New insights into BMP9 signaling in liver diseases.

Bone morphogenetic protein 9 (BMP9) is a recently discovered cytokine mainly secreted by the liver and is a member of the transforming growth factor ß (TGF-ß) superfamily. In recent years, an increasing number of studies have shown that BMP9 is associated with liver diseases, including nonalcoholic fatty liver disease (NAFLD), liver fibrosis and hepatocellular carcinoma (HCC), and BMP9 signaling may play dual roles in liver diseases. In this review, we mainly summarized and discussed the roles and potential mechanisms of BMP9 signaling in NAFLD, liver fibrosis and HCC. Specifically, this article will provide a better understanding of BMP9 signaling and new clues for the treatment of liver diseases.

Efficacy and safety of tauroursodeoxycholic acid in the treatment of liver cirrhosis: a double-blind randomized controlled trial.

No direct comparison of tauroursodeoxycholic acid (TUDCA) and ursodeoxycholic acid (UDCA) has yet been carried out in the treatment of liver cirrhosis in China. We designed a double-blind randomized trial to evaluate the potential therapeutic efficacy of TUDCA in liver cirrhosis, using UDCA as parallel control. The enrolled 23 patients with liver cirrhosis were randomly divided into TUDCA group (n=12) and UDCA group (n=11), and given TUDCA and UDCA respectively at the daily dose of 750 mg, in a randomly assigned sequence for a 6-month period. Clinical, biochemical and histological features, and liver ultrasonographic findings were evaluated before and after the study. According to the inclusion criteria, 18 patients were included in the final analysis, including 9 cases in both two groups. Serum ALT, AST and ALP levels in TUDCA group and AST levels in UDCA group were significantly reduced as compared with baseline (P<0.05). Serum albumin levels were significantly increased in both TUDCA and UDCA groups (P<0.05). Serum markers for liver fibrosis were slightly decreased with the difference being not significant in either group. Only one patient in TUDCA group had significantly histological relief. Both treatments were well tolerated and no patient complained of side effects. It is suggested that TUDCA therapy is safe and appears to be more effective than UDCA in the treatment of liver cirrhosis, particularly in the improvement of the biochemical expression. However, both drugs exert no effect on the serum markers for liver fibrosis during 6-month treatment.

[Effects of cAMP-response element binding protein-1 (CREB-1) on transforming growth factor-b3 (TGFb3) mRNA expression and promoter activity in hepatic stellate cells].

OBJECTIVE: To investigate the effects of cAMP-response element binding protein-1 (CREB-1) on transforming growth factor-b3 (TGF b3) mRNA expression and promoter activity in hepatic stellate cells (HSCs). METHODS: Freshly isolated HSCs from rats were divided into six groups: CREB-1 expression plasmid transfected group (C), siRNA-CREB-1 plasmid transfected group (S), negative control group (N), forskolin treated group (F), H-89 treated group (H), and blank group (B). Rats in each group were further sub-divided according to whether (+) or not (-) they were exposed to exogenous TGF b3. TGF b3 mRNA expression was measured by real time quantitative PCR. HSCs of the C, S, N, F, H and B groups were transfected with the TGF b3 promoter luciferase reporter plasmid (PGL3-TGF b3-P; W group), the TGF b3 promoter luciferase reporter plasmid with CRE mutation (PGL3-basic-TGF b3P-mCRE; M group) and the renilla luciferase control plasmid (pRL-SV40; control group). TGF b3 promoter activity was assessed by luciferase reporter assays. RESULTS: Compared to N(-), the TGF b3 mRNA expression was reduced to 0.69+/-0.15 in S(-) (P less than 0.05) and increased to 4.68+/-2.76 in C(-) (P more than 0.05). Compared to B(-), the TGF b3 mRNA expression was reduced to 0.57+/-0.08 in H(-) (P less than 0.05). The differences between N(+) and N(-), S(+) and S(-), B(+) and B(-), and H(+) and H(-) were all significant (P less than 0.05). The values of TGF b3 promoter activity in S(W), N(W), and C(W) were 0.062+/-0.013, 0.122+/-0.011, and 0.165+/-0.016 (P less than 0.05), but the changes of TGF b3 promoter activity in S(M), N(M), and C(M) were not significant (P more than 0.05). The values of TGF b3 promoter activity in H(W), B(W), and F(W) were 0.154+/-0.010, 0.188+/-0.016, and 0.276+/-0.031 (P less than 0.05), but the changes of TGF b3 promoter activity in H(M), B(M), and F(M) were not significant (P more than 0.05). CONCLUSION: Increased levels of CREB-1 mRNA or p-CREB-1 up-regulate the TGF b3 mRNA expression and promoter activity in rat HSCs. The CRE site in the TGF b3 promoter is critical for this effect, and the gene's activity becomes significantly decreased when the site is missing. Exogenous TGF b3 enhances expression of endogenous TGF b3 in rat HSCs.

