An-Luo-Hua-Xian Pill Improves the Regression of Liver Fibrosis in Chronic Hepatitis B Patients Treated with Entecavir.

Background and Aims: Chronic hepatitis B (CHB) can cause liver fibrosis and lead to cirrhosis and cancer. As the effectiveness of antiviral therapy to reverse liver fibrosis is limited, We aimed to evaluate the effect of An-Luo-Hua-Xian pill (ALHX) on fibrosis regression in CHB patients treated with entecavir (ETV). Methods: Treatment-naïve patients with CHB were randomly treated with ETV alone or combined with ALHX (ETV+ALHX) between October 1, 2013 and December 31, 2020. Demographic, laboratory, and liver histology data before and after 78 weeks of treatment were collected. The Ishak fibrosis score (F) was used and fibrosis regression required a decrease in F of ≥1 after treatment. Results: A total of 780 patients were enrolled, and 394 with a second liver biopsy after treatment were included in the per-protocol population, 132 in ETV group and 262 in ETV+ALHX group. After 78 weeks of treatment, the fibrosis regression rate in the ETV+ALHX group was significantly higher than that of the ETV group at baseline F≥3 patients: 124/211 (58.8%) vs. 45/98 (45.9%), p=0.035. The percentage of patients with a decreased liver stiffness measurement (LSM) was higher in the ETV+ALHX group: 156/211 (73.9%) vs. 62/98 (63.%), p=0.056. Logistic regression analysis showed that ETV combined with ALHX was associated with fibrosis regression [odds ratio (OR)=1.94, p=0.018], and a family history of hepatocellular carcinoma was on the contrary. (OR=0.41, p=0.031). Conclusions: ETV combined with ALHX increased liver fibrosis regression in CHB patients.

Dysregulated hepatic lipid metabolism and gut microbiota associated with early-stage NAFLD in ASPP2-deficiency mice.

Background: Growing evidence indicates that lipid metabolism disorders and gut microbiota dysbiosis were related to the progression of non-alcoholic fatty liver disease (NAFLD). Apoptosis-stimulating p53 protein 2 (ASPP2) has been reported to protect against hepatocyte injury by regulating the lipid metabolism, but the mechanisms remain largely unknown. In this study, we investigate the effect of ASPP2 deficiency on NAFLD, lipid metabolism and gut microbiota using ASPP2 globally heterozygous knockout (ASPP2+/-) mice. Methods: ASPP2+/- Balb/c mice were fed with methionine and choline deficient diet for 3, 10 and 40 day to induce an early and later-stage of NAFLD, respectively. Fresh fecal samples were collected and followed by 16S rRNA sequencing. HPLC-MRM relative quantification analysis was used to identify changes in hepatic lipid profiles. The expression level of innate immunity-, lipid metabolism- and intestinal permeability-related genes were determined. A spearman's rank correlation analysis was performed to identify possible correlation between hepatic medium and long-chain fatty acid and gut microbiota in ASPP2-deficiency mice. Results: Compared with the WT control, ASPP2-deficiency mice developed moderate steatosis at day 10 and severe steatosis at day 40. The levels of hepatic long chain omega-3 fatty acid, eicosapentaenoic (EPA, 20:5 n-3) and docosahexaenoic (DHA, 22:6 n-3), were decreased at day 10 and increased at day 40 in ASPP+/- mice. Fecal microbiota analysis showed significantly increased alpha and beta diversity, as well as the composition of gut microbiota at the phylum, class, order, family, genus, species levels in ASPP2+/- mice. Moreover, ASPP-deficiency mice exhibited impaired intestinal barrier function, reduced expression of genes associated with chemical barrier (REG3B, REG3G, Lysozyme and IAP), and increased expression of innate immune components (TLR4 and TLR2). Furthermore, correlation analysis between gut microbiota and fatty acids revealed that EPA was significantly negatively correlated with Bifidobacterium family. Conclusion: Our findings suggested that ASPP2-deficiency promotes the progression of NAFLD, alterations in fatty acid metabolism and gut microbiota dysbiosis. The long chain fatty acid EPA was significantly negatively correlated with Bifidobacterial abundance, which is a specific feature of NAFLD in ASPP2-deficiency mice. Totally, the results provide evidence for a mechanism of ASPP2 on dysregulation of fatty acid metabolism and gut microbiota dysbiosis.

