Extrauterine growth restriction is a major clinical problem for very low and extremely low birth weight infants.They need the rapid accumulation of protein and energy after born;otherwise it will lead to an increase in nutritional deficiencies,malnutrition and neurological problems.The ideal nutritional strategies is to provide parenteral nutrition after birth,amino acids and lipids are initially provided at a minimum of 2.0 g/(kg · d) and 1.0 g/(kg — d),which advance to 3.5-4.0 g/(kg · d) and 3.0 g/(kg · d),keep the appropriate protein/energy ratio.The appropriate vitamin,minerals and elements should be provided.Enteral feeding should be initiated immediately about 24 h after birth,minimal feeding [10-20 mL/(kg · d)] should last 3-5 days.Human milk should gradually increase to 75-120 mL/(kg · d) and add human milk fortifier.Parenteral nutrition is stopped when human milk fed to 120 mL/(kg · d).

Objectives:To observe the role of combined use of early micro feeding and intravenous nutrition in low birth weight(LBW) infants. Methods:Fifty four cases of LBW infants were randomly divided into two groups.Early micro feeding and intravenous nutrition were given in one group(EF & IN group) and another group(Control group) was given only with intravenous nutrition.The times of intravenous nutritional support requirement and hospital stay,the increase in body weight and the changes in serum bilirubin,lipids and creatinine were compared between the two groups. Results:The times of intravenous nutritional support and hospital stay were shortened and the body weight was increased in EF & IN group. The levels of serum bilirubin and creatinine and the serial concentrations of lipids on the day 7 and 14 after birth were significantly lower than those in control group. Conclusions:The combination of early,micro feeding and intravenous nutrition is superior to the only use of intravenous nutrition in shortrenning the critical course,increasing the body weight,beginning the oral intake and decreasing the possible injuries from total parenteral nutrition.

Objective To evaluate the safety and effects of feeding very low birth weight infants and extremely low birth weight infants with human donors' milk in China.Methods One hundred and ninety-nine very low birth weight infants and extremely low birth weight infants (VLBWI/ELBWI) admitted to NICU in Nanjing Maternity and Child Health Care Hospital were analyzed retrospectively (from August 1,2013 to July 31,2014) one year before and one year after the establishment of human donors' milk bank.Only 1 infant's parents(1/199 cases) refused to use donor milk,and 198 infants were fed with donors' milk since their own mothers' milk was unavailable.One hundred and forty-eight infants admitted to NICU from August 1,2012 to July 31,2013 were chosen as the control group,who were fed with formula milk since their own mothers' milk was unavailable.The mortality,incidence of necrotizing enterocolitis (NEC) and sepsis,time to full enteral feeding and hospital stay between 2 groups were analyzed.Results One year before the establishment of breast milk bank,there were 148 cases of very low or extremely low birth weight infants receiving mixed feeding,and 16 cases of them had neonatal necrotizing enterocolitis,the incidence rate was 10.81％.Since the establishment of human milk bank,187 women donated breast milk in the first year and 260 555 mL of human milk were collected and delivered.Only 1 infant (1/199 cases) refused to use donor milk,and 198 infants were fed with donors' milk since their own mothers' milk was unavailable.Nine babies with VLBWI/ELBWI were diagnosed as NEC in donor milk feeding group,and 16 cases were diagnosed as NEC in formula-feeding group.The incidence rate of NEC was lower in the donor-milk-feeding group (4.52％ vs 10.81％,x2 =5.02,P ＜ 0.05).Donor-milk-feeding group reached full enteral feeding[(16.4 ± 5.3) d vs (18.2 ± 6.2) d,t =2.84,P ＜ 0.05] and had shorter period of hospital stay[(35.1 ± 9.5) d vs (37.3 ± 10.4) d,t =2.05,P ＜ 0.05] compared with formula feeding group.The mortality and incidence of sepsis had no difference between 2 groups (all P ＞ 0.05).Conclusions Donors' breast milk is associated with a lower risk of NEC,sooner full enteral feeding and shorter hospital stay.Further research is needed to confirm these findings and measure the growth and long term neurodevelopment effects of donors' breast milk.

The Human Milk Banking Association of North America(HMBANA) is a professional association for supporters of non-profit donor human milk banking.HMBANA develops guidelines for donor human milk banking practices.The construction of China's human milk bank has just begun in few areas in 2013.To further promote the development and implementation of donor human milk banking in our country,this review introduce the guideline for establishment and operation of a donor human milk bank which drafted by HMBANA in 2013.The guideline can provide the basis for the development of human milk bank in our country.

