Mecanismo y diseño de vacunas para el sars-cov-2, revisión narrativa / Mechanism and design of vaccines against sars-cov-2, narrative review / Mecanismo e desenho de vacinas para sars-cov-2, revisão narrativa

La infección por el virus SARS-CoV-2, conocida como COVID-19, ha causado alta morbilidad y mortalidad en el mundo. Después de haber descifrado el código genético del virus y haber desarrollado un gran trabajo investigativo en la creación de vacunas, con diversas estrategias de acción, se ha logrado disminuir la morbi mortalidad. Fue necesario acelerar el proceso de producción de vacunas, lo cual estuvo facilitado por el avanzado conocimiento científico en el campo de la genética y la virología, para brindar a la especie humana una protección eficaz y segura contra la agresiva y progresiva infección. Las vacunas se clasifican de acuerdo con su mecanismo de acción, existen vacunas basadas en vectores virales que no se replican, vacunas recombinantes, otras basadas en virus atenuados y virus inactivos, y (la gran novedad de la ciencia actual) las vacunas basadas en ARN mensajero y ADN. Estas últimas han demostrado una gran eficacia y seguridad en la prevención de la infección por el SARS-CoV-2, también han impactado de manera fuerte, por lo que han reducido la infección y la mortalidad en la población. En consecuencia, cada día que pasa desde que se inició el periodo de vacunación mundial, se evidencia una reducción en la curva de contagio y mortalidad por COVID-19.

The infection produced by the SARS-CoV-2 virus, known as COVID-19, has caused high morbidity and mortality across the world. After having deciphered the virus's genoma and carried out investigative endeavors that led to the creation of a variety of vaccines with different mechanisms of action, it has been possible to decrease the morbidity and mortality associated with the virus. It was necessary to accelerate the vaccine production process, which was facilitated by advanced scientific knowledge within the disciplines of genetics and virology, in order to provide the human species with a safe and effective form of protection against the aggressive and progressive infection. Vaccines are classified differently depending on their action mechanisms: there are some based on non-replicating viral vectors, recombinant vaccines, ones that are based on attenuated or inactivated viruses, and (the greatest novelty of current scientific developments) vaccines based on DNA and messenger RNA. The latter has demonstrated significant efficacy and safety in the prevention of the SARS-CoV-2 infection as observed in preliminary studies, and they have meaningfully impacted the population by reducing the rates of infection and mortality. As a result, decreased levels of spread of and mortality from COVID-19 have been evidenced across the globe following the beginning of the vaccine distribution period.

A infecção pelo vírus SARS-CoV-2, conhecido como COVID-19, tem causado elevada morbidade e mortalidade no mundo. Depois de ter decifrado o código genético do virus e de ter realizado um grande trabalho de investigação na criação de vacinas, com diversas estratégias de ação, a morbilidade e a mortalidade foram reduzidas. Foi necessário acelerar o processo de produção de vacinas, facilitado por conhecimentos científicos avançados no domínio da genética e da virologia, para proporcionar à espécie humana uma proteção eficaz e segura contra a infecção agressiva e progressiva. As vacinas são classificadas de acordo com seu mecanismo de ação, existem vacinas baseadas em vetores virais que não se replicam, vacinas recombinantes, outras baseadas em virus atenuados e vírus inativos, e (a grande novidade da ciência atual) vacinas baseadas em RNA mensageiro e ADN. Estas últimas demonstraram grande eficácia e segurança na prevenção da infecção por SARS-CoV-2, mas também tiveram um forte impacto, razão pela qual reduziram a infecção e a mortalidade na população. Consequentemente, a cada dia que passa desde o início do período global de vacinação, fica evidente uma redução na curva de contágio e mortalidade por COVID-19.

