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To enhance the DNA/RNA amplification efficiency and inhibitor tolerance of Bst DNA polymerase, four chimeric Bst DNA polymerase by fusing with a DNA-binding protein Sto7d and/or a highly hydrophobic protein Hp47 to Bst DNA polymerase large fragment. One of chimeric protein HpStBL exhibited highest inhibitor tolerance, which retained high active under 0.1 U/µL sodium heparin, 0.8 ng/µL humic acid, 2.5× SYBR Green I, 8 % (v/v) whole blood, 20 % (v/v) tissue, and 2.5 % (v/v) stool. Meanwhile, HpStBL showed highest sensitivity (93.75 %) to crude whole blood infected with the African swine fever virus. Moreover, HpStBL showed excellent reverse transcriptase activity in reverse transcription loop-mediated isothermal amplification, which could successfully detect 0.5 pg/µL severe acute respiratory syndrome coronavirus 2 RNA in the presence of 1 % (v/v) stools. The fusion of two domains with different functions to Bst DNA polymerase would be an effective strategy to improve Bst DNA polymerase performance in direct loop-mediated isothermal amplification and reverse transcription loop-mediated isothermal amplification detection, and HpStBL would be a promising DNA polymerase for direct African swine fever virus/severe acute respiratory syndrome coronavirus 2 detection due to simultaneously increased inhibitor tolerance and reverse transcriptase activity.
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Virus de la Fiebre Porcina Africana , ADN Polimerasa Dirigida por ARN , ADN Polimerasa Dirigida por ARN/metabolismo , ADN Polimerasa Dirigida por ARN/genética , Virus de la Fiebre Porcina Africana/genética , Virus de la Fiebre Porcina Africana/enzimología , Animales , Proteínas Recombinantes de Fusión/genética , SARS-CoV-2/efectos de los fármacos , SARS-CoV-2/genética , Porcinos , Técnicas de Amplificación de Ácido Nucleico/métodos , Dominios Proteicos , ADN Polimerasa Dirigida por ADN/genética , ADN Polimerasa Dirigida por ADN/metabolismo , Inhibidores de la Transcriptasa Inversa/farmacología , COVID-19/virología , ARN Viral/genéticaRESUMEN
BACKGROUND: In patients with hilar cholangiocarcinoma (HCCA), radical resection can be achieved by resection and reconstruction of the vasculature. However, whether vascular reconstruction (VR) improves long-term and short-term prognosis has not been demonstrated comprehensively. METHODS: This was a retrospective multicenter study of patients who received surgery for HCCA with or without VR. Variables associated with overall survival (OS) and recurrence-free survival (RFS) were identified based on Cox regression. Kaplan-Meier curves were used to explore the impact of VR. Restricted mean survival time (RMST) was used for comparisons of short-term survival between the groups. Patients' intraoperative and postoperative characteristics were compared. RESULTS: Totally 447 patients were enrolled. We divided these patients into 3 groups: VR with radical resections (n = 84); non-VR radical resections (n = 309) and non-radical resection (we pooled VR-nonradical and non-VR nonradical together, n = 54). Cox regression revealed that carbohydrate antigen 242 (CA242), vascular invasion, lymph node metastasis and poor differentiation were independent risk factors for OS and RFS. There was no significant difference of RMST between the VR and non-VR radical groups within 12 months after surgery (10.18 vs. 10.76 mon, P = 0.179), although the 5-year OS (P < 0.001) and RFS (P < 0.001) were worse in the VR radical group. The incidences of most complications were not significantly different, but those of bile leakage (P < 0.001) and postoperative infection (P = 0.009) were higher in the VR radical group than in the non-VR radical group. Additionally, the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) up to 7 days after surgery tended to decrease in all groups. There was no significant difference in the incidence of postoperative liver failure between the VR and non-VR radical groups. CONCLUSIONS: Radical resection can be achieved with VR to improve the survival rate without worsening short-term survival compared with resection with non-VR. After adequate assessment of the patient's general condition, VR can be considered in the resection.
