EGF Suppresses the Initiation and Drives the Reversion of TGF-ß1-induced Transition in Hepatic Oval Cells Showing the Plasticity of Progenitor Cells.

Transforming growth factor-ß1 (TGF-ß1) induces hepatic progenitors to tumor initiating cells through epithelial-mesenchymal transition (EMT), thus raising an important drawback for stem cell-based therapy. How to block and reverse TGF-ß1-induced transition is crucial for progenitors' clinical application and carcinogenic prevention. Rat adult hepatic progenitors, hepatic oval cells, experienced E-cadherin to N-cadherin switch and changed to α-smooth muscle actin (α-SMA) positive cells after TGF-ß1 incubation, indicating EMT. When TGF-ß1 plus EGF were co-administrated to these cells, EGF dose-dependently suppressed the cadherin switch and α-SMA expression. Interestingly, if EGF was applied to TGF-ß1-pretreated cells, the cells that have experienced EMT could return to their epithelial phenotype. Abruption of EGF receptor revealed that EGF exerted its blockage and reversal effects through phosphorylation of ERK1/2 and Akt. These findings suggest an important attribute of EGF on opposing and reversing TGF-ß1 effects, indicating the plasticity of hepatic progenitors.

Emodin improves lipopolysaccharide-induced microcirculatory disturbance in rat mesentery.

OBJECTIVE: Sepsis is a systemic inflammatory response syndrome. Emodin is a major ingredient of Rheum Palmatum, a Chinese herb that is widely used in China for treatment of endotoxemia-related diseases. This study intended to examine the effect of Emodin on LPS-induced rat mesenteric microcirculatory disturbance and the underlying mechanisms. METHODS: The male Wistar rats received LPS (5 mg/kg/hr) for 90 min, with or without administration of Emodin (10 mg/kg/hr) by enema 30 min before (pre-treatment) or after (post-treatment) LPS infusion, and the dynamics of mesenteric microcirculation were determined by inverted intravital microscopy. Expression of adhesion molecules and TLR4, NF-κB p65, ICAM-1, MPO, and AP-1 in mesentery tissue was evaluated by flow cytometry and Western-blot, respectively. RESULTS: Pre or post-treatment with Emodin significantly ameliorated LPS-induced leukocyte emigration, reactive oxygen species production and albumin leakage, and the expression of TLR4, NF-κB p65, ICAM-1, MPO and AP-1 in mesentery. CONCLUSIONS: These results demonstrate the beneficial role of Emodin in attenuating the LPS-induced microcirculatory disturbance, and support the use of Emodin for patients with endotoxemia.

[Serum sodium concentration profile for cirrhotic patients and its effect on the prognostic value of the MELD score].

To analyze the characteristics of serum sodium in decompensated cirrhosis and evaluate the prognostic ability of the model for end-stage liver disease (MELD) in Na-containing models. Patients diagnosed with decompensated cirrhosis at our hospital were enrolled for study between June 2005 and October 2010. Patients were classified among three groups, according to serum sodium concentration: less than 125 mmol/L, 125 to 135 mmol/L, and more than 135 mmol/L. Mortality rates among the three groups were compared by Kaplan-Meier survival analysis. In addition, the different serum sodium concentrations were analyzed for correlations between Child-Pugh score and complication incidence rates of portal hypertension. The area under the receiver operating characteristic (ROC) curve (AUC) was used to compare the predictive abilities of MELD, MELD-Na, and the integrated (i) MELD scores for 3-month, 6-month and 1-year mortalities. A total of 467 patients were analyzed, and 50.54% had hyponatremia ( less than 135 mmol/L). Sodium concentration was correlated with mortality, and Kaplan-Meier survival analysis indicated that mortality was significantly higher in each subgroup with lower sodium concentration (all, P = 0.000). Likewise, sodium concentration decreased in conjunction with increased severity of decompensation, as classified by Child-Pugh scoring (sodium: A more than B more than C; mortality: A less than B less than C). With the exception of digestive tract bleeding, complication incidence rates of hepatic encephalopathy, ascites, spontaneous bacterial peritonitis, and hepatorenal syndrome increased when sodium concentration decreased. For predicting 3-month mortality, the AUC scores of MELD were not significantly different from the MELD-Na and iMELD scores (P more than 0.05). For predicting 6-month and 1-year mortality, the AUC scores of MELD-Na and iMELD were significantly higher than those of MELD (P less than 0.05). Hyponatremia is correlated with mortality and complications in decompensated cirrhosis patients. Incorporation of Na into the MELD may enhance it's prognostic ability.

