Objective To investigate the effect of RBP on 3T3-L1 adipocyte differentiation,lipid metabolism and glucose transporter 4(GLUT-4)gene expression.Methods We constructed an expression vector for rat resistin gene and transfected it into 3T3-L1 adipocytes.RBP was added to the medium of 3T3-L1 adipocytes or resistin-overexpressing adipocytes on day 0 of differentiation.Cell differentiation and lipid accumulation were determined by oil red O staining.The mRNA expressions of differentiation marker genes(pref-1,C/EBP?,FAS)and GLUT-4 gene were evaluated by RT-PCR.Triglyceride(TG)and free fatty acids(FFAs)in adipocytes were measured by colorimetric kit.Results(1)When 10-12mol/L RBP was applied,the percent of living cells was high and the shape was unchanged.(2)RBP had no effect on the differentiation of normal adipocytes,but significantly decreased the number of lipid droplets in resistin-overexpressing adipocytes without affecting the lipid droplets-presenting day.(3)C/EBP? and FAS expressions in resistin-overexpressing adipocytes were down-regulated after RBP was applied,without changing their expressions in normal adipocytes.(4)RBP had no effect on the cellular TG and FFAs levels in normal cells,whereas it can significantly decrease the levels in resistin-overexpressing adipocytes.(5)There was no difference in the expression of GLUT-4 gene between 3T3-L1 adipocytes and RBP-applied cells.Conclusions(1)RBP has no effect on the cell differentiation and lipid metabolism in normal 3T3-L1 adipocytes.(2)RBP can inhibit the cell differentiation and lipid metabolism of resistin-overexpressing 3T3-L1 cells.(3)RBP has no effect on the expression of GLUT-4 gene.

BACKGROUND:Obesity is an important problem concerned domestically and internationally. How to control the body mass and to reduce the weight without any effect on normal food intake is the focus for study.OBJECTIVE:To probe into the weight-reducing effect of Oolong tea and its effect on the mRNA level of β3-adrenergic receptor(β3-AR).DESIGN: Completely randomized grouping, controlled trial SETTING:Department of Nutrition and Food Hygiene, College of Public Health, Nanjing Medical University; Institute of Pediatric Medicine, Nanjing Medical University.MATERIALS: This experiment was conducted in the laboratory of Nanjing Medical University from September 2003 to February 2004. The obese rat models were made with the diet of high energy and high fat in male rats weighing about 80 g. Thirty-two male obese rats were selected.And Oolong tea extract was prepared, whose concentration was equivalent to 0.24 g of tea.INTERVENTIONS:Thirty-two male obese rats were divided randomly in-to 4 groups: obese control group, low, middle and high dose of oolong tea groups. There were 8 rats in each group. The rats in the control group were fed by gavage with distilled water every day. The other rats in low, middle or high dose of Oolong tea groups were fed by gavage with 0.4 g/kg, 1.2 g/kg, and 2.4 g/kg of Oolong tea respectively. They were all fed with diet of high energy and high fat. Each rat was raised in separate cage. The room temperature for the rats remained about 22 ℃ with the humidity of 55%. The rats were free access to water, but the diet was fed twice a day at a fixed amount. If it was finished, no more diet would be added. Thirty days later, body mass, maximal diameter of adipocytes were measured, and β3-ARmRNA levels in adipose tissues were measured.MAIN OUTCOME MEASURES: Body mass, increased body mass,the weight of adipose tissues in retroperitoneal, peri-epididymal and interscapular regions and the maximal diameter of adipocytes were measured,β3- AR mRNA levels in the adipose tissues above were measured with the method of RT-PCR.RESULTS:According to intention-to-treat analysis, thirty-two male rats entered result analysis. [1]Body mass: Increased weight of rats in 1.2 g/kg and 2.4 g/kg Oolong tea groups were significantly lower than that in the rats of control group and rats in 0.4 g/kg Oolong tea group (58.4±46.7,68.1±30.4,125.7±34.4,96.3±26.2,P ＜ 0.01), but the amount of total diet consumption was similar in each group (P ＞ 0.05). [2]Lipid coefficient in retroperitoneal and peri-epididymal regions of rats in 1.2 g/kg and 2.4 g/kg Oolong tea groups was lower than that in the rats of control group and rats in 0.4 g/kg Oolong tea group,(1.57±0.53,2.14±0.90 to 2.71±0.49,2.50±0.53, 1.14±0.38,1.43±0.53 to2.00±0.00,1.88±0.35), but there was no significant difference among groups of the ratio in inter-scapular regions (P ＞0.05). [3]The maximal diameter of adipocytes: The maximal diameter in retroperitoneal, periepididymal and inter-scapular regions of the rats in 0.4 g/kg, 1.2 g/kg and 2.4 g/kg Oolong tea groups was significantly lower than that in the rats of control group[(113±24), (86±29), (90±23), (120±30)μm;(94±20), (80±18), (64±17), (111±21)μm; (24±11), (21 ±11), (22±10),(27±11)μm,P ＜ 0.05]. [4]β3-AR mRNA levels in adipose tissues:The β3-AR mRNA levels in retroperitoneal, peri-epididymal and interscapular regions of the rats in 1.2 g/kg and 2.4 g/kg Oolong tea groups were significantly higher than those in rats of control group and rats with0.4 g/kg of Oolong tea (0.72±0.11,0.64±0.112,0.40±0.08,0.34±0.10 for retroperitoneal region, 1.06±0.21,1.02±0.24,0.42±0.15,0.43±0.11 for epididymal region, 1.01±0.42,0.70±0.17,0.42±0.10,0.49±0.16 for interscapular region, P ＜ 0.01).CONCLUSION:Oolong tea was of weight～reducing effect,which may be related to its effect to increase β3-AR mRNA level The middle dose (1.2 g/kg) may be optimal.These results may be helpful to the theory of weight reducing with tea and the selection ofoptimal dose.

