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 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.

Since the oral rehydration salts solution(ORS) has been used to treat acute diarrhea of children, there are different kinds of ORS in order to satisfy the need of clinical work. They are the initial WHO-ORS, other substances-supplemented ORS and the reduced osmolarity ORS, each of which has their own efficiency and advantage.

Objective To investigate the value of the combined detection of serum human epididymal secretory protein 4(HE4) and carbohydrate antigen 125(CA125) in diagnosing ovarian and uterine tumor .Methods Serum levels of HE4 and CA125 were detected in 112 cases of ovarian cancer ,92 cases of uterine cancer ,145 cases of ovarian benign diseases ,138 cases of uterine adeno-myoma and 56 cases of healthy controls by ELISA and electrochemiluminescence(ECL) immunoassay respectively .Results Serum levels of HE4 and CA125 were (246 .80 ± 52 .81)pmol/L and (439 .20 ± 305 .80)U/L in the ovarian malignant tumor group , (196 .50 ± 38 .15)pmol/L and (463 .80 ± 127 .60) U/mL in the uterine malignant tumor group and (19 .76 ± 16 .45)pmol/L and (15 .67 ± 11 .19)U/mL in the control group ,the differences had statistical significance (P<0 .05) .Serum levels of HE4 and CA125 before operation and in postoperative 2 weeks in the ovarian cancer group and the uterine cancer group had statistical difference (P<0 .05) .Conclusion The combination detection of serum HE4 and CA125 can improve the accuracy and sensitivity in the diag-nosis of pelvic tumors .

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Adult ; Coronary Disease ; therapy ; Female ; Humans ; Male ; Middle Aged

Objective To establish a HPLC-ELSD method for determining the content of astragaloside Ⅳ in Qingshen Jianfei Tablets. Methods Stationary phase was C_(18) column (4.6 mm?250 mm, 5 ?m), mobile phase was acetonitrice-water (35 : 65). The evaporation temperature was 120 ℃, the pulverization temperatire was 80 ℃, the flow rate was 1.6 mL/min and column temperature was 35 ℃. Results The standard curve was linear within the range of 1.03～8.24 ?g, r=0.9999. The average recovery was 99.35% with RSD = 1.36% (n = 6). Conclusion The established method is simple and accurate, with good reproducibility and high precision, and suitable for the determination of astragaloside Ⅳ in Qingshen Jianfei Tablets.

Objective:To investigate pathological changes and the expression of the matrix metalloproteinases in rat EAE.Methods:The pathological changes of EAE were studied in Wistar rat with the aid of light and electron microscope and the expression and distribution of MMP-2,MMP-9 in different tissues were detected with the method of immunohistochemistry.Results:Light microscopy showed inflammatory cuff around small blood vessels were evident, disintegration and disappear of myelin sheath were observed. Electron microscopy showed a lot of loose and fragmental spiremes of myelin sheath, the component of axons and nissl bodies in neurons were disappeared. MMP-2,-9 were expressed intensively in vascular endothelial cells, meninges and accumulative inflammatory cells.Conclusion:MMPs take the roles in every aspect of the pathological changes in EAE, it can destroy the blood brain-barrier, degrade the myeline sheath, damage the axons and generate immunogens.

Objective:To investigate establishing the multiple clinical processes model and pathological characteristics of EAE model,and to provide the experimental value for reseaching MS. Methods:The animal model was established in Wistar rat by immunization with complete Freund adjuvant and GPSCH, the pathological changes of EAE were studied with the aid of light microscopy. Results : We can divide the EAE into five types by pathological changes and clinical manifestation: acute EAE, relapsing-remitting EAE, persistent progress EAE,benigh form EAE and delitescence EAE. Every type mainly showed congestion and inflammatory cuff of small blood vessels,disintegration and disappear of myelin sheath and degeneration of neurons,especially in spinal cord. Conclusion:Multiple clinical processes on EAE model of Wistar rat were established for the first time.it is an ideal animal model in studying MS.

Microvesicles (MVs) are the heterogeneous mixtures of vesicles. MVs released by leukemia cells constitute an important part of the leukemia microenvironment. MVs might act as important reservoirs of microRNAs (miRNAs). It is worth evaluating whether MVs possess some unique miRNA contents that are valuable in understanding the pathogenesis. In this study, we investigated the miRNA expression patterns of Nalm-6-derived MVs, Jurkat-derived MVs and normal cell-derived MVs using miRNA microarrays. The potential target genes regulated by differentially expressed miRNAs were also predicted and analyzed. Results demonstrated that 182 miRNAs and 166 miRNAs were differentially expressed in Nalm-6-MVs and Jurkat-MVs, respectively. Many oncogenes, tumor suppressors and signal pathway genes were targeted by these aberrantly expressed miRNAs, which might contribute to the development of B-ALL or T-ALL. Our findings expanded the potential diagnostic markers of ALL and provided useful information for ALL pathogenesis.

The safflower floret is a traditional Chinese medicine used to promote blood circulation and remove obstruction in the channels. The spines on its bracts are considered a handicap when manual harvest is involved. In this study, cDNA-SRAP was used to systematically investigate which genes are associated with the spines. Sixty pairs of possible primer combinations were used on two cDNA pools representing spininess and spinelessness. Six transcript-derived fragments were identified, of which two with low recombination were sequenced successfully and named as GPY-1 and GPY-2. By using the RACE method, the full-length cDNA of GPY-2 is cloned and named as CTL-spn. The full-length cDNA of CTL-spn was 1 679 bp long with a 1 524 bp ORF encoding a 508 aminoacid protein. The deduced amino acid sequence of the CTL-spn gene shared a high homology (97%) with other known ATP synthase CF1 alpha subunits. Semiquantitative RT-PCR analysis revealed that the mRNA of GPY-1 and GPY-2 accumulated in only spiny lines. Considering the important role of ATP synthase CF1 alpha subunit in plants, it may directly take part in the formation process of spininess and enhancing resistance reaction of spiny safflower. Also, our results provide the important insights for breeding spineless cultivars of safflower.
Adenosine Triphosphate ; Amino Acid Sequence ; Carthamus tinctorius ; enzymology ; genetics ; Chloroplast Proton-Translocating ATPases ; genetics ; DNA Primers ; DNA, Complementary ; Plant Proteins ; genetics