Pacenta polypeptide injection alleviates the fibrosis and inflammation in cigarette smoke extracts-induced BEAS-2B cells by modulating MMP-9/TIMP-1 signaling.

Chronic obstructive pulmonary disease (COPD) has high morbidity and mortality. Here, we aimed to explore the roles and potential correlation of placenta polypeptide injection (PPI) and MMP-9/TIMP-1 signaling pathway in COPD. BEAS-2B cells were treated with cigarette smoke extract (CSE) to establish a COPD cell model in vitro. The cell survival and cytotoxic effect were measured by CCK-8, LDH release and flow cytometry assays. The inflammatory responses were determined by western blot and ELISA assay. Cell fibrosis was assessed by immunofluorescence and western blot assays. PPI treatment had no cytotoxic effect on BEAS-2B cells until the final concentration reached to 10%. In the range of 0%-8% final concentration, PPI treatment weakened CSE-induced the decrease of cell viability and the increase of LDH level in a concentration-dependent manner. Four percent PPI treatment enhanced cell viability and decreased cell apoptosis of CSE-treated cells in a time-dependent manner. Moreover, 4% PPI treatment significantly decreased inflammatory responses and fibrosis induced by CSE, while AMPA (MMPs agonist) had opposite effects. Notably, AMPA reversed the protective roles of PPI on CSE-induced inflammation and fibrosis. Mechanistically, 4% PPI treatment significantly suppressed MMP-1, MMP-2, MMP-3, MMP-9, MMP-13, and MMP-19 levels, but enhanced TIMP-1, TIMP-2, TIMP-3, and TIMP-4 levels. Among them, MMP-9 and TIMP-1 might be the main target of PPI. PPI effectively attenuated CSE-induced inflammation and fibrosis in vitro by regulating MMP-9/TIMP-1 signaling pathway.

Hydrogel-Based, Dynamically Tunable Plasmonic Metasurfaces with Nanoscale Resolution.

Flat metasurfaces with subwavelength meta-atoms can be designed to manipulate the electromagnetic parameters of incident light and enable unusual light-matter interactions. Although hydrogel-based metasurfaces have the potential to control optical properties dynamically in response to environmental conditions, the pattern resolution of these surfaces has been limited to microscale features or larger, limiting capabilities at the nanoscale, and precluding effective use in metamaterials. This paper reports a general approach to developing tunable plasmonic metasurfaces with hydrogel meta-atoms at the subwavelength scale. Periodic arrays of hydrogel nanodots with continuously tunable diameters are fabricated on silver substrates, resulting in humidity-responsive surface plasmon polaritons (SPPs) at the nanostructure-metal interfaces. The peaks of the SPPs are controlled reversibly by absorbing or releasing water within the hydrogel matrix, the matrix-generated plasmonic color rendering in the visible spectrum. This work demonstrates that metasurfaces designed with these spatially patterned nanodots of varying sizes benefit applications in anti-counterfeiting and generate multicolored displays with single-nanodot resolution. Furthermore, this work shows system versatility exhibited by broadband beam-steering on a phase modulator consisting of hydrogel supercell units in which the size variations of constituent hydrogel nanostructures engineer the wavefront of reflected light from the metasurface.

The monomer TEC of blueberry improves NASH by augmenting tRF-47-mediated autophagy/pyroptosis signaling pathway.

BACKGROUND: Nonalcoholic steatohepatitis (NASH) is one of the most common liver diseases and has no safe and effective drug for treatment. We have previously reported the function of blueberry, but the effective monomer and related molecular mechanism remain unclear. METHODS: The monomer of blueberry was examined by ultra performance liquid chromatography-mass spectrometry (UPLC-MS). The NASH cell model was constructed by exposing HepG2 cells to free fatty acids. The NASH mouse model was induced by a high-fat diet for 12 weeks. NASH cell and mouse models were treated with different concentrations of blueberry monomers. The molecular mechanism was studied by Oil Red O staining, ELISA, enzyme activity, haematoxylin-eosin (H&E) staining, immunohistochemistry, immunofluorescence, western blot, RNA sequencing, and qRT-PCR. RESULTS: We identified one of the main monomer of blueberry as tectorigenin (TEC). Cyanidin-3-O glucoside (C3G) and TEC could significantly inhibit the formation of lipid droplets in steatosis hepatocytes, and the effect of TEC on the formation of lipid droplets was significantly higher than that of C3G. TEC can promote cell proliferation and inhibit the release of inflammatory mediators in NASH cell model. Additionally, TEC administration provided a protective role against high-fat diets induced lipid damage, and suppressed lipid accumulation. In NASH mouse model, TEC can activate autophagy, inhibit pyroptosis and the release of inflammatory mediators. In NASH cell model, TEC inhibited pyroptosis by stimulating autophagy. Then, small RNA sequencing revealed that TEC up-regulated the expression of tRF-47-58ZZJQJYSWRYVMMV5BO (tRF-47). The knockdown of tRF-47 blunted the beneficial effects of TEC on NASH in vitro, including inhibition of autophagy, activation of pyroptosis and release of inflammatory factors. Similarly, suppression of tRF-47 promoted the lipid injury and lipid deposition in vivo. CONCLUSIONS: These results demonstrated that tRF-47-mediated autophagy and pyroptosis plays a vital role in the function of TEC to treat NASH, suggesting that TEC may be a promising drug for the treatment of NASH.

