Integrating proteomics and targeted metabolomics to reveal the material basis of liver-gallbladder damp-heat syndrome in chronic hepatitis B

Objective To elucidate the biological basis of liver-gallbladder damp-heat syndrome(LGDHS)within the framework of traditional Chinese medicine(TCM)as a complementary diagnostic and therapeutic approach in chronic hepatitis B(CHB).Methods CHB patients and healthy volunteers were enrolled from Shuguang Hospital Affili-ated to Shanghai University of Traditional Chinese Medicine between August 21,2018 and December 31,2020.They were divided into three groups:healthy group,LGDHS group,and latent syndrome(LP)group.Proteomic analysis using isobaric tags for relative and absolute quantitation(iTRAQ)was performed to identify differentially expressed proteins(DEPs).Metabolomic profiling via ultra-performance liquid chromatography-tandem mass spec-trometry(UPLC-MS/MS)was applied to serum samples to detect differentially regulated metabolites(DMs).Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology(GO)enrichment were employed to explore dysregulated pathways.Principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)were utilized to visualize group separation and identify key metabolites and proteins contributing to LGDHS differentiation.Receiver operating characteristic(ROC)curve analysis evaluated the diagnostic performance of key biomarkers,while logistic regression models assessed their predictive accuracy.P values were corrected for multiple tests using the Benjamini-Hochberg method to control the false discovery rate(FDR).Validation of potential biomarkers was con-ducted using independent microarray data and real-time quantitative polymerase chain reac-tion(RT-qPCR).Results A total of 150 participants were enrolled,including healthy group(n=45),LGDHS group(n=60),and LP group(n=45).254 DEPs from proteomics data and 72 DMs from metabolomic profiling were identified by PCA and OPLS-DA.DEPs were mainly enriched in immune and complement pathways,while DMs involved in amino acid and energy metabolism.The integrated analysis identified seven key biomarkers:α1-acid glycoprotein(ORM1),asparagine synthetase(ASNS),solute carrier family 27 member 5(SLC27A5),glu-cosidase II alpha subunit(GANAB),hexokinase 2(HK2),5-methyltetrahydrofolate-homocys-teine methyltransferase(MTR),and maltase-glucoamylase(MGAM).Microarray validation confirmed the diagnostic potential of these genes,with area under the curve(AUC)values for ROC analysis ranging from 0.536 to 0.759.Among these,ORM1,ASNS,and SLC27A5 showed significant differential ability in differentiating LGDHS patients(P=0.016,P=0.035,and P<0.001,respectively),with corresponding AUC of 0.749,0.743,and 0.759,respectively.A logis-tic regression model incorporating these three genes demonstrated an AUC of 0.939,indicat-ing a high discriminatory power for LGDHS.RT-qPCR further validated the differential ex-pression of ORM1 and SLC27A5 between LGDHS and LP groups(P=0.011 and P=0.034,re-spectively),with ASNS showing a consistent trend in expression(P=0.928).Conclusion This study integrates multi-omics approaches to uncover the molecular mecha-nisms underlying LGDHS in CHB.The identification of biomarkers ORM1,ASNS,and SLC27A5 offers a solid basis for the objective diagnosis of LGDHS,contributing to the stan-dardization and modernization of TCM diagnostic practices.

Effect of JIANPI HUOXUE decoction on inflammatory cytokine secretion pathway in rat liver with lipopolysaccharide challenge

