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Liver cirrhosis, characterized by diffuse hepatocytes necrosis, insufficient regeneration of hepatocytes, angiogenesis, severe fibrosis and the formation of pseudolobules, is a progressive chronic hepatic disease induced by a variety of causes. It is clinically characterized by liver function damage and portal hypertension, and many complications may occur in the late stage. Based on the update relevant guidelines, experts' consensus, and research advances on the diagnosis and treatment of cirrhosis, Chinese Society of Gastroenterology of Chinese Medical Association established a consensus aiming to standardize the clinical diagnosis and treatment of liver cirrhosis and guide clinical practice. This consensus contains 43 statements on the etiology, pathology and pathogenesis, clinical manifestations, major complications, diagnosis, treatment, prognosis and chronic disease management of liver cirrhosis. Since several guidelines and experts' consensus on the complications of liver cirrhosis have been published, this consensus focuses on the research progress of liver cirrhosis itself.
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Objective@#To evaluate Chinese clinical practice guideline/consensus for digestive diseases published in the past five years in order to recommend the high-quality guidelines and help with the promotion and implementation of them.@*Methods@#From January 2013 to June 2018, the officially published Chinese practice guideline/consensus for digestive diseases were selected. The inclusion and exclusion criteria of the guideline/consensus was evaluated by "Evaluation Criteria for Chinese Clinical Practice Guidelines 2017(AGREE-China 2017)" . The guideline/consensus were independently scored by three evaluators and then calculated the average value. Descriptive analysis methods were used to analyze the Chinese clinical practice guideline/consensus for digestive diseases. Those with the total score more than 40.0 points were included in the recommended list.@*Results@#A total of 119 officially published clinical practice guideline/consensus of digestive diseases were retrieved, and 74 clinical practice guideline/consensus for digestive diseases were included in the evaluation. Among them, 18 (24.3%, 18/74) scored over 60.0 points, 31 (41.9%, 31/74) scored between 40.0 and 59.9 points. Finally 48 guideline or consesus were selected for the recommended list 19 cases of esophagus and gastrointestinal diseases, 18 cases of liver diseases, five cases of biliary and pancreafic diseases, and six cases of digestive endoscopy. The three guideline/consensus with the high scores (> 80.0 points) were The Fifth Chinese National Consensus Report on the Maragement of Helicobacter pylori Infection, Consensus on the Diagnosis and Treatmeat of Cholestatic Liver Disease (2015) and Guidelines for the Prevention and Treatment of Chroaic Hepatitis B (2015 Update). The higher the score of the guideline/consensus, the more scientific and rigorous the method, and the clearer the evaluation of evidence grade and the description of the formation of recommendations. Compared with international standards of guideline/consensus development, there are still some problems in Chinese guidelines or consensus such as no explanation of retrieval strategy, no basis of evidence classification and no description of the formation process from evidence to recommendation.@*Conclusions@#The quality of Chinese clinical practice guideline/consensus for digestive diseases has been improved year by year. However the scientific aspects need to be further improved. AGREE-China which demonstrates good validity, realiability and practicability is easy and clear to use.