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Crisis helplines provide important support for vulnerable individuals during the COVID-19 pandemic, which may also impact the helplines. We explored the challenges that the pandemic brought to Taiwan's national suicide prevention hotline and the hotline's responses. We interviewed 14 hotline workers and conducted data analysis using the framework method. The pandemic posed two new challenges to the hotline: potential service interruption and the adjustment of perceived role among hotline workers. The hotline's well-formulated response plan helped it sustain its services during the pandemic, although the workers also experienced stress and frustration resulted from role ambiguity. Our data highlighted the hotline workers' need for accurate COVID-19 information, relevant training, and timely support.
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COVID-19 , Humanos , Líneas Directas , Prevención del Suicidio , PandemiasRESUMEN
BACKGROUND/AIMS: Our previous work identified the dimethyladenosine transferase 1 homolog as a novel prognostic factor for detecting human gastric carcinoma with high sensitivity and specificity. The high expression of dimethyladenosine transferase 1 is closely associated with the occurrence and progression of gastric carcinoma. However, the underlying mechanism of dimethyladenosine transferase 1 for the occurrence and development of gastric carcinoma is not well elucidated yet. MATERIALS AND METHODS: In our present study, the biological role of dimethyladenosine transferase 1 on cell proliferation, apoptosis, and cell cycle progression in human gastric carcinoma cells was investigated through in vitro and in vivo assays by the overexpression and knockdown of dimethyladenosine transferase 1 2-way authentication method. RESULTS: We found that the overexpression of dimethyladenosine transferase 1 significantly promotes cell proliferation (P < .001) and inhibition of cell apoptosis (P < .01) in SGC-7901 cells. However, the in vivo experiment results of the knockdown dimethyladenosine transferase 1 using small interfering RNAs in the MKN-45 are just the opposite. Reverse-transcriptase polymerase chain reaction and western blotting analysis revealed that overexpressed dimethyladenosine transferase 1 in SGC-7901 cells significantly activated the AKT pathway compared to control cells. In contrast, we found that apoptosis genes such as Caspase-3 and Caspase-9 were downregulated in those cells. The xenograft nude mice model exhibited increased tumor growth (P < .01) and weight loss (P < .01), with the overexpression of dimethyladenosine transferase 1 homolog in the SGC-7901 cells. These results have been further confirmed through backward verification in dimethyladenosine transferase 1 knockdown cells. CONCLUSIONS: Taken together, our results indicated that the dimethyladenosine transferase 1 plays a crucial role in stimulating cancer cell proliferation and contributes to apoptosis resistance in human gastric carcinoma. Meanwhile, it provides a potential therapeutic target for gastric carcinoma treatment and is worthy of further studies.
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Carcinoma , Neoplasias Gástricas , Animales , Ratones , Humanos , Proteínas Proto-Oncogénicas c-akt/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transferasas/genética , Transferasas/metabolismo , Ratones Desnudos , Línea Celular Tumoral , Apoptosis/genética , Carcinoma/patología , Neoplasias Gástricas/patología , Proliferación Celular/genética , Regulación Neoplásica de la Expresión GénicaRESUMEN
It has been reported that cadherin 6 (CDH6) upregulation is associated with enhanced epithelial-to-mesenchymal transition (EMT) in several types of solid tumor cells. The current study aimed to explore the effect of CDH6 on the migration and invasion of stomach adenocarcinoma (STAD) cells, the transcription factors involved in CDH6 dysregulation and their effect on mitochondrial fission. Bioinformatics analysis was performed using data extracted from the Genotype-Tissue Expression Project, the Cancer Genome Atlas and Kaplan-Meier plotter. AGS and HGC27 cells were used to establish an in vitro STAD cell model. The results showed that higher CDH6 expression was associated with significantly shorter overall survival in patients with STAD. In addition, CDH6 overexpression promoted wound healing, enhanced the invasion ability of tumor cells and increased mitochondrial fission. Glioma-associated oncogene family zinc finger 2 (GLI2) could bind to the CDH6 promoter and activate its transcription. Fluorescent labeling also showed that GLI2 overexpression promoted mitochondrial fission. However, CDH6 silencing significantly reduced mitochondrial fragmentation. Besides, GLI2 overexpression notably upregulated phosphorylated-focal adhesion kinase and dynamin-related protein 1. However, the above effects were largely abrogated by CDH6 knockdown. In conclusion, the present study suggested that the novel GLI2/CDH6 axis could enhance the migration, invasion and mitochondrial fission of STAD cells.
