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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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OBJECTIVE To construct a risk prediction model for bloodstream infection （BSI） induced by carbapenem-resistant Klebsiella pneumoniae （CRKP）. METHODS Retrospective analysis was conducted for clinical data from 253 patients with BSI induced by K. pneumoniae in the First Hospital of Qinhuangdao from January 2019 to June 2022. Patients admitted from January 2019 to December 2021 were selected as the model group （n＝223）， and patients admitted from January 2022 to June 2022 were selected as the validation group （n＝30）. The model group was divided into the CRKP subgroup （n＝56） and the carbapenem- sensitive K. pneumoniae （CSKP） subgroup （n＝167） based on whether CRKP was detected or not. The univariate and multivariate Logistic analyses were performed on basic information such as gender， age and comorbid underlying diseases in two subgroups of patients； independent risk factors were screened for CRKP-induced BSI， and a risk prediction model was constructed. The established model was verified with patients in the validation group as the target. RESULTS Admissioning to intensive care unit （ICU）， use of immunosuppressants， empirical use of carbapenems and empirical use of antibiotics against Gram-positive coccus were independent risk factors of CRKP-induced BSI （ORs were 3.749， 3.074， 2.909， 9.419， 95%CIs were 1.639-8.572， 1.292- 7.312， 1.180-7.717， 2.877-30.840， P＜0.05）. Based on this， a risk prediction model was established with a P value of 0.365. The AUC of the receiver operating characteristic （ROC） curve of the model was 0.848 [95%CI （0.779， 0.916）， P＜0.001]， and the critical score was 6.5. In the validation group， the overall accuracy of the prediction under the model was 86.67%， and the AUC of ROC curve was 0.926 [95%CI （0.809， 1.000]， P＜0.001]. CONCLUSIONS Admission to ICU， use of immunosuppressants， empirical use of carbapenems and empirical use of antibiotics against Gram-positive coccus are independent risk factors of CRKP- induced BSI. The CRKP-induced BSI risk prediction model based on the above factors has good prediction accuracy.
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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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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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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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Objective:To explore the risk factors of mortality in patients with Klebsiella pneumoniae bloodstream infection, and to construct a predictive model. Methods:The clinical data of 234 patients with Klebsiella pneumoniae bloodstream infection admitted in the First Hospital of Qinhuangdao from January 2020 to December 2022 were retrospectively analyzed, including 202 cases admitted during January 2020 to June 2022 (model set), and 32 cases admitted during July to December 2022 (validation set). There were 64 cases died (fatal group) and 138 cases survived (survival group) within 28 d after admission in model set. Multivariate Logistic regression was used to analyze the risk factors of death in patients with Klebsiella pneumoniae bloodstream infection and a mortality prediction model was constructed. The constructed model was applied in validation set, and the consistency between predicted mortality and real mortality was analyzed. Results:Multivariate Logistic regression analysis showed that male sex ( OR=2.598, 95% CI 1.179-5.725, P=0.018), age≥65 years ( OR=4.420, 95% CI 2.029-9.627, P<0.001), admitted to intensive care unit (ICU) ( OR=10.299, 95% CI 4.752-22.321, P<0.001), and the empirical use of quinolones antibiotics ( OR=4.288, 95% CI 1.127-16.317, P=0.033) were independent risk factors for 28-day mortality in Klebsiella pneumoniae bloodstream infection patients. The regression equation for predicting the risk of death was -3.469+ male × 0.955+ age ≥ 65 years × 1.486+ admitted to ICU × 2.332+ empirical use of quinolone antibiotics × 1.456. The area under the ROC curve (AUC) for predicting death in the model set was 0.831, with sensitivity and specificity of 71.9% and 80.4%, respectively. The AUC for predicting death in the validation set was 0.881, with sensitivity and specificity of 91.7% and 75.0%, respectively. Conclusion:The constructed mortality prediction model in the study has good application value for the prognosis of patients with Klebsiella pneumoniae bloodstream infection.
