HgS and Zuotai differ from HgCl2 and methyl mercury in intestinal Hg absorption, transporter expression and gut microbiome in mice.

Mercury (Hg) is generally considered as a toxic metal; yet the biological outcomes of Hg-containing compounds are highly dependent upon their chemical forms. We hypothesize that mercury sulfide (HgS) is different from HgCl2 and methylmercury (MeHg) in producing intestinal Hg absorption and disruption of gut microbiome. To test this hypothesis, mice were given orally with HgS (α-HgS, 30 mg/kg), Zuotai (ß-HgS, 30 mg/kg), HgCl2 (33.6 mg/kg, equivalent Hg as HgS), or MeHg (3.1 mg/kg, 1/10 Hg as HgS) for 7 days. Accumulation of Hg in the duodenum and ileum after HgCl2 (30-40 fold) and MeHg (10-15 fold) was higher than HgS and Zuotai (~2-fold). HgCl2 and MeHg decreased intestinal intake peptide transporter-1 and Ost-ß, and increased ileal bile acid binding protein and equilibrative nucleoside transporter-1. The efflux transporters ATP-binding cassette sub-family C member-4 (Abcc4), Abcg2, Abcg5/8, and Abcb1b were increased by HgCl2 and to a lesser extent by MeHg, while HgS and Zuotai had minimal effects. Bacterial DNA was extracted and subjected to 16S rDNA sequencing. Operational taxonomic unit (OTU) results showed that among the 10 phyla, HgS increased Firmicutes, Proteobacteria, while HgCl2 increased Bacteroidetes, Cyanobacteria and decreased Firmicutes; among the 79 families, HgS increased Rikenellaceae, Lactobacillaceae, Helicobacteraceae, and decreased Prevotellaceae, while HgCl2 increased Odoribacteraceae, Porphyromonadaceae, and decreased Lactobacillaceae; among the 232 genus/species, HgS and Zuotai affected gut microbiome quite differently from HgCl2 and MeHg. qPCR analysis with 16S rRNA confirmed sequencing results. Thus, chemical forms of mercury are a major determinant for intestinal Hg accumulation, alterations in transporters and disruption of microbiome.

[The Effect of p-JAK1/p-STAT3 Signaling Way in Intervening NAFLD by Blueberry Probiotic Serum].

OBJECTIVES: To explore the mechanism of interlukin-22 (IL-22)-mediated phosphor-Janus kinase-1(p-JAK1)/phosphor-signal transducer and activator of transcription 3 (p-STAT3) signaling way in the experiment of improving non-alcholic fatty liver disease (NAFLD) by blueberry probiotic serum. METHODS: The rat serums with low-, medium-, and high-dose of 10% blueberry probiotics, as well as saline were prepared. NAFLD model was built by inducing normal liver cell line L-02 with free fatty acid (FFA).NAFLD model cells were cultured with saline serum (model group), low-, medium-, and high-dose blueberry probiotics serums (low-, medium-, and high-dose serum groups) , respectively .Normal liver cell group (normal group) was cultured with saline serum . Oil Red O staining was used to detect the lipid deposition in the cells; the intracellular level of triglyceride (TG) was quantitatively determined; the gene and protein expressions of IL-22, p-JAK1, p-STAT3, sterol-regulatory element binding protein-1c (SREBP-1c ) were detected by RT-PCR, Western blot and immunofluorescence methods. RESULTS: Twenty-four hours after modeling, a large amount of lipid deposition could be observed in model group. Compared with normal group, model group showed lower gene and protein expression levels of IL-22, p-JAK1 and p-STAT3 (P <0.01), and higher SREBP-1c and TG levels (P <0.01).Compared with model group, TG level and the lipid deposition in low-, medium-, and high-dose blueberry probiotics serum groups were gradually reduced. High-dose serum group showed higher gene and protein expression levels of IL-22, p-JAK1, p-STAT3 and lower SREBP-1c compared with the model, low-, and medium-dose serum groups (P <0.01). No significant [CM(155.3mm]differences in gene and protein levels between low- andmedium-doseserum groups were found (P >0.05). CONCLUSION: The blueberry probiotics could antagonize the NAFLD via p-JAK1/p-STAT3 signaling way.

Liver expression of Nrf2-related genes in different liver diseases.

