Regulation of pH on inflation and deflation of biosynthetic gas vesicles used as ultrasound molecular imaging probes / 生物工程学报

Gas vesicles (GVs) are gas-filled protein nanostructures that can regulate the buoyancy of microorganisms such as cyanobacteria and archaea. Recent studies have shown that GVs have the potential to be used as ultrasound molecular imaging probes in disease diagnosis and treatment. However, the mechanism of the inflation and deflation of GVs remains unclear, which hampers the preservation of GVs and gas replacement. In the present study, the environmental pH value was found to be an important factor in regulating the inflation and deflation of GVs. It can not only regulate the inflation and deflation of GVs in vivo to make Microcystis sp. cells present distinct levitation state, but also regulate the inflation and deflation of purified GVs in vitro, and the regulation process is reversible. Our results may provide a technical support for the large-scale production and preservation of biosynthetic ultrasound molecular imaging probes, especially for gas replacement to meet different diagnostic and therapeutic needs, and would facilitate the application of biosynthetic ultrasound molecular imaging probes.

Advances in mass-encoded probes for multiplex mass spectrometric detection / 中国药科大学学报

@#Mass-encoded probe is a probing tool that specifically identifies target molecules and thus outputs their characteristic ion signals with mass tags.It plays an important role in multiplex assay of disease markers, drug target screening and other biomedical applications.Based on various mass spectrometric methods, researchers have developed an array of mass tag-encoded probes with different structures and functions, providing powerful technical tools for multiplex detection of biomolecules in physiological environments and for mass spectrometry imaging of tissue samples.This review introduces the latest research progress of mass tag-encoded probes in multiplex mass spectrometric detection from three aspects, i.e. structural composition of the probes, mass spectrometric methods and their application in biochemical analysis, with a prospect of the future development of mass tag-encoded probes.

Colonization dynamics of subgingival microbiota in recently installed dental implants compared to healthy teeth in the same individual: a 6-month prospective observational study

Abstract Objectives To evaluate the colonization dynamics of subgingival microbiota established over six months around newly installed dental implants in periodontally healthy individuals, compared with their corresponding teeth. Methodology Seventeen healthy individuals assigned to receive single dental implants participated in the study. Subgingival biofilm was sampled from all implant sites and contralateral/ antagonist teeth on days 7, 30, 90, and 180 after implant installation. Microbiological analysis was performed using the Checkerboard DNA-DNA hybridization technique for detection of classical oral taxa and non-oral microorganisms. Significant differences were estimated by Mann-Whitney and Friedman tests, while associations between implants/teeth and target species levels were assessed by linear regression analysis (LRA). Significance level was set at 5%. Results Levels of some species were significantly higher in teeth compared to implants, respectively, at day 7 ( V.parvula , 6 × 10 5 vs 3 × 105 ; Milleri streptococci , 2 × 10 6 vs 6 × 10 5 ; Capnocytophaga spp., 2 × 10 6 vs 9 × 10 5 ; E.corrodens , 2 × 10 6 vs 5 × 10 5 ; N. mucosa , 2 × 10 6 vs 5 × 10 5 ; S.noxia , 2 × 10 6 vs 3 × 10 5 ; T.socranskii , 2 × 10 6 vs 5 × 10 5 ; H.alvei , 4 × 10 5 vs 2 × 10 5 ; and Neisseria spp., 6 × 10 5 vs 4 × 10 4 ), day 30 ( V.parvula , 5 × 10 5 vs 10 5 ; Capnocytophaga spp., 1.3 × 10 6 vs 6.8 × 10 4 ; F.periodonticum , 2 × 10 6 vs 10 6 ; S.noxia , 6 × 10 5 vs 2 × 10 5 ; H.alvei , 8 × 10 5 vs 9 × 10 4 ; and Neisseria spp., 2 × 10 5 vs 10 6 ), day 120 ( V.parvula , 8 × 10 5 vs 3 × 10 5 ; S.noxia , 2 × 10 6 vs 0; and T.socranskii , 3 × 10 5 vs 8 × 10 4 ), and day 180 ( S.enterica subsp. enterica serovar Typhi, 8 × 10 6 vs 2 × 10 6 ) (p<0.05). Implants showed significant increases over time in the levels of F.nucleatum , Gemella spp., H.pylori , P.micra , S.aureus , S.liquefaciens , and T.forsythia (p<0.05). LRA found that dental implants were negatively correlated with high levels of S. noxia and V. parvula (β=-0.5 to -0.3; p<0.05). Conclusions Early submucosal microbiota is diverse and only a few species differ between teeth and implants in the same individual. Only 7 days after implant installation, a rich microbiota can be found in the peri-implant site. After six months of evaluation, teeth and implants show similar prevalence and levels of the target species, including known and new periodontopathic species.