Discussion on results of national measurement capability comparison assessment for gross α and gross β radioactivity in water and calculation formulas for spike recovery / 中国辐射卫生

Objective To summarize and analyze the difficulties and key points in the measurement of gross α and gross β radioactivity in water based on the results of national measurement capability comparison assessment, and provide the basis and reference for the future work and the development of new local standards. Methods The research team participated in the comparison assessment for measurement of the gross radioactivity in water samples organized by National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention. According to the comparison assessment results and the content in the national standard GB/T 5750.13—2023 (published draft), the steps of spike recovery involved during the measurement were analyzed and discussed. Two different formulas used for spike recovery calculation were analyzed for their impact on the final measurement results. Results When the spike recovery F(derived) derived from the formulas was used for result calculation, the spike recovery ranged as follows: gross α: 63.00%−84.60%, and gross β: 95.0%−99.1%; 3/6 of the comparison results were determined as excellent and 3/6 as pass as a whole (among them, 4 were excellent and 2 were pass for both single gross α assessment items and single gross β assessment items). When the spike recovery F from the GB/T 5750.13—2023 (published draft) was used for result calculation, the spike recovery ranged as follows: gross α: 39.69%−71.57%, and gross β: 90.25%−98.21%; 5/6 of the comparison results were determined as fail and 1/6 as pass (among them, 5 were fail and 1 was pass for single gross α assessment items; 5 were excellent and 1 was pass for single gross β assessment items). When two different formulas were used for spike recovery calculation, there was a significant difference in gross α radioactivity measurement (t = 4.27, P = 0.03 < 0.05), but there was no significant difference in gross β radioactivity measurement (t = 0.667, P = 0.524 > 0.05). Conclusion In the measurement of gross α and gross β radioactivity in water, appropriate reference to the spike recovery has a great influence on the measurement results. Therefore, quality control should be strengthened to further ensure the accuracy of measurement.

Recent advances in drug screening methods of SARS-CoV-2 spike protein / 药学学报

The pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a serious impact on global public health and the economy. SARS-CoV-2 infiltrates host cells via its surface spike protein, which binds to angiotensin-converting enzyme 2 on the host cell membrane. As a result, small molecules targeting spike protein have emerged as a hotspot in anti-SARS-CoV-2 drug research. Activity screening is an important step in seeking small molecule drugs. Therefore, this article aims to review the biological activity evaluation methods of small molecule inhibitors targeting SARS-CoV-2 spike protein, with the goal of laying the foundation for the discovery of new anti-SARS-CoV-2 drugs.

Identificación de la proteína "Spike" en sangre de cordón umbilical de recién nacidos de madres vacunadas contra SARS-CoV-2 durante el embarazo / Identification of the "Spike" protein in umbilical cord blood of newborns from mothers vaccinated against SARS-CoV-2 during pregnancy

Resumen OBJETIVO: Identificar la coexistencia de anticuerpos contra la proteína "Spike" del SARS-CoV-2 en sangre del cordón umbilical de recién nacidos de madres vacunadas contra la COVID-19 durante el embarazo. MATERIALES Y MÉTODOS: Estudio transversal, llevado a cabo en un centro de atención privada de la Ciudad de México, entre junio del 2021 y febrero del 2022, al que se incluyeron mujeres que completaron el esquema de vacunación contra SARS-CoV-2 durante el embarazo y en quienes en el trascurso de la finalización del embarazo parto o cesárea se haya obtenido una muestra de sangre de cordón umbilical para identificar la proteína Spike. RESULTADOS: Se registraron 219 pacientes y se analizaron 5 tipos de vacuna de dos plataformas diferentes; Pfizer®, Moderna®, AstraZeneca®, J&J/Janssen® y Sputnik®. La concentración de anticuerpos fue mayor en pacientes vacunadas con Moderna® y Sputnik®. Mediante el análisis de regresión de Cox se evaluó un modelo predictivo, sin mostrar diferencia estadísticamente significativa en la cantidad de anticuerpos generados luego de la primera y segunda dosis de la vacuna. CONCLUSIONES: La vacunación prenatal contra la COVID-19 induce una fuerte respuesta humoral materna, que se transfiere de manera efectiva al feto, incluso con una sola dosis.

Abstract OBJECTIVE: Identification of the presence of SARS-CoV-2 Spike protein antibodies in cord blood of newborns following maternal COVID-19 vaccination during pregnancy. MATERIALS AND METHODS: Cross-sectional study in vaccinated women during pregnancy who had a cord blood sample taken during the surgical procedure Vaginal birth/Cesarean section to measure spike protein antibodies. RESULTS: We registered 219 patients in the study: five types of vaccines, from two different platforms, were tested: Pfizer®, Moderna®, AstraZeneca®, J&J/Janssen® and Sputnik®. The levels of antibodies were greater in those patients vaccinated with Moderna® and Sputnik®. Through a Cox regression we made a predictive model where we observed that there are no differences in the number of antibodies generated after the first and second vaccine dose. CONCLUSIONS: Prenatal vaccination against COVID-19 induces a strong maternal humoral response that is effectively transferred to the fetus, even with a single dose.