Long-Term Carbohydrate-Containing Late-Evening Snack Significantly Improves the Ratio of Branched Chain Amino Acids to Aromatic Amino Acids in Adults with Liver Cirrhosis due to Hepatitis B.

BACKGROUND: The liver is the primary organ for amino acid metabolism, and metabolic disorder of amino acids is common in liver disease. However, the characteristics of plasma amino acid profiles in patients with HBV-related cirrhosis and the impacts of late-evening snack (LES) on cirrhosis are unclear. OBJECTIVES: To investigate the characteristics of plasma amino acid profiles in patients with HBV-related chronic hepatitis, cirrhosis, and the effects of late-evening snacks on plasma amino acid profiles. METHODS: 86 patients with HBV-related cirrhosis and eighty patients with chronic hepatitis B were included in this study. The plasma amino acid profiles were measured by the amino acid analyzer. Patients were randomly divided into two groups, of which the liver cirrhosis group was to receive daily LES (n = 43) or non-LES (n = 43) for 6 months. Plasma amino acid profiles and biochemical parameters were measured in both groups at baseline and after 1, 3, and 6 months. RESULTS: Compared to healthy controls, the plasma concentration in the liver cirrhosis group of threonine, serine, glycine, glutamine, cysteine, tyrosine, phenylalanine, arginine, and methionine increased significantly (P < 0.05), while the ratio of branched chain amino acids (BCAA) to aromatic amino acids (AAA) decreased significantly (P < 0.05). A carbohydrate-predominant LES treatment resulted in a significant increase in BCAA/AAA and decrease in the level of ammonia and glutamine compared with baseline after 6 months of supplementation (P < 0.05). Patients with Child-Pugh B and C are more responsive to changes in amino acid profiles than those with Child-Pugh A. CONCLUSIONS: The application of an LES carbohydrate module for six months in liver cirrhosis patients was associated with increased BCAA/AAA and decreased level of ammonia. Patients with Child-Pugh B and C grades were the most beneficial population.

Characterization of metabolic landscape in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the most prevalent cancers worldwide, accounting for approximately 75%-85% of primary liver cancers. Metabolic alterations have been labeled as an emerging hallmark of tumors. Specially, the last decades have registered a significant improvement in our understanding of the role of metabolism in driving the carcinogenesis and progression of HCC. In this paper, we provide a review of recent studies that investigated the metabolic traits of HCC with a specific focus on three common metabolic alterations involving glycolysis, lipid metabolism, and glutamine addiction which have been gaining much attention in the field of HCC. Next, we describe some representative diagnostic markers or tools, and promising treatment agents that are proposed on the basis of the aforementioned metabolic alterations for HCC. Finally, we present some challenges and directions that may promisingly speed up the process of developing objective diagnostic markers and therapeutic options underlying HCC. Specifically, we recommend future investigations to carefully take into account the influence of heterogeneity, control for study-specific confounds, and invite the validation of existing biomarkers.

Blocking tumor necrosis factor-α delays progression of chronic obstructive pulmonary disease in rats through inhibiting MAPK signaling pathway and activating SOCS3/TRAF1.

The present study was conducted in order to study the detailed molecular mechanism of tumor necrosis factor (TNF)-α in chronic obstructive pulmonary disease (COPD). The rats were treated with cigarette smoke (CS) and lipopolysaccharide (LPS) to establish the COPD model. Next, the changes in lung injury in COPD rats with TNF-α knockdown was tested. Meanwhile, the regulation of TNF-α on MAPK pathway and its downstream molecules (SOCS3/TRAF1) was determined by western blotting. On this basis, the activation of MAPK and inhibition of SOCS3/TRAF1 was also examined. Subsequently, the lung function was tested with the plethysmograph, the cells of bronchoalveolar lavage fluid was counted and classified. Furthermore, lung tissue sections were stained with hematoxylin and eosin to verify whether the treatment of MAPK pathway and downstream molecules affected the effect of TNF-α knockdown on COPD. The present study showed that TNF-α knockdown could alleviate the decrease in the function and inflammatory injury of the lungs of rats with COPD. Western blot analysis verified that TNF-α knockdown could inhibit the activation of MAPK pathway and increase the expression of SOCS3/TRAF1. The following experimental results showed that the relief of lung injury caused by TNF-α knockdown could be deteriorated by activating MAPK pathway. It was also found that the symptom of COPD was decreased following transfection with sh-TNF-α but worsened by SOCS3/TRAF1 knockdown. Overall, TNF-α knockdown inhibited the activation of MAPK pathway and increased the expression of SOCS3/TRAF1, thus delaying the process of COPD.