Difference between type 2 gastroesophageal varices and isolated fundic varices in clinical profiles and portosystemic collaterals.

BACKGROUND: There is significant heterogeneity between gastroesophageal varices (GOV2) and isolated gastric varices (IGV1). The data on the difference between GOV2 and IGV1 are limited. AIM: To determine the etiology, clinical profiles, endoscopic findings, imaging signs, portosystemic collaterals in patients with GOV2 and IGV1. METHODS: Medical records of 252 patients with gastric fundal varices were retrospectively collected, and computed tomography images were analyzed. RESULTS: Significant differences in routine blood examination, Child-Pugh classification and MELD scores were found between GOV2 and IGV1. The incidence of peptic ulcers in patients with IGV1 (26.55%) was higher than that of GOV2 (11.01%), while portal hypertensive gastropathy was more commonly found in patients with GOV2 (22.02%) than in those with IGV1 (3.54%). Typical radiological signs of cirrhotic liver were more commonly observed in patients with GOV2 than in those with IGV1. In patients with GOV2, the main afferent vessels were via the left gastric vein (LGV) (97.94%) and short gastric vein (SGV) (39.18%). In patients with IGV1, the main afferent vessels were via the LGV (75.61%), SGV (63.41%) and posterior gastric vein (PGV) (43.90%). In IGV1 patients with pancreatic diseases, spleno-gastromental-superior mesenteric shunt (48.15%) was a major collateral vessel. In patients with fundic varices, the sizes of gastric/esophageal varices were positively correlated with afferent vessels (LGVs and PGVs) and efferent vessels (gastrorenal shunts). The size of the esophageal varices was negatively correlated with gastrorenal shunts in GOV2 patients. CONCLUSION: Significant heterogeneity in the etiology and vascular changes between GOV2 and IGV1 is useful in making therapeutic decisions.

Effects of p-CREB-1 on transforming growth factor-ß3 auto-regulation in hepatic stellate cells.

Previous studies have demonstrated that transforming growth factor-ß3 (TGF-ß3) protected liver against fibrosis in vivo and vitro, but its regulation is poorly understood. In addition, the cAMP-responsive element (CRE) in TGF-ß3 promoter is recognized as an important regulatory site for TGF-ß3 auto-regulation. Thus, we hypothesize that transcription factor CRE-binding protein-1 (CREB-1) regulates the auto-induction of TGF-ß3 in hepatic stellate cells (HSCs). We used exogenous TGF-ß3 to activate the signal pathway of TGF-ß3 auto-regulation in HSCs, results indicated that exogenous TGF-ß3 could up-regulate the protein and mRNA expressions of TGF-ß3, and provoke the phosphorylation of CREB-1 on Ser-133, besides, it could induce the DNA binding activity of p-CREB-1 and activate TGF-ß3 promoter as well. Additionally, we used pGenesil-1.1-shRNA-CREB-1 and pRSV-CREB-1 expression vector to silence and up-regulate CREB-1 gene expression respectively, and the results indicated that inhibition of CREB-1 suppressed exogenous TGF-ß3 stimulation of TGF-ß3 mRNA and protein expressions in HSCs, whereas up-regulation of CREB-1 induced this stimulation. Our results indicate that exogenous TGF-ß3 up-regulates the activity of TGF-ß3 promoter by activating CREB-1, then induces the mRNA and protein expressions of TGF-ß3. Especially, p-CREB-1 is a critical transcription factor in mediating TGF-ß3 auto-induction.

[Effects of exogenous transforming growth factor-ß3 on the activities of its promoter and cAMP-responsive element binding protein-1 in rat hepatic stellate cell].