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Objective To evaluate Chinese clinical practice guideline/consensus for digestive diseases published in the past five years in order to recommend the high-quality guidelines and help with the promotion and implementation of them.Methods From January 2013 to June 2018,the officially published Chinese practice guideline/consensus for digestive diseases were selected.The inclusion and exclusion criteria of the guideline/consensus was evaluated by " Evaluation Criteria for Chinese Clinical Practice Guidelines 2017 (AGREE-China 2017)".The guideline/consensus were independently scored by three evaluators and then calculated the average value.Descriptive analysis methods were used to analyze the Chinese clinical practice guideline/consensus for digestive diseases.Those with the total score more than 40.0 points were included in the recommended list.Results A total of 119 officially published clinical practice guideline/consensus of digestive diseases were retrieved,and 74 clinical practice guideline/consensus for digestive diseases were included in the evaluation.Among them,18 (24.3％,18/74) scored over 60.0 points,31 (41.9％,31/74) scored between 40.0 and 59.9 points.Finally 48 guideline or consesus were selected for the recommended list 19 cases of esophagus and gastrointestinal diseases,18 cases of liver diseases,five cases of biliary and pancreafic diseases,and six cases of digestive endoscopy.The three guideline/consensus with the high scores (＞ 80.0 points) were The Fifth Chinese National Consensus Report on the Maragement of Helicobacter pylori Infection,Consensus on the Diagnosis and Treatmeat of Cholestatic Liver Disease (2015) and Guidelines for the Prevention and Treatment of Chroaic Hepatitis B (2015 Update).The higher the score of the guideline/consensus,the more scientific and rigorous the method,and the clearer the evaluation of evidence grade and the description of the formation of recommendations.Compared with international standards of guideline/consensus development,there are still some problems in Chinese guidelines or consensus such as no explanation of retrieval strategy,no basis of evidence classification and no description of the formation process from evidence to recommendation.Conclusions The quality of Chinese clinical practice guideline/consensus for digestive diseases has been improved year by year.However the scientific aspects need to be further improved.AGREE-China which demonstrates good validity,realiability and practicability is easy and clear to use.
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Objective To analyze the clinical and histological features of drug-induced autoimmune hepatitis (DI-AIH).Methods From January 2008 to June 2011,five patients with auto-immune hepatitis (AIH),having a definitive history of medicine taking prior to the onset of disease and accepted liver biopsy were retrospectively analyzed.The general information was collected,which included gender,age,onset of the disease,medication,clinical manifestations,treatment and follow-up.The laboratory findings were also collected,which included total bilirubin (TBil),direct bilirubin (DBil),alanine transaminase (ALT),aspartate transaminase (AST),alkaline phosphatase (ALP),gamma glutamyl transpeptidase (GGT),globulin,γ-globulin percentage,albumin (Alb),eosinophils,antinuclear antibodies,antimitochondrial antibodies,anti-smooth muscle antibodies and the type of live injury.The liver tissue sections of patients were stained with hematoxylin-eosinstaining (HE) and reticular fiber staining and then pathological changes were observed.Patients with DI-AIH were scored with AIH scoring scale.Results All five patients with DI-AIH were female,average age was 48.0--+7.5.Prior to the onset of disease,all patients had taken Chinese traditional medicine.Anorexia and fatigue were the most common clinical symptoms.Among laboratory findings,the level of ALT ((795.0+467.4) U/L),AST ((730.44-451.5) U/L),TBil ((80.3-+ 64.1) μmol/L) and DBil ((65.2 +_ 58.0) μmol/L) significantly increased.One patient was antinuclear antibody positive.One patient had drug-induced liver injury,pathological features were spotty necrosis of liver cells,liver tissue eosinophil infiltration and liver cell microbubble type degeneration.All the patients had interface hepatitis,periportal infiltration of lymphocytes or lymphocytes-plasma cells,liver cells adjacent to lesion showed rosette-like structure.All the patients received glucocorticoid treatment.After glucocorticoid withdrawal,the liver function was normal during the follow-up period.Conclusion There are no specificity of clinical manifestations,laboratory findings and histological features in patients with DI-AIH.
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Objective To investigate the application effect of Problem based learning (PBL) combined with role play teaching method in the training of emergency internal medicine.Methods Totally 60 medical students were randomly divided into two groups.The students in the observation groop were trained with PBL teaching method while those in control group were trained with traditional teaching method.The practical performances of the students in both groups were evaluated by virtual experience of dealing with patients in the emergency department through the role play teaching method.The grasp of basic knowledge was assessed by writing tests.Finally all the students in the observation group answered a survey about the teaching methods.Results There was uo difference in the scores of the basic knowledge test between the two groups ( P ＞ 0.05 ).The practical ability of the observation group was much better than that of the control group in the role play practice ( P ＜ 0.05 ).Almost all the students in the observation group accept the PBL teaching method.Conelusion PBL combined with role play teaching method is effective based on the characteristics of emergency medicine.
