the statistical significance of OR(95%CI) for MS was vanished(P0.05) after further adjustment for body mass index.Conclusions Elevated plasma ASP levels in obese children and adolescents are associated with the clustering of MS components,and distinct metabolic abnormalities.The level of plasma ASP may be a predictor of overweight,obesity and MS in children and adolescents.

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.

OBJECTIVE:Childhood obesity is increasing worldwide and is increasingly associated with metabolic syndrome (MetS). Our aim was to examine acylation stimulating protein (ASP) and its precursor complement C3, in normal, overweight, and obese Chinese children and adolescents, and the relationships with body size, blood parameters, pubertal development, family environment, and MetS.METHODS:Children and adolescents (n=1603) from 6 to 18 years, boys (n=873) and girls (n=730), including normal weight (n=603), overweight (n=291) and obese (n=709) were assessed for body size parameters, pubertal development, blood lipids, glucose, insulin, ASP, and C3.RESULTS:ASP levels were increased in overweight and obese versus normal weight (P<0.001), while C3 showed little variation. This effect of overweight/obesity remained throughout early stages when boys and girls were separated by pubertal development or age, although age and pubertal status itself had no effect. Separation based on ASP quintiles demonstrated significant associations with blood cholesterol, triglyceride, low-density lipoprotein cholesterol (LDL-Chol), glucose, insulin, and homeostatic model assessment of insulin resistance in boys, and LDL-Chol, high-density lipoprotein cholesterol, and glucose in girls. A positive correlation with mother's body mass index in boys and girls (P=0.002 and P=0.014 respectively) as well as birth weight (P<0.001) was noted. MetS was strongly associated with increased ASP, the presence of a single MetS factor (especially hypertension, central obesity, or hyperglycemia) was associated with increased ASP.CONCLUSION:Changes in the plasma adipokine ASP in early obesity are associated with blood lipid and glucose modifications, family environment, and distinct MetS risk factors.

Acylation stimulating protein (ASP) stimulates triglyceride synthesis and glucose transport via its receptor C5L2. In human studies, ASP is increased in insulin resistant states such as obesity, diabetes, polycystic ovary syndrome and late pregnancy (the latter two associated with altered sex hormones). The aims were (i) to evaluate ASP response and C5L2 expression following treatment with sex steroid hormones and (ii) to identify mechanisms of ASP resistance using 3T3‐L1 adipocytes and preadipocytes. Overnight incubation with physiological progesterone (PROG) concentrations induced dose‐dependent inhibition of ASP‐stimulated glucose transport in adipocytes (188 ± 11% +ASP, 100 ± 4% control, 129 ± 18% to 85 ± 7% [ASP + PROG 10−8 to 10−6 M] and preadipocytes (263 ± 18% +ASP, 100 ± 3% control, 170 ± 11% to 167 ± 4% [ASP + PROG 10−8 to 10−6 M]), while estradiol and testosterone (TEST) were effective only at the highest concentration (10−6 M). In adipocytes, dose‐dependent maximal C5L2 mRNA decreases were 39–75% (P = 0.003), with decreased cell‐surface C5L2 of −22% and −27% (10−6 M PROG and TEST, respectively) with no change in preadipocytes. Adipocytes treated with PROG displayed decreases in G proteins: Gβ (−55%), Gαq/11 (−56%) as well as complete inhibition of ASP stimulation. PROG significantly decreased basal levels of phosphorylated PKCα (p‐PKCα) while there was no change in p‐ PKCζ. ASP increased p‐PKCα and PKCζ to 161% (P < 0.0.001) and 160% (P < 0.01), a stimulation effectively blocked by PROG (10−8 and 10−6 M) and TEST (10−6 M). Sex steroid hormone‐induced ASP resistance via C5L2 may contribute to altered adipose tissue function and insulin resistance phenotype in humans. J. Cell. Biochem. 105: 404–413, 2008. © 2008 Wiley‐Liss, Inc.