Varietal Effect of Different Nitrogen Levels on Yield Attributing Characters and Quality Parameters of Wheat (Triticum aestivum L.) Crop

The field experiment was conducted at Agronomy Research FaTArm, Narendra Deva University of Agriculture and Technology, Narendra Nagar (Kumarganj) Faizabad (U.P.), during Rabi season of 2014-2015. The yield components like number of spikes m-2, spike length (cm), number of grain spike-1, grain weight spike-1(g), grain yield (kg ha-1), straw yield (q ha-1) and nitrogen contain in grain (%), nitrogen contain in straw (%), nitrogen uptake in grain (kg ha-1), nitrogen uptake in straw, protein content in grain (%) were maximum under 160 kg N ha-1 and among the varieties over PBW-373, NW-1014 being at par with HD-2327. The main aim of trial conducted that to know the different varietal parameters of wheat crop.

Los desafíos de la nueva variante ómicron para la salud pública: una revisión de la literatura actual / The challenges of the new omicron variant for public health: a review of the current literature

RESUMEN Introducción: Tras la pandemia del coronavirus del síndrome respiratorio agudo grave tipo 2 (SARS-Cov-2), múltiples variantes dentro de la estructura molecular del virus se han venido presentando, las cuales pueden condicionar mayores tasas de contagio, aumento de la morbimortalidad, disminución en la efectividad de las vacunas y manejos farmacológicos. La variante Ómicron, clasificada como variante preocupante (VOC), ha demostrado hasta ahora tener mayor transmisibilidad y una respuesta diferente frente a la vacunación y los esquemas terapéuticos ya instaurados. Objetivo: Describir la variante Ómicron y su impacto sobre la transmisibilidad y mortalidad. Métodos: Revisión de la literatura, en las bases de datos PubMed y Scopus, incluyendo estudios empíricos en inglés y español. De la bibliografía obtenida se utilizaron y analizaron detalladamente 49 artículos. Resultados: Los resultados obtenidos nos permiten identificar los diferentes aspectos de la variante Ómicron, desde su estructura molecular, transmisibilidad hasta su manejo actual y la efectividad de las vacunas contra la misma. Conclusiones: En la actualidad, la variante Ómicron ha constituido un desafío para el sistema de salud por su alta transmisibilidad. Sin embargo, los esfuerzos terapéuticos han demostrado un impacto en severidad y curso de la enfermedad.

ABSTRACT Introduction: Since the pandemic of severe acute respiratory syndrome type 2 coronavi-rus (SARS-Cov-2), multiple variants within the molecular structure of the virus have been appearing, which may lead to higher infection rates, increased morbidity, and mortality, decreased effectiveness of vaccines and pharmacological management. The Omicron variant, classified as variant of concern (VOC) has so far shown higher transmissibility and a different response to vaccination and therapeutic regimens. Objective: To describe the Omicron variant and its impact on transmissibility and mortality. Methods: Literature review in PubMed and Scopus databases, including empirical studies in English and Spanish. From the bibliography obtained, 49 articles were used and analyzed in detail. Results: The results obtained allow us to identify the different aspects of the Omicron variant, from its molecular structure, transmissibility to its current management and the effectiveness of vaccines against it. Conclusions: Currently, the Omicron variant has constituted a challenge for the health system due to its high transmissibility. However, therapeutic efforts have shown an impact on the severity and course of the disease.

Performance of Gladiolus Genotypes for Growth and Flowering under Gangetic Region of Bihar, India

An evaluation experiment on performance of gladiolus genotypes was conducted at Botanical Garden, Bihar Agricultural University, Sabour during 2021-2022 for the identification of suitable genotype for the cultivation in Bihar. Twenty genotypes of gladiolus were evaluated for vegetative, floral and vase life parameters under Gangetic region of Bihar conditions. The best genotypes identified in present investigation based on mean performance were White Prosperity (79.73 cm), Red Majesty (79.27 cm), Nova Lux (73.47 cm) and Rose Supreme (73.40 cm) for spike length. Earliness in flowering was observed in Rose Supreme (83.53 days) which was at par with Punjab Dawn (83.60 days). Genotype Kum Kum responded well for number of spikes per plant (2.73), whereas Pusa Shubham for corms production (3.33). Genotype White Prosperity (30.27 days) and Rose Supreme (28.47 days) were recorded long flowering duration.

