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Cytotoxic T lymphocytes (CTLs) mediate host defense against viral and intracellular bacterial infections and tumors. However, the magnitude of CTL response and their function needed to confer heterosubtypic immunity against influenza virus infection are unknown. We addressed the role of CD8+ T cells in the absence of any cross-reactive antibody responses to influenza viral proteins using an adenoviral vector expressing a 9mer amino acid sequence recognized by CD8+ T cells. Our results indicate that both CD8+ T cell frequency and function are crucial for heterosubtypic immunity. Low morbidity, lower viral lung titers, low to minimal lung pathology, and better survival upon heterosubtypic virus challenge correlated with the increased frequency of NP-specific CTLs. NP-CD8+ T cells induced by differential infection doses displayed distinct RNA transcriptome profiles and functional properties. CD8+ T cells induced by a high dose of influenza virus secreted significantly higher levels of IFN-γ and exhibited higher levels of cytotoxic function. The mice that received NP-CD8+ T cells from the high-dose virus recipients through adoptive transfer had lower viral titers following viral challenge than those induced by the low dose of virus, suggesting differential cellular programming by antigen dose. Enhanced NP-CD8+ T-cell functions induced by a higher dose of influenza virus strongly correlated with the increased expression of cellular and metabolic genes, indicating a shift to a more glycolytic metabolic phenotype. These findings have implications for developing effective T cell vaccines against infectious diseases and cancer. IMPORTANCE: Cytotoxic T lymphocytes (CTLs) are an important component of the adaptive immune system that clears virus-infected cells or tumor cells. Hence, developing next-generation vaccines that induce or recall CTL responses against cancer and infectious diseases is crucial. However, it is not clear if the frequency, function, or both are essential in conferring protection, as in the case of influenza. In this study, we demonstrate that both CTL frequency and function are crucial for providing heterosubtypic immunity to influenza by utilizing an Ad-viral vector expressing a CD8 epitope only to rule out the role of antibodies, single-cell RNA-seq analysis, as well as adoptive transfer experiments. Our findings have implications for developing T cell vaccines against infectious diseases and cancer.
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Antibacterianos , Azitromicina , Farmacorresistencia Bacteriana , Macrólidos , Sífilis , Treponema pallidum , Humanos , Antibacterianos/farmacología , Antibacterianos/provisión & distribución , Antibacterianos/uso terapéutico , Farmacorresistencia Bacteriana/genética , Macrólidos/farmacología , Macrólidos/uso terapéutico , América del Norte , Sífilis/tratamiento farmacológico , Sífilis/genética , Sífilis/microbiología , Treponema pallidum/efectos de los fármacos , Treponema pallidum/genética , Treponema pallidum/aislamiento & purificación , Azitromicina/farmacología , Azitromicina/uso terapéutico , Estados Unidos , Canadá , Penicilina G Benzatina/farmacología , Penicilina G Benzatina/provisión & distribución , Penicilina G Benzatina/uso terapéutico , Doxiciclina/farmacología , Doxiciclina/provisión & distribución , Doxiciclina/uso terapéuticoRESUMEN
BACKGROUND: Sexually transmitted infections (STIs) such as syphilis and HIV remain to be a significant public health issue worldwide. Dual rapid point-of-care tests (POCTs) have shown promise for detecting antibodies to HIV and syphilis but have not been fully evaluated in the field. Our study supported the WHO ProSPeRo study on Sexually Transmitted Infection Point-of-Care Testing (STI POCT) by providing external quality assessment (EQA) for HIV and syphilis testing in reference laboratories and their associated clinical sites in seven countries. METHODS: HIV/syphilis serum liquid and dried tube specimen (DTS) panels were prepared by CDC. Liquid panels were distributed to the reference laboratories for three rounds of testing using commercially and locally available laboratory-based serological tests. DTS panels were sent to the clinical testing sites for 8 rounds of POC testing using the Abbott SD BIOLINE HIV/Syphilis Duo test (hereafter referred to as SD BIOLINE) and the Chembio Dual Path Platform (DPP) HIV-Syphilis assay. EQA panels were tested at CDC using the Rapid Plasma Reagin (RPR) test and the Treponema pallidum Particle Agglutination assay (TP-PA) for syphilis antibodies. Genetic Systems HIV-1/HIV-2 Plus O EIA, Geenius HIV Supplemental Assay and the Oraquick Advance HIV test were used to detect HIV antibodies in the EQA panels. Results from the reference laboratories and POCT sites were compared to those obtained at the CDC and a percentage agreement was calculated. RESULTS: Qualitative RPR and TP-PA performed at the reference laboratories demonstrated 95.4-100% agreement with CDC results while quantitative RPR and TP-PA tests demonstrated 87.7% and 89.2% agreement, respectively. A 93.8% concordance rate was observed for qualitative HIV testing in laboratories. EQA testing at clinical sites using dual tests showed 98.7% and 99.1% agreement for detection of HIV antibodies and eight out of 10 sites had > 95.8% agreement for syphilis testing. However, two clinical sites showed only 65.0-66.7% agreement for SD BIOLINE and 84.0-86.7% for DPP, respectively, for syphilis testing. CONCLUSIONS: Overall, laboratories demonstrated high EQA performance in this study. Both HIV/syphilis POCTs gave expected results in the clinic-based evaluations using DTS. However, testing errors were identified in a few testing sites suggesting the necessity for continuous training and monitoring the quality of POC testing.