Objective To investigate the effect of S100A4 silencing on tumor related gene COX-2,bcl-2,Surviving,MMP-9 mRNA expressions of pancreatic cancer BxPC-3,AsPC-1 cells,and explore their relationship.Methods Small interfering RNA interfering S100A4 gene (siRNA-S100A4) was applied to transfect human pancreatic cancer BxPC-3,AsPC-1 cells,and nonhomologous siRNA-C was used as negative control,and cells without transfection were used as control group.The expressions of S100A4,COX-2,Survivin,MMP-9,bcl-2 mRNA after interference were detected by using RT-PCR.Results S100A mRNA expressions of BxPC-3's control group,siRNA-C group,siRNA-S100A4 group were 0.661 ± 0.023,0.659 ± 0.043,0.379 ± 0.039,and expressions of COX-2 mRNA were 0.760 ± 0.026,0.830 ± 0.017,0.443 ±0.006,and expressions of Survivin mRNA were 0.948 ± 0.049,0.909± 0.081,0.068 ± 0.006,and expressions of bcl-2 mRNA were 0.462 ±0.018,0.421 ±0.049,0.184 ±0.025,and expressions of MMP-9 mRNA were 0.813 ± 0.008,0.908 ± 0.063,0.246 ± 0.027.S100A mRNA expressions of AsPC-I's control group,siRNA-C group,siRNA-S100A4 group were 0.641 ± 0.042,0.626-± 0.053,0.320 ± 0.081,and expressions of COX-2 mRNA were 0.727 ± 0.021,0.743 ± 0.025,0.560 ± 0.035,and expressions of Survivin mRNA were 0.994 ± 0.032,0.984 ± 0.049,0.063 ± 0.005,and expressions of bcl-2 mRNA were 0.458 ±0.004,0.537 ± 0.046,0.181 ± 0.007; and expressions of MMP-9 mRNA were 0.698 ± 0.011,0.718 ± 0.073,0.199± 0.013.The expressions of S100A,COX-2,Survivin,bcl-2,MMP-9 mRNA in groups with siRNA-S100A4 transfection were significantly lower than those of siRNA-C group and control group (P ＜0.01),but the difference between siRNA-C group and control group was not statistically significant.Conclusions S100A4 plays a role in the pathogenesis of pancreatic cancer through up-regulation of COX-2,Survivin,bcl-2,MMP-9 expressions.

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 risk of neonatal hyperbilirubinemia by transcutaneous bilirubin (TcB) nomograms and clinical risk factors.Methods Healthy term and late-preterm newborns (≥ 35 gestational weeks,and birth weight ≥ 2 000 g) born in Guizhou Maternal and Child Care Hospital between January 1,2013 and December 31,2013,were included.TcB levels were continuously recorded within 168 hours after birth.The value of hour-specific TcB nomogram combined with receiver operating characteristic (ROC)curves and Logistic regression model for predicting risk of hyperbilirubinemia was evaluated.Pearson's Chisquare test was also used for statistical analysis.Results A total of 5 250 cases were enrolled.TcB increased rapidly in the first 40 hours after birth,slowly increased between 40 to 96 hours,and reached a high level after 96 hours.Among them,the 95th percentile TcB stablized at 96 hours after birth.The 40th,75th and 95th percentile TcB peak levels were 173,217 and 248 μmol/L.Among the 5 250 neonates,there were 277 cases (5.3％) in the high-risk zone within 72 hours.The positive predictive value (PPV) was 22.02％;1 087 cases (20.7％) and 1 854 cases (35.3％) were in the medium-high risk and medium-low risk zones along with the PPV of 10.58％and 3.72％,respectively.There were 2 032 cases (38.7％) in the low-risk zone with the PPV of 1.38％.Multivariate analysis showed that the TcB high-risk zone after 72 hours was associated with gestational age,delivery mode,feeding mode and TcB level of risk zones within 72 hours.Compared to those born at ≥ 40 gestational weeks,those born at ≥ 37-＜40 gestational weeks were more likely in the TcB high-risk zone after 72 hours (OR=1.80,95％CI:1.29-2.51).The likelihood was reduced by 42％ among neonates born with cesarean section compared to those delivered vaginally in term of the TcB high-risk zone after 72 hours.Infants who received mixed feeding were less likely to be in the TcB high-risk zone after 72 hours when compared to breastfed infants (OR=0.51,95％CI:0.29-0.88).With the reduction of the high-risk zone level within 72 hours,the likelihood in the TcB high-risk zone after 72 hours was also decreased.ROC curve showed that the area under the curve (AUC) for predicting hyperbilirubinemia was 0.75 and its 95％CI was 0.72-0.78,with a sensitivity of 90.00％ and specificity of 40.00％.The AUC of a combination of predictive results obtained by the Logistic regression model with significant variables in univariate analysis and high-risk zone after 72 hours was 0.66,and its 95％CI was 0.62-0.69.AUC estimated by Logistic regression model according to the TcB levels of risk zones within 72 hours combining with clinical risk factors was 0.79,and its 95％CI was 0.76-0.82 (P＜0.01).Conclusions Hour-specific TcB nomograms of newborns in our hospital have been obtained,which facilitates the prediction and early intervention of neonatal hyperbilirubinemia.

