1.The expression and clinical significance of matrix metalloproteinase-2 in human pancreatic carcinoma.
Xuefeng ZHU ; Yijun CHEN ; Jili GU ; Dechun LI ; Dongming ZHU
Clinical Medicine of China 2009;25(10):1032-1034
Objective To investigate the expression of matrix metalloproteinase-2 (MMP-2)in pancreatic carcinoma and the relationship between MMP-2 with tumour clinicopatholngical features. Methods The expression of MMP-2 was detected by S-P immunohistochemistry in 36 cases with pancreatic carcinoma and 14 normal pancreat-ic tissues. Results The positive expression rate of MMP-2 was 66.7% (24/36) in pancreatic carcinoma tissue and 14.3% (2/14) in normal pancreatic tissues (χ2 = 3. 587, P < 0.01 ) ;The expression rate of MMP-2 in pancreatic carcinoma tissue with positive-node was 86.7% ( 13/15 ) ,which was higher than that with negative-node,which was 52.4% ( 11/21 ) ( P < 0.05 ) ; As to TNM staging in pancreatic carcinoma, the expression rate of MMP-2 was 41.2% (7/17) with Ⅰ,Ⅱ staging and 89.5% (17/19) with Ⅲ,Ⅳ staging(χ2=9.418,P <0. 01 ) ;The expression rate of MMP-2 was 50.0% (5/10) ,66.7% (10/15) and 72.8% (8/11) in highly,moderately and poorly differentiated pancreatic carcinoma(P > 0.05 ). Conclusions The expression of MMP-2 is strengthened significantly in pancreatic carcinoma tissue and involved in turnout invasion and metastasis features; MMP-2 might be regarded as one more marker for the invasive properties of pancreatic carcinoma.
2.Inhibition of invasiveness of pancreatic carcinoma cell line PANC-1 by suppression of MMP-2 gene expression using RNA interference
Xuefeng ZHU ; Dechun LI ; Yijun CHEN ; Jianwei XU ; Jili GU ; Dongming ZHU ; Jiannong CEN
Chinese Journal of Hepatobiliary Surgery 2010;16(11):863-866
Objective To investigate the inhibitory effects of RNA interference on expression of matrix metalloproteinase-2(MMP-2)gene and invasiveness of human pancreatic cancer cell line PANC-1 in vitro.Methods Small interference RNA targeting MMP-2 gene was designed and constructed to plasmid pGCsi-U6.Recombinant plasmids were transfected to pancreatic carcinoma PANC1 cells with Lipofectamine 2000.The efficiency of transfection was evaluated by flow cytometry.RQPCR was used to detect the expression of MMP-2 mRNA.The expression of MMP-2 protein was determined by ELISA.The invasiveness of PANC-1 cells was measured by transwell chamber experiment.MTT assay was used to detect the proliferation and growth of PANC-1 cells.Results Sequencing confirmed that the MMP-2 siRNA plasmid was successfully constructed.The best efficiency of transfecting recombinant plasmid was 82.1%.After transfection of the MMP-2 siRNA plasmid, the MMP-2 gene expression of PANC-1 cells was suppressed to 71.74 %(P<0.05),and protein expression of MMP-2 fell to 49.82%(P<0.05).The corresponding inhibition ratio of invasiveness was 33.0%(P<0.05).There was no marked difference in proliferation rate measured by MTT assay among different groups(P>0.05).Conclusions RNAi targeting MMP-2 can suppress invasiveness of PANC-1 cells in vitro.This suggests that MMP-2 could be a target for gene therapy of pancreatic carcinoma.RNAi is expected to open up a new prospect for tumor therapy.