OBJECTIVE: To explore the effects of exogenous transforming growth factor-ß3 (TGF-ß3) on the activities of its promoter and cAMP-responsive element binding protein-1 (CREB-1) in rat hepatic stellate cell (HSC-T6). METHODS: HSC-T6 was cultured and treated with or without exogenous TGF-ß3 (10 µg/L). Then cell extracts, total RNA and nuclear proteins were collected at different time points. The specimens were detected by luciferase reporter assay, Western blotting and real-time RT-PCR (reverse transcription-polymerase chain reaction) respectively. RESULTS: After treatment, the activity of TGF-ß3 promoter peaked at 24 h (10.68 ± 0.57 vs 4.83 ± 0.56, 2.2 folds vs control). And the mutational CRE site completely blocked the activity of TGF-ß3 promoter (0.73 ± 0.03, P < 0.05). In addition, exogenous TGF-ß3 increased the expression of phospho-CREB-1 in a time-dependent manner. It peaked at 1 h (2.0 folds vs control) and declined slowly. And exogenous TGF-ß3 had no effect on the mRNA and protein expressions of CREB-1 (P > 0.05). CONCLUSION: The activity of TGF-ß3 promoter is up-regulated by exogenous TGF-ß3. And CRE site in TGF-ß3 promoter region is important for the transcription of TGF-ß3 gene in HSC-T6. While activating CREB-1, exogenous TGF-ß3 has no effect on the expressions of CREB-1 protein and mRNA.

[Effects of exogenous TGF-ß3 on the expression of endogenous TGF-ß3 in hepatic stellate cell-T6 (HSC-T6)].

OBJECTIVE: To investigate the effects of exogenous TGF-ß3 on the expression of endogenous TGF-b3 in hepatic stellate cell (HSC). METHODS: HSCs were cultured and divided into two groups: TGF-ß3 group and blank control group, the cells of TGF-ß3 group were exposed to TGF-b3 (10 ng/ml), whereas the blank control group was not treated. The cells were incubated in the presence of exogenous TGF-ß3 and then (1) were harvested at 0h, 1h, 2h, 4h, 12h, 24h, and real time PCR was performed to detect the mRNA expression of endogenous TGF-ß3. (2) The cells were collected at 0h, 1h, 6h, 12h, and western-blot was used to detect the protein synthesis of endogenous TGF-ß3 in HSC; (3) The cell culture supernatant was harvested at 0h, 1h, 2h, 4h, 8h, 14h, 24h, and ELISA was performed to measure the total protein of extracellular TGF-ß3; HSCs were treated with TGF-ß3 (10 ng/ml) for 2h. The cells were then incubated in serum-free medium and the cell culture supernatant was harvested at 2.25h, 2.5h, 3h, 4h, 6h, 10h and 14h. ELISA was used to detect the extracellular secret ion of endogenous TGF-ß3 by HSCs. RESULTS: (1) Exogenous TGF-ß3 treatment induced a marked increase in TGF-ß3 mRNA expression. By 2h of exogenous TGF-ß3 treatment, maximal TGF-ß3 mRNA expression levels (2.796 ± 0.518) of 2.74 fold above control values (1.022 ± 0.038) was reached (P < 0.05). Thereafter, TGF-ß3 mRNA expression level declined, and the expression level was maintained at level of 1.45-fold for at least 10h and was 1.18-fold above control values by 24h TGF-ß3 treatment (P < 0.05); (2) No significant difference about the intracellular protein expression level of endogenous TGF-ß3 was found between two groups. (P > 0.05); (3) The total expression level of TGF-ß3 reached a peak [(18.931 ± 2.904) ng/ml] at 4h after TGF-ß3 treatment (1.89-fold higher than basic TGF-ß3 (10 ng/ml). After that, it slowly declined. The expression peak [(0.835 ± 0.027) ng/ml] induction of extracellular secreted TGF-ß3 was at 3h (32.12-fold higher than control [(0.026 ± 0.022) ng/ml], (P < 0.05). Thereafter, TGF-ß3 slowly decreased after the peak time, and their expressions were still statistically significant as compared to the control (P < 0.05). CONCLUSION: Exogenous TGF-ß3 could increase the expression of endogenous TGF-ß3 mRNA and extracellular secreted TGF-ß3 protein obviously.

