Application of herbs and active ingredients ameliorate non-alcoholic fatty liver disease under the guidance of traditional Chinese medicine.

Non-alcoholic fatty liver disease (NAFLD) is a global health problem, and its prevalence has been on the rise in recent years. Traditional Chinese Medicine (TCM) contains a wealth of therapeutic resources and has been in use for thousands of years regarding the prevention of liver disease and has been shown to be effective in the treatment of NAFLD in China. but the molecular mechanisms behind it have not been elucidated. In this article, we have updated and summarized the research and evidence concerning herbs and their active ingredients for the treatment in vivo and vitro models of NAFLD or NASH, by searching PubMed, Web of Science and SciFinder databases. In particular, we have found that most of the herbs and active ingredients reported so far have the effect of clearing heat and dispelling dampness, which is consistent with the concept of dampness-heat syndrome, in TCM theory. we have attempted to establish the TCM theory and modern pharmacological mechanisms links between herbs and monomers according to their TCM efficacy, experiment models, targets of modulation and amelioration of NAFLD pathology. Thus, we provide ideas and perspectives for further exploration of the pathogenesis of NAFLD and herbal therapy, helping to further the scientific connotation of TCM theories and promote the modernization of TCM.

Comprehensive analysis of nine m7G-related lncRNAs as prognosis factors in tumor immune microenvironment of hepatocellular carcinoma and experimental validation.

Background: Hepatocellular carcinoma (HCC) remains the most prevalent gastrointestinal malignancy worldwide, with robust drug resistance to therapy. N7-methylguanosine (m7G) mRNA modification has been significantly related to massive human diseases. Considering the effect of m7G-modified long non-coding RNAs (lncRNAs) in HCC progression is unknown, the study aims at investigating a prognostic signature to improve clinical outcomes for patients with HCC. Methods: Two independent databases (TCGA and ICGC) were used to analyze RNAseq data of HCC patients. First, co-expression analysis was applied to obtain the m7G-related lncRNAs. Moreover, consensus clustering analysis was employed to divide HCC patients into clusters. Then, using least absolute shrinkage and selection operator-Cox regression analysis, the m7G-related lncRNA prognostic signature (m7G-LPS) was first tested in the training set and then confirmed in both the testing and ICGC sets. The expression levels of the nine lncRNAs were further confirmed via real-time PCR in cell lines, principal component analysis, and receiver operating characteristic curve. The m7G-LPS could divide HCC patients into two different risk groups with the optimal risk score. Then, Kaplan-Meier curves, tumor mutation burden (TMB), therapeutic effects of chemotherapy agents, and expressions of immune checkpoints were performed to further enhance the availability of immunotherapeutic treatments for HCC patients. Results: A total of 1465 lncRNAs associated with the m7G genes were finally selected from the TCGA database, and through the univariate Cox regression, the expression levels of 22 m7G-related lncRNAs were concerning HCC patients' overall survival (OS). Then, the whole patients were grouped into two subgroups, and the OS in Cluster 1 was longer than that of patients in Cluster 2. Furthermore, nine prognostic m7G-related lncRNAs were identified to conduct the m7G-LPS, which were further verified. A prognostic nomogram combined age, gender, HCC grade, stage, and m7G-LPS showed strong reliability and accuracy in predicting OS in HCC patients. Finally, immune checkpoint expression, TMB, and several chemotherapy agents were remarkably associated with risk scores. More importantly, the OS of the TMB-high patients was the worst among the four groups. Conclusion: The prognostic model we established was validated by abundant algorithms, which provided a new perspective on HCC tumorigenesis and thus improved individualized treatments for patients.

Bufalin stimulates antitumor immune response by driving tumor-infiltrating macrophage toward M1 phenotype in hepatocellular carcinoma.

