Biological connotation of "phlegm-turbidity" theory in hepatocellular carcinoma microenvironment based on abnormal lipid metabolism / 临床肝胆病杂志

There is still a lack of effective strategies for the prevention and treatment of liver cancer, and a deep understanding of its pathogenesis may help to develop new treatment methods. Due to the abnormal changes of lipid metabolism in the development and progression of liver cancer, such process is closely associated with the "phlegm-turbidity" theory in traditional Chinese medicine (TCM). Starting from the changes of lipid metabolism in hepatocellular carcinoma microenvironment, this article discusses the association of the abnormal changes of lipid metabolism in tumor cells and immune cells with the "phlegm-turbidity" theory and the clinical efficacy of phlegm-eliminating therapies in clinical practice. Since the "phlegm-turbidity" theory in TCM plays an important role in the pathogenesis and pathological changes of liver cancer, the analysis of its theoretical connotation helps to clarify pathological mechanism, thereby providing a theoretical basis for the role of TCM in the prevention and treatment of liver cancer.

Distribution of traditional Chinese medicine syndrome types and elements in liver cirrhosis patients with dysplastic nodules: An analysis of 138 cases / 临床肝胆病杂志

Objective To investigate the distribution of traditional Chinese medicine (TCM) syndrome types and elements in liver cirrhosis patients with dysplastic nodules (DN), and to provide a basis for exploring the connotation and pattern of TCM syndrome types of DN in liver cirrhosis. Methods A total of 138 patients who attended The First Affiliated Hospital of Henan University of Chinese Medicine from March 2013 to January 2021 and were diagnosed with liver cirrhosis and DN were enrolled. General data such as age of onset and sex were collected, as well as the data on etiology, TCM syndrome types, and Child-Pugh class for liver function, and the distribution characteristics of TCM syndrome types and elements were summarized. The chi-square test or the Fisher's exact test was used for comparison of categorical data between groups. Results The liver and the spleen were the main syndrome elements of disease location in liver cirrhosis patients with DN, accounting for 97.83% and 94.93%, respectively, followed by the kidney (23.91%); Qi deficiency and Qi stagnation were the main syndrome elements reflecting the nature of disease, accounting for 73.91% and 58.70%, respectively, followed by dampness (34.78%). The main TCM syndrome types included stagnation of liver Qi and spleen deficiency, damp-heat internal excess syndrome, blood stasis and toxin accumulation syndrome, and water-dampness retention syndrome, among which stagnation of liver Qi and spleen deficiency was more common and accounted for 58.70% ( P 0.05). There was a significant difference in Child-Pugh class between the liver cirrhosis DN patients with different TCM syndrome types ( χ 2 =34.320, P < 0.05), and Child-Pugh class A was more common in the patients with stagnation of liver Qi and spleen deficiency (59.8%), while Child-Pugh class C was more common in the patients with damp-heat internal excess syndrome (39.1%). Conclusion This article summarizes the distribution characteristics of common TCM syndrome types and elements of DN in liver cirrhosis, which provides a reference for the syndrome differentiation-based TCM treatment of DN in liver cirrhosis.

A case analysis of acute-on-chronic liver failure caused by abiraterone / 中国药房

Abiraterone is commonly used as a targeted drug for the treatment of prostate cancer， which commonly causes adverse drug reactions （ADR）， including abnormal liver function， fatigue， nausea and edema， etc. This study reports a 78-year-old man with a history of hepatitis B and liver cirrhosis after prostate cancer resection who was admitted to the First Affiliated Hospital of Henan University of Chinese Medicine. The patient received abiraterone treatment 1 month before admission and developed gastrointestinal symptoms 3 weeks after the treatment and worsened at 4th week with yellowing of the skin， sclera and urine. Unfortunately， the patient died after 5 weeks of abiraterone treatment （1 week after admission）. Based on test and examination results， the patient was diagnosed with acute-on-chronic liver failure （ACLF）. This paper analyzes the patient’s medical history and the relevant treatment in detail. It is evaluated that ACLF and abiraterone are “probably” related based on Naranjo ADR Probability Scale， suggesting abiraterone may induce severe ADR of liver failure in patients with chronic liver diseases such as cirrhosis. These patients should be monitored dynamically for changes in liver function and treated prophylactically with liver-protective drugs if necessary.