ObjectiveTo investigate the effects of erythro po ietin on prevention hypoxic-ischemic brain damage (HIBD) and activation of Casp ase-3 in Hippocampal CA1 region in newborn rats. Methods7 d Sprague-Dawley rats were divided into hypoxic-ischemic (HI) group (n=11), recombinant human erythropoietin (rhEPO) treated group (n=11), and sham-op erated control group (n=9). HIBD was established in both HI group and rhEPO treated group. The number of rats animals with spontaneous left-turn in two gro ups was counted respectively at subsequent different time: 0, 6, 12 and 24 h. Th e expression and distribution of activated Caspase-3 was detected by immunohist ochemistry analysis. The positive cells were calculated in hippocampal CA1 regio n of every groups.ResultsTwo rats in HI group and one in rhE PO treated group died from continuous convulsion during hypoxia. all survival ra ts in up two groups had spontaneously left-turn Compared with HI group, the r ate of spontaneous left-turn was dramatically lower in rhEPO treated group (HI group vs rhEPO treated group, 1/10 vs 6/9, P=0.0198) at 24 h after hypox ia. The positive stained cells were distributed dispersively in the brain of con trol group, and more intensively in the hippocampus and cerebral cortex of the o ther two groups. In CA1 region, the number of positive cells in HI group, was si gnificant higher than that in control group ( 41.38 ?2.09 vs 10.52?2.70 , P

Objective To explore the status of C57BL/6J mouse brown fat adipogenic differentiation function with aging.Methods C57BL/6J female and male mice at the ages of 0-week (newborn),4-week,8-week,12-week old were selected from the same brood,brown adipose tissue was obstained from their interscapular region,and the brown adipose was identified by using immunohistochemical markers.Then the total RNA was extracted from the brown adipose and quality identification was determined at the same time.The expression levels of the related genes (PPARα,C/EBPα,PGC-1α,PPARγ,FOXC2,BMP7) induced by brown adipose adipogenic differentiation were detected by quantitative real-time PCR in 0-week,4-week,8-week,12-week mice.Results Uncoupling protein -1 (UCP1) immunohistochemical data indicated that positive deep-colour substance was brown adipose tissue.Quantitative Real-time PCR also indicated that the expression volume of adipogenesis gene gradually reduced with aging,and there were significant differences at the different time points [PPARα (F =11.96,P ＜ 0.000 1),C/EBPα (F =9.39,P ＜0.000 1),PGC-1α(F =17.21,P ＜0.000 1),PPARγ(F =13.11,P ＜0.000 1),FOXC2(F =12.23,P ＜0.000 1),BMP7(F =16.44,P ＜0.000 1)].Conclusions The adipogenic differentiation ability and activity of mouse brown adipose gradually reduce with aging.But the regulatory factors for its function needs to be further investigated.

