BACKGROUND:The hepatic stellate cells(HSCs)plays a key role in the development of liver fibrosis.Studies have shown that bone marrow-derived mesenchymal stem cell(BMSCs)transplantation can be used to treat liver fibrosis,but the mechanism for reversal of liver fibrosis remains unknown.OBJECTIVE:To explore the mechanism of bone marrow mesenchymal stem cells to regulate the proliferation of HSCs under co-culture in vitro.METHODS:Rat BMSCs and HSCs in the experimental group were cultured in the plastic culture plate(6 holes)to establish the upper and lower double-cell co-culture system.Rat normal fibroblast cell lines were seeded as control group;HSCs were cultured alone as blank group.Cell proliferation was determined by WST8 and cell cycle was determined by flow cytometry.The Cyclin D1 and P27 mRNA expression in HSC was determined by reverse transcription-polymerase chain reaction(RT-PCR)and the level of Cyclin D1 and P27 protein by Western blot.RESULTS AND CONCLUSION:HSCs co-cultured with BMSCs significantly inhibited HSC proliferation compared with the blank and control groups at 24,48,and 72 hours(P ＜ 0.01);Flow cytometry showed that the percentage of G_0/G_1 phase cells of co-culture group was increased but the S phase cells reduced(P ＜ 0.01)compared with the other groups at 72 hours,and BMSCs blocked HSC to convert from G_0/G_1 period to S phase.After HSCs co-cultured with BMSCs for 24 hours,the expression of Cyclin D1 mRNA and protein was reduced,and significantly less than the blank and control groups at 72 hours(P ＜ 0.01);no differences were detected in P27 mRNA expression in each group during the co-culture(P ＞ 0.05).After co-culture of 24 hours,the p27 protein expression was significantly increased compared with the blank and control groups(P ＜ 0.01).BMSCs inhibited the proliferation of HSCs,possibly through inhibiting CydinDI expression,increasing the p27 protein expression to cause cell cyde arresting in G_0/G_1 phase.

Objective To investigate the modulatory effects of glycyrrhizin (GL) on the expressions of nu-clear factor kappa B (NF-κB), glucocorticoid receptor (GR) and cytokines in serum, and to explore the protective mechanism of GL in acute lung injury (ALI) induced by lipopolysaecharide (LPS). Method Twenty-four male Sprague-Dawley rats were randomly(random number) divided into three groups (n = 8 in each) : normal group, ALI group and GL treatment group. Rats in the ALI group and GL treatment group were administered with LPS (5 mg/kg) intravenously. In GL treatment group, rats were treated with GL (20 mg/kg) one hour before LPS injec-tion. The animals were sacrificed 4 hours after injection of LPS, and then the lung wewt/dry ratio and PaO_2 were measured, the histopathology of lung injury was observed under light microscope, the expressions of NF-κB mRNA and GR mRNA in lung tissues were detected by using RT-PCR, and the levels of NF-κB protein and GR protein were determined by using Western blot. The levels of TNF-α and IL-10 in serum were observed by using ELISA.Data were analyzed with SPSS version 13.0 software, and means were compared with analysis of variance and Stu-dent-Newman-Keuls test. Results (1) TNF-α levels in normal group, ALI group and GL treatment group were (43.96±7.57), (153.68±20.42), and (87.23±7.52) ng/L, respectively, and IL-10 levels were (24.72±8.03), (42.48±6.81) and (58.33±9.62) ng/L, respectively (F = 183.70, all P <0.01). (2) NF-κB mRNA expressions in normal group, ALI group and GL treatment group were (0.432±0.085), (3.414±0.521) and (1.894±0.272), respectively, and NF-κB protein levels were (45.6±7.3), (254.7±16.4)and (133.5 ±11.7) ng/L, respectively, and comparison between groups showed statistical significant (F = 187.82 and 1466.53, ALL P < 0.01). GR mRNA expressions in normal group, ALI group and GL treatment group were (0.434±0.013), (0.152±0.025) and (0.308±0.033), respectively, and GR protein levels were(54.6±6.5), (11.5±2.3)and (28.2±5.6) ng/L, respectively (F = 246.00 and 260.92, all P < 0.01). (3) Com-pared with normal group, infiltration of PMNs, capillary congestion and swelling were found in ALI group, and treatment with GL could attenuate the lung injury. Conclusions Glycyrrhizin has a protective effects on rats with ALI induced by LPS maybe through down-regulating the expressions of NF-κB mRNA and TNF-α mRNA, and up-regulating the expression of GR mRNA and level of IL-10 protein.