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Objective To comprehensively evaluate the clinical and radiological features of patients with Klebsiella pneumonia liver abscess (KLA) in order to improve treatment. Methods Data of 197 patients with KLA (n=106) or non-KLA (n=56) between March 2001 and January 2009 were collected and retrospectively analyzed on the aspects of underlying diseases, clinical manifestations, laboratory tests, pathogenic and imaging examinations. Results In comparison with non-KLA patients, the underlying diseases such as diabetes (53. 77% vs 25.00%, P = 0. 001) and hepatic adipose infiltration (16. 04% vs 5, 36%, P<0.05) were more common in KLA patients.Whereas the clinical presentations including abdominal pain (57. 14% vs 40. 57%, P<0. 05),hypodynamia (46.43% vs 19.81%, P=0. 001) and hepatomegaly (14.29% vs 4.72%, P<0. 05)were more severe in non-KLA patients than in KLA patients, however, the fasting blood glucose was higher in KLA patients than in non-KLA patients [(7.84±0.36) mmol/L vs (5.76±0.30) mmol/L,P=0. 001] on the admission. In addition, the abscess of KLA often appeared alone in the right lobe of the liver and was liable to generate air cavity (32. 88% vs 13.51%, P<0.05), un-smooth rim (71.23% vs 40.54% ,P<0.05) and dynamic septum enhancement (41.10% vs 16.22% ,P<0.01) in comparison with non-KLA. Conclusions Klebsiella pneumonia has emerged as the main pathogen of pyogenic liver abscess. The patients with KLA are often complicated with diabetes and fatty liver, as well as high prevalence of air cavity. The CT findings may be helpful for prompt treatment of Klebsiella pneumonia liver abscess.
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Objective To analyse the clinical characteristics,follow up their prognsis and evaluate the treatment of patients with Crohn' s disease (CD). Methods The data of patients with CD were collected at Zhongshan Hospital of Fudan University from 2002 to 2007. The diagnosis was made according to the consensus recommended by Chinese Society of Gastroenterology. The disease severity, follow-up and prognsis were evaluated according to the consensus. Results Sixty-six CD patients were enrolled in this study. The ratio of males to females was 2.47 : 1. Age at diagnosis ranged from 12 to 76 years old, and the mean age was (32 ± 17)years old. The main gastroenterological manifestations were abdominal pain (80.3 %), diarrhea (54.6 %), fistula formation (31.8%) ; and the main systemic manifestation was fever (80.3 %) and defective nutrition. A total of 60.6 % of patients with CD had complications and 34. 8% of these patients had more than one complications. Extraintestial manifastation were presented in 7.6% of patients with CD. A total of 86.4% of patients with CD were active state cases and 57. 6% were suffered from inflammation of ileum and colon. Abdominal pain (P = 0. 011), hematochezia (P = 0. 008), fever (P = 0. 001), anemia (P = 0. 020), underweight (P = 0. 010) and heightening of CRP (P = 0. 033) were more common in patients who were in active state than those who were in slient state. During their inducing alleviate treatment, 33.3% of patients with CD were treated with aminosalicylate alone, 36.4% were treated with both aminosalicylate and glucocorticoids, 7. 6% were treated with aminosalicylate, glucocorticoids and immunosuppressant. After medicine treatment, 6.8% of the patients relieved, 62.7% turned better, 28.8% were ineffective, 1.7% died, 55.9% recurd. And 21.2% were treated by surgical operation. A total of 42.4% of patients with CD had histroy of surgical operation and 9.1% were carried on more than one surgical operation.ConclusionsActive state patients with CD had high rate of complications, low extraintestial manifestatons, rare rate of dysplasia and canceration, and high rates of surgical operation and recurring.
