Sobre o lugar e a função do fantasma entre os anos de 1953 a 1964 do ensino de Lacan / Sur la place et la fonction du fantôme entre les années 1953 et 1964 dans l'enseignement de Lacan / About the place and function of the phantasm between the years of 1953 to 1964 of Lacan's teaching

O conceito de fantasma possui uma importância crucial para o enquadramento da experiência em psicanálise. Este artigo pretende situar o lugar e a função do fantasma neurótico na experiência analítica partindo da função da fala, tal como proposto por Lacan. Para tanto, selecionamos alguns textos e seminários de Lacan situados entre os anos de 1953 a 1964. Nossa hipótese de trabalho consiste na suposição de que, ao enquadrar uma realidade constituída inconscientemente enquanto resposta ao desejo do Outro, o fantasma serve para perpetuar ao neurótico um senso de ser idêntico a si mesmo à medida em que tende a promover um apagamento de contradições provenientes da posição que, como sujeito, ocupa em relação a este desejo. Ao ser concebida como irrupção de um objeto com valor de não-eu no enquadre fantasmático, a angústia produz um efeito reverso, constituindo, por conseguinte, tanto uma contraprova quanto um apoio na demonstração dessa hipótese.

Le concept de fantôme revêt une importance cruciale pour encadrer l'expérience en psychanalyse. Cet article veut situer la place et la fonction du fantasme névrotique dans l'expérience analytique à partir de la fonction de la parole, comme le propose Lacan. Pour cela, nous avons sélectionné quelques textes et séminaires de Lacan situés entre les années 1953 et 1964. Notre hypothèse de travail est qu'en encadrant une réalité inconsciemment constituée en réponse au désir de l'Autre, le fantôme sert à perpétuer chez le névrosé un sentiment d'être identique à lui-même dans la mesure où il tend à favoriser un effacement des contradictions nées de la position qu'il occupe, en tant que sujet, par rapport à ce désir. Conçue comme l'irruption d'un objet ayant valeur de non-moi dans le cadre fantasmatique, l'angoisse produit un effet inverse, constituant donc à la fois une contre-preuve et un support à la démonstration de cette hypothèse.

The concept of the phantasm holds a crucial importance for framing the experience in psychoanalysis. This article aims to situate the place and function of the neurotic phantasm in the analytical experience, starting from the function of speech as proposed by Lacan. To do so, we have selected some texts and seminars by Lacan between the years of 1953 to 1964. Our working hypothesis is that by framing an unconsciously constituted reality as a response to the Other's desire, the phantasm serves to perpetuate in the neurotic subject a sense of being identical to themselves as it tends to promote an erasure of contradictions arising from the subject's position in relation to this desire. When conceived as the irruption of an object with a non-ego value in the phantasmatic frame, anxiety produces a reverse effect, thereby constituting both a counterproof and a support in demonstrating this hypothesis.

Periradicular and Peri-Implant Lesions Associated with Apical Impact of Osseointegrated Implants in Tooth Roots: Clinical Case Reports

This clinical case report aims to describe the development of periradicular and perimplant cystic lesions resulted from the intimate contact of the apical region of osseointegrated implants of dental roots, and discuss the reasons fo r failure of the guided bone regeneration procedure associated with platelet rich fibrin and leukocytes, this process was used to the treatment of the first case. Case Reports. Three cases were reported, two cases described the close contact between the tooth roots and the osseointegrated implants and another with a distance of 1.08 mm. All cases realized a radiographic, and they had not periapical lesions before contact with the apical region of the osseointegrated implants on the roots of the teeth. In the case with the largest cystic extension, the procedure was: removal of the osseointegrated implant with apicectomy of the neighboring teeth, excisional biopsy of the lesion, and grafting using the technique of guided bone regeneration associated with L-PRF. All three cases, endodontic treatment was performed on the neighboring teeth within 2 years of survival of the osseointegrated implants in order to reverse the existing lesion. Results. The diagnostic hypothesis of the three cases was periradicular and peri-implant lesion, arising from a contact of the apical region of the osseointegrated implant with the adja cent tooth. The distance of 1.08 mm between the apices did not ensure normality of the periradicular and peri-implant tissues. The intimate contact caused lesions of different extents and root fractures. Conclusion. Premature contact of the osseointegrated implant with the root region of the neighboring tooth may lead to the development of periradicular and peri-implant lesions, suggesting that it is not possible to control this infectious process with endodontic treatment of the injured tooth.