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Objective:To observe the effects of high copper diet on neurobehavioral functions and synaptic associated protein expression in hippocampus of rats.Methods:Thirty male SD rats were randomly divided into control group and high copper diet group with 15 rats in each group according to the random number table method. The rats in control group were fed with ordinary diet and ordinary water, while the rats in high-copper diet group were fed with high-copper diet containing 1 g/kg copper sulfate and 0.185% copper sulfate deionized water for 12 weeks. The content of copper in serum and hippocampus of rats were detected by inductively coupled plasma-atomic emission spectrometry(ICP-AES) and ICP-mass spectrometry(ICP-MS). The neurobehavioral indicators were detected by stereotypic behavior test, open field test and Morris water maze test. The expression levels of microtubule associated protein 2(MAP2) and growth associated protein 43 (GAP43) in hippocampus were detected by Western blot.SPSS 22.0 software was used for statistical analysis, and two independent sample t-test was used for comparison between the two groups. Results:Compared with the control group, the content of serum copper((1.67±0.69)mg/L, (1.98±0.24)mg/L, t=17.53, P<0.05) and hippocampal free copper((3.52±1.24)mg/g, (4.78±0.57)mg/g, t=10.34, P<0.05) in the high copper diet group increased significantly, and the stereotypic behavior score increased significantly ((0.29±0.08), (2.97±0.72), t=14.33, P<0.01), the number of space crossing in the open field experiment ((153.40±24.73)points, (92.46±19.46)points, t=7.50, P<0.01) and the times of standing((19.34±1.98)times, (10.57±2.71)times, t=10.12, P<0.01) were significantly decreased. The average latency in Morris water maze navigation test was significantly prolonged ((3.14±1.67)s, (8.29±2.26)s, t=7.10, P<0.01), the number of crossing the original platform position in the space exploration test decreased significantly ((7.89±2.48)times, (2.98±1.73) times, t=3.23, P<0.01). Compared with control group, protein levels of GAP43((1.03±0.05), (0.48±0.02), t=39.56, P<0.05)and MAP2((0.93±0.05), (0.30±0.08), t=25.86, P<0.05) of high copper diet group were significantly decreased. Conclusion:High copper diet causes abnormality in a variety of neurobehavioral function indexes in rats, and a decrease in expression of MAP2 and GAP43 at the synaptic interface of hippocampal neurons may be involved in the process of learning and memory impairment in the neurobehavioral functions.
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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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ObjectiveTo investigate the clinical effect of perioperative Gandouling intervention in patients with Wilson’s disease (WD) complicated by splenomegaly and hypersplenism and the changes in related indices. MethodsA total of 60 WD patients with splenomegaly and hypersplenism who were hospitalized in Encephalopathy Center, The First Affiliated Hospital of Anhui University of Chinese Medicine, from July 2016 to July 2018 were enrolled and randomly divided into control group and treatment group, with 30 patients in each group. The patients in the control group were given conventional Western medicine treatment including decoppering for 4 courses (each course of treatment was 8 days), followed by splenectomy and conventional decoppering at the end of week 1 after surgery for 2 courses; the patients in the treatment group were given Gandouling in addition to the treatment in the control group. Clinical outcome and changes in 24-hour urinary copper, peripheral hemogram, liver function parameters, and portal venous flow indices were observed. The two independent samples t-test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. ResultsThe treatment group had a significantly higher overall response rate than the control group ［90% (27/30) vs 60% (18/30), χ2=443, P=0.03］. Compared with the control group at the end of two courses of treatment after surgery, the treatment group had significantly lower 24-hour urinary copper (t=41.07, P＜0.05) and levels of alanine aminotransferase and aspartate aminotransferase (t=7.29 and 6.13, both P＜0.01) and significantly higher levels of red blood cell count, platelet count, and hemoglobin (t=-5.49, -3.43, and -3.53, all P＜0.01). At the end of two courses of treatment after surgery, both groups had a reduction in portal venous flow, and the treatment group had a significantly greater improvement in portal venous flow than the control group (t=12.05, P＜0.01). ConclusionGandouling can improve the clinical outcome of WD patients with splenomegaly and hypersplenism after splenectomy.