BACKGROUND: The KEAP1-Nrf2 antioxidant signaling pathway is important in protecting liver from various insults. However, little is known about the expression of Nrf2-related genes in human liver in different diseases. METHODS: This study utilized normal donor liver tissues (n=35), samples from patients with hepatocellular carcinoma (HCC, n=24), HBV-related cirrhosis (n=27), alcoholic cirrhosis (n=5) and end-stage liver disease (n=13). All of the liver tissues were from the Oriental Liver Transplant Center, Beijing, China. The expressions of Nrf2 and Nrf2-related genes, including its negative regulator Kelch-like ECH-associated protein 1 (KEAP1), its targeted gene NAD(P)H-quinone oxidoreductase 1 (NQO1), glutamate-cysteine ligase catalytic subunit (GCLC) and modified subunit (GCLM), heme oxygenase 1 (HO-1) and peroxiredoxin-1 (PRDX1) were evaluated. RESULTS: The expression of Nrf2 was decreased in HCC, increased in alcoholic cirrhosis and end-stage liver disease. The expression of KEAP1 was increased in all of the liver samples. The most notable finding was the increased expression of NQO1 in HCC (18-fold), alcoholic cirrhosis (6-fold), end-stage liver disease (5-fold) and HBV-related cirrhosis (3-fold). Peri-HCC also had 4-fold higher NQO1 mRNA as compared to the normal livers. GCLC mRNA levels were lower only in HCC, as compared to the normal livers and peri-HCC tissues. GCLM mRNA levels were higher in HBV-related cirrhosis and end-stage liver disease. HO-1 mRNA levels were increased in all liver tissues except for HCC. Peri-HCC had higher PRDX1 mRNA levels compared with HCC and normal livers. CONCLUSION: Nrf2 and Nrf2-related genes are aberrantly expressed in the liver in different diseases and the increase of NQO1 was the most notable finding, especially in HCC.

[Gene expression changes of regucalcin and prohibitin in cirrhotic rat liver and the related effects of compound glutathione inosine injection intervention].

OBJECTIVE: To study expression of regucalcin (RGN) and prohibitin (PHB) genes in cirrhotic rat liver and to investigate the related effects of compound glutathione inosine injection (CGII) intervention. METHODS: Forty male Wistar rats were randomly divided into a control group (n=12) and a model group (n=28).The model was established by injecting sterile porcine serum (0.5 mL) into the rat abdominal cavity, twice weekly for 8 consecutive weeks; the control group rats were treated with physiological saline injection (0.5 mL) into the abdominal cavity with the same frequency and time span. During the modeling period, four rats from the model group were randomly selected at different time points to examine changes in liver pathology. Upon pathology confirmation of liver cirrhosis, the porcine serum injection was terminated. The remaining 24 rats in the model group were randomly divided into a fibrosis group and a CGII treatment group.The CGII group received CGII (intramuscular injection of 0.018 mL 100g-1 body weight) once a day for 6 continuous weeks; the fibrosis rats were treated with the same dosage of physiological saline with the same frequency and time span.Liver tissue morphology was examined by both hematoxylin-eosin and Masson's staining. RGN and PHB expression at the mRNA and protein levels in liver tissues were detected by real time RT-PCR and immunohistochemical staining, respectively. RESULTS: Both the mRNA and protein expression levels of RGN and PHB were significantly lower in the liver tissues of the fibrosis group than in the control group.CGII intervention led to significant alleviation of the liver fibrosis severity; moreover, the mRNA and protein expression levels of RGN and PHB were significantly higher than those in the fibrosis group. CONCLUSION: Down-regulation of regucalcin and prohibitin gene expression might contribute to the pathogenesis of liver cirrhosis.

Autoimmune hepatitis-primary biliary cholangitis overlap syndrome complicated by various autoimmune diseases: A case report.

BACKGROUND: Autoimmune hepatitis (AIH) and primary biliary cholangitis (PBC) are two common clinical autoimmune liver diseases, and some patients have both diseases; this feature is called AIH-PBC overlap syndrome. Autoimmune thyroid disease (AITD) is the most frequently overlapping extrahepatic autoimmune disease. Immunoglobulin (IgG) 4-related disease is an autoimmune disease recognized in recent years, characterized by elevated serum IgG4 levels and infiltration of IgG4-positive plasma cells in tissues. CASE SUMMARY: A 68-year-old female patient was admitted with a history of right upper quadrant pain, anorexia, and jaundice on physical examination. Laboratory examination revealed elevated liver enzymes, multiple positive autoantibodies associated with liver and thyroid disease, and imaging and biopsy suggestive of pancreatitis, hepatitis, and PBC. A diagnosis was made of a rare and complex overlap syndrome of AIH, PBC, AITD, and IgG4-related disease. Laboratory features improved on treatment with ursodeoxycholic acid, methylprednisolone, and azathioprine. CONCLUSION: This case highlights the importance of screening patients with autoimmune diseases for related conditions.

Silent information regulator sirtuin 1 ameliorates acute liver failure via the p53/glutathione peroxidase 4/gasdermin D axis.

