The establishment and perfection of the assessment system for chronic liver diseases have a guiding significance for clinical diag-nosis and treatment.Hepatic venous pressure gradient (HVPG)is of great significance in the progression of chronic liver diseases,and it is the only pathophysiological index for treatment independent of etiology.The current methods for evaluation of chronic liver diseases are sim-ply described,and the application of HVPG in evaluation of chronic liver diseases is systematically summarized,including predicting the se-verity of liver cirrhosis,variceal bleeding risk and outcome of portal hypertension,development of cirrhotic ascites,efficacy of drugs for re-ducing portal hypertension and antiviral drugs,and postoperative outcome of liver cancer.It is thought that HVPG measurement plays an im-portant role in the assessment of the progression and prognosis of chronic liver diseases and is an effective predictor of outcome.

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.

Arteriovenous Fistula ; Humans ; Hypertension, Portal

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.

Objective To study the effect of p38MAPK on the activity and c-myc protein expression in rat acetaldehyde-induced hepatic stellate cell(HSC),and to investigate the alcoholic liver fibrosis related mechanism.Methods The different concentrations of SB203580 as the p38 specific blocker was adopted to conduct the intervention on rat acetaldehyde-induced HSC.The cellular mor-phological change was observed by the microscope.The cell proliferation was detected by MTT,the cell cycle was analyzed by flow cytometry(FCM),and the expression of c-myc protein was examined by the SABC method.Results (1)after acetaldehyde stimula-tion,HSC was increased in size and proliferated rapidly,but with the added SB203580 concentration increase,the cellular prolifera-tion was slowed down,the cells size was diminished and the deformed cells were increased.(2)The proliferation of acetaldehyde-in-duced HSC was inhibited by different doses of SB203580,and the higher concentration has the more significant inhibiting effect.(3) With the SB203580 concentration increase,the cells at the phase G0 and G1 were increased,while the cells at the phase S were de-creased,at the same time the expression positive rate of c-myc protein was decreased.Conclusion Blocking p38MAPK pathway ac-tivity could inhibit the proliferation of acetaldehyde-induced HSC,which may be related to the down-regulation of C-myc protein ex-pression and blocking the DNA synthesis in cells entering from G0/G1 phase to S phase.

The performance of transjugular intrahepatic portosystemic shunt (TIPS) has two key procedures: (1) portal vein branch puncturing, and (2) the correct judgment of the safety of the puncture site. The portal vein branch puncturing is the most important and difficult step for a successful TIPS procedure. Therefore, to find and to establish an proper access to the portal vein is critical. Nowadays, in clinical practice several imaging techniques have been used to localize the portal vein, such as magnetic resonance imaging, sonography, fluoroscopy, arteriography and computed tomography. This article aims to make a general review on these invasive and non - invasive localization techniques when a successful performance of TIPS is expected.

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

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

Objective To investigate the effects of PD98059, the specific blocking agent of mitogen extracellular signal responsive kinase, on the cell cycle, cell proliferation, type Ⅰ collagen's secretion and transforming growth factor(TGF)? 1mRNA expression of rat hepatic stellate cells (HSC) stimulated by acetaldehyde. Methods Rat HSC stimulated by acetaldehyde were incubated with different concentration of PD98059. Cell cycle was analysed by flow cytometry. Cell proliferation was assessed by MTT colorimetric assay. Type Ⅰcollagen of culture medium was detected by enzyme linked immunoadsordent assay (ELISA). TGF? 1mRNA expression were examined by RT PCR. Results 20,50,100 ?mol/L PD98059 could significantly inhibit proliferation and induced G0/G1 phase arrest of HSC stimulated by acetaldehyde in a does dependent manner. 50,100 ?mol/L PD98059 could markedly inhibit type Ⅰcollagen's secretion and TGF? 1mRNA expression of HSC stimulated by acetaldehyde. Conclusion The extracellular signal regulated kinase signal transduction passway regulates the cell proliferation, type Ⅰcollagen's secretion and TGF? 1mRNA expression of rat HSC stimulated by acetaldehyde,which might be partly related to its regulative effect on the cell cycle.

OBJECTIVE: To evaluate the efficacy and safety of Dinggui Oil Capsule in treating irritable bowel syndrome (IBS) with stagnation of qi and cold. METHODS: A prospective, randomized, placebo-controlled, double-blind clinical study was undertaken. One hundred and ninety-eight patients with IBS and syndrome of stagnation of qi and cold were randomly divided into high-dose Dinggui Oil group (DGO-H, 1.2 g, 3 times daily; n=66), low-dose Dinggui Oil group (DGO-L, 0.8 g, 3 times daily, n=66), and placebo group (placebo, 5.0 g, 3 times daily, n=66). Patients in the three groups were all treated for 2 weeks. RESULTS: The total significant effective rates for IBS were 54.1%, 28.8% and 21.9% in the DGO-H, DGO-L, and placebo groups, and the total effective rates for the syndrome of stagnation of qi and cold were 54.1%, 25.8% and 23.4% in the three groups, respectively. Dinggui Oil Capsule showed a higher efficacy than the placebo in relieving the abdominal pain (P<0.01). No adverse effects were found in this trial. CONCLUSION: Dinggui Oil Capsule is effective and safe in relieving abdominal pain due to IBS with stagnation of qi and cold.