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Objective To explore the characteristics and mechanism of phase separation between TAR DNA binding protein-43(TDP-43)and ubiquitin.Methods The TARDBP gene and its truncated genes were inserted into vectors to construct recombinant plasmids for expression and protein purification.The phase separation system of ubiquitin and TDP-43 was constructed in vitro.The characteristics of the droplets formed via liquid-liquid phase separation were observed by fluorescence microscopy.The plasmids of ubiquitin and TDP-43 were co-transfected into HEK293T cells to observe aggregates containing TDP-43 and ubiquitin and find out whether TDP-43 could be ubiquitinated.Results The GFP-8Ub,TDP-43 full-length(FL)and truncated proteins were purified.TDP-43 FL and C-terminal domain(CTD)proteins were able to form droplets via phase separation with ubiquitin.The droplets changed into solid-like aggregates after prolonged incubation.Insolvable aggregates containing TDP-43 and ubiquitin were formed.TDP-43 was ubiquitinated under stress conditions in HEK293T cells after being co-transfected with ubiquitin and TDP-43 recombinant plasmids.Conclusion TDP-43 undergoes co-phase separation with ubiquitin,mainly driven by the multivalent interaction between TDP-43′s CTD structural domain and ubiquitin.The droplets finally form aggregates with solid-like properties.Under stress conditions,especially when the protein homeostasis is disrupted,TDP-43 and ubiquitin form aggregates while TDP-43 is ubiquitinated.This study reveals the basic mechanism of TDP-43 co-phase separation with ubiquitin and liquid-solid transformation.
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Objective To investigate the association between serum macrophage polarization-related factors and liver fibrosis in patients with alveolar echinococcosis (AE). Methods A total of 120 patients with AE who attended Department of Hepatobiliary and Pancreatic Surgery in The Affiliated Hospital of Qinghai University from September 2018 to October 2020 were enrolled as AE group, and 33 healthy controls were enrolled as normal control group. The two groups and the patients with varying degrees of liver fibrosis were compared in terms of the levels of interleukin-6 (IL-6), interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α), and transforming growth factor-β1 (TGF-β1). Comparison of normally distributed continuous data between two groups was made by the independent samples t -test, while comparison of non-normally distributed continuous data was made by the Mann-Whitney U test or Kruskal-Wallis H test. Comparison of categorical data between groups was made by the chi-square test. Univariate and multivariate logistic regression analyses were used to investigate the association between serum macrophage polarization-related factors and liver fibrosis in patients with AE, and the receiver operating characteristic (ROC) curve was used to analyze the value of serological examination in the diagnosis of liver fibrosis in patients with AE. A Spearman correlation analysis was used to analyze the correlation of each index with HAI score and Metavir score. Results Compared with the normal control group, the AE group had significant increases in the serum levels of IL-6 [13.97 (9.64-23.62) pg/mL vs 1.30 (0.35-2.71) pg/mL, Z =-5.980, P < 0.001], TNF-α [2.26 (1.65-4.13) pg/mL vs 1.40 (1.04-2.10) pg/mL, Z =-3.114, P < 0.01], and TGF-β1 [3.64(2.71-5.72) pg/mL vs 2.91(2.20-3.35) pg/mL, Z =-2.594, P < 0.05], and increases in the serum levels of IL-6 (hazard ratio [ HR ]=2.721, 95% confidence interval [ CI ]: 1.730-4.280, P < 0.05) and TNF-α( HR =3.527, 95% CI : 1.158-10.747, P < 0.05) were independent risk factors for the onset of liver fibrosis in AE patients. The ROC curve analysis showed that hydatid IgG combined with the serum levels of IL-6 and TNF-α had a sensitivity of 88.4%, a specificity of 95.8%, and an area under the ROC curve of 0.951(95% CI : 0.937-0.964) in the diagnosis of liver fibrosis, which were significantly higher than those of IL-6, TNF-α, or hydatid IgG alone ( Z =-3.458, -4.011, and 2.379, all P < 0.05). The Spearman analysis showed that the serum levels of IL-6, TNF-α, and TGF-β1 were positively correlated with HAI score ( r =0.560, 0.644, and 0.465, all P < 0.001) and Metavir fibrosis score ( r =0.530, 0.758, and 0.567, all P < 0.001), and the serum level of IL-10 was negatively correlated with HAI score ( r =-0.232, P =0.011) and Metavir fibrosis score ( r =-0.288, P =0.001). Conclusion Macrophage polarization is often observed in patients with hepatic AE, and the levels of the macrophage polarization-related factors IL-6, TNF-α, and TGF-β1 are associated with the development and progression of liver fibrosis, which can provide certain reference information for predicting the onset of liver fibrosis.