Recent advances in ultrasound-controlled fluorescence technology for deep tissue optical imaging / 药物分析学报

Fluorescence imaging is a noninvasive and dynamic real-time imaging technique;however,it exhibits poor spatial resolution in centimeter-deep tissues because biological tissues are highly scattering media for optical radiation.The recently developed ultrasound-controlled fluorescence(UCF)imaging is a novel imaging technique that can overcome this bottleneck.Previous studies suggest that the effective contrast agent and sensitive imaging system are the two pivotal factors for generating high-resolution UCF images ex vivo and/or in vivo.Here,this review highlights the recent advances(2015-2020)in the design and synthesis of contrast agents and the improvement of imaging systems to realize high-resolution UCF imaging of deep tissues.The imaging performances of various UCF systems,including the signal-to-noise ratio,imaging resolution,and imaging depth,are specifically discussed.In addition,the challenges and prospects are highlighted.With continuously increasing research interest in this field and emerging multidisciplinary applications,UCF imaging with higher spatial resolution and larger imaging depth may be developed shortly,which is expected to have a far-reaching impact on disease surveillance and/or therapy.

Application of organic fluorescent probe-assisted near infrared fluorescence imaging in cervical cancer diagnosis / 生物工程学报

Fluorescence imaging has been widely used in the fields of biomedicine and clinical diagnosis. Compared with traditional fluorescence imaging in the visible spectral region (400-760 nm), near-infrared (NIR, 700-1 700 nm) fluorescence imaging is more helpful to improve the signal-to-noise ratio and the sensitivity of imaging. Highly-sensitive fluorescent probes are required for high-quality fluorescence imaging, and the rapid development of nanotechnology has led to the emergence of organic dyes with excellent fluorescent properties. Among them, organic fluorescent probes with the advantages of high safety, good biocompatibility, and high optical stability, are more favorable than inorganic fluorescent probes. Therefore, NIR fluorescence imaging assisted with organic fluorescent probes can provide more structural and dynamic information of biological samples to the researchers, which becomes a hot spot in the interdisciplinary research field of optics, chemistry and biomedicine. This review summarizes the application of NIR organic fluorescent probes in cervical cancer imaging. Several typical organic fluorescent probes (such as indocyanine green, heptamethine cyanine dye, rhodamine and polymer fluorescent nanoparticles) assisted NIR fluorescence imaging and their applications in cervical cancer diagnosis were introduced, and the future development and application of these techniques were discussed.

The use of real time PCR for arboviruses diagnostics: integrative review / O uso de PCR em tempo real em diagnósticos de arboviroses: revisão integrativa

RESUMEN Las arbovirosis son enfermedades virales transmitidas por artrópodos (arthropod-borne virus). Dengue, zica y chikungunya se destacan entre los arbovirus emergentes y reemergentes en los últimos años en todo el mundo. La similitud de los síntomas de estas infecciones hace que el diagnóstico clínico sea ineficaz, dificultando las medidas profilácticas y preventivas para nuevos brotes. El diagnóstico molecular mediante la técnica de reacción en cadena de la polimerasa (PCR) en tiempo real es una de las formas de diagnosticar esas enfermedades. En este estudio se recopiló y evaluó la literatura sobre el diagnóstico de arbovirosis. Nuestro objetivo era responder a una pregunta orientadora: ¿la metodología de PCR en tiempo real es eficaz para diagnosticar arbovirosis? Se buscaron artículos científicos de acceso abierto en las bases de datos Pubmed (50 artículos) y Scielo (107 artículos), entre 2014 y 2019. La selección se realizó utilizando los criterios de inclusión y exclusión, quedando solo 20 artículos. Entre estos, el 85% fueron estudios transversales, el 10% fueron revisiones sistemáticas y el 5% fueron estudios de casos. El período de publicaciones fue del 50% en 2017; 35% en 2016; y 5% en 2014, 2015 y 2019, cada. En cuanto a los virus tratados en los artículos, el 25% de los estudios investigaron sobre el dengue; el 25% el chikungunya y el 20% el virus del Zica. La efectividad del diagnóstico molecular se publicó en el 21% de los artículos (sensibilidad y especificidad); el 53% destacó el límite de detección; 70%, ausencia de reacciones cruzadas; y el 80%, la diferenciación entre virus.