Zhikang Capsule Ameliorates Inflammation, Drives Polarization to M2 Macrophages, and Inhibits Apoptosis in Lipopolysaccharide-induced RAW264.7 Cells.

OBJECTIVE: To explore the anti-inflammatory effect of the traditional Chinese medicine Zhikang capsule (ZKC) on lipopolysaccharide (LPS)-induced RAW264.7 cells. METHODS: Safe concentrations of ZKC (0.175, 0.35, and 0.7 mg/mL) were used after the half-maximal inhibitory concentration (IC50) of RAW264.7 cells was calculated through the CCK-8 assay. In addition, the optimal intervention duration of ZKC (0.7 mg/mL) on RAW264.7 cells was determined to be 6 h, since all proinflammatory mediators [tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1ß), inteleukin-6 (IL-6), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and monocyte chemotactic protein-1 (MCP-1)] had a decreasing tendency and relatively down-regulated mRNA expression levels as compared with other durations (4, 8, and 12 h). RAW264.7 cells were pretreated with ZKC at various concentrations (0.175, 0.35 and 0.7 mg/mL) for 6 h and then stimulated with LPS (1 µg/mL) for an additional 12 h. RESULTS: In terms of inflammation, ZKC could reverse LPS-induced upregulation of TNF-α, IL-1ß, IL-6, COX-2, iNOS, and MCP-1 at both the mRNA and protein levels in RAW264.7 cells in a dose-dependent manner. In terms of the NF-κB signaling pathway, ZKC could reduce phosphorylated p65 and promote M2 polarization of RAW264.7 cells under LPS stimulation in a dose-dependent manner. Moreover, ZKC exhibited a protective effect on macrophages from apoptosis. CONCLUSION: ZKC exhibited obvious antiinflammatory and anti-apoptotic effects on LPS-induced RAW264.7 cells at the cellular level, and a weakened NF-κB signaling pathway may be a potential significant target.

[Effects of transforming growth factor beta 3 on the histopathology and expression of collagen I in experimental hepatic fibrotic rats].

OBJECTIVE: To investgate the effects of TGFbeta3 on rat hepatic fibrosis. METHODS: The TGFbeta3 cDNA was cloned into rAAV2 vector. Rats were randomly divided into four groups: normal control group, model group, negative control group and TGFbeta3 group. Hepatic fibrosis was induced by hypodermic injection of 40% CCl(4). Recombinant AAV2-TGFbeta3 viral particles were injected via the vena caudalis one week before CCl(4) treatment. Rats were executed 8 weeks after CCl(4) treatment, global histological change was observed after HE staining, the distribution of the collagen fibers was observed after masson staining, histochemistry was done to observe the expression of collagen I; The positive area rate of the collagen fibers and the average optical rate of collagen I were quantified. RESULTS: HE staining indicated that collagen fibers were reduced in the TGFbeta3 group. Masson staining shown that the collagen fibers were distributed around the blood vessel, in the portal area and disse space. Compared to the model group (13.2%+/-2.2%) and negative control group (12.3%+/-1.5%), the collagen fibers in liver tissues of TGFbeta3 group (7.7%+/-1.5%) were significantly decreased (q = 9.456, P < 0.01; q = 8.217, P < 0.01). Histochemistry indicated that the collagen fibers of liver tissues of TGFbeta3 group (0.185+/-0.033) were significantly higher than those in the model group (0.252+/-0.042) and the negative control group (0.230+/-0.029), (q = 6.228, P < 0.01; q = 4.346, P < 0.01). CONCLUSION: TGFbeta3 alleviates the damage to hepatic cell and the level of fibrosis in CCl(4) treated rats and inhibits the expression of collagen I.

[Influence of recombinant transforming growth factor-beta3 on collagen synthesis and deposition: experiment with rat cell model of liver fibrosis].

