Objective To explore the clinical features and risk factors of poor prognosis in neonatal necrotizing enterocolitis(NEC).Methods A retrospective study was carried out in the infants with NEC admitted to 6 cooperative hospitals in Guangdong Province between January 2005 and December 2014.The clinical features and risk factors of poor prognosis in preterm and full-term infants diagnosed NEC,early onset and late onset NEC were analyzed.Results A total of 449 cases who met the criteria were admitted during the study time.The mortality was 23.6％ (106/449 cases),of which the preterm group was 24.6％ (58/238 cases) while the full-term group was 22.7％ (48/211 cases),the early onset group was 22.1％ (45/204 cases) while the late onset group was 24.3％ (57/235 cases).The median number of NEC onset in preterm group was 11 d after birth while the number of the full-term group was 6 d.Full-term infants who diagnosed NEC were more likely to manifest themselves as abdominal distension (52.1％ vs.42.0％,x2 =4.597,P =0.032),vomiting(36.5％ vs.17.2％,x2 =21.428,P =0.000) and bloody stool(30.3％ vs.21.4％,x2 =4.653,P =0.031);but in the onset of NEC,preterm infants more likely to have feeding intolerance (21.0％ vs.12.8％,x2=5.309,P =0.021).The early onset group of full-term NEC was much common in twins or multiplets(9.4％ vs.1.1％,x2 =6.226,P =0.013),which rate of surgical therapy was much higher (41.0％ vs.27.0％,P =0.036) and the breast-feeding rate before NEC was lower than the late onset group(14.5％ vs.32.6％,x2 =9.500,P =0.002),the differences were statistically significant.The gestational age and birth weight were bigger in the early onset group of preterm NEC[(33.8 ±2.5) weeks vs.(32.2 ±2.8) weeks,t =4.261,P =0.000;(2.1 ±0.5) kg vs.(1.7 ± 0.5) kg,t =4.735,P =0.000)],but length of stay was shorter than the late onset group (18.0 d vs.26.5 d,P =0.000).Logistic regression analysis showed that the risk factors of poor prognosis of full-term NEC were shock,peritonitis and sepsis;while risk factors of poor prognosis of preterm NEC were small for gestational age infant,pulmonary hemorrhage,shock,intestinal perforation and sepsis;the risk factors of poor prognosis of the early onset group of full-term NEC was shock;while those of the late onset group were shock and peritonitis;the risk factors of poor prognosis in the early onset group of preterm NEC were shock and sepsis,while those in the late onset group were pulmonary hemorrhage,shock,intestinal perforation and sepsis.Conclusions Compared to the preterm NEC,the onset time of full-term NEC was earlier and the clinical manifestations were more typical.Early identification and management of shock,peritonitis,intestinal perforation,sepsis and pulmonary hemorrhage can reduce the risk of poor prognosis of neonate NEC.

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 study the composition of gut microbiome in neonates with severe bilinebinemia (serum total bilirubin ＞ 342 μmol/L),and to explore the relationship between gut microbiome and bilirubin brain injury.Methods A prospective study was conducted.The neonates with serum total bilirubin ＞ 342 μmol/L from September 2016 to March 2017 in Guangzhou Women and Children's Medical Center,Guangzhou Medical University,were enrolled in the study and 16S rDNA sequence analysis technology was used to detect the composition of gut microbiome in all subjects.According to the results of brain magnetic resonance imaging (MRI),brain stem auditory evoked potential (BAEP) and clinical manifestations,the subjects were divided into the brain injury group (26 cases) and no brain injury group (28 cases).The differences of the composition of gut microbiome between the 2 groups were compared,and the levels of unconjugated bilirubin in serum and cerebrospinal fluid were also compared.Results The level of unconjugated bilirubin in serum of the brain injury group was (463.51 ± 110.62) μmol/L,but in no brain injury group was(364.18 ±63.13) μmol/L,and there was significant difference between the 2 groups(t =4.090,P =0.000 1).The level of unconjugated bilirubin in the cerebrospinal fluid of the brain injury group was (9.53 ± 2.68) μmol/L,but in no brain injury group was (6.94 ± 2.31) μmol/L,and there was significant difference between the 2 groups (t =3.812,P =0.000 3).There was no correlation between the level of unconjugated bilirubin in the cerebrospinal fluid and serum between the 2 groups(r =0.137,0.081,all P ＞0.05).The abundance of gut microbiome in the brain injury group was lower than that in no brain injury group in genus level,among which Fusobacterium,Catabacter,Succinivibrio,Clostridium and Bacteroides were significantly different (all P ＜ 0.05).Conclusions The occurrence of bilirubin brain injury depends on the level of unconjugated bilirubin in serum cerebrospinal fluid,but it may be more directly dependent on the level of bilirubin in the cerebrospinal fluid.The diversity of gut microbiome in neonates with bilirubin brain injury was significantly lower than that in no brain injury group.The level of unconjugated bilirubin in cerebrospinal fluid may be related to the different blood-brain barrier permeability caused by different composition of gut microbiome.