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Objective To investigate the role of cyclooxygenase-2 (COX-2) in sinusoidal capillarization in liver cirrhotic rats. Methods The SD rats were intraperitoneally injected with carbon tetrachloride (CCl4) twice a week for 8 weeks to induce liver cirrhosis. The rats were randomly divided into three groups: normal control group (n= 10), model control group (n= 15) and rofecoxib treated group (received 10 mg/kg of rofecoxib daily, n = 15). Liver histopathology was examined by light microscopy, and sinusoidal ultrastructure was observed by transmission electron microscopy. Furthermore, the level of basement membrane proteins (collagen type Ⅳ, laminin) and their localizations in liver were determined by Western blotting and immunohistochemistry, respectively, and the microvessel density was detected following vWF (yon Willebrand factor) immunolabeling on liver tissue sections. Results Fibrotic areas were reduced in rofecoxib treated group compared with that in model group (30.7±8.9 vs 23.5±6.5,P<0. 05). The light and electron microscopy showed that the pathologic changes including loss or reduction in number of sinusoidal endothelial fenestrate, the accumulation of extracellular matrix in the Disse's space and development of subendothelial basal lamina (basement membrane formation) were more severe in model group than those in rofecoxib treated group. Compared with model group, administration of rofecoxib resulted in significant decrease in microvessel density (11.3 ± 1.6 vs. 6.4 ±0. 7, P<0. 01). Rofecoxib could significantly decrease the expression of type Ⅳ collagen and laminin at protein levels (3.0±0.5 and 3.0±0.5, respectively) when compared with model group (3.8±0.4 and 3.7±0. 5, respectively). Conclusion The results indicate that early administration of rofecoxib may reduce sinusoidal capillarization.
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Objective To study the tumorigenetic characteristics of side population (SP) cells and their exsistence in gastric cancer cell lines and tissues.Methods Flow cytometry and the DNA-binding dye Hoechst 33342 staining were used to analyze sorted SP cells in 3 gastric cancer cell lines(SGC-7901,BCIC-823 and MKN28).SP cells and non-SP cells(main population cells)isolated from SGC-7901 cell line weresubcutaneously injected into 36 nude mice with 500,5000 or 50 000 cells per mouse,respectively.Thetumorigenity was observed 8 weeks after injection.The expression of ATP-binding cassette sub-family Gmember 2(ABcG2)mRNA in three gastric cell lines and its level in gastric cancer tissues were detected by real-time PCR and immunostaining.respectively.Results The proportion of SP cells was accounted for 1.0%in SGC-7901 cells and 1.3%in BGc-823 cells.No SP cell was found in MKN28 cells.As low as 500 SP cells or 50 000 non-SP cells could initiate tumors in mice.The expression of ABcG2 mRNA was higher in SGC-7901(O.162)and BGC-823(O.096)cell lines than that in MKN28 cell line(0.005).The value of AtCG2 mRNA/beta-aetin mRNA and protein in gastric cancer tissues was different from those in gastritis tissues.Condasimm SP cells have strong ability of tumorigenesis compared with non-SP cells.The expression of ABCG2 is found in gastric cancer tissues and part gastritis tissues.The more the SP cells in gastric cell lines,the higher the expression of ABCG2.
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Objective To investigate the relationship between HBV-DNA level during the course and progress tO cirrhosis in patients with chronic hepatitis B.Methods From 2001 to 2007,a total of 239 chronic hepatitis B patients confirmed by liver biopsy were followed up for a median time of 28 months.HBV-DNA level was measured at baseline and end point.Results Those who progressed to cirrhosis were older and with higher HBV-DNA levels at the end point.Kaplan-Meier analysis showed that the higher the HBV-DNA level at end point,the higher the risk to cirrhosis(X2=11.736,P=0.019).There was no difference between patients with and without cirrhosis at baseline of HBV-DNA level(P=0.531).The Cox regression indicated that the independent risk factors of cirrhosis were as followings:HBV-DNA level at the end point,stage of fibrosis,hepatitis B e antigen negative and γ-glutamyl transpeptidase at entry with relative risk ratio of 1.898,1.918,8.976 and 1.006,respectively.Conclusion HBV-DNA level is correlated with progress to cirrhosis in patients with chronic hepatitis B.