BACKGROUND: Immunotherapy for hepatocellular carcinoma (HCC) exhibits limited clinical efficacy due to immunosuppressive tumor microenvironment (TME). Tumor-infiltrating macrophages (TIMs) account for the major component in the TME, and the dominance of M2 phenotype over M1 phenotype in the TIMs plays the pivotal role in sustaining the immunosuppressive character. We thus investigate the effect of bufalin on promoting TIMs polarization toward M1 phenotype to improve HCC immunotherapy. METHODS: The impact of bufalin on evoking antitumor immune response was evaluated in the immunocompetent mouse HCC model. The expression profiling of macrophage-associated genes, surface markers and cytokines on bufalin treatment in vitro and in vivo were detected using flow cytometry, immunofluorescence, western blot analysis, ELISA and RT-qPCR. Cell signaling involved in M1 macrophage polarization was identified via the analysis of gene sequencing, and bufalin-governed target was explored by immunoprecipitation, western blot analysis and gain-and-loss of antitumor immune response. The combination of bufalin and antiprogrammed cell death protein 1 (anti-PD-1) antibody was also assessed in orthotopic HCC mouse model. RESULTS: In this study, we showed that bufalin can function as an antitumor immune modulator that governs the polarization of TIMs from tumor-promoting M2 toward tumor-inhibitory M1, which induces HCC suppression through the activation of effector T cell immune response. Mechanistically, bufalin inhibits overexpression of p50 nuclear factor kappa B (NF-κB) factor, leading to the predominance of p65-p50 heterodimers over p50 homodimers in the nuclei. The accumulation of p65-p50 heterodimers activates NF-κB signaling, which is responsible for the production of immunostimulatory cytokines, thus resulting in the activation of antitumor T cell immune response. Moreover, bufalin enhances the antitumor activity of anti-PD-1 antibody, and the combination exerts synergistic effect on HCC suppression. CONCLUSIONS: These data expound a novel antitumor mechanism of bufalin, and facilitate exploitation of a new potential macrophage-based HCC immunotherapeutic modality.

[Bushen Rougan Recipe in prevention of hepatic fibrosis in rats induced by dimethylnitrosamine: a study on its preliminary mechanism].

OBJECTIVE: To study the effects of Bushen Rougan Recipe (BSRGR), a compound traditional Chinese herbal medicine, on hepatic fibrosis in rats induced by dimethylnitrosamine (DMN), and to explore its preliminary mechanism. METHODS: A total of 40 male Wistar rats were randomly divided into normal control group (n=10), untreated group (n=15), and BSRGR group (n=15). Except for the rats in normal control group, hepatic fibrosis in rats was induced by peritoneal injection of DMN for 4 weeks. And the rats in the BSRGR group were also intragastrically administered BSRGR within the 4-week course. At the end of the 4-week course, rats were all sacrificed. The liver functions were determined by automatic biochemistry analyzer, including serum total bilirubin (Tbil), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and albumin. Expressions of connective tissue growth factor (CTGF) and collagen type I mRNA in liver tissue were detected by reverse transcription polymerase chain reaction. RESULTS: It was found that the serum Tbil level and the activities of AST, ALT were declined in the BSRGR group as compared with those in the untreated group (P<0.01). The serum albumin content in the BSRGR group was increased as compared with that in the untreated group (P<0.01). The expressions of collagen type I and CTGF mRNAs in the untreated group were higher than those in the BSRGR group (P<0.01). CONCLUSION: BSRGR can decrease the expressions of collagen type I and CTGF mRNAs in the rats with hepatic fibrosis, which may be one of possible mechanisms in treating hepatic fibrosis.

Bushen recipe and its disassembled prescriptions inhibit inflammation of liver injury associated with Concanavalin A through Toll­like receptor 3/9 signaling pathway.

The aim of the present study was to explore the effect of Bushen recipe and its disassembled prescriptions on liver injury and chronic hepatitis B. Liver injury was induced in normal and hepatitis B virus (HBV)­transgenic mice through injection of Concanavalin A, followed by treatment with Bushen recipe and its disassembled prescriptions including the Bushen­yang, the Bushen­yin and the QingHua groups as well as the GanYanLing group (positive control). Subsequently, their liver function indexes were investigated by a microplate method and liver sections were blindly evaluated using an optical microscope by a pathologist. Subsequently, the activation state of Toll­like receptor (TLR)3/9 signaling pathway in liver tissues was analyzed by western blotting. Additionally, the inflammatory factors produced following liver injury in peripheral blood were detected via ELISA. Following intervention with the Bushen recipe and its disassembled prescriptions, the liver function indexe alanine aminotransferase had declined, whereas cholinesterase increased. The pathological alterations of liver tissue in HBV transgenic mice were reversed by Bushen recipe and its disassembled prescriptions. In addition, the TLR3/9 signaling pathway in liver tissues of HBV transgenic mice was inhibited and inflammatory factors such as interleukin (IL)­6, IL­1, tumor necrosis factor­α and interferon­Î³ were reduced significantly. In conclusion, the present study demonstrated that Bushen recipe and its disassembled prescriptions repaired liver injury induced by Concanavalin A through inhibition of TLR3/9 signaling pathway.

[Effect of spleen-invigorating prescription on dendritic cell function in patients with chronic hepatitis B of TCM Pi-deficiency syndrome type].