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ObjectiveTo investigate the clinical effect of perioperative Gandouling intervention in patients with Wilson’s disease (WD) complicated by splenomegaly and hypersplenism and the changes in related indices. MethodsA total of 60 WD patients with splenomegaly and hypersplenism who were hospitalized in Encephalopathy Center, The First Affiliated Hospital of Anhui University of Chinese Medicine, from July 2016 to July 2018 were enrolled and randomly divided into control group and treatment group, with 30 patients in each group. The patients in the control group were given conventional Western medicine treatment including decoppering for 4 courses (each course of treatment was 8 days), followed by splenectomy and conventional decoppering at the end of week 1 after surgery for 2 courses; the patients in the treatment group were given Gandouling in addition to the treatment in the control group. Clinical outcome and changes in 24-hour urinary copper, peripheral hemogram, liver function parameters, and portal venous flow indices were observed. The two independent samples t-test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. ResultsThe treatment group had a significantly higher overall response rate than the control group ［90% (27/30) vs 60% (18/30), χ2=443, P=0.03］. Compared with the control group at the end of two courses of treatment after surgery, the treatment group had significantly lower 24-hour urinary copper (t=41.07, P＜0.05) and levels of alanine aminotransferase and aspartate aminotransferase (t=7.29 and 6.13, both P＜0.01) and significantly higher levels of red blood cell count, platelet count, and hemoglobin (t=-5.49, -3.43, and -3.53, all P＜0.01). At the end of two courses of treatment after surgery, both groups had a reduction in portal venous flow, and the treatment group had a significantly greater improvement in portal venous flow than the control group (t=12.05, P＜0.01). ConclusionGandouling can improve the clinical outcome of WD patients with splenomegaly and hypersplenism after splenectomy.
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The magnetic molecularly imprinted polymers (MMIPs), based on the surface of magnetic nanoparticles being modified by surface grafting, have been successfully synthesized, with dibutyl phthalate (DBP) as template molecule, ethylene glycol dimethacrylate (EGDMA) as cross-linking agent and azobisisobutyronitrile (AIBN) as initiator. Scanning electron microscopy (SEM), transmission electron microscope (TEM), and elemental analysis were employed to characterize the MMIPs. The structure and magnetic properties of the MMIPs were investigated by means of X-ray diffraction and vibrating sample magnetometer. The BET surface area shows that MMIPs is 380 m2/g and MNIPs is 324 m2/g. A series of static adsorption experiments were conducted to analyze its adsorption performance, which followed pseudo-second-order model by the kinetic analysis with correlation coefficient (R2) 0.9797, and Sips equation with correlation coefficient (R2) 0.999 by the isothermal analysis. The imprinting factors of diallyl phthalate (DAP), DBP and di-2-ethylhexyl phthalate (DEHP) were 1.53, 2.21 and 1.39 respectively, showing that MMIPs had better recognition performance for DBP. The experiment of regeneration recycles with five times showed the regeneration ability of DBP was only reduced by 12.3％.
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To establish a rat model of diabetes-associated cerebral ischemia due to qi and yin deficiency and blood stasis, and to investigate the effects of Radix Ginseng, Rhizoma Chuanxiong, Rhizoma Polygonati Odorati and Rhizoma Polygonati Sibirici Granule (Shenxiong Yujing Granule), which has the function of strengthening qi, nourishing yin, and activating blood, on proliferation, differentiation and survival of neural cells in rats with diabetes-associated cerebral ischemia.
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Objective: To explore the main characteristics of syndromes in traditional Chinese medicine (TCM) in post-stroke depression (PSD) and to provide basis for treatments with TCM herbs. Methods: According to diagnostic criteria of PSD, stroke patients and depression patients from Department of Neurology, First Affiliated Hospital, Anhui University of Traditional Chinese Medicine were assigned into cerebral stroke group (150 cases), depression group (151 cases) and PSD group (123 cases). Neuropsychological assessments and imaging and biochemical analyses were conducted. TCM syndrome differentiation for these diseases was performed. We also determined the characteristics of TCM syndromes of PSD, relative risk of the syndromes and their correlations with ages as well. Results: Scores of qi stagnation and blood stasis, liver qi depression, and transformation of fire due to qi stagnation in PSD group were significant higher than those in cerebral stroke group (P<0.05, P<0.01). In cerebral stroke group, majority of the patients displayed one syndrome, while in PSD and depression groups, the patients had three or more syndromes. Of these syndromes, the incidence rate of syndrome of liver qi depression complicated with transformation of fire due to qi stagnation or flaring of fire due to yin deficiency was high. The syndrome of liver qi depression occurred much more frequently in PSD group and depression group than in cerebral stroke group (P<0.05, P<0.01). The logistic regression analysis showed that the syndrome of qi stagnation and blood stasis had high relative risk to PSD. The syndrome of deficiency of heart and spleen was positively correlated with age in cerebral stroke group. Conclusion: The main TCM syndromes of PSD and depression are qi stagnation and blood stasis, liver qi depression, and transformation of fire due to qi stagnation. The syndrome of deficiency of heart and spleen is closely related to age among the stroke patients. The syndrome of qi stagnation and blood stasis serves as an independent risk factor for PSD. The more complicated the syndromes are, the more serious depression becomes.