Accuracy and reproducibility of UNC-15 and Gram probe #2 in measuring probing pocket depths

@#<p style="text-align: justify;"><strong>Objective.</strong> Second-generation force-controlled periodontal probes were developed to address examiner-related factors in the application of standard forces while probing. However, previous researches comparing the accuracy of first and second-generation probes have failed to yield unequivocal results. This study aimed to determine the accuracy and reproducibility of a first-generation UNC-15 probe and force-controlled Gram probe #2, used by senior and junior dental students in measuring probing pocket depths (PPD).</p><p style="text-align: justify;"><strong>Methods.</strong> Eight senior and eight junior dental students participated in the study. A periodontal model was positioned on a digital balance. Each participant performed probing four times, twice using a UNC-15 and twice employing a Gram probe #2. The order of probe usage was randomly assigned. The PPD and probing forces employed were recorded initially and after a 15-minute interval. Data were analyzed using a t-test at a 0.05 significance level.</p><p style="text-align: justify;"><strong>Results.</strong> Both junior and senior students used significantly higher probing forces (35.3 ± 1.4 and 29.9 ± 2.1 g, respectively) than the standard 25 g when using the UNC-15 probe. The junior students employed accurate forces using the Gram probe while probing both anterior (25.6 ± 1.2 g) and posterior teeth (25.1 ± 0.7 g). In comparison, the seniors used accurate forces (26.0 ± 1.3 g) only while examining anterior teeth. The PPD values obtained by both student groups were comparable to the actual PPD, except for significantly higher measurements (P=0.0003) obtained by juniors when examining posterior teeth using the Gram probe. When assessing the reproducibility of PPD measurements obtained from two examinations, values were reproducible for both juniors and seniors when using the Gram probe and during an inspection of posterior teeth using the UNC-15 probe.</p><p style="text-align: justify;"><strong>Conclusions.</strong> The Gram probe #2 had better reproducibility and yielded forces closer to the standard 25 g. However, more accurate PPD values were obtained with the UNC-15 probe.</p>

Descubrimiento de fármacos basado en imagenología de células vivas / Drug discovery based on live cell imaging

RESUMEN El diseño eficiente de compuestos aprovechando las características estructurales de las moléculas y la búsqueda eficiente de dianas terapéuticas, ha proporcionado herramientas efectivas en la investigación de nuevos tratamientos cuando esta se enfoca en mecanismos celulares de la enfermedad. Los cambios fenotípicos producidos por la interacción in vitro entre molécula-diana, pueden controlarse cuantitativamente mediante imagenología de células vivas. Para garantizar una interacción adecuada, es necesario considerar diferentes elementos cruciales: 1. Las características estructurales y la dinámica molecular del compuesto a evaluar. 2. La relevancia del blanco para la fisiopatología de interés. Sin embargo, el desconocimiento del panorama general en el descubrimiento de fármacos, desde problemáticas estructurales y celulares, ha enlentecido la búsqueda de nuevos tratamientos. Esta revisión descriptiva de tema presenta algunos aspectos estructurales importantes para la caracterización de compuestos como candidatos terapéuticos, y aproximaciones experimentales para desarrollo de sistemas celulares. Los tópicos discutidos se enfocan en la monitorización por imagenología de células vivas y así mismo proporcionamos ejemplos relevantes. La monitorización de efectos fenotípicos producidos por interacciones entre candidato químico y blanco terapéutico en un sistema celular puede favorecer la búsqueda eficiente de moléculas potencialmente terapéuticas.

SUMMARY The efficient compounds' design taking advantage of the molecule's structural characteristics and efficient search for therapeutic targets has provided effective tools for the research of new treatments when this is focused on disease cellular mechanisms. Phenotypic changes produced by in vitro interaction between molecules and targets can be monitored quantitatively by live cell imaging. To guarantee adequate interaction, it is necessary to consider different crucial elements: 1. Structural characteristics and molecular dynamics of the evaluated compound. 2. Target relevance for the concern physiopathology. However, overview's ignorance of the drug discovery, from structural and cellular problems, has slowed the new treatments research. This literature review presents some important structural aspects for compounds' characterization as therapeutic candidates and experimental approaches for cellular systems development. Subjects discussed are focused on live cell imaging and we also provide relevant examples. Phenotypic monitoring of interactions' produced effects between the chemical candidate and therapeutic target in a cellular system can favor the efficient search of potentially therapeutic molecules.

Advances of near-infrared fluorescent probes for detection of Alzheimer′s disease / 中国药科大学学报

@#Alzheimer′s disease(AD)is a chronic neurodegenerative diseasecommonly seen in the elderlys. Several therapeutic drugs have failed in phase III clinical trials in recent years and there have been no efficient treatment for AD currently. Thus, there has been an urgent need for the effective methods of AD diagnosis at early stage. Developingnear-infrared fluorescentprobes for AD hallmarks detection has been a promising research field. In this review, we summarized a variety of near-infrared fluorescence(NIRF)probes reported in the past decade, which capable of detecting β-amyloid, Tau protein and reactive oxygen species, including their chemical strucutres、optical properties, in vitro and in vivo studies. Furthermore, we alsoraised some new directions for AD diagnosing. We believe that these new directions raised herein will benefit the future development of NIRF probes in the field of AD research.

