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Adulteration by Bacopa monnieri (BM) in Portulaca oleracea (PO) plants frequently occurs; it decreases the efficacy of traditional Chinese medicine (TCM) and leads to fraud in the herbal marketplace. In this study, a diagnostic PCR assay was established for the rapid authentication of PO and BM in the herbal market. The sequence divergences in internal transcribed spacer 2 (ITS2) between PO and its adulterant species were used to design diagnostic PCR primers. The specific designed primer sets were evaluated and show that the diagnostic PCR assay can be used to verify the authenticity of PO and BM. The detection limits of the primer set for PO and BM identification were 10 pg and 1 pg, respectively. The reactivity of diagnostic PCR was 0.1% PO genomic DNA and 0.01% BM genomic DNA in the test sample during DNA amplification. In addition, multiplex PCR (mPCR) for PO and BM identification was also established. The samples were more susceptible to the effect of steaming in authentication by singleplex PCR and mPCR than boiling and drying treatment. Furthermore, commercial samples from the market were used to demonstrate the applicability of the developed diagnostic PCR for PO authentication and diagnose BM adulteration, and the investigation found that approximately 72.2% (13/18) of PO plants in the herbal market were adulterated. In conclusion, the diagnostic PCR assay was successfully developed and its specificity, sensitivity and reactivity for PO and BM authentication were proven. These developed PCR-based molecular methods can be applied as an identification tool for PO authenticity and can be practically applied for inspection of BM adulteration in the herbal market in the future.
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Plantas Medicinales , Portulaca , Plantas Medicinales/genética , Portulaca/genética , Reacción en Cadena de la Polimerasa Multiplex , ADN Espaciador Ribosómico/genética , ADN de Plantas/análisis , ADN de Plantas/genéticaRESUMEN
Portulaca oleracea (PO) is a commonly known medicinal crop that is an important ingredient for traditional Chinese medicine (TCM) due to its use as a vegetable in the diet. PO has been recorded to be frequently adulterated by other related species in the market of herbal plants, distorting the PO plant identity. Thus, identification of the botanical origin of PO is a crucial step before pharmaceutical or functional food application. In this research, a quick assay named "loop-mediated isothermal amplification (LAMP)" was built for the specific and sensitive authentication of PO DNA. On the basis of the divergences in the internal transcribed spacer 2 (ITS2) sequence between PO and its adulterant species, the LAMP primers were designed and verified their specificity, sensitivity, and application for the PO DNA authentication. The detection limit of the LAMP assay for PO DNA identification specifically was 100 fg under isothermal conditions at 63 °C for 30 min. In addition, different heat-processed PO samples can be applied for use in PO authentication in the LAMP assay. These samples of PO were more susceptible to the effect of steaming in authentication by PCR than boiling and drying treatment. Furthermore, commercial PO samples pursued from herbal markets were used to display their applicability of the developed LAMP analysis for PO postharvest authentication, and the investigation found that approximately 68.4% of PO specimens in the marketplace of herbal remedies were adulterated. In summary, the specific, sensitive, and rapid LAMP assay for PO authentication was first successfully developed herein, and its practical application for the inspection of adulteration in PO samples from the herbal market was shown. This LAMP assay created in this study will be useful to authenticate the botanical origin of PO and its commercial products.
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Plantas Medicinales , Portulaca , Portulaca/genética , Plantas Medicinales/genética , Técnicas de Amplificación de Ácido Nucleico , Reacción en Cadena de la Polimerasa , Cartilla de ADN/genética , ADN , Sensibilidad y EspecificidadRESUMEN
Homogenously dispersing single-walled carbon nanotubes (SWNTs) in solvents has been one critical step towards exploiting their exceptional properties in high-performance components. However, the solubility of SWNTs is severely limited by the inert tube surfaces and strong tube-tube van der Waals attractions. Starting with carbon nanotubides, i.e., negatively charged SWNTs reduced by alkali metals, we herein propose a sonication-free approach to prepare an aqueous dispersion of SWNTs. The approach combines the spontaneous dissolution of nanotubides in polar aprotic solvents with polyvinylpyrrolidone wrapping and dialysis in deionized H2O, which results in well-dispersed, neutralized SWNTs. The gelation of concentrated SWNT dispersion leads to the formation of hydrogels, which is subsequently transformed into SWNT aerogels through lyophilization. The prepared SWNT aerogels exhibit high-mass-sorption capacities for organic solvent absorption, paving the way towards harvesting the extraordinary properties of SWNTs.