Amygdalin Ameliorates Liver Fibrosis through Inhibiting Activation of TGF-β/Smad Signaling / 中国结合医学杂志

OBJECTIVE@#To observe the effect of amygdalin on liver fibrosis in a liver fibrosis mouse model, and the underlying mechanisms were partly dissected in vivo and in vitro.@*METHODS@#Thirty-two male mice were randomly divided into 4 groups, including control, model, low- and high-dose amygdalin-treated groups, 8 mice in each group. Except the control group, mice in the other groups were injected intraperitoneally with 10% carbon tetrachloride (CCl4)-olive oil solution 3 times a week for 6 weeks to induce liver fibrosis. At the first 3 weeks, amygdalin (1.35 and 2.7 mg/kg body weight) were administered by gavage once a day. Mice in the control group received equal quantities of subcutaneous olive oil and intragastric water from the fourth week. At the end of 6 weeks, liver tissue samples were harvested to detect the content of hydroxyproline (Hyp). Hematoxylin and eosin and Sirius red staining were used to observe the inflammation and fibrosis of liver tissue. The expressions of collagen I (Col-I), alpha-smooth muscle actin (α-SMA), CD31 and transforming growth factor β (TGF-β)/Smad signaling pathway were observed by immunohistochemistry, quantitative real-time polymerase chain reaction and Western blot, respectively. The activation models of hepatic stellate cells, JS-1 and LX-2 cells induced by TGF-β1 were used in vitro with or without different concentrations of amygdalin (0.1, 1, 10 µmol/L). LSECs. The effect of different concentrations of amygdalin on the expressions of liver sinusoidal endothelial cells (LSECs) dedifferentiation markers CD31 and CD44 were observed.@*RESULTS@#High-dose of amygdalin significantly reduced the Hyp content and percentage of collagen positive area, and decreased the mRNA and protein expressions of Col-I, α-SMA, CD31 and p-Smad2/3 in liver tissues of mice compared to the model group (P<0.01). Amygdalin down-regulated the expressions of Col-I and α-SMA in JS-1 and LX-2 cells, and TGFβ R1, TGFβ R2 and p-Smad2/3 in LX-2 cells compared to the model group (P<0.05 or P<0.01). Moreover, 1 and 10 µmol/L amygdalin inhibited the mRNA and protein expressions of CD31 in LSECs and increased CD44 expression compared to the model group (P<0.05 or P<0.01).@*CONCLUSIONS@#Amygdalin can dramatically alleviate liver fibrosis induced by CCl4 in mice and inhibit TGF-β/Smad signaling pathway, consequently suppressing HSCs activation and LSECs dedifferentiation to improve angiogenesis.

Pathological Mechanism of HSOS in Rats Induced by Different Doses of Monocrotaline / 中国实验方剂学杂志