In vitro multi-modal imaging of magnetic targeted nanoparticles and their targeting effect on hepatic stellate cells / 中国组织工程研究

BACKGROUND: In recent years, molecular imaging combined with medical imaging technology and targeted molecular probes have gradually become a research focus. The targeted tissues at the molecular level can be observed using molecular imaging, medical imaging technology, and targeted molecular probes in combination to realize non-invasive imaging of the occurrence and development of the diseases. OBJECTIVE: To develop the magnetic targeted nanoparticle probes, observe the ultrasound/CT/MRI imaging properties in vitro, and investigate their targeting ability to rat hepatic stellate cells in vitro. METHODS: Taking poly(lactic-co-glycolic acid) (PLGA) polymer as the shell, cyclic arginine-glycine-aspartic acid (cRGD) octapeptide as the ligand, targeted magnetic nanoparticles with superparamagnetic Fe3O4 embedded in the shell and perfluorooctyl bromide(PFOB) loaded in the core were prepared by double emulsion evaporation method. The physical and chemical properties of the nanoparticles were detected. The ultrasound/CT/MRI multi-modal imaging properties of the nanoparticles at different concentrations diluted with double-distilled water were tested in vitro. Cyclic RGD peptide immobilization on PLGA-Fe3O4-PFOB NPs was completed through the amide condensation reaction. The conjugation efficiency of the cRGD on PLGA-Fe3O4-PFOB NPs and targeting ability of targeted magnetic nanoparticles in vitro were verified. Cytotoxicity experiments were used to measure the toxic effects of nanoparticles at different concentrations on BRL-3A cells in each group. RESULTS AND CONCLUSION: The targeted magnetic nanoparticles with the average size of (221. 5±60. 3) nm were uniform in dispersion and size. The prepared individual nanoparticle was spherical with the superparamagnetic Fe3O4 scattered on the shell. The encapsulation rate of Fe3O4 was 38%. In vitro ultrasound imaging and CT imaging signal decreased gradually as the concentrations of the nanoparticle suspension decreased. The T2-weighted signal of MRI decreased gradually with the increase of the concentrations of magnetic particle Fe3O4. Flow cytometry results showed that 94. 13% of the cRGD was bound to the nanoparticles. In vitro cell targeting experiments showed that compared to PLGA-Fe3O4-PFOB NPs, cRGD-PLGA-Fe3O4-PFOB NPs exhibited greater cell targeting and affinity efficiency to hepatic stellate cells. Cytotoxicity experiments results showed the nanoparticles had no significant influence on cell viability of the BRL-3A cells. These results suggest that targeted magnetic nanoprobe cannot only be used as a multi-modal imaging contrast agent for ultrasound/CT/MRI, but also exhibits a strong specific affinity to rat hepatic stellate cells in vitro. It has great potential for the early diagnosis of liver fibrosis.

Analysis of full-length sequence and 18 point mutations of HLA-B in a leukemia patient and her family / 中国组织工程研究

BACKGROUND: The human leukocyte antigen (HLA) has undergone long-term evolution to form diverse polymorphisms. In recent years, due to the increase in the number of examinees and the rapid development of HLA typing technology, novel HLA alleles have been discovered constantly. OBJECTIVE: To analyze the full-length sequence and 18 point mutations of HLA-B gene in a leukemia patient and her family using the next-generation sequencing technology. METHODS: Polymerase chain reaction and sequence-specific oligonucleotide probes (PCR-SSOP) and polymerase chain reaction-sequence based typing (PCR-SBT) revealed abnormalities in the patient’s HLA-B. To identify the genotype, we sequenced the full length of the gene by next-generation sequencing technology and collected blood samples from the patient’s father, mother and two sisters for genetic analysis of HLA genes. RESULTS AND CONCLUSION: Both PCR-SSOP and PCR-SBT indicated that the HLA-B sample had no perfectly matched genotype. Further detection using the next-generation sequencing technology revealed that the novel allele had 18 base mutations in the exon, intron and 3’UTR compared to the most homologous allele B*15:09:01. Five exon base mutations were located in the exons 3 and 4, which were: 486G→C, 583T→C, 636T→C, 652A→G, 756C→T, resulting in changes in the five corresponding codons, including 171 tyrosine (Tyr) → histidine (His) and 194 isoleucine (Ile) → valine (Val). A pedigree survey found that the patient’s novel HLA B allele was inherited from her father. The novel allele sequence was submitted to the Genbank database (MG595995). A novel HLA-B allele was confirmed by the next-generation sequencing, which was officially named HLA-B*15:435 by the World Health Organization HLA Factor Nomenclature Committee in December 2017.