AIM： To evaluate the effect of Chinese traditional medicinal prescription, JIANPI HUOXUE decoction （JHD） on cytokine secretion pathway in rat liver induced by lipopolysaccharide （LPS）. METHODS： Twenty-four male SD rats were divided into normal group （n = 4）, model group （n = 10） and JHD group （n = 10） randomly. Rats in model group and JHD group were administrated with normal saline or JHD via gastrogavage respectively twice a day for 3 d. One hour after the last administration, rats were injected with LPS via tail vein, 50 μg/kg. Simultaneously, rats in normal group were injected with equivalent normal saline. After LPS stimulation for 1.5 h, serum and liver tissue were collected. Pathological change of liver tissues was observed through hematoxylineosin （H.E.） staining. Tumor necrosis factor alpha （TNF-α） in serum were assayed by enzyme linked immunosorbent assay （ELISA）. The protein expression of TNF-α, phosphorylated inhibit-κB （p-κB） and CD68 in liver were assayed by Western blot. The distribution of CD68 protein in liver was observed through immunohistochemical staining. The mRNA expression of TNF-α, interleukin-6 （IL-6）, CD14, toll-like receptor 2 （TLR2） and TLR4 in liver were assayed by real-time RT-PCR.RESULTS： Predominant microvesicular change, hepatocyte tumefaction and cytoplasm dilution were observed in liver tissues after LPS administration as well as obvious CD68 positive staining in hepatic sinusoidal. After LPS stimulation, serum TNF-α （31.35 ± 6.06 vs 12225.40 ± 9007.03, P 〈 0.05）, protein expression of CD68 （1.13 ± 0.49 vs 3.36 ±1.69, P 〈 0.05）, p-IκB （0.01 ±0.01 vs 2.07 ＋ 0.83, P 〈 0.01） and TNF-α （0.27 ± 0.13 vs 1.29 ± 0.37, P 〈 0.01） in liver and mRNA expression of TNF-α （1.96 ± 2.23 vs 21.45 ±6.00, P 〈 0.01）, IL-6 （4.80 ± 6.42 vs 193.50 ± 36.36, P 〈 0.01） and TLR2 （1.44 ± 0.62 vs 4.16 ± 0.08, P 〈 0.01） in liver were also increased significantly. These pathological changes were all improved in .1HD group. On the other hand, TLR4 mRNA （1.22 ± 0.30 vs 0.50 ± 0.15, P 〈 0.05） was down-regulated and CD14 mRNA increased but not significantly after LPS stimulation. CONCLUSION： JHD can inhibit cytokine secretion pathway induced by LPS in rat liver, which is probably associated with its regulation on CD68, p-IκB and endotoxin receptor TLR2.

Urinary metabolomics analysis identifies key biomarkers of different stages of nonalcoholic fatty liver disease

AIM To identify a panel of biomarkers that can distinguish between non-alcoholic fatty liver disease(NAFLD) and non-alcoholic steatohepatitis(NASH), and explore molecular mechanism involved in the process of developing NASH from NAFLD.METHODS Biomarkers may differ during stages of NAFLD. Urine and blood were obtained from non-diabetic subjects with NAFLD and steatosis, with normal liver function(n = 33), from patients with NASH, with abnormal liver function(n = 45), and from healthy age and sex-matched controls(n = 30). Samples were subjected to metabolomic analysis to identify potential non-invasive biomarkers. Differences in urinary metabolic profiles were analyzed using liquid chromatography tandem mass spectrometry with principal component analysis and partial least squares-discriminate analysis.RESULTS Compared with NAFLD patients, patients with NASH had abnormal liver function and high serum lipid concentrations. Urinary metabonomics found differences in 31 metabolites between these two groups, including differences in nucleic acids and amino acids. Pathway analysis based on overlapping metabolites showed that pathways of energy and amino acid metabolism, as well as the pentose phosphate pathway, were closely associated with pathological processes in NAFLD and NASH.CONCLUSION These findings suggested that a panel of biomarkers could distinguish between NAFLD and NASH, and could help to determine the molecular mechanism involved in the process of developing NASH from NAFLD. Urinary biomarkers may be diagnostic in these patients and could be used to assess responses to therapeutic interventions.

Therapeutic mechanism of Yīn-Chén-Hāo decoction in hepatic diseases

Yīn-Chén-Hāo decoction(YCHD) is a traditional Chinese medicine formula composed of capillaris(Artemisia capillaris), gardenia(Gardenia jasminoides), and rhubarb(Rheum rhabarbarum) that is used for the treatment of damp-heat jaundice. In modern clinics, YCHD is mostly used for hepatic diseases. This review summarizes the biological activities of YCHD and its medical applications. The main active compounds of YCHD are chlorogenic acid, rhein, geniposide, emodin, and scoparone. The pharmacological actions of YCHD include inhibition of hepatic steatosis, apoptosis, necrosis, anti-inflammation, and immune regulation. YCHD could be developed as a new therapeutic strategy for the treatment of hepatic diseases.