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Accumulating evidence has indicated an increased risk of acute pancreatitis in individuals with inflammatory bowel disease (IBD); however, the establishment of a clear and direct causal connection between IBD and acute pancreatitis remains uncertain. Utilizing genetic data from publicly accessible genome-wide association studies (GWAS), we conducted a 2-sample MR analysis to identify the associations between IBD, ulcerative colitis (UC), Crohn disease (CD), and acute pancreatitis risk. Rigorous quality control steps ensured the selection of eligible single nucleotide polymorphisms (SNPs) with strong associations to IBD. The primary estimation used the inverse-variance weighted method. We also assessed heterogeneity, potential pleiotropy, and conducted sensitivity analyses. The direction of causality was confirmed using the Steiger test. The MR analysis showed that IBD increased the risk of acute pancreatitis (IVW: OR = 1.032, 95% CI: 1.006-1.06, P = .015). Among the subgroup of IBD, CD (IVW: OR = 1.034, 95% CI: 1.008-1.06, P = .007) indicates a significant increase in the risk of acute pancreatitis compared to UC (IVW: OR = 1.02, 95% CI: 0.99-1.051, P = .189). The MR analysis assessing the association between CD and acute pancreatitis showed no evidence of heterogeneity or horizontal pleiotropy. Likewise, the leave-one-out (LOO) method indicated no significant influence of any individual SNP on the overall findings. In addition, the Steiger direction test revealed that CD was the cause for increased risk of acute pancreatitis, but not vice versa. In summary, this research pioneers in proposing a causal relationship between CD and acute pancreatitis among the European population.
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Colitis Ulcerosa , Enfermedad de Crohn , Estudio de Asociación del Genoma Completo , Análisis de la Aleatorización Mendeliana , Pancreatitis , Polimorfismo de Nucleótido Simple , Humanos , Colitis Ulcerosa/genética , Colitis Ulcerosa/epidemiología , Colitis Ulcerosa/complicaciones , Enfermedad de Crohn/genética , Enfermedad de Crohn/epidemiología , Pancreatitis/genética , Pancreatitis/epidemiología , Pancreatitis/etiología , Predisposición Genética a la Enfermedad , Factores de Riesgo , Enfermedad AgudaRESUMEN
Background: Accumulating evidence has shown that patients with inflammatory bowel disease (IBD) have liver function abnormalities and are susceptible to liver diseases. However, the existence of a causal relationship between IBD and liver function or disease remains unclear. Methods: A two-sample Mendelian randomization (MR) analysis was performed using genetic associations from publicly available genome-wide association studies (GWAS). These associations encompass ulcerative colitis (UC), Crohn's disease (CD), liver function traits, and liver disease phenotypes. The liver function traits comprised hepatic biochemistries, percent liver fat, and liver iron content from the UK Biobank. Furthermore, the liver disease phenotypes included cholelithiasis, non-alcoholic fatty liver disease (NAFLD), primary sclerosing cholangitis (PSC), and primary biliary cholangitis (PBC) in cohorts of European ancestry. The primary estimation used the inverse-variance weighted method, with GWAS of C-reactive protein (CRP) in the UK Biobank serving as a positive control outcome. Results: Genetically predicted UC is causally associated with decreased levels of albumin (ALB) and liver iron content, while genetically predicted CD is causally associated with increased levels of alkaline phosphatase (ALP). Moreover, genetically predicted UC or CD increases the risk of PSC, and CD increases the risk of PBC. Neither UC nor CD causally increases the risk of cholelithiasis and NAFLD. Conclusion: UC affects the levels of ALB and liver iron content, while CD affects the levels of ALP. Both UC and CD increase the risk of PSC, and CD increases the risk of PBC.
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Recent studies have indicated strong connections between epigenetic modulation and secondary metabolites in plants. It is vital to understand the roles of epigenetics in the production of secondary metabolites. In this study, the inhibitor of DNA methylation 5-azacytidine (5-Az) was used on the hairy roots of the medicinal plant Salvia miltiorrhiza to investigate its effect on secondary metabolite production, gene expression, methylation levels in genomic DNA and promoter regions. Our results showed that the contents of tanshinones in S. miltiorrhiza hairy roots increased by 1.5-5 times, and some genes in the biosynthesis pathway showed an upward trend. According to our NGS analysis, the methylation pattern in the promotor of the gene encoding copalyl diphosphate synthase (CPS) was altered, and 51 out of 145 cytosines were demethylated during 5-Az treatment. A total of 36 putative transcription factors (TFs) binding cites were identified in these demethylation sites. Among these TFs binding cites, cis-regulatory elements for the binding of NF-Y and MYB were frequently found in our results. This is the first report to demonstrate a possible mechanism of DNA methylation participating in tanshinone biosynthesis in S. miltiorrhiza hairy roots by modulating the CPS promoter and TFs binding sites.