Advances in second near-infrared fluorescent nanoprobes-based in vivo imaging for biomedical applications / 上海交通大学学报（医学版）

Besides the advantages of high sensitivity, non-invasive, and real-time imaging that the general optical imaging technologies have, the second near-infrared (1 000-1 700 nm) in vivo imaging is regarded as one of the most promising optical imaging technologies in clinical applica-tion at present due to its advantages such as low autofluorescence background, deep tissue penetration and clear imaging. In the second near-infrared imaging system, the fluorescence probe is the most critical technical difficulty and the current research hotspot. Thereinto, due to the unique physical and chemical properties of nanomaterials, the second near-infrared fluorescent nanoprobe is an ideal contrast agent for the current second near-infrared optical imaging technology. The reported inorganic materials that can be used as the second near-infrared fluores-cent nanoprobes include quantum dots, rare earth nanoparticles, and single-walled carbon nanotubes, and the reported organic materials are mainly conjugated polymers. In this paper, the research progresses of the second near-infrared fluorescence nanoprobes mentioned above are re-viewed in order to better understand their application to the field of biomedicine, and promote the further research and clinical application of the second near-infrared in vivo imaging to the biomedical field.

Fluorescent probes for the detection of reactive oxygen species in human spermatozoa / 亚洲男科学杂志(英文版)

Reactive oxygen species (ROS) production is a by-product of mitochondrial activity and is necessary for the acquisition of the capacitated state, a requirement for functional spermatozoa. However, an increase in oxidative stress, due to an abnormal production of ROS, has been shown to be related to loss of sperm function, highlighting the importance of an accurate detection of sperm ROS, given the specific nature of this cell. In this work, we tested a variety of commercially available fluorescent probes to detect ROS and reactive nitrogen species (RNS) in human sperm, to define their specificity. Using both flow cytometry (FC) and fluorescence microscopy (FM), we confirmed that MitoSOX™ Red and dihydroethidium (DHE) detect superoxide anion (as determined using antimycin A as a positive control), while DAF-2A detects reactive nitrogen species (namely, nitric oxide). For the first time, we also report that RedoxSensor™ Red CC-1, CellROX®Orange Reagent, and MitoPY1 seem to be mostly sensitive to hydrogen peroxide, but not superoxide. Furthermore, mean fluorescence intensity (and not percentage of labeled cells) is the main parameter that can be reproducibly monitored using this type of methodology.

HPV infection and endometriosis: a systematic review and meta-analysis / Infecção por HPV e endometriose: uma revisão sistemática e metanálise

Introduction: Recent research has focused on the role of persistent ascending bacterial infections and sexually transmitted infections (STI) as a factor associated to endometriosis. Indeed, some studies investigated the possible role of HPV in endometriosis, but this topic remains inconclusive. Objective:The present study aims to meta-analyze research that assessed the presence of HPV infection in patients with endometriosis. Methods: MEDLINE, Embase, Scopus, LILACS, Cochrane Library, and OpenGrey were searched until February 10th, 2020. Search terms included "endometriosis" and "HPV" without language restrictions. The combined relative risks and 95% confidence interval (95%CI) were calculated, and heterogeneity was assessed with I-square (I2 ). Results: Meta-analysis with low heterogeneity found a relative risk of twice as much in women exposed to HPV in relation to the unexposed control. Conclusion: Results indicate that HPV could be a risk factor for developing endometriosis.

Introdução: A pesquisa recente enfocou o papel de infecções por bactéria ascendente persistente e infecções sexualmente transmissíveis (IST) como um dos fatores associados à endometriose. Na verdade, alguns estudos investigaram o possível papel do HPV na endometriose, mas esse tópico permanece inconclusivo. Objetivo: O presente estudo tem como objetivo fazer uma metanálise de pesquisas que avaliaram a presença de infecção por HPV em pacientes com endometriose. Métodos: As bases de dados MEDLINE, Embase, Scopus, LILACS, Biblioteca Cochrane e OpenGrey foram pesquisadas até 10 de fevereiro de 2020. Os termos de pesquisa incluíram "endometriose" e "HPV" sem restrições de idioma. Os riscos relativos combinados e intervalo de confiança de 95% (IC95%) foram calculados, e a heterogeneidade foi avaliada usando o I-quadrado (I2 ). Resultados: A metanálise com baixa heterogeneidade encontrou um risco relativo duas vezes maior em mulheres expostas ao HPV em relação ao controle não exposto. Conclusão: Os resultados indicam que o HPV pode ser um fator de risco para o desenvolvimento de endometriose.

