Role of micronucleus-activated cGAS-STING signaling in antitumor immunity / 浙江大学学报·医学版

cGAS-STING signaling is a significant component of the innate immune system and functions as a vital sentinel mechanism to monitor cellular and tissue aberrations in microbial invasion and organ injury. cGAS, a cytosolic DNA sensor, is specialized in recognizing abnormally localized cytoplasmic double-stranded DNA (dsDNA) and catalytically synthesizes the second messenger cyclic-GMP-AMP (cGAMP), which initiates a cascade of type I interferon and inflammatory responses mediated by STING. Micronucleus, a byproduct of chromosomal missegregation during anaphase, are also significant contributors to cytoplasmic dsDNA. These unstable subcellular structures are susceptible to irreversible nuclear envelope rupture, exposing genomic dsDNA to the cytoplasm, which potently recruits cGAS and activates STING-mediated innate immune signaling and its downstream activities, including type I interferon and classical nuclear factor-κB (NF-κB) signaling pathways lead to senescence, apoptosis, autophagy activating anti-cancer immunity or directly killing tumor cells. However, sustained STING activation-induced endoplasmic reticulum stress, activated chronic type I interferon and nonclassical NF-κB signaling pathways remodel immunosuppressive tumor microenvironment, leading to immune evasion and facilitating tumor metastasis. Therefore, activated cGAS-STING signaling plays a dual role of suppressing or facilitating tumor growth in tumorigenesis and therapy. This review elaborates on research advances in mechanisms of micronucleus inducing activation of cGAS-STING signaling and its implications in tumorigenesis and therapeutic strategies of malignant tumors.

Análisis preliminar de alimentos para la población vegana/vegetariana. ¿son buenos aportadores de minerales de interés nutricional? / Preliminary analysis of foods for the vegan/vegetarian population. Are they good suppliers of minerals of nutritional interest?

Resumen Introducción: recientemente, se ha evidenciado gran desarrollo de variados productos destinados a la población vegetariana/vegana. Sin embargo, su valor nutricional no ha sido estudiado en profundidad. El objetivo del presente estudio fue evaluar aporte potencial (AP) y porcentaje de cobertura de requerimientos diarios (%RD) de hierro, calcio y zinc de alimentos dirigidos a poblaciones adolescente y adulta vegetariana/vegana. Materiales y método: se analizaron cinco medallones (4 comerciales, 1 casero) y once bebidas (9 comerciales, 4 de ellas c/jugo frutal y 2 caseras) elaborados con materias primas vegetales. Se estableció el AP de Fe, Ca y Zn en los productos considerando su contenido y dializabilidad porcentual (D%). Se calculó, para una porción de alimento, el porcentaje de cobertura del requerimiento diario de estos minerales. Resultados: el contenido de los minerales en los medallones fue: [Fe] 1,64-4,21 mg%; [Ca] 104-213 mg% y [Zn] 0,53-3,57 mg%; en las bebidas se observó: [Fe] 0,24-2,39 mg%; [Ca] 71-214 mg% y [Zn] 0,18-0,79 mg%. La D% en medallones fue: Fe 7,3-11,9; Ca 10,2-18,2 y Zn 17,0-21,4 y para las bebidas, Fe 5,2-32,8; Ca 6,4-35,7 y Zn 5,9-33,9. El %RD para adolescentes, considerando una porción de medallones fue: Fe mujeres 3,0-9,0%; hombres 5,0-16%; Ca 0,7-6,8% y Zn 1,5-3,9% y para adultos fue: Fe mujeres 3,0-10%; hombres 7,0-20%; Ca 0,6-5,8% y Zn 2,0-5,1%. Al considerar las bebidas, el %RD para adolescentes fue: Fe mujeres 2,0-28%; hombres 4,0-53%; Ca 0,4-22% y Zn 1,3-9,5%. Para adultos fue: Fe mujeres 2,0-31%; hombres 5,0-64%; Ca 0,3-19% y Zn 1,8-12%. Conclusiones: en los medallones se observó bajo %RD para los minerales estudiados. Las bebidas con agregado de jugos de naranja o manzana aportaron cantidades significativas de hierro. El %RD para zinc y calcio de los dieciséis alimentos fue bajo (ambos grupos estudiados). Consecuentemente, para cubrir los requerimientos de estos minerales habría que combinar adecuadamente los alimentos que se consumen.