Outcomes of Long-term Treatment of Chronic HBV Infection With Entecavir or Other Agents From a Randomized Trial in 24 Countries.

BACKGROUND & AIMS: Treatment of chronic hepatitis B virus (HBV) infection with entecavir suppresses virus replication and reduces disease progression, but could require life-long therapy. To investigate clinical outcome events and safety associated with long-term treatment with entecavir, we followed up patients treated with entecavir or another standard-of-care HBV nucleos(t)ide analogue for up to 10 years. We assessed long-term outcomes and relationships with virologic response. METHODS: Patients with chronic HBV infection at 299 centers in Asia, Europe, and North and South America were assigned randomly to groups that received entecavir (n = 6216) or an investigator-selected nonentecavir HBV nucleos(t)ide analogue (n = 6162). Study participants were followed up for up to 10 years in hospital-based or community clinics. Key end points were time to adjudicated clinical outcome events and serious adverse events. In a substudy, we examined relationships between these events and virologic response. RESULTS: There were no significant differences between groups in time to event assessments for primary end points including malignant neoplasms, liver-related HBV disease progression, and death. There were no differences between groups in the secondary end points of nonhepatocellular carcinoma malignant neoplasms and hepatocellular carcinoma. In a substudy of 5305 patients in China, virologic response, regardless of treatment group, was associated with a reduced risk of liver-related HBV disease progression (hazard ratio, 0.09; 95% CI, 0.038-0.221) and hepatocellular carcinoma (hazard ratio, 0.03; 95% CI, 0.009-0.113). Twelve patients given entecavir (0.2%) and 50 patients given nonentecavir drugs (0.8%) reported treatment-related serious adverse events. CONCLUSIONS: In a randomized controlled trial of patients with chronic HBV infection, we associated entecavir therapy with a low rate of adverse events over 10 years of follow-up evaluation. Patients receiving entecavir vs another nucleos(t)ide analogue had comparable rates of liver- and non-liver-related clinical outcome events. Participants in a China cohort who maintained a virologic response, regardless of treatment group, had a reduced risk of HBV-related outcome events including hepatocellular carcinoma. ClinicalTrials.gov identifier no: NCT00388674.

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.

Thymosin-ß4 Mediates Hepatic Stellate Cell Activation by Interfering with CircRNA-0067835/miR-155/FoxO3 Signaling Pathway.

BACKGROUND/AIMS: Hepatic stellate cells (HSCs) are the primary cell type responsible for liver fibrosis. Our study proved that thymosin beta 4 (Tß4) has anti-fibrogenic effects in HSCs through PI3K/AKT pathway. However, the underlying mechanisms are not fully elucidated. Circular RNAs (circRNAs) play important roles in fine-tuning gene expression and are often deregulated in cancers. However, the expression profile and clinical significance of in liver fibrosis is still unknown. Therefore, we hypothesize that Tß4 influences circRNAs in liver fibrosis. METHODS: Circular RNA microarray was conducted to identify Tß4-related circRNAs. Pathway analysis and miRNA response elements analysis was conducted to predict the potential roles of differentially expressed circRNAs in liver fibrosis. CCK8 assays and flow cytometric assays were conducted to clarify the role of circRNA in liver fibrosis. Bioinformatics analysis and in vitro experiments were conducted to clarify the mechanism of circRNA-mediated gene regulation in liver fibrosis. RESULTS: A total of 644 differentially expressed circRNAs were identified between the Tß4-depleted LX-2 cells and the control LX2 cells. The expression of circRNA-0067835 was significantly increased in the Tß4-depleted LX-2 cells compared with control. Knockdown of circRNA-0067835 observably decreased LX-2 cell proliferation by causing G1 arrest and promoting apoptosis. Bioinformatics online programs predicted that circRNA-0067835 acted as miR-155 sponge to regulate FOXO3a expression, which was validated using luciferase reporter assay. CONCLUSION: Our experiments showed that circRNA-0067835 regulated liver fibrosis progression by acting as a sponge of miR-155 to promote FOXO3a expression, indicating that circRNA-0067835 may serve as a potential therapeutic target for patients with liver fibrosis.

