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ObjectiveTo explore the mechanism and pathway of Gandou Fumu decoction (GDFMD) in the development of liver fibrosis in Wilson's disease (WD). MethodFirst, 30 TX-j mice were randomly divided into the model group, high-dose, medium-dose, and low-dose GDFMD groups, and penicillamine group, with six mice in each group, and another six wild-type mice were used as the normal group. The high-dose, medium-dose, and low-dose GDFMD groups were intragastrically administered drugs of 13.92, 6.96, 3.48 g·kg-1. In the penicillamine group, 0.1 g·kg-1 of penicillamine was given by intragastric administration. The model group and the normal group were given equal volume of normal saline, once a day, for four consecutive weeks. Samples were collected four weeks after gavage, and enzyme-linked immunosorbent assay (ELISA) was used to detect type Ⅲ procollagen peptide (PCⅢ), collagen type Ⅳ (Col Ⅳ), hyaluronic acid (HA), and laminin (LN). Hematoxylin-eosin (HE), Masson, and picric acid-Sirus red collagen (Sirus Red) staining were used to observe the histopathological changes of liver fibrosis. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), immunohistochemistry, and Western blot were used to observe the expressions of α-smooth muscle actin (α-SMA) and collagen type Ⅰ (Col Ⅰ), which were related to the activation of hepatic stellate cells (HSCs). The expression of miR-29b-3p was observed by Real-time PCR. The expression of Unc-51-like kinase 1 (ULK1) and its downstream-related factors were observed by Western blot. The downstream genes of miR-29b-3p were verified by the dual luciferase reporter gene detection method. ResultCompared with the normal group, the four items of liver fibrosis (PCⅢ, Col Ⅳ, HA, and LN) in the model group were significantly abnormal (P<0.01), and the pathology was significantly abnormal. The expression of HSC activation-related indicators including α-SMA and Col Ⅰ, as well as α-SMA mRNA and Col Ⅰ mRNA was up-regulated (P<0.05, P<0.01), and miR-29b-3p expression was down-regulated (P<0.01). ULK1, p-ULK1, autophagy-related gene 13 (Atg13), p-Atg13, Beclin-1, FAK family kinase-interacting protein of 200 kDa (FIP200), activating molecule in BECN1-regulated autophagy protein 1 (AMBKA1), and microtubule-associated protein 1 light chain 3Ⅱ/Ⅰ(LC3Ⅱ/Ⅰ) were up-regulated (P<0.05, P<0.01). p62 protein expression was down-regulated (P<0.01). Compared with the model group, the four items of liver fibrosis in the high-dose, medium-dose, and low-dose GDFMD groups and the penicillamine group were significantly improve (P<0.01), and the pathological conditions were improved. The expression of HSC activation-related indicators including α-SMA and Col Ⅰ, as well as α-SMA mRNA and Col Ⅰ mRNA was down-regulated (P<0.05, P<0.01), and the expression of miR-29b-3p was up-regulated (P<0.01). ULK1, p-ULK1, Atg13, p-Atg13, Beclin-1, FIP200, AMBKA1, and LC3Ⅱ/Ⅰ were down-regulated (P<0.05, P<0.01), and p62 protein expression was up-regulated (P<0.01). The prediction software predicted that there was a binding site between miR-29b-3p and ULK1. The dual-luciferase reporter gene detection method indicated that the luciferase activity of the ULK1-WT plasmid-transfected cell group was reduced when miR-29b-3p mimics were co-cultured (P<0.01). ConclusionGDFMD can regulate ULK1-mediated autophagy by up-regulating miR-29b-3p and further exert its anti-hepatic fibrosis effect in Wilson's disease.
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The syndrome differentiation of Yin and Yang has the function of controlling the other six principles in the eight principles syndrome differentiation,which is a higher level or general induction of the disease. In the clinical process of traditional Chinese medicine,syndrome differentiation of Yin and Yang runs through the whole process of disease diagnosis and treatment. For Parkinson's disease,syndrome differentiation of Yin and Yang is particularly important. Different symptoms,the transformation of pathogenesis during the development of the disease and the treatment of traditional Chinese and western medicine all reflect the characteristics of Yin and Yang opposition restriction,mutual root and mutual use,and the transformation of ebb and flow. This article discusses the background,application and value of Yin-Yang syndrome differentiation from three aspects:the origin and application of yin-yang syndrome differentiation,the basis of Parkinson's disease syndrome differentiation,and the status and role of Yin-Yang syndrome differentiation in Parkinson's disease. It is of great significance to guide the diagnosis and treatment of Parkinson's disease with "Yin-Yang as the key point".