Primary isolated hepatic oval cells maintain progenitor cell phenotypes after two-year prolonged cultivation.

BACKGROUND & AIMS: Although expandable hepatic progenitors provide renewable cell sources for treatment of hepatic disorders, long-term cultivation of hepatic progenitors may affect proliferation and differentiation abilities, and even initiate the formation of malignant cancer stem cells. This study aims to determine characteristics of primary cultured hepatic oval cells after prolonged cultivation in vitro. METHODS: Hepatic oval cells isolated from rats fed with a choline-deficient, ethionine-supplemented diet were continuously propagated every 5-7 days, to 100 passages over two years. Hepatocytic differentiation was induced by sodium butyrate and characterized using western blot, periodic acid Schiff assays, albumin secretion and urea production. Proliferation capacity was evaluated using growth-curve and cell-cycle analysis; anchorage-independent growth and tumorigenicity were determined using soft agar and xenograft assay. RESULTS: After 2 years of serial passages, hepatic oval cells with typical epithelial morphology continuously expressed OV-6, BD-1, BD-2, and Dlk as markers for hepatic progenitors, cytokeratin 19 as a cholangiocyte marker, and alpha-fetoprotein and albumin as hepatocyte markers. Furthermore, sodium butyrate could induce these cells to become glycogen-storage cells with the functions of albumin secretion and ureagenesis from ammonia clearance, indicating hepatocytic differentiation. Although proliferation slightly accelerated after the 50th passage, hepatic oval cells stayed diploid cells with features of chromosomal stability, which did not acquire anchorage-independent growth capacity and caused no tumor in immunodeficient mice, suggesting no spontaneous malignant transformation. CONCLUSIONS: Hepatic oval cells retain the progenitor cell features without spontaneous malignant transformation after prolonged cultivation, and thus may serve as an expandable cell source for future exploitation of stem cell technology.

Assessment of hepatic fibrosis by transient elastography in patients with chronic hepatitis B.

Hepatic ultrasonic transient elastography (FibroScan) is a new diagnostic method for the assessment of hepatic fibrosis. There are limited data available on its use as a follow-up tool for patients with chronic hepatitis B. In this study, 134 patients were enrolled. Hepatic fibrosis was evaluated by liver stiffness measurement and biopsy. The biopsy criteria of the Chinese Program of Prevention and Cure for Viral Hepatitis, Metavir classification, and the modified Chevalier's semiquantitative system were used for histological assessment. The liver stiffness value was correlated with fibrosis stage (r = 0.565, P < 0.001) and fibrosis semiquantitative score (r = 0.727, P < 0.001). The liver stiffness value of G2 was significantly higher than that of G1 within the same fibrosis stage for S1, S3, and S4, respectively. Three patients were graded as G1S1, and had moderate steatosis without distinct fibrosis in the portal area and lobule, while their liver stiffness values were higher than 6.2 kPa. Although belonging to the same fibrosis stage, for thinner thicknesses of the fibrous septa, the liver stiffness value and semiquantitative score were correspondingly lower. Liver stiffness values had a good correlation with hepatic collagen content. However, inflammatory activity and steatosis can influence liver stiffness values to some extent. Transient elastography may be useful as an ideal non-invasive post-treatment follow-up tool.