AST to ALT ratio and arterial stiffness in non-fatty liver Japanese population:a secondary analysis based on a cross-sectional study.

BACKGROUND: Previous studies have revealed that triglyceride to high-density lipoprotein cholesterol (HDL-C) ratio (henceforth TG/HDL-C) is one of the major risk factors for cardiovascular disease, insulin resistance and metabolism syndrome. However, there are fewer investigations of the correlations between the aspartate aminotransferase (AST) to alanine aminotransferase (ALT) ratio and brachial ankle pulse wave velocity (baPWV). This study was undertaken to investigate the relationship between the AST to ALT ratio and brachial-ankle pulse wave velocity (baPWV) in a Japanese population. METHODS: The present study was a cross-sectional study. A total of 646 Japanese men and women without fatty liver, aged 24-84 years old, received a health medical check-up programme including the results from baPWV inspection and various standardized questionnaires in a health examination centre in Japan. Main outcome measures included AST/ALT ratio, baPWV, fatty liver and postmenopausal status. Abdominal ultrasonography was used to diagnose fatty liver. A postmenopausal state was defined as beginning 1 year following the cessation of menses. RESULTS: After adjusting for potential confounders (age, sex, BMI, SBP, DBP, AST, ALT, GGT, uric acid, fasting glucose, TC, LDL, eGFR, smoking and exercise statuses, fatty liver, alcohol consumption and ABI), a non-linear relationship was detected between AST/ALT and baPWV, which had an inflection point of 5.6. The effect sizes and the confidence intervals on the left and right sides of the inflection point were 12.7 (1.9 to 23.5) and - 16.7 (- 36.8 to 3.3), respectively. Subgroup analysis in participants with excessive alcohol consumption (more than 280 g/week) showed that AST/ALT had a negative correlation with baPWV (ß = - 30.7, 95%CI (- 53.1, - 8.4)), and the P value for the interaction was less than 0.05. CONCLUSION: The relationship between AST/ALT and baPWV is non-linear. AST/ALT was positively correlated with baPWV when AST/ALT was less than 5.6. In addition, the trend was the opposite in subjects who consumed excessive amounts of alcohol.

[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.

[Effects of blueberry on apoptosis and expression of Bcl-2 and Bax in HSC-T6].

OBJECTIVE: To investigate the effects of blueberry on the apoptosis, expression of Bcl-2 and Bax in rat hepatic stellate cell (HSC-T6). METHODS: 10% blueberry serum at low, middle and high dose, 10% Fu-Fang-Bie-Jia-Ruan-Gan tablet serum and 10% saline serum were prepared by method of serum pharmacology. Subcultured HSC-T6 was divided into saline serum control group, blueberry serum at low, middle, high dose and Fu-Fang-Bie-Jia-Ruan-Gan tablet serum group, and then was respectively incubated at different dose of 10% blueberry serum, 10% Fu-Fang-Bie-Jia-Ruan-Gan tablet serum and 10% saline serum for 72 hours.Apoptosis of HSC-T6 was detected using flow cytometry with annexin V FITC/PI double staining. The expression of Bcl-2 and Bax in HSC-T6 were examined using immunocytochemistry and Western blotting, respectively. RESULT: There was no significant difference for HSC-T6 Bax protein expression in the low, middle and high dose blueberry serum groups, compared with saline serum control group, respectively.In the high-dose blueberry serum group HSC-T6 early and total apoptosis rate increased significantly compared with the saline serum control group (5.55% ± 0.98% vs 2.53% ± 0.46%, 7.01% ± 1.05% vs 2.96% ± 0.81%, both P<0.05); Bcl-2 protein expression was significantly decreased (A value, 82 ± 35 vs 51 ± 13, P<0.05); Bcl-2/Bax ratio was significantly decreased (0.26 ± 0.02 vs 0.46 ± 0.03, P<0.05); HSC-T6 early and total apoptosis rate, Bcl-2 expression and Bcl-2/Bax ratio in the low and the middle dose blueberry serum group showed no significant difference with the saline serum control group. CONCLUSION: Blueberry can induce HSC-T6 apoptosis by down-regulating Bcl-2 expression and decreasing the ratio of Bcl-2/Bax in HSC-T6 cells, so it may have potential interference effects on hepatic fibrosis.