OBJECTIVE: To investigate the influence of recombinant transforming growth factor-beta3 (TGF-beta3) on collagen synthesis and deposition. METHODS: Plasmids pcDNA3.1 (+)-TGF-beta3 and pcDNA3.1 (+)-TGF-beta1 were constructed. Rat hepatic stellate cells (HSCs) of the strain HSC-T6 were cultured as cell model of fibrosis and divided into 4 groups: blank control group, pcDNA3.1-enhanced green fluorescent protein (EGFP)-transfected group (negative control group), pcDNA3.1 (+)-TGF-beta1 transfected group, and pcDNA3.1 (+)-TGF-beta3 transfected group. A positive cell clone stably and highly expressing TGF-beta1 was established after being screened by G418 medium. pcDNA3.1 (+)-TGF-beta3 was transfected into the positive clone. Real-time PCR was used to detect the mRNA expression of TGF-beta3. Western blotting was used to detect the protein expression of TGF-beta1, collagen I, matrix metalloproteinase (MMP)-2, MMP-9, and tissue inhibitor of metallo-proteinase (TIMP)-1. RESULTS: There was no significant difference in the TGF-beta1 mRNA expression between the TGF-beta1 positive clone group and the TGF-beta3 transfected group. The mRNA and protein expression levels of TGF-beta1, collagen I, MMP-2, and TIMP-1 of the TGF-beta1 positive clone group were all significantly higher than those of the blank control, negative control groups (all P < 0.05), and the MMP-9 mRNA expression of the TGF-beta1 positive clone group was significantly lower than those of the blank control and negative control groups (all P < 0.05); and the mRNA and protein expression levels of MMP-9 of the TGF-beta1 positive clone group were significantly lower than those of the blank control and negative control groups (all P < 0.05). The TGF-beta1 and MMP-2 mRNA expression levels of the TGF-beta3 transfected group were not significantly different from those of the positive clone group (all P > 0.05), the mRNA expression levels of collagen I and TIMP-1 of the TGF-beta3 transfected group were significantly lower than those of the positive clone group (both P < 0.05), and the mRNA expression level of MMP-9 of the TGF-beta3 transfected group was significantly higher than that of the positive clone group (P < 0.05). The protein expression levels of TGF-beta1, collagen I, and TIMP-1 of the TGF-beta3 transfected group were all significantly lower than those of the positive clone group (P < 0.05), the protein expression level of MMP-9 of the TGF-beta3 transfected group was significantly higher than that of the positive clone group (P < 0.05), and the protein expression level of MMP-2 of the TGF-beta3 transfected group was not significantly different from that of the positive clone group. CONCLUSION: Recombinant TGF-beta3 eukaryotic expression vector reduces the synthesis of collagen and inhibits the collagen deposition by adjusting the expression of matrix metalloproteinases and their inhibitors.

[Effects of transforming growth factor-beta 3 gene transfer on type I collagen synthesis of hepatic stellate cells].

OBJECTIVE: To observe the effects of transforming growth factor-beta 3 gene transfer on type I collagen synthesis of cells of HSC-T6. METHODS: Transforming growth factor-beta 1 expression plasmid and transforming growth factor-beta 3 expression plasmid were constructed. The recombinant expression plasmids pcDNA3.1(+)-TGF beta 1 and pcDNA3.1(+)-TGF beta 3 were respectively transfected and cotransfected into cultured HSC-T6 cells; expression of TGF betav1, TGF beta 3, type I collagen mRNA were detected by real-time quantitative PCR, expression of TGF beta 1 and type I collagen protein were detected by Western blot. The recombinant expression plasmid pcDNA3.1(+)-TGF beta 1 was transfected into cultured HSC-T6 cells; positive clones were selected by G418. The positive clones were transfected by the recombinant expression plasmid pcDNA3.1(+)-TGF beta 3; expression of TGF beta 1, TGF beta 3 and type I collagen mRNA were detected by real-time quantitative PCR; expression of TGF beta 1 and type I collagen protein were detected by Western blot. RESULTS: HSC-T6 was transfected by recombinant expression plasmid pcDNA3.1(+)-TGF beta 1 and pcDNA3.1(+)-TGF beta 3 and the transfection efficiency was 28.2%. After the cells were transfected with pcDNA3.1-TGF beta 3, type I collagen mRNA and the protein expression in the cells were higher than those in the untransfected cells (control group) (P < 0.05). The increase reached to the maximal at 72 h after the transfection. Expressions of type I collagen mRNA and the protein in the cells transfected by pcDNA3.1(+)-TGF beta 1 were higher than in those cotransfected by pcDNA3.1(+)-TGF beta 1 and pcDNA3.1(+)-TGF beta 3 (P < 0.05). TGF beta 1 protein, type I collagen mRNA and type I collagen protein expression significantly decreased in the clones transfected by recombinant expression plasmid pcDNA3.1(+)-TGF beta 3 (P < 0.05), but the changes of TGF beta 1 mRNA were not significant (P > 0.05). CONCLUSIONS: Expression of type I collagen increased after the cultured HSC-T6 cells were transfected by TGF beta 3 gene. The significant decrease of the expression of type I collagen of the TGF beta 3 gene transfected positive clones suggests that TGF beta 3 could inhibit the occurrence of liver fibrosis.