Glutamine synthetase as an early marker for hepatocellular carcinoma based on proteomic analysis of resected small hepatocellular carcinomas.

BACKGROUND: Hepatocellular carcinoma (HCC) is a highly malignant tumor with a poor prognosis. Because small HCCs possess most of the characteristics of early HCC, we investigated small HCCs to screen potential biomarkers for early diagnosis. METHODS: Proteins were extracted from 10 sets of paired tissue samples from HBV-infected small-HCC patients. The extracted proteins were well resolved by two-dimensional electrophoresis. These HCC-associated proteins were then identified by MALDI-TOF/TOF MS following image analysis. Western blotting and immunohistochemistry were used to assess glutamine synthetase (GS) and phenazine biosynthesis-like domain-containing protein (PBLD) expression in liver tissue. Enzyme-linked immunosorbent assays in 152 serum samples (from 49 healthy donors, 24 patients with liver cirrhosis, and 79 with HCC) were used to further assess the significance of GS clinically. RESULTS: Fifteen up-regulated and three down-regulated proteins were identified. Western blotting confirmed GS overexpression and decreased PBLD expression in liver tissue. Immunohistochemistry showed that GS was expressed in 70.0% (84/120) of HCCs and 35.8% (43/120) of nontumor tissues; PBLD was expressed in 74.2% (89/120) of nontumor tissues and 40.8% (49/120) of HCCs. The Chi-square test showed significant expression differences between HCCs and adjacent tissues. Consistent with this, serum GS levels in HCC patients were significantly higher than those in liver cirrhosis patients and healthy donors, while the latter two groups were also significantly different. In addition, a diagnostic cutoff value of 2.6 mg/ml was used for GS; it was elevated in 19 (76.0%) of 25 HCC patients with AFP

Pirfenidone inhibits carbon tetrachloride- and albumin complex-induced liver fibrosis in rodents by preventing activation of hepatic stellate cells.

1. Pirfenidone (PFD; 5-methyl-1-phenyl-2(1H)-pyridone) is an effective and novel agent with antifibrotic and anti-inflammatory properties. In the present study, we investigated the antifibrotic effects of PFD on experimental liver fibrosis models in rodents and the possible underlying molecular mechanisms. 2. Liver fibrosis was induced by carbon tetrachloride (CCl(4)) in BALB/c mice. Pirfenidone (250 mg/kg) and silymarin (50 mg/kg) were given to different groups of rats by gastric gavage for 4 weeks. Pirfenidone significantly attenuated fibrosis severity, as determined by histopathological scores and hydroxyproline levels in liver tissue, by 49.8 and 44.9%, respectively, compared with the CCl(4)-treated group. The antifibrotic effects of PFD were significantly greater than those of silymarin, as indicated by a decrease of 23.5 and 24.8% in histopathological scores and hydroxyproline levels, respectively. 3. Liver fibrosis was also induced by albumin antigen-antibody complex in Wistar rats, which were then treated with the same doses of PFD and silymarin for 8 weeks. Pirfenidone significantly reduced the degree of fibrosis compared with CCl(4)-treated rats (by 45.0 and 51.0% as determined by histopathological scores and hydroxyproline levels in liver tissue, respectively). The antifibrotic effects of PFD were comparable to those of silymarin. 4. The effects of PFD on the expression of extracellular matrix-associated genes in human hepatic stellate cells (the LX-2 cell line) were measured by real-time quantitative polymerase chain reaction. LX-2 cells were treated with or without 100 micromol/L or 1 mmol/L PFD for 24 h. Pirfenidone significantly inhibited the expression of a-smooth muscle actin and Type I collagen in 8 ng/mL transforming growth factor-beta1- or 5% fetal bovine serum-activated LX-2 cells in a dose-dependent manner. 5. In conclusion, the results of the present study demonstrate that PFD is effective in ameliorating fibrogenesis induced by CCl(4) in mice and by the albumin complex in rats. These effects were mediated mainly via inhibition of the activation of hepatic stellate cells, as well as antifibrotic actions (i.e. inhibition of collagen synthesis) of PFD.