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Salvia miltiorrhiza , Abietanos , Azacitidina/metabolismo , Azacitidina/farmacología , Epigénesis Genética , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/genética , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Salvia miltiorrhiza/metabolismoRESUMEN
Background: In Taiwan, the aerial part of Adenostemma lavenia (Al) is used in the form of herbal tea or in a folk remedy primarily to mitigate inflammatory conditions in the lungs and liver. Due to the excellent health benefits of Al against inflammation, it has become increasingly crucial and in great demand during the COVID-19 pandemic. However, Al has been found to be adulterated with Wedelia biflora, Sigesbeckia orientalis, and/or Wedelia chinensis because of similarities in appearance and vernacular names. Methods: This study aimed to develop a PCR-RFLP DNA molecular method for the authentication of Al. The restriction enzyme BsrI was used according to the sequencing and alignment results of PCR products in the ITS2 regions of Al and its adulterants. Gel electrophoresis resulted in the clear separation of Al and its adulterants into two distinct categories. Results: In conclusion, the PCR-RFLP authentication method developed herein provides an easy, rapid, and accurate method to distinguish Al from its adulterants to assure user health and safety.
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Plantas Medicinales , Reacción en Cadena de la Polimerasa , COVID-19 , ADN de Plantas/genética , Pandemias , Plantas Medicinales/clasificación , Plantas Medicinales/genética , Polimorfismo de Longitud del Fragmento de RestricciónRESUMEN
AIMS: To design a model to predict the early prognosis of patients with traumatic brain injury (TBI) based on parameters that can be quickly obtained in emergency conditions from medical history, physical examination, and supplementary examinations. METHODS: The medical records of TBI patients who were hospitalized in two medical institutions between June 2015 and June 2021 were collected and analyzed. Patients were divided into the training set, validation set, and testing set. The possible predictive indicators were screened after analyzing the data of patients in the training set. Then prediction models were found based on the possible predictive indicators in the training set. Data of patients in the validation set and the testing set was provided to validate the predictive values of the models. RESULTS: Age, Glasgow coma scale score, Apolipoprotein E genotype, damage area, serum C-reactive protein, and interleukin-8 (IL-8) levels, and Marshall computed tomography score were found associated with early prognosis of TBI patients. The accuracy of the early prognosis prediction model (EPPM) was 80%, and the sensitivity and specificity of the EPPM were 78.8% and 80.8% in the training set. The accuracy of the EPPM was 79%, and the sensitivity and specificity of the EPPM were 66.7% and 86.2% in the validation set. The accuracy of the early EPPM was 69.1%, and the sensitivity and specificity of the EPPM were 67.9% and 77.8% in the testing set. CONCLUSION: Prediction models integrating general information, clinical manifestations, and auxiliary examination results may provide a reliable and rapid method to evaluate and predict the early prognosis of TBI patients.
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Lesiones Traumáticas del Encéfalo , Humanos , Escala de Coma de Glasgow , Lesiones Traumáticas del Encéfalo/diagnóstico , Pronóstico , Tomografía Computarizada por Rayos XRESUMEN
OBJECTIVE: To investigate the influence of apolipoprotein E (APOE) gene polymorphism on regional cerebral oxygen saturation (rScO2) and quantitative electroencephalogram (QEEG) at the early phase of adult traumatic brain injury (TBI). METHODS: clinical data of TBI patients who were admitted to the neurosurgery intensive care unit (NICU) were retrospectively evaluated and studied, and data of healthy volunteers were recruited as control. The APOE genotypes were genotyped by quantitative fluorescent polymerase chain reaction (QF-PCR). The rScO2 and brainelectricalactivityof all the participants involved in this research were measured by near-infrared spectroscopy (NIRS) and QEEG respectively. RESULTS: The average rScO2 of TBI patients was significantly lower than that of the normal controls (P < 0.0001). And the EEG of the TBI patients has showed more irregular slow-wave activities than that of the normal controls. Furthermore, the above changes were more significant in the APOE ε4 carriers in the early stage of TBI patients. CONCLUSIONS: The APOE ε4 allele may be associated with poor rScO2 and more slow-wave activities at the early stage of TBI. SIGNIFICANCE: To clarify the effect of APOE gene polymorphism on the condition of patients with TBI may be helpful for the design and management of individualized treatment programs.