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Objective To investigate the expression of integrin avβ3 in both of hepatic stellate cell (HSC)and fibrotic liver tissue,and to demonstrate whether integrin avβ3 is a phenotypical receptor of activated HSC.Methods HSC were isolated from Sprague-Dawley rats and activated by prolonged cultural.The rats were injected with 175 mg of thioacetamide twice a week for 12 weeks to induce liver fibosis.The expressions of avβ3 and a-SMA were identified by immunocytochemical staining.The expression of avβ3 in HSC,normal and fibrotic tissues were examined by real-time PCR and Western blot.Results The expression of avβ3 in activated HSC were up regulated and levels of mRNA and protein were increased to 18 and 5.2 folds on day 14 compared with day 1 and were also higher than control(t=2.39,P＜0.05;t=2.74,P＜0.05).The immunochemistry staining showed that integrin avβ3 was expressed on membrane of activated HSCs.The avβ3 and a-SMA were expressed in portal vein in normal liver,but were also expressed in portal and fibrotic.Conclusions The integrin avβ3 is up-regulated in activated HSC both in vitro and in vivo.It is a phenotypical receptor of activated HSC which involved in liver fibrosis.
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<p><b>OBJECTIVE</b>To explore the relationship between plasma levels of carbon monoxide and blood-brain permeability in cirrhotic rats.</p><p><b>METHODS</b>We measured mean arterial pressure (MAP), heart rate, plasma levels of carbon monoxide and amount of Evans blue in the brain tissue taken as the index of blood-brain permeability in cirrhotic rats (n=10) and controls (n=10).</p><p><b>RESULTS</b>Cirrhotic rats showed significant increases in plasma carbon monoxide and amount of Evans blue in brain tissue compared with controls [(18.37 +/- 1.79) micromol/L,(18.52 +.- 1.39) ng/mg vs (10.27 +/- 1.21) micromol/L, (15.08 +/- 1.06) ng/mg; P< 0.01]. Carbon monoxide levels in cirrhotic rats correlated positively with blood-brain barrier permeability and inversely with MAP (r=0.72, P< 0.01; r= -0.67, P< 0.05).</p><p><b>CONCLUSIONS</b>Carbon monoxide, involving in the occurrence of hypotension and the increase of blood-brain barrier permeability, may play a role in the development of hepatic encephalopathy.</p>
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Animals , Male , Rats , Alanine Transaminase , Blood , Aspartate Aminotransferases , Blood , Blood Pressure , Physiology , Blood-Brain Barrier , Physiology , Capillary Permeability , Physiology , Carbon Monoxide , Blood , Carbon Tetrachloride , Heart Rate , Physiology , Liver Cirrhosis, Experimental , Blood , Rats, Sprague-Dawley , gamma-Glutamyltransferase , BloodABSTRACT
<p><b>OBJECTIVE</b>In order to improve the cognition and early diagnosis of primary biliary cirrhosis (PBC), we investigated clinical and pathological characteristics of PBC.</p><p><b>METHODS</b>Clinical data of 37 PBC patients together with pathological findings of 20 PBC patients were reviewed.</p><p><b>RESULTS</b>Among the 37 patients, 35 were women and the mean age at diagnosis was (53.4 8.9) years. The most frequent clinical presentations were jaundice (70.3%), fatigue (70.3%), and pruritus (56.8%). Serum glutamyl transpeptidase (GGT) and alkaline phosphatase (ALP) levels were markedly elevated in all patients (P50 was 467.50 U/L and 424.00 U/L, respectively). Among the 37 cases, 35 (94.6%) had total bile acids elevation, 32 (86.5%) had serum cholesterol elevation and 32 (86.5%) serum IgM elevation, 34 (91.9%) were positive for anti-mitochondrial antibody (AMA) and/or AMA-M2. Liver biopsy in 20 PBC patients mainly found: fibrosis in 17 cases (85%), interlobular bile duct lesions in 13 (65%), lobular mononuclear inflammation in 13 (65%), lymphocytic hepatocellular piecemeal necrosis in 10 (50%), and bile pigment accumulation in 9 (45%). The results of nonparametric test showed that GGT was related with pathological stage (P=0.002) and interlobular bile duct lesions (P=0.01).</p><p><b>CONCLUSIONS</b>PBC is mostly found in middle-aged women. Accurate and prompt diagnosis of PBC should be based on the clinical presentation, biochemical and immunological indexes, and hepatic pathological changes. The level of GGT may partly reflect the severity of the histological lesions</p>
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Female , Humans , Male , Middle Aged , Bile Acids and Salts , Biopsy , Fatigue , Liver Cirrhosis, Biliary , Pathology , Mitochondria , Portal System , Pruritus , gamma-GlutamyltransferaseABSTRACT