Unusual cause of lesions in the descending duodenum and liver: A case report and review of literature.

The descending duodenum is rarely involved in Schistosoma japonicum (S. japonicum) infection. Here, we report a case of acute Schistosoma infection, which presented with abdominal pain, abdominal distension and irregular fever. Tumor-like lesions were observed in the descending duodenum. Simultaneously, heterogeneity in hepatic perfusion was demonstrated by dynamic computed tomography scanning. Biopsy of the descending duodenum showed the deposition of Schistosoma eggs. Following administration of the antihelminthic drug praziquantel, the patient showed rapid clinical improvement. In conclusion, we report a patient with acute S. japonicum infection presenting as tumor-like lesions in the descending duodenum and heterogeneity of blood perfusion in liver parenchyma.

Changes in hydrogen sulfide in rats with hepatic cirrhosis in different stages.

This study aimed to observe changes in the hydrogen sulfide (H2S) system in the blood and liver tissue of rats with hepatic cirrhosis at different stages by studying the effect of H2S on the course of hyperdynamic circulation in rats with hepatic cirrhosis. H2S concentration in the blood from the portal vein and inferior vena cava of hepatic cirrhosis rat model induced with carbon tetrachloride was detected on the 15th, 30th, and 52nd day. The expression of cystathionine ß-synthase (CBS) and cystathionine γ-lyase (CSE) protein, and CBS and CSE mRNA in the liver was detected by immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR), respectively. The results indicated that H2S concentration in the blood from the portal vein and inferior vena cava of rats with hepatic cirrhosis was significantly lower than that in the control group. H2S was gradually decreased with the development of the disease and significantly lower in the blood from portal vein than in the blood of inferior vena cava at the mid-stage and the late stage groups. The expression levels of CBS and CSE protein, and CBS and CSE mRNA in the livers with hepatic cirrhosis at different stages were all higher than those in the control group, and the expression gradually increased with the development of the disease. The expression of CBS was lower than CSE in the same stages. The results indicated that the CSE mRNA was expressed predominantly in the cirrhosis groups as compared with CBS mRNA. Among experimental rats, the H2S system has an important effect on the occurrence and development of hyperdynamic circulation in rats with hepatic cirrhosis. This finding adds to the literature by demonstrating that H2S protects vascular remodelling in the liver, and that CSE is indispensable in this process.

[Expression of netrin-1 in placenta from patients with pre-eclampsia and the relation to placental angiogenesis].

OBJECTIVE: To investigate the expression of netrin-1 in placenta from patients with pre-eclampsia and the relation to placental angiogenesis. METHODS: Twenty patients with pre-eclampsia (12 mild cases and 8 severe cases) and 20 normal late pregnant women were investigated. The expression of netrin-1 mRNA and protein was determined with RT-PCR and Western blotting respectively. The placenta vascular density was examined by immunohistochemical F8 staining. RESULTS: (1) The values of netrin-1 mRNA in normal group and pre-eclampsia groups were 0.51 +/- 0.08 and 0.41 +/- 0.06; The values of netrin-1 protein in normal group and pre-eclampsia groups were 26.4 +/- 1.8 and 20.5 +/- 1.3 (P < 0.01). The values of netrin-1 mRNA in mild and severe pre-eclampsia groups were 0.48 +/- 0.08 and 0.34 +/- 0.07; The values of netrin-1 protein in mild and severe pre-eclampsia groups were 22.8 +/- 1.3 and 18.2 +/- 1.0 (P < 0.01). (2) The placenta vascular density in pre-eclampsia (54 +/- 8) was significantly reduced than in normal group (65 +/- 10) (P < 0.01); the decrease was even more obvious in severe group (48 +/- 7) than in mild group (60 +/- 9) (P < 0.01). (3) There was positive correlation between the expression of netrin-1 mRNA and protein and placenta vascular density in normal pregnancy (r = 0.67 and 0.71, P < 0.01), as well as in pre-eclampsia group (r = 0.61 and 0.70, P < 0.01), in mild pre-eclampsia group (r = 0.69 and 0.73, P < 0.01), and in severe pre-eclampsia group (r = 0.71 and 0.75, P < 0.01). CONCLUSION: The study shows that the decrease of netrin-1 in placenta of patients with pre-eclampsia is one of the possible reasons for the reduction of placenta vascular density.