OBJECTIVE: To observe the effect of Spleen-invigorating Prescription (SIP) on dendritic cell function in patients with chronic hepatitis B. METHODS: A total of 60 patients with chronic HBV of Pi-deficiency syndrome type were enrolled and randomized to 2 groups, 30 in each group. Patients in the control group were given intramuscular injection with human interferon alpha 1b, 3 times a week, while those in the treated group were given orally with SIP twice a day, the therapy lasted for 6 months. Dendritic cells (DCs) were isolated from peripheral blood and cultured, then the expression of surface markers, HLA-DR, CD86, CD80, CD40, CD14 and CD11c were detected before and after treatment by flow cytometry, and the function of DCs was also evaluated by mixed lymphocyte reaction (MLR) determination once before treatment and once after treatment. RESULTS: The expressions of DCs' surface CD86, CD80, CD40 and CD11c in the treated group were higher (P < 0.05, P < 0.01) and the changes of stimulating index, IFN-gamma and IL-12 were superior in the treated group to those in the control group (P < 0.05). CONCLUSIONS: SIP can significantly improve DC's function, so, one of mechanisms of SIP in improving clinical efficacy may be the regulation of immune function.

Prospects of a comprehensive evaluation system for traditional Chinese medicine services.

Traditional Chinese medicine (TCM) is a unique health resource in China and one of the main representative traditional medicines globally. TCM has formed a new way of looking at medical practices, health care, scientific research, education, industry and culture. It focuses on promoting and safeguarding the health of people, with an increasing contribution to economic and social development. Establishing a comprehensive evaluation system in accordance with the characteristics of TCM services could promote the scientific merit and the standardization of services management. This would improve health service quality and the social and economic benefits of TCM. It would broaden the field of TCM services research. It would also provide the basis for the formulation of relevant government policies. This study estimates the prospect of establishing a comprehensive evaluation system of TCM services.

[Clinical study on effect of bushen granule combined with marine injection in treating chronic hepatitis B of Gan-shen deficiency with damp-heat syndrome type].

OBJECTIVE: To evaluate the antiviral efficacy of Bushen Granule (BSG) combined with Marine Injection (MI) in treating patients with chronic hepatitis B of Gan-shen deficiency with Damp-Heat (GSD) syndrome type. METHODS: A total of 90 patients, who were HBV DNA, HBsAg, and HBeAg positive and of GS-DD type, were enrolled, and they were randomly assigned into 2 groups. The 49 patients in the treated group were treated with the combined therapy of BSG and MI and the 41 were administered with lamivudine, the therapeutic course for both groups was 1 year. The negative conversion rate of HBeAg and HBV DNA and the changes in HBV DNA titre, liver function, symptoms and physical signs were observed in the two groups before and after treatment. RESULTS: In the treated group, the negative conversion rate of HBV DNA was 42.6% (20/ 47), insignificantly different to that in the control group (61.0%, 25/41, P > 0.05); the negative conversion rate of HBeAg and HBeAg/anti-HBe sero-conversion rate was 42.6% and 36.2%, respectively, significantly higher than those in the control group (22.0% and 17.0%, P < 0.05); the ALT normalizing rate was 74.4%, higher than that in the control group (51.4%, P < 0.05); the improvement of liver function (ALB and GLB) and clinical symptoms, especially the dizziness, soreness of waist, hypochondrial distending pain, and yellowish urine, were significantly superior to those in the control group(P < 0.01 or P < 0.05); and the partial response rate was 29.8%, insignificant different to that in the control group (14.6%, P > 0.05). CONCLUSIONS: Combined treatment of BSG and MI has an ideal short-term effect in treating patients with chronic hepatitis B of Gan-shen deficiency with Damp-Heat syndrome type, it can inhibit HBV replication and improve patients' liver function and physical signs.

[Relationship between basal core promoter combined point mutation of hepatitis B virus and TCM syndrome type].

OBJECTIVE: To investigate the relationship between basal core promoter (BCP) combined point mutation of hepatitis B virus (HBV) and TCM syndrome type. METHODS: One hundred and two patients with chronic hepatitis with positive HBV DNA and hadn't ever been treated by Lamivudine and interferon were differentiated according TCM syndrome differentiation into 5 types, two excess types (damp-heat blocking zhong-jiao type and blood stasis blocking collaterals type) and three deficiency types, gan-stagnation with pi-dificiency type, gan-shen yin-deficiency type and pi-shen yang-deficiency type. The serum HBV DNA, hepatic biochemical indexes, and the mutation of BCPnt 1762A-T and nt1764G-A combined point were determined, respectively. RESULTS: The variant strain positive rate detected in the excess type was significantly higher than that in the deficiency type, the highest rate appeared in patients of damp-heat blocking zhong-jiao type. CONCLUSION: BCP combined point mutation may be liable to happen in patients of TCM excess type, especially in patients of damp-heat blocking zhong-jiao type.