Este reporte de caso clínico tuvo como objetivo describir el desarrollo de lesiones quísticas perirradiculares y periimplantarias resultantes del contacto íntimo de la región apical de implantes osteointegrados de raíces dentales, y además discutir las razones del fracaso del procedimiento de regeneración ósea guiada asociado a fibrina rica en plaquetas y leucocitos. Este proceso se utilizó para el tratamiento del primer caso. Se reportaron tres casos, en dos casos se describieron el estrecho contacto entre las raíces de los dientes y los implantes osteointegrados y en el otro se determinó una distancia de 1,08 mm. En los tres casos se realizó una radiografía y se determinó que no existían lesiones periapicales, antes del contacto con la región apical de los implantes osteointegrados, en las raíces de los dientes. En el caso de mayor extensión quística, el procedimiento fue: extracción del implante osteointegrado con apicectomía de los dientes vecinos, biopsia excisional de la lesión e injerto mediante la técnica de regeneración ósea guiada asociada a L-PRF. En los tres casos, el tratamiento de endodoncia se realizó en los dientes vecinos dentro de los 2 años de supervivencia de los implantes osteointegrados para revertir la lesión existente. La hipótesis diagnóstica de los tres casos fue lesión perirradicular y periimplantaria, originada por un contacto de la región apical del implante osteointegrado con el diente adyacente. La distancia de 1,08 mm entre los ápices no aseguraba la normalidad de los tejidos perirradiculares y periimplantarios. El contacto íntimo provocó lesiones de diferente extensión y fracturas radiculares. El contacto prematuro del implante osteointegrado con la región radicular del diente vecino puede conducir al desarrollo de lesiones perirradiculares y periimplantarias, lo que sugiere que no es posible controlar este proceso infeccioso con tratamiento endodóntico del diente lesionado.

FVB/NJ strain as a mouse model for cutaneous leishmaniasis by Leishmania (L.) amazonensis

BACKGROUND Leishmaniases encompass a spectrum of neglected diseases caused by parasites of the genus Leishmania, grouped in two forms: tegumentary and visceral leishmaniasis. OBJECTIVES In this study, we propose Friend Virus B NIH Jackson (FVB/NJ) mouse strain as a new experimental model of infection with Leishmania (Leishmania) amazonensis, the second most prevalent agent of tegumentary leishmaniasis in Brazil. METHODS AND FINDINGS We performed in vitro infections of FVB/NJ macrophages and compared them with BALB/c macrophages, showing that BALB/c cells have higher infection percentages and a higher number of amastigotes/cell. Phagocytosis assays indicated that BALB/c and FVB/NJ macrophages have similar capacity to uptake parasites after 5 min incubations. We also investigated promastigotes' resistance to sera from FVB/NJ and BALB/c and observed no difference between the two sera, even though FVB/NJ has a deficiency in complement components. Finally, we subcutaneously infected FVB/NJ and BALB/c mice with 2 × 106 parasites expressing luciferase. Analysis of lesion development for 12 weeks showed that FVB/NJ and BALB/c mice have similar lesion profiles and parasite burdens. MAIN CONCLUSIONS This work characterises for the first time the FVB/NJ mouse as a new model for tegumentary leishmaniasis caused by Leishmania (L.) amazonensis.

Association of lipoprotein lipase (LPL) gene variants with hyperlipidemic acute pancreatitis in southeastern Chinese population

ABSTRACT Objective: The study aims to explore the relationship between lipoprotein lipase (LPL) variants and hyperlipidemic acute pancreatitis (HLAP) in the southeastern Chinese population. Subjects and methods: In total, 80 participants were involved in this study (54 patients with HLAP and 26 controls). All coding regions and intron-exon boundaries of the LPL gene were sequenced. The correlations between variants and phenotypes were also analysed. Results: The rate of rare LPL variants in the HLAP group is 14.81% (8 of 54), higher than in controls. Among the detected four variants (rs3735959, rs371282890, rs761886494 and rs761265900), the most common variant was rs371282890. Further analysis demonstrated that subjects with rs371282890 "GC" genotype had a 2.843-fold higher risk for HLAP (odds ratio [OR]: 2.843, 95% confidence interval [CI]: 1.119-7.225, p = 0.028) than subjects with the "CC" genotype. After adjusting for sex, the association remained significant (adjusted OR: 3.083, 95% CI: 1.208-7.869, p = 0.018). Subjects with rs371282890 "GC" genotype also exhibited significantly elevated total cholesterol, triglyceride and non-high-density lipoprotein cholesterol levels in all the participants and the HLAP group (p < 0.05). Conclusion: Detecting rare variants in LPL might be valuable for identifying higher-risk patients with HLAP and guiding future individualised therapeutic strategies.

O pequeno Édipo: a escuta de uma criança que "desconhece a sua origem" / Little Oedipus: listening to a child who "does not know his origin" / El pequeño Edipo: escuchando a un Niño que "No conoce su Origen" / Le petit Œdipe : l'écoute d'un enfant qui « ne connaît pas son origine ¼

Resumo Este estudo é um relato de experiência referente a um acompanhamento psicoterápico realizado com um menino de 9 anos, gerado por meio das tecnologias reprodutivas e que desconhece sua concepção. As discussões apresentadas emergiram durante as supervisões de estágio supervisionado em um serviço-escola de psicologia no período de maio a novembro de 2019. Durante as sessões, a criança trazia conteúdos a respeito de mitologia, natureza, filmes e jogos que, ao serem explorados em setting analítico, desvelaram o seu interesse em questões da sexualidade, da origem dos bebês e de sua própria concepção. Por meio deste relato, espera-se colaborar com reflexões sobre a omissão da concepção, em específico a reprodução assistida, e, ainda, possibilitar um aumento e uma atualização de estudos sobre a clínica psicanalítica com crianças no Brasil, de forma que venha a promover novas perspectivas de análise e contribuições.