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Apolipoproteína E4 , Lesiones Traumáticas del Encéfalo , Adulto , Apolipoproteína E4/genética , Apolipoproteínas E/genética , Lesiones Traumáticas del Encéfalo/diagnóstico , Lesiones Traumáticas del Encéfalo/genética , Electroencefalografía , Genotipo , Humanos , Saturación de Oxígeno , Polimorfismo Genético/genética , Estudios RetrospectivosRESUMEN
AIM: To establish an artery and venous sinus occlusion image score (AVOIS) which is compatible in both cerebral arteries and venous system diseases. METHODS: A total of 188 consecutive patients with the final diagnosis of anterior circulation infarct (ACI) and 56 consecutive patients with cerebral venous and sinus thrombosis (CVST) were retrospectively studied. The AVOIS was developed based on the severity of occlusive changes of main intracranial arteries and venous sinuses (present = 0, partial occlusion = 1, absent = 2), and divided into four groups (CVST group: 0, 1-5, 6-10, >10. ACI group: 0, 1-5, 6-10, >10) arbitrarily. A receiver operating characteristic (ROC) curve was applied to discover the sensitivity and specificity of AVOIS. The National Institutes of Health Stroke Scale (NIHSS), Clot Burden Score (CBS) were set as the reference. Logistic regression models were developed to adjust for baseline clinical variables and AVOIS. Length of hospital stay (LOS) was also evaluated using the Kaplan-Meier estimator. RESULTS: For the CVST group, a positive correlation between AVOIS and NIHSS was discovered (Spearman's ρ = 0.54, p < 0.001). For the ACI group, ROC showed relatively high sensitivity (84.8%) and specificity (81.8%). Besides, the probability of time to discharge was significantly different among the AVOIS subgroups as well (p < 0.001). CONCLUSION: The AVOIS can be used to evaluate the treatment of patients with acute stroke caused by cerebral venous sinus thrombosis and anterior circulation large vessel occlusion. It is a reliable and convenient method that may help prompt prognosis and guide the treatment of individual patients.
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Nanofat grafting is a fat transfer procedure that uses a thin needle to smooth out wrinkles, thereby achieving the goal of skin rejuvenation. The Luer-Lok connector is one of the most common methods for obtaining Nanofat. In the present study, we compared three different Luer-Lok connectors (2.0 mm, 1.5 mm and 1.1 mm in diameter) in terms of their impact on the viability of adipose-derived stem cells (ADSCs) to determine the optimal size of the connector for efficient Nanofat grafting. We observed that a smaller diameter of the Luer-Lok connector created a higher mechanical shear force, which broke more fat cells during the emulsifying procedure, thereby reducing the viability of ADSCs from the stromal vascular fraction (SVF). Nanofat obtained from the 2-mm Luer-Lok connector had a better effect on skin rejuvenation than the 1.5-mm and 1.1-mm connectors. Therefore, this study presents an advance in the simple procedure of preparing Nanofat based on a previous technique and provides evidence that a procedure associated with less trauma may be a better choice.
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Tejido Adiposo/citología , Trasplante de Células Madre/instrumentación , Jeringas , Adulto , Células Cultivadas , Diseño de Equipo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Rejuvenecimiento , Envejecimiento de la Piel , Trasplante Autólogo , Factor A de Crecimiento Endotelial Vascular , Adulto JovenRESUMEN
BACKGROUND: Obesity is caused by excessive accumulation of body fat and is closely related to complex metabolic diseases. Raspberry ketone (RK), a major aromatic compound in red raspberry, was recently reported to possess anti-obesity effects. However, its mechanisms are unclear. AIM: Adipogenesis plays a critical role in obesity and, therefore, this study aimed to investigate the effect and mechanisms of action of RK on adipogenesis in 3T3-L1 preadipocytes. MATERIALS AND METHODS: 3T3-L1 preadipocytes were differentiated in medium containing insulin, dexamethasone, and 1-methyl-3-isobutylxanthine. Adipocyte lipid contents were determined using oil-red O staining while adipogenic transcription factor and lipogenic protein expressions were determined using western blotting. RESULTS: RK (300-400µM) strongly inhibited lipid accumulation during 3T3-L1 preadipocyte differentiation into adipocytes. RK reduced the CCAAT/enhancer-binding protein-α (C/EBP-α), peroxisome proliferation-activated receptor-γ (PPAR-γ), fatty acid synthase (FAS), and fatty acid-binding protein 4 (FABP4) expressions and increased heme oxygenase-1 (HO-1), Wnt10b, and ß-catenin expressions in 3T3-L1 adipocytes. Additionally, RK inhibited lipid accumulation, and adipogenic transcription factor and lipogenic protein expressions were all decreased by inhibiting HO-1 or ß-catenin using tin protoporphyrin (SnPP) or ß-catenin short-interfering RNA (siRNA), respectively. Furthermore, Wnt10b and ß-catenin expressions were negatively regulation by SnPP. CONCLUSION: RK may exert anti-adipogenic effects through modulation of the HO-1/Wnt/beta-catenin signaling pathway.