[The effect of portal hypertension on prognosis in patients with decompensated liver cirrhosis].

OBJECTIVE: To evaluate the effect of portal hypertension on prognosis in patients with decompensated liver cirrhosis. METHODS: The clinical data of decompensated cirrhosis patients in our hospital, between 2003 and 2006, were retrospected and followed up. Model for end-stage liver disease (MELD) score and Child-Turcotte-Pugh (CTP) classification was calculated using the standard formula. Kaplan-Meier survival analysis was used to compare the mortality in subgroups ranked by the syndromes. Cox proportional hazards regression was used to evaluate the effect of the syndromes on prognosis. RESULTS: A cohort of 322 patients was admitted in this study at the end of the follow-up. The mortality of variceal bleeding, hepatic encephalopathy, a large volume ascites, spontaneous bacterial peritonitis, the type I and type II hepatorenal syndrome was 45.9%, 79.4%, 66.7%, 100%, 100% and 84.6% respectively. On the whole, the occurrence of all the syndromes was correlated with CTP classification and MELD score. Kaplan-Meier survival analysis showed that all of these syndromes, except for low to medium volume of ascites, significantly affected the survival rate (P<0.01). In Cox regression analysis, all the syndromes were the independent predictors of prognosis, the regression coefficient values of hepatic encephalopathy, spontaneous bacterial peritonitis, type I and type II hepatorenal syndrome, variceal bleeding and ascites were 0.973, 0.928, 0.935, 0.866, 0.464 and 0.369 respectively. CONCLUSIONS: The portal hypertensive syndromes have significant effect on the prognosis of the patients with decompensated cirrhosis, hepatic encephalopathy is the worst one.

[Analysis of primary diseases of 452 patients complicated by multiple organs dysfunction syndrome].

OBJECTIVE: To investigate the influence of the primary diseases on the outlook of multiple organs dysfunction syndrome (MODS). METHODS: The investigation was a multicentre study. The clinical data of 452 MODS cases from 20 III-level general hospitals in Northern China were analyzed with chi-square test and logistic regression analysis. The relationship of primary diseases, their distribution and prognosis of MODS was investigated. RESULTS: Mean age of patients (320 cases) who were suffering from a certain disease was (67.9+/-0.8) years old, which was higher than that of patients [132 cases, (46.9+/-1.7) years old] who did not have a primary disease (P=0.000). The mortality of patients with existing primary diseases was 56.2% (180/320), which was higher than that of the patients without a primary disease (31.1%, 41/132, P=0.000). The percent of occurrence of dysfunction of three organs was higher in patients who were suffering from some primary diseases was 74.38% (238 cases) than that of those without a primary disease (63.64%, 84 cases, P=0.022). In the patients who were suffering from cerebrovascular disease, cardiac insufficiency, chronic obstructive pulmonary disease (COPD), the incidence of complicating with dysfunction of three to four organs was respectively 63.4% (52/82), 67.1% (55/82), 69.1% (38/55), and were higher than those patients who did not have above-mentioned diseases [50.5% (187/370), 49.7% (184/370), 54.3% (201/370), all P<0.05]. Logistic regression analysis showed that cerebrovascular disease was the major risk factor. CONCLUSION: In patients whose age is over 67 years, and at the same time they are having a primary disease, the prognosis is poor. The mortality of MODS patients with hypertension, cerebrovascular disease, cardiac insufficiency, COPD, chronic renal insufficiency is higher, and cerebrovascular disease is the major risk factor in the prognosis of MODS patients.

[A clinical study on the diagnostic criteria of multiple organ dysfunction syndrome].