Synergistic effect of a novel oxymatrine-baicalin combination against hepatitis B virus replication, a smooth muscle actin expression and typeⅠcollagen synthesis in vitro

AIM： To study the effect of oxymatrine-baicalin combination （OB） against HBV replication in 2.2.15 cells and α smooth muscle actin （ α SMA） expression, type I, collagen synthesis in HSC-T6 cells. METHODS： The 2.2.15 cells and HSC-T6 cells were cultured and treated respectively. HBsAg and HBeAg in the culture supernatants were detected by ELISA and HBV DNA levels were determined by fluorescence quantitative PCR. Total RNA was extracted from HSC-T6 cells and reverse transcribed into cDNA. The cDNAs were amplified by PCR and the quantities were expressed in proportion to β actin. The total cellular proteins extracted from HSC-T6 cells were separated by electrophoresis. Resolved proteins were electrophoretically transferred to nitrocellulose membrane. Protein bands were revealed and the quantities were corrected by β actin. RESULTS： In the 2.2.15 cell culture system, the inhibitory rate against secretion of HBsAg and HBeAg in the OB group was significantly stronger than that in the oxymatrine group （HBsAg, P = 0.043; HBeAg, P = 0.026; respectively）; HBV DNA level in the OB group was significantly lower than that in the oxymatrine group （P = 0.041）. In HSC-T6 cells the mRNA and protein expression levels of α SMA in the OB group were significantly lower as compared with those in the oxymatrine group （mRNA, P = 0.013; protein, P = 0.042; respectively）; The mRNA and protein expression levels of type I collagen in the OB group were significantly lower as compared with those in the oxymatrine group （mRNA, P 〈 0.01; protein, P 〈 0.01; respectively）.CONCLUSION： OB combination has a better effect against HBV replication in 2.2.15 cells and is more effective against α SMA expression and type I collagen synthesis in HSC-T6 cells than oxymatrine in vitro.

Impact of Fuzheng Huayu tablet on antiviral effect of entecavir in patients with hepatitis B cirrhosis

Background:Fuzheng Huayu tablet is a traditional Chinese medicine(TCM)used for the treatment of liver fibrosis and cirrhosis.However,whether the combination with Fuzheng Huayu tablet could affect the antiviral efflcacy of nucleos(t)ide remains a concern.The objective of this trial was to explore the impact of Fuzheng Huayu tablet on antiviral effect of entecavir in patients with hepatitis B cirrhosis.Methods:A prospective,randomized control trial was conducted.Patients with compensated hepatitis B cirrhosis were randomly divided into the treatment group(entecavir capsule plus Fuzheng Huayu tablet)and the control group(entecavir capsule plus simulant of Fuzheng Huayu),and followed up for 48 weeks.The dynamic changes of HBV DNA load,the rate of serological conversion of HBeAg,liver function,renal function and liver stiffness measurement(LSM)were monitored.The general clinical data and adverse events were also recorded.Results:There was no significant difference in the rate of virological response and cumulative virological response between the treatment group and the control group(P>0.05).After 48 weeks of treatment,the HBeAg seroconversion rate,biochemical response rate and LSM value were 21.05%and 4.76%(P=0.164),86.96%and 65.96%(P=0.017),9.5 kpa and 10.6 kpa(P=0.827)in the treatment group and the control group,respectively.No serious adverse events related to the study therapy occurred during the trial.Conclusions:The antiviral entecavir combined with Fuzheng Huayu tablet did not affect the antiviral efflcacy of entecavir,but could improve the rate of biochemical response,and had a tendency to improve the rate of serological conversion of HBeAg and liver fibrosis in patients with hepatitis B cirrhosis.Fuzheng Huayu tablet is clinically safe for patients with hepatitis B cirrhosis.

Concomitant adenosquamous carcinoma and cystadenocarcinoma of the extrahepatic bile duct: A case report