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Adenocarcinoma , Neoplasias Gástricas , Humanos , Adenocarcinoma/genética , Cadherinas/metabolismo , Línea Celular Tumoral , Movimiento Celular/genética , Transición Epitelial-Mesenquimal/genética , Regulación Neoplásica de la Expresión Génica , Dinámicas Mitocondriales , Proteínas Nucleares/metabolismo , Neoplasias Gástricas/genética , Neoplasias Gástricas/patología , Proteína Gli2 con Dedos de Zinc/metabolismoRESUMEN
We investigated the impact of the COVID-19 pandemic on call volumes and call characteristics using data from a national crisis helpline. Data were extracted for 215,066 calls to Taiwan's national suicide prevention hotline (January 2018-May 2020). We used negative binomial regression to investigate changes in the weekly number of calls during the early period of the COVID-19 outbreak (January 21, 2020-May 25, 2020), relative to that expected according to the pre-pandemic trend. The call characteristics during the pandemic period (February 18, 2020-May 31, 2020) were compared between COVID-19 related vs unrelated calls. Higher-than-expected call volumes started from the 6th week of the pandemic and reached a peak in the 14th week, which was 38% (rate ratio = 1.38, 95% confidence interval 1.26-1.51) higher than that expected based on the pre-pandemic trend. The higher-than-expected call volumes were mainly attributable to higher-than-expected calls from non-suicidal and male callers. Calls in which COVID-19 was mentioned (13.2%) were more likely to be from male and first-time callers, occur outside 12 am-6 am, last less than 5 min, and were less likely to be from callers who had previous suicide attempts, recent suicidal ideation or suicide plans or actions than COVID-19 unrelated calls. Callers who made COVID-19 related calls were more likely to request information than other callers. Crisis helplines should strategically adapt to the increased need and callers' specific concerns related to the outbreak.
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Background: The COVID-19 pandemic and its consequences may affect population mental health and suicide risk. Aims: To explore the experiences among suicidal individuals who made calls to a suicide prevention hotline and to identify factors and psychological responses that may influence suicide risk. Method: We identified 60 eligible recorded calls to Taiwan's suicide prevention hotline (January 23, 2020-May 31, 2020) and analyzed the transcripts using a framework analysis. Results: We identified three themes: (a) effects of the COVID-19 pandemic on society (impacts on local economies, the fear of contagion, and disruptions caused by outbreak control measures); (b) stress experienced by callers, including increased challenges (financial burden, restricted freedom of movement, interpersonal conflicts, feelings of uncertainty, and education/career interruption) and reduced support (reduced access to health services and social support); and (c) the callers' psychological responses to stress, including anxiety, sleep disturbance, depression, loneliness, hopelessness, and entrapment, which may increase suicide risk. Limitations: Only the experiences among those who sought help by calling the hotline during the early months of the pandemic in 2020 were explored. Conclusion: Our findings revealed the potential process underlying the impact of the COVID-19 pandemic on suicide risk and have implications for prevention and intervention strategies.
