Neonatal necrotizing enterocolitis (NEC) is still a disease with high mortality and morbidity in neonates.The pathogenesis of NEC is not quite clear.Inflammatory cascade,innate immune response and intestinal blood flow are thought to play significant roles in the machinism of NEC.Single nuleotide polymorhism (SNP) is characterized by single nucleotide mutation in the genecome consequences.They may alter the susceptibility to some kinds of diseases by changing the efficiency of transcription or the structure of protein coded by the related gene.This review summarizes the progress of the research on genetic polymorphisms in neonatal necrotizing enterocolitis.

In recent years,clinical immunology has an increasing variety of prominent applications in the field of chronic hepatitis B infection.On one hand,the discovery and identification of the mechanisms of pathogen recognition by the innate immune system,novel immune cell subpopulations and immunoregulatory pathways further enriched the immunological pathogenesis of chronic hepatitis B.On the other hand,the development of new immunotherapy strategies aimed at reconstructing the antiviral immunity,including agonists of Toll like receptors,immune cell therapies and therapeutic vaccines,have provided several new targets for the optimization of antiviral therapy strategies and brought new opportunities for patients of chronic hepatitis B to obtain HBsAg clearance and clinical cure.

Objective To investigate the efficacy observation of transcranial direct current stimulation (tDCS)for improving the attention in patients with infarction in basal ganglia area. Methods Sixty consecutive patients with basal ganglia infarction admitted to the Department of Rehabilitation Medicine,the First Affiliated Hospital of Nanchang University from May 2015 to May 2016 were enrolled. They were randomly divided into either a control group or a test group according to the random number table (n = 30 in each group). The patients in both groups received routine rehabilitation training,and those in the test group received tDCS therapy synchronously,and the control group received the sham tDCS stimulation. The evaluations and analyses were conducted with the Montreal cognitive assessment (MoCA),d2 test of attention,and event-related potential-P300 (ERP-P300),respectively in all patients before intervention and 4 weeks after intervention,and they were compared between the groups. Results There was no significant difference before intervention between the two groups (all P > 0. 05). Compared with before intervention,the ERP-P300 latencies were reduced,the amplitudes were increased after intervention in the patients of the test group and the control group (all P < 0. 05). The MoCA total scores (the test group:22. 7 ± 2. 7 vs. 15. 5 ±
2. 4;the control group:17. 2 ±2. 5 vs. 15. 6 ±2. 3),attention dimension scores (the test group:4. 6 ± 1. 2 vs. 2. 4 ± 1. 6;the control group:3. 6 ± 1. 5 vs. 2. 5 ± 1. 5),and the total completion of d2 attention test task, total scores,and concentration degree scores (the test group:295 ± 31 vs. 250 ± 45,279 ± 38 vs. 223 ± 52, 97 ± 22 vs. 75 ± 25;the control group:276 ± 33 vs. 247 ± 45,257 ± 39 vs. 211 ± 56,84 ± 23 vs. 71 ± 24) were all increased (all P < 0. 05),and all the indexes of the test group were better than those of the control group (all P < 0. 05). Conclusion tDCS contributes to the improvement of the attention in patients with infarction in the internal capsule-basal ganglia region.

Objective To study the feasibility and invasiveness of hand-assisted laparoscopic hepatectomy(HALH) for liver cancer.Methods Forty patients undergoing hepatectomy for liver cancer were randomly divided into HALH group and open hepatectomy(OH) group.Data of patients of two groups,Which included operating time,intraoperative blood loss,length of incision,postoperative flatus time,hospital stay,complications and C-reactive protein(CRP) were compared.Results The mean intraoperative blood loss,length of incision,postoperative flatus time,hospital stay and CRP in HALH group were significantly less than that in OH group;but there was no significant difference in operating time,or complication and recurrence rate.Conclusions HALH for liver cancer is less traumatic,and achieves faster patient recovery.It is feasible and safe in selected patients.