BACKGROUND Infiltrative adenosquamous carcinoma(ASC) of the extrahepatic bile duct is reported infrequently, which is an unusual variant of the ordinary adenocarcinoma. The simultaneous development of ASC and cystadenocarcinoma in the extrahepatic biliary tree is rare. In addition, the accurate preoperative diagnosis of concomitant carcinoma in the multiple biliary trees at an early stage is often difficult. Thus, awareness of the risk of the multiplicity of biliary tumors is perhaps the most important factor in identifying these cases.CASE SUMMARY Here, we report a case of a 63-year-old female with jaundice, who was referred to Shuguang Hospital because of abdominal pain for 1 mo. An abdominal contrastenhanced computed tomography revealed a type I choledochal cyst and intraluminal masses suggestive of adenoma of the common bile duct. In addition,a preoperative diagnosis of a concomitant Klatskin tumor and type I choledochal cyst was made. The patient underwent anti-inflammatory therapy, followed by radical surgery due to hilar cholangiocarcinoma and resection of the choledochal cyst. Examination of the surgical specimen revealed a papillary tumor of the common bile duct, which arose from the malignant transformation of a preexisting cystadenoma. Histologic examination confirmed a special type of cholangiocarcinoma; the tumor in the hilar bile duct was an ASC, whereas the tumor in the common bile duct was a moderately differentiated cystadenocarcinoma. The patient showed rapid deterioration 8 mo after surgery.CONCLUSION Although concomitant ASC and cystadenocarcinoma of the extrahepatic bile duct is difficult to diagnose before surgery, and the prognosis is poor after surgery,surgical resection is still the preferred treatment.

Histological efficacy of traditional Chinese medicine plus entecavir therapy for HBV-related liver fibrosis

Background/Aims:Traditional Chinese medicine(TCM)combined with antiviral therapy has been proven to be effective for liver fibrosis due to chronic hepatitis B(CHB)in clinical practice in China.However,the robust evidence is limited,and the validity of results has been controversial in the past.The current study is to evaluate the efficacy and safety of the combination therapy of TCM plus entecavir(ETV)in the management of HBV-associated liver fibrosis or cirrhosis.Methods:Seven electronic databases were searched from inception to 10 August 2021.Primary outcome of this study was the regression of liver fibrosis;the secondary outcomes were a necro-inflammatory improvement,alanine aminotransferase(ALT),HBV DNA undetectable rate,HBeAg loss and HBeAg seroconversion.All the trials included were assessed by the Cochrane risk-of-bias tool.Results:It showed that TCM plus ETV attenuated liver fibrosis or cirrhosis in chronic hepatitis B patients as compared to ETV monotherapy(OR=1.65;95%CI:1.29~2.11;P<0.000,1).There is no statistical difference between TCM plus ETV and ETV in histological activity index,HBV DNA undetectable rate,HBeAg loss,HBeAg seroconversion and adverse events(P<0.05).Conclusion:The comprehensive evaluation showed that TCM combined with ETV treatment was safe for the patients with CHB,and better promoted the regression of liver fibrosis than ETV monotherapy.However,the standardized,rigorously designed,and large-scale randomized controlled trials(RCTs)were needed for further validation.

Mitochondria at the Crossroads of Cholestatic Liver Injury:Targeting Novel Therapeutic Avenues

Bile acids are byproducts of cholesterol metabolism in the liver and constitute the primary components of bile.Disruption of bile flow leads to cholestasis,characterized by the accumulation of hydrophobic bile acids in the liver and bloodstream.Such accumulation can exacerbate liver impairment.This review discussed recent developments in understanding how bile acids contribute to liver damage,including disturbances in mitochondrial function,endoplasmic reticulum stress,inflammation,and autophagy dysfunction.Mitochondria play a pivotal role in cholestatic liver injury by influencing hepatocyte apoptosis and inflammation.Recent findings linking bile acids to liver damage highlight new potential treatment targets for cholestatic liver injury.

Altered oral microbiota in chronic hepatitis B patients with different tongue coatings