Abstract Introduction: recently, there has been great development of various products aimed at the vegetarian/vegan population. However, its nutritional value has not been studied in depth. The objective of this study was to evaluate potential intake (PI) and percentage of coverage of daily requirements (% RD) of iron, calcium and zinc from foods aimed at adolescent and adult vegetarian/vegan populations Materials and method: five medallions (4 commercial, 1 homemade) and eleven beverages (9 commercial, 4 of them with fruit juice and 2 homemade) made with vegetable raw materials were analyzed. The PI of Fe, Ca and Zn was established in the products considering their content and percentage dialyzability (D%). It was calculated, for a portion of food, the percentage of coverage of the daily requirement of these minerals. Results: the mineral content in the medallions was: [Fe] 1.64-4.21 mg%; [Ca] 104-213 mg% and [Zn] 0.53-3.57 mg%; in beverages it was observed: [Fe] 0.24-2.39 mg%; [Ca] 71-214 mg% and [Zn] 0.18-0.79 mg%. The D% in medallions was: Fe 7,3-11,9; Ca 10.2-18.2 and Zn 17.0-21.4 and for beverages, Fe 5.2-32.8; Ca 6.4-35.7 and Zn 5.9-33.9. The %RD for adolescents, considering a portion of medallions was: Fe women 3.0-9.0%; men 5.0-16%; Ca 0.7-6.8% and Zn 1.5-3.9% and for adults it was: Fe women 3.0-10%; men 7.0-20%; Ca 0.6-5.8% and Zn 2.0-5.1%. When considering beverages, the %RD for adolescents was: Fe women 2.0-28%; men 4.0-53%; Ca 0.4-22% and Zn 1.3-9.5%. For adults it was: Fe women 2.0-31%; men 5.0-64%; Ca 0.3-19% and Zn 1.8-12%. Conclusions: in the medallions, low % RD was observed for the minerals studied. Drinks with added orange or apple juices provided significant amounts of iron. The %RD for zinc and calcium of the sixteen foods was low (both groups studied). Consequently, to meet the requirements of these minerals, it would be necessary to properly combine the foods consumed.

Advances in cGAS-STING pathway and its inhibitors in immune and inflammatory diseases / 中国药理学通报

The cGAS-STING signaling pathway is one of the main pathways of immune defense against many types of pathogens. cGAS catalyzes the production of the second messenger cGAMP (cyclic GMP-tVMP) by recognizing plasma DNA and cGAMP subsequently binds to the interferon gene stimulating factor (STING). The pathway induces the production of type I interferon (IFN-I) and activates the innate immune system. The activation of the cGAS-STI]NG pathway could facilitate self-protection,thus STI]NG agonists for tumor immunotherapy have attracted much attention in recent years,and several drug candidates have been in clinical trials. Meanwhile,aberrant activation of cGAS-STI]NG could lead to autoimmune diseases and has attracted extensive interest in developing its inhibitors. This paper summarizes the mechanism and regulatory sites of the cGAS-STI]NG pathway,and outlines the research progress of cGAS-STING pathway-related immune and inflammatory diseases and its inhibitors.

Characterization of bacterial swarming motility: a review / 生物工程学报

Swarming motility is a typical synergistic motion, in which bacteria use flagella and Type Ⅳ Pili together to move collectively on semi-solid surfaces. Swarming motility is a hot topic of research in the field of microbiology because of its close relationship with biofilm formation, fruiting bodies formation, pathogen invasion and microbial dispersal and symbiosis. A large number of studies have been conducted on bacterial swarming motility, including changes in the expression of key proteins, changes in chemical communications between bacteria as well as mechanical changes. The expression of flagellin and the level of intracellular c-di-GMP complicatedly regulates the collective behavior of bacteria in colonies, which consequently impacts the swarming motility. The unique physical properties of swarmer cells are conducive to the expansion of the whole colony. Factors such as nutrient and water content in the surrounding growth environment of bacteria also affect the ability of bacteria to swarm to different degrees. It is challenging to construct a universal model of swarming motility based on the molecular mechanisms of swarming in the future.

Protein Containing the GGDEF Domain Affects Motility and Biofilm Formation in Vibrio cholerae and is Negatively Regulated by Fur and HapR / 生物医学与环境科学（英文）

OBJECTIVE@#This study aimed to investigate whether the VCA0560 gene acts as an active diguanylate cyclase (DGC) in Vibrio cholerae and how its transcription is regulated by Fur and HapR.@*METHODS@#The roles of VCA0560 was investigated by utilizing various phenotypic assays, including colony morphological characterization, crystal violet staining, Cyclic di-GMP (c-di-GMP) quantification, and swimming motility assay. The regulation of the VCA0560 gene by Fur and HapR was analyzed by luminescence assay, electrophoretic mobility shift assay, and DNase I footprinting.@*RESULTS@#VCA0560 gene mutation did not affect biofilm formation, motility, and c-di-GMP synthesis in V. cholerae, and its overexpression remarkably enhanced biofilm formation and intracellular c-di-GMP level but reduced motility capacity. The transcription of the VCA0560 gene was directly repressed by Fur and the master quorum sensing regulator HapR.@*CONCLUSION@#Overexpressed VCA0560 functions as an active DGC in V. cholerae, and its transcription is repressed by Fur and HapR.

Progress in DNA-sensing pathways in Epstein-Barr virus infection / 中华微生物学和免疫学杂志

Epstein-Barr virus (EBV) is generally susceptible in human beings and multi-organ systems can be involved in EBV infection, such as blood, respiratory, urinary, digestive and nervous systems. EBV infection also plays an important role in the pathogenesis of related tumors, autoimmune diseases and other diseases, posing a great threat to human health. As a DNA virus, EBV can be sensed by DNA recognition receptors to trigger a series of downstream immune responses. A DNA-sensing pathway consists of DNA sensors, adaptor molecules and downstream effector signals. Double-stranded DNA sensors mainly include absent in melanoma 2-like receptors (ALRs) and cyclic GMP-AMP synthase (cGAS). Adaptors were mainly stimulator of interferon genes (STING) and apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC). Downstream immune responses mainly involve typeⅠIFN, inflammasomes and proinflammatory cytokines. As a double-stranded DNA virus of the Herpesviridae family, EBV triggers complex innate and adaptive immune responses in the host, especially the sensing pathways mediated by a variety of DNA recognition receptors, which play a key role in host immune defense and pathogen immune evasion. This review made the DNA sensor as the clue to comprehensively summarize the progress in the activation, regulatory mechanism and clinical relevance of DNA-sensing pathways in EBV infection in recent years, aiming to achieve a better understanding of the host innate immune responses during EBV infection and provide an immunological basis for the prevention and treatment of EBV infection-related diseases.

QGIS Application for Spatial and Temporal Visualisation of Data for Food Safety Audits in Malaysia

@#Introduction: Advancement in digital technology opens new doors for food safety auditors when it comes to performing food safety audits. Surge of Covid cases since year 2020 has seen an unprecedented switch to remote auditing by the Food Safety and Quality Programme under the arm of Ministry of Health in Malaysia. Methods: This paper presents the use of QGIS, an open-source cross-platform for geographic information system (GIS) to store, manage and visualise 2 types of data, i.e. real time data collected via a mobile device using QField, an open-source mobile application and also fixed data retrieved from existing database. New data from obtained from field sampling and surveillance presents updated information for food safety auditing and enforcement purposes. A total of 4972 datasets were obtained from the Ministry of Health’s Food Safety and Quality Division database on food factories from all 13 states and 3 federal territories in Malaysia. These datasets were transformed and stored into QGIS point layer for performing data classification analysis on clustering of HACCP, GMP and MeSTI certifications. Results: The Penang state has the most HACCP certified companies in fish and fish product category, Selangor is the highest for confectionery industry and Sabah for food services. The general output of mobile GIS provides a big picture of distribution of food safety certifications in Malaysia while more specific adoption of QField can assist in effective field work planning for enforcement officers and auditors leading to cost calculation via information on location, distance and time. Conclusion: QGIS application for spatial and temporal visualisation of data benefits the food safety auditing in Malaysia

Analysis of global mRNA drug industrialization dynamics / 药学学报

The successful development and application of mRNA COVID-19 vaccine fully illustrated the great potential and application prospect of mRNA technology in the field of biomedicine. Currently, many companies worldwide are developing drugs and vaccines based on mRNA technology for the prevention and treatment of various diseases. It can be foreseen that with the continuous launch of mRNA drugs, commercial GMP production capacity matching them is also urgent. The optimization of production processes, intelligent manufacturing and other risk control strategies, as well as the control of industrialization costs, will help improve the core competitiveness of mRNA innovative drug development. In view of this, this article will provide an overview of the global production process of mRNA drugs and the progress of related GMP production dynamics, sort out the key chain points of the mRNA industry chain, explore the construction of the mRNA pharmaceutical enterprise value chain and the formation of core competitiveness, and provide reference and reference for the research and development of innovative mRNA drugs and high-quality development in China.