BACKGROUND: Acute liver failure (ALF) has a high mortality with widespread hepatocyte death involving ferroptosis and pyroptosis. The silent information regulator sirtuin 1 (SIRT1)-mediated deacetylation affects multiple biological processes, including cellular senescence, apoptosis, sugar and lipid metabolism, oxidative stress, and inflammation. AIM: To investigate the association between ferroptosis and pyroptosis and the upstream regulatory mechanisms. METHODS: This study included 30 patients with ALF and 30 healthy individuals who underwent serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) testing. C57BL/6 mice were also intraperitoneally pretreated with SIRT1, p53, or glutathione peroxidase 4 (GPX4) inducers and inhibitors and injected with lipopolysaccharide (LPS)/D-galactosamine (D-GalN) to induce ALF. Gasdermin D (GSDMD)-/- mice were used as an experimental group. Histological changes in liver tissue were monitored by hematoxylin and eosin staining. ALT, AST, glutathione, reactive oxygen species, and iron levels were measured using commercial kits. Ferroptosis- and pyroptosis-related protein and mRNA expression was detected by western blot and quantitative real-time polymerase chain reaction. SIRT1, p53, and GSDMD were assessed by immunofluorescence analysis. RESULTS: Serum AST and ALT levels were elevated in patients with ALF. SIRT1, solute carrier family 7a member 11 (SLC7A11), and GPX4 protein expression was decreased and acetylated p5, p53, GSDMD, and acyl-CoA synthetase long-chain family member 4 (ACSL4) protein levels were elevated in human ALF liver tissue. In the p53 and ferroptosis inhibitor-treated and GSDMD-/- groups, serum interleukin (IL)-1ß, tumour necrosis factor alpha, IL-6, IL-2 and C-C motif ligand 2 levels were decreased and hepatic impairment was mitigated. In mice with GSDMD knockout, p53 was reduced, GPX4 was increased, and ferroptotic events (depletion of SLC7A11, elevation of ACSL4, and iron accumulation) were detected. In vitro, knockdown of p53 and overexpression of GPX4 reduced AST and ALT levels, the cytostatic rate, and GSDMD expression, restoring SLC7A11 depletion. Moreover, SIRT1 agonist and overexpression of SIRT1 alleviated acute liver injury and decreased iron deposition compared with results in the model group, accompanied by reduced p53, GSDMD, and ACSL4, and increased SLC7A11 and GPX4. Inactivation of SIRT1 exacerbated ferroptotic and pyroptotic cell death and aggravated liver injury in LPS/D-GalN-induced in vitro and in vivo models. CONCLUSION: SIRT1 activation attenuates LPS/D-GalN-induced ferroptosis and pyroptosis by inhibiting the p53/GPX4/GSDMD signaling pathway in ALF.

[Effects of blueberry on the expression of PPARγ and PDGF-B in rat hepatic fibrosis].

OBJECTIVE: To explore the effects of blueberry on the expressions of peroxisome proliferator-activated receptor γ (PPARγ) and platelet-derived growth factor B (PDGF-B) in rat hepatic fibrosis. METHODS: A total of 45 male Sprague-Dawley rats were randomly divided into control group, CCl4-induced hepatic fibrosis (model group), blueberry prevention (BB group), DSHX prevention (DSHX group) and blueberry+DSHX prevention (BB+DSHX group) (n = 9 each). Fibrous liver models of rats were induced by subcutaneous injection of CCl4 and high-lipid/low-protein diet for 8 weeks except for control group. Then the expressions of collagen I (ColI), PPARγ and PDGF-B were determined by real-time polymerase chain reaction (RT-PCR), immunohistochemistry and Western blot respectively. RESULTS: Compared with the control group, the expression of PPARγ decreased and those of ColI and PDGF-B were elevated in model group (P < 0.05). The expression of PPARγ increased and ColI and PDGF-B decreased in BB, DSHX and BB+DSHX groups as compared to model group (P < 0.05). CONCLUSION: The inhibitory effects of blueberry in CCl4-induced liver fibrosis may be correlated with the activation of PPARγ, the inhibited expression of PDGF-B and the reduced synthesis of extracellular matrix.

[Effects of blueberry on hepatic fibrosis and ultrastructural of hepatocytes in rats].

OBJECTIVE: To observe the effects of blueberry on rat with hepatic fibrosis and ultrastructural. of hepatocytes. METHODS: Sixty (60) healthy Wistar rats were randomly divided into six groups: normal control group (group A), hepatic fibrosis model group (group B), blueberry at low, middle and high concentration groups (group C, D, E), Fu-Fang-Bie-Jia-Ruan-Gan tablet group (group F). The hepatic fibrosis model of rat was established by intraperitoneal injection of porcine serum once daily for 12 weeks. Simultaneously, rats in groups C-F were respectively perfused with blueberry juice or Fu-Fang-Bie-Jia-Ruan-Gan tablet for 12 weeks except for the normal control group which accepted saline alone. Upon terminal sacrifices of all rats at the end of the twelve weeks. Pathology of hepatic tissue was evaluated by hematoxylin-eosin (HE), Masson staining and transmission electron microscope. Liver index were measured. Levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were examined. Activities of superoxide dismutase (SOD), the contents of malondialdehyde (MDA), hydroxyproline (Hyp) and reduced glutathione (GSH) in liver homogenates were determined. RESULTS: Refer to the serum levels of ALT and AST, there is no significant difference existed in all groups (P > 0.05). Activities of SOD, the contents of GSH in liver homogenates of group D and E were significantly higher than those of group B(P < 0.05), while liver index, the contents of Hyp, MDA in liver homogenates were significantly lower than those of group B (P < 0.05). Compared with the group B, the pathological stages of hepatic fibrosis in group D and E were significantly reduced (P < 0.05), the expression of collagen were decreased, ultrastructural alterations were markedly improved. CONCLUSION: Blueberry may have certain preventional protective effects on porcine serum induced rat hepatic fibrosis and exhibit certain protective effects on organelles of hepatocytes (such as mitochondria), especially in middle and high does of blueberry. Its mechanism is probably related to increase the ability of antioxidant stress by increasing SOD activity and GSH content and reducing MDA content.