Abstract This experience report refers to a psychotherapeutic counseling attended by a nine-year-old boy conceived through reproductive technologies and unaware of his conception. The discussions presented here emerged during a supervised internship at a Psychology School Service from May to November 2019. During the sessions, the child alluded to mythology, nature, movies and games contents which when explored in an analytical setting unveiled his interest in issues of sexuality, the origin of babies, and his own conception. We expect this report to collaborate with reflections on the omission of conception, especially in the context of assisted reproduction, and to increase and update child psychoanalytic research in Brazil, as to promote new perspectives of analysis.

Resumen Este estudio presenta un reporte de experiencia sobre un seguimiento psicoterapéutico de un niño de 9 años, generado mediante tecnologías reproductivas y que no conoce su concepción. Las discusiones presentadas surgieron durante las prácticas supervisadas en un Servicio Escolar de Psicología en el período de mayo a noviembre de 2019. Durante las sesiones, el niño trajo contenido sobre mitología, naturaleza, películas y juegos, que cuando se exploraba en un entorno analítico reveló su interés en temas de sexualidad, de origen de los bebés y de su propia concepción. Se espera mediante este reporte colaborar con las reflexiones sobre la omisión de la concepción, específicamente a la reproducción asistida, y también permitir un aumento y actualización de estudios de la clínica psicoanalítica infantil en Brasil, para que así se promuevan nuevas perspectivas de análisis y contribuciones.

Résumé Ce rapport d'expérience fait référence à un suivi psychothérapeutique réalisé avec un garçon de neuf ans conçu par des technologies de la reproduction et n'ayant pas conscience de sa conception. Les discussions présentées ici ont émergé lors d'un stage supervisé dans un service scolaire de psychologie de mai à novembre 2019. Au cours des séances, l'enfant a fait allusion à des informations sur la mythologie, la nature, les films et les jeux qui, lorsqu'il ont été explorés dans un cadre analytique, ont dévoilé son intérêt pour les questions de sexualité, l'origine des bébés et sa propre conception. Nous espérons que ce rapport collaborera avec les réflexions sur l'omission de la conception, en particulier dans le contexte de la procréation assistée, et qu'il augmentera et mettra à jour la recherche psychanalytique sur l'enfant au Brésil, afin de promouvoir de nouvelles perspectives d'analyse et de contributions.

Por que a psicologia do ego se distancia da obra freudiana? Uma análise do artigo de Ernst Kris em confronto com a obra de Freud à luz da leitura lacaniana / Why does ego psychology distance itself from the Freudian work? An analysis of Ernst Kris' article against Freud's work in light of the Lacanian reading / ¿Por qué la psicología del yo se distancia de la obra freudiana? Un análisis del artículo de Ernst Kris en comparación a la obra de Freud con base en la lectura lacaniana / Pourquoi la psychologie de l'ego se distancie-t-elle de l'œuvre freudienne ? Une analyse de l'article d'Ernst Kris en confrontation avec l'œuvre de Freud à la lumière de la lecture lacanienne

Resumo A psicologia do ego é reconhecida como uma releitura norte-americana da psicanálise, e teve Ernst Kris - que foi próximo de Freud - como um de seus fundadores. Apesar deste autor utilizar-se de textos freudianos para firmar os fundamentos de sua teoria, sustenta-se que a psicologia do ego mais se distancia do que se aproxima da obra de Freud. Este estudo visa demonstrar de que forma isso ocorre. Para isso, situa-se brevemente quem foi Ernst Kris e quais são as críticas já existentes à psicologia do ego. Num segundo momento, analisa-se o artigo "Ego Psychology and Interpretation in Psychoanalytic Therapy", de Kris, em confronto com a teoria de Freud. Por último, expõe-se a crítica de Lacan. Destaca-se que a crítica lacaniana remete a questões referentes à ética da psicanálise e ao lugar do analista na direção do tratamento.

Abstract Ego Psychology is a North-American re-reading of psychoanalysis and had Ernst Kris - a man who was close to Freud - as one of its founders. Despite using Freudian texts to establish its foundations, Ego Psychology distances itself from Freud's psychoanalysis. This study demonstrates how this occurs by briefly discussing who Ernst Kris was and the existing criticisms of Ego Psychology. Secondly, it analyzes Ernst Kris's article "Ego Psychology and Interpretation in Psychoanalytic Therapy" against Freud's theory, to finally present Lacan's critique, which refers to issues related to the ethics of psychoanalysis and the analyst's role in directing treatment.

Resumen La Psicología del Yo se considera como una relectura estadounidense del psicoanálisis y tuvo a Ernst Kris -quien era cercano a Freud- como uno de sus fundadores. A pesar de que Ernst Kris utiliza textos freudianos para establecer los fundamentos de su teoría, se sostiene que la Psicología del Yo se aleja más de la obra de Freud que se acerca a ella. Este estudio tiene como objetivo demostrar cómo ocurre esto. Para ello, se sitúa brevemente quién es Ernst Kris y cuáles son las críticas ya existentes a la Psicología del Yo. En segundo lugar, se analiza el artículo "Ego Psychology and interpretation in psychoanalytic therapy", de Ernst Kris, en confrontación con la teoría de Freud. Y, por último, se expone la crítica de Lacan. Se destaca que la crítica lacaniana remite a cuestiones relacionadas con la ética del psicoanálisis y el lugar del analista en la dirección del tratamiento.