ObjectiveTo observe the pathological changes of hepatic sinusoidal obstruction syndrome （HSOS） induced by different doses of monocrotaline （MCT） in rats， investigate the dose and duration of modeling， and elucidate the mechanism. MethodA total of 72 male SD rats were randomized into normal group （n=12）， and low-， medium-， and high-dose MCT groups （n=20 per group， 80，120，160 mg·kg-1， respecctively）. In the model groups， different doses of MCT were intragastrically administered to induce the HSOS in rats. After 48 h and 120 h separately， rats in each group were sacrificed and sampling was performed. The survival rate of rats in each group was calculated， and the body weight， liver weight， and and serum liver function indexes of the rats were examined. The histopathological changes of the liver were observed based on scanning electron microscopy， hematoxylin and eosin （HE） staining， and Sirius red （SR） staining. Glutathione S-transferase （GST） activity， total superoxide dismutase （T-SOD） activity， and malondialdehyde （MDA） content of liver tissue homogenate were measured with microplate method. The expression of liver tissue-related indexes was detected by real-time polymerase chain reaction （PCR）， Western blot， and immunohistochemistry. ResultThe activity of serum alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in MCT groups rose with the increase in MCT dose （P<0.05， P<0.01） compared with that in the normal group. With the extension of modeling time， the activity of serum ALT and AST in the low-dose group decreased （P<0.01）， while the activity of them in the medium-dose and high-dose groups increased （P<0.01）. HE staining showed that hepatocyte necrosis， inflammatory cell infiltration， and erythrocyte accumulation in MCT groups. Electron microscopy demonstrated that fenestrae of liver sinusoidal endothelial cells widened and the sieve plates disappeared. Morever， the injury was worsened with the increase in MCT dose. In addition， the expression of CD44 in MCT groups was significantly reduced compared with that in the normal group （P<0.05， P<0.01）. SR staining showed that no positive staining was found in model groups after 48 h， while collagen deposition in portal areas and liver sinusoids could be seen in model groups after 120 h. MCT groups showed increase in MDA content and GST activity and decrease in T-SOD activity compared with the normal group， particularly the medium-dose and high-dose groups （P<0.01）， and the changes were dose-dependent after 120 h （P<0.01）. The protein expression of CD68 （pro-inflammatory macrophage marker） was raised with the increase in dosage， which was consistent with the results of immunohistochemistry （P<0.01）， while CD163 （anti-inflammatory macrophage marker） protein and mRNA expression was significantly decreased with the increase in dosage （P<0.01）. Western blot results showed that the expression of phosphorylated nuclear factor-κB/nuclear factor-κB （p-NF-κB/NF-κB） and phosphorylated protein kinase B/protein kinase B （p-Akt/t-Akt） was significantly increased in medium-dose and high-dose MCT groups （P<0.05，P<0.01）. The protein expression of α-smooth muscle actin （α-SMA） in liver tissues in MCT groups was significantly increased over time and with the increase in dose， and the mRNA expression of α-SMA， collagen type I α1 （Col1a1）， and collagen type Ⅳ α1 （Col4a1） showed the same trend （P<0.05， P<0.01）. The results of TUNEL staining showed that apoptotic cells were increased with the rise of MCT dose， while B-cell lymphoma-2（Bcl-2） /Bcl-2 associated X protein （Bax） was remarkably decreased （P<0.01）. ConclusionHSOS in rats induced by intragastric administration of different doses of MCT was aggravated with the increase of dosage. In the low-dose （80 mg·kg-1） MCT group， the liver healed spontaneously over time. However， liver damage caused by MCT of 120 mg·kg-1 and 160 mg·kg-1 aggravated over time， and even fibrosis and death occurred. The pathological mechanism of MCT-induced HSOS in rats may be that MCT triggered intense oxidative stress in liver tissue， thus activated pro-inflammatory macrophages to secrete large amounts of inflammatory factors， and further activated the NF-κB/Akt signalling pathway， leading to severe cell damage and death.

Medication rule of TCM patent compound prescriptions for the treatment of primary liver cancer based on data mining / 国际中医中药杂志

Objective:To analyze the medication rule of TCM compound prescription in the treatment of Primary Liver Cancer (PLC) in the National Patent Database by the method of data mining.Methods:By searching for the compound prescriptions of TCM which were included in the National Patent Retrieval Platform for the treatment of PLC, and analyzing the frequency, association and cluster of those included compound prescriptions through the inheritance and assistance platform of TCM.Results:A total of 350 patent prescriptions for the treatment of PLC were included, involving 425 Chinese medicines; the top 10 frequently appeared herbal medicines were Hedyotis diffusa, Radix Astragali, Scutellaria barbata, Atractylodes macrocephala, Glycyrrhiza uralensis, Ginseng, Radix Bupleuri, Rhizoma Curcumae, Salvia miltiorrhiza and Angelica sinensis; according to the function of those medicines, the top three frequently appeared herbal medicines were those with deficiency-tonifying heat-clearing and detoxifying, promoting blood circulation and removing blood stasis functions; the property of the medicine were mainly cold, warm and mild; the taste of the medicine were mainly bitter, sweet and acrid; the meridians of the medicines mainly belong to liver, spleen and lung. It was found that there were 12 association rules, and the medicine pair that were closely correlated was " Hedyotis diffusa- Scutellaria barbata", and the most frequently appeared medicines could be grouped into 8 groups. Conclusions:The TCM patent compound prescriptions mainly play the role of replenishing qi and invigorating the spleen, supplemented by the products of clearing heat and detoxification, activating blood circulation and removing blood stasis, which is consistent with the basic treatment principle of PLC "invigorate the upright and dispel the evil, attack the evil and tonify the body". This study explores the basic medication rules of TCM patent compound prescription in the treatment of PLC, in order to provide guidance for clinical practice.