[Changes of mast cells and protease activated receptor-2 in experimental rat liver fibrosis].

OBJECTIVE: To explore the quantity of mast cells and the role of protease activated receptor-2 (PAR-2) in experimental rat liver fibrosis. METHODS: Rats were sacrificed at 0, 2, 4, 8, and 12 weeks after subcutaneous injection of CCl(4). Mast cells were displayed by toluidine blue stain. The content of liver hydroxyproline was measured by the method of base hydrolyzate. The mRNA expression and the protein expression of PAR-2 in livers were detected by RT-PCR and immunohistochemistry at each time point. RESULTS: In normal rat livers there were a few mast cells (2.5+/-1.0) distributed along the hepatic portal areas. In the cirrhosis model group the number of mast cells in the livers increased degree by degree (2 weeks vs 4 weeks vs 8 weeks, 9.1+/-0.5 vs 15.7+/-3.0 vs 32.0+/-3.3; P less than 0.05), and they were distributed densely around the hepatic portal areas and the central veins. The content of liver hydroxyproline increased progressively from 0 to 12 weeks. In normal livers PAR-2 mRNA was hardly detected, at 2 weeks there was some expression of PAR-2 mRNA (PAR-2/beta-actin 0.15+/-0.01, P less than 0.05), at 4 weeks its expression increased (PAR-2/beta-actin 0.35+/-0.02, P less than 0.05) and it maintained a higher level (PAR-2/beta-actin 0.80+/-0.02, P less than 0.05) since then. The changing trend of the protein expression of PAR-2 was the same as that of PAR-2 mRNA expression. CONCLUSIONS: PAR-2 mRNA expression and the protein expression of PAR-2 were consistent with the increase of the mast cells, and the content of liver hydroxyproline may play an important role in mediating liver fibrosis.

Primary biliary cirrhosis-autoimmune hepatitis overlap syndrome: simplified criteria may be effective in the diagnosis in Chinese patients.

OBJECTIVE: To evaluate the Paris criteria, the revised diagnostic criteria and the simplified diagnostic scoring system in the diagnosis of primary biliary cirrhosis (PBC)-autoimmune hepatitis (AIH) overlap syndrome in Chinese patients. METHODS: Medical records of the patients who were diagnosed with PBC at the Union Hospital and Tongji Hospital, Tongji Medical University, Huazhong University of Science and Technology (Wuhan, Hubei Province, China) from 2003 to 2012 were retrospectively reviewed. The overlap syndrome was diagnosed based on the Paris criteria, the revised criteria and the simplified criteria, respectively. Patients' clinical characteristics, laboratory examination results and histological findings were collected. The sensitivity and specificity of the three criteria for diagnosing PBC-AIH overlap syndrome were calculated. RESULTS: PBC-AIH overlap syndrome was diagnosed in 2, 13 and 10 patients with PBC based on the Paris, the revised and the simplified criteria, respectively. The sensitivity and specificity of the simplified criteria in diagnosing the overlap syndrome was 90.0% and 98.2%, which were the highest among the three criteria, followed by the revised criteria. The Paris criteria showed a high specificity (100%) but a relatively low sensitivity (20.0%). In addition, some patients who did not fulfil the Paris criteria still benefited from the immunosuppressive agents. CONCLUSIONS: For Chinese patients with the PBC-AIH overlap syndrome, the simplified criteria appear to be the most efficacious compared with the Paris criteria and the revised criteria. Further studies should be performed to confirm these observations with respect to long-term outcomes and therapeutic implications.