Microbial profile and endotoxin levels in primary periodontal lesions with secondary endodontic involvement

Abstract This study was carried out to investigate the microbial profile and endotoxin levels of endodontic-periodontal lesions of periodontal origin. Periodontal and endodontic samples were taken from periodontal pockets and necrotic root canals of 10 teeth with endodontic-periodontal lesions. Evidencing of 40 different bacterial species were determined in each endodontic and periodontal sample using the checkerboard DNA-DNA hybridization method and Kinetic chromogenic LAL assay was used for quantification of endotoxins. Fisher's exact test correlated the bacterial species with the endodontic or periodontal microbiota. The endotoxin levels (EU/mL) found in samples of the root canal and periodontal pocket were compared by the Wilcoxon test (p<0.05). Bacteria and LPS units were found in 100% of the endodontic and periodontal samples. The species E. faecium, P. acnes, G. morbillorum, C. sputigena and L. buccalis were strongly correlated with the endodontic microbiota and P. nigrescens with the periodontal microbiota. P. intermedia, P. endodontalis and V. parvula were more prevalent in both endodontic and periodontal microbiots. The endotoxin levels in the periodontal pocket (89600 EU/mL) were significantly higher than in the root canal (2310 EU/mL). It was concluded that the microbiota present in the periodontal and endodontic tissues is similar, with a higher prevalence of species of the orange complex and a higher level of endotoxin in the periodontal pockets.

Resumo Este estudo foi realizado para investigar o perfil microbiano e os níveis de endotoxina de lesões endoperiodontais de origem periodontal. Amostras periodontais e endodônticas foram obtidas de bolsas periodontais e canais radiculares necróticos de 10 dentes com lesões endoperiodontais. A investigação de 40 espécies bacterianas diferentes foram determinadas em cada amostra endodôntica e periodontal usando o método de hibridização de DNA-DNA (checkerboard) e o ensaio cinético cromogênico LAL foi usado para quantificação de endotoxinas. O teste exato de Fisher correlacionou as espécies bacterianas com a microbiota endodôntica ou periodontal. Os níveis de endotoxina (EU/mL) encontrados nas amostras do canal radicular e na bolsa periodontal foram comparados pelo teste de Wilcoxon (p<0,05). Bactérias e unidades de LPS foram encontradas em 100% das amostras endodônticas e periodontais. As espécies E. faecium, P. acnes, G. morbillorum, C. sputigena e L. buccalis foram fortemente correlacionadas com a microbiota endodôntica e P. nigrescens com a microbiota periodontal. P. intermedia, P. endodontalis e V. parvula foram mais prevalentes em ambas microbiotas endodôntica e periodontal. Os níveis de endotoxina na bolsa periodontal (89600 EU/mL) foram significativamente maiores do que no canal radicular (2310 EU/mL). Concluiu-se que a microbiota presente nos tecidos periodontal e endodôntico é semelhante, com maior prevalência de espécies do complexo laranja e maior nível de endotoxina nas bolsas periodontais.

Targeting peptide-mediated 131I-polyamidoamine in medullary thyroid carcinoma bearing mice / 中华核医学与分子影像杂志

Objective@#To construct 131I-the fifth generation polyamidoamine (PAMAM(G5.0)) with targeting peptide Ser-Arg-Glu-Ser-Pro-His-Pro (SRESPHP; SR) or Gly-Pro-Leu-Pro-Leu-Arg (GPLPLR; GP) and double targeting peptide SR/GP, and evaluate the targeting ability in medullary thyroid carcinoma (MTC) model.@*Methods@#PAMAM(G5.0), PAMAM(G5.0)-SR, PAMAM(G5.0)-GP and PAMAM(G5.0)-SR/GP were radiolabeled with 131I by chloramine T method. The radiolabeled yield and radiochemical purity were determined by thin layer chromatography. MTC xenografts were developed and the percentage radio-activity of injection dose per gram of tissue (%ID/g) in tumor and organs was measured at 24 h post-injection. Region of interest (ROI) was drawn and the tumor/non-tumor (T/NT) ratios at 4, 8 and 24 h post-injection were calculated and compared among different groups. One-way analysis of variance, repetitive measurement analysis of variance and Dunnett-t test were used to compare the data of different groups. The relationship between %ID/g and T/NT was analyzed with Pearson correlation.@*Results@#The radiolabeled yield was more than 75% and radiochemistry purity was more than 90%. The difference of %ID/g at 24 h post-injection was significant (F=14.400, P<0.001) in tumors of all groups. The radioactive uptake in tumor of 131I-PAMAM(G5.0)-SR group was the highest at 24 h post-injection((1.80±0.18) %ID/g). There were significant differences of T/NT ratios among different groups(F=4.776, P<0.05)and between different time points(F=8.630, P<0.05). Compared with negative control group (Na131I), the T/NT ratios significantly increased in 131I-PAMAM(G5.0)-SR group at 4, 8 and 24 h post-injection (t=4.169, 7.123 and 4.032, all P<0.05) and in 131I-PAMAM(G5.0)-GP group at 4 h post-injection (t=5.893, P<0.05). The T/NT ratio in 131I-PAMAM(G5.0)-SR group was higher than that in 131I-PAMAM(G5.0)-GP group at 24 h post-injection (t=2.871, P<0.05).@*Conclusions@#PAMAM(G5.0)-SR, PAMAM(G5.0)-GP and PAMAM(G5.0)-SR/GP can target the MTC models. 131I-PAMAM(G5.0)-SR has the best biological properties and may provide a new precision method for MTC diagnosis, treatment and prognosis evaluation.

