BACKGROUND:A large number of experiments have confirmed that bone marrow mesenchymal stem cells can differentiate into hepatocytes under the induction of cytokines and specific micro-environment, and have been widely used in clinical alternative treatment for terminal liver disease, but the optimal inducing conditions are unclear.
OBJECTIVE:To explore the possibility and validity of differentiation of rat bone marrow mesenchymal stem cells into hepatocytes with a culture system containing salidroside and cholestatic rat serum in vitro.
METHODS:Bone marrow mesenchymal stem cells were isolated by plastic adherence from the whole bone marrow of health rats, and cellphenotypes were identified using the flow assay;cholestatic serum was prepared by common bile duct ligation. Passage 3 bone marrow mesenchymal stem cells were randomly divided into three groups for in vitro induction by the different culture systems:blank control group:basic medium plus 5%cholestatic serum;salidroside group:basic medium plus 5%cholestatic serum plus 30 μmol/L salidroside;positive control group:basic medium plus 5%cholestatic serum plus 20 μg/L hepatocyte growth factor. Changes of cellmorphology during culture time were observed in each group, reverse transcription-PCR assay and western blot assay were used to expression of hepatocyte-specific proteins.
RESULTS AND CONCLUSION:The bone marrow mesenchymal stem cells highly expressed CD90, CD105, but did not express CD45, CD14, CD34, and CD79a. Polygonal and binucleate cells appeared in the three groups during the procedure of induction. The mRNA and protein expression of alpha-fetoprotein and albumin emerged in the three groups on the 7th day;in the same period, the lowest expression ratio was in the blank control group (P<0.05), while there was no significant difference between the salidroside and positive control groups (P>0.05). Combination of salidroside and cholestatic serum can effectively induce bone marrow mesenchymal stem cells differentiating into hepatocytes.

Hepatic Veins ; physiopathology ; Humans ; Hypertension, Portal ; physiopathology ; Portal Pressure

Arteriovenous Fistula ; Humans ; Hypertension, Portal

Objective To investigate the effect of Salidroside in inducing apoptosis of rat hepatic stellate cells (HSC) stimulated by acetaldehyde and to observe the changes of c- Jnk N- terminal kinase (JNK) activity.Methods HSC stimulated by acetaldehyde were cultured in vitro and were treated with different concentrations of Salidroside.Apoptotic rate was analyzed by flow cytometry and the activity of phosphorylating JNK was measured by Western blot method.Results Salidroside in different concentrations (1.0,1.5,2.0 mg/mL) suppressed the activity of JNK in a dose- effect manner.Average light density was 35.8? 3.4,24.9? 2.7 and 3.4? 0.9 in Salidroside groups, which differed from that in acetaldehyde group( 48.6? 4.8; P

BACKGROUND:Matrix metaloproteinase-1 can degrade extracelular matrix, which is mainly colagen type I, and has the potential to reverse fibrosis tissue. OBJECTIVE:To construct the recombinant adenovirus vector containing human matrix metaloproteinase-1 (hMMP-1) gene with GatewayTM Clone Technology, and observe the capacity of degrading colagen type IIIin vitro. METHODS: The gene hMMP-1 was amplified by using PCR from the pcDNA3.1 plasmid and was cut down by the double endonuclease. The linear gene fragment was connected to the entry vector pENTERTM 1A. Then the entry clone and the destination vectors pJTI? R4 Dest CMV-N-EmGFP pA Vector recombined using the LR reaction to form the expression clone pAd-hMMP-1-eGFP. The linear pAd-hMMP-1-eGFP cut down by endonucleasePac I was transfected into HEK293A cels to packaging the Ad-hMMP-1-eGFP. The transfected situation was observed under a fluorescence microscope, the target protein expression was detected by western-blot assay and RT-PCR. Cels can be divided into three groups: blank control group: HEK293A cels, AD-EGFP group: HEK293A cels were infected by Ad-eGFP, AD-HMMP1-EGF group: HEK293A cels were infected by Ad-hMMP1-eGFP and colagen type III. The content of colagen type III was detected by ELISA kits after 24, 48 and 72 hours. RESULTS AND CONCLUSION: It was confirmed that the entry vector and the destination vector both contained hMMP-1 target gene by restriction analysis and sequencing. The green fluorescent protein was observed in the 293A cels transfected by the Ad-hMMP-1-eGFP at 4 days. The fluorescence intensity was the highest at 10 days. The virus was colected at 12 days, the viral titer was determined as 4.84 × 1010 PFU/mL, the target protein was efficient expressionvia western-blot assay. Blank control group and AD-EGFP group had no obvious change of colagen content with the extension of time. The rate of colagen degradation in AD-HMMP1-EGFP group was 24%, 56% and 81% respectively at 24, 48, 72 hours. AD-HMMP1-EGFP group degraded colagen significantly compared with the other two groups (P < 0.01). The recombinant adenovirus vector containing hMMP-1 was successfuly constructed by using the Gateway technology, this method was more efficient and specific than with the traditional methods. The hMMP1 degraded colagen type III significantlyin vitro.

Liver failure is a clinical syndrome with severe disorders of liver cells for biosynthesis, detoxication, excretion, and biological transformation, which presents with coagulation disorders, jaundice, hepatic encephalopathy, and ascites. Liver failure progresses rapidly, so the prediction of clinical outcome is significant for the diagnosis and treatment. In recent years, there have been numerous reports on the prediction of clinical outcome in patients with liver failure. The study and application of serological and comprehensive models are reviewed, which provides a reference for the rational therapy for liver failure patients.

ObjectiveTo analyze the efficacy of endoscopic histoacryl injection in the treatment of gastric variceal bleeding caused by regional portal hypertension. MethodsThe endoscopic features and efficacy of endoscopic histoacryl injection were examined and compared in two groups of patients admitted to our hospital from June 2012 to December 2012. One of the groups included 6 patients with gastric variceal bleeding caused by regional portal hypertension and the other group included 6 patients with gastric variceal bleeding caused by hepatitis B cirrhosis-related portal hypertension. Between-group comparison of categorical data was made by Fisher′s test. ResultsIn patients with regional portal hypertension, five of them had severe isolated gastric varices (IGV) and one had severe IGV with mild esophageal varices. All six patients with hepatitis B cirrhosis-related portal hypertension had severe IGV and the endoscopic features were similar to those of patients with regional portal hypertension. Significant differences were observed between the group with regional portal hypertension and the group with hepatitis B cirrhosis related portal hypertension in short-term response rate (1/6 vs 6/6, P=0.015) and long-term response rate (0/6 vs 5/6, P=0.015). ConclusionThe gastric varices caused by regional portal hypertension has a fast progression rate and a high bleeding risk. The efficacy of endoscopic histoacryl injection in patients with this type of gastric varices is poor.

Portal hypertension is a common clinical disease and brings a series of complications including the formation of gastrointestinal varicose veins, ascites, hepatic encephalopathy, and abdominal varicose veins. Most of these complications are related to the opening of collateral circulation after the increase in portal venous pressure. On one hand, collateral circulation helps to alleviate the high portal venous pressure, and on the other hand, it brings related complications to patients. This article reviews recent reports and studies on collateral circulation related to portal hypertension, in order to increase our knowledge of collateral circulation in portal hypertension and improve clinical diagnosis and treatment of such disease.

There are not many studies on chronic hepatitis C complicated by autoimmune hepatitis, and up to now, the clinical diagnosis and treatment of such diseases still face many difficulties. Although related articles put forward some recommendations, there are no standard guidelines for diagnosis and treatment, and clinical physicians need to provide treatment for these patients based on their personal experience. This article summarizes related articles on chronic hepatitis C complicated by autoimmune hepatitis in order to provide help to clinical physicians when they face similar clinical problems in the future.

ObjectiveTo observe the effect of continuous blood purification (CBP) on serum inflammatory mediators in patients with severe acute pancreatitis (SAP) and multiple organ dysfunction syndrome (MODS). MethodsSixty-five SAP patients with MODS who were treated in General Hospital of Chengdu Command Area of Chinese PLA from April 2008 to December 2013 were enrolled and divided into two groups. The 33 patients in the control group received comprehensive internal medicine treatment, and the 32 patients in the treatment group received comprehensive internal medicine treatment and CBP. Changes in APACHE II score, MODS score, and the serum levels of tumor necrosis factor α (TNFα), C-reactive protein (CRP), interleukin 6 (IL-6), IL-18, platelet-activating factor (PAF), and nitric oxide (NO) after treatment were observed. Independent-samples t test was applied for comparison of continuous data between the two groups, and paired t test was applied for before-after comparison within the same group; chi-squared test was applied for comparison of categorical data between the two groups. ResultsIn both groups, APACHE II score, MODS score, and the serum levels of TNFα, CRP, IL-6, IL-18, PAF, and NO decreased significantly after treatment (all P＜0.05), and the treatment group had significantly greater decreases in these values than the control group (all P＜0.001); the survival rates in the treatment group and the control group were 90.6% (29/32) and 78.8% (26/33), respectively, with no significant difference between the two groups (χ2=1.749, P=0.186). ConclusionIn SAP patients with MODS, CBP can effectively clear the serum inflammatory mediators to block systemic inflammatory response and improve organ function, and, therefore, it is an effective method to treat SAP.