AIM To elucidate tongue coating microbiota and metabolic differences in chronic hepatitis B(CHB) patients with yellow or white tongue coatings.METHODS Tongue coating samples were collected from 53 CHBpatients(28 CHB yellow tongue coating patients and 25 CHB white tongue coating patients) and 22 healthy controls.Microbial DNA was extracted from the tongue samples,and the bacterial 16 S ribosomal RNA gene V3 region was amplified from all samples and sequenced with the Ion Torrent PGM^(TM)sequencing platform according to the standard protocols.The metabolites in the tongue coatings were evaluated using a liquid chromatographymass spectrometry(LC-MS) platform.Statistical analyses were then performed.RESULTS The relative compositions of the tongue coating microbiotas and metabolites in the CHB patients were significantly different from those of the healthy controls,but the tongue coating microbiota abundances and diversity levels were not significantly different.Compared with the CHB white tongue coating patients,the CHB yellow tongue coating patients had higher hepatitis B viral DNA(HBV-DNA) titers(median 21210 vs 500,respectively,P = 0.03) and a significantly lower level of Bacteroidetes(20.14% vs 27.93%,respectively,P = 0.013) and higher level of Proteobacteria(25.99% vs 18.17%,respectively,P = 0.045) in the microbial compositions at the phylum level.The inferred metagenomic pathways enriched in the CHB yellow tongue coating patients were mainly those involved in amino acid metabolism,which was consistent with the metabolic disorder.The abundances of bacteria from Bacteroidales at the order level were higher in the CHB white tongue coating patients(19.2% vs 27.22%,respectively,P = 0.011),whereas Neisseriales were enriched in the yellow tongue coating patients(21.85% vs 13.83%,respectively,P = 0.029).At the family level,the abundance of Neisseriaceae in the yellow tongue patients was positively correlated with the HBV-DNA level but negatively correlated with the S-adenosyl-L-methionine level.CONCLUSION This research illustrates specific clinical features and bacterial structures in CHB patients with different tongue coatings,which facilitates understanding of the traditional tongue diagnosis.

Structural Shifts of Gut Flora in Rat Acute Alcoholic Liver Injury and Jianpi Huoxue Decoction's(健脾活血汤)Effect Displayed by ERIC-PCR Fingerprint

Objective： To study the structural shifts of gut flora in rats with acute alcoholic liver injury （AALI）, and the effect of Jianpi Huoxue Decoction （健脾活血汤, JPHXD） on the gut flora. Methods： Thirty-six Sprague- Dawley rats were randomly allocated to the control, AALI and JPHXD groups equally. The rats in the control group were given water and those in AALI and JPHXD groups were given ethanol by intragastric gavage for 5 days, while rats in the JPHXD group were administered JPHXD simultaneously. The blood and liver tissue were collected at the end of the experiment. The activities of serum alkaline aminotransferase （ALT）, aspartate aminotransferase （AST）, hepatic γ/-glutamyitranspetidase （γ-GT） and hepatic tdglyceride （TG） levels were determined. Plasma endotoxin level in the portal vein was measured. Pathological changes of liver tissues were determined by hematoxylin and Eosin （HE） staining and oil red O staining. The total DNA of gut flora were extracted from fecal samples by Bead-beating method and determined by ERIC-PCR fingerprint method. The similarity cluster analysis and principal component analysis were performed to analyze the ERIC-PCR fingerprint respectively. Results： In the AALI group, the ratio of liver/body weight, activities of ALT, AST and hepatic γ-GT, amount of hepatic TG were elevated significantly compared with those in the control group （all P〈0.01）. JPHXD decreased the ratio, activities of ALT, AST, γ-GT and TG significantly compared with those in the AALI group （P〈0.05 or P〈0.01）. HE and oil red O staining showed that fat deposited markedly in liver tissue, while JPHXD alleviated pathological changes markedly. Plasma LPS level in rat portal vein in the AALI group increased significantly （P〈0.01）, but it was decreased significantly in the JPHXD group （P〈0.01）. The cluster analysis and principal component analysis of ERIC-PCR fingerprint showed that gut flora in the hALl group changed markedly, and JPHXD could recover gut flora to some extent. Conclusion： The structure of gut flora shifted markedly during acute alcoholic liver injury, JPHXD had prevention effect through the modification of gut flora.

Xiayuxue Decoction(下瘀血汤)Attenuates Hepatic Stellate Cell Activation and Sinusoidal Endothelium Defenestration in CCI_4-Induced Fibrotic Liver of Mice

