Objective To discuss the possible molecular mechanisms involved in the protective effects of Biifdobacterium on intestinal tissue of necrotizing enterocolitis (NEC) newborn rats. Methods Seventy-five newborn Sprague-Dawley rats (born within 2 h) were randomly divided into five groups, each group with 15 rats. Group A was the NEC model group, and the rats were fed lipopolysaccharide (LPS) and formula. Group B was the Biifdobacterium treatment group, and the rats were fed LPS and formula and Biifdobacterium micro-capsule. Group C was the artificial feeding control group, and the rats were fed formula. Group D was the Biifdobacterium control group, and the rats were fed formula and Biifdobacterium micro-capsule. Group E was the breastfeeding control group, and the rats were fed rat breast milk by mothers. LPS 30 mg/kg was administered by gavage once per day for 3 days. Bifidobacterium micro-capsules were given as 1×1010 colony forming units/ml by gavage with formula once per day. After fed for 72 h and fasted for 12 h, the five groups of rats were killed by decapitation. Morphological changes in the terminal ileum tissue were observed under a light microscope and intestinal injury was scored. The expression of Toll-like receptor (TLR) 2, TLR4, and nuclear transcription factor (NF)-κB p65 was detected by immunohistochemical methods. Kruskal-Wallis test, analysis of variance, corrected Chi-square test and Fisher's exact test were used for statistics. Results The morbidity of NEC in group A to E was 11/15, 4/15, 3/15, 2/15 and 0/15, respectively;the intestinal injury score in group A to E was 3.37±0.27, 1.53±0.44, 1.75±0.37, 0.92±0.39 and 0.30±0.18, respectively; the expression level of TLR2 in group A to E was 0.35±0.05, 0.30±0.03, 0.32±0.04, 0.30±0.02 and 0.29±0.03, respectively;the expression level of TLR4 in group A to E was 0.48±0.05, 0.34±0.03, 0.36±0.03, 0.37±0.04 and 0.35±0.02, respectively;the expression level of NF-κB p65 in group A to E was 0.43±0.03, 0.29±0.03, 0.35±0.02, 0.32±0.02 and 0.30±0.02, respectively. The differences in NEC morbidity, intestinal injury score, and the expression levels of TLR4, TLR2 and NF-κB p65 among the five groups were all statistically significant (χ2, H or F=23.863, 70.290, 8.803, 38.599 and 75.076, respectively, all P<0.05). The values in the NEC model group were all significantly higher than those in the other four groups (all P<0.05). The morbidity of NEC in the Biifdobacterium treatment group compared with the three control groups was not significantly different (all P > 0.05). The intestinal injury score in the Bifidobacterium treatment group was significantly higher than that in the Bifidobacterium control group and the breastfeeding control group (both P < 0.01), but was not significantly different to that in the artificial feeding control group (P > 0.05). The expression levels of TLR4 and NF-κB p65 in the Biifdobacterium treatment group were significantly lower than those in the artificial feeding control group and the Biifdobacterium control group (all P < 0.05), and were not significantly different to those in the breastfeeding control group (P>0.05). The expression level of TLR2 in the Biifdobacterium treatment group compared with the three control groups was not significantly different (all P > 0.05). Conclusions Biifdobacterium may inhibit pathogenic bacteria or regulate the negative feedback of TLR2 to reduce the expression of TLR2 and TLR4 in intestinal mucosa cells, inhibit the NF-κB pathway, attenuate the inflammatory reaction, and play a role in the prevention and control of NEC.