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Wilson's disease (WD) is a copper metabolism disorder caused by mutations in the ATP7B gene, with diverse phenotypes and complex pathogenesis. It is one of the few rare diseases that can achieve good clinical efficacy through standardized treatment. Since there are few systematic reviews of this disease, we summarize the pathogenesis and treatment methods of WD from traditional Chinese and western medicine by reviewing the literature related to WD. In western medicine, ATP7B gene mutation is considered as the root cause of WD, which affects copper transport and causes copper metabolism disorders. The excessive copper deposited in the body will result in oxidative stress, defects in mitochondrial function, and cell death. Western medicine treatment of WD relies mainly on drugs, and copper antagonists are the first choice in clinical practice, which are often combined with hepatoprotective and antioxidant therapy. Surgery is a common therapy for the patients with end-stage WD, and gene therapy provides an option for WD patients. According to the traditional Chinese medicine (TCM) theory, WD is rooted in constitutional deficiency and copper accumulation and triggered by dampness-heat accumulation or phlegm combined with stasis. The patient syndrome varies in different stages of the disease, and thus the treatment should be based on syndrome differentiation. The TCM treatment method of nourishing the liver and kidneys and warming the spleen and kidneys can address the root cause. The methods of clearing heat and drying dampness, resolving phlegm and dispelling stasis, and soothing liver and regulating qi movement can be adopted to treat symptoms. On the basis of syndrome differentiation, special prescriptions for the treatment of WD have been formulated, such as Gandou decoction, Gandouling, and Gandou Fumu decoction, which have been widely used in clinical practice. TCM and western medicine have their own advantages and shortcomings. The integrated Chinese and western medicine complementing with each other demonstrates great therapeutic potential. This paper summarizes the pathogenesis and treatment of WD with integrated Chinese and western medicine, aiming to provide a reference for the clinical diagnosis and treatment of this disease.
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ObjectiveTo investigate the protective effect and underlying mechanism of Gandou Fumu decoction (GDFMT) on renal fibrosis in a mouse model of Wilson's disease. MethodSixty adult male toxic milk (TX) mice were randomly divided into a model group, high-, medium-, and low-dose GDFMT groups, and a positive control (penicillamine) group, and another 12 wild-type mice were assigned to the normal group. The high-, medium-, and low-dose GDFMT groups were administered GDFMT at 13.92, 6.96, 3.48 g·kg-1, respectively, and the positive control group received penicillamine at 0.1 g·kg-1, while the model and normal groups were given an equal volume of 0.9% saline solution by gavage once a day for 4 consecutive weeks. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of blood urea nitrogen (BUN), creatinine (CRE), type Ⅲ procollagen (PC-Ⅲ), and type Ⅳ collagen (C-Ⅳ) in the serum. Histological changes in the mouse kidneys were examined by hematoxylin-eosin (HE) and Masson's trichrome staining. Immunofluorescence was used to assess the protein expression of leptin, Janus kinase 2 (JAK2), and signal transducer and activator of transcription (STAT) in renal cells. Real-time polymerase chain reaction (Real-time PCR) was performed to analyze the mRNA expression levels of leptin, leptin receptor(OB-R), JAK2, and STAT. Western blot was used to detect the expression of transforming growth factor-β1 (TGF-β1) and monocyte chemoattractant protein-1 (MCP-1). ResultCompared with the normal group, the model mice exhibited a significant increase in BUN, CRE, PC-Ⅲ, and C-Ⅳ levels (P<0.01). Compared with the model group, the high- and medium-dose GDFMT groups and the penicillamine groups showed significant decreases in these parameters (P<0.05, P<0.01), with the high-dose GDFMT group demonstrating the most significant reduction (P<0.01). The histological examination of renal tissue revealed fibrosis in the model group, while the fibrotic damage was mitigated to varying degrees after drug intervention, with improvement in fibrosis. Immunofluorescence results showed that leptin, JAK2, and STAT3 protein expression levels were significantly upregulated in the renal fibrosis of the model group. After GDFMT intervention, the fluorescence intensity decreased, with the high-dose GDFMT group showing the lowest intensity. Real-time PCR results demonstrated that leptin, OB-R, JAK2, and STAT3 mRNA expression levels were significantly elevated in the model group compared with those in the normal group, while the high- and medium-dose GDFMT groups and the penicillamine group showed significant reductions in their expression levels (P<0.05, P<0.01). Western blot analysis revealed that TGF-β1 and MCP-1 expression levels were significantly increased in the model group (P<0.01), and the high- and medium-dose GDFMT groups exhibited significant reductions in their expression levels (P<0.01). ConclusionGDFMT can alleviate renal fibrosis damage in TX mice, and its mechanism of action may be related to the regulation of leptin and the JAK/STAT signaling pathway.