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Infecciones por VIH , VIH-1 , Sífilis , Humanos , Treponema pallidum , Anticuerpos Anti-VIH , Infecciones por VIH/diagnóstico , Sensibilidad y Especificidad , Anticuerpos Antibacterianos , Pruebas en el Punto de Atención , Serodiagnóstico de la Sífilis/métodos , VIH-2 , Organización Mundial de la Salud , Sistemas de Atención de PuntoRESUMEN
Estrogen-induced intrahepatic cholestasis (IHC) is a mild but potentially serious risk and urges for new therapeutic targets and effective treatment. Our previous study demonstrated that RORγt and CXCR3 signaling pathway of invariant natural killer T (iNKT) 17 cells play pathogenic roles in 17α-ethinylestradiol (EE)-induced IHC. Ursodeoxycholic acid (UDCA) and 18ß-glycyrrhetinic acid (GA) present a protective effect on IHC partially due to their immunomodulatory properties. Hence in present study, we aim to investigate the effectiveness of UDCA and 18ß-GA in vitro and verify the accessibility of the above targets. Biochemical index measurement indicated that UDCA and 18ß-GA presented efficacy to alleviate EE-induced cholestatic cytotoxicity. Both UDCA and 18ß-GA exhibited suppression on the CXCL9/10-CXCR3 axis, and significantly restrained the expression of RORγt in vitro. In conclusion, our observations provide new therapeutic targets of UDCA and 18ß-GA, and 18ß-GA as an alternative treatment for EE-induced cholestasis.
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Colestasis , Ácido Glicirretínico , Células T Asesinas Naturales , Receptores CXCR3 , Ácido Ursodesoxicólico , Colestasis/inducido químicamente , Colestasis/tratamiento farmacológico , Etinilestradiol/toxicidad , Ácido Glicirretínico/análogos & derivados , Ácido Glicirretínico/farmacología , Ácido Glicirretínico/uso terapéutico , Miembro 3 del Grupo F de la Subfamilia 1 de Receptores Nucleares/genética , Receptores CXCR3/genética , Receptores CXCR3/metabolismo , Transducción de Señal , Ácido Ursodesoxicólico/farmacología , Ácido Ursodesoxicólico/uso terapéutico , Animales , RatonesRESUMEN
INTRODUCTION: The reemergence of syphilis, especially congenital syphilis, presents a significant public health threat. Accurate diagnosis of syphilis depends on recognition of a constellation of symptoms, review of medical and sexual history, and multiple laboratory tests. While reliable, current tests for syphilis can be difficult to interpret, which can lead to delays in treatment. AREA COVERED: This review summarizes the major advantages and limitations of available diagnostic laboratory methods for syphilis, provides an update on recent advances in laboratory tools, and highlights the urgent need for coordinated efforts to create new tools to halt the resurgence of syphilis. EXPERT OPINION: In syphilis, the wide variety of short-lived signs and symptoms followed by periods of latency create diagnostic challenges. Currently available laboratory tests, when positive, require additional information to interpret (prior testing, treatment, and sexual history). Point-of-care tests that can rapidly and accurately detect both treponemal and non-treponemal antibodies would be a huge step toward reducing test turnaround time and time to treatment. Incorporating biological insights and technology innovations to advance the development of direct detection assays is urgently needed. A comprehensive coordinated effort is critical to stem the tide of rising syphilis in the United States and globally.