<p><b>OBJECTIVE</b>To assess the effectiveness of an educational intervention program in sexually transmitted disease (STD) and condom-related knowledge and promoting condom use among STD patients.</p><p><b>METHODS</b>STD clinic patients were randomly assigned to three groups (A, control group; B, video viewing group; group C, video viewing plus education with talk) after administered with baseline questionnaire. Each patient was then scheduled to complete the questionnaire regarding the change of STD knowledge and use of condom in two weeks and 3 months.</p><p><b>RESULTS</b>Six hundred and forty-eight patients entered the study. More than half of the patients were aware of STD-related knowledge. However the rate of condom use was very low. When having intercourse with the casual sexual partners the percentage of condom use was only 20%. Seventeen per cent of the patients knew nothing about how to use condom correctly. Follow up results showed that the education program increased the knowledge about STD to some degree. However the percentage of condom use in the latest sexual intercourse increased greatly both in intervention and control groups, but more in intervention groups (P < 0.05).</p><p><b>CONCLUSION</b>STD education should be focused on advocating and correcting condom use.</p>
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Adult , Female , Humans , Male , Condoms , Health Knowledge, Attitudes, Practice , Health Promotion , Methods , Patient Education as Topic , Methods , Sexually Transmitted Diseases , Videotape RecordingABSTRACT
Purpose To explore the clinical characteristics of small intestinal tumors and its diagnosis. MethodsRetrospective analysis of cases. ResultsLdomyoma was most common among the benign small intestinal tumors(12/17),adenocarcinomawas most common among the malignant small intestinal tumors(45/ 76). The feature of the small intestinal tumors could be hemorrhage, abdominal pain, abdominal mass, fever and jaundice. Upper gastrointestinal tract barium oontrast study was performed in 28 cases,and the results of 21 cases were positive. Mesenteric arteriography was performed in 9 cases. It was confirmed by surgery that the results matched with the lesions. ConclusionsMesenteric arteriography is of great value in diagnosis of the small intestinal tumors endoscope retrograde cholangiopancreatography(ERCP), CT are also helpful.
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Objective To establish a new Helicobacter pylori(H.pylori) infection model which is more stable and to find the relationship between H. ~pylori infection and gastric carcinogenesis induced by nitrosourea. Methods A total of 94 Balb/c mice were divided into four groups. Two groups of mice were inoculated with H.pylori, among which one group of mice received continuous N-methyl-N-nitrosourea(MNU) administration via the drinking water. The third group were only given MNU, and the forth as control. After 36 weeks, all the mice were ~sacrificed . The infection of H.pylori in gastric mucosa of the mice was analyzed by rapid urease test, ~Giemsa staining and culture. The histopathological changes in the gastric mucosa of mice were assessed in ~H-E stained sections. Results The total infection rate of H.~pylori in Balb/c mice was 93.9%. All the mice in group with H.pylori infection alone had chronic gastritis, in which 20.0% was atrophic gastritis. In the group of mice with H.pylori infection and MNU intake, all had chronic gastritis, and among them 23.1% had atrophic gastritis, 42.3% corpus dysplasia and 57.7% antrum dysplasia. In this group, two mice (7.1%) with low differentiated adenocarcinoma were found. Compared with control group, two groups with H. ~pylori infection only and H. ~pylori plus MNU intake reached statistical difference in the view of inflammatory latitude (P
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PurposeTo establish sensitive and reliable serum marker on liver fibrosis for early clinical diagnosis. MethodsSerum hyaluronate acid,type Ⅲ procollagen and type Ⅳ procogen of 48 patients with viral hepatitis were analyzed by radioimmunoassay and ELISA respectively. ResultsThe serum HA, PCⅢ and PCⅣ level increased gradually with the progress of the disease. Their levels were correlated distinctly with the index of hepatic activity of inflammation and that of fibrosis.ConclusionsThe value of serum HA,PCⅢ and PCⅣ in predicting liver fibrosis will be enhanced ff the effect of inflammation could be excluded.