Varietal Evaluation of Single Petalled Tuberose (Agave amica Medik.) Germplasm for the Growth and Flowering Attributes

The present experiment was to evaluate eight single petaled tuberose germplasm for growth and flowering attributes. All the tuberose germplasm showed a significant variation in the results. In vegetative parameters, the maximum plant height (79.3 cm) was observed in ‘Mexican Single’, while maximum leaf length (35.1 cm) and leaf width (3.4 cm) was observed in ‘Arka Keerthy’ and ‘Bidhan Rajani’, respectively, then high number of leaves seen in the germplasm ‘Mexican Single. In flowering parameters, ‘Nilakottai local” found superior in days taken for spike emergence (64.7 days), spike length (90.9 cm) and rachis length (31.4 cm) respectively. While, floret length (7.28 cm) and Single floret weight (1.57 g) was observed in ‘Bidhan Rajani’.

Encefalopatías epilépticas y del desarrollo. ¿Que hay de nuevo? / Developmental and epileptic encephalopathy. What is new?

Resumen Este artículo no tiene como objetivo el presentar una descripción detallada de cada una de las encefalopatías epilépticas y del desarrollo, sino más bien discutir cam bios recientes en la terminología y criterios diagnósticos de ciertas encefalopatías, en base a una revisión actua lizada de los últimos 10 años. Se analizan cambios importantes en definiciones de síndromes específicos y nuevos tratamientos que han demostrado eficacia en el manejo de crisis convulsivas en estos pacientes. En conclusión: Las nuevas terapias de modulación genética, contribuirán no solo a reducir la carga de crisis epilépticas, sino también a mejorar el pronóstico cognitivo, y por lo tanto la calidad de vida.

Abstract It is not the intend of this article to present a de tailed description of each developmental and epileptic encephalopathy, but to discuss recent changes in the terminology and diagnostic criteria of specific disorders, based on an updated review of the last 10 years. Important changes in the definitions of specific syn dromes and new treatments that have shown efficacy in the management of seizures in these patients are analyzed. In conclusion: New gene modulation therapy will likely improve not only seizure frequency, but also cog nitive outcome and therefore quality of life.

Identification of Lead Molecules against Multi-target of SARS-CoV-2 from Carica papaya L. through In-silico Method

The efficacy of currently used vaccines against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is questionable since the virus is rapidly mutating. A single drug that can simultaneously act on multi-target at different stages of disease causing pathways is the best option to fight against it. The multi-target inhibitory activities of many phytochemicals have been reported. The present investigation was aimed to screen the multi-target inhibitory activity of 215 phytochemicals from Carica papaya L. against three targets of SARS-CoV-2 viz spike protein (SP), main protease (Mpro), RNA-dependent RNA-polymerase (RdRp) and a target from host, angiotensin-converting enzyme 2 (ACE-2) using the docking tool, AutoDock Vina in PyRx 0.8. The docked results with free energy of binding ? -6 kcal/mol were considered active/hit molecules. Of the 215 phytochemicals, 48 have binding energy ? -6 kcal/mol against all the targets. Further molecular interaction between the ligand and targets, pharmacokinetics and ADMET analysis of the top ranked five hits obtained against each target revealed that the compound hesperidin can be selected as the best lead since it has the least binding energy, admissible ADMET and a better binding score than the control drugs. Hesperidin has been used as an approved drug to treat vascular disease. Overall, results revealed that C. papaya is a rich source of phytochemicals with activity on multi-target of SARS-CoV-2 infection and multiplication in the human host.

