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.

[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 .

Objective To evaluate the curved cutter stapler in the anus-preserving surgery of low rectal cancer. Methods To retrospectively analyse the clinical data of the application of the Curved Cutter Stapler in the anus-preserving surgery of low rectal cancer from 2006 to 2008. Results All the eases of patients with low rectal cancer received the anus-preserving surgery,one of them occurred in sub-clinical small fistula who recovered by con-servative treatment,no anastomofic leakage and no local recurrence occurred,two patients were died in 12,14 months after surgery. Conclusion The Curved Cutter Stapler has an advangtageous position in the anus-preserving surgery of low rectal cancer, can increase the rate of the anus-preserving.

Objective To explore the principle of diagnosis and treatment of early postoperative inflammatory ileus. Method To retrospectively analyse the clinical data and treatment results of early postoperative inflammatory ileus. Results 15 cases after fasting, decompression, nutritional support, somatostatin and dexamethasone, and other conservative treatment were cured. Conclusion Early postoperative inflammatory ileus usually occurred in 2 weeks, has a typical performance of the obstruction, conservative treatment is effective.

Objective:To study the predictive value of the systemic immune inflammation index (SII) on the overall survival rate of patients after Whipple surgery for pancreatic ductal adenocarcinoma (PDAC).Methods:The clinical data of patients with PDAC who underwent Whipple surgery at the First Affiliated Hospital of Zhengzhou University from January 1, 2010 to December 31, 2017 were retrospectively analyzed, and the SII value was calculated. The best cut-off value of SII was 900, and all patients were divided into the low SII group (SII≤900) and the high SII group (SII>900) using 900 as the dividing point. The Kaplan-Meier method was used to draw survival curves and the log-rank test was used. The overall survival of the two groups of patients were analyzed. The Cox risk regression model was used to perform univariate analysis of the various clinicopathological parameters, and multivariate analysis for the statistically significant indicators.Results:Of 135 patients enrolled into this study, there were 78 males and 57 females, aged 28.0-76.0 (56.6±8.8) years. There were 92 patients in the low SII group, aged (56.9±9.2) years with 53 males and 39 females; and 43 patients in the high SII group, aged (56.1±7.9) years, with 25 males and 18 females. The median survival of the low SII group and the high SII group were 32.7 months (95% CI: 28.4-37.0) and 24.4 months (95% CI: 21.4-27.4), respectively. The survival of patients with PDAC in the low SII group was significantly higher than that in the high SII group ( P<0.05). On univariate survival analysis, postoperative overall survival of patients with PDAC was significantly associated with high SII ( HR=2.047, 95% CI: 1.354-3.096), R 1 margin ( HR=2.595, 95% CI: 1.663-4.048), a positive rate of lymph node>20% ( HR=3.244, 95% CI: 1.888-5.573), and positive regional lymph node (N1) ( HR=3.061, 95% CI: 1.993-4.702), all P<0.05. Multivariate Cox regression analysis showed that high SII ( HR=1.672, 95% CI: 1.094-2.555), R 1 resection margin ( HR=2.167, 95% CI: 1.274-3.685), and a positive rate of lymph node>20% ( HR=2.631, 95% CI: 1.309-5.285) to be independent risk factors affecting overall survival ( P<0.05). Conclusion:SII was an independent prognostic factor available before surgery for patients with PDAC. It has some guiding significance in predicting overall survival in patients after Whipple surgery for PDAC.

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.
Animals ; Carbon Tetrachloride/toxicity ; Cells, Cultured ; Hepatic Stellate Cells/physiology ; Liver Cirrhosis, Experimental/*physiopathology ; Male ; Proteins/*physiology ; Random Allocation ; Rats, Wistar ; Transfection ; Wnt Signaling Pathway/*physiology ; beta Catenin/metabolism

Objective To explore the effect of platelet-rich plasma (PRP) on the generation of reactive oxygen species (ROS) and the phenotypes of photo-aging fibroblasts.Methods A photoaging cell model by repeating UVB irradiation was treated using appropriate concentration of PRP;Cell morphology and the rate of aging dying were observed under inverted microscope 24 hours later after establishment of the cell model;The expression of ROS between experimental and control group was detected using fluorescence microscope after single UVB irradiation.The relative intensity of fluorescence was analyzed using flow cytometry.Results PRP could ameliorate the large and sprawl appearance of photoaging fibroblasts obviously,reduce the generation of ROS as well as decrease the relative intensity of ROS.Conclusions PRP can decrease the level of intracellular oxidative stress caused by UVB irradiation,reduce the generation of ROS and ameliorate the senescence-like phenotypes of pho toaging fibroblasts.