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Objective To investigate the role of cyclooxygenase 2 (COX 2) in the forming of portal hypertension and whether a selective COX 2 inhibitor can reduce the portal hypertension or not. Methods Cirrhotic Sprague Dawley rat was induced by carbon tetrachloride (CCl 4) intraperitoneally for 8 weeks. The animals were divided into three groups: 10 normal rats served as control group; the other 15 rats, which received CCl 4 intraperitoneally twice a week and rofecoxib (10 mg/kg) by gavages daily, served as treatment group; another 15 rats, which were induced cirrhosis by CCl 4 but given placebo (saline solution ) instead of rofecoxib, served as placebo group. After 8 weeks of CCl 4 induction, portal pressure was measured, and the levels of thromboxane B 2 (TXB 2), and prostaglandin (PG)E 2 in the liver tissues were determined by enzyme immunoassay. Furthermore, liver histopathological analysis was performed in H E and Masson's trichrome staining sections. Results Portal pressure in the rats of rofecoxib group was significantly decreased compared to that in the placebo group [(11.95?1.05) mm Hg vs. (13.45?1.15) mm Hg; P
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Objective To investigate the effects of glycyrrhizin on gene expression of transforming growth factor(TGF)-?signaling pathway of hepatic stellate cell(HSC)in mice by gene microarray tech- nique.Methods The HSCs were isolated from mice and cultured in vitro.Then the mice were divided into control group,TGF-?_1 group(5 ng/ml) or TGF-?_1(5 ng/ml)combined with glycyrrhizin(100?mol/L) group.The cells were collected after 10 hours to extract RNA.A cDNA microarray(GEArray~(TM) Q) targeting TGF-?/BMP signal transduction was used to screen the genes which showed significant changes in expression of TGF-?pathway of HSC by glycyrrhizin.Results The microarray analysis showed that 16 genes(16.7%),such as Smad2,Smad3,Smad7,CoL3A1,CoL1A2,and PAI-1,were upregulated by TGF-?_1 and then down-regulated by glycyrrhizin.Five genes(5.2%)(including BMP7,IGFbp3 and etc.)downregulated by TGF-?_1,were then up-regulated by glycyrrhizin.Finally,2 genes upregulated by TGF-?_1 were then up-regulated predominantly by glycyrrhizin in HSC(T?R2,betaglycan).Changes in some genes,such as Smad2 Smad3,Smad7,were further confirmed to be coincided with cDNA microarray by semi-quantitative RT-PCR.Conclusions The anti-fibrosis mechanisms of glycyrrhizin may be through to interference of TGF-?signaling pathway,decrease the synthesis and increase the degradation of collagen.
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Objective To study the expressing status of antisense tissue inhibitor of metalloproteina se-1(TIMP-1) in hepatic stellate cells (HSC) constructed in vitro, and to eval u ate the effects on the production of type Ⅰ and Ⅲ collagens secreted by activated rat HSC. Methods HSC were extracted from normal rat liver by pronase and co llagenase digestion and purified by centrifugal elutriation, and were cultured pla stic until they were activated to a myofibroblastic phenotype after 7-10 days. RT-nest-PCR and gene recombinant techniques were used to construct the rat ant isense TIMP-1 expression plasmid which can express in eukaryotic cells, and seg uenced after being counstructed. The expressing plasmid and the pcDNA3 empty pla smid were transfected into HSC by Effectene reagent separately. The cells were sel ected after growing in DMEM containing 400 ?g/ml G418 for 3 to 4 weeks. Exp ression of TIMP-1 in HSC was d etermined by Northern blot and Western blot. We tested the interstitial collagen ase activity in culture media with FITC-labled type Ⅰ collagen as substrate. U ltimately, we quantified the typeⅠ and type Ⅲ collagen in HSC by Wester n blot. Results The exogenous antisense TIMP-1 recombinant plasmid could block the expression of TIMP-1 greatly, while there were not the same outcome i n pcDNA3 empty plasmid g roup and non-transfecting control group. The ratio of TIMP-1/GAPDH was 0.67, 2 .41 and 2.97 respectively at mRNA level( P