Coronavirus without Spike Proteins, respectively Mechanism of Christoph's Quantum Nano-scale Irradiation-Emission or Evaporation of Coronavirus Sars-Cov-2-19 by Plasmalyses Coronavirus without Infection Spike Proteins and without Filling Hydrophile Micells of Phospholipidic Membranes of Cellular Wall

Since the Discovery of the structure (space-conformal) significance of ?-helix and the double helix of DNA, in April 1953, in the paper from 25th April 1953 by Francis Crick and James D. Watson, which were published super significant paper focused on this theme, win the Nobel Prize for Medicine and Physiology in 1962. In this 900 words text in the super journal Nature coauthored by Maurice Wilkins, the real discoveries of the structure of DNA were Rosalind Elsie Franklin, so-called DNA´s dark lady, who died only 37 years of Cancer of ovaria. The Really Heroes of Great Scientific Discoveries are namely these, which are in the background, like Grey Eminence, who often gave for Science all, unfortunately, health, freedom, happiness, and their fragile life. The First Analytical method of sequential replication of DNA by method PCR (Polymerase Chain Reaction) was invented by Nobel Prize Winner in 1993 Kary Mullis with M. Smith. The PCR analytical method contributed to the invention of discovery of genetics scissors, and molecular scalpel – the new method of gene editing with a rather unwieldy name CRISPR-Cas 9-based gene editing research at the nanoscale, can do based on DNA sequencing and switch genes off or an insertion. The Nobel Prize in Chemistry in 2020 has finally been awarded to Emmanuelle Charpentier and Jennifer Doudna. Thanks to these genetics-analytical discoveries the whole of Mankind was salvation from Biological Lethal Weapons SARS-Cov2-19 and many mutational virions escaped intentionally or randomized from Biohazard boxes in the biological laboratories or contaminated Nature (Antarctica, Siberia, Aral Sea, Wu-chan) and may be caused by the Warming Global Climate. The same significance as Mullis, Doudna, and Charpentier was the scientific work of Prof. RNDr. Antonín Holy, Dr.Sc., Dr.HC., who discovered virostatics on Cancer of Human papillomavirus (HPV) Vaccine Gardasil 9 and Cervarix, AIDS resp. HIV, Hepatitide and also Coronavirus. The newest message from the current world coronavirus situation is the project of spike protein vaccine Novavax financially and scientifically supported by the Melinda and Bill Gates Foundation.

Optimizing Cow Urine Application Rate at Varying Fertility and Zinc Levels on Wheat (Triticum aestivum L.) Productivity under Dairy Based Farming System in Varanasi Tract

This field investigation was carried out in two consecutive years of 2016-17 and 2017-18 at the Agricultural Research Farm, Banaras Hindu University, Varanasi (U.P.) to assess the effect of cow urine application at varying fertility and zinc levels on growth and yield of wheat (Triticum aestivum L.) under irrigated condition of Varanasi. Results showed that application of the fertilizer at 100% recommended dose of fertilizers (RDF) recorded significantly higher growth parameters viz. plant height, number of tiller m-1 row length; yield attributes viz. number of effective tiller m-2, spike length, grains spike-1 and test weight as well as grain and straw yield than 75% RDF during both the years. Among the zinc levels, zinc applied at 10 kg ha-1 though remained comparable to 5 kg Zn ha-1, recorded significantly higher values of these parameters over control. Cow urine at the rate of 12000 l ha-1 applied equally at sowing, CRI (Crown Root Initiation) and spike emergence (SE) stages gave higher values of plant height, number of tiller m-1 row length, spike length and number of grain spike-1 which being at par with 4000 l ha-1 cow urine each at sowing and CRI, both recorded significantly higher values than control. However, with respect to effective tillers m-2, grain and straw yield, significant increase was noticed with each increment of cow urine, recording maximum at 12000 l ha-1 during both the years.

Screening and identification of single-chain fragment variable against the receptor-binding domain of spike protein of SARS-CoV-2 / 中国生物制品学杂志