Objective To study the effect of bifidobacterium on intestinal tissue of necrotizing enterocolitis (NEC)in newborn rats and its regulation of Wnt/β-Catenin signal pathway.Methods Seventy -five newborn SD rats were randomly divided into 5 groups,and each group had 1 5 rats.Group A was artificial feeding control group;group B was NEC model group;group C was bifidobacterium treatment group;group D was artificial feeding +bifidobacterium control group;group E was rat breast feeding control group.The localization expression of Toll -like re-ceptor 4(TLR4)of ileocecal ileum tissue was detected by immunohistochemical detection,and also the equivalen-tileum tissues were detected for the contents of glycogen synthase kinase -3β(GSK3β)and β-Catenin expression by Wes-tern blot.Comparing the differences of these indicators between the groups,in addition,the data of TLR4,GSK3βandβ-Catenin were analyzed by Bivariate correlations.Results The levels of TLR4 in ileum tissue of 5 groups were 0.36 ±0.03,0.48 ±0.05,0.34 ±0.03,0.37 ±0.04,0.35 ±0.02.The levels of GSK3βin ileum tissue of 5 groups were 0.98 ±0.23,1 .48 ±0.42,0.99 ±0.20,0.56 ±0.1 7,0.60 ±0.1 5.The levels of β-Catenin in ileum tissue of 5 groups were 1 .48 ±0.22,0.64 ±0.55,1 .27 ±0.36,1 .72 ±0.51 ,1 .82 ±0.44.The levels of TLR4 and GSK3βin ileum tissue of group B were significantly increased compared with group E (P <0.05).The levels of β-Catenin sig-nificantly decreased compared with group E (P <0.05).The levels of TLR4 and GSK3βin ileum tissue of group C were significantly decreased compared with group B (P <0.05).The levels of β-Catenin significantly increased com-pared with group B (P <0.05).Negative correlation was observed between the levels of GSK3βand β-Catenin(r =-0.592,P <0.05),while positive correlation was observed between the levels of TLR4 and GSK3β(r =0.295,P <0.05),and negative correlation was observed between the levels of TLR4 and β-Catenin(r =-0.426,P <0.05). Conclusions Bifidobacterium has certain protective effect on the NEC newborn rat intestines,which can reduce the in-cidence of experimental NEC and the severity of intestinal injury.Its effect may be achieved by regulating the Wnt/β-Catenin signal pathway,which decreases the expression of the level of GSK3βand increases the level of repair fac-tor β-Catenin.

Objective:To investigate the protective effect and mechanism of microRNA-210 agonist (agomiR-210) on kidney in diabetic kidney disease (DKD) rats.Methods:Thirty-six 5-week-old male SD rats were divided into normal control (NC) group, agomiR-NC control group, agomiR-210 control group, DKD model group, DKD+agomiR-NC group and DKD+agomiR-210 group, with 6 rats in each group. Diabetic rats were established by a high-fat diet combined with intraperitoneal injection of streptozotocin (STZ), then were fed for 12 consecutive weeks to construct DKD model rats. During 2nd-4th week of continuous feeding, the rats in DKD+agomiR-210 group were injected with 20 nmol/kg agomiR-210 via tail vein twice a week. Blood glucose levels, 24 h urine albumin (Alb) and 24 h urine microalbumin (MAU) contents were measured regularly. At the end of the 12th week, the rats were sacrificed, and renal tissues were collected. The renal histopathological changes were assessed by HE, PAS and Masson staining methods. The mRNA and protein expression levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β and IL-6 in renal tissues were detected by RT-qPCR and Western blot. The distributions and expressions of α-smooth muscle actin (α-SMA), typeⅠ collagen (Col-Ⅰ), type Ⅳ collagen (Col-Ⅳ) and fibronectin (FN) in renal tissues were detected by immunohistochemical method. The protein expression levels of phospho(p)-Smad3 and p-NF-κB p65 in renal tissues were detected by Western blot and immunohistochemical methods.Results:Compared with DKD model group, the renal pathological damages in DKD+agomiR-210 group were improved, the blood glucose level, glycogen deposition and collagen accumulation were significantly decreased (all P<0.05), the urinary excretions of Alb and MAU were significantly reduced (all P<0.01), and the expressions of TNF-α, IL-1β, IL-6, α-SMA, Col-Ⅰ, Col-Ⅳ, FN, p-Smad3 and p-NF-κB p65 in renal tissues were significantly decreased (all P<0.01). Conclusion:AgomiR-210 can alleviate renal pathological changes and urinary Alb and MAU excretion in rats with DKD, which may be related to its inhibition of Smad3 and NF-κB activity.