Role of GDC-0449 in a rat model of liver fibrosis induced by carbon tetrachloride combined with 2-acetylaminofluorene / 临床肝胆病杂志

Objective To investigate the intervention effect of GDC-0449, a hedgehog signaling pathway inhibitor, on rats with liver fibrosis induced by carbon tetrachloride (CCl 4 ) combined with 2-acetylaminofluorene (2-AAF). Methods A total of 18 female Fisher344 rats were randomly divided into normal group, CCl 4 /2-AAF group, and GDC-0449 group, with 6 rats in each group. The rats in the CCl 4 /2-AAF group and the GDC-0449 group were given subcutaneously injected 30% CCl 4 -olive oil solution at a dose of 2 mL/kg twice a week for 6 weeks to induce liver fibrosis; since week 7, in addition to the injection of CCl 4 -olive oil solution, the rats in these two groups were given 2-AAF (100 mg/kg/d) by gavage, and the rats in the GDC-0449 group were given GDC-0449 (25 mg/kg/d) by gavage, while those in the normal group were given an equal volume of olive oil solution by injection and normal saline by gavage. All rats were sacrificed at the end of week 9, and related samples were collected. HE staining and sirius red (SR) staining were used to observe the changes in liver histopathology and collagen deposition, and the semi-quantitative analysis of SR-positive area and Ishak score were used to evaluate fibrosis degree; the alkaline hydrolysis method was used to measure the level of hydroxyproline (Hyp) in liver tissue; immunohistochemistry, Western blot, and qRT-PCR were used to measure the expression of α-smooth muscle actin (α-SMA), type Ⅰ collagen (Col-Ⅰ), type Ⅳ collagen (Col-Ⅳ), cytokeratin 19 (CK19), cytokeratin 7 (CK7), the epithelial cell adhesion molecule Epcam, and the hedgehog signaling pathway in liver tissue; double immunofluorescence staining was used to observe the colocalization of CK19 and the oval cell marker OV6. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results Compared with the normal group, the CCl 4 /2-AAF group had marked inflammatory cell aggregation and collagen deposition in liver tissue, with the formation of a pseudolobular structure, as well as significant increases in Hyp level and collagen positive area ratio in liver tissue ( P < 0.05), Ishak score ( P < 0.05), and the expression of α-SMA, Col-Ⅰ, Col-Ⅳ, Epcam, CK19, CK7, the transmembrane transporter Smoothened (Smo), Hedgehog ligand Desert Hedgehog (Dhh), the Indian Hedgehog membrane-binding receptor Patched (Ptch2), and glioma-related oncogenes Gli1, Gli2, and Gli3 (all P < 0.05); double immunofluorescence staining showed that CK19-positive cells also expressed OV6 in the liver tissue of rats in the CCl 4 /2-AAF group, with a significant increase compared with the normal group. Compared with the CCl 4 /2-AAF group, the GDC-0449 group had significant reductions in inflammatory cell aggregation and collagen deposition in liver tissue, Hyp level and collagen positive area ratio in liver tissue ( P < 0.05), Ishak score ( P < 0.05), and the expression of α-SMA, Epcam, CK19, CK7, Smo, Ptch2, Gli1, Gli2, and Gli3 (all P < 0.05); double immunofluorescence staining showed a significant reduction in the number of cells with co-expression of OV6 and CK19 in liver tissue. Conclusion The Hedgehog signaling pathway inhibitor GDC-0449 can significantly inhibit the progression of liver fibrosis induced by CCl 4 /2-AAF in rats, possibly by inhibiting hepatic stellate cell activation, collagen deposition, activation and proliferation of hepatic progenitor cells, and differentiation of hepatic progenitor cells into biliary epithelial cells.