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 analyze Holder pasteurization,temperature and duration of frozen,duration of continuous breast milk pumping and types of pumping tube on macronutrients and total energy in breast milk and the circadian variation of macronutrients and energy content of breast milk.Methods Two hundred and two breast milk samples were collected from healthy mothers of 188 infants who were hospitalized in Department of Pediatrics,Obstetrics and Gynecology Hospital Affiliated to Nanjing Medical University from April 2016 to December 2016.Fat,protein,carbohydrate and energy contents of the breast milk samples were detected both before and after various processes to analyze the influence of Holder pasteurization,frozen storage conditions (temperature and time),duration of continuous pumping and types of pumping tube and to observe the variations within a single day.Paired t-test,repeated-measures analysis of variance,one-way analysis of variance and LSD-t test were used for statistical analysis.Results The contents of fat,protein and energy in breast milk were significantly decreased [(3.69 ± 0.98) vs (4.22 ± 1.00) g/dl,t=9.405;(2.12 ± 0.34) vs (2.21 ± 0.34) g/dl,t=4.785;(69.77±9.35) vs (74.94±9.53) kcal/dl (1 kcal=4.184 kJ),t=9.470],while the carbohydrate content was increased [(6.66±0.17) vs (6.62±0.17) g/dl,t=-3.442] after Holder pasteurization comparing with those before (all P＜0.01).Breast milk frozen at =80 ℃ had higher protein content than that frozen at-20 ℃ (Ftemperature=4.990,Ptemperature=0.027).Breast milk stored at-20 ℃ for 7,15,30,60 and 90 days showed significantly decreased fat,protein and energy contents as compared with those in fresh breast milk,while the carbohydrate contents in the breast milk samples stored at-20 ℃ for 15,30 and 60 days were significantly increased and that kept for 90 days were lower than those at 30 and 60 days (LSD-t test,P＜0.05).Compared with fresh breast milk,breast milk stored at-80 ℃ showed significantly decreased fat,protein and energy contents at 60 d and 90 d,but increased carbohydrate content at 60 d (LSD-t test,P＜0.05).Compared with fresh breast milk,breast milk which was pumped at 1,2 or 3 h through gastric tube (size 6,8 or 10) or transfusion connecting pipe had lower fat content (LSD-t test,P＜0.05).Compared with fresh milk,breast milk had lower energy content on condition that it was pumped through gastric tube size 6 at 2 or 3 h or through gastric tube size 8 or 10 or transfusion connecting pipe at 1,2 or 3 h (LSD-t test,P＜0.05).Breast milk secreted from 12:01 to 15:00 had higher fat and energy contents [(5.16±0.93) g/dl and (86.96±9.35) kcal/dl] than that secreted from 00:01 to 03:00[(4.54±0.75) g/dl and (80.77±7.75) kcal/dl],from 03:01 to 06:00 [(4.61 ±0.75) g/dl and (81.44±7.69) kcal/dl] or from 21:01 to 24:00 [(4.71 ± 0.88) g/dl and (82.44± 8.82) kcal/dl].Moreover,breast milk secreted from 09:01 to 12:00 had higher fat and energy contents [(5.01 ± 0.94) g/dl and (85.31 ± 9.62) kcal/dl] than that secreted from 00:01 to 03:00,and that secreted from 09:01 to 12:00 had higher fat contents than that secreted from 03:01 to 06:00 (LSD-t test,P＜0.05).Conclusions Fresh breast milk is better than processed milk to meet the infantile need for growth and development.

Objective To explore the effect of shRNA silenced aromatic hydrocarbon receptor (AHR) on WNT signaling path-way during the differentiation of P19 cells into cardiac myocytes. Methods The eukaryotic expression vector of mouse AHR gene was designed and constructed. The interference plasmid was transfected into P19 cell and the positive stains to AHR gene silencing were screened by G418. The mRNA expression of important genes GSK3βandβ-catenin were evaluated by real-time fluorescent quantita-tive PCR during the differentiation of P19 cells. Results The constructed AHR-shRNA plasmid significantly inhibited the expression of AHR gene. Along with the differentiation of P19 cell into cardiac myocytes, in the interference group the expression ofβ-catenin gene was lower whereas the expression of GSK3βgene was elevated than those of control group with significant differences (all P<0.01). Conclusions The interference of AHR gene expression can regulate WNT signaling pathway in the development of heart.