OBJECTIVE: To develop diagnostic criteria of multiple organ dysfunction syndrome (MODS) by a prospective and multi-center clinical investigation. METHODS: The data of 1087 MODS cases obtained from ICU of 37 hospitals from March 2002 to January 2005 in 11 provinces in China were analyzed in order to derive the diagnostic criteria of MODS. RESULTS: This MODS diagnostic criteria involved 7 organs. To diagnose MODS, the original cause of MODS should be identified, then there should be two or more organs showing signs of dysfunction. The criteria for organ dysfunction were as follows. (1) Cardiovascular system: SBP < 90 mm Hg (1 mm Hg = 0.133 kPa), MAP < 70 mm Hg, signs of shock, ventricular tachycardia, ventricular fibrillation, or myocardial infarction; (2) Respiratory system: oxygenation index < 300 mm Hg; (3) Nervous system: indifference, restlessness, lethargy, light coma, or deep coma, Glasgow score < or = 14; (4) Blood system: PLT < 100 x 10(9)/L; CT, APTT, and PT prolonged or shortened; positive plasma protamine paracoagulation; (5) Liver: TBIL > 20. 5 micromol/L, ALB < 28 g/L; (6) Kidney: Cr > 123.8 micromol/L, urinary volume < 500 ml/24 h; (7) Gastro-intestine: bowel sounds decreased or disappeared; retention in the stomach, or positive occult blood feces with dark stools or haematemesis; intraabdominal pressure (intravesical pressure) > or = 11 cm H2O (1 cm H2O = 0.098 kPa). Any organ function met with one of the above conditions was considered to have dysfunction. CONCLUSIONS: This diagnostic criterion of multiple organ dysfunction syndrome has been developed by this research, but it needs to accumulate experience by clinical practice and to revise the diagnosis criteria.

Down-regulation of transforming growth factor beta 1/activin receptor-like kinase 1 pathway gene expression by herbal compound 861 is related to deactivation of LX-2 cells.

AIM: To investigate the effect of herbal compound 861 (Cpd861) on the transforming growth factor-beta1 (TGF beta 1)/activin receptor-like kinase 1 (ALK1, type I receptor) signaling-pathway-related gene expression in the LX-2 cell line, and the inhibitory mechanism of Cpd861 on the activation of LX-2 cells. METHODS: LX-2 cells were treated with TGF beta 1 (5 ng/mL) Cpd861 (0.1 mg/mL), TGF beta 1 (5 ng/mL) plus Cpd861 (5 ng/mL) for 24 h to investigate the effect of Cpd861 on the TGF beta 1/ALK1 pathway. Real-time PCR was performed to examine the expression of alpha-SMA (alpha-smooth muscle actin), ALK1, Id1 (inhibitor of differentiation 1). Western blotting was carried out to measure the levels of alpha-SMA and phosphorylated Smad1, and immunocytochemical analysis for the expression of alpha-SMA. RESULTS: In LX-2 cells, TGF beta 1/ALK1-pathway-related gene expression could be stimulated by TGF beta 1, which led to excessive activation of the cells. Cpd861 decreased the activation of LX-2 cells by reducing the expression of alpha-SMA mRNA and protein expression. This effect was related to inhibition of the above TGF beta 1/ALK1-pathway-related expression of genes such as Id1 and ALK1, and phosphorylation of Smad1 in LX-2 cells, even with TGF beta 1 co-treatment for 24 h. CONCLUSION: Cpd861 can restrain the activation of LX-2 cells by inhibiting the TGF beta 1/ALK1/Smad1 pathway.

Activation of canonical Wnt signaling pathway promotes proliferation and self-renewal of rat hepatic oval cell line WB-F344 in vitro.