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Wilson's disease (WD) is a rare autosomal recessive disorder with a complex pathogenesis involving multiple systems, multiple visceral organs, and the complex copper homeostasis regulation system within the body. The liver is the most common organ for copper deposition, and liver injury is the earliest and most common manifestation of WD; therefore, it is important to find an ideal animal model for WD research. By summarizing the animal models of WD commonly used in the world, this article systematically summarizes the background, liver and nervous manifestations, and application of different models and compares the characteristics of different animal models, so as to provide a reference for the application of various animal models of WD.
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Liver fibrosis is the initial stage of the development of various chronic liver diseases into liver cirrhosis and is a reversible process. As a subset of extracellular vesicles that can carry active substances such as proteins, lipids, and RNA, exosomes are involved in intercellular signal communication and have attracted more and more attention in recent years. Studies have shown that non-coding RNAs in exosomes play an important role in the development and progression of liver fibrosis. This article discusses the mechanism of action of exosome long non- coding RNAs (including MALAT1, H19, GAS5, MEG3, PVT1, and P21), exosome short non-coding RNAs (including micro-RNA, small nucleolus RNA, PIWI-interacting RNA, and small interference RNA), and exosome circular RNA in the development and progression of liver fibrosis, and it is concluded that exosomes from different sources (such as hepatocytes and cholangiocytes) carrying non-coding RNAs mainly affect the activation, proliferation, migration, and transformation of hepatic stellate cells. In-depth studies of exosome non-coding RNAs in the future are expected to find potential new targets for the treatment of liver fibrosis.
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Liver fibrosis is the initial stage of the development of various chronic liver diseases into liver cirrhosis and is a reversible process. As a subset of extracellular vesicles that can carry active substances such as proteins, lipids, and RNA, exosomes are involved in intercellular signal communication and have attracted more and more attention in recent years. Studies have shown that non-coding RNAs in exosomes play an important role in the development and progression of liver fibrosis. This article discusses the mechanism of action of exosome long non- coding RNAs (including MALAT1, H19, GAS5, MEG3, PVT1, and P21), exosome short non-coding RNAs (including micro-RNA, small nucleolus RNA, PIWI-interacting RNA, and small interference RNA), and exosome circular RNA in the development and progression of liver fibrosis, and it is concluded that exosomes from different sources (such as hepatocytes and cholangiocytes) carrying non-coding RNAs mainly affect the activation, proliferation, migration, and transformation of hepatic stellate cells. In-depth studies of exosome non-coding RNAs in the future are expected to find potential new targets for the treatment of liver fibrosis.
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Objective To observe the influence of chitosan on the skin and soft tissue expansion.Methods Twenty-five patients were selected,who were suitable to be embedded soft tissue expanders in the face,a 100-milliliter expander was implanted in one side of the face,and other side was used as control.A 100-milliliter expander was implanted in each group,and a slender silicon duct was embedded between the expander and subcutaneous tissue in the experimental group.About five to seven days after the operation,the negative drainage tube was removed,and then two-milliliter medical chitosan injected with the silicon duct in the experimental group,but not in the control group.Two groups were injected with normal saline in the second day.The center of expanded skin was pressed and skin elasticity and relaxation were compared between the two groups during the injection interval.The time of injection interval,the quantity of normal saline inside the expanders after two weeks and three weeks and the total time of expansion to 100 milliliters were recorded.After injection was completed in the two groups and maintained for two weeks.In the stage Ⅱ operation,the expanders were taken out,1 cm × 1 cm fibropeplos was removed from the center of the expanded skin flap from the two groups,and pathological section was prepared to measure the thickness of fibropeplos,average gray scale of collagen and the quantity of blood capillaries.The fibroblasts,collagen fiber and capillaries were observed and compared under light microscope.A matched-pairs t analysis was used to analyze the data.Results Compared with the control group,the quantity of normal saline inside the expanders in the experimental group was increased at the same time; the water injection period was shorten and tissue expansion was significantly accelerated.The number of fibroblasts in the fibropeplos decreased with the influence of chitosan.The fibroblasts were restrained to mature period and collegan decreased.The fibropeplos became thinner but the capillaries were not affected.Conclusions Chitoson can inhibit fibroblast proliferation and collagen production,and the effect of accelerating tissue expansion is significant and deserves to be recommended.