[Study of potential protective effects of blueberry on hepatic cytochrome P450 2E1 expression in rats with immune hepatic fibrosis].

OBJECTIVE: To study the protective effects of blueberry against rat immune hepatic fibrosis, specifically through the expression of hepatic cytochrome P450 2E1. METHOD: Fifty Wistar rats were randomly divided into five study groups (n = 10 each): Group A: normal control group, Group B: hepatic fibrosis model group, Group C: preventive group administered blueberry juice, Group D: preventive group administered Fu-Fang-Bie-Jia-Ruan-Gan tablet, and Group E: preventive group administered a combination of blueberry juice and Fu-Fang-Bie-Jia-Ruan-Gan tablet. The hepatic fibrosis model was established by intraperitoneal injection of porcine serum once daily for 12 weeks. Simultaneously, rats in preventive groups (Groups C-E) were perfused with blueberry juice or Fu-Fang-Bie-Jia-Ruan-Gan tablet or combinations of blueberry juice and Fu-Fang-Bie-Jia-Ruan-Gan tablet, respectively, for 12 weeks. The normal control group was perfused with saline for 12 weeks. All animals were sacrificed at the end of the 12 weeks, and serum levels of alanine aminotransferase (ALT) were measured and activities of superoxide dismutase (SOD), malondialdehyde (MDA), and hydroxyproline (Hyp) in liver homogenates were determined. Pathology of hepatic fibrosis was evaluated by hematoxylin-eosin (HE) and Masson staining. Expression of CYP2E1 was detected by real-time RT-PCR, immunohistochemical techniques, and Western blotting. RESULTS: Serum ALT levels were not significantly different in the control and treatment groups (F=4.056, P more than 0.05): A: 37.87+/-4.53 U/L, B: 49.23+/-9.81 U/L, C: 39.94+/-6.32 U/L, D: 40.50+/-5.70 U/L, and E: 38.24+/-8.43 U/L. Compared with Group B, the pathological stages of hepatic fibrosis were significantly reduced in the prevention groups (C-E) (F=95.097, P less than 0.05). Hyp and MDA in liver homogenates of groups C-E were significantly lower than those of Group B (Hyp: C: 472.68+/-44.14 mug/g, D: 416.12+/-39.38 mug/g, E: 429.51+/-55.14 mug/g vs. B: 603.16+/-68.92 mug/g, F=39.315, P less than 0.05; MDA: C: 0.83+/-0.06 nmol/mg, D: 0.96+/-0.08 nmol/mg, E: 0.85+/-0.06 nmol/mg vs. B: 1.24+/-0.15 nmol/mg, F=46.376, P less than 0.05). In contrast, SOD activities in Group C-E were significantly higher than those in Group B (C: 2.47+/-0.38 U/mg, D: 1.95+/-0.45 U/mg, E: 2.16+/-0.23 U/mg vs. B: 1.56+/-0.41 U/mg, F=25.557, P less than 0.05). Compared with Group A, the mRNA and protein expressions of CYP2E1 were increased in groups B-E, however the differences did not reach statistical significance (mRNA: F=0.897, protein: F=0.492, both P more than 0.05). The mRNA and protein expressions of CYP2E1 in groups C-E were lower than those of Group B, however the differences did not reach statistical significance (mRNA: F=0.897, protein: F=0.492, P more than 0.05). CONCLUSION: Blueberry exhibits certain protective effects against porcine serum-induced hepatic fibrosis in rats. The expression of hepatic cytochrome P450 2E1 in rats with immune hepatic fibrosis is not significantly different from the normal rats. Blueberry has no effect on the expression of hepatic cytochrome P4502E1.

[Effect of interferon and ribavirin combination therapy in sixty-two patients with chronic hepatitis C originating from a single blood donor].