Résumé La psychologie de l'égo est une relecture nord-américaine de la psychanalyse dont Ernst Kris, un homme proche de Freud, est l'un des fondateurs. Bien qu'elle utilise des textes freudiens pour établir ses fondements, la psychologie de l'égo s'éloigne de la pensée freudienne. Pour démontrer cet écart, cette étude aborde la figure de Ernst Kris et des critiques existantes à l'égard de la psychologie de l'égo. Ensuite, elle analyse l'article « Ego Psychology and interpretation in psychoanalytic therapy ¼ par rapport à la théorie de Freud, pour enfin présenter la critique de Lacan, qui se réfère à des questions liées à l'éthique de la psychanalyse et au rôle de l'analyste dans la direction du traitement.

Assessment of serum granulysin and cathepsin-L levels in vitiligo patients

SUMMARY OBJECTIVE: Cellular and humoral immunity plays a role in the pathogenesis of vitiligo. T lymphocytes and natural killer cells involved in cellular immunity carry out their cytotoxic activities through perforin/granzyme-dependent granule exocytosis, in which granulysin and cathepsin-L are also involved. The aim of this study was to investigate the possible role of serum granulysin and cathepsin-L in the etiopathogenesis of vitiligo and their association with disease activity and severity. METHODS: This randomized, prospective case-control study was conducted with 46 vitiligo patients admitted to the hospital for vitiligo between January and November 2021 and 46 healthy volunteers of similar age and gender. Serum levels of granulysin and cathepsin-L were measured by the enzyme-linked immunosorbent assay method. RESULTS: The mean serum levels of granulysin and cathepsin-L were statistically significantly higher in vitiligo patients compared with the control group (p=0.048 and p=0.024, respectively). There was no statistically significant correlation between serum granulysin and serum cathepsin-L levels and disease severity in the patient group (r=0.30, p=0.062 and r=0.268, p=0.071, respectively). Disease activity also showed no significant association with serum granulysin and cathepsin-L levels (p=0.986 and p=0.962, respectively). CONCLUSION: Although granulysin and cathepsin-L are molecules involved in the pathogenesis of vitiligo, the use of these molecules may not be helpful in assessing disease activity and severity. It may be helpful to conduct comprehensive and prospective studies to find new molecules to fill the gap in this area.

Problematização sobre o Transtorno Disruptivo da Desregulação do Humor / Problematization about Disruptive Mood Dysregulation Disorder / Problématisation du Trouble Disruptif avec Dysrégulation émotionnelle / Problematización del Trastorno de Desregulación Disruptiva del Estado de Ánimo

Este artigo se propõe a delinear, historicamente, o diagnóstico clínico do Transtorno Disruptivo da Desregulação do Humor (TDDH). Com base no método genealógico, essa categoria diagnóstica é desnaturalizada e recontextualizada em sua origem. Analisa-se o conceito de desregulação do humor a partir da escala CBCL (Childhood Behaviour Checklist), sua identificação como um transtorno bipolar da infância, posterior transformação no diagnóstico de TDDH e subsequente crítica deste, com a proposta de englobar os sintomas de desregulação do humor na infância, no diagnóstico de transtorno opositor-desafiante. Como alternativa, o artigo sugere que o humor irritadiço na infância é um estado afetivo primário, constituindo-se, assim, em uma predisposição orgânica primária. Já a regulação emocional é uma construção adaptativa, que se modela ao longo da vida, gerando apresentações subjetivas diversas.

Resumos This article historically outlines the clinical diagnosis of disruptive mood dysregulation disorder (DMDD). Based on the genealogical method, this diagnostic category is denaturalized and recontextualized in its origin. The concept of mood dysregulation is analyzed from the Childhood Behavior Checklist scale (CBCL), its identification as a childhood bipolar disorder, subsequent transformation in the diagnosis of DMDD and subsequent criticism of it, with the proposal of encompassing the symptoms of mood dysregulation humor in childhood into the diagnosis of oppositional defiant disorder. As an alternative, the article suggests that irritable mood in childhood is a primary affective state, thus, constituting a primary organic predisposition. Emotional regulation, on the other hand, is an adaptive construction, which is modeled throughout life, generating diverse subjective presentations.

Cet article retrace l'historique du diagnostic clinique du trouble disruptif avec dysrégulation émotionnelle (TDDE). Basée sur la méthode généalogique, cette catégorie diagnostique est dénaturalisée et recontextualisée dans ses origines. Le concept de dysrégulation émotionnelle est analysé à partir de l'échelle CBCL (Childhood Behavior Checklist), de son identification en tant que trouble bipolaire de l'enfance, de sa transformation ultérieure en diagnostic de TDDE et de sa critique ultérieure, avec la proposition d'inclure les symptômes de dysrégulation émotionnelle dans l'enfance dans le diagnostic du trouble oppositionnel avec provocation. Comme alternative, l'article suggère que l'humeur irritable dans l'enfance est un état affectif primaire, constituant ainsi une prédisposition organique primaire. La régulation émotionnelle, quant à elle, est une construction adaptative qui est modelée tout au long de la vie, générant diverses présentations subjectives.