@#Objective To construct a single-chain fragment variable(scFv)phage display library against receptor-binding domain(RBD)of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)spike protein(S)to screen specific scFv and identify the function.Methods m RNA was extracted from spleen cells of mice immunized with RBD protein and reversely transcribed into c DNA,with which as template,genes of the hight chain fragment of variable(VH)and light chain fragment of variable(VL)of scFv were amplified and then assembled into scFv gene fragment through splicing overlap extension PCR(SOE-PCR).The scFv gene fragment was inserted to phage vector to construct scFv phage display library.After four rounds of biopanning,the scFv gene with strong binding ability to RBD was screened and expressed recombinantly,purified and identified for biological activity.Results The constructed scFv phage library showed a titer of 6.0×10(11)pfu/m L.After four rounds of biopanning,four scFv strains with strong binding to RBD were selected,namely scFv11,scFv12,scFv25and scFv28.scFv was mainly expressed in the form of inclusion body with a relative molecular mass of about 27 000,a concentration of 2.4 mg/m L and a purity of about 90%,which bound specifically to mouse monoclonal antibody against His labeled by HRP after purification.All four scFv strains bound specifically to RBD recombinant protein,among which the other 3 scFv strains bound to the S protein of wild type and multiple mutant strains except scFv28.All four strains showed dose-dependent interaction with RBD,with affinity dynamic fitting dissociation constants(K_Ds)8.9,5.92,10.67and 2.36 nmol/L,and steady-state fitting dissociation constants(K_Ds)of 5.3,6.5,8.7 and 5.8 nmol/L,respectively.scFv11,scFv12 and scFv25 simultaneously identified three independent RBD polypeptides,including RBD2(S(11)pfu/m L.After four rounds of biopanning,four scFv strains with strong binding to RBD were selected,namely scFv11,scFv12,scFv25and scFv28.scFv was mainly expressed in the form of inclusion body with a relative molecular mass of about 27 000,a concentration of 2.4 mg/m L and a purity of about 90%,which bound specifically to mouse monoclonal antibody against His labeled by HRP after purification.All four scFv strains bound specifically to RBD recombinant protein,among which the other 3 scFv strains bound to the S protein of wild type and multiple mutant strains except scFv28.All four strains showed dose-dependent interaction with RBD,with affinity dynamic fitting dissociation constants(K_Ds)8.9,5.92,10.67and 2.36 nmol/L,and steady-state fitting dissociation constants(K_Ds)of 5.3,6.5,8.7 and 5.8 nmol/L,respectively.scFv11,scFv12 and scFv25 simultaneously identified three independent RBD polypeptides,including RBD2(S(334～353)),RBD9(S(334～353)),RBD9(S(439～458))and RBD13(S(439～458))and RBD13(S(499～518)).Homologous model of scFv constructed by online server SWISS-MODEL showed a good quality and was used for molecular docking.The interface at which scFv11 interacted with RBD only partially coincided with the interaction interface of human angiotensin converting enzyme 2(ACE2)and RBD,and the interaction interfaces of scFv12 and scFv25 with RBD were quite different from that of ACE2.Conclusion In this study,scFv specifically bound to SARS-Co V-2 RBD was screened and prepared through constructing scFv phage library against SARS-CoV-2 RBD,which provided experimental basis for further development of anti-SARS-CoV-2 drugs and detection reagents.

Highly Sensitive Poly-N-isopropylacrylamide Microgel-based Electrochemical Biosensor for the Detection of SARS-COV-2 Spike Protein / 生物医学与环境科学（英文）

OBJECTIVE@#Late 2019 witnessed the outbreak and widespread transmission of coronavirus disease 2019 (COVID-19), a new, highly contagious disease caused by novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Consequently, considerable attention has been paid to the development of new diagnostic tools for the early detection of SARS-CoV-2.@*METHODS@#In this study, a new poly-N-isopropylacrylamide microgel-based electrochemical sensor was explored to detect the SARS-CoV-2 spike protein (S protein) in human saliva. The microgel was composed of a copolymer of N-isopropylacrylamide and acrylic acid, and gold nanoparticles were encapsulated within the microgel through facile and economical fabrication. The electrochemical performance of the sensor was evaluated through differential pulse voltammetry.@*RESULTS@#Under optimal experimental conditions, the linear range of the sensor was 10 -13-10 -9 mg/mL, whereas the detection limit was 9.55 fg/mL. Furthermore, the S protein was instilled in artificial saliva as the infected human saliva model, and the sensing platform showed satisfactory detection capability.@*CONCLUSION@#The sensing platform exhibited excellent specificity and sensitivity in detecting spike protein, indicating its potential application for the time-saving and inexpensive detection of SARS-CoV-2.

Study on the Quality Analysis Method of Schizonepeta Spike Based on Multiple Feature Components"Quality-quantity"Double Standard / 世界科学技术-中医药现代化

Objective To establish a"quality-quantity"double standard control method of Schizonepeta Spike based on the reference material without relying on multiple reference substances.Methods HPLC technology was used to establish the characteristic atlas of Schizonepeta Spike based on the reference traditional Chinese medicinal materials.The chemical constituents of common peaks were identified by Q-TOF-MS technology,and the authenticity of Schizonepeta Spike was determined by the similarity of characteristic peaks of the reference traditional Chinese medicinal materials and the tested traditional Chinese medicinal materials.The relative content of each characteristic peak in different batches of the tested traditional Chinese medicinal materials was calculated by accurate quantition of pulegone.The"Average value-Standard deviation"was taken as the lower limit of the relative content of characteristic peak chemical components relative to pulegone,and the quality of Schizonepeta Spike was determined according to the lower limit of the relative content of characteristic peak.Results The characteristic atlas and content determination method met the requirements of methodology investigation.A total of 11 common chromatographic peaks were determined,and 6 chemical components were identified,which were luteolin,hesperidin,luteolin-7-O-glucuronide,luteolin,geranyl,pulegone.The similarity between the tested traditional Chinese medicinal materials and reference traditional Chinese medicinal materials was greater than 0.90.The lower limit of relative content of chemical components of Schizonepeta Spike characteristic peak was defined.Conclusion The method can identify Schizonepeta Spike clearly and quickly without many reference substances,and provide reference for quality control of Schizonepeta Spike.

Interpretation of concepts related to electrical status epilepticus / 中风与神经疾病杂志

@#Electrical status epilepticus is a special electroencephalogram phenomenon，which means that the spike and slow waves are almost continuously emitted during the wake-sleep phases. Related concepts are epileptic encephalopathy with electrical status epilepticus during slow wave sleep，electrical status epilepticus in sleep，and subclinical electrographic seizures. The above related concepts are widely used in clinical practice，but there is a lack of unified criteria. There are abuses and misuses of these concepts. Clarifying related concepts is of great significance for scientific research and clinical practice.

A study of different calculation methods of spike and wave indexinelectrical status epilepticus during sleep / 中风与神经疾病杂志

@#Objective Spike and wave index (SWI) is of great importance for the diagnosis of electrical status epilepticus during sleep (ESES) in children patients. This study intended to compare four commonly used SWI methods with full night SWI to evaluate their performances. Methods Fifteen patients diagnosed with ESES by electroencephalogram initial report (SWI≥50%) were retrospectively analyzed,and SWI was calculated for the full night sleep(SWI),the first sleep cycle (SWI1),the first 5 minutes of the first sleep cycle (SWI15 min),the first 10 minutes (SWI10 min) and the first 15 minutes (SWI15 min). Wilcox paired test was used to compare SWI with other four SWI methods respectively. Results The SWI of all four methods was significantly different from the full night SWI (p-values of 0.003、0.002、0.002和0.001,respectively,α=0.0125). Among the 15 patients,14 (93.3%) had the trend that SWI decreased with sleep cycles overnight. Conclusion None of the four methods of SWI is a good substitute for full night SWI. The SWI tends to decrease with sleep cycles overnight,and using the first sleep cycle or anepoch of it to calculate SWI is likely to be overestimated.

Characteristics of whole genome analysis of severe acute respiratory syndrome coronavirus 2 during different epidemic periods in Wuxi City / 中华传染病杂志

Objective:To understand the genome sequence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and spike protein variations during different epidemic periods in Wuxi City.Methods:Nucleic acid was extracted from the nasopharyngeal swab samples of six local cases of coronavirus disease 2019 (COVID-19) (from January to February, 2020) and 13 imported cases of COVID-19 (from March to September, 2021) in Wuxi City, and the whole genome was amplified to construct the sequencing library. The second-generation sequencer was used for sequencing. The CLC Genomics Workbench (21 version) software was used to analyze the offline data with NC_045512.2 as the reference strain, and MEGA 7.0 software was used to construct the phylogenetic tree.Identification of type was conducted by Nextstrain typing method and phylogenetic assignment of named global outbreak lineages (Pangolin) typing method.Results:There were five subtypes in Nextstrain and seven subtypes in Pangolin of the nineteen patients with COVID-19. Compared with NC_045512.2, the median nucleotide mutation sites were 29 (range 0 to 42) and amino acid mutation sites were 20 (range 0 to 34). The six local and 13 imported cases had no common nucleotide mutation sites and were in different evolutionary branches. The sequences of the six local cases were highly homologous with the reference strain sequences (NC_045512.2) at the early stage of the pandemic, and the evolutionary distance was close to that of the reference strain. The 13 imported cases were obviously divided into three evolutionary branches (Alpha, Beta, Delta variant).The four Beta variants shared eight amino acid mutation sites in spike protein, and the two Alpha variants shared eight amino acid mutation sites in spike protein, and the seven Delta variants shared five amino acid mutation sites in spike protein.Conclusions:New mutations of SARS-CoV-2 are constantly emerging during the epidemic. The increase of the nucleotide sites number may result in the change of spike protein amino acid. Therefore, the whole-genome sequencing analysis plays an important role in the accurate tracing of epidemic origin and adjustment of prevention and control measures.