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Sífilis Congénita , Sífilis , Humanos , Sífilis/diagnóstico , Sífilis Congénita/diagnóstico , Treponema pallidum , Serodiagnóstico de la Sífilis/métodos , Sensibilidad y Especificidad , Anticuerpos AntibacterianosRESUMEN
Hyperlipidemia acute pancreatitis (HLAP) is a specific type of pancreatitis mainly caused by elevated serum triglyceride (TG) levels. Therefore, knowledge of patients' medical history is crucial to the identification of those at high risk of HLAP. Diabetes and obesity are associated with high levels of triglycerides, a risk factor for the development of HLAP, which should be controlled before pregnancy. Moreover, HLAP is associated with additional diagnostic and management challenges related to hyperlipidemia (HL) and pregnancy. HLAP during pregnancy has a rapid onset and rapid progression, and complications are more likely to damage the function of multiple organs. HLAP is more common after 28 weeks of pregnancy, the cause is mostly high TG and the serum TG of the patient is often >1,000 mg/d1. Clinicians should be alert to the occurrence of server acute pancreatitis (AP). Therefore, clinicians need to identify and implement effective treatment in a timely manner to control the progression of HLAP during pregnancy and improve pregnancy outcomes. The present study reported the case of a 26-year-old pregnant patient who was hospitalized for epigastric pain at 35 weeks and 2 days of gestation. Medical and family history reported previous diagnoses of diabetes and obesity (weight before pregnancy, 103 kg; BMI, 36.40 kg/m2). Laboratory tests demonstrated high levels of lipase and amylase, a notable systemic inflammatory response, HL, coagulopathy, hypoproteinemia and hyperglycemia. Abdominal ultrasonography demonstrated a hypoechoic pancreatic head. A clinical diagnosis of AP was confirmed using CT scanning. Initial interventions for HLAP included aggressive intravenous hydration, bowel rest, pain control and a combination of heparin and insulin. Lipid-lowering agents were administered to reduce serum lipid levels. Hemoperfusion and continuous renal replacement therapy were also used to rapidly counteract the elevated lipid levels. Antibiotics were administered in the present case because inflammatory markers such as leukocytes, neutrophils and C-reactive protein were elevated. The patient and newborn were discharged 11 days after hospitalization, with an improvement in maternal clinical health and the infant was healthy. When evaluating pregnant patients with pre-obesity and diabetes presenting with abdominal pain, obstetricians should consider HLAP. Timely diagnosis and multi-team precision treatment are effective for good outcomes for mother and baby.
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The Internet of Things (IoT) is an advanced technology that comprises numerous devices with carrying sensors to collect, send, and receive data. Due to its vast popularity and efficiency, it is employed in collecting crucial data for the health sector. As the sensors generate huge amounts of data, it is better for the data to be aggregated before being transmitting the data further. These sensors generate redundant data frequently and transmit the same values again and again unless there is no variation in the data. The base scheme has no mechanism to comprehend duplicate data. This problem has a negative effect on the performance of heterogeneous networks.It increases energy consumption; and requires high control overhead, and additional transmission slots are required to send data. To address the above-mentioned challenges posed by duplicate data in the IoT-based health sector, this paper presents a fuzzy data aggregation system (FDAS) that aggregates data proficiently and reduces the same range of normal data sizes to increase network performance and decrease energy consumption. The appropriate parent node is selected by implementing fuzzy logic, considering important input parameters that are crucial from the parent node selection perspective and share Boolean digit 0 for the redundant values to store in a repository for future use. This increases the network lifespan by reducing the energy consumption of sensors in heterogeneous environments. Therefore, when the complexity of the environment surges, the efficiency of FDAS remains stable. The performance of the proposed scheme has been validated using the network simulator and compared with base schemes. According to the findings, the proposed technique (FDAS) dominates in terms of reducing energy consumption in both phases, achieves better aggregation, reduces control overhead, and requires the fewest transmission slots.