[Epidemic survey on TCM syndrome typing and preliminary discussion on TCM etiology and pathogenesis of fatty liver].

OBJECTIVE: To investigate the common TCM syndrome types of fatty liver by way of epidemic questionnaire, their occurrence ratio, and the correlation between various syndrome types and objective indexes. METHODS: A total of national wide 503 subjects with fatty liver were enrolled, the TCM syndromes, body mass index (BMI), abdominal perimeter/hip circumference, liver function, blood lipids, B ultrasonic examination and CT in them were observed and recorded. RESULTS: In the 46 symptoms investigated in total, the first ten symptoms in order of appearing rate were lassitude, obese, oral dryness, vertigo, hypochondriac distending pain, soreness and pain in loin, spiritlessness, oral bitterness, aching and weakness in knee and abdominal distention. The mostly appeared tongue figures were pale and corpulent or pale dim tongue proper, white greasy or yellow greasy tongue coating, and the mostly appeared pulse figures were taut, taut-thin and taut slippery. Statistical cluster analysis showed that syndromes of fatty liver could be typed into 4 TCM types, the asthenia Pi-Shen with Gan-stagnation type, the asthenia Pi-Shen type, the asthenia Pi with phlegm-heat type and the unclassified type. Among them the asthenia Pi-Shen with Gan-stagnation type was the commonest one, which accounted to 62.32%. CONCLUSION: The mostly appeared syndrome type of fatty liver was asthenia Pi-Shen with Gan-stagnation type. The TCM pathogenesis of fatty liver was deficiency of origin, mainly deficiency of Shen, involving Pi, with excess superficiality, the turbid-phlegm and blood stasis.

[Therapeutic effect of Bushen Rougan Recipe on hepatic fibrosis in rats].

OBJECTIVE: To study the therapeutic effect of Bushen Rougan Recipe (BSRGR) on hepatic fibrosis in rats. METHODS: Forty male Wistar rats were randomly divided into normal control group (n=10), model group (n=15), and BSRGR-treated group (n=15). Rats in the model and BSRGR-treated groups were administered intraperitoneally with 0.5% dimethylnitrosamine (DMN), 10 mg.kg(-1).d(-1), successive 3 days per week for 4 weeks to induce hepatic fibrosis. Then rats in the above 2 groups were given normal saline and BSRGR (10 ml.kg(-1).d(-1), ig) for another 4 weeks, respectively. Rats in the 3 groups were all executed at the end of the 8th week. The serum total bilirubin (TBIL), alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (A) and globulin (G) were determined and the serum hyaluronic acid (HA), laminin (LN) and collagen IV (IV-C) were measured. RESULTS: The rat model of liver fibrosis was successfully induced by DMN. It was found that the serum TBIL, AST and ALT and the liver fibrosis marks were declined in BSRGR-treated group as compared with those in the model group (P<0.01). The content of total serum protein and the A/G in BSRGR-treated group were both increased as compared with those in the model group (P<0.05). CONCLUSION: BSRGR can be used to treat hepatic fibrosis in rats.

Chinese medicine bu xu hua yu recipe for the regulation of treg/th17 ratio imbalance in autoimmune hepatitis.

Objectives. The aim of this study is researching the role of the Regulatory T cell (Treg)/T helper cell-17 (Th17) cell ratio imbalance in the pathogenesis of autoimmune hepatitis (AIH) and the use of the "Bu Xu Hua Yu" recipe in the treatment of AIH. Materials and Methods. Sixty adult male C57/BL6 mice were divided into six different groups. α-Galcer was injected abdominally for production of the animal models. Liver function tests, histological examinations, liver tissue Regulatory T cell, and T helper cell-17 levels tests were carried out. TGF-ß1, IL-10, IL-17, and expression of mRNA and protein levels of Foxp3 and ROR-γt were also assessed. Results. Bu Xu Hua Yu method increased the levels of Regulatory T cell, IL-10, and the expression of Foxp3 (P < 0.05) in mice liver tissues. Furthermore, there were decreases in the levels of T helper cell-17, IL-17, and expression of RORγt mRNA and protein (P < 0.05). The ratio of Treg/Th17 was increased (P < 0.05). Conclusion. Mice with AIH have a Treg/Th17 ratio imbalance. Bu Xu Hua Yu method was able to restore the cellular balance of Treg/Th17 through the regulation of the expression of RORγt and Foxp3 and can play an important role in the treatment of AIH.