Objective To investigate the effect of heparin-binding epidermal growth factor-like growth factor (HB-EGF) on mitochondrial pathway of apoptosis in rats with neonatal necrotizing enterocolitis (NEC).Methods Sprague-Dawley neonatal rats were randomly divided into three groups with ten in each.NEC group rats were formula fed,and hypoxia exposed by 100％ N2 for 90 s and cold stress at 4 ℃ for 10 min twice a day for three days.Additionally,rats in HB-EGF group received HB-EGF 800μg/kg by gavage four times a day for three days.Rats in control group were given breast milk feeding for three days without any interventions.Seventy-two hours after born,all neonatal rats were sacrificed after fasting for 12 h,from which the terminal ileum was removed.HE-staining was done for histologic evaluation.Mitochondrial ultrastructure was observed under electron microscopy.Cytochrome C was detected by immunohistochemical analysis and apoptosis inducing factor (AIF) and apoptotic protease activating factor-1 (APAF-1) were measured by Western blot.Analysis of variance and q-test were used to compare the difference among groups.Results (1) The incidence of NEC in HB-EGF group was lower than that in NEC group (2/10 vs 9/10,x2 =7.27,P＜0.01).(2) In NEC group,mitochondria in epithelial cells and muscle cells of intestine were significantly swelling,appearing many electron-lucent zones in matrix.Ultrastructure of mitochondria were severely damaged.In HB-EGF group,mitochondria were less swelling and showed milder damage than those in NEC group.(3) The expression of cytochrome C in ileal tissue in NEC group was higher than that in control group (0.030±0.018 vs 0.002±0.001,q=6.15,P＜0.01).The expression of cytoehrome C in ileal tissue in HB-EGF group was lower than that in NEC group (0.014±0.018 vs 0.030±0.018,q=3.53,P＜0.05).The expression of APAF-1 and AIF in NEC group was higher than those in control group (1.364±0.299 vs 0.215±0.033,q=15.31,P＜0.05;0.181±0.050 vs0.127±0.045,q=3.71,P＜0.05).Compared to NEC group,the expression of APAF-1 was lower (0.455±0.123 vs 1.364±0.299,q=4.04,P＜0.05) and the expression of AIF was higher (0.289±0.045 vs 0.181±0.050,q=7.32,P＜0.05) in HB-EGF group.Conclusions HB-EGF could reduce the incidence of NEC in neonatal rats by inhibiting the mitochondrial pathway related apoptosis through down regulation of APAF-1.

Objective To detect the effects of bifidobacterium or bifidobacterium cultured supernatant on the mRNA expression of tumor necrosis factor receptor-associated factor 6 (TRAF6),glycogen synthase kinase-3β (GSK-3β) and the miRNA-146a in rat small intestinal epithelial cell(IEC-6) induced by lipopolysaccharide (LPS).Methods IEC-6 in logarithmic phase were randomly divided into LPS group,cultured supernatant group and inactivated bacteria group.All the 3 groups were exposed to 5 mg/L LPS for 5 hours,and then 1 mL sterile saline was added in LPS group and culturing continued for 24 hours ; 1 mL bifidobacterium cultured supernatant was added in cultured supernatant group and culturing continued for 24 hours;1 mL inactivated bifidobacterium 1 x 1010 CFU/L added in inactivated bacteria group and continued culturing for 24 hours.The mRNA expressions of TRAF6,GSK-3 β and miRNA-146a were detected by reverse transcription-polymerase chain reaction (RT-PCR).Results The level of TRAF6,GSK-3 β of culture supematant group (0.72 ± 0.05,0.46 ± 0.14) were all lower than LPS group (1.01 ± 0.14,1.02 ± 0.25),but the level of miRNA-146a(3.05 ± 0.40) was higher than that in LPS group(1.01 ± 0.12),and there were significant differences between them (t =5.278,6.316,13.218,P =0.000).The level of GSK-3 β of inactivated bacteria group(0.59 ±0.13) was significantly lower than that in LPS group(t =4.837,P =0.000).The levels of TRAF6 and miRNA-146a of inactivated bacteria group(1.05 ±0.11,0.78 ±0.22) had no significant differences with LPS group (t =0.732,1.463,P ＞ 0.05).The level of TRAF6 of cultured supernatant group was lower than that in inactivated bacteria group,and the level of miRNA-146a was higher than that in inactivated bacteria group,and there were significant differences between 2 groups (t =6.009,14.687,P =0.000).Conclusions Bifidobacterium cultured supernatant and inactivated bacteria both have certain protective effect on the IEC-6 induced by LPS.One of the protective mechanisms of bifidobacterium cultured supernatant may be achieved by elevating the expression of miRNA-146a,and decreasing the levels of inflammation related factor TRAF6 and damage related factor GSK-3β.The protective effects of inactivated bifidobacterium may be achieved by decreasing the level of damage related factor GSK-3β.

ObjectiveTo analyze the differential expression of microRNA (miRNA) and its significance in patients with neonatal necrotizing enterocolitis (NEC).MethodsTwenty-five patients diagnosed with NEC with Bell stage≥Ⅱ, and 25 non-NEC patients as control group admitted to Guangzhou Women and Children's Medical Center between October 2014 and November 2015 were collected. White blood cells were extracted from the peripheral blood. Five samples were selected randomly each from NEC group and control group, and sequenced by second-generation Illumina high-throughput sequencing, screened for differentially expressed miRNA and analyzed for target genes prediction and biological function. The rest samples of the two groups were detected by real-time fluorescent quantitative polymerase chain reaction technology (RT-qPCR), the results were used to validate the results of high-throughput sequencing. Differentially expressed miRNA in the two groups of data was analyzed using DEGseq software.P<0.05 was considered statistically significant.P<0.01,q<0.001 and丨Log2 Ratio丨≥1 were taken as criteria for screening the differential expression. The differential expressions of miRNA in NEC group and control group were analyzed by cluster analysis using MeV4.6 software.ResultsA total of 482 miRNAs were differentially expressed in the two groups, with significant difference (P<0.05). Among them, 126 were known miRNAs with significantly differential expression in the two groups, with 58 being up-regulated and 68 being down-regulated. The results of up-regulated miRNAs (hsa-miR-223-5p,-183-3p,-222-5p) and down-regulated miRNAs (hsa-miR-23b-5p,-150-5p,-146a-3p,-1298-5p) were confirmed to be consistent with the results of sequencing. Bioinformatics analysis showed that the target genes with differential miRNA expression mainly involved Toll-like receptor signal transduction pathway, mitogen-activated protein kinase pathway, JAK-STAT and other signal transduction pathways.ConclusionsThere are significantly differential expressions of miRNAs in peripheral white blood cells of NEC neonates. These miRNs may be involved in the occurrence and development of NEC via adjusting different target genes to regulate the signal pathway.

Objective To investigate the relationship between myeloid differentiation (MD-2) gene polymorphisms and necrotizing enterocolitis (NEC) in neonates.Methods A gene-sequencing method was used to re-sequence the exons and the promoter functional polymorphism region (rs11465996) of MD-2 gene of 42 NEC neonates admitted in neonatal intensive care unit of Women and Children's Medical Center,Guangzhou Medical University from June 1,2011 to May 31,2012.These functional polymorphism loci were compared with 83 non NEC cases.The Chi square test was used for statistical analysis.Results No polymorphism was detected in the exons of MD-2 gene in any of the 42 cases of NEC.The C-1625G polymorphism [rs11465996 (C＞G)] was identified in both the NEC and control groups,and there were two genotypes,C/C and C/G.The frequency ofC/G genotype in the NEC group (38.1％,16/42) did not differ significantly compared to that in the control group (30.1％,25/83) (x2=0.805,P=0.370).However,the frequency of C/G genotype in severe NEC cases (operation group) (55.0％,11/20) was significantly higher than that in the control group (x2=4.388,P=0.036).Among the NEC group,the frequency of C/G genotype in operation cases and term infants was higher than that of the non-operation cases and preterm infants,although the differences were not significant (x2=3.343,P=0.067; xx2=0.913,P=0.339).Conclusions The polymorphisms in the exons of MD-2 gene are not associated with the development of NEC.The rs 1 1465996 polymorphism (G allele) in the promoter region may be related to the severity of NEC.

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 investigate the effects of lipopolysaccharides (LPS) in different concentrations on the proliferation and interleukin(IL)-6,IL-1 β and tumor necrosis factor-α(TNF-o) secretion of intestinal epithelial cells (IEC-6) of rats in vitro.Methods IEC-6 of rats were divided into normal group (0 mg/L,group A),0.1 mg/L group (group B),0.5 mg/L group (group C),1.0 mg/L group (group D),5.0 mg/L group (group E) and 10.0 mg/L group(group F).Different groups cells were exposed to LPS with different concentrations for 3 h,5 h and 7 h.Thiazolyl blue(MTT) was performed to investigate the proliferation of IEC-6.The concentrations of IL-6,IL-1 β and TNF-α in culture supernatant were detected by enzyme linked immunosorbent assay(ELISA).Results The proliferation rate of IEC-6 was gradually lower while the concentration of LPS increased.After co-culture with LPS 3h and 5 h,the proliferation rates of group B,group C,group D,group E and group F had no significant difference with those of group A (all P ＞ 0.05);after co-culture with LPS 7 h,the proliferation rates of group B,group C,and group D had no significant difference with those of group A (all P ＞ 0.05);the proliferation rates of group E and group F had significant difference with those of group A(t =4.216,P =0.014;t =14.991,P =0.000).The proliferation rates of group E and group F were lower after co-culture with LPS 5 h than 7 h,and there were significant differences (t =2.576,P =0.033;t =2.975,P =0.018);but there was no significant differences between group E and group A after co-culture with LPS 5 h (P ＞ 0.05).Group B,group C,group D,group E and group F all had a significant higher level of IL-6 than group A after co-culture with LPS 3 h,5 h and 7 h(all P ＜0.01).In addition,group E had the highest level of IL-6 at all time points.And the peak level of IL-6 rose after co-culture with LPS 5 h.The TNF-α level and IL-1 β level of group B,group C,group D,group E and group F all had no significant differences than that of group A after co-culture with LPS 3 h,5 h and 7 h (all P ＞ 0.05).Conclusions In a certain concentration,incubation time range,the proliferation rates of IEC-6 cells were gradually lower while the concentration of LPS increased.Co-cultured IEC-6 cells with LPS(0-10.0 mg/L) can stimulate them secrete to IL-6.The highest level of IL-6 was of group E after 5 h co-culture.LPS had no effects on TNF-α and IL-1 β level of IEC-6 cells cultural supernatant.So 5.0 mg/L concentration of LPS stimulating IEC-6 cells for 5 h can be chosen to build the IEC-6 inflammatory models.