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Objective:To analyze the risk factors for bile leakage in patients undergoing cystic echinococcosis surgery to establish a nomogram model to predict the risk of bile leakage.Methods:The clinical data were retrospectively analyzed that 203 patients with cystic echinococcosis treated in Qinghai University Affiliated Hospital from January 2015 to October 2018. Logistic regression was used to screen out independent risk factors of biliary leakage in patients undergoing cystic hydatid surgery. Then the R software was used to establish a predictive nomogram model. Bootstrap method was used to validate the nomogram model and ROC curve was used to explore the predictive efficacy of the model in predicting the bile leakage.Results:There were 54 cases of biliary leakage in 203 patients with cystic echinococcosis, the incidence was 26.1%. ALP ( OR=11.193, 95% CI 5.066-24.731), GGT ( OR=2.728, 95% CI 1.246-5.975), cyst diameter ( OR=3.491, 95% CI 1.550-7.861), hilar cyst ( OR=2.503, 95% CI 1.176-5.329) were all independent risk factors for biliary leakage of cystic echinococcosis. The consistency index was 0.835 for predicting PPC risk, and the area was 0.823 (95% CI 0.754-0.892) under the curve for predicting the risk of cystic echinococcal bile leakage. Conclusions:ALP, GGT, cyst diameter and hepatic hilar cyst are independent risk factors for cystic echinococcal bile leakage. Nomogram model can evaluate the risk of cystic echinococcal bile leakage more intuitively, with potentially high clinical application value.
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Objective:To investigate the effect of 125I particles in alveolar echinococcosis with the animal model (nude mice and Sprague Dawley rats). Methods:Twenty 10 weeks nude mice with body weight ranged from 20 to 24 g were divided into three groups. Sixteen nude mice were divided into experimental group ( n=8), puncture group ( n=4) and model group ( n=4). There was no intervention in the model group and only particle puncture needle was used in the puncture group. 125I particles were implanted in the experimental group. 14 male Sprague Dawley rats without specific pathogen, with body weight 280-320 g, 12 weeks old, were used to construct the model of hepatic alveolar echinococcosis. Then the rats were divided into intervention group ( n=10) and control group ( n=4). In the intervention group, 125I particles were pushed into the lesions. The abdomen was only open and closed in the control group. All the mice were sacrificed 45 days after intervention. The tumor size was measured. The activity of protoscolex and pathological changes of Echinococcus multilocularis in each group were observed. Results:At the timepoint of 22nd, 30th and 40th day of intervention, the largest diameter of tumor in nude mice experimental group was (10.7±5.2) mm, (10.9±5.0) mm, (8.5±4.3) mm, smaller than that in puncture group (24.5±4.4) mm, (25.4±4.1) mm, (31.4±2.8) mm and model group (22.5±7.3) mm, (25.0±5.4) mm, (26.7±6.3) mm, with statistically significant difference ( P<0.05). The number and activity of protoscoleces in experimental group were lower than those in puncture group and model group. Under the light microscope, the structure of echinococcus vesiculae and its body in the experimental group was obviously destroyed, and the cuticle and germinal layer of echinococcus vesiculae in the puncture group and the model group were normal, with multiple intact protoscoleces. The pathological changes of Sprague Dawley rats in the intervention group and the control group were basically the same as those in the nude mice model. Conclusion:The 125I particle radiation effect can kill Echinococcus multilocularis protoscoleces and inhibit the growth of alveolar echinococcosis.