Objective： To investigate the effects of ancient Chinese medical formula Xiayuxue Decoction （下瘀血汤, XYXD） on activation of hepatic stellate cells （HSCs） and defenestration of sinusoidal endothelial cells （SECs） in CCI4-induced fibrotic liver of mice. Methods： High performance liquid chromatography was used to identify the main components of XYXD and control the quality of extraction. C57BL/6 mice were induced liver fibrosis by CCI4 exposure and administered with XYXD for 6 weeks simultaneously. Liver tissue was investigated by hematoxylin-eosin and Sirius-red staining. Sinusoidal fenestrations were observed by scanning electronic microscopy and fluorescent immunohistochemistry of PECAM-1 （CD31）. Whole liver lysates were detected of α-smooth muscle actin （α-SMA） and type-I collagen by Western blot. Primary rat HSCs-T6 cells were analyzed by detecting α-SMA, F-actin, DNA fragmentation through confocal microscopy, Western blot, terminal-deoxynucleoitidyl transferase mediated nick end labeling （TUNEL） assay and cellomics arrayscan, respectively. Results： Amygdalin and emodin in XYXD were identified. XYXD （993 mg/kg） inhibited Sirius red positive area up to 70.1% （P〈0.01）, as well as protein levels of α-SMA and type- I collagen by 42.0% and 18.5% （P〈0.05） respectively. In vitro, XYXD （12.5 μg/mL, 50 μg/mL） suppressed the activation of HSCs and reversed the myofibroblastic HSCs into quiescent, demonstrated as inhibition of fluorescent F-actin by 32.3% and 46.6% （P〈0.05）. Besides, XYXD induced the apoptosis of HSC-T6 cells by 20.0% （P〈0.05） and 49.5% （P〈0.01）, evidenced by enhanced TUNEL positivity. Moreover, ultrastructural observation suggested XYXD inhibited defenestration of SECs, which was confirmed by 31.1% reduction of protein level of CD31 （P〈0.05）. Conclusions： XYXD inhibited both HSCs activation and SECs defenestration which accompany chronic liver injuries. These data may help to understand the underlying mechanisms of XYXD for prevetion of chronic liver diseases.

Intervention Effects of Jianpi Liqi Huoxue Decoction (健脾理气活血汤) on Lipid Peroxidative Liver Injury Induced by Alcohol

Objective： To study the intervention effects of Jianpi Liqi Huoxue Decoction （健脾理气活血汤, JLHD） on lipid peroxidative liver injury induced by alcohol. Methods： The rat alcoholic model of liver disease （ALD） induced by Lieber-DeCarli liquid diet was established. Thirty-two male SD rats were randomly divided into 4 groups ： the normal group （ n = 5）, the control group （ n = 9）, the model group （ n = 9） and the JLHD group （ n =9）. From the 4th week after modeling, the rats were given JLHD or distilled water by gastrogavage respectively, and the samples of blood and liver tissues were taken out from the rats for determination by the end of the 8th week. The hepatic pathological changes were observed with HE staining; the liver injury related indices, including activity of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum, γ-glutamyl transpeptidase （γ-GT） activity and triglyceride （TG） content in liver tissues, as well as the lipid peroxidation related indices, including malonaldehyde （MDA） content and nitric oxide synthase （NOS） activity in liver tissue, serum Fe^2＋ level, and the anti-peroxidation capacity related indices, including superoxide dismutase （SOD） activity, glutathion （GSH） content and reactive oxygen species （anti- ROS） activity in liver tissues were determined. Results： （ 1 ） There were obvious figures of fatty degeneration and inflammatory infiltration in liver tissues of the model group. As compared with the control group, in the model group, the activity of ALT and AST, and Fe^2＋ content in serum, γ-GT and NOS activity, TG and MDA content in liver tissues were significantly higher （ P〈0. 01 ）, while the activity of SOD, GSH and anti-ROS in liver tissues were significantly lower （P〈0.01）. （2） The fatty degeneration and inflammatory infiltration of liver tissues in the JLHD group were significantly lessen as compared with those in the model group; and the abnormalities of all the indexes revealed in the model rats were restored to certain extent in the JLHD group, and especially significant were the levels of ALT activity, MDA content and Fe^2＋ , which were nearly normal. Conclusion： JLHD has significant effects against alcoholic liver injury, the acting mechanism of which is likely to be related with its anti-lipid peroxidative effect.

Research progress on signaling pathways in cirrhotic portal hypertension

Portal hypertension(PHT) is an important consequence of liver cirrhosis, which can lead to complications that adversely affect a patient's quality of life and survival, such as upper gastrointestinal bleeding, ascites, and portosystemic encephalopathy. In recent years, advances in molecular biology have led to major discoveries in the pathological processes of PHT, including the signaling pathways that may be involved: PI3 K-AKT-mTOR, RhoA/Rho-kinase, JAK2/STAT3, and farnesoid X receptor. However, the pathogenesis of PHT is complex and there are numerous pathways involved. Therefore, the targeting of signaling pathways for medical management is lagging. This article summarizes the progress that has been made in understanding the signaling pathways in PHT, and provides ideas for treatment of the disorder.