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In this paper, we proposed a multibeam switching antenna based on a low-cost reconfigurable hybrid metasurface applied for beamforming systems. The antenna consists of two parts: a microstrip feed antenna and a transmission hybrid metasurface. The latter is composed of three types of units with different amplitude and phase responses to electromagnetic waves so as to control the beams of the feed antenna. Sixteen PIN diodes are arranged in the metasurface with a simple bias network. When two different direct-current voltages are applied to the PIN diodes, the antenna can dynamically switch between two beams and four beams. For demonstration, the proposed antenna is fabricated, and the measured results show that the antenna operates at 9.07-9.42 GHz (-10 dB bandwidth) with a total size of 1.80λ0 × 1.52λ0 × 0.22λ0 (λ0 corresponds to the wavelength of 9.28 GHz in free space). With the merits of a compact structure, low cost and good radiation performance, the proposed design is suitable for beamforming applications.
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Due to the growing scarcity of spectrum resources in the low-frequency band, the requirement of beam-reconfigurable antennas in the millimeter wave band is urgent. In this paper, a W-band graphene-based metasurface working in a broad bandwidth is proposed with reflective amplitude coding. Here, graphene sheets play a dual role in radiating and regulating electromagnetic waves. By adjusting the Fermi levels of graphene, the reflective amplitude and phase of the metasurface can be modulated simultaneously, enabling multi-beam switching and beam deflection in far-field. The proposed metasurface achieves amplitude-phase modulation within a significantly wide bandwidth which covers 75-91.5 GHz and 99.3-115 GHz. By optimizing the coding patterns, the proposed graphene-based metasurfaces are able to not only realize 2-D beam steering, but also achieve beam switching from single beam to four beams at 87 GHz. The proposed design provides a novel solution for the flexible manipulation of millimeter waves, which can be applied to various fields such as vehicle radar, satellite communication, 6G wireless communication, and beyond.
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Background: Emerging data suggest that second-generation influenza vaccines with higher hemagglutinin (HA) antigen content and/or different production methods may induce stronger antibody responses to HA than standard-dose egg-based influenza vaccines in adults. We compared antibody responses to high-dose egg-based inactivated (HD-IIV3), recombinant (RIV4), and cell culture-based (ccIIV4) vs standard-dose egg-based inactivated influenza vaccine (SD-IIV4) among health care personnel (HCP) aged 18-65 years in 2 influenza seasons (2018-2019, 2019-2020). Methods: In the second trial season, newly and re-enrolled HCPs who received SD-IIV4 in season 1 were randomized to receive RIV4, ccIIV4, or SD-IIV4 or were enrolled in an off-label, nonrandomized arm to receive HD-IIV3. Prevaccination and 1-month-postvaccination sera were tested by hemagglutination inhibition (HI) assay against 4 cell culture propagated vaccine reference viruses. Primary outcomes, adjusted for study site and baseline HI titer, were seroconversion rate (SCR), geometric mean titers (GMTs), mean fold rise (MFR), and GMT ratios that compared vaccine groups to SD-IIV4. Results: Among 390 HCP in the per-protocol population, 79 received HD-IIV3, 103 RIV4, 106 ccIIV4, and 102 SD-IIV4. HD-IIV3 recipients had similar postvaccination antibody titers compared with SD-IIV4 recipients, whereas RIV4 recipients had significantly higher 1-month-postvaccination antibody titers against vaccine reference viruses for all outcomes. Conclusions: HD-IIV3 did not induce higher antibody responses than SD-IIV4, but, consistent with previous studies, RIV4 was associated with higher postvaccination antibody titers. These findings suggest that recombinant vaccines rather than vaccines with higher egg-based antigen doses may provide improved antibody responses in highly vaccinated populations.
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In recent years, many intriguing electromagnetic (EM) phenomena have come into being utilizing metasurfaces (MSs). However, most of them operate in either transmission or reflection mode, leaving the other half of the EM space completely unmodulated. Here, a kind of transmission-reflection-integrated multifunctional passive MS is proposed for entire-space electromagnetic wave manipulation, which can transmit the x-polarized EM wave and reflect the y-polarized EM wave from the upper and lower space, respectively. By introducing an H-shaped chiral grating-like micro-structure and open square patches into the unit, the MS acts not only as an efficient converter of linear-to-left-hand circular (LP-to-LHCP), linear-to-orthogonal (LP-to-XP), and linear-to-right-hand circular (LP-to-RHCP) polarization within the frequency bands of 3.05-3.25, 3.45-3.8, and 6.45-6.85 GHz, respectively, under the x-polarized EM wave, but also as an artificial magnetic conductor (AMC) within the frequency band of 12.6-13.5 GHz under the y-polarized EM wave. Additionally, the LP-to-XP polarization conversion ratio (PCR) is up to -0.52 dB at 3.8 GHz. To discuss the multiple functions of the elements to manipulate EM waves, the MS operating in transmission and reflection modes is designed and simulated. Furthermore, the proposed multifunctional passive MS is fabricated and experimentally measured. Both measured and simulated results confirm the prominent properties of the proposed MS, which validates the design's viability. This design offers an efficient way to achieve multifunctional meta-devices, which may have latent applications in modern integrated systems.
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Automated nontreponemal rapid plasma reagin (RPR) tests were recently introduced in the United States for syphilis testing and limited performance data are available. In collaboration with the Association of Public Health Laboratories, three public health laboratories (PHL) were chosen through a competitive selection process to evaluate the performance of three FDA-cleared automated RPR test systems: BioPlex 2200 Syphilis Total & RPR assay (Bio-Rad Laboratories), AIX 1000 (Gold Standard Diagnostics), and ASI Evolution (Arlington Scientific). Panels prepared at the CDC included: a qualitative panel comprised of 734 syphilis reactive/nonreactive sera; a quantitative panel of 50 syphilis reactive sera (RPR titer 1:64 to 1:1,024); and a reproducibility panel of 15 nonreactive and reactive sera (RPR titer 1:1 to 1:64). Panels were shipped frozen to the PHL and tested on the automated RPR systems following manufacturers' instructions. Prior test results were blinded to all laboratories. When compared to manual RPR (Arlington Scientific) performed at the CDC as a reference test, the qualitative panel results demonstrated an overall concordance of 95.9% for AIX 1000, 94.6% for ASI Evolution, and 92.6% for Bioplex RPR; quantitative panel showed within range titer of 2-fold for 94% of specimens for AIX 1000, 68% for ASI Evolution, and 64% for BioPlex RPR, and the reproducibility testing panel demonstrated point estimates ranging from 69 to 95%. Automated RPR instruments could reduce turnaround time and minimize interpretation errors. However, additional evaluations with more specimens could assist laboratories with implementing automated RPR tests and understanding their limitations.
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Sífilis , Humanos , Sífilis/diagnóstico , Reaginas , Reproducibilidad de los Resultados , Anticuerpos Antibacterianos , Serodiagnóstico de la Sífilis/métodos , Treponema pallidumRESUMEN
BACKGROUND: Antibody responses to non-egg-based standard-dose cell-culture influenza vaccine (containing 15 µg hemagglutinin [HA]/component) and recombinant vaccine (containing 45 µg HA/component) during consecutive seasons have not been studied in the United States. METHODS: In a randomized trial of immunogenicity of quadrivalent influenza vaccines among healthcare personnel (HCP) aged 18-64 years over 2 consecutive seasons, HCP who received recombinant-HA influenza vaccine (RIV) or cell culture-based inactivated influenza vaccine (ccIIV) during the first season (year 1) were re-randomized the second season of 2019-2020 (year 2 [Y2]) to receive ccIIV or RIV, resulting in 4 ccIIV/RIV combinations. In Y2, hemagglutination inhibition antibody titers against reference cell-grown vaccine viruses were compared in each ccIIV/RIV group with titers among HCP randomized both seasons to receive egg-based, standard-dose inactivated influenza vaccine (IIV) using geometric mean titer (GMT) ratios of Y2 post-vaccination titers. RESULTS: Y2 data from 414 HCP were analyzed per protocol. Compared with 60 IIV/IIV recipients, 74 RIV/RIV and 106 ccIIV/RIV recipients showed significantly elevated GMT ratios (Bonferroni corrected P < .007) against all components except A(H3N2). Post-vaccination GMT ratios for ccIIV/ccIIV and RIV/ccIIV were not significantly elevated compared with IIV/IIV except for RIV/ccIIV against A(H1N1)pdm09. CONCLUSIONS: In adult HCP, receipt of RIV in 2 consecutive seasons or the second season was more immunogenic than consecutive egg-based IIV for 3 of the 4 components of quadrivalent vaccine. Immunogenicity of ccIIV/ccIIV was similar to that of IIV/IIV. Differences in HA antigen content may play a role in immunogenicity of influenza vaccination in consecutive seasons. CLINICAL TRIALS REGISTRATION: NCT03722589.