Current understanding of mesenchymal stem cells in liver diseases.

Liver diseases caused by various factors have become a significant threat to public health worldwide. Liver transplantation has been considered as the only effective treatment for end-stage liver diseases; however, it is limited by the shortage of donor organs, postoperative complications, long-term immunosuppression, and high cost of treatment. Thus, it is not available for all patients. Recently, mesenchymal stem cells (MSCs) transplantation has been extensively explored for repairing hepatic injury in various liver diseases. MSCs are multipotent adult progenitor cells originated from the embryonic mesoderm, and can be found in mesenchymal tissues including the bone marrow, umbilical cord blood, adipose tissue, liver, lung, and others. Although the precise mechanisms of MSC transplantation remain mysterious, MSCs have been demonstrated to be able to prevent the progression of liver injury and improve liver function. MSCs can self-renew by dividing, migrating to injury sites and differentiating into multiple cell types including hepatocytes. Additionally, MSCs have immune-modulatory properties and release paracrine soluble factors. Indeed, the safety and effectiveness of MSC therapy for liver diseases have been demonstrated in animals. However, pre-clinical and clinical trials are largely required to confirm its safety and efficacy before large scale clinical application. In this review, we will explore the molecular mechanisms underlying therapeutic effects of MSCs on liver diseases. We also summarize clinical advances in MSC-based therapies.

Circ_0000388 Exerts Oncogenic Function in Cervical Cancer Cells by Regulating miR-337-3p/TCF12 Axis.

Background: Cervical cancer (CC) is one of the most common cancers among women worldwide. Circular RNAs (circRNAs) are recently identified as important gene regulators with critical roles in cancer biology. In this study, we explored the effects of circ_0000388 on the malignant phenotypes of CC cells and its mechanism. Materials and Methods: Circ_0000388 expression and miR-337-3p expression in CC tissue samples were measured using quantitative real time polymerase chain reaction. CCK-8 was adopted to assess the effect of circ_0000388 on CC cell line proliferation. TUNEL assay was employed to probe the effect of circ_0000388 on apoptosis. Wound healing assay and transwell assay were conducted to detect the effect of circ_0000388 on migration and invasion. Further, interaction among circ_0000388, miR-337-3p, and TCF12 (transcription factor 12) was determined by bioinformatics analysis, RT-PCR, Western blot, RNA immunoprecipitation assay, and luciferase reporter assay. Results: Circ_0000388 expression in CC clinical samples was upregulated and this was correlated with unfavorable pathological indexes. Circ_0000388 remarkably enhanced the proliferation and metastasis of CC cells. Circ_0000388 overexpression dramatically impeded miR-337-3p expression and it was identified as a sponge of miR-337-3p. Furthermore, circ_00003888 also enhanced the TCF12 expression, while the effect could be reversed by co-transfection with miR-377-3p. Conclusions: Circ_0000388 was a novel oncogenic circRNA in CC, and promoted cancer progression via regulating miR-337-3p and TCF12, and could be potentially used as a diagnostic biomarker and therapy target.

Epigallocatechin gallate from green tea exhibits potent anticancer effects in A-549 non-small lung cancer cells by inducing apoptosis, cell cycle arrest and inhibition of cell migration.