Objective To study the relationship between serum neutrophil gelatinase-associated lipocalin (NGAL),kerbs von lungren 6 antigen (KL-6) and bronchopulmonary dysplasia (BPD) in preterm infants.Method From Jan.2015 to Dec.2015,preterm infants admitted to NICU of Guangzhou Women and Childrem's Medical Center with gestational age less than 32 weeks and birth weight less than 1 500 g were enrolled.The serum levels of procalcitonin (PCT),NGAL and KL-6 protein were detected at 24 h,7 d and 14 d after birth.At the 28 d after birth,according to the presence of BPD or not,the infants were assigned into BPD group and non-BPD group.The differences of the serum levels of PCT,NGAL and KL-6 between the two groups were compared.Result A total of 55 cases were included in the study (BPD group 20 cases,non-BPD group 35 cases).No significant differences existed in gender,birth weight and gestational age between the two groups (P ＞ 0.05).The incidence of respiratory distress syndrome were siginificantly higher in the BPD group (P ＜ 0.05) and the duration of mechanical ventilation and oxygen therapy were siginifantly longer in the BPD group (P ＜ 0.05).No significant difference between the two groups in the level of PCT at 24 h after birth (P ＞ 0.05).The levels of serum PCT at 7 d and 14 d after birth in BPD group were significantly higher than non-BPD group [7 d:(1.5 ± 1.7) ng/ml vs.(0.4 ± 0.2)ng/ml,14 d:(0.8 ± 0.7) ng/ml vs.(0.2 ± 0.1) ng/ml] (P ＜ 0.001).The levels of serum NGAL at 24 h,7 d and 14 d were significantly higher than non-BPD group [24 h:(1.6 ± 0.3) pg/ml vs.(0.8 ±0.2) pg/ml,7 d:(2.3 ±0.5) pg/ml vs.(0.7 ±0.2) pg/ml,14 d:(2.5 ±0.3) pg/ml vs.(0.8 ±0.2)pg/ml] (P ＜0.001).The levels of serum NGAL at 7 d and 14 d after birth in BPD group were significantly higher than 24 h in BPD group (P ＜ 0.05).No significant difference between KL-6 at 24 h after birth in BPD group and non-BPD group (P ＞0.05).The levels of serum KL-6 at 7 d and 14 d after birth in BPD group were significantly higher than non-BPD group [7 d:(1.2 ± 0.2) ng/ml vs.(0.8 ± 0.1) ng/ml,14 d:(1.3 ±0.2) ng/ml vs.(0.8 ±0.9) ng/ml] (P ＜0.001).The level of serum KL-6 at 7 d and 14 d after birth in BPD group were significantly higher than 24 h in BPD group (P ＜ 0.05).Conclusion Respiratory distress syndrome and prolonged mechanical ventilation were risk factors of BPD.The rising of serum NGAL and KL-6 early after birth might be involved in the development of BPD,which had predictive value of BPD.

Objective: To analyze the clinical characteristics of community-acquired pneumonia (CAP) in children under five years of age and analyze the safety and efficiency of nasal continuous positive airway pressure (NCPAP) ventilation for CAP in this population. Method: This was a prospective multicenter study. Children who were admitted to these six centers with CAP and met the NCPAP ventilation indications, aged from 29 d to 5 years, were continuously included during November 2013 to October 2015. The baseline data were collected and NCPAP ventilation were then followed up by operation standards, and the vital signs and arterial blood gas change at special time points were observed and recorded. Any side effect associated with NCPAP were recorded. For categorical variables, comparisons were performed using Fisher test. Rank-sum test and t test were performed respectively for abnormal and normal distribution continuous variables. The variables pre-NCPAP and post-NCPAP were analyzed by repeated measures ANOVA analysis. Result: Totally 145 children were included, and 13 children were excluded due to incomplete data. One hundred and two children(77.3%)were ≤12 months; 91 children (68.9%) were from rural area. NCPAP ventilation was effective in 123 children, with a response rate of 93.2%, were all discharged with a better condition; it was ineffective in 9 children(6.8%), and they were all intubated and went on mechanical ventilation, 5 were discharged with a better condition, and 4 died after gaving up treatment. The gender, age, body weight, residence, main symptoms, main signs, imaging diagnosis, medications, partial pressure of oxygen(PaO₂), breath and heart rate before NCPAP treatment of two groups had no significant differences(allP>0.05). The rates of combining underlying diseases, trouble with feeding and cyanosis, and the partial pressure of carbon dioxide(PaCO₂ ) before NCPAP ventilation were higher in NCPAP ineffective group ((59±11 )vs.( 49±11) mmHg, 1 mmHg=0.133 kPa, t=-2.597, P=0.028); while the PaO₂/fraction of inspiration O₂ (FiO₂ ) before NCPAP was lower((150±37) vs. (207±63) mmHg, t=2.697, P=0.008). The breathing, heart rate and PaCO₂ of NCPAP effective group decreased significantly, while the PaO₂ and PaO₂/FiO₂ increased significantly after 2, 8, 24 h of NCPAP ventilation(all P=0.000). PaCO₂ in children with hypercapnia before NCPAP ventilation in NCPAP effective group decreased significantly ((48±9), (47±12), (45±11)vs.(58±7)mmHg, all P=0.000). All children tolerated well to NCPAP ventilation, and there were no severe side effects or complications associated with NCPAP ventilation. Conclusion NCPAP ventilation is safe and effectively improved the oxygenation and hypercapnia in infants with CAP. But it may not work well in children with underlying diseases, manifest as difficulty in feeding/cyanosis and extremely high PaCO₂ or low PaO₂/FiO₂, and they may need early intubation.