Diagnostic criteria for coeliac disease(CD) from the European Society for Paediatric Gastroenterology,Hepatology,and Nutrition were published in 1990.Since then,the autoantigen in CD,tissue transglutaminase had been identified;CD-specific antibody tests had improved.The perception of CD had changed greatly.A panel of 17 experts defined CD and developed new diagnostic criteria based on the Delphi process in 2012.The 2004 National Institutes of Health/Agency for Healthcare Research and Quality report and a systematic literature search on antibody tests for CD in paediatric patients covering the years 2004 to 2009 was the basis for the evidence-based recommendations on CDspecific antibody testing.In group of children with symptoms suggestive of CD,the diagnosis of CD is based on symptoms,positive serology,and histology that is consistent with CD.If immunoglobulin A anti-tissue transglutaminase type 2 antibody titers are high (＞ 10 times the upper limit of normal),then the option is to diagnose CD without duodenal biopsies by applying a strict protocol with further laboratory tests.In group of asymptomatic children at increased risk for CD,the diagnosis of CD is based on positive serology and histology.Human leukocyte antigen-DQ2 and human leukocyte antigen-DQ8 testing is valuable because CD is unlikely if both haplotypes are negative.The aim of the new guidelines was to achieve a high diagnostic accuracy and to reduce the burden for patients and their families.

Objective To observe the clinical effects of short-term intermittent ganeiclovir treatment on infants with cytomegalovirus (CMV) hepatitis. Methods The clinical data of infants with CMV hepatitis were analyzed retrospectively. The liver functions including total bilirubin (TBil), direct bilirubin (DBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and γ-glutamyltransferase (γ-GT) of the patients in treatment group (85 cases) and control group (37 cases) were collected before and after treatment. Meanwhile, the side effects of ganciclovir during treatment were observed. The measurement data were compared by analysis of variance and numeration data were compared by chi-square test. Results After short-term intermittent ganciclovir treatment in treatment group, TBil level was decreased from (109.1±677.8)μmol/L to (62.9±68.1)μmol/L (F=15.34,P<0.01); ALT level was decreased from (160.2±395.3) U/L to (68.1±56.0) U/L (F=4.73, P<0.05). In control group, TBil level was decreased from (94.9±647.4)μmol/L to (49.2±631.5) μmol/L (F=14.80, P<0.01) ; while ALT level was decreased from (131.6±206.2) U/L to (55.3±631.2) U/L (F=3.50, P=0.067). The readmission rate in control group was significantly higher than that in treatment group (21.6% vs 10.6%). Only one case (0.8%) received three times of intermittent ganciclovir treatment. The longest hospitalization time was six weeks. Conclusions Short-term intermittent ganciclovir treatment may be more suitable for infants with CMV hepatitis. There is no obvious side effect observed during the treatment and the hospitalization time can be shortened.

Objective To carry out the detection of JAGI gene in children with chronic cholestasis and to im-prove the diagnostic level and understanding of Alagille syndrome. Methods Two cases of chronic cholestasis with multiple organ involvement were selected as the research subjects and their clinical data,laboratory test results were col-lected. Two milliliter peripheral intravenous heparin anticoagulan blood was drawn from each patient. All fragments of 26 exons of the JAGI gene were amplified by polymerase chain reaction - sequence based on typing method. Results One patient with chronic cholestasis,heart murmur and dysmorphic face showed bile duct paucity in liver biopsy and a novel heterozygous mutation c. 809 809delG(p. G270Dfs*142)in 6 exon. Abnormal amino acid replaced JAG1 protein and resulted in truncation of the JAG1 protein. The part of epidermal growth factor(EGF)like repeats region loss and the cysteine rich region completely lost. One case with typical chronic cholestasis and dysmorphic face showed a known IVS20 - 2 5delTAAG heterozygous mutation which resulted in splice site changes. Conclusion A novel JAGI gene mutation c. 809 809delG(p. G270Dfs*142)is helpful to screen JAGI gene of Notch signal transduction pathway for chronic cholestasis with multiple organs involvement in children.

Objective To research the effects of different titers of bacteriophage D29 on growth and function of airway epithelial cell 9HTE.Methods Cell viability rates was analyzed after applying high(109 PFU/mL)and low(107 PFU/mL)titers of bacteriophage D29 and phage buffer respectively by MTT colorimetry.Additionally,the secretion levels of IL-6,IL-8 in cell culture supernatant were detected by ELISA.RT-PCR was performed to detect the expression of ICAM-1 mRNA.Cell apoptosis rate was analyzed by flow cytometry.Results There was no difference in cell growth,secretion levels of IL-6,IL-8,ICAM-1 mRNA and cell apoptosis rate between cells treated with high and low titers of D29 and phage buffer(P>0.05).Conclusion Neither high nor low titer of bacteriophage D29 exerts effect on growth and function of airway epithelial cell 9HTE in vitro.