Environment-responsive fluorescence probes in image-guided surgery / 中华核医学与分子影像杂志

Unlike conventional imaging technologies, fluorescent imaging benefits from its safety, high-spatial resolution and real-time capability, which make it a highly adoptable imaging method for tumor detection and image-guided surgery in clinics. There are two types of fluorescent probes, including always-on type and environment-responsive type, wherein environment-responsive probes are preferred due to higher target-to-background ratios, which can improve sensitivity and specificity. The environment-responsive probes include enzyme-reactive probes, pH-sensitive probes and hypoxia responsive probes. This review summarizes recent progress in environment-responsive probes, and discusses their potentials in tumor detection and image-guided surgery.

Comparison of PCR-fluorescence probe and Xpert MTB／RIF in diagnosis of tuberculosis from non-respiratory specimens / 中华临床感染病杂志

Objective To compare the application of PCR-fluorescence probe, Bactec MGIT960 and Xpert MTB／RIF in diagnosis of tuberculosis from non-respiratory specimens.Methods Non-respiratory specimens from 225 patients with suspected tuberculosis admitted in Zhejiang Hospital of Integrated Chinese Medicine and Western Medicine from October 2017 to August 2018 were collected.There were 177 cases of tuberculosis and 48 cases of non-tuberculosis confirmed by clinical diagnosis.All specimens were tested with PCR-fluorescence probe, Xpert MTB／RIF and Bactec MGIT960.The clinical diagnostic results were used as the gold standard, and the receiver operating characteristic curve ( ROC) was drawn to evaluate the diagnostic values of three methods.The consistency of PCR-fluorescence probe method with Xpert MTB／RIF assay was analyzed.Results The sensitivity of PCR-fluorescent probe, Xpert MTB／RIF and Bactec MGIT960 in diagnosis of tuberculosis was 53.67%(95／177), 58.76%(104／177) and 31.07%(55／177), respectively.The sensitivity of PCR-fluorescent probe and Xpert MTB／RIF was higher than that of Bactec MGIT 960 culture ( χ2 ＝17.60 and 27.41, P＜0.01), while there was no significant difference between the PCR-fluorescent probe and the Xpert MTB／RIF (χ2 ＝0.93, P＞0.05).The specificity of three methods were 100.00%(48／48), 100.00%(48／48) and 97.92%(47／48), respectively (F＝1.83, P＞0.05).ROC curve analysis showed that the area under the ROC curve ( AUC) of PCR-fluorescent probe, Xpert MTB／RIF, and Bactec MGIT960 was 0.768, 0.794, and 0.645, respectively.The diagnostic value of PCR-fluorescent probe and Xpert MTB／RIF for tuberculosis was significantly higher than that of Bactec MGIT960 (Z＝5.19 and 6.52, P＜0.01); while Xpert MTB／RIF was superior to PCR-fluorescence probe (Z＝2.8, P＜0.05).In various types of specimens , there was no significant difference in the detection rate of tuberculosis between PCR-fluorescent probe method and Xpert MTB／RIF (χ2 ＝0.73, P＞0.05).The PCR-fluorescent probe and Xpert MTB／RIF had a good consistency (kappa＝0.829).Conclusion Xpert MTB／RIF is superior to PCR-fluorescence probe in the detection of tuberculosis in non-respiratory specimens such as tissues and pus, but the two have good consistency.The PCR-fluorescence probe method is economical and practical , and easy to promote, which has a high clinical application prospects.