AIM: To investigate the effect of activation of canonical Wnt signaling pathway on the proliferation and differentiation of hepatic oval cells in vitro. METHODS: WB-F344 cells were treated with recombinant Wnt3a (20, 40, 80, 160, 200 ng/mL) in serum-free medium for 24 h. Cell proliferation was measured by Brdu incorporation analysis; untreated WB-F344 cells were taken as controls. After treatment with Wnt3a (160 ng/mL) for 24 h, subcellular localization and protein expression of beta-catenin in WB-F344 cells treated and untreated with Wnt3a were examined by immunofluorescence staining and Western blot analysis. CyclinD1 mRNA expression was determined by semi-quantitative reverse-transcript polymerase chain reaction (RT-PCR). The mRNA levels of some phenotypic markers (AFP, CK-19, ALB) and two hepatic nuclear factors (HNF-4, HNF-6) were measured by RT-PCR. Expressions of CK-19 and AFP protein were detected by Western blot analysis. RESULTS: Wnt3a promoted proliferation of WB-F344 cells. Stimulation of WB-F344 cells with recombinant Wnt3a resulted in accumulation of the transcriptional activator beta-catenin, together with its translocation into the nuclei, and up-regulated typical Wnt target gene CyclinD1. After 3 d of Wnt3a treatment in the absence of serum, WB-F344 cells retained their bipotential to express several specific phenotypic markers of hepatocytes and cholangiocytes, such as AFP and CK-19, following activation of the canonical Wnt signaling pathway. CONCLUSION: The canonical Wnt signaling pathway promotes proliferation and self-renewal of rat hepatic oval cells.

Herbal compound 861 regulates mRNA expression of collagen synthesis- and degradation-related genes in human hepatic stellate cells.

AIM: To identify the role of herbal compound 861 (Cpd 861) in the regulation of mRNA expression of collagen synthesis- and degradation-related genes in human hepatic stellate cells (HSCs). METHODS: mRNA levels of collagen types I and III, matrix metalloproteinase 1 (MMP-1), matrix metalloproteinase 2 (MMP-2), membrane type-1 matrix metalloproteinase (MT1-MMP), tissue inhibitor of metalloproteinase 1 (TIMP-1), and transforming growth factor beta1 (TGF-beta1) in cultured-activated HSCs treated with Cpd 861 or interferon-gamma (IFN-gamma) were determined by real-time PCR. RESULTS: Both Cpd 861 and IFN-gamma reduced the mRNA levels of collagen type III, MMP-2 and TGF-beta1. Moreover, Cpd 861 significantly enhanced the MMP-1 mRNA levels while down-regulated the TIMP-1 mRNA expression, increasing the ratio of MMP-1 to TIMP-1 to (6.3 + 0.3)- fold compared to the control group. CONCLUSION: The anti-fibrosis function of Cpd 861 may be mediated by both decreased interstitial collagen sythesis by inhibiting the transcription of collagen type III and TGF-beta1 and increased degradation of these collagens by up-regulating MMP-1 and down-regulating TIMP-1 mRNA levels.

Undifferentiated connective tissue diseases-related hepatic injury.

Hepatic injury is rarely associated with undifferentiated connective tissue diseases (UCTD). We report, here, a case of a middle-aged woman with UCTD-related hepatic injury, including its case history, clinical manifestations, laboratory findings, treatment and its short-term effect. The patient was admitted to the hospital with symptoms of fatigue, anorexia, low-grade fever and skin rashes. She had a past history of left knee joint replacement. Laboratory tests showed elevated levels of serum transaminase, IgG and globulin, accelerated erythrocyte sedimentation rate, eosinophilia and a high titer of antinuclear antibodies (1:320). Imaging studies showed interstitial pneumonitis and hydropericardium. Liver biopsy showed the features which were consistent with those of connective tissue diseases-related polyangitis. After treatment with a low-dose of oral prednisone, both symptoms and laboratory findings were significantly improved. UCTD-related hepatic injury should be considered in the differential diagnosis of connective tissue diseases with abnormal liver function tests. Low-dose prednisone may effectively improve both symptoms and laboratory tests.

Newly proposed fibrosis staging criterion for assessing carbon tetrachloride- and albumin complex-induced liver fibrosis in rodents.