Objective To evaluate measurement of the submucosal thickness with endoscopic ultrasonography (EUS) for activity of Crohn disease (CD).Methods Ten patients with active stage of CD and 10 healthy controls (HC) underwent EUS.Simple endoscopic score for Crohn disease(SES-CD)and submucosal thickness at the most severe lesions were measured and recorded.Submucosal thickness of the same region in CD patients were measured at remissive stage.In order to analyze the relationship between submucosal thickness and the stage of CD, submucosal thickness were compared among patients at active stage of CD, remissive stage of CD and HC.And the cut-off value of submucosal thickness was calculated to diagnose the stage of CD.Results The mean submucosal thicknesses of active stage and remissive stage of CD were 6.48±1.95 mm and 2.47±1.08 mm,respectively (P<0.01).The correlation analysis showed that submucosal thickness had a positive correlation with Crohn disease activity index(CDAI)(r=0.708,P<0.01) and SES-CD(r=0.807,P<0.01).Receiver operating characteristic curve analysis was used for 10 cases of CD patients and the area under the curve was 0.985(P<0.01).The cut-off value of submucosal thickness to diagnose active stage of CD was 3.85 mm, and the sensitivity and specificity reached 100% and 90% respectively.The Youden index was 0.9.Conclusion Measurement of gastrointestinal submucosal thickness by EUS could contribute to evaluate the stage of CD and to guide clinical treatment.

Objective To evaluate the safety and efficacy of endoscopic ultrasonography(EUS) guided ethanol ablation in patients with insulinoma. Methods The data of 10 patients with insulinoma trea-ted at the First Affiliated Hospital of Guangxi Medical University from December 2013 to January 2015 were prospectively analyzed. Results The patients were given EUS-guided ethanol ablation with dose of 0. 10 to 2. 00 ml(average 0. 70 ± 0. 62 ml)in pancreatic lesions for 15 times. No complications were observed dur-ing and after the procedure. The blood glucose improved after the procedure[4. 8(3. 9-5. 5)mmol/ L VS 2. 4 (1. 9-2. 5)mmol/ L,P < 0. 05]and the serum insulin level significantly decreased[83. 7(40. 1-143. 5) pmol/ L VS 177. 3(66. 5-200. 6)pmol/ L,P<0. 05]. The average hospital stay was(4. 3±1. 5)days. The patients were followed up for 6-12 months. EUS indicated that the echo of pancreatic lesions changed from high to low. CE-EUS revealed low enhancement and lack of blood supply. Conclusion EUS-guided ethanol ablation may become a promising minimally invasive treatment for insulinoma because of its safety,efficacy and low price. Trail registration Clinical Trial.gov,NCT02121366.

Objective To study the diagnostic value of narrow-band imaging ( NBI) combined with endoscopic ultrasonography ( EUS) for ampullary tumors. Methods A total of 21 patients suspected with ampullary lesions by imaging or endoscopic examination from December 2015 to March 2017 were enrolled in this prospective study. All patients underwent NBI and EUS, and 20 patients underwent biopsy. The type of ampullary tumor was predicted by preoperative examination, and appropriate treatment methods were chosen. The final diagnosis was confirmed by biopsy, surgical pathology, and clinical follow-up for more than 6 months. The accuracy of NBI combined with EUS and biopsy in diagnosis of ampullary malignant tumors was calculated according to the gold standard. The Chi-square test was used to compare diagnostic accuracies. Results The sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of NBI combined with EUS in diagnosis of ampullary malignancies were 94. 1％ (16/17), 100. 0％ (4/4), 95. 2％ (20/21), 100. 0％ (16/16), and 80. 0％ (4/5), respectively. The corresponding indicators of preoperative biopsy were 41. 2％ ( 7/17) , 100. 0％ ( 3/3) , 50. 0％ ( 10/20) , 100. 0％ ( 7/7) , and 23. 1％( 3/13) , respectively. The accuracy of NBI combined with EUS in diagnosing ampullary malignant tumor was significantly higher compared with preoperative biopsy ( P=0. 004) . Conclusion NBI combined with EUS can more accurately predict benign or malignant ampullary tumor, and better guide the choice of surgical methods compared with preoperative biopsy.