Objective To establish an effective DNA isolation method for neonatal disease screening,so as to explore its application to the methylation detection.Methods The 20 dried blood spots samples were randomly divided into 2 groups according to the gender:the traditional method group (n =10) and the improved kit method group(n =10).The DNA quality was evaluated based on its concentration,integrity and whether it could be used in polymerase chain reaction (PCR).These DNA samples with or without bisulfite treatment were used as template in the methylation-specific polymerase chain reaction (MSP).The methylation levels of Leptin and tumor necrosis factor-α (TNF-α) gene promoter region were detected.Results DNA concentration of the improved kit method [(5.70 ± 0.81) mg/L] was significantly higher than that of the traditional method [(3.50 ± 0.45) mg/L] (t =2.79,P ＜ 0.05),and biochemical analyzer analysis showed a better DNA integrity.Agarose gel electrophoresis revealed that 18S gene fragment could be successfully amplified by PCR method,suggesting its potential application to PCR study.MSP results showed different DNA methylation levels of Leptin and TNF-α genes promoter regions from various samples.Conclusions The improved kit method can effectively extract DNA from dried blood spots samples,and these DNA can be used in methylation research.The study can provide a new research direction and technical method to reveal the pathogenesis of disease from the perspective of DNA methylation.

Objective To explore the influence of middle cerebral artery blood flow on mechanical ventilation in very low birth weight premature after using Budesonide(BUD) mixed with pulmonary surfactant(PS),and to explore the protection mechanism of cerebral injury.Methods Forty premature infants (gestational age ＜ 34 weeks,birth weight ＜ 1 500 g) with respiratory distress syndrome(RDS) were randomly assigned into study group and control group in Nanjing Maternal and Child Health Hospital from Aug.2010 to Mar.2012.PS and BUD mixture was used in study group (Per 70 mg PS adding BUD 0.25 mg),PS dose of 70 mg/kg,BUD dose of 0.25 mg/kg.Control group was only administered with PS,dose 70 mg/kg.It was administered intratracheally after 30 to 60 minutes of birth in both groups.The index of blood flow rate and blood vessel elasticity of arteria cerebri media [including systolic velocity (Vs),diastolic velocity (Vd),mean velocity (Vm),resistant index (RI) and elasticity index (PI)] were monitored by using transcranial Doppler.Results The Vs increased steadily in study group,but instability in control group,and there were of statistical differences on the 4 d,5 d,6 d and 7 d (t =3.21,2.95,3.12,3.43,all P ＜ 0.05).The Vd increased steadily in study group,but unsteadily in control group,and there were statistical differences on the 4 d,5 d,6 d and 7 d (t =4.21,3.10,3.98,4.56,all P ＜0.05).The Vm of study group was higher than that in the PS group,and there were statistical differences on the 4 d,5 d,6 d and 7 d (t =2.68,2.98,3.98,3.57,all P ＜ 0.05).The RI of study group was higher than that in the control group,and there were statistical differences in the 5 d,6 d and 7 d(t =3.10,3.98,4.06,all P ＜ 0.05).PI steadily in study group,but instability in control group,and there were statistical differences in the 5 d,6 d and 7 d (t =4.18,3.23,3.02,all P ＜ 0.05).The overall incidence of periventricular/intraventricular hemorrhage showed no significant difference,but severe periventricular/intraventricular hemorrhage (grade Ⅲ,Ⅳ) of study group was less than that in the control group (x2 =4.80,P ＜ 0.05).The incidence of periventricular leukomalacia was reduced in the study group compared with that in the control group (x2 =3.31,P ＜ 0.05).Conclusion The very low birth weight infants treated with mechanical ventilation show steady cerebral blood flow and lower incidence of brain injury after using BUD mixed with pulmonary surfactant.

Objective To explore the mechanism of intrauterine growth restriction (IUGR) via observing the change of mitochondria in IUGR placenta. Methods Placenta samples were collected from 30 singleton pregnancies at the time of elec-tive caesarean section. Fifteen of them were appropriate for gestational age and 15 were IUGR. Mitochondrial morphology was observed by transmission electron microscopy, DNA copies were analyzed by real-time quantitative PCR and membrane potential was assayed by lfow cytometry. Results Signiifcant morphological changes of placental mitochondria were observed under transmission electron microscopy in IUGR, mitochondrial DNA copies in IUGR placenta were signiifcantly increased (P<0.01) and membrane potential decreased dramatically (P<0.01). Conclusions It is suggest that impaired mitochondrial function in IUGR may involve in IUGR pathogenesis.

Objective To predict the functions of hsa-miR-1908 promoter using various bioinformatic tools, and to provide clues for further study on transcriptional regulation mechanism of miR-1908 in human adipocytes. Methods The promoter se-quence of miR-1908 was obtained from Ensemble, and then the CpG islands and transcription factor binding sites were pre-dicted by a variety of online bioinformatic tools. Results The length of the miR-1908 promoter sequence was 1 458 bp. The CpG islands, which inhibited the transcription of miR-1908, were located at (438-756) bp, (836-937) bp and (979-1374) bp. Meanwhile, 15 transcription factor binding sites were found in the promoter sequence of miR-1908. Conclusions miRNA up-stream promoter related bioinformatics can not only improve the efficiency of microRNA promoter research, but also provide further important information on transcriptional regulation of miR-1908.