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COVID-19 , Humanos , Pandemias , Líneas Directas , Prevención del Suicidio , Ideación SuicidaRESUMEN
BACKGROUND/AIMS: Human leukocyte antigen-G (HLA-G) plays an important role in inhibiting natural killer (NK) cell function and promoting immune escape. However, the specific mechanism of HLA-G on NK in gastric cancer (GC) remains not well understood. This study investigated the expression of HLA-G in GC and the role of HLA-G-effected NK cells in GC progression. METHODS: HLA-G expression in GC tissues obtained from 49 patients with GC was analyzed by immunohistochemistry and western blot. The number of tumor-infiltrating NK cells and the expression of their surface receptors were analyzed by immunohistochemistry and flow cytometry, respectively. The effect of HLA-G on NK cell proliferation was examined by Cell Counting Kit-8 (CCK8) assay. LDH release assay was used to evaluate the effect of HLA-G on the cytotoxic activity of NK cells, and the levels of IFN-γ and TNF-α in the co-cultured supernatant were detected by ELISA. Mice bearing a xenograft tumor model were used to examine the effect of HLA-G on the anti-tumor effect of NK cells. RESULTS: HLA-G positive expression was detected in most of the GC tissues, and was correlated with the adverse prognosis of the disease. The expression of HLA-G was negatively associated with the number of tumor-infiltrating NK cells. Furthermore, GC cell lines with overexpressed HLA-G revealed their ability to inhibit the cell proliferation and cytotoxic activity of NK-92MI cells, and reduce the secretion of IFN-γ and TNF-α through immunoglobulin-like transcript 2 (ILT2). Finally, this in vivo experiment was able to prove that HLA-G can inhibit the anti-tumor effect of NK cells through ILT2. CONCLUSION: The expression of HLA-G was strongly correlated with the adverse prognosis of GC. The reason may be that it inhibits the proliferation and cytotoxic activity of infiltrating NK cells through ILT2.
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Antígenos CD/metabolismo , Antígenos HLA-G/metabolismo , Células Asesinas Naturales/inmunología , Receptor Leucocitario Tipo Inmunoglobulina B1/metabolismo , Neoplasias Gástricas/patología , Anciano , Animales , Antígenos CD/genética , Línea Celular Tumoral , Proliferación Celular , Técnicas de Cocultivo , Supervivencia sin Enfermedad , Femenino , Antígenos HLA-G/genética , Humanos , Interferón gamma/análisis , Interferón gamma/metabolismo , Células Asesinas Naturales/metabolismo , Receptor Leucocitario Tipo Inmunoglobulina B1/genética , Activación de Linfocitos/inmunología , Masculino , Ratones Endogámicos NOD , Ratones SCID , Persona de Mediana Edad , Pronóstico , Receptores KIR2DL4/genética , Receptores KIR2DL4/metabolismo , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/mortalidad , Trasplante Heterólogo , Factor de Necrosis Tumoral alfa/análisis , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
This study seeks to clarify and determine the fundamental properties of N-terminal domain of high molecular weight glutenin subunits (HMW-GS) 1Dx5 (1Dx5-N). 1Dx5-N was expressed in E. coli and its solubility was measured by spectrophotometry. Effects of edible salts (NaCl, Na2CO3), disulfide bond reductant dithiothreitol (DTT) and hydrophobic interactions of denaturant sodium dodecyl sulfonate (SDS) on 1Dx5-N polymer were investigated by native polyacrylamide gelelectrophoresis (PAGE), nonreducing/reducing SDS-PAGE, intrinsic fluorescence, size exclusion chromatography (SEC), dynamic light scattering (DLS) and circular dichroism (CD). Results showed that 1Dx5-N formed a soluble aggregate in aqueous solutions by native-PAGE, clarifying the role of N-terminal of HMW-GS in the insolubility of the whole subunits. Meanwhile, the hydrophobic interaction was more potent in promoting the aggregation of 1Dx5-N in aqueous solutions from the results of SEC, DLS and CD. Edible salts, NaCl and Na2CO3, could improve the polymer formation of 1Dx5-N through disulfide bonds. Moreover, Na2CO3 at high concentrations (>200mM) greatly favored polymer formation by disulfide bonds, and it induced other types of cross-links between amino acids in 1Dx5-N according to nonreducing/reducing SDS-PAGE and fluorescence spectrum. Moreover, the formation of covalent bonds was reinforced by hydrophobic interactions between 1Dx5-N. Therefore, these results provide much novel information on the N-terminal domain of HMW-GS to facilitate the understanding of its functional properties in wheat flour.