Launching a CDMO in Brazil aiming to develop biopharmaceuticals for clinical trials

The innovation timeline is expensive, risky, competitive, time-consuming, and labor-intensive. In order to overcome such challenges and optimize financial resources, pharmaceutical companies nowadays hire contract development and manufacturing organizations (CDMO) to help them. Based on the experience acquired first from the development of two biopharmaceuticals, the Heterologous Fibrin Sealant and the Apilic Antivenom, and more recently, during their respective clinical trials; the Center for the Study of Venoms and Venomous Animals (CEVAP) proposed to the Ministry of Health the creation of the first Brazilian CDMO. This groundbreaking venture will assist in converting a candidate molecule - from its discovery, proof of concept, product development, up to pilot batch production - into a product. The CDMO impact and legacy will be immense, offering service provision to the public and private sector by producing validated samples for clinical trials and academic training on translational research for those seeking a position in pharmaceutical industries and manufacturing platforms.(AU)

Expressão e localização celular da diguanilato ciclase DgcP em Pseudomonas aeruginosa / Expression and cell localization of the diguanylate cyclase DgcP in Pseudomonas aeruginosa

A bactéria Gram-negativa Pseudomonas aeruginosa é um patógeno oportunista frequentemente associado a vítimas de queimaduras graves ou indivíduos com fibrose cística, sendo os isolados resistentes a carbepenêmicos dessa espécie considerados pela OMS como uma das maiores ameaças ao controle de infecções. O estabelecimento da infecção por esse patógeno é dependente de uma série de fatores de virulência, entre eles o pilus tipo IV (T4P), que possui papel importante na adesão a superfícies e motilidade do tipo twitching, essenciais para a colonização do hospedeiro. Uma das moléculas importantes na diferenciação entre as formas séssil e planctônica de P. aeruginosa é o segundo mensageiro bis-(3,5)-di-guanosina monofosfato cíclico (c-di-GMP), cuja síntese é feita enzimaticamente por diguanilato ciclases (DGCs). DgcP é uma DGC localizada nos polos da célula, que tem sua atividade de síntese de c-di-GMP aumentada na presença da proteína FimV, essencial para a montagem do T4P em P. aeruginosa. Neste trabalho, ensaios de microscopia de fluorescência, organização e expressão gênica foram realizados com o objetivo de aumentar a compreensão sobre o papel de DgcP em relação a sua expressão e aos fatores que regulam o T4P de P. aeruginosa. A proteína DgcP em fusão com mNeonGreen no C-terminal, expressa a partir do locus cromossômico, se localiza de maneira predominantemente bipolar tanto na linhagem selvagem quanto nos mutantes ΔpilA, ΔpilR e ΔchpA, evidenciando que seu padrão de localização não depende dos sistemas de regulação Pil-Chp e PilS-PilR. Ensaios de RT-PCRmostraram que dgcP se encontra em operon com PA14_72430 e dsbA1, indicando um papel celular conjunto entre esses genes, até o momento, desconhecido. Por fim, ensaios de qRT-PCR revelaram que os níveis de mRNA de dgcP são invariáveis nas linhagens WT, ΔpilA, ΔpilR, ΔchpA e ΔfimV, cultivadas em meio líquido ou meio sólido. Os resultados aqui mostrados, combinados com trabalhos prévios do nosso e de outros grupos, sugerem que DgcP é uma diguanilato ciclase responsável por geração constante de c-di-GMP nos polos da célula, possivelmente, atuando na sinalização local dependente do dinucleotídeo cíclico, cuja localização e atividade não são dependentes dos sistemas de regulação que atuam sobre o T4P