Objective To observe the effect of Triclosan( TCS) exposure on Caenorhabditis elegans( c. ele-gans) F1 generation of locomotory behavior, brood size, and generation time. Methods The trial included a control group and 4 TCS treatment groups with different doses (100 nmol/L,1 μmol/L,10μmol/L,20μmol/L),the exposure time being 24 hours,the effect of c. elegans′head thrashes,body bending frequency,the brood size and generation time was observed. Results (1) The control group exposed to 100 nmol/L,1 μmol/L,10 μmol/L,20 μmol/L TCS,their head thrash frequency of c. elegans F1 was(109. 40±8. 61) times/min,(84. 70±7. 82) times/min,(76. 35±7. 44) times/min,(74. 74±5. 93)times/min,(71. 95±4. 19)times/min,respectively,the head thrash ability of c. elegans was significantly inhibited(F=62. 245,P<0. 01). (2) When the control group was exposed to 100 nmol/L,1 μmol/L,10μmol/L,20 μmol/LTCS,the frequency of c.elegans F1 body bent was (19.94±2.46)times/20 s,(15.13±1.99) times/20 s,(14.63±2.31)times/20 s,(14.69±1.96)times/20 s,(12.00±1.86)times/20 s,respectively,and the comparative differences between groups were statistically significant(F=25. 636,P<0. 01). (3) When the control group was exposed to 0,100 nmol/L,1 μmol/L,10 μmol/L,20 μmol/L TCS,the body sizes of the c. elegans F1 generation was (286.83±6.01)articles,(273.33±6.41)articles,(214.17±7.25)articles,(173.67±9.20)articles, (118. 50 ± 6. 98 ) articles, respectively, the brood size of the C. elegans F1 generation exposed to 100 nmol/L, 1μmol/L,10 μmol/L,20 μmol/L TCS levels,were reduced by 4. 71%,25. 60%,39. 45%,58. 67%,the ge-neration time of the c. elegans′F1 generation was shortened by 2. 14%-5. 38% in the TCS treatment groups compared with the control group(F=27. 520,P<0. 01). Conclusions After c. elegans exposure to TCS,locomotory behavior can be severe-ly affected,reproductive damage causes a decline in the number of brood size,and the speeding-up of the breeding rate is related to the concentration of TCS concentration-response.

Objective To investigate the eating problems of outpatient infants,preschool age children(1 to 7 years old) enrolled in the Department of Child Health Care,Nanjing Maternal and Child Health Hospital Affiliated to Nanjing Medical University,and to analyze its correlation with children's physical development,so as to establish strategies for preventing abnormal eating habit in children.Methods A toll of 2458 children met the criteria,and caregivers (mothers) completed the Children's Eating Behavior Questionnaires (CEBQ) in Department of Child Health Care,Nanjing Maternal and Child Health Hospital Affiliated to Nanjing Medical University were selected and the children's sociodemographic data and the morbidity of children eating problems were investigated.The correlation between children's body mass index(BMI) with children's eating problems was determined by using Chi-square test and multiple regression analysis.Results About 66.2％ (1627/2458 cases) had normal weight,and 10.8％ (257/2458 cases) and 8.5 ％ (210/2458 cases) were overweight (BMI ＞ P85-P95) or obese (BMI ≥ P95),respectively.The prevalence of eating behavior problems was detected during 25-36 months.For 1-to-7-year-old children,the highest detection rate of eating problems was inattention and eating at non-permanent locations,occupying 64.7％ (1590/2458 cases)and 50.5％ (1241/2458 cases),respectively;the prevalence rate of preferring to junk food was the lowest,accounting for 19.3％ (474/2458 cases).The children's eating problems had a high association with the children's BMI.Among them,children with eating problems,such as difficultly in accepting the varying food stuff[at the age of 12 month,odds ratio(OR)=11.50,95％ confidence interval(CI):1.84-72.16] and eating at non-permanent locations(at 25-36month,OR=1.77,95 ％ CI:1.11-2.83),were prone to be wasting away;children with eating problems,such as preferring to junk food (at 12 month,OR=5.08,95 ％ CI:1.43-18.00;13-18 month,OR=2.17,95 ％ CI:1.06-4.44),rarely eating vegetables or fruit (19-24 month,OR=4.06,95％CI:1.46-11.31) and inattention (12 month,OR=3.85,95 ％ CI:1.52-9.79),were associated with overweight or obesity (all P＜0.01).Conclusions There was a high prevalence of eating problems in children between 12-84 month(1-7 years old) in Nanjing.Improper children's eating behaviors can increase the risks of wasting away or children's overweight/obesity.