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Objective:To investigate the effect of hepatitis B virus (HBV) infection in patients with hepatic alveolar echinococcoisis after surgical resection and determine the differences of liver function between patients with different HBV-DNA levels.Methods:Patients were selected from January 2014 to July 2018 in the Affiliated Hospital of Qinghai University. Twenty-eight patients with hepatitis B and hepatic alveolar echinococcoisis were included in the experimental group, and 20 patients with hepatic alveolar echinococcoisis but without hepatitis B virus were included in the control group. Based on HBV-DNA level, the experimental group was divided into low-level group (HBV-DNA level<200 IU/ml, n=6), intermediate-level group (HBV-DNA level 200-20 000 IU/ml, n=15) and high level group (HBV-DNA level>20 000 IU/ml, n=7). Comparison of complications and liver function after liver resection in two groups.Univariate and multivariate logistic regression were used to analyze the influential factors of postoperative complications in patients.Comparison of postoperative liver function indexes in patients with different HBV-DNA levels. Results:In the control group, postoperative total bilirubin 10.6(8.3, 16.9) μmol/L, direct bilirubin 5.3(3.4, 10.0) μmol/L, prothrombin time 13.6(13.0, 15.8)s, and the incidence of complications 25.0%(5/20), were better than the experimental group 12.6(8.4, 46.9) μmol/L, 6.7(3.1, 26.4) μmol/L, 15.4(13.5, 18.1)s, 78.6% (22/28), the differences were statistically significant significance (all P<0.05). Multivariate logistic analysis showed that patients with HBV infection ( OR=4.593, 95% CI: 1.128-18.708) and intraoperative blood loss ≥1 000 ml ( OR=2.200, 95% CI: 1.106-4.378) were the hepatic alveolar echinococcoisis independent risk factors for postoperative complications. There were no significant differences in total bilirubin and albumin between the three groups of patients with different HBV-DNA levels ( P>0.05). Conclusion:Patients with HBV and hepatic alveolar echinococcoisis have worse liver function and are more prone to complications after surgical resection, but there is no significant difference in liver function among patients with different HBV-DNA levels.
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Liquid biopsy is a new noninvasive detection method and is also the main molecular detection method to guide the application of precision medicine. Liquid biopsy mainly includes the measurement of circulating free DNA (cf-DNA), circulating tumor cells, and exosomes, and in particular, cf-DNA is becoming a valuable molecular detection tool, especially in the early detection and diagnosis of liver cancer and parasitic infections involving the liver. This article reviews the clinical application of cf-DNA in liver cancer and liver-related parasitic diseases.
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Objective To preliminarily study the use of indocyanine green in hepatic alveolar echinococcosis surgery.Methods The data of 13 patients with hepatic alveolar ecbinococcosis treated at the Affiliated Hospital of Qinghai University from May 2017 to May 2018 with laparotomy and intraoperative indocyanine green injection were retrospectively studied.Images were collected by the fluorescence acquisition system.Results There were 7 females and 6 males,with an average age of (37.9±14.7) years.The mean weight was (57.4±11.3) kg.Except for one patient of Han nationality,the rest were Tibetans.The lesions were not visualized but normal liver tissues emitted green fluorescence.One patient had a slightly enhanced fluorescence circle around the lesion.The fluorescence intensity of some areas were between normal liver tissues and the lesion,which were suspected to be the marginal zone (which needed to be confirmed by pathology).Three patients (23.1％,3/13) had small lesions on the liver surface,2 were not found by imaging examination,and 1 showed strip calcification on CT.No residual lesions in the liver (except for the microwave ablated lesions) were found after resection.No postoperative complications occurred in all the patients.Seven patients were followed up and no recurrence was found.Conclusions Fluorescence imaging of indocyanine green has the advantages of detecting small and residual lesions in surgery carried out for hepatic alveolar echinococcosis.More studies are needed to confirm the findings.
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Calculous cholecystitis is a common disease in clinical practice. There are various therapies for this disease, mainly cholecystectomy, and laparoscopic cholecystectomy has always been considered the gold standard for the treatment of calculous cholecystitis. However, medication, percutaneous transhepatic gallbladder drainage, and gallbladder-preserving cholecystolithotomy can be selected based on patient age, conditions, comorbidities, and preference. This article briefly describes the features of medication, interventional treatment, and surgical treatment in patients with calculous cholecystitis and points out that the optimal treatment regimen should be selected based on the features of calculous cholecystitis in each patient. This article also proposes the individualized treatment mode for calculous cholecystitis.
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HBV infection is highly prevalent in China, and patients with a high HBV-DNA load for a long time may progress to liver cirrhosis, which may lead to liver cancer. At present, surgery is still the main method for the treatment of hepatitis B-related liver cancer. Many studies have shown that HBV DNA load is an important influencing factor for the recovery of liver function after surgery, postoperative complications, recurrence of liver cancer, and the success of liver transplantation in patients with hepatitis B-related liver cancer. Its mechanism may be associated with high HBV DNA load and HBV reactivation. Maintaining a low HBV DNA load by antiviral therapy can improve the prognosis of patients with hepatitis B-related liver cancer. This article reviews the influence of HBV DNA load before and after surgery on the surgical outcome of hepatitis B-related liver cancer, in order to provide a reference for research on the treatment of patients with liver cancer.