Targeting peptide-mediated 131I-polyamidoamine in medullary thyroid carcinoma bearing mice / 中华核医学与分子影像杂志

Objective To construct 131 I-the fifth generation polyamidoamine (PAMAM(G5.0)) with targeting peptide Ser-Arg-Glu-Ser-Pro-His-Pro (SRESPHP;SR) or Gly-Pro-Leu-Pro-Leu-Arg (GPLPLR;GP) and double targeting peptide SR/GP,and evaluate the targeting ability in medullary thyroid carcinoma (MTC) model.Methods PAMAM(GS.0),PAMAM(GS.0)-SR,PAMAM(GS.0)-GP and PAMAM(GS.0)-SR/GP were radiolabeled with 131I by chloramine T method.The radiolabeled yield and radiochemical purity were determined by thin layer chromatography.MTC xenografts were developed and the percentage radio-activity of injection dose per gram of tissue (％ID/g) in tumor and organs was measured at 24 h post-injection.Region of interest (ROI) was drawn and the tumor/non-tumor (T/NT) ratios at 4,8 and 24 h post-injection were calculated and compared among different groups.One-way analysis of variance,repetitive measurement analysis of variance and Dunnett-t test were used to compare the data of different groups.The relationship between ％ID/g and T/NT was analyzed with Pearson correlation.Results The radiolabeled yield was more than 75％ and radiochemistry purity was more than 90％.The difference of ％lD/g at 24 h post-injection was significant (F=14.400,P＜0.001) in tumors of all groups.The radioactive uptake in tumor of 131I-PAMAM (G5.0)-SR group was the highest at 24 h post-injection ((1.80± 0.18) ％ID/g).There were significant differences of T/NT ratios among different groups (F =4.776,P＜ 0.05)and between different time points (F =8.630,P＜0.05).Compared with negative control group (Na131 I),the T/NT ratios significantly increased in 131I-PAMAM(G5.0)-SR group at 4,8 and 24 h post-injection (t=4.169,7.123 and 4.032,all P＜0.05) and in 131I-PAMAM(G5.0)-GP group at 4 h post-injection (t =5.893,P＜0.05).The T/NT ratio in 131I-PAMAM (G5.0)-SR group was higher than that in 131 I-PAMAM (G5.0)-GP group at 24 h post-injection (t=2.871,P＜0.05).Conclusions PAMAM(G5.0)-SR,PAMAM(G5.0)-GP and PAMAM(G5.0)-SR/GP can target the MTC models.131I-PAMAM(G5.0)-SR has the best biological properties and may provide a new precision method for MTC diagnosis,treatment and prognosis evaluation.

Clinicopathological characteristics of HPV+ oropharyngeal squamous cell carcinoma / 中华病理学杂志

Objective@#To observe the clinicopathologic features of oropharyngeal squamous cell carcinoma associated with human papilloma virus (OPSCC-HPV) and discuss the role and value of different in situ hybridization (ISH) detection methods for HPV in pathologic diagnosis.@*Methods@#Fifteen cases of OPSCC-HPV were collected from Department of Pathology, Beijing Tongren Hospital, Capital Medical University from January 2016 to August 2018. These cases were diagnosed in accordance with the WHO classification of head and neck tumors. The histopathologic features and the clinicopathologic data were retrospectively analyzed. Immunohistochemistry (two-step EnVision method) was done to evaluate the expression of p16, Ki-67 and p53. ISH was used to detect HPV DNA (6/11 and 16/18). RNAscope technology was used to evaluate the presence of HPV mRNAs (16 and 18).@*Results@#The mean age for the 15 patients (8 males, 7 females) was 47 years (range from 30 to 69 years). OPSCC-HPV typically presentedat an advanced clinical stage, six patients had cervical lymphadenopathy (large and cystic), seven had tonsillar swelling, one had tumor at base of tongue, and one had odynophagia. Microscopically the tumors exhibited distinctive non-keratinizing squamous cell carcinoma morphology. Cervical nodal metastases were large and cystic, with thickening of lymph node capsules. OPSCC-HPV raised from crypt epithelium and extended beneath the tonsillar surface epithelial lining as nests and lobules, often with central necrosis. Tumor cells displayed a high N: C ratio, and high mitotic and apoptotic rates. Tumor nests are often embedded within lymphoid stroma, and may be infiltrated by lymphoid cells.Fifteen cases (15/15) were strongly positive for p16; Ki-67 index were 60%-90%; they were focally positive or negative for p53. Ten cases (10/10) were negative for HPV 6/11 DNA, and one case(1/10) was focally positive for HPV16/18 DNA. Eleven cases (11/11) were strongly positive for HPV16 mRNA, one case was focally positive for HPV18 mRNA.@*Conclusions@#OPSCC-HPV is a pathologically and clinically distinct form of head and neck squamous cell carcinoma. OPSCC-HPV is associated with high-risk HPV (type 16) in all cases. Detection of high-risk HPV16 mRNA by RNAscope is of great significance in the final diagnosis and pathogen identification.