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Subtipo H1N1 del Virus de la Influenza A , Vacunas contra la Influenza , Gripe Humana , Vacuna contra Viruela , Adulto , Humanos , Anticuerpos Antivirales , Técnicas de Cultivo de Célula , Atención a la Salud , Pruebas de Inhibición de Hemaglutinación , Subtipo H3N2 del Virus de la Influenza A , Estados Unidos , Vacunación , Vacunas Combinadas , Vacunas de Productos Inactivados , Vacunas SintéticasRESUMEN
In this study we demonstrate a high-performance polarization rotator (PR) based on flat-shaped photonic crystal fiber. The flat surfaces of the fiber are plated on gold films as electrodes, and the core of the structure is filled with liquid crystal. The polarization rotation characteristics of the flat-shaped fiber can be effectively adjusted by applying external voltage. The optical properties are analyzed using the finite element method (FEM). The results show that the magnitude of the modulation voltage is closely related to the thickness of the flat fiber. When the fiber thickness is 20 µm, only 100 V is required to achieve the highest PR performance. In the wavelength of the 1.55 µm band (~200 nm bandwidth), the conversion length of the PR is only 3.99 µm, the conversion efficiency is close to 100%, and the minimum crosstalk value is -26.2 dB. The presented PR, with its excellent performance, might enable promising applications in the communication system and the photonic integrated circuits.
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Beam splitters play important roles in several optical systems. Due to the growing demand for the miniaturization of optical systems, it is necessary to design beam splitters with nanoscale dimensions to miniaturize the essential components for integrated optical circuits. In this work, we propose and numerically demonstrate a broadband, high efficient, and four-channel beam splitter based on a fishnet-shaped metasurface. The proposed structure is constructed of cruciform AlSb nanoantennas on the PDMS substrate. The simple design can split a beam of light into four beams with equal intensity, it achieves a conversion efficiency above 83%, and an anomalous transmission intensity exceeding 0.8 for the wavelength range of 761-835 nm. In this wavelength range, the beam splitting angle changes from 46.45° to 53.68°. Moreover, the four-channel beam splitter is tunable when the metasurface is designed as a discrete structure. At the wavelength of 874 nm, the beam splitting angle can be adjusted from 56.34° to 46.39° as the period increases from 1050 nm to 1207 nm by stretching the substrate. The presented metasurface might enable promising applications in integrated optical devices, owing to its advantages of multi-channel, wide broadband, high efficiency, and large beam split angle.
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Beam splitters are vital components in several optical systems. It is highly desirable, and compact beam splitters with ultra-broadband performances, high efficiencies, and large split angles are still being sought. In this paper, we demonstrate and numerically investigate an ultra-broadband and highly efficient optical beam splitter based on a quasi-continuous metasurface. The proposed design is constructed of quasi-continuous triangle-shaped gallium phosphide nanoantennas on a silica substrate. The simple structure can achieve a conversion efficiency and an anomalous transmission intensity above 90% and 0.8 covering the wavelength range of 1537-1826 nm, respectively. The maximum beam split angle in the operating bandwidth reaches 131.84° at the wavelength of 1826 nm. Particularly, the operating bandwidth is still as high as 125 nm with the anomalous transmission intensity above 0.92 and the conversion efficiency exceeding 99%. Moreover, the results show that the performance of the metasurface-based optical beam splitter can be further enhanced by optimizing structural parameters. We also demonstrate the adjustability of the beam splitter by adding refractive index (RI) materials on the surface of the device. The results show that the incident plane wave can be divided into three beams with intensity adjustability. The presented metasurface is very promising in the fields of multiplexers, interferometers, and optical communications, owing to its advantages of ultra-broadband, highly efficient, and large split angle simultaneously.