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Objective To investigate the mechanism of Smad ubiquitination-related factor 2 (Smurf2)neddylation. Methods The Smurf2 protein level was tested by overexpression of Nedd8,while the method of immunoprecipitation(IP) and Western blotting were used to analyz Smurf2-Nedd8 modification.The GST-pulldown experiment was conducted to prove protein interactions.The protein was obtained by grinding mouse tissue and Western blotting was used to test the protein expression level.Results Over expression of Nedd8 could lead to the down regulation of the Smurf2′s protein level.Smurf1 and Smurf2 could interact in the GST-pulldown experiment. Smurf1 could enhance Smurf2-Nedd8 modification.The Smurf2′s protein level was up-regulated in mouse tissue of Smurf1 C426A.Conclusion There is some relationship and also difference between Smurf1 and Smurf2.Smurf1 can enhance Smurf2-Nedd8 modification.
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Objective To study the safety and efficacy of ultrasound-guided puncture combined with surgical resection for liquefied cavitary hepatic alveolar echinococcosis (HAE).Methods A retrospective study was conducted on 17 patients who had liquefied cavitary HAE and were treated at the Affiliated Hospital of Qinghai University from January 2014 to August 2016.Ten patients were treated with ultrasound-guided puncture combined with surgical resection (the case group),and 7 patients were treated with resection (the control group).The basic characteristics,operation time,blood loss during operation,preoperative and postoperative laboratory tests and complications were compared between the 2 groups.Results There were no significant differences between the 2 groups in basic characteristics,such as age,gender,and lesion diameter (P ＞ 0.05).The operation time and blood loss during operation showed significant differences between the 2 groups,(308.0 ± 23.0) min (389.0 ± 95.7) min and (1 360.0 ± 182.9) ml vs.(1 607.1 ± 205.0) ml,respectively (all P ＜ 0.05).The prothrombin time (PT) after day 3 of operation and alanine aminotransferase level after day 5 of operation showed significant differences between the 2 groups,(13.8 ±0.9) s vs.(15.5 ±1.7) s and (81.9 ±20.9) U/L vs.(108.1 ±29.5) U/L,respectively (all P ＜ 0.05).There was no significant difference in postoperative complications between the 2 groups.Conclusions Ultrasound-guided puncture combined with surgical resection shortened the operation time,reduced blood loss and avoided serious complications after surgery.This treatment is efficacious and safe for liquefied cavitary HAE.
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Objective To clarify the effects of HBV co-infection on liver function of patients with different types of hepatic echinococcosis.Methods We recruited 409 patients diagnosed with hepatic echinococcosis at three hospitals in western regions in China from 2014 to 2015.Venous blood was withdrawn to detect to liver function indications.ELISA was performed to detect HBsAg.We analyzed liver function in patients stratified by different types of hepatic echinococcosis with or without HBV infection.Results The hepatic echinococcosis patients infected with HBV had more severe impairment in liver functions such as reduced albumin and increased transaminase.The patients with hepatic alveolar echinococcosis were more vulnerable to HBV infection compared with those with hepatic cystic echinococcosis (38.4％ vs.86.4％, P<0.05).In addition, liver injury was more severe in patients diagnosed with alveolar hepatic echinococcosis and HBV infection compared with those diagnosed with cystic hepatic echinococcosis and HBV infection (all P<0.05).Conclusion Hepatic alveolar echinococcosis patients co-infected with HBV have worse liver injury compared with those hepatic cystic with HBV. Therefore, they deserve special attention in clinical treatment.
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Ubiquitination is one of the most major post-translational modifications playing important role in regulation of intra-cellular proteins' stability, degradation, localization and biological activity. However, these proteins are difficult to be detected due to their low abundance, short half-life. In this study, ubiquitin-binding domains (UBDs) were constructed to purify the ubiquitinated proteins from Hep3B cells. Ubiquitinated proteins and sites were detected by LC-MS/MS. A total of 1 900 potential ubiquitinated proteins were identified. Among them, 158 ubiquitinated sites were identified, belonging to 102 proteins. Bioinformatics analysis revealed that the enriched pathways of ubiquitinated proteins were closely related to tumor occurrence and development. The dysfunction of ubiquitin-proteasome has a high correlation with cell signaling and extracellular matrix changing in tumor cells.