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BACKGROUND: Hyperphosphatemia, secondary hyperparathyroidism (SHPT) and anemia are common secondary complications in hemodialysis patients with end-stage renal disease (ESRD). Compared with conventional hemodialysis (CHD), short daily hemodialysis (sDHD) has been found to be more effective in patients with ESRD. The objective of this study was to determine whether sDHD could improve hyperphosphatemia, SHPT and anemia in patients with ESRD. METHODS: Twenty-seven patients (11 women and 16 men, 46.8 ± 13.4 years old) were switched from CHD to sDHD. All hematologic parameters were measured prior to the switch (baseline), at 3 months after the switch (sDHD(1)) and at 6 months after the switch (sDHD(2)). RESULTS: The serum phosphate decreased from 2.54 ± 0.32 mmol/L at baseline to 2.15 ± 0.36 mmol/L (p < 0.001) at sDHD(1) and 1.97 ± 0.33 mmol/L (p < 0.001) at sDHD(2). Calcium-phosphate product decreased from 5.18 ± 1.24 mmol(2)/L(2) at baseline to 4.20 ± 0.71 mmol(2)/L(2) (p < 0.001) at sDHD(1) and 4.02 ± 0.83 mmol(2)/L(2) (p < 0.001) at sDHD(2). The serum PTH levels decreased from 223.9 ± 124.7 pmol/L at baseline to 196.3 ± 101.3 pmol/L (p < 0.05) at sDHD(2). The hemoglobin concentration increased significantly from CHD to sDHD. However, the requirement for erythropoietin (EPO) dose decreased from 6847.8 ± 1057.3 u/week at baseline to 5869.6 ± 1094.6 u/week (p < 0.05) at sDHD(2). CONCLUSIONS: sDHD may decrease serum phosphate, calcium-phosphate product and PTH, increase hemoglobin levels and decrease exogenous EPO dose requirements compared with CHD in hemodialysis patients.
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Anemia/prevención & control , Hiperparatiroidismo Secundario/prevención & control , Hiperfosfatemia/prevención & control , Fallo Renal Crónico/terapia , Diálisis Renal/métodos , Adulto , Anciano , Anemia/sangre , Anemia/etiología , Fosfatos de Calcio/sangre , Eritropoyetina/administración & dosificación , Femenino , Estado de Salud , Hemoglobinas/metabolismo , Humanos , Hiperparatiroidismo Secundario/sangre , Hiperparatiroidismo Secundario/etiología , Hiperfosfatemia/sangre , Hiperfosfatemia/etiología , Fallo Renal Crónico/sangre , Fallo Renal Crónico/complicaciones , Masculino , Salud Mental , Persona de Mediana Edad , Hormona Paratiroidea/sangre , Calidad de Vida , Diálisis Renal/efectos adversosRESUMEN
The asymmetric unit of the title compound, [CdCl(2)(C(7)H(10)N(2)O(2)S)(2)], contains two complex molecules with similar configurations. The Cd(II) atoms are each six-coordinated by two thiazole N and two carbonyl O atoms from the 2-(2-amino-1,3-thiazol-4-yl)acetate ligand, and by two Cl(-) anions in a distorted octa-hedral geometry. In the crystal, intra- and inter-molecular N-Hâ¯Cl hydrogen bonds create parallel chains along [1-10]. C-Hâ¯Cl inter-actions also occur.
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The growth factor progranulin (PGRN) has been implicated in embryonic development, tissue repair, tumorigenesis, and inflammation, but its receptors remain unidentified. We report that PGRN bound directly to tumor necrosis factor receptors (TNFRs) and disturbed the TNFα-TNFR interaction. PGRN-deficient mice were susceptible to collagen-induced arthritis, and administration of PGRN reversed inflammatory arthritis. Atsttrin, an engineered protein composed of three PGRN fragments, exhibited selective TNFR binding. PGRN and Atsttrin prevented inflammation in multiple arthritis mouse models and inhibited TNFα-activated intracellular signaling. Collectively, these findings demonstrate that PGRN is a ligand of TNFR, an antagonist of TNFα signaling, and plays a critical role in the pathogenesis of inflammatory arthritis in mice. They also suggest new potential therapeutic interventions for various TNFα-mediated pathologies and conditions, including rheumatoid arthritis.