[Natural clearance of hepatitis C virus in 96 patients with infection acquired by blood transfusion from a single donor in Guizhou].

OBJECTIVE: To investigate the clinical features and rate of natural viral clearance in patients with hepatitis C virus (HCV) infection acquired by blood transfusion from a single donor. METHODS: Ninety-six patients who acquired HCV infection between January 1998 and December 2002, upon receipt of donated blood from a single infected individual in Guizhou,were included in this retrospective cross-sectional study. Patients were clinically assessed to determine levels of anti-HCV antibodies, HCV RNA and biochemical indicators of liver function,as well as features of liver structure (by abdominal B ultrasonography and elastography). HCV genetic testing was used to determine the virus genotype. Measurement data were expressed as mean ± standard deviation. Count data were analyzed by the x² test,with P less than 0.05 indicating statistical significance. RESULTS: All 96 patients tested positive for antiHCV antibodies. The majority of patients (70%; 34:33 male:female) had HCV RNA more than or equal to 1.0 * 103 copies/ml. All patients carried the same HCV genotype as the single blood donor:genotype lb. The overall rate of natural HCV clearance was 30.2%. but males had a significantly lower rate (19.0% (8/42) vs. females:38.9% (21/54);x²=4.41,P=0.023) as did older patients (more than 40 years-old:16.1% (5/31) vs .less than or equal to 40 years-old:36.9% (24/65);x²=4.30,P=0.028). The overall rate of chronic HCV infection (CHC) was 69.8%,but the rate was significantly lower in younger patients (less than or equal to 40 years-old:63.1% (41/65) vs. more than 40 years-old:83.9% (26/31);x²=6.67,P=0.028). Among the 67 patients with CHC,12 had symptoms of mild weakness,anorexia and abdominal distention,11 had elevated serum alanine aminotransferase (116.25 +/- 24.65 U/L) and stage 3 or 4 fibrosis (liver elasticity values more than or equal to 5.1 kPa),and three had mildly abnormal serum bilirubin (32.56 ± 5.28 mumol/L). Fifteen patients showed signs of chronic hepatitis and one patient showed signs of cirrhosis by abdominal B ultrasonography. None of the patients showed signs of hepatocellular carcinoma. CONCLUSION: The course of blood transfusion acquired HCV infection is largely unknown and natural viral clearance rate may be associated with sex-and age-related factors.

[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.

[A case-control study on the risk factors of hepatocellular carcinoma in Guizhou Province].

OBJECTIVE: To determine the risk factor of HCC in Guizhou. METHODS: A group case-control study design was conducted between 762 cases and 798 controls in Guizhou province. The main related-factors were analyzed with unconditional logistic regression model and evaluated by odds ratio (OR) and 95% confidence interval (95% CI). RESULTS: There are significant differences between cases and controls in regarding to cigarette smoking 210 (27.6%),non-alcoholic fatty liver disease 336 (44.1%), alcoholic liver disease 245 (32.2%), family history of HCC 141 (16.5%), alcohol consumption 300 (39.4%), HBV infection 436 (57.2%), pickled food 290 (38.1%), and economic status 5 years ago 420 (55.1%) in cases,and cigarette smoking 116 (14.5%),non-alcoholic fatty liver disease 160 (20.1%), alcoholic liver disease 101 (12.7%), family history of HCC 40 (5.0%), alcohol consumption 180 (22.6%), HBV infection 82 (10.3%), pickled food 225 (28.2%), and economic status 5 years ago 647 (81.1%) in controls, with OR of each variable was 3.520, 2.464, 4.330, 2.219, 2.451, 19.245, 6.212, 0.174 respectively, P less than 0.01. CONCLUSION: HBV infection and pickled food were the most common risks for HCC in Guizhou. Alcohol consumption excessively and cigarette smoking may increase the risk too.

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.

Prognosis of male lung cancer patients with urinary cancer: a study from a national population-based analysis.