PURPOSE: Green tea (Camellia sinensis) is considered as a rich source of epigallocatechin gallate (EGCG) which has been shown to exert impressive pharmacological properties. The anticancer properties of EGCG have been extensively studied however, its anticancer activity has not been explored in lung cancer. The present study was therefore designed to evaluate the anticancer effects of EGCG against non-small cell lung cancer (NSCLC) cell line A-549 and normal human fibroblast FR-2 cells. METHODS: Cell viability was assessed by CCK8 assay, apoptosis by DAPI, annexin V/propidium iodide (PI) and flowcytometery and cell cycle analysis by flow cytometry. Cell migration capacity was investigated by wound-healing assay and protein expression was examined by Western blotting. RESULTS: The results revealed that EGCC could inhibit the proliferation of A-549 cells in a concentration-dependent manner and exhibited an IC50 of 25 µM against the IC50 of 100 µM against the normal human fibroblasts. Further evaluation revealed that EGCG exerts its anticancer effects via induction of apoptosis, modulation of Bax/blc-2 ratio and by triggering G2/M cell cycle arrest. Furthermore, EGCG could also inhibit the migration of A5-49 cells in a concentration-dependent manner. CONCLUSION: In conclusion, based on our results, we believe that EGCG could prove to be an important lead molecule for the treatment of lung cancer.

Evaluation of pluronic nanosuspensions loading a novel insoluble anticancer drug both in vitro and in vivo.

To improve the solubility, stability and the antitumor activity of a novel anticancer drug, 3-(4-bromopheny l)-2-(ethyl-sulfonyl)-6-methylquinoxaline1,4-dioxide (Q39), a poloxamer nanosuspension was developed by precipitation combined with high pressure homogenization in present study. In vitro characterizations of Q39 nanosuspension (Q39/NS), including particle size, polydispersity index (PI), morphology, crystalline, saturation solubility, stability and releases were evaluated. BABL/c nude mice bearing HepG2 cells were used as in vivo tumor models to evaluate the anti-tumor activity of Q39/NS after intravenous administration. The particle size and PI for Poloxamer188 nanosuspension (P188/NS) were (304±3) nm, and (0.123±0.005) respectively, and it was (307±5) nm and (0.120±0.007) for Poloxamer85 nanosuspension (P85/NS) correspondingly. The morphology of P188/NS was spherical shape while elliptoid shape for P85/NS. The crystalline of Q39/NS did not change as shown by the X-ray diffraction analysis. The stability of Q39/NS improved compared with the solution. The solubility of Q39 in P188/NS was 7.3 times higher than the original solubility, while it was 6 times for P85/NS. Sustained release as shown from the in vitro release test, together with the tumor-targeting as shown from in vivo NS distribution, may contribute to the enhanced in vivo antitumor activity of Q39/NS.

Serum level of adiponectin correlated with gender and genotype in patients with chronic hepatitis C.

Adiponectin is well recognized as plasma physiologically active polypeptide hormone exclusively derived from human and animal mature adipocytes, with vigorous property in antidiabetic, antiobesity, antiatherogenic, and anti-inflammatory processes. In this study, we investigated the correlation between serum adiponectin level and clinical and pathological parameters in patients with chronic hepatitis C (CHC). The study included 127 patients with CHC and 42 healthy volunteers as controls whose laboratory parameters and serum adiponectin and tumor necrosis factor-alpha (TNF-alpha) were assessed using enzyme-linked immunosorbent assay (ELISA). We demonstrated that a lower serum adiponectin level was associated with male gender, higher gamma-glutamyltransferase (gamma-GGT), higher albumin, higher TNF-alpha, and steatosis grade. The higher level of serum adiponectin in patients with genotype 2a was demonstrated when compared with that in the patients with genotype 1b. Furthermore, of great interest, results suggested that the significant differences regarding viral genotype seemed to occur only in male patients with CHC but not in female patients. In conclusion, serum adiponectin was associated with gender, genotype, liver steatosis, and TNF-alpha in a Chinese population with CHC.

[Relationship between the level of serum TNF-alpha of patients with chronic hepatitis C and treatment with interferon-alpha and the influencing factors].