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Artritis Experimental/tratamiento farmacológico , Artritis Experimental/inmunología , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Receptores Tipo II del Factor de Necrosis Tumoral/metabolismo , Receptores Tipo I de Factores de Necrosis Tumoral/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Adolescente , Adulto , Anciano , Animales , Antiinflamatorios no Esteroideos/metabolismo , Antiinflamatorios no Esteroideos/farmacología , Antiinflamatorios no Esteroideos/uso terapéutico , Artritis Experimental/patología , Artritis Experimental/fisiopatología , Cartílago Articular/metabolismo , Cartílago Articular/patología , Femenino , Granulinas , Humanos , Péptidos y Proteínas de Señalización Intercelular/química , Péptidos y Proteínas de Señalización Intercelular/genética , Péptidos y Proteínas de Señalización Intercelular/uso terapéutico , Ligandos , Masculino , Ratones , Ratones Endogámicos , Ratones Noqueados , Ratones Transgénicos , Persona de Mediana Edad , Progranulinas , Dominios y Motivos de Interacción de Proteínas , Receptores Tipo I de Factores de Necrosis Tumoral/genética , Receptores Tipo II del Factor de Necrosis Tumoral/genética , Proteínas Recombinantes de Fusión/metabolismo , Proteínas Recombinantes de Fusión/farmacología , Proteínas Recombinantes de Fusión/uso terapéutico , Proteínas Recombinantes/uso terapéutico , Transducción de Señal , Linfocitos T Reguladores/inmunología , Linfocitos T Reguladores/fisiología , Adulto JovenRESUMEN
OBJECTIVE: To prepare a novel MRI targeted contrast agent Gd-DTPA-Granzyme B monoclonal antibody (mAb) and to test its reaction conditions. METHODS: The Granzyme B mAb was coupled with DTPA,and then conjugated with Gd. The Gd-DTPA antibody was characterized using MALDI-TOF-MS. Cytotoxicity test was performed with MTT assay, and immune activation was examined with immunohistochemistry. RESULT: MALDI-TOF-MS demonstrated that the molecular weight shifted from granzyme B mAb (133986) to Gd-DTPA-GB mAb (139736), which indicated the conjugation of the antibody with Gd-DTPA. The molar ratio of Gd per IgG molecule was about 20. MTT assay showed that Gd, DTPA, Gd-DTPA and Gd-DTPA-GB mAb groups did not make an impact on cell viability, and there were no significant differences among 4 groups (P>0.05). Immunohistochemistry results showed that compared with the positive control group the targeted contrast agent had a high immune activity. CONCLUSION: The novel contrast agent Gd-DTPA-Granzyme B mAb prepared in this study keeps a good immune activity and has no significant cytotoxicity.
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Anticuerpos Monoclonales/química , Medios de Contraste/química , Gadolinio DTPA/química , Granzimas/inmunología , Células Cultivadas , Medios de Contraste/toxicidad , Imagen por Resonancia Magnética , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
BACKGROUND: Hepatic ischemia-reperfusion (I/R) injury occurs in many clinical procedures. The molecular mechanisms responsible for hepatic I/R injury however remain unknown. Sphingolipids, in particular ceramide, play a role in stress and death receptor-induced hepatocellular death, contributing to the progression of several liver diseases including liver I/R injury. In order to further define the role of sphingolipids in hepatic I/R, systemic analysis of sphingolipids after reperfusion is necessary. METHODS: We investigated the lipidomic changes of sphingolipids in a rat model of warm hepatic I/R injury, by delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry (DE MALDI-TOF-MS). RESULTS: The total amounts of ceramide and sphingomyelin and the intensity of most kinds of sphingolipids, mainly sphingomyelin, significantly increased at 1 hour after reperfusion (P < 0.05) and reached peaks at 6 hours after reperfusion (P < 0.01) compared to controls. Six new forms of ceramide and sphingomyelins appeared 6 hours after reperfusion, they were (m/z) 537.8, 555.7, 567.7, 583.8, 683.5 and 731.4 respectively. A ceramide-monohexoside (m/z) 804.4 (CMH(d18:1C22:1+Na)(+)) also increased after reperfusion and correlated with extent of liver injury after reperfursion. CONCLUSIONS: Three main forms of sphingolipids, ceramide, sphingomyelin and ceramide-monohexoside, are related to hepatic I/R injury and provide a new perspective in understanding the mechanisms responsible for hepatic I/R injury.