The Gram-negative bacterium Pseudomonas aeruginosa is an opportunistic pathogen often associated with severe burn victims or individuals with cystic fibrosis, which carbapenem-resistant isolates were classified by th World Health Organization classified one of the greatest threats to infection control. The establishment of infection by this pathogen is dependent on a series of virulence factors, including the type IV pilus (T4P), which plays an important role in adhesion to surfaces and twitching motility, essential features for host colonization. Bis-(3',5')-cyclic dimeric guanosine monophosphate (c-di-GMP) is a second messenger that involved in processes of biofilm formation, motility, and virulence. The diguanylate cyclase DgcP synthetizes cdi-GMP and it is located at the cell poles, and its activity depends on the scaffold protein FimV, essential for T4P assembly in P. aeruginosa. By increasing c-di-GMP levels, DgcP decreases flagellum-dependent motility and increases biofilm formation. In this work, fluorescence microscopy, gene organization and expression assays were performed to understand the whether DgcP localization and expression are under the control of T4P regulatory proteins. Fluorescence microscopy analysis showed that DgcP localizes predominantly at both cell poles in ΔpilA, ΔpilR, and ΔchpA mutants, showing that its localization pattern does not depend on the Pil-Chp and PilS-PilR systems. Furthermore, RT-PCR assays showed that dgcP is found in an operon with PA14_72430 and dsbA1, indicating an unknown putative related cellular role for these genes. Finally, qRT-PCR assays indicated that DgcP expression is invariant in ΔpilA, ΔpilR, ΔchpA, and ΔfimV mutants, either in liquid or solid medium. The results shownhere, combined with previous work by ours and other groups, suggest that DgcP is a diguanylate cyclase responsible for constant generation of c-di-GMP at the cell poles, possibly acting in local signaling dependent on the cyclic dinucleotide, but that is not under the control of the known T4P regulatory systems

A review article on cleaning validation in pharmaceutical industry / Монголын эм зүй, эм судлал

Abstract@#In pharmaceutical industry there are some possibilities of contamination and cross contamination because of improper cleaning of equipment, apparatus, processing area or the starting material, this can lead to severe hazards, therefore in pharmaceutical industry we could not afford any contamination as well as cross contamination. This can be minimized by proper cleaning of equipment, apparatus as well as the processing area. Prevention of cross contamination is one of the most significant conditions of Good Manufacturing Practices for drugs. This is especially topical for a multipurpose (shared) manufacture where several medicinal products, including drugs of different pharmacotherapeutic groups, are produced using the same facilities (manufacturing areas, workrooms, and equipment). The industry is able to achieve these key goals with the help of implementation of GMP. Therefore, a perfect cleaning method is required for avoiding the possibilities of contamination and cross contamination, for this a validated program is required, this program is known as cleaning validation. “Cleaning validation is documented evidence which assure that cleaning of equipment, piece of equipment or system will obtain pre-determined and acceptable limits”.

ToxR represses the synthesis of c-di-GMP in Vibrio parahaemolyticus / 生物工程学报

Vibrio parahaemolyticus, the main pathogen causing seafood related food poisoning worldwide, has strong biofilm formation ability. ToxR is a membrane binding regulatory protein, which has regulatory effect on biofilm formation of V. parahaemolyticus, but the specific mechanism has not been reported. c-di-GMP is an important second messenger in bacteria and is involved in regulating a variety of bacterial behaviors including biofilm formation. In this study, we investigated the regulation of ToxR on c-di-GMP metabolism in V. parahaemolyticus. Intracellular c-di-GMP in the wild type (WT) and toxR mutant (ΔtoxR) strains were extracted by ultrasonication, and the concentrations of c-di-GMP were then determined by enzyme linked immunosorbent assay (ELISA). Three c-di-GMP metabolism-related genes scrA, scrG and vpa0198 were selected as the target genes. Quantitative real-time PCR (q-PCR) was employed to calculate the transcriptional variation of each target gene between WT and ΔtoxR strains. The regulatory DNA region of each target gene was cloned into the pHR309 plasmid harboring a promoterless lacZ gene. The recombinant plasmid was subsequently transferred into WT and ΔtoxR strains to detect the β-galactosidase activity in the cellular extracts. The recombinant lacZ plasmid containing each of the target gene was also transferred into E. coli 100λpir strain harboring the pBAD33 plasmid or the recombinant pBAD33-toxR to test whether ToxR could regulate the expression of the target gene in a heterologous host. The regulatory DNA region of each target gene was amplified by PCR, and the over-expressed His-ToxR was purified. The electrophoretic mobility shift assay (EMSA) was applied to verify whether His-ToxR directly bound to the target promoter region. ELISA results showed that the intracellular c-di-GMP level significantly enhanced in ΔtoxR strain relative to that in WT strain, suggesting that ToxR inhibited the production of c-di-GMP in V. parahaemolyticus. qPCR results showed that the mRNA levels of scrA, scrG and vpa0198 significantly increased in ΔtoxR strain relative to those in WT strain, suggesting that ToxR repressed the transcription of scrA, scrG and vpa0198. lacZ fusion assay showed that ToxR was able to repress the promoter activities of scrA, scrG and vpa0198 in both V. parahaemolyticus and E. coli 100λpir. EMSA results showed that His-ToxR was able to bind to the regulatory DNA regions of scrA and scrG, but not to the regulatory DNA region of vpa0198. In conclusion, ToxR inhibited the production of c-di-GMP in V. parahaemolyticus via directly regulating the transcription of enzyme genes associated with c-di-GMP metabolism, which would be beneficial for V. parahaemolyticus to precisely control bacterial behaviors including biofilm formation.

Validation of a method employing liquid chromatography to identify and quantify glycomacropeptide of casein in raw milk / Validación de un método mediante cromatografía líquida para identificar y cuantificar glicomacropéptido de caseína en la leche cruda

Background: The information on official testing methods, or regulatory methods in Colombia to test whey in milk is limited; this restriction of information goes against the possibility of mitigating the risk of food fraud. Objectives: The validation of an HPLC method to determine casein glycomacropeptide (c-GMP), a protein that countries such as Brazil, Spain, and Ecuador have used as an indicator of raw milk adulteration with whey, was carried out. Methods: A 10mL sample of raw milk is precipitated with 24% TCA using ultrasound, a process followed by filtration. The collected fraction ensured the separation of c-GMP and then injected into the liquid chromatography. Results: A 30 minutes analysis allowed the determination of c-GMP with a retention time of 12.9 ± 0.5 minutes. The performance characteristics method in the validation exercise were: recovery percentage 99.97%, linearity R2> 0.95; % RSD accuracy <5.3%. Conclusion, the method exhibits desirable attributes for the intended purpose

Antecedentes: En Colombia la información de dominio público en metodologías de análisis de lactosuero en leche es limitada, restringiendo la posibilidad de acceder a ellas para mitigar el riesgo de fraude alimentario. Objetivos: Se realizó validación de un método por HPLC para determinar en leche cruda c-GMP, proteína usada como indicador de adulteración en países como Brasil y Ecuador. Metodos: Una muestra de 10mL de leche cruda es precipitada con TCA al 24% empleando ultrasonido, proceso seguido por filtración. La fracción recolectada aseguró la separación del c-GMP para luego inyectar al cromatógrafo líquido. Resultados: La determinación de c-GMP permitió el análisis en 30 minutos con tiempo de retención de 12,9 ± 0,5 minutos. Las características de desempeño del método en el ejercicio de validación fueron: porcentaje de recuperación 99,97%, linealidad R2>0,95; precisión %RSD< 5,3%. Conclusión: el método al final del ejercicio exhibe atributos para el fin previsto

Effect of HBsAg on the production of interferon-α in peripheral blood plasmacytoid dendritic cells induced by the stimulator of interferon genes signaling pathway / 临床肝胆病杂志

ObjectiveTo investigate the effect of HBsAg on the expression of interferon-α (IFN-α) in peripheral blood plasmacytoid dendritic cells (pDCs) induced by the stimulator of interferon genes (STING) signaling pathway activated by cyclic GMP-AMP (cGAMP). MethodPeripheral venous blood was collected from healthy adults and the patients with chronic hepatitis B virus (HBV) infection who attended the outpatient service or were hospitalized in Department of Infectious Diseases, The Affiliated Hospital of Xuzhou Medical University, from February to December 2016, and peripheral blood mononuclear cells (PBMCs) were isolated and extracted. After the STING agonist cGAMP was added to PBMCs, ELISA was used to measure the levels of IFN-α, interferon-β, and tumor necrosis factor-α in supernatant. PBMCs from healthy adults were pre-incubated with HBsAg and then stimulated by cGAMP, and supernatant was collected to measure IFN-α. The magnetic-activated cell sorting method was used to remove pDCs from PBMCs, and after culture with cGAMP, ELISA was used to measure the level of IFN-α in supernatant. PBMCs from healthy adults were stimulated by HBsAg and/or cGAMP, and then flow cytometry was used to measure the frequency of pDCs. The independent samples t-test was used for comparison of continuous data between two groups. ResultsPBMCs from the patients with chronic HBV infection stimulated by cGAMP in vitro had a significantly lower level of IFN-α than healthy controls (469.72±18.95 vs 599.90±84.06, t=4.868, P=0.001). PBMCs from healthy adults co-cultured with HBsAg and stimulated by cGAMP had a significantly lower level of IFN-α than those in the non-HBsAg group (448.5±52.0 vs 571.0±30.8, t=4.500, P=0.011). Compared with PBMCs containing pDCs, PBMCs without pDCs stimulated by cGAMP had a significant reduction in the level of IFN-α (164.50±40.73 vs 339.50±35.33, t=6.482, P=0.001). Compared with PBMCs from healthy adults stimulated by cGAMP, PBMCs pre-incubated with HBsAg and then stimulated by cGAMP had a significant reduction in the frequency of pDCs (0.12%±0.04% vs 0.24%±0.04%, t=5.176, P=0.014). ConclusionHBsAg can inhibit the expression of IFN-α induced by the STING pathway in pDCs activated by cGAMP.

Analysis of Risk in Production of MRI and Some Suggestions for Field Inspection / 中国医疗器械杂志

In recent years, with the wide application of magnetic resonance imaging (MRI) equipment in clinical practice, the quality of the equipment causes adverse events, which put pressure on manufacturers, at the same time, it puts forward higher requirements for medical device supervisors. In order to help the medical device supervisors to clarify the key points of verification, this paper analyzes the main risk points in the production process of the product according to the medical device good manufacturing practice(GMP), and puts forward the suggestions for field verification, which has practical significance for the submission of verification efficiency.

Regulatory Study on Air Purification System of Medical Device Manufacturers / 中国医疗器械杂志

Objective Through regulatory study on the common defects of air purification system, this paper provides valuable reference for practitioners in medical device industry. Methods More than 100 verification results of different companies had been collected during 2015 to 2018, followed by systematically analysis of the defects related to air purification system. Result 70 types of common defects in 13 areas had been summarized, and 20 key points in verification had been briefly concluded. Conclusion Recognizing and understanding these summarized defects and key points will not only promote the unification of criterion scale, but also benefit enterprises for themselves, inspection, quality management improvement, and the plant transformation as well.

The Effect of Tadalafil on Renal Fibrosis Induced by Ureteral Obstruction / Efecto del Tadalafil en la Fibrosis Renal Inducida por Obstrucción Uretral

ABSTRACT Objective: It has been reported that phosphodiesterase-5 (PDE-5) inhibitors improve kidney function during acute and chronic renal failure. This study aimed to determine the possible therapeutic effects of tadalafil, a specific PDE-5 inhibitor, on renal fibrosis induced by unilateral ureteral obstruction (UUO). Methods: Male Sprague-Dawley rats were used and randomly divided into three groups (n = 6) as sham-operated, UUO and tadalafil-treated (10 mg/72 hours, ig) UUO (UUO+T) groups. Unilateral ureteral obstruction was induced by complete ligation of the left ureter and 14 days after surgery creatinine clearance, urinary cyclic guanosine monophosphate (cGMP), renal alpha-smooth muscle actin (α-sma) and transforming growth factor βeta (TGF-β) levels, as well as histologic changes, were observed in all the animals. Results: Unilateral ureteral obstruction-induced renal fibrosis was confirmed by increased α-sma level, collagen deposition, tubular dilation, inflammatory cell infiltration and necrosis. An increased renal TGF-β level and decreased urinary cGMP level was also observed in obstructed animals in addition to reduced creatinine clearance. Tadalafil treatment, which restored the animals 'urinary cGMP level, significantly attenuated the fibrotic changes and TGF-β increase in their kidneys. Conclusion: This study suggests that tadalafil treatment ameliorates renal fibrosis by reducing TGF-β expression and may have important clinical relevance since tadalafil is currently used clinically to treat erectile dysfunction and pulmonary hypertension.

RESUMEN Objetivo: Se ha reportado que los inhibidores de la fosfodiesterasa-5 (PDE-5) mejoran las funciones renales durante la insuficiencia renal aguda y crónica. Este estudio tuvo por objetivo determinar los posibles efectos terapéuticos del tadalafil - un inhibidor específico de la PDE-5 - sobre la fibrosis renal inducida por una obstrucción ureteral unilateral (OUU). Métodos: Se utilizaron ratas machos Sprague-Dawley, divididas de manera aleatoria en tres grupos (n = 6): operación simulada, OUU y tratamiento con tadalafil (10 mg/72 horas, IG), y OUU (OUU+T). La obstrucción uretral unilateral fue inducida por una ligadura completa del uréter izquierdo y 14 días después de la cirugía, se observaron niveles de monofosfato de guanosina cíclico (GMP) urinario, alfa-actina de músculo liso (α-SMA), y factor de crecimiento transformante βeta (FCT-β), así como cambios histológicos en todos los animales. Resultados: La fibrosis renal inducida por obstrucción uretral unilateral fue confirmada por un aumento del nivel de α-SMA, deposición de colágeno, dilatación tubular, infiltración de células inflamatorias y necrosis. También se observó un aumento del nivel de FCT-β renal y una disminución del nivel de GMP urinario en los animales con obstrucción, además de una reducción del aclaramiento de la creatinina. El tratamiento con tadalafil, que restauró el nivel de GMP urinario de los animales, atenuó significativamente los cambios fibróticos y el aumento de FCT-β en los riñones. Conclusión: Este estudio sugiere que el tratamiento con tadalafil mejora la fibrosis renal al reducir la expresión de FCT-β y puede tener una importante relevancia clínica por cuanto el tadalafil se usa hoy día clínicamente para tratar la disfunción eréctil y la hipertensión pulmonar.

Analysis of Risk in Production for PET/MR and Suggestions for Field Inspection / 中国医疗器械杂志

PET/MR is a high-end medical imaging equipment with integrating PET and MR equipment into the highly sophisticated one and has rich clinical and molecular diagnosis functions, can obtain comprehensive information about the human body structure, function and metabolism, is of great value for the diagnosis and treatment of disease improvement. In this paper, through the analysis of existing production risk points on one of the primary stages of the whole product life cycle, combining with the medical device good manufacture practice, some suggestions have been put forward exploratively to field inspection for PET/MR manufacturers. It has certain significance for regulators of medical devices to clear the production risk point and improve verification efficiency during field inspection.

Roles of signaling molecules in biofilm formation / 生物工程学报

Bacterial biofilm refers to a tunicate-like biological group composed of polysaccharide, protein and nucleic acid secreted by bacteria on the surface of the mucous membrane or biological material. The biofilm formation is a major cause of chronic infections. Bacteria could produce some secondary metabolites during the growth and reproduction. Some of them act as signaling molecules allowing bacteria to communicate and regulate many important physiological behaviors at multiple-cell level, such as bioluminescence, biofilm formation, motility and lifestyles. Usually, these signal molecules play an important role in the formation of bacterial biofilm. We review here the effects of related signal molecules of Quorum Sensing, cyclic diguanylate, Two-Component Systems and sRNA on the biofilm formation. Focusing on these regulation mechanism of signal molecules in the process of biofilm formation is necessary for the prevention and treatment of some chronic diseases.

Problem Analysis and Regu latory Countermeasures of Unannounced Inspection and Tracking Inspection of 28 Pharmaceutical Manufacturers / 中国药房

OBJECTIVE： To provide reference for pharmaceutical manufacturers improving the quality system of GMP and drug regulatory departments improving their supervision level. METHODS： Through analyzing and summarizing the problems existing in the 28 pharmaceutical enterprises which had been published on the website in the National Medical Products Administration from February 6th， 2018 to January 25th， 2019， the common problems were found and their causes were analyzed， then the regulatory countermeasures were put forward. RESULTS & CONCLUSIONS： Pharmaceutical enterprises have some problems of inadequate implementation of GMP， such as the inadequate performance of personnel in key positions and the unsatisfactory training effect of relevant personnel， the inconsistency between actual production technology and approved legal technology， the non-standard management of enterprise materials， the incomplete batch production records and the inability to effectively monitor the production cycle. However， there are also some problems in the supervision department， such as the large difference in the scale of inspectors’ on-site inspection， the need to strengthen the inspectors’ inspection ability and level， and the lack of innovation in the means of supervision. It is suggested that pharmaceutical manufacturers should improve the construction of GMP quality management system and strengthen the training of relevant personnel； the regulatory authorities should continue to promote the reform of “release， control and service”， strictly enforce the access conditions of inspectors， strengthen the training of inspectors and ideological construction of the inspector team，further strengthen the construction of supervision system and enhance the innovation of supervision means， so as to jointly maintain the safety， effectiveness and quality controllability of medicines.