Overexpression of SARS-CoV-2 spike protein mediates growth inhibition in human retinal pigment epithelial cells / 中华眼底病杂志

Objective:To observe the inhibition of SARS-CoV-2 spike protein (S-protein) on the proliferation of human retinal pigment epithelium (RPE) cells.Methods:SARS-CoV-2 S-protein gene fragment expression plasmid (p3xflag-S) was constructed and transfected into human RPE, HEK293 cells. DNA sequencing was used for identification, and the expression of Flag-S was detected by Western blot. HEK293 cells were divided into the cells 1, 2, 3 and 4 and transfected with GFP11 plasmid and vector, GFP1-10 plasmid and vector, transfected with GFP11 and pCMV-HA-ACE2 plasmid, GFP1-10 and p3xflag-S plasmid. Cell 1 was co-cultured with cell 2 (control group 1), cell 2 with cell 3 (control group 2), cell 3 with cell 4 (observation group), and cell 1 mixed with cells 2, 3 and 4 (control group 3). Bright-field microscopy and fluorescence microscopy were used to observe cell fusion. RPE cells were divided into control group and overexpression S-protein group. The cell cycle was detected by flow cytometry; the cell proliferation level was detected by Counting Kit 8 (CCK-8); and the S-protein expression level in RPE cells was detected by Western blot. The Student’s t-test was performed for comparison between groups. Results:DNA sequence assay showed that S-protein cDNA was fused with flag-tagged protein. Western blot assay showed that S-protein-related expression was elevated in transfected HEK293 cells compared with untransfected p3xflag-S cells. Large, multinucleated fused cell clusters were visible under bright-field microscopy; multiple nuclear with distinct green fluorescence were visible in the fused cells under fluorescence microscopy. Western blot assay showed elevated S-protein-related expression in transfected p3xflag-S plasmid RPE cells compared to untransfected p3xflag-S plasmid RPE cells. CCK-8 results showed that the proliferative capacity of RPE cells in the S-protein overexpression group was significantly reduced compared with the control group, with statistically significant differences ( t=22.70, 16.75, 23.38; P<0.000 1). The results of flow cytometry showed that the G1 phase cells in the control and overexpression S-protein groups were 41.1 % and 67.0%, respectively; compared with the control group, the G1 phase cells in the overexpression S-protein group were significantly higher, and the difference was statistically significant ( t=4.76, P=0.018). The apoptosis rate was significantly increased in the S-protein overexpression group compared with the control group, and the difference was statistically significant ( t=4.91, P=0.008). Conclusion:Overexpression of the SARS-CoV-2 spike protein reduced the proliferation of human RPE cells.

Research progress of angiotensin converting enzyme 2 co-expression in non-small cell lung cancer and SARS-CoV-2 / 中国胸心血管外科临床杂志

@#Since the first case of corona virus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at the end of 2019, the virus has spread rapidly around the world and has become a global public health problem. In the process of this virus epidemic, compared with the general population, cancer patients are considered to be highly susceptible people, especially the lung cancer patients. Some studies have shown that angiotensin converting enzyme 2 (ACE2) may be the pathway for SARS-CoV-2 to infect the host. At the same time, ACE2 is often abnormally expressed in non-small cell lung cancer. Therefore, understanding the respective mechanisms of ACE2 in COVID-19 and non-small cell lung cancer has extremely important reference value for the study of vaccines and therapeutic drugs, and also provides meaningful guidance for the protection of patients with lung cancer during the epidemic. This article reviews the possible invasive mechanism of ACE2 in SARS-CoV-2 and its abnormal expression in non-small cell lung cancer.