Clinical significance of serum Pygo2 expression in children with NAFLD / 医学研究生学报

[Abstract ] Objective The role of serum Pygo2 expression in children with non-alcoholic fatty liver disease(NAFLD) in the disease process and whether it can be used to evaluate the degree of liver fibrosis still remain unknown .The article aimed to detect Pygo 2 expression in the peripheral blood of children with NAFLD and ana-lyze its relationship with traditional serum hepatic fibrosis index in or-der to evaluate the clinical significance of Pygo 2 measurement in chil-dren with NAFLD . Methods We enrolled 120 cases of childhood obesity and 18 healthy controls in Anhui Provincial Hospital and the First Affiliated Hospital of Nanjing Medical University from September 2014 to February 2016.The cases of childhood obesity were di-vided into simple obesity group ( n=44, no diffuse fatty liver under liver ultrasound detection ) , NAFL group ( n=35, diffuse fatty liver with normal liver function under liver ultrasound detection ) and NASH group ( n=41, diffuse fatty liver with abnormal liver function under liver ultrasound detection ) .The peripheral serum was collected from all patients and healthy controls .ELISA was used to detect serum Pygo2 expression and radioimmunoassay was used to detect the serum hyaluronia acid (HA), procollagen type Ⅲ(PCⅢ), pro-collagen type Ⅳ(CⅣ) and laminin(LN) levels.Finally the serum ALT and γ-GT levels were measured with totally automatic enzymat-ic method. Results Pygo2 expression in NAFL group [52.1(12.3)μg/L] and NASH group[78.3(50.0)μg/L] increased signifi-cantly compared with simple obesity group [43.2(18.7)μg/L](P<0.05).Pygo2 expression in NASH group increased significantly compared with control group [41.7(16.8)μg/L] and NAFL group (P<0.001).The serum hepatic fribrosis and inflammation markers ( HA, PC, ALT, C III and IV gamma-GT) levels gradually increased in the obese children .There were statistically positive correla-tions between serum Pygo2 and HA, PCⅢ, CⅣ, ALT or γ-GT (P<0.001).In particular, more significant positive correlations were found between serum Pygo2 and HA, CⅣor γ-GT (r=0.708, P<0.001;r=0.589, P<0.001;r=0.674, P<0.05). Conclusion The degree of liver fibrosis worsens with the increase of Pygo 2 expression and Pygo 2 is in remarkable correlation with conventional he-patic fibrosis serum makers ( HA, C IV) orγ-GT, which shows Pygo 2 expression can be taken as the clinical evaluation index of liver fibrosis in children with NAFLD .

Mest Attenuates CCl4-Induced Liver Fibrosis in Rats by Inhibiting the Wnt/beta-Catenin Signaling Pathway

BACKGROUND/AIMS: The Wnt/beta-catenin signaling pathway has been reported to play an important role in liver fibrosis. This study was designed to investigate whether mesoderm-specific transcript homologue (Mest), a strong negative regulator of Wnt/beta-catenin signaling, could inhibit liver fibrosis. METHODS: pcDNA-Mest was transfected into hepatic stellate cells (HSCs) and rats. Rats were randomly divided into four groups: normal group (normal saline), treatment group (pcDNA-Mest+CCl4), control group (pcDNA-neo+CCl4), and model group (normal saline+CCl4). Changes in liver pathology were evaluated by hematoxylin and eosin and Masson's trichrome staining. The levels of alanine transaminase, aspartate transaminase, lactic dehygrogenase, hyaluronic acid, and laminin in the serum and hydroxyproline in the liver were detected by biochemical examination and radioimmunoassay, respectively. The expression and distribution of beta-catenin, alpha-smooth muscle actin (alpha-SMA), Smad3, and tissue inhibitor of metalloproteinase type I were determined, and the viability of the HSCs was tested. RESULTS: Our data demonstrate that Mest alleviated CCl4-induced collagen deposition in liver tissue and improved the condition of the liver in rats. Mest also significantly reduced the expression and distribution of beta-catenin, alpha-SMA and Smad3 both in vivo and in vitro, in addition to the viability of HSCs in vitro. CONCLUSIONS: We found that Mest attenuates liver fibrosis by repressing beta-catenin expression, which provides a new therapeutic approach for treating liver fibrosis.

Construction and characterization of hfgl 2 expression vector / 医学研究生学报

Objective:To construct hfgl 2 expression vector(pcDNA3.1-hfgl 2) and characterize the expression of hfgl 2 in CHO cells after transfection. Methods: Total RNA was extracted from chinese human peripheral blood monocyte cells,cDNA was obtained by reverse transcription,hfgl 2 cDNA was amplified and cloned into pcDNA3.1 and the orientation and the sequence were ensured by restriction endonucleases and sequencing assays.The recombinated plasmid was transfected into CHO cells,and the expression of hfgl 2 was detected by immunohistochemistry. Results:A 1.3 kb long target fragment was obtained and cloned into pcDNA3.1.The orientation and sequence are correct.hfgl 2 was only expressed in those cells transfected with pcDNA3.1-hfgl 2. Conclusion:hfgl 2 expression vector(pcDNA3.1-hfgl 2) has been successfully constructed.