Mechanism of ferroptosis in the formation of nonalcoholic fatty liver disease/nonalcoholic steatohepatitis / 临床肝胆病杂志

Ferroptosis is a type of iron-dependent cell death driven by lipid peroxidation, and its mechanism is associated with iron homeostasis imbalance, lipid peroxidation, and slC7A11-GSH-GPX4 antioxidant system. Ferroptosis plays a key role in the development and progression of nonalcoholic fatty liver disease (NAFLD)/nonalcoholic steatohepatitis (NASH), and inhibition of ferroptosis can almost completely inhibit the development of NASH. This article reviews the research advances in the mechanism of ferroptosis and its role in NAFLD/NASH and proposes the research strategies and technical means for ferroptosis, so as to provide a reference for research on the mechanism of NAFLD/NASH.

Vascular endothelial growth factor expression and angiogenesis in human ovarian tissue after different cryopreservations / 中国组织工程研究

BACKGROUND: The cryopreservation of human ovarian tissue has become an attractive method to preserve female fertility. Human ovarian tissue experiences neovasculadzation after transplantation to recover blood supply, cryopreservation and resuscitation technique is a key for the neovascularization of human ovarian tissue following transplantation. OBJECTIVE: To investigate vascular endothelial growth factor (VEGF) expression and microvessel density in human ovadan tissue following novel needle immersed vitrification (NIV) and slow-freezing, to explore the influence of two cryopreservation methods play in the neovascularization of human ovarian tissue after transplantation. METHODS: Eight normal human ovarian tissues from patients with carcinoma of endometrium were cut into 12 fragments in the size of 1.5 mm × 1.5 mm × 1.0 mm, then randomly assigned to 3 groups: fresh control group, NIV group and slow-freezing group. In the NIV group, pieces of ovarian tissue stdps were dehydrated in an equilibration solution consisting of 7.5% ethylene glycol and 7.5% dimethyl sulfoxide in TCM-199 supplement with 20% fetal bovine serum and a vitrification solution consisting of 15% ethylene glycol, 15% dimethyl sulfoxide and 0.5 mol/Lsucrose, then were plunged in liquid nitrogen directly and sealed in liquid nitrogen-filled cryovials. For thawing, the needles holding ovadan tissues were serially transferred into 1.0, 0.5, 0.25 mol/L sucrose solution and TCM-199 supplemented with 20% fetal bovine serum. In the slow-freezing group, pieces of human ovadan cortex fragments were placed in a 1.8-mL cryovial containing 1 mL of TCM-199 medium supplemented with 0.1 mol/L sucrose, 20% fetal bovine serum and 1.5 mol/L dimethyl sulfoxide, the cryovials were placed in the programmable freezer and cryopreserved by preset slow-cooling protocol. For thawing, the ovarian tissue stdps were washed in a stepwise manner: 1.0 mol/L dimethyl sulfoxida + 0.1 mol/L sucrose, 0.5 mol/L dimethyl sulfoxide + 0.1 mol/L sucrose, 0.25 mol/L dimethyl sulfoxJde + 0.1 mol/L sucrose and 0.1 mol/L sucrose. The frozen-thawed and fresh controlled human ovarian tissues were cultured in vitro. The expression of VEGF and CD34, as well as microvessel density, was analyzed by immunohistochemistry. RESULTS AND CONCLUSION: There was patchy and mild expression of VEGF in the stromal cells of all the three groups before and after culture. The expression of VEGF increased and reached peak value after culture for 2 days, began to decrease after culture for 4 days and further attenuated after culture for 6 days in all the three groups. Compared with slow-freezing group, the expression of VEGF in NIV group was closer to that in fresh control group. Microvessel density of all the three groups increased and reached peak value after culture for 2 days, and the microvessel density of fresh control group and NIV group was significantly higher than that of slow-freezing group (P < 0.05). The microvessel density of slow-freezing group after culture for 4 days and that of all the three groups after culture for 6 days significantly decreased compared with after culture for 2 days (P <0.05). NIV is superior to slow-freezing to preserve stromal cells and extracellular matrix of human ovarian tissue, and plays less influence in VEGF expression and angiogenesis in human ovarian tissue.