Liver stiffness measurement, better than APRI, Fibroindex, Fib-4, and NBI gastroscopy, predicts portal hypertension in patients with cirrhosis.

Liver stiffness measurement (LSM) is frequently used as non-invasive alternative for liver fibrosis including cirrhosis, which can lead to portal hypertension. This study was conducted to evaluate the predictive value of LSM in cirrhosis-induced portal hypertension patients. Between July 2011 and December 2013, 153 participants were enrolled into a single-center, prospective, cross-sectional study. Each subject received both gastroscopy and LSM. Baseline biochemical, APRI, Fibroindex, and Fib-4 were also performed. LSM of cirrhosis patients with portal hypertension was significantly higher compared to those without portal hypertension (P < 0.05). A LSM ≥ 13.6 kPa had a sensitivity of 83.87 % and a specificity of 72.53 % with an accuracy of 77.1 for the prediction of portal hypertension, which are higher than those of APRI, Fib-4, and Fibroscan separately. A combination of Fibroscan combined with Fib-4 achieved a maximum AUC of 0.833 and accuracy of 77.8. Discriminant and internal validation analysis showed that Fibroscan plus APRI obtained a lower false negative rate compared to Fibroscan plus Fib-4 and Fibroscan plus Fibroindex (9.68, 17.74, and 11.29 %, respectively). A good relationship was found between LSM and NBI mean optical density both by linear and polynomial correlation analysis (r = 0.5533 and r = 0.7349, both P < 0.001). There was a trend toward a better performance of LSM for assessing portal hypertension compared with NBI gastroscopy mean optical density (P = 0.028 and P = 0.05, respectively). Better than APRI, Fibroindex, Fib-4, and NBI gastroscopy, LSM can predict portal hypertension in cirrhosis patients. A LSM of 13.6 kPa can be considered to be the predictive value for portal hypertension.

[Effect of anti-fibrosis compound contained serum on procollagen Type I and IV, matrix metalloproteinase and its tissue inhibitor-1 gene expression in HSC-LI90 cell line].

OBJECTIVE: To study the effects of Anti-fibrosis Compound contained serum (AFCS) on procollagen type I and IV (ProC-I and ProC-IV), matrix metalloproteinase (MMP) and its tissue inhibitor (TIMP-1) gene expression in hepatic stellate cell line LI90 (HSC-LI90). METHODS: AFCS was prepared by gastric infusing different dosage (0.5 g/kg, 2.0 g/kg and 4.0 g/kg) of Anti-fibrosis Compound Recipe to rats. After HSC-LI90 cells were exposed to AFCS for 48 hrs, levels of ProC-I, ProC-IV, gene expression of MMP-2, MMP membrane type 1 (MT1-MMP) and TIMP-1 in the cells were detected by Northern blot, and gelatinase activity of MMP-2 was measured by zymography. RESULTS: AFCS of different concentrations could inhibit ProC-I and ProC-IV and TIMP-1 gene expression (P < 0.05 or P < 0.01), increase MT1-MMP gene expression (P < 0.01), but it showed no effect on gene expression and activity of MMP-2 (P > 0.05). CONCLUSION: Anti-fibrosis Compound Recipe has anti-liver fibrosis action, its effects in inhibiting TIMP-1 gene expression of HSC-LI90 cells and promoting degradation of collagen might be one of the mechanisms of the action.

[Therapeutic efficacy of combined application of lamivudine and bushen recipe in treating chronic hepatitis B and its influence on YMDD motif].

OBJECTIVE: To observe the effect of combined application of Lamivudine and Bushen recipe (BSR) in treating patients with chronic hepatitis B (CHB) and its influence on tyrosine-methionine-aspartate-aspartate (YMDD) motif. METHODS: Eighty-eight patients of CHB with positive HBV-DNA were divided into 3 groups, Group A treated with Lamivudine, Group B treated with Lamivudine plus BSR1 and Group C treated with Lamivudine plus BSR2, the serum HBV-DNA, hepatic biological parameters and YMDD motif mutation were observed. RESULTS: The positive rate of HBV-DNA (6.45%) and YMDD motif mutation (3.23%) in Group C were lower than those in Group A respectively, after 52 weeks treatment. CONCLUSION: Combined application of Lamivudine and BSR could enhance the therapeutic effect in treating CHB to certain extent, and could reduce the mutation of YMDD motif.