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<p><b>BACKGROUND</b>Accumulating evidence indicates that systemic inflammation response is associated with the prognosis of various cancers. The aim of this study was to investigate the neutrophil-lymphocyte ratio (NLR), which is one of the systemic inflammation markers, in the prognosis of hepatocellular carcinoma (HCC) after treatment of transcatheter arterial chemoembolization (TACE).</p><p><b>METHODS</b>The clinical data of 178 HCC patients who received TACE were retrospectively analyzed. The optimal NLR cutoff was determined according to the receiver operating characteristic (ROC) analysis. All patients were divided into NLR-normal group and NLR-elevated group according to the cutoff, and the clinical features of these two groups were comparatively analyzed. Meanwhile, the overall survival and disease free survival (DFS) were analyzed using the Kaplan-Meier method. The risk factors of postoperative survival were investigated using univariate and multivariate Cox regression analyses.</p><p><b>RESULTS</b>The optimal NLR cutoff was defined at 1.85 and 42 (23.6%) patients had an elevated NLR (NLR>1.85). The median survival time was 9.5 months (range 1-99 months). The clinical data between the two groups were comparable, except for a-fetoprotein. Follow-up results showed that the median survival of patients with normal NLR was 17.5 months (range: 1-99 months) compared with 8 months (range: 8-68 months) of patients with elevated NLR. The 1, 3 and 5-year overall survival of patients in the NLR-normal group and NLR-elevated group were 57.3%, 44.1%, and 27.2% and 42.1%, 19.6%, and 9.5% respectively (χ(2) = 194.2, P < 0.001). Similarly, the disease free survival also has a significant difference (χ(2) = 39.3, P < 0.001). Multivariate Cox regression analysis showed that a high NLR was an independent factor affecting the survival rate of HCC after TACE (P = 0.04).</p><p><b>CONCLUSION</b>Preoperative NLR was an important prognostic factor to predict the prognosis of patients with intermediate HCC treated with TACE.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Carcinoma, Hepatocellular , Pathology , Therapeutics , Chemoembolization, Therapeutic , Liver Neoplasms , Pathology , Therapeutics , Lymphocytes , Metabolism , Physiology , Neutrophils , Metabolism , PhysiologyABSTRACT
Apoptosis action is primarily exerted at the level of mitochondira, in which Bcl-2 family of proteins play an important role in its regulation. Bcl-2 family consists of anti-apoptotic and pro-apoptotic members. The anti-apoptotic members usually exist in the outer mitochondrial membrane and inhibit cell death via interaction with pro-apoptotic counterparts BH3 domain. Pro-apoptotic members are commonly localize in the cytoplasm. A series of events occured, such as typical Bax conformational change, BAD and Bik phosphorylation as well as Bid and Bim proteolysis in response to several death stimuli. As a result, these pro-apoptotic proteins directly integrate to the outer mitochondrial membranes. Finally, mitochondrial permeability transition pore is opened, by followed the release of apoptogenic factors from the mitochondrial intermembrane space, including cytochrome c, apoptosis inducing factor(AIF) and Smac, then the activation of downstream caspases and execution of cell death.
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NEDD8 is a member of the ubiquitin-like proteins. The overall structure of NEDD8 is quite similar to ubiquitin. Covalent conjugation of NEDD8 to proteins at the post-translational level is called Neddylation. Neddylation occurs similarly to ubiquitination and need enzyme cascades involving E1, E2 and E3. Neddylation has been demonstrated to be essential to maintain the ubiquitin ligase activity of Cullin-Roc based E3 ligases. Compared with the ubiquitination which was widely studied in the past two decades, few substrates were identified for Neddylation and the physiological functions of Neddylation need further investigations. The current progress of function and regulation of protein Neddylation will be reviewed.
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Liquid biopsy is a new noninvasive detection method and is also the main molecular detection method to guide the application of precision medicine. Liquid biopsy mainly includes the measurement of circulating free DNA (cf-DNA), circulating tumor cells, and exosomes, and in particular, cf-DNA is becoming a valuable molecular detection tool, especially in the early detection and diagnosis of liver cancer and parasitic infections involving the liver. This article reviews the clinical application of cf-DNA in liver cancer and liver-related parasitic diseases.