Este artículo describe historicamente el diagnóstico clínico del trastorno de desregulación disruptiva del estado de ánimo (TDDEA). Con base en el método genealógico, esta categoria diagnostica se desnaturaliza y recontextualiza en su origen. Se analiza el concepto de desregulación del estado de ánimo a partir de la escala CBCL (Childhood Behavior Checklist), su identificación como trastorno bipolar pediátrico, posterior transformación en el diagnóstico de TDDEA y posterior crítica al mismo, con la propuesta de englobar los síntomas de desregulación del estado de ánimo en el diagnóstico de trastorno negativista desafiante. Como alternativa, se sugiere que el estado de ánimo de irritabilidad en la infancia es un estado afectivo primario, constituyendo una predisposición orgánica primaria. La regulación emocional, por su parte, es una construcción adaptativa, que se modela a lo largo de la vida, generando diversas presentaciones subjetivas.

Análise de metabólitos secundários presentes nas plantas Cynara scolymus L e Artemisia absinthium L durante o tratamento de dislipidemias / Analysis of secondary metabolites present in the plants Cynara scolymus L and Artemisia absinthium L during the treatment of dyslipidemia

O uso de plantas medicinais como forma de tratamento para várias doenças é uma tradição que se estende entre gerações, há milhares de anos. Os povos antigos costumavam usar partes específicas de alguns vegetais para a realização de chás e curativos para feridas. Ainda nos dias atuais, muitas pessoas preferem as formas naturais como tratamento para suas comorbidades, principalmente como adjuvantes no combate às dislipidemias. O presente trabalho teve o objetivo de analisar as plantas Cynara scolymus L e Artemisia absinthium L para confirmar a presença ou ausência de proteases específicas contra a hipercolesterolemia. A pesquisa foi realizada através de uma triagem das plantas no software RCSB (https://www.rcsb.org/). No final, pode-se confirmar que a planta Cynara scolymus L atende às perspectivas de tratamento da hipercolesterolemia, enquanto a Artemisia absinthium L não apresentou valores suficientes que sejam eficazes no tratamento.

The use of medicinal plants as a form of treatment for various diseases is a tradition that has spanned generations for thousands of years. Ancient people used to use specific parts of some vegetables to make teas and dressings for wounds. Even today, many people prefer natural forms as a treatment for their comorbidities, especially as adjuvants in the fight against dyslipidemia. The present work aimed to analyze the plants Cynara scolymus L and Artemisia absinthium L to confirm the presence or absence of specific proteases against hypercholesterolemia. The research was carried out by screening the plants using the RCSB software (https://www.rcsb.org/). In the end, it can be confirmed that the Cynara scolymus L plant meets the prospects for treating hypercholesterolemia, while Artemisia absinthium L did not present sufficient values that are effective in the treatment.

Falciparum malaria is associated with risk markers of type 2 diabetes mellitus in individuals with or without COVID-19 exposure

Background: Scientific information on the impact of malaria on the risk of developing type 2 diabetes mellitus (T2DM) after recovery from the coronavirus disease 2019 (COVID-19) is limited in the Ghanaian context. The purpose of this study was to examine the association between selected risk markers of T2DM in falciparum malaria patients post-COVID-19 or not at a tertiary hospital in Ghana. Methodology: This was a descriptive cross-sectional comparative study of 38-recovered COVID-19 adult participants with malaria and 40 unexposed COVID-19 adults with malaria at the Tamale Teaching Hospital, Ghana. Demographic, anthropometric and levels of glucose, insulin, C-reactive protein and lipid profiles were measured in the two groups of participants under fasting conditions. Parasitaemia was assessed microscopically but insulin resistance and beta-cell function were assessed by the homeostatic model. Results: The COVID-19 exposed participants were older (p=0.035) with lower parasitaemia (p=0.025) but higher mean levels of insulin, insulin resistance, and beta-cell function compared with their unexposed counterparts (p<0.05). Parasitaemia correlated positively with a number of the measured indices of diabetogenic risk markers in the COVID-19 exposed group only, and predicted (Adjusted R2=0.751; p=0.031) by beta-cell function, C-reactive protein and triglycerides with the model explaining about 75% of the observed variation. Parasitaemia could only be predicted (Adjusted R2=0.245; p=0.002) by C-reactive protein with the model explaining just about a quarter of the observed variation in the COVID-19 unexposed group. Insulin resistance and sub-optimal beta-cell function were detected in both groups of participants. Conclusion: Falciparum malaria is associated with risk markers for development of T2DM irrespective of COVID-19 exposure. Insulin resistance, inflammation and sub-optimal beta-cell secretory function may drive the risk. The observed diabetogenic risk is higher in the recovered COVID-19 participants.