Lung cancer accounts for the most cancer-related deaths in the world. Our previous study suggested the improved survival of lung cancer patients, mainly female patients, with subsequent metachronous primary breast cancer. However, whether the survival advantages of the two primaries are associated with patients' sex and the specific breast cancer is unclear. Whether male lung cancer patients with another primary may encounter the same survival advantage as female patients is also uncertain. The uncertainty hinders these patients from the potential benefit of lung cancer clinical trial. A total of 343 male lung adenocarcinoma patients with subsequent bladder papillary transitional cell carcinoma (LCBC), 1539 lung adenocarcinoma patients with prior bladder papillary transitional cell carcinoma (BCLC), 1181 lung adenocarcinoma patients with subsequent prostate adenocarcinoma (LCPC), 7426 lung adenocarcinoma patients with prior prostate adenocarcinoma (PCLC), and patients with single bladder/prostate/lung (SLC) cancer were identified from the Surveillance, Epidemiology, and End Results. Patients were classified into simultaneous two primary cancer (sTPC), metachronous two primary cancer 1 (mTPC1), or mTPC2 groups when interval time between two cancers was within 6 months, between 7 and 60 months, or over 60 months, respectively. Propensity matching score program was executed to match the two primary cancers with single primary. Cox regression and competing risk regression were performed to identify confounders associated with all-cause and cancer-specific survival, respectively. The major cancer-related and non-cancer-related death in the two primaries were lung cancer and heart disease, respectively. Median overall survival times since lung primary of LCBC and SLC were 97 and 17 months, respectively, and incidence of all-cause and cancer-specific death in LCBC since lung malignancy was significantly lower (Coef. - 1.24, 95% CI - 1.49 to 0.99; SHR 0.42, 95% CI 0.33-0.53). Among the categorized groups, prognosis values of sTPC and mTPC2 groups were not statically different from that of the matched single lung cancer, whereas increased overall survival time and decreased incidence of all-cause and cancer-specific death relative to the matched patients were observed in mTPC1 group (H.R 0.28, 95% CI 0.19-0.41; SHR 0.33, 95% CI 0.23-0.47). Similar prognosis of LCPC relative to SLC was also observed. Furthermore, a generally improved survival relative to SLC was observed in PCLC (median survival times of PCLC and SLC were 17 and 12 months, respectively; Coef. - 0.32, 95% CI - 0.43 to 0.22; SHR 0.77, 95% CI 0.69-0.85), whereas prognosis of BCLC was similar to the matched ones. These results hinted that survival of lung cancer patients might vary with prior cancer history. Further analysis among groups with the two primaries suggested that advanced bladder cancer was not associated with prognosis of patients with LCBC and BCLC. On the contrary, advanced prostate cancer was associated with all-cause and cancer-specific death in patients with PCLC but not in patients with LCPC. Compared with patients with single lung cancer, male lung cancer patients with subsequent bladder/prostate primary over 6 months experienced generally improved survival. These results were similar to our previous study regarding female lung cancer patients with another breast primary. On the contrary, male lung cancer patients with prior primary malignancy encountered varied prognosis: improved survival relative to single lung primary was observed in lung cancer with prior prostate cancer, whereas prognosis of lung cancer with prior bladder cancer was not different. Therefore, great attention was required to characterize prognosis of lung cancer patients with another primary in advance, which was essential to eliminate the potential bias when these patients were included into the clinical trials.

Human placental extract suppresses mast cell activation and induces mast cell apoptosis.

BACKGROUND: Human placental extract (HPE) has been documented to facilitate the healing of certain disorders including allergy. However, the effects of HPE on the functionality of mast cells, a critical cell type in allergic diseases, have not been reported. METHODS: To investigate the effects of HPE on the regulation of allergy with respect to the biological functions of mast cells, the mast cell line C57 or HMC-1 cells were treated with HPE followed by the assessment of cell proliferation, apoptosis, activation, chemotaxis and phagocytosis. Mouse peritoneal mast cells were also investigated for their responses to induction of apoptosis by HPE in vivo. Furthermore, the effect of HPE on mast cell degranulation was confirmed using the passive cutaneous anaphylaxis (PCA) assay, an acute allergy model. RESULTS: HPE was capable of suppressing mast cell proliferation and inducing mast cell apoptosis. Mast cell degranulation in response to compound 48/80- or anti-DNP IgE and DNP-mediated activation was suppressed. In addition, treatment with HPE compromised the production of cytokines by mast cells and cell chemotaxis. These observations were consistent with the dampened passive cutaneous anaphylaxis (PCA) assay following treatment with HPE. CONCLUSION: This study revealed a suppressive effect of HPE on overall mast cell activities, suggesting a potential regulatory role of HPE on the alleviation of allergic diseases through mast cells.

[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.