OBJECTIVE:To study the preparation technology of Fanhuncao sustained-release dropping pills. METHODS:The preparation technology of Fanhuncao sustained-release dropping pills was optimized by orthogonal design and single factor test with PEG4000,PEG6000 and glycerol monostearate(GM)as carrier materials,using sphericity,pill weight difference and appearance quality as index,GM-PEG weight ratio,liquid temperature,nozzle temperature,dipping speed,dropping distance,diameter of emitter as factors and the verification test was conducted. RESULTS:The optimal technology was that the ratio of GM-PEG was 3∶7;the temperature of drug mixture was 85 ℃;the dropping speed was 40 drop/min;dropping distance was 4 cm;and the conden-sate temperature was 15 ℃;diameter of emitter was 4 mm. RSD of each index of 3 batches of dripping pills were all ≤0.58%(n=3)in verification test;the average content of chlorogenic acid was 0.14 mg/g;the average pill weight difference was 3.21%;the average sphericity was 9.43;and the average appearance quality was 4.33. Q1 h were 23.4%,24.4% and 23.3% in average(n=6),and Q12 h were 89.6%,91.2% and 91.5%(n=6). CONCLUSIONS:The optimal preparation technology is stable and simple, and can be used for industrial production of Fanhuncao sustained-release dropping pills.

Objective To investigate the incidence of neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD)in neonates with idiopathic neonatal cholestasis (INC)in Nanjing,China,SLC25A13 gene mutations in these neonates,and clinical features.Methods A total of 152 neonates with INC,who were admitted to the Affiliated Nanjing Children's Hospital of Nanjing Medical University from Sep-tember 2009 to August 2013,underwent gene analysis for detecting SLC25A13 gene mutations.The neonates were divided into NICCD group,who had been diagnosed definitely,and INC group at a ratio of 1∶2,considering the age and gender.Several biochemical indices were compared between the two groups.Comparison of continuous data between the two groups was made by Mann-Whitney U test after Bonferroni correction.Results There were 21 confirmed cases of NICCD (21/152,13.82%)among the 152 neonates with INC;five types of SLC25A13 mutations were identified in the 21 neonates with NICCD,including 851_854del (27/42,64.29%),IVS6+5 G→A (7/42, 16.67%),1638ins23 (5/42,11.90%),IVS11 +1 G→A (2/42,4.76%),and Q259X (1/42,2.38%).The alanine aminotransferase (ALT)level,aspartate aminotransferase (AST)level,bile acid concentration,albumin level,fasting blood glucose,blood ammonia,and prothrombin time for the NICCD group were 39.42 ±23.40 U/L,124.85 ±92.65 U/L,142.43 ±24.34μmol/L,30.66 ±2.70 g/L,2.79 ± 0.54 mmol/L,117.57 ±27.88 μmol/L,and 14.03 ±2.79 s,respectively,versus 136.02 ±113.67 U/L,226.12 ±129.26 U/L,80.47 ± 31.53 μmol/L,36.87 ±4.96 g/L,3.14 ±0.45 mmol/L,76.43 ±20.80 μmol/L,and 11.40 ±1.55 s for the INC group.The NICCD group had significantly lower ALT and AST levels than the INC group (Z=-5.02,P=0.000;Z=-3.66,P=0.000);the NICCD group had a significantly higher bile acid concentration than the INC group (Z=-5.58,P=0.000);the NICCD group had significantly lower albumin level and fasting blood glucose than the INC group (Z=-4.52,P=0.000;Z=-2.56,P=0.010);the NICCD group had a significantly higher blood ammonia level than the INC group (Z=-4.75,P=0.000);the NICCD group had a significantly longer prothrombin time than the INC group (Z=-4.10,P=0.000).Conclusion Citrin deficiency due to SLC25A13 gene mutations is an im-portant cause of INC in Nanjing.The three most common mutations are 851_854del,IVS6+5 G>A,and 1638_1660dup23,which account for 92.86% of the SLC25A13 gene mutations.More attention should be paid to clinical analysis and detection of SLC25A13 gene mutations to confirm the diagnosis of NICCD.

Cytochrome P450 (CYP450) is a key element in the Ganoderma triterpenoid biosynthetic pathway. The catalytic reaction process for CYP450 requires NADPH / NADH for electron transfer. After searching the genome dataset of Ganoderma lucidum, the unique sequence encoding CYP450 and NADPH were discovered, separately. The open reading frames of GLCYP450 and GLNADPH were cloned separately using RT-PCR strategy from G lucidum. The appropriate restriction enzyme cutting sites were introduced at the 5' and 3' ends of gene sequence. The genes of GLCYP450 and GLNADPH were recombined into the yeast expression vector pESC-URA, leading to the formation of the yeast expression plasmid pESC-GLNADPH-GLCYP450. This study provides a foundation for researching Ganoderma triterpene biosynthesis using the approach of synthetic biology.

ObjectiveTo analyze the incidence,etiology and clinical treatment of the impacted maxillary central incisors.Methods A retrospective study was conducted on 29 cases of impacted maxillary central incisors that were randomly selected from the 1000 patients.Age of the patient was ranged from 7 to 23 years with median 11.1 years.Numbers of impacted teeth were 32,in which 22teeth were intraosseous impaction and others were extraosseous.The incidence,etiology,sites and direction of impacted teeth and their treatment approaches were explored in this study.Results Impac tion of maxillary central incisors occurred more frequently in females than in males,with a ratio of 1.0 ∶ 1.2.The ratio of intraosseous impaction to extraosseous one was 11 ∶ 5.The insufficient eruption space of maxillary central incisors was present in 65％ patients,but intermediate and severe crowded teeth only accounted for 15％.The incidence of the abnormal site and position of impacted teeth reached 71 ％.The clinical approach and solution were surgical-orthodontic treatment,transplantation or removal of impacted teeth.ConclusionsThe main cause is abnormity of teeth in the shape and position.Most of the impacted maxillary central incisors could be moved to the correct position by effective treatment.

e genotypes.