To investigate the efficacy of interferon alpha 2 b plus ribavirin combination therapy in sixty-two patients with chronic hepatitis c (CHC) infection originating from a single blood donor. The 62 patients who developed CHC following blood transfusion from a known single infected donor were treated with interferon and ribavirin combination therapy for 48 weeks and followed-up for 96 weeks. The therapy regimen consisted of subcutaneous administration of 3-500 MIU interferon alpha 2 b every other day and daily oral administration of 0.6-1.0 g of ribavirin. Patients were monitored during treatment and in follow-up for sustained virological response (SVR), early virology response (EVR), treatment end virology response (ETVR), biochemical response of withdrawals, and side effects. The SVR rate was 83.9% (52/62). The EVR rate was 95.2% (59/62). The ETVR rate was 87.1% (54/62). The biochemical response rate after withdrawal of treatment was 100.0%. Eight patients developed mildly abnormal thyroid function as a result of the interferon therapy, but all were able to complete the antiviral treatment regimen under the care of endocrinologists. Younger age, relatively short course of disease, low viral load, and better compliance, but not sex, were correlated to curative effect of the combination therapy. Interferon alpha 2 b plus ribavirin combination therapy had a significant curative effect on a group of 62 CHC patients originating from a single case, with 52 of the patients showing SVR out to 96 weeks after therapy. Antiviral treatment is recommended for hepatitis C virus-positive patients to eradicate the virus and prevent disease progression.

[Eighty three cases of post transfusion HCV infection: a 10-year follow up].

OBJECTIVE: To find out clinical characteristics and natural history of post transfusion HCV infection. METHODS: 83 subjects who have received the blood from a same blood donor from January 1998 to July 2002 were investigated by the method combining cross-sectional study with retrospective study. HCV-antibody, HCV RNA, liver function, abdomen B-ultrasound and Fibroscan were detected. RESULTS: The HCV-antibody were all positive. The HCV RNA of 56 out of the 83 cases were positive. The chronicity rates of hepatis C were 76.3% (29/38) in male patients and 60.00% (27/45) in female patients respectively, without significant difference (X² = 2.99, P = 0.11). The average age of the HCV RNA positive patients was (36.54 ± 14.37) years old. The average age of the HCV RNA negative patients was (27.43 ± 12.51) years old. A significant difference (T = -2.41, P = 0.018) existed between. The HCV genotype was type1b. Among the HCV RNA positive patients,10 cases were with mild asthenia, anorexia and abdominal distention, 9 cases with increased serum ALT, 12 cases.with chronic hepatitis and 1 case was diagnosed with decompensated liver cirrhosis. CONCLUSION: The clinical manifestations of HCV infection are occult and chronic. The chronicity rate is related to gender and the age when infection was caught.

Transforming growth factor beta-1 upregulates glucose transporter 1 and glycolysis through canonical and noncanonical pathways in hepatic stellate cells.

BACKGROUND: Hepatic stellate cells (HSCs) are the key effector cells mediating the occurrence and development of liver fibrosis, while aerobic glycolysis is an important metabolic characteristic of HSC activation. Transforming growth factor-ß1 (TGF-ß1) induces aerobic glycolysis and is a driving factor for metabolic reprogramming. The occurrence of glycolysis depends on a high glucose uptake level. Glucose transporter 1 (GLUT1) is the most widely distributed glucose transporter in the body and mainly participates in the regulation of carbohydrate metabolism, thus affecting cell proliferation and growth. However, little is known about the relationship between TGF-ß1 and GLUT1 in the process of liver fibrosis and the molecular mechanism underlying the promotion of aerobic glycolysis in HSCs. AIM: To investigate the mechanisms of action of GLUT1, TGF-ß1 and aerobic glycolysis in the process of HSC activation during liver fibrosis. METHODS: Immunohistochemical staining and immunofluorescence assays were used to examine GLUT1 expression in fibrotic liver tissue. A Seahorse extracellular flux (XF) analyzer was used to examine changes in aerobic glycolytic flux, lactate production levels and glucose consumption levels in HSCs upon TGF-ß1 stimulation. The mechanism by which TGF-ß1 induces GLUT1 protein expression in HSCs was further explored by inhibiting/promoting the TGF-ß1/mothers-against-decapentaplegic-homolog 2/3 (Smad2/3) signaling pathway and inhibiting the p38 and phosphoinositide 3-kinase (PI3K)/AKT signaling pathways. In addition, GLUT1 expression was silenced to observe changes in the growth and proliferation of HSCs. Finally, a GLUT1 inhibitor was used to verify the in vivo effects of GLUT1 on a mouse model of liver fibrosis. RESULTS: GLUT1 protein expression was increased in both mouse and human fibrotic liver tissues. In addition, immunofluorescence staining revealed colocalization of GLUT1 and alpha-smooth muscle actin proteins, indicating that GLUT1 expression was related to the development of liver fibrosis. TGF-ß1 caused an increase in aerobic glycolysis in HSCs and induced GLUT1 expression in HSCs by activating the Smad, p38 MAPK and P13K/AKT signaling pathways. The p38 MAPK and Smad pathways synergistically affected the induction of GLUT1 expression. GLUT1 inhibition eliminated the effect of TGF-ß1 on HSC proliferation and migration. A GLUT1 inhibitor was administered in a mouse model of liver fibrosis, and GLUT1 inhibition reduced the degree of liver inflammation and liver fibrosis. CONCLUSION: TGF-ß1 induces GLUT1 expression in HSCs, a process related to liver fibrosis progression. In vitro experiments revealed that TGF-ß1-induced GLUT1 expression might be one of the mechanisms mediating the metabolic reprogramming of HSCs. In addition, in vivo experiments also indicated that the GLUT1 protein promotes the occurrence and development of liver fibrosis.

Effect of blueberry on hepatic and immunological functions in mice.

BACKGROUND: Conventional drugs used in the treatment and prevention of liver diseases often have side effects, therefore research into natural substances are of significance. This study examined the effects of blueberry on liver protection and cellular immune functions. METHODS: To determine the effects of blueberry on liver protective function, male mice were orally administered blueberry (0.6 g/10 g) or normal saline for 21 days. Hepatic RNA was extracted by Trizol reagent, and the expression of Nrf2, HO-1, and Nqo1 was determined by real-time RT-PCR. Superoxide dismutase (SOD) and malondialdehyde (MDA) in liver homogenate were determined, and liver index was measured. To assess the effects of blueberry on cellular immune function, male mice received blueberry (0.4, 0.6, or 0.8 g/10 g) for 35 days, and the percentages of CD3+, CD4+, and CD8+ T lymphocyte subgroups in peripheral blood were detected by flow cytometry, the index of the thymus and spleen was measured, and lymphocyte proliferation in the spleen was determined by MTT assay. RESULTS: Blueberry treatment significantly increased the expression of Nrf2, HO-1, and Nqo1, the important antioxidant components in the liver. Hepatic SOD in the blueberry group was higher and MDA was lower than that in the control group (P<0.05). Blueberry also increased the index of the spleen and enhanced the proliferation of lymphocytes of the spleen (P<0.05). The percentages of the CD3+ and CD4+ T lymphocyte subsets and the CD4+/CD8+ ratio were also increased by blueberry (P<0.05). CONCLUSIONS: Blueberry induces expression of Nrf2, HO-1, and Nqo1, which can protect hepatocytes from oxidative stress. In addition, blueberry can modulate T-cell function in mice.

[Effects of blueberry on the proliferation and activation of hepatic stellate cell and its mechanism].

OBJECTIVE: To investigate the effects of blueberry on the proliferation and activation of hepatic stellate cell (HSC) and its mechanism. METHODS: Rat HSC were isolated by type IV collagenase digestion and Nycodenz density gradient centrifugation. The cultured HSC was incubated at different concentrations of 10% blueberry serum. The 10% DSHX serum was used as positive control and 10% normal rat serum group as control. MTT colorimetric assay was used to detect the HSC proliferation. ColI of culture supernatant was detected by ELISA. The expression of α-SMA in HSC was measured by immunocytochemistry staining while the expressions of Nrf2 and HO-1 were determined by Western blot. RESULTS: Compared with controls, the low and high-dose blueberry serum groups significantly inhibited the HSC proliferation (P < 0.05). It had the same inhibitory effects as the positive control serum group (P > 0.05). ColI of culture supernatant obviously decreased (P < 0.05). And the expression levels of α-SMA in low and high-dose blueberry serum groups decreased significantly (P < 0.05). And there were similar effects in low & high-dose blueberry serum and positive control serum groups. Western blot showed that the expressions of Nrf2 and HO-1 in blueberry and positive control serum groups were higher than that in control group. And the increment was more significant in the low and high-dose blueberry serum groups (P < 0.05). CONCLUSION: Blueberry can significantly inhibit the proliferation and activation of HSC and reduce the synthesis of extra-cellular matrix. It may have potential preventive and protective effects on hepatic fibrosis. The mechanism may be related to an elevated expression of HO-1 through the Nrf2 pathway.

[Effects of blueberry on the expression patterns of heme oxygenase-1 in rats with hepatic fibrosis].

OBJECTIVE: To study the protective effect of Blueberry against rat hepatic fibrosis and the effect of Blueberry on HO-1 expression patterns. METHODS: A total of 45 SD rats were randomly divided into five groups namely control group (group A), model group (group B), blueberry group (group C), Dan-shao-hua-xian (DSHX) capsule group (group D) and blueberry +Dan-shao-hua-xian group (group E). Fibrous liver models in rats were induced by subcutaneous injection of CCl4 and high-lipid/low-protein diet for 8 weeks except the control group which accepted saline alone. The level of alanine aminotransferase (ALT) in serum was examined. The activities of superoxide dismutase (SOD) and malondialdehyde (MDA) in liver homogenates were determined. by the xanthine oxidase method and the thiobarbituric acid method. The pathology of hepatic fibrosis was evaluated by hematoxylin and eosin (H and E) staining. The Expression of HO-1 was detected by real-time RT-PCR, immunohistochemical techniques and western blotting. RESULTS: Serum ALT levels in every prevention group was lower than the group B [(149.44+/-16.51), (136.88+/-10.07), (127.38+/-11.03) vs (203.25+/-31.62) U/L, F = 92.498, P < 0.05], the SOD of liver homogenate in prevention group was significantly higher and the MDA was lower compared with the group B [SOD: (1.36+/-0.09), (1.42+/-0.13), (1.50+/-0.15) vs (1.08+/-0.19) U/mg, F = 13.671, P < 0.05; MDA: (0.294+/-0.026), (0.285+/-0.025), (0.284+/-0.028) vs (0.335+/-0.056) nmol/mg, F = 20.809, P < 0.05]. The pathological stages of hepatic fibrosis were all significantly reduced in prevention group (Chi2 test = 24.956, P < 0.05). Compared with group A, the mRNA and protein expressions of HO-1 were elevated (F = 4.549, 22.926, P < 0.05) in group B and increased in group C-E, but there is no significant difference existed. CONCLUSION: Blueberry may have preventive and protective effects on CCl4-induced hepatic fibrosis by reducing lipid peroxidation. However, these effects may not be related to the activation of HO-1 during long-term of CCl4.