3.Comparative analysis of intestinal microbiota distribution characteristics based on metagenomics in patients with hepatitis B cirrhosis with or without ascites
Yuting LUAN ; Chenghai LIU ; Shili JIANG ; Hongtu GU ; Jing LYU ; Feng XING ; Changqing ZHAO ; Jili YUAN ; Ping LIU ; Yongping MU
Chinese Journal of Hepatology 2023;31(9):974-985
Objective:To use metagenomic sequencing to compare the differences in intestinal microbiota species and metabolic pathways in patients with hepatitis B cirrhosis with or without ascites and further explore the correlation between the differential microbiota and clinical indicators and metabolic pathways.Methods:20 hepatitis B cirrhosis cases [10 without ascites (HBLC-WOA), 10 with ascites (HBLC-WA), and 5 healthy controls (HC)] were selected from the previously studied 16S rRNA samples. Metagenome sequencing was performed on the intestinal microbiota samples. The Kruskal-Wallis rank sum test and Spearman test were used to identify and analyse differential intestinal microbiota populations, metabolic pathways, and their correlations.Results:(1) The overall structure of the intestinal microbiota differed significantly among the three groups ( R = 0.19, P = 0.018). The HC group had the largest abundance of Firmicutes and the lowest abundance of Proteobacteria at the genus level. Firmicutes abundance was significantly decreased ( Pfdr < 0.01), while Proteobacteria abundance was significantly increased ( Pfdr < 0.01) in patients with cirrhosis accompanied by ascites; (2) LEfSe analysis revealed that 29 intestinal microbiota (18 in the HBLC-WA group and 11 in the HBLC-WOA group) played a significant role in the disease group. The unclassified Enterobacteriaceae and Klebsiella species in the HBLC-WA group and Enterobacteriaceae in the HBLC-WOA group were positively correlated with the Child-Turcotte-Pugh (CTP) score, prothrombin time, and international normalized ratio score and negatively correlated with albumin and hemoglobin levels ( P < 0.05). Escherichia and Shigella in the HBLC-WA group were positively correlated with CTP scores ( P < 0.05); (3) The correlation analysis results between the KEGG pathway and 29 specific intestinal microbiota revealed that Enterobacteriaceae and arachidonic acid, α-linolenic acid, glycerolipid metabolism, and fatty acid degradation were positively correlated in the lipid metabolism pathway, while most Enterobacteriaceae were positively correlated with branched-chain amino acid degradation and negatively correlated with aromatic amino acid biosynthesis in the amino acid metabolic pathway. Conclusion:A significant increment of Enterobacteriaceae in the intestines of HBLC-WA patients influenced hepatic reserve function and was associated with amino acid and lipid metabolic pathways. Therefore, attention should be paid to controlling the intestinal microbiota to prevent complications and improve the prognosis in patients with hepatitis B cirrhosis, especially in those with ascites.
4.Effect of traditional Chinese medicine on portal vein thrombosis in patients with liver cirrhosis and its medication characteristics
Yanan GUO ; Hongtu GU ; Changqing ZHAO ; Yongping MU ; Jili YUAN ; Feng XING ; Yanyan TAO ; Jing LYU ; Chenghai LIU
Journal of Clinical Hepatology 2023;39(2):345-351
Objective To investigate the therapeutic effect of traditional Chinese medicine (TCM) on portal vein thrombosis (PVT) in patients with liver cirrhosis and its medication characteristics. Methods A retrospective analysis was performed for 89 patients with liver cirrhosis and PVT who were hospitalized and treated in Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, and according to whether TCM treatment was applied in combination, they were divided into TCM group with 59 patients and control group with 30 patients. Related data were collected for the two groups, including demographic data, laboratory examination, radiological examination, gastroscopy, history of surgery, portal hypertension-related complications, medication, and follow-up data. The independent samples t -test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U rank sum test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test or the Fisher's exact test was used for comparison of categorical data between two groups. An ordinal polytomous Logistic regression analysis was used for multivariate analysis. TCM Inheritance Computing Platform (V3.0) was used to perform a drug effect cluster analysis of TCM prescriptions. Results The multivariate logistic regression analysis showed that esophageal and gastric varices (odds ratio [ OR ]=3.144, 95% confidence interval [ CI ]: 1.221-8.094), PVT involving the portal vein (PV) and the superior mesenteric vein (SMV) ( OR =51.667, 95% CI : 3.536-754.859), PVT involving PV+spleen vein (SV)+SMV ( OR =13.271, 95% CI : 2.290-76.928), cavernous transformation of the portal vein ( OR =11.896, 95% CI : 1.172-120.696), and TCM intervention ( OR =0.348, 95% CI : 0.129-0.938) were influencing factors for the outcome of PVT in liver cirrhosis. Follow-up results showed that compared with the control group, the TCM group had a significantly lower progression rate (16.95% vs 56.67%, P < 0.001) and a significantly lower incidence rate of variceal rupture and bleeding (8.47% vs 33.33%, P < 0.001). Effective TCM drugs with a relatively high frequency of use included deficiency-tonifying drugs (359 times, 34.6%), blood-activating and stasis-resolving drugs (202 times, 19.5%), and diuresis-inducing and dampness-draining drugs (180 times, 17.3%); the TCM drugs with a relatively high frequency of use included Astragalus membranaceus (57 times, 8.7%), Angelica sinensis (50 times, 7.6%), and leech (48 times, 7.3%); TCM drug combinations with a relatively high frequency of use included Astragalus membranaceus+Angelica sinensis, Astragalus membranaceus+leech, Angelica sinensis+leech, and Astragalus membranaceus+Angelica sinensis+leech. Conclusion Qi-tonifying, blood-activating, and stasis-breaking drugs, such as Astragalus membranaceus, Angelica sinensis, and leech, can promote the stabilization or recanalization of PVT in liver cirrhosis and reduce the incidence rate of bleeding events due to portal hypertension.