The fibrosis staging criterion (Knodell's) for viral hepatitis in human should not be applied to experimental liver fibrosis in rodents because of differences of species and etiology. Liver fibrosis was induced by carbon tetrachloride (CCl(4)) or albumin antigen-antibody complex in Balb/C mice or Wistar rats. The degree of fibrosis was quantified by staging scores or hydroxyproline measurement or image analysis. Inter- and intra-observer variations of the criterion were also evaluated. Liver fibrosis was divided into seven stages: stage 0, no fibrosis; stage 1, short fibrous tissue in central venule (C); stage 2, fibrous C-C septa appearance; stage 3, C-C fibrous septa developed incompletely; stage 4, C-C septa completely connected (pseudo-lobule); stage 5, C-P (Portal Tract) bridging fibrosis, nodular appearance < or =50%.; and stage 6, nodular appearance >50%. There was significant correlation between the staging scores and hydroxyproline concentration (r = 0.708 in mice, r = 0.841 in rats) and, between staging scores and fibrosis area (r = 0.919 in rats, P < 0.001). The interobserver and intraobserver agreement was consistent (kappa = 0.738 or 0.726, P < 0.001). This staging of hepatic fibrosis in rodents is scientifically rational and repeatable, and is therefore suggested as the criterion for the assessment of experimental hepatic fibrosis in rodent studies.

[Protective effect of compound tongfu granule on intestinal mucosal barrier in patients with cirrhosis of decompensation stage].

OBJECTIVE: To explore the intestinal mucosal barrier protective effect of herbal medicine Compound Tongfu Granule (CTG) in patients with liver cirrhosis of decompensation stage. METHODS: Fifty patients enrolled were randomly assigned to the control group (26 cases) and the CTG group (24 cases), and 30 healthy adults were set up as normal control. After 2-week treatment, the intestinal permeability (IP, represented by urinary lactulose/mannitol excretion rate), plasma endotoxin (EDT) level, and change of enteric bacteria (EB) in patients were observed before and after treatment, and compared with those in the normal control. RESULTS: Before treatment, cirrhotic patients showed significantly higher levels of IP, EDT, and intestinal bacilli, but a lower amount of enteric bifidobacteria as compared with those the normal control. After 2-week treatment, levels of EDT and urinary excretion rate of lactulose in the CTG group were lowered more significantly than those in the control group (P < 0.05), while the amount of bifidobacteria in the CTG group increased accompanied with intestinal bacilli significantly lowered to near the levels in the normal control (P < 0.05, P < 0.01). CONCLUSION: CTG can improve the intestinal barrier function, correct the intestinal bacteria disturbance, and significantly reduce the entero-derived endotoxemia in cirrhotic patients of decompensation stage.

[Relationships between ascitic bacterial DNA and plasma endotoxin, gut flora and intestinal permeability in cirrhotic patients].

OBJECTIVE: To explore the relationship of ascitic bacterial DNA and plasma endotoxin, intestinal permeability, gut flora in cirrhotic patients. METHODS: Fifty-five decompensated cirrhotic patients with ascites were included in the study. A paracentesis was performed for every patient at admission and ascites fluid was collected for bacterial DNA isolation and amplification, plus other routine studies and cultured for aerobic and anaerobic bacteria. Plasma endotoxin, intestinal permeability, and gut flora were studied on the day following admission. Blood from the patients was obtained for routine hematologic, biochemical, and coagulation studies. Thirty healthy subjects were studied as normal controls. RESULTS: No bacteria were found in the ascites cultures in the 55 patients, but bacterial DNA was found in 19 (34.55%). Compared with the bacterial DNA negative group, the bacterial DNA positive group showed a significantly lower level of PTA (t= -3.184, P=0.002), a higher Child-Pugh score (t=3.224, P=0.002) and a higher quantity of WBC in their ascitic fluid (t=4.088, P=0.001). Compared with normal controls, cirrhotic patients showed significantly higher levels of plasma endotoxin (t=13.705, P=0.000), lactulose/mannitol (L/M, t=28.568, P=0.000) in urine, and the quantity of enteric bacilli (t=2.912, P=0.005); the quantity of their intestinal bifidobacteria was significantly lower (t= -3.669, P=0.000). The variables correlative with the presence of bacterial DNA were the quantities of enteric bacilli (P=0.007) and PTA (P=0.011). CONCLUSION: Intestinal bacterial overgrowth plays a key role in the pathogenesis of ascitic bacterial translocation (ABT) in cirrhotic patients. ABT is correlated with the degrees of the liver disease.