Reduced delay in diagnosis of odontogenic keratocysts with malignant transformation: A case report.

BACKGROUND: In rare cases, odontogenic keratocysts (ODs) transform into squamous cell carcinoma. Intervals between the first attendance of a patient and the diagnosis of OD with malignant transformation vary from weeks to years. In this article, we report a case of malignancy derived from OD with a five-day delay in diagnosis. CASE SUMMARY: A 54-year-old woman was referred to Tongji Hospital in Wuhan, China with complaints of moderate pain, recurrent swelling, and pus discharge around her left maxillary lateral incisor for over 10 years. Physical examination revealed a fistula at the palatine-side mucoperiosteum of the left maxillary lateral incisor and enlarged lymph node in the left neck. Cone beam computed tomography revealed a cystic lesion with massive bone destruction from the left maxillary central incisor to the left secondary maxillary premolar and local bony destruction in the left first mandibular molar. The patient was clinically diagnosed with OD. Enucleation rather than marsupialization was performed given the risk factors of long history, recent aggravated pain, and massive bony destruction. Malignant transformation of OD was confirmed by pathologists 3 d after the operation. Radical surgery was performed, and lymph node metastasis was observed. The patient was subjected to postoperative radiotherapy and synchronous chemotherapy, and no local recurrence or distant metastasis was noted at one-year follow-up. CONCLUSION: Our case suggests that clinicians should be aware of the malignant transformation of OD, especially when patients present with a long history, massive cyst, chronic inflammation, recent persistent infections, aggravated pain, numbness around the cystic lesion, and lymph node enlargement.

Platycodin D alleviates liver fibrosis and activation of hepatic stellate cells by regulating JNK/c-JUN signal pathway.

Liver fibrosis is involved in the progression of most chronic liver diseases. Even though we have made a huge progress in order to understand the pathogenesis of liver fibrosis, however, there is still a lack of productive treatments. Being a traditional Chinese medicine, Platycodin D (PD), an oleanane kind of triterpenoid saponin has been put to extensive use for treating different kinds of illnesses that include not just anti-nociceptive, but also antiviral, anti-inflammatory, and anti-cancer for thousands of years. Nonetheless, there has been no clarification made for its effects on the progression of liver fibrosis. In this manner, we carried out in vitro studies for the purpose of investigating the anti-fibrosis impact of PD. Activation of hepatic stellate cells was evaluated by means of the detection of the proliferation of HSCs and the expression of specific proteins. We discovered the fact that PD had the potential of activating HSCs. Thereafter, we detected the apoptosis and autophagy of the HSCs; as the results suggested, PD induced apoptosis and autophagy of the HSCs. It augmented the expression level of apoptotic proteins that included Bax, Cytochrome C (cyto-c), cleaved caspase3 and cleaved caspase9, in addition to the autophagy relevant proteins, for instance, LC3II, beclin1, Atg5 and Atg9. Further research was carried out for the investigation of the underlying molecular mechanism, and discovered that PD promoted the phosphorylation of JNK and c-Jun. Treating the JNK inhibitor P600125 inhibited the effect of PD, confirming the impact of PD on the regulation of JNK/c-Jun pathway. Thus, we speculated that PD alleviates liver fibrosis and activation of hepatic stellate via promoting phosphorylation of JNK and c-Jun and further altering the autophagy along with apoptosis of HSCs.

[Study on blood and buccal mucous related genes expression in the patients suffering from coal-burning arsenism].

OBJECTIVE: To observe the oxidative stress and skin disease related genes expression in cells exfoliated from buccal mucous and blood in patients with arseniasis caused by coal-burning pollution in Guizhou. METHODS: Buccal and blood cell samples were collected from arsenicosis patients and healthy subjects residing in Guizhou. Total RNA was isolated for related genetic transcript analysis. Real-time PCR (RT-PCR) was employed to detect the change of related gene expression. Serum indicators of liver injury, such as serum alanine aminotransferase (ALT), were detected. RESULTS: Serum levels of ALT in arsenic patients were significantly higher than those in controls (36.9 +/- 17.4 / 25.8 +/- 2.4, t = 0.019, P < 0.05) even when no symptom was found in these patients. Buccal K10 gene was overexpressed (3.6 +/- 0.9 / 1.8 +/- 0.7, t = 0.041, P < 0.05) and MT-1 expression was significantly lower (0.8 +/- 0.5 / 3.9 +/- 0.4, t = 0.028, P < 0.05) in arsenic patients than that in control while the expressions of other genes were the same as those of controls. CONCLUSION: The expressions of MT-1 and K10 gene in buccal mucous cells may be regarded as sensitive molecular markers for skin and hepatic pathologic changes in arsenic patients. Protection of liver function should be considered as an important step in arseniasis prevention and control.

[Identification of differential proteins of serum in the patients suffering from coal-burning arsenism].

OBJECTIVE: To compare and analyze the differential expression of proteins between coal-burning arsenism serum and normal human serum and identify the proteins related with arseniasis caused by coal-burning. METHODS: Serum samples were collected from 6 normal subjects and 6 patients suffering from coal-burning arsenism. 2-DE was performed to separate serum proteins. After silver staining, the differential expression of proteins was analyzed and then identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). RESULTS: There were an average of 779 +/- 35 spots and 865 +/- 30 spots on 2-DE matching of two groups and the matching rate was 90.1% between two groups. From these two groups, 60 different protein spots were identified. Up-regulated expression was observed in 25 proteins and down-regulated expression in 35 proteins in the patient serum group. Among which 35 with differential expression above three times were singled out and MALDI-TOF-MS analysis was carried out on them. Thirteen proteins were identified, including keratin 10, apolipoprotein A-V, transferrin, alpha-1-antitrypsin, human zinc-alpha-2-glycoprotein, mitogen-activated protein kinase 3, vacuolar protein sorting 33A, O-linked GlcNAc transferase and etc. Up-regulated expression was observed in 5 proteins and down-regulated expression in 8 proteins in the patient serum group. CONCLUSION: The well-resolved and reproducible 2-DE serum patterns of patients suffering from coal-burning arsenism were established and some differentially expressed proteins characterized. These data will be used to screen the biomarker and to further study arseniasis caused by coal-burning.

[Model establishment of liver fibrosis in oral arsenic solution exposed mice].

OBJECTIVE: To discuss and compare the model establishment of liver fibrosis in oral arsenic solution exposed mice and mice with high-fat feedstuff. METHODS: A total of 240 mice were divided randomly into 6 groups: control group, sodium arsenite group, sodium arsenate group, high-fat feedstuff group, sodium arsenite group with high-fat feedstuff and sodium arsenate group with high-fat feedstuff with 40 mice each. Control group and high-fat feedstuff group (drinking tap water), sodium arsenite group and sodium arsenite group with high-fat feedstuff (drinking 300 mg/L iAs3+ water), sodium arsenate group and sodium arsenate group with high-fat feedstuff (drinking 300 mg/L iAs5+ water). The mice were sacrificed after 3, 6, 10 months' arsenic-exposure and examined for liver function. HE dyeing and Masson dyeing were also employed to observe the pathological changes in hepatic tissue in each group. RESULTS: After 3 months' modeling, ALT and AST in control group, sodium arsenite group, sodium arsenate group, sodium arsenite group with high-fat feedstuff and sodium arsenate group with high-fat feedstuff were (36.7 +/- 5.7) U/L and (110 +/- 22) U/L, (55.6 +/- 4.6) U/L and (249 +/- 41) U/L, (52.6 +/- 8.8) U/L and (161 +/- 15) U/L, (311.3 +/- 19.7) U/L and (484 +/- 15) U/L and (515.0 +/- 60.8) U/L and (671 +/- 24) U/L. They were higher in all the arsenic groups than in control group (P < 0.05); all the HE dyeing samples in arsenic groups showed liver injury in varying degrees such as hydropic degeneration, fatty degeneration, spotty necrosis, focal necrosis and inflammatory cell infiltration. There were liver cell regeneration and fibroplasia in varying degrees. The liver injury of the mice in all arsenic groups aggravated as exposure time prolonged. Masson dyeing after 10 months' modeling showed hyperplasia in portal areas and central venous areas; the mean area of fibrosis in control group, sodium arsenite group, sodium arsenate group, sodium arsenite group with high-fat feedstuff and sodium arsenate group with high-fat feedstuff were 0.1333, 0.5584, 0.5250, 0.7534 and 0.7200 respectively. There was statistical significance between arsenic groups and control group (P < 0.05) CONCLUSION: The liver injury and fibrosis model in oral arsenic solution exposed mice and those with high-fat feedstuff are successfully established and subsequently evaluated. It is a comparatively ideal animal model for studying arsenic liver injury and fibrosis.