Intestinal Microecology:An Important Target for Chinese Medicine Treatment of Non-alcoholic Fatty Liver Disease

Non-alcoholic fatty liver disease(NAFLD)has become one of the most common chronic liver diseases worldwide,causing serious economic and medical burdens.Currently,Chinese medicine(CM)has become an important means in treating NAFLD in China.Intestinal microecology(IM)is an important part of the internal environment in the human body and is involved in the occurrence and development of NAFLD.In this paper,the authors systematically discuss the significance of IM in the pathogenesis of NAFLD and the current status of research on the CM treatment of NAFLD via IM regulation.In combination with our own research practice,we propose that IM is an important target for the treatment of NAFLD with CM and formulate plans for future research to target limitations existing in current studies.

Metabolic profiling of endogenous bile acids:a novel method to assess hepatoprotective effect of Tanreqing capsule on carbon-tetrachloride-induced liver injury in rats

Tanreqing(TRQ), a traditional Chinese medicine(TCM) formula, can alleviate liver injury and improve liver function. Its pharmacological mechanisms of actions are still unclear due to its complex components and multi-target natures. Metabolomic study is an effective approach to investigating drug pharmacological actions, new diagnostic markers, and potential mechanisms of actions. In the present study, a new strategy was used to evaluate the protective effect of TRQ capsule against carbon tetrachloride(CCl_4)-induced hepatotoxicity in rats, by analyzing metabolic profiling of endogenous bile acids(BAs) along with biochemical and histological analyses. BAs concentrations were determined by ultra-performance liquid chromatography coupled with quadrupole mass spectrometry(UPLC-MS). Principal component analysis and partial least squares discriminant analysis were then employed to analyze the UPLC-MS results and compare the hepatoprotective effect of TRQ capsule in different groups at the doses of 0.36, 1.44, and 2.88 g·kg^(-1) body weight, respectively. Moreover, our results suggested that taurocholic acid(TCA) and taurohyodesoxycholic acid(THDCA) were the most important biochemical markers, which were indicative of CCl_4-induced acute hepatic damage and hepatoprotective effect of TRQ capsule. Therefore, this new strategy would be an excellent alternative method for evaluating hepatoprotective effect and proposing potential mechanisms of action for other drugs as well.

Chinese Medicine Treatment of Refractory Ascites with Pleural Effusion Caused by Alcoholic Liver Disease: A Case Report

Alcohol is one of the leading causes of chronic liver disease and liver-related deaths worldwide.In particular,an overall high average level of alcohol consumption and heavy drinking binges appear to be critical factors that lead to disease and injury.Herein,we report a case who was diagnosed with refractory ascites and pleural ffusion caused by liver cirrhosis due to alcoholic liver disease.The patient was given diuretic therapy,antibiotics,reinfusion of concentrated ascites,and thoracic close drainage,but the result was disappointing.However,he responded well to Chinese medicine(CM)treatment.

Hepatic transcriptome delineates the therapeutic effects of Sanren Tang(三仁汤)on high-fat diet-induced non-alcoholic fatty liver disease

OBJECTIVE:To evaluate the effects of Sanren Tang(SRT,三仁汤)on a high-fat diet(HFD)-induced non-alcoholic fatty liver disease(NAFLD)in mice and to investigate the hepatic transcriptome regulated by SRT.METHODS:The primary SRT components were identified using ultra-high-performance liquid chromatography-high-resolution accurate mass spectrometry.The SRT-induced pharmacological effects on HFD-induced NAFLD were evaluated in mice for 16 weeks.Obeticholic acid was used as a control drug.Body weight,food intake,and homeostatic model assessment for insulin resistance(HOMA-IR)index were analysed.Hepatic histological changes were observed in haematoxylin and eosin-stained sections and quantified using the NAFLD activity score(NAS).Serum alanine aminotransferase(ALT)and hepatic triglyceride(TG)levels were measured.Lipids in hepatocytes were visualised by Oil red staining.RNA-sequencing was performed to determine the transcriptome profile of the liver tissue.The differentially expressed genes were validated using real-time polymerase chain reaction and Western blotting.RESULTS:Four principal compounds were identified in the SRT:adenosine,amygdalin,luteoloside,and magnolol.SRT ameliorated hepatic histology and lipid deposition in the NAFLD mice,and decreased HOMA-IR,NAS and ALT,and hepatic TG levels.Hepatic transcriptome analysis revealed 232 HFD-regulated genes that were reversed by SRT simultaneously.Retinol metabolism,cytokine-cytokine receptor interaction,and peroxisome proliferator-activated receptor(PPAR)γsignalling were the top three SRT-regulated pathways in NAFLD.CONCLUSIONS:SRT significantly ameliorated HFD-induced NAFLD,which was correlated with the regulation of genes enriched in the retinol metabolism,cytokine-cytokine receptor interaction,and PPARγsignalling pathways.