Objective To survey the current situation and influencing factors of willingness to attend assistant general practitioner training among junior medical college students in Yunnan Province.Methods A questionnaire survey and interview on the willingness to attend the training of assistant general practitioner was conducted from May 2017 to June 2017 in Yunnan Province,1 800 students from 6 junior medical colleges were selected as participants by stratified cluster sampling method.Results A total of 1 800 questionnaires were distributed and 1 710 were collected,of which 1 634 were valid with a recovery rate of 95.00％ and an effective rate of 95.56％.The results showed that 43.21％ (706/1 634) participants were not aware of the relevant training policies,while there were significant differences among participants in the different grades and types of college (public or private) (Z=-0.899,8.658,-13.954;P＜0.05);83.17％ (1 379 / 1 634) participants were willing to sign up for an assistant general practitioner training after graduation,while there were significant differences among participants in different ethnic groups,grades,household register sites,types of college,and family income (P＜0.05).Total of 1 300 (79.56％) chose to work as a general practitioner in rural grassroots institutions after graduation,and 984 (60.22％) chose to become a specialist after graduation.Unconditional logistic regression analysis showed that the types of medical college,attitude towards the training policy,the intension to work in grassroots level or to work in private hospitals were influencing factors for willingness to attend the assistant general practitioner training among medical students(B=1.654,3.764,1.361,-0.569;P＜0.05).Conclusion The survey shows that the policy awareness and the willingness to attend assistant general practitioner training for student in junior medical college in Yunnan Province are not high,and the policy publicity and the career guidance need to be strengthened.

Objective To evaluate the survival and prognostic factors of esophageal cancer treated with definitive ( chemo ) radiotherapy by applying novel radiation techniques including three-dimensional conformal radiotherapy (3DCRT) or intensity-modulated radiotherapy (IMRT). Methods Clinical data of 2762 patients with non-operated esophageal squamous cell carcinoma who underwent definitive ( chemo ) radiotherapy from 2002 to 2016 in 10 hospitals were retrospectively analyzed.The prognostic factors were also identified and analyzed. Results The median follow-up time was 60. 8 months. The 1-, 2-, 3-and 5-year overall survival (OS) of all patients was 71. 4％,48. 9％,39. 3％,and 30. 9％,respectively.The 1-,2-,3-and 5-year progression-free survival (PFS) was 59.5％,41.5％,35.2％,and 30％,respectively.The median survival was 23 months.The median time to progression was 17. 2 months.Multivariate analysis demonstrated that age, primary tumor location, clinical stage, tumor target volume, EQD2 and treatment mode were the independent prognostic factors for OS.Primary tumor location,clinical stage,tumor target volume and EQD2 were the independent prognostic factors for PFS. Conclusions In this first large-scale multi-center retrospective analysis of definitive ( chemo) radiotherapy for esophageal squamous cell carcinoma in China, the 5-year OS of patients with esophageal squamous cell carcinoma is significantly improved by 3DCRT, IMRT combined with chemotherapy drugs. However, the findings remain to be validated by prospective clinical trials with high-level medical evidence.

Objective To establish an appropriate neonatal rat model with necrotizing enterocolitis (NEC) and to investigate the protective effects of interleukin-1 receptor associated kinase (IRAK) 1/4 inhibitor on intestinal tissue of neonatal rats with NEC.Methods Thirty-six newborn SD rats were randomly divided into normal control group,NEC model group and IRAK1/4 inhibitor group,12 rats in each group.The rats in normal control group were raised by their mother and they did not receive any intervention.The rats of NEC model group were given artificial feeding,under hypoxia and cold stress.The IRAK1/4 inhibitor group were given IRAK1/4 inhibitor intervention,and also given artificial feeding,under hypoxia and cold stress.Three days later,all rats were sacrificed and intestinal tissues were obtained.The histopathological changes in ileocecal tissues were evaluated by pathological score after hematoxylin-eosin staining.The levels of interleukin (IL)-1β,IL-6 and tumor necrosis factor (TNF)-α in intestinal tissues were detected by enzyme-linked immunosorbent assay.Results The rats of NEC model group presented abdominal distention,diarrhea (partly with mucous bloody stool),decreased activity,poor response,lethargy and other symptoms,and the extent was gradually worsened;the rats in IRAK1/4 inhibitor group also had abdominal distention,diarrhea,decreased activity,poor response and other symptoms,but the symptoms emerged later and milder.The histopathological score of intestinal tissues of normal control group was (0.33 ± 0.49) scores,NEC model group was (3.08 ± 0.99) scores,and IRAK1/4 inhibitor group was (1.75 ±0.96) scores.The histopathological scores of NEC model group and IRAK1/4 inhibitors group were significantly higher than those in normal control group (all P ＜ 0.01),and the histopathological scores of IRAK1/4 inhibitor group were significantly lower than those in the NEC model group (P ＜ 0.01).The levels of IL-1β,IL-6 and TNF-α in normal control group separately were (128.76 ± 27.25) ng/L,(0.41 ± 0.10) ng/L,(6.93 ± 1.79) ng/L,respectively;the levels of NEC model group separately were (410.99 ± 44.16) ng/L,(1.79 ± 0.18) ng/L,(44.39 ± 6.00) ng/L;the levels of IRAK1/4 inhibitor group separately were (256.23 ±41.29) ng/L,(1.05 ±0.19) ng/L,(21.45 ± 6.36) ng/L,respectively.The levels of IL-1β,IL-6 and TNF-α of NEC model group had significant differences compared with those of normal control group,respectively (all P ＜0.01);the levels of IL-1β3,IL-6 and TNF-α of IRAK1/4 inhibitor group had significant differences compared with those of normal control group,respectively (all P ＜ 0.01);the levels of IL-1β,IL-6 and TNF-α of IRAK1/4 inhibitor group had significant differences compared with those of NEC model group (all P ＜ 0.01),respectively.Conclusions IRAK1/4 inhibitor has some protective effects on the intestinal tissues of neonatal rats with NEC,which can reduce the damage to the intestinal tissues,and decrease the secretion of inflammatory cytokines like IL-1β,IL-6 and TNF-α.

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.

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 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 explore the immune mechanism of negative results of immune tests of schistosomiasis japonica pa?tients. Methods Totally 142 schistosomiasis patients(positive stool examinations)of Poyang Lake region were tested by ELI?SA method,and the ROC curve was applied to determine the high and low response of the patients. The levels of cellular immu?nity and cytokines of high and low responders were compared. Results Totally eight schistosomiasis patients were found as low responders. Besides SWAP?IgA(t= -1.588,P > 0.1),the levels of isotype antibodies were significantly lower in the low re?sponders compared with those in the high responders(t = -14.517 to -2.866,all P < 0.05). In the low responders,the propor?tion of CD3+T was increased;and the proportions of CD4+T,CD8+T,CD4+CD25+Treg,and the ratio of CD4+/CD8+ were all de?creased,but all of them were not significant(t = -1.72 to 0.974,all P > 0.05)compared with those in the high responders. The differences of IFN?γ and IL?10 between the high and low responders were both not significant(t= -2.426 to 0.216,all P >0.05). Conclusions There is a significant difference between the high and low responders only in the levels of isotype antibod?ies. One of the reasons of low response in the immune tests is the much lower antibody level after the antigen?antibody compound is completely formulated.