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Background: The incidence of esophageal squamous cell carcinoma in China ranks first in the world. The early diagnosis technology is underdeveloped, and the prognosis is poor, which seriously threatens the quality of life of the Chinese people. Epidemiological findings are related to factors such as diet, living habits, and age. The specific mechanism is not clear yet. Metabolomics is a kind of omics that simultaneously and quantitatively analyzes the comprehensive profile of metabolites in living systems. It has unique advantages in the study of the diagnosis and pathogenesis of tumor-related diseases, especially in the search for biomarkers. Therefore, it is desirable to perform metabolic profiling analysis of cancer tissues through metabolomics to find potential biomarkers for the diagnosis and treatment of esophageal squamous cell carcinoma. Methods: HPLC-TOF-MS/MS technology and Illumina Hiseq Xten Sequencing was used for the analysis of 210 pairs of matched esophageal squamous cell carcinoma tissues and normal tissues in Zhenjiang City, Jiangsu Province, a high-incidence area of esophageal cancer in China. Bioinformatics analysis was also performed. Results: Through metabolomic and transcriptomic analysis, this study found that a total of 269 differential metabolites were obtained in esophageal squamous cell carcinoma and normal tissues, and 48 differential metabolic pathways were obtained through KEGG enrichment analysis. After further screening and identification, 12 metabolites with potential biomarkers to differentiate esophageal squamous cell carcinoma from normal tissues were obtained. Conclusions: From the metabolomic data, 4 unknown compounds were found to be abnormally expressed in esophageal squamous cell carcinoma for the first time, such as 9,10-epoxy-12,15-octadecadienoate; 3 metabolites were found in multiple abnormal expression in another tumor, but upregulation or downregulation was found for the first time in esophageal cancer, such as oleoyl glycine; at the same time, it was further confirmed that five metabolites were abnormally expressed in esophageal squamous cell carcinoma, which was similar to the results of other studies, such as PE.
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Carcinoma de Células Escamosas , Neoplasias Esofágicas , Carcinoma de Células Escamosas de Esófago , Biomarcadores de Tumor/genética , Carcinoma de Células Escamosas/diagnóstico , Carcinoma de Células Escamosas/epidemiología , Carcinoma de Células Escamosas/genética , Neoplasias Esofágicas/diagnóstico , Neoplasias Esofágicas/epidemiología , Neoplasias Esofágicas/genética , Carcinoma de Células Escamosas de Esófago/diagnóstico , Carcinoma de Células Escamosas de Esófago/genética , Humanos , Calidad de Vida , Espectrometría de Masas en TándemRESUMEN
Downstream next-generation sequencing (NGS) of the syphilis spirochete Treponema pallidum subspecies pallidum (T. pallidum) is hindered by low bacterial loads and the overwhelming presence of background metagenomic DNA in clinical specimens. In this study, we investigated selective whole-genome amplification (SWGA) utilizing multiple displacement amplification (MDA) in conjunction with custom oligonucleotides with an increased specificity for the T. pallidum genome and the capture and removal of 5'-C-phosphate-G-3' (CpG) methylated host DNA using the NEBNext Microbiome DNA enrichment kit followed by MDA with the REPLI-g single cell kit as enrichment methods to improve the yields of T. pallidum DNA in isolates and lesion specimens from syphilis patients. Sequencing was performed using the Illumina MiSeq v2 500 cycle or NovaSeq 6000 SP platform. These two enrichment methods led to 93 to 98% genome coverage at 5 reads/site in 5 clinical specimens from the United States and rabbit-propagated isolates, containing >14 T. pallidum genomic copies/µL of sample for SWGA and >129 genomic copies/µL for CpG methylation capture with MDA. Variant analysis using sequencing data derived from SWGA-enriched specimens showed that all 5 clinical strains had the A2058G mutation associated with azithromycin resistance. SWGA is a robust method that allows direct whole-genome sequencing (WGS) of specimens containing very low numbers of T. pallidum, which has been challenging until now. IMPORTANCE Syphilis is a sexually transmitted, disseminated acute and chronic infection caused by the bacterial pathogen Treponema pallidum subspecies pallidum. Primary syphilis typically presents as single or multiple mucocutaneous lesions and, if left untreated, can progress through multiple stages with various clinical manifestations. Molecular studies often rely on direct amplification of DNA sequences from clinical specimens; however, this can be impacted by inadequate samples due to disease progression or timing of patients seeking clinical care. While genotyping has provided important data on circulating strains over the past 2 decades, WGS data are needed to better understand strain diversity, perform evolutionary tracing, and monitor antimicrobial resistance markers. The significance of our research is the development of an SWGA DNA enrichment method that expands the range of clinical specimens that can be directly sequenced to include samples with low numbers of T. pallidum.