[The effect of recombinant interleukin-10/Fc fusion protein on lipopolysaccharide-induced acute lung injury in mice].

OBJECTIVE: To clarify the regulatory role and mechanism of recombinant interleukin-10/Fc (rIL-10/Fc) fusion protein on inflammatory parameters during development of acute lung injury (ALI) induced by lipopolysaccharide (LPS) in a murine model. METHODS: An ALI model was reproduced by intra-tracheal injection of LPS. rIL-10/Fc was administered intraperitoneally. One hundred and thirty-two BALB/c mice were divided into four groups, including saline control group, rIL-10/Fc control group, ALI model group, and rIL-10/Fc treatment group. Twenty-four-hour survival rate was determined in 25 mice of each group. The number of inflammatory cells and inflammatory mediators in bronchial-alveolar lavage fluid (BALF), tumor necrosis factor-alpha (TNF-alpha) and IL-1 beta, and also lung myeloperoxidase (MPO) activity, lung wet/dry (W/D) ratio were determined in the rest of mice. Pathological changes in lung were examined with hematoxylin-eosin (HE) staining, and inflammatory change was evaluated under microscope. RESULTS: Levels of TNF-alpha and IL-1 beta in BALF were substantially increased 4 hours after intra-tracheal LPS (both P<0.01), and they were lowered but without significant difference after rIL-10/Fc administration. However, rIL-10/Fc fusion protein markedly attenuated release of TNF-alpha at 8 hours and 12 hours, and IL-1 beta was lowered at 12 hours after LPS challenge. Pre-treatment with rIL-10/Fc fusion protein significantly improved survival rate at 24 hours in LPS challenged mice (P<0.01). There was no significant difference in cell count in BALF, MPO, lung W/D ratio, after treatment of rIL-10/Fc fusion protein. Obvious inflammatory changes were found in lung was found pathologically at 24 hours after LPS injection, but there was no significant difference compared with ALI mice with rIL-10/Fc fusion protein administration. CONCLUSION: rIL-10/Fc fusion protein inhibits release of TNF-alpha and IL-1 beta in BALF in a LPS-induced ALI murine model. rIL-10/Fc fusion protein improves survival rate in ALI mice by decreasing the release of pro-inflammatory cytokines.

[Scoring system to measure the severity of the multiple organ dysfunction syndrome].

OBJECTIVE: To develop a scoring system to measure the severity of the multiple organ dysfunction syndrome (MODS). METHODS: The clinical data of patients with MODS were collected and analyzed prospectively. Seven indicators were screened out to assess the functions of seven organs. Each indicator was scored from 0 to 4 points, with the 0 point representing the normal organ function, and 1 point to 4 points representing the dysfuntion to failure of the organ. Acording to the index and points a severity scoring system were developed. RESULTS: Seven indicators including the systolic pressure, oxygenation index, conscious state, peripheral blood platelet count, blood total bilirubin, and serum creatinine were used to represent the functions of seven organs. The seven organs included cardiovascular system, lung, brain, coagulative system, liver, kidney, and gastrointestinal tract. The severity scoring system were composed of seven indicators and their points. The total score was 24 points. The mortality increased along with the increase of the points (P<0.001). CONCLUSION: This scoring system can be used to assess the severity of the MODS.