What are the challenges facing cancer therapy base on the dissipative structure theory

Cancer therapy is facing challenges.Since current theory could not have a breakthrough,we ought to find a new way to conquer cancer by exploring the emerging theory in cancer therapy.The complexity science,emerging in the 1980s,is the new stage of the system science development.It raises a series of transformation from humanities to natural science.Dissipative structure theory as a major science of complexity science,when applied to medical field,probable provide opportunity and inspiration referring to treatment.Dissipative structure theory,in brief,claims that a dissipative structure transform chaos to relatively stable non-equilibrium state through non-equilibrium phase transition.From the microscopic order,cancer composed by tumor cells is a dissipative structure,whose external environment is the human body.From the macroscopic order,the human body is a dissipative structure,whose external environment is the natural and social environment.The reason for tumor occurrence and progression is relatively non-equilibrium disrupted and negative entropy is not enough to correct high positive entropy.Survival with the tumor is to enhance negative entropy to resist highly positive entropy and then build a new relative non-equilibrium state.Innovative angle from the theory may enlighten us make the better use of dissipative structure theory in clinical cancer therapy.

Danhongqing formula alleviates cholestatic liver fibrosis by downregulating long non-coding RNA H19 derived from cholangiocytes and inhibiting hepatic stellate cell activation

Objective:This study explores the mechanism of action of Danhongqing formula(DHQ),a compoundbased Chinese medicine formula,in the treatment of cholestatic liver fibrosis.Methods:In vivo experiments were conducted using 8-week-old multidrug resistance protein 2 knockout(Mdr2-/-)mice as an animal model of cholestatic liver fibrosis.DHQ was administered orally for 8 weeks,and its impact on cholestatic liver fibrosis was evaluated by assessing liver function,liver histopathology,and the expression of liver fibrosis-related proteins.Real-time polymerase chain reaction,Western blot,immunohistochemistry and other methods were used to observe the effects of DHQ on long non-coding RNA H19(H19)and signal transducer and activator of transcription 3(STAT3)phosphorylation in the liver tissue of Mdr2-/-mice.In addition,cholangiocytes and hepatic stellate cells(HSCs)were cultured in vitro to measure the effects of bile acids on cholangiocyte injury and H19 expression.Cholangiocytes overexpressing H19 were constructed,and a conditioned medium containing H19 was collected to measure its effects on STAT3 protein expression and cell activation.The intervention effect of DHQ on these processes was also investigated.HSCs overexpressing H19 were constructed to measure the impact of H19 on cell activation and assess the intervention effect of DHQ.Results:DHQ alleviated liver injury,ductular reaction,and fibrosis in Mdr2-/-mice,and inhibited H19expression,STAT3 expression and STAT3 phosphorylation.This formula also reduced hydrophobic bile acid-induced cholangiocyte injury and the upregulation of H19,inhibited the activation of HSCs induced by cholangiocyte-derived conditioned medium,and decreased the expression of activation markers in HSCs.The overexpression of H19 in a human HSC line confirmed that H19 promoted STAT3 phosphorylation and HSC activation,and DHQ was able to successfully inhibit these effects.Conclusion:DHQ effectively alleviated spontaneous cholestatic liver fibrosis in Mdr2-/-mice by inhibiting H19 upregulation in cholangiocytes and preventing the inhibition of STAT3 phosphorylation in HSC,thereby suppressing cell activation.