OBJECTIVE: To assess the relationship between the level of serum TNF-alpha of chronic hepatitis C with severity of disease and curative effect of anti-virus therapy with interferon alpha. METHODS: Thirty healthy controls and 102 patients with chronic hepatitis C were recruited into this study. The level of serum TNF-alpha was determined by ELISA in both groups. Then the 102 patients with chronic hepatitis C were evaluated after being classified into mild (44 cases), moderate (34 cases), and severe types (24 cases) based on the results of liver function tests. Liver functions, viral load and genotype of HCV RNA were measured. RESULTS: Before anti-virus treatment, the level of fasting serum TNF-alpha in the patients with chronic hepatitis C (1) was higher than that of the normal controls; (2) in the cases with mild type was lower than that in the moderate and severe groups; (3) serum TNF-alpha was not associated with the HCV load; (4) serum TNF-alpha was not significantly different between HCV subtypes 1b and 2a; (5) serum TNF-alpha was not significantly different between the patients responsive and non-responsive to the anti-virus treatment; (6) serum TNF-alpha was positively correlated with the level of serum direct bilirubin, negatively correlated with cholinesterase (CHE). CONCLUSIONS: The level of fasting serum TNF-alpha in the patients with chronic hepatitis was higher than that in the normal controls, and was positively correlated with the severity of the disease but not correlated with the therapeutic effect of interferon-alpha.

[Effect of shenfu injection on brain natriuretic polypeptide and aminoterminal peptide of precollagen type III in patients with acute myocardial infarction during intervention treatment].

OBJECTIVE: To investigate the effect of Shenfu injection (SFI) on indexes of heart function and myocardial fibrosis in patients with acute myocardial infarction (AMI) treated by percutaneous coronary intervention (PCI). METHODS: Ninety-three AMI patients treated by PCI were randomly divided into two groups, 47 in the treatment group (treated by PCI plus 40 ml SFI intravenous injection, once a day for 7 days) and 46 in the control group (treated by PCI only). Levels of brain natriuretic poly peptide (BNP) and aminoterminal peptide of precollagen type III (NP III) were measured at different time points, i.e. immediately after admission (T1), and at 24th hour (T2) and 7th day (T3) after AMI onset in the two groups. RESULTS: The proportion of TIMI 3 grade of front blood flow in AMI related vessels after PCI was insignificantly different in the two groups. The concentration of BNP of T2 was significantly higher than that of T1 in the two groups (P < 0.01), and it was higher in the control group than that in the treatment group (P < 0.01), while at T3, it was insignificantly different in the treatment group to that of T1 (P > 0.05), but in the control group, it was still significantly higher than that in the treatment group and that of T1 (P < 0.01). The levels of NP III the two groups were similar at T1 and T2 (P > 0.05). At T3, it showed an increase in the treatment group, but with no significant difference (P > 0.05) as compared with that at T1, but in the control group did markedly increase and showed significant difference as compared with that of T1 and that in the treatment group at T3 (P < 0.05). CONCLUSION: Early applying of SFI can protect myocardium from ischemia/reperfusion injury after AMI, ameliorate the degree of injury, improve heart function of patients and prevent myocardial fibrosis.

[Relationship between p27 expression and prognosis of colorectal carcinoma].

BACKGROUND & OBJECTIVE: It has been reported that p27 is associated with prognosis of some tumor. This study was designed to investigate the relationship between expression of p27 and prognosis of colorectal carcinoma. METHODS: The expression of p27 in 82 colorectal carcinoma specimens was detected by immunohistochemical method. The relationship between the expression of p27 and clinicopathological characteristics or survival time was analyzed by chi 2 test or log rank test, respectively, while univariate and multivariate prognostic analyses were performed by using Cox regression model. RESULTS: High p27 expression rate in colorectal carcinoma was 53.66% (44/82), while low expression rate was 46.34% (38/82). Low p27 expression in colorectal carcinoma was significantly associated with late Dukes stage, high incidence of lymph metastasis, and distant metastasis. Long rank test showed that the p27 expression was positively correlated to survival time and Cox regression indicated that the p27 was an prognostic marker of colorectal carcinoma. CONCLUSION: Low p27 expression in colorectal carcinoma was significantly associated with late Dukes stage, high incidence of lymph metastasis, and distant metastasis, poor prognosis, and short survival time. p27 is an prognostic marker of colorectal carcinoma.