Effect of microparticles derived from bone marrow mesenchymal stem cells on cardiomyocyte hypertrophy and its mechanism / 中国生物制品学杂志

@#Objective To investigate the effect of microparticles(MPs)derived from bone marrow mesenchymal stem cells(BMSCs) on myocardial hypertrophy and its mechanism.Methods The osteogenic differentiation and adipogenic differentiation of mesenchymal stem cells(MSCs) were induced. After isolation and purification,the morphological characteristics were observed by transmission electron microscope,and the MPs surface antigen was identified by flow cytometry. Myocardial hypertrophy model was induced by using isoprenaline(ISO)in rats,which were measured for the cardiac structure and function by echocardiography,and then detected for various indexes of the heart and isolated left ventricle. Single ventricular myocytes of rats were acutely isolated and divided into control group(Control group),cardiomyocyte hypertrophy group(ISO group),MPs group(MPs group),and MPs supernatant group(Supernatant group). The mRNA expressions of atrial natriuretic peptide(ANP)and B-type natriuretic peptide(BNP)were detected by qRTPCR. The expression levels of calmodulin-dependent protein kinaseⅡ（CaMKⅡ）and phosphorylated calmodulin-dependent protein kinaseⅡ（p-CaMKⅡ）were detected by ELISA. The L-type calcium current(LCa-L)in single ventricular myocyte of various groups was recorded by whole-cell patch clamp.Results The bone nodules of MSCs osteogenic differentiation turned red after alizarin red staining,and lipid droplets of adipogenic differentiation turned red after oil red O staining;Under transmission electron microscope,MPs membrane had a complete structure,a clear outline and a diameter of about200 nm;The positive rates of CD29 and CD90 on the surface of MPs were(98. 24 ± 0. 82)% and(97. 69 ± 1. 83)%,respectively. Compared with Control group,the left ventricular end diastolic dimension(LVEDD)reduced signifi-cantly(t =5. 065,P < 0. 05),while the interventricular septum end-diastolic dimension(IVSd),left ventricular posterior wall dimension(LVPWd),heart weight to body weight ratio(HW/BW),and heart weight to tibial length ratio(HW/Tibia)significantly increased in ISO group(t = 4. 013,2. 368,4. 392,5. 043 and 6. 120,respectively,each P < 0. 05),indicating that the hypertrophic model was successfully established. The expression levels of ANP and BNP mRNA in hypertrophic cardiomyocytes of rats in ISO group were significantly higher than those in Control group(t = 25. 120 and18. 261,respectively,each P < 0. 01);While the expression levels of ANP and BNP mRNA in MPs group significantly reduced after incubation with 48 μg/mL MPs for 48 h compared with ISO group(t = 12. 110 and 3. 526,respectively,each P < 0. 05);The expression levels of CaMK Ⅱand p-CaMKⅡ in ISO group were significantly higher than those in Control group(t = 3. 278 and 4. 181,respectively,each P < 0. 05),while the expression of p-CaMK Ⅱ in MPs group decreased significantly(t = 5. 420,P < 0. 05);The calcium current density in ISO group was significantly higher than that in Control group(t = 15. 261,P < 0. 01),while that in MPs group was significantly lower than that in ISO group(t =6. 216,P < 0. 05).Conclusion MSC-MPs can significantly inhibit ISO-induced cardiomyocyte hypertrophy in rats,which is related to its down-regulation of cardiomyocyte CaMKⅡ and inhibition of L-type calcium channel.

Construction and self-activation detection of yeast two-hybrid bait plasmid of human programmed cell death ligand 1 immunoglobulin variable region domain gene / 中国生物制品学杂志

@#Objective To construct a yeast two-hybrid recombinant bait plasmid of human programmed cell death ligand 1(PD-L1)immunoglobulin variable region(IgV)domain gene,detect its expression in yeast and detect the cytotoxicity and self-activation of PD-L1 IgV protein as well as the interaction between PD-L1 IgV and human thioredoxin(hTrx).Methods Human PD-L1 was analyzed by bioinformatics method,and primers were designed to amplify PD-L1 IgV domain based on the coding region of PD-L1 gene registered in NCBI GenBank database. PCR amplification was carried out with pENTERPD-L1 plasmid as template,and then cloned into yeast two-hybrid bait vector pGBKT7. The recombinant bait plasmid and pGBKT7 empty vector were transformed into Y2HGold yeast cells respectively,and the PD-L1 IgV gene and its expression were detected by PCR and Western blot;Meanwhile,the protein toxicity and self-activation of PD-L1 IgV were detected,and the interaction between PD-L1 IgV and hTrx was detected by drip plate method.Results The bioinformatics analysis results of PD-L1 were consistent with related reports. The recombinant bait plasmid pGBKT7-PD-L1 IgV was correctly constructed,and Y2HGold positive clone was obtained,in which PD-L1 IgV was stably expressed. The empty vector pGBKT7 and recombinant bait plasmid pGBKT7-PD-L1 IgV grew well on SD/-Trp and SD/-Trp/X-α-Gal plates with the same colony size and number and white colony,but they did not grow on SD/-Trp/X-α-Gal/AbA plates,which indicated that PD-L1 IgV protein had no toxicity and no self-activation effect on yeast. The results of drip plates test showed that all experimental groups grew well on SD/-Trp/-Leu plate,while only positive control group grew on SD/-Trp/-Leu/X-α-Gal/AbA plate and showed blue color,which indicated that bait protein PD-L1 IgV and hTrx did not self-activate,and there was no interaction between them.Conclusion Recombinant human PD-L1 IgV bait plasmid was successfully constructed. PD-L1 IgV protein showed no toxicity and self-activation effect on yeast cells,and there was no interaction between PD-L1 IgV and hTrx. Subsequently,hTrx can be used to construct a peptide aptamer library,from which peptide aptamers that specifically bind to PD-L1 IgV can be screened.

A new pyrazine from Hypecoum erectum L / 药学学报

Four pyrazines were isolated from the n-butanol fraction of Hypecoum erectum L. by using various chromatographic methods, including MCI gel, ODS, silica gel and semi-preparative HPLC. The structures of the isolated compounds were identified as hyperectpyrazin A (1), 1′S-(6-methylpyrazin-2-yl)-ethane-1′,2′-diol (2), 2-hydroxymethyl-6-methylpyrazin (3) and pyrazine-2-carboxylic acid (4) by spectroscopy methods (1D NMR, 2D NMR, UV, IR, MS, etc.). The absolute configuration of compound 2 was determined by using the Mo2(OAc)4 induced CD analysis for the first time. Compound 1 was a new compound, compounds 2-4 were isolated from H. erectum for the first time. Compounds 1-4 were evaluated for their inhibition against acetylcholinesterase and nitric oxide generation induced by lipopolysaccharide-RAW264.7 macrophage cells. At a concentration of 50 μmol·L-1, compounds 2 and 4 displayed inhibitory effects on acetylcholinesterase with the inhibition rates of 44.40% and 43.99%, respectively.

Preparation and in vitro evaluation of polylactic acid nanoparticles containing arginine and glucose oxidase / 药学学报

Hydrogen peroxide (H2O2) and nitric oxide (NO) has a short half-life, low bioavailability, poor tumor targeting and systemic adverse reactions in the physiological environment. In this study, phacoemulsification and nano-precipitation were used to synthesize didecyl dimethyl ammonium bromide (DDAB)/polylactic acid nanoparticles (PLA), then L-arginine (L-Arg) and glucose oxidase (GOx)-loaded nanoparticles (GADP) were prepared, and the in vitro antitumor activity was investigated．The particle size, potential, embedding rate and the ability to produce H2O2/NO of the nanoparticles were investigated. Meanwhile, in vitro cell cytotoxicity against human hepatoma cells (HepG2) was evaluated．The results showed that the prepared L-Arg-DDAB/PLA (ADP) nanoparticles were spherical particles. And the particle size and zeta potential were (225.7 ± 6.33) nm and (+23.5 ± 0.12) mV, respectively. The adsorption rate of GOx was 87.23% ± 0.02%. The drug loading of L-Arg was 15.6% ± 0.22%. The pH value of glucose solution and the amount of H2O2 showed that GADP had good catalytic activity. In vitro cytotoxicity experiments showed that blank nanoparticles were nontoxic, while the drug-loaded nanoparticles presented enhanced antitumor effect on HepG2 cells. And can inhibit tumor cell migration. The low dose nano-scale NO delivery system GADP can effectively inhibit the migration of tumor cells and kill tumor cells, thus producing therapeutic benefits.

Prevention and treatment of lipoic acid-niacin on blue-light-induced retinal damage in rats / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the preventive effect and optimal drug dose of lipoic acid-niacin(N2L)against blue light-induced retinal damage in SD rats, and to explore its possible protective mechanism.METHODS: A total of 36 specific pathogen free-grade male SD rats of 150-200 g were selected and randomly divided into normal control group, blue light injury group, N2L low-dose group(1.0 mg/kg), N2L medium-dose group(2.5 mg/kg), N2L high-dose group(5.0 mg/kg), and physiological saline group, with 6 rats in each group. The normal control group was reared in a 12 h dark and light cycle, and the rest of the groups received 9 h of daily light exposure, 3 h of blue light irradiation with a wavelength of 455 nm and an intensity of 3000±50 lx, and 12 h of darkness to establish the injury model, and were exposed to light exposure for 14 d. For 14 consecutive durations, a 1 mL dose of the corresponding drug was injected intraperitoneally. The rats were reared for another 5 d with a regular 12 h light-dark cycle and were examined by electroretinography. Specimens were prepared by over anesthesia, HE staining, and the thickness of the outer nuclear layer was observed under a optical microscope; superoxide dismutases(SOD)activity was detected by CheKineTM SOD Activity Assay Kit; and the retinal Caspase-3, quinone oxidoreductase 1(NQO1), glutathione S transferase(GST), Bcl-2, and Bax protein expression in rat retina were detected by Western blot.RESULTS: The amplitude of b-wave in dark-vision ERG 3.0 and 10.0(cd·s)/m2 stimulated light, b-wave in bright-vision ERG 3.0(cd·s)/m2 stimulated light, and the amplitude of the 2nd wave peak of oscillatory potential were significantly lower in blue light injury group than that in the normal control group(all P&#x0026;#x003C;0.01), while the amplitude was significantly higher in the N2L medium-dose group than in the blue light injury group(all P&#x0026;#x003C;0.05), and was not statistically different from that of the normal control group; the thickness of the retina in the blue light injury group was decreased in the ONL compared with that of the normal control group(P&#x0026;#x003C;0.001), while in the N2L medium dose group, it was thicker than that of the blue light injury group(P&#x0026;#x003C;0.001), and there was no statistically significant difference from the normal control group; SOD activity was significantly higher in the N2L medium-dose group than in the remaining 5 groups(P&#x0026;#x003C;0.05); the expression of Caspase-3, Bax, and NQO1 in the blue light injury group was higher than that of the normal control group(all P&#x0026;#x003C;0.01), and expression of Bax and Caspase-3 was significantly lower in the N2L medium-dose group compared with the blue light injury group(all P&#x0026;#x003C;0.001), whereas GST, NQO1 and Bcl-2 were significantly increased(all P&#x0026;#x003C;0.01).CONCLUSION:A concentration of 2.5 mg/kg N2L can effectively antagonize the damaging effect of blue light on the retina of SD rats, and it is expected to be a preventive and curative drug for it.

Anti-SARS-CoV-2 activity of small molecule inhibitors of cathepsin L / 药学学报

The coronavirus disease 2019 (COVID-19) is an acute infectious disease caused by the new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, which has led to serious worldwide economic burden. Due to the continuous emergence of variants, vaccines and monoclonal antibodies are only partial effective against infections caused by distinct strains of SARS-CoV-2. Therefore, it is still of great importance to call for the development of broad-spectrum and effective small molecule drugs to combat both current and future outbreaks triggered by SARS-CoV-2. Cathepsin L (CatL) cleaves the spike glycoprotein (S) of SARS-CoV-2, playing an indispensable role in enhancing virus entry into host cells. Therefore CatL is one of the ideal targets for the development of pan-coronavirus inhibitor-based drugs. In this study, a CatL enzyme inhibitor screening model was established based on fluorescein labeled substrate. Two CatL inhibitors IMB 6290 and IMB 8014 with low cytotoxicity were obtained through high-throughput screening, the half inhibition concentrations (IC50) of which were 11.53 ± 0.68 and 1.56 ± 1.10 μmol·L-1, respectively. SDS-PAGE and cell-cell fusion experiments confirmed that the compounds inhibited the hydrolysis of S protein by CatL in a concentration-dependent manner. Surface plasmon resonance (SPR) detection showed that both compounds exhibited moderate binding affinity with CatL. Molecular docking revealed the binding mode between the compound and the CatL active pocket. The pseudovirus experiment further confirmed the inhibitory effects of IMB 8014 on the S protein mediated entry process. In vitro pharmacokinetic evaluation indicated that the compounds had relatively good drug-likeness properties. Our research suggested that these two compounds have the potential to be further developed as antiviral drugs for COVID-19 treatment.

Enhancement of tropane alkaloids biosynthesis in Atropa belladonna hariy root by overexpression of HnCYP82M3 and DsTRI genes / 药学学报

Tropane alkaloids (TAs) are a class of anticholinergic drugs widely used in clinical practice and mainly extracted from plant, among which Atopa belladonna is the main commercial drug source. It is of great industrial value to obtain TAs in large quantities by plant metabolic engineering. In TAs pathway, cytochrome oxidase CYP82M3 catalyze the synthesis of tropinone and then tropinone reductase I (TRI) compete with TRII for tropinone to form tropine leading to the TAs synthesis (drainage). In this study, based on the "increasing flow and drainage" metabolic engineering strategy, two genes, namely HnCYP82M3 and DsTRI from Hyoscyamus niger and Datura stramonium, respectively, were overexpressed in the hair roots of A. belladonna, with a view to promote the TAs accumulation. The HnCYP82M3 gene was cloned from the root of H. niger, and it encoded amino acid with 91.7% sequence identity with AbCYP82M3 from A. belladonna. Overexpression of HnCYP82M3 alone did not affect the content of TAs in hair roots of A. belladonna, indicating that CYP82M3 was not a key enzyme in TAs biosynthesis. Simultaneous overexpression of HnCYP82M3 and DsTRI greatly promoted the accumulation of the three TAs, and the contents of hyoscyamine, anisodamine and scopolamine were 4.97 times, 2.83 times and 2.19 times that of the control, respectively, and the increase amplitude was greater than that of single overexpression of DsTRI. This study showed that the "increasing flow and drainage" strategy of enzyme genes co-expression at branch points was a promising metabolic engineering method to effectively improve the biosynthesis of TAs in A. belladonna, and laid a theoretical and technical foundation for the large-scale industrial acquisition of TAs.

Blumea balsamifera （L.） DC Oil Alleviates LPS-Induced Inflammatory Response in Macrophages by Affecting Arachidonic Acid Metabolism Via NF-κB Nonclassical Pathway / 中山大学学报(医学科学版)

ObjectiveTo study the anti-inflammatory effects of Blumea balsamifera （L.） DC oil （BBO） based on nuclear factor kappa-B （NF-κB） nonclassical and arachidonic acid （AA） pathway. MethodsEffects of BBO on the production of slow reacting substance of anaphylaxis （SRS-A） were detected by the ileal smooth muscle method. The contents of prostaglandin E2 （PGE2） and leukotriene B4 （LTB4） in lipopolysaccharides （LPS） -induced macrophages were detected by ELISA kit. The expression of COX-2， 5-LOX， FLAP and RelB were detected by qRT-PCR. Western blot was performed to detect the effects of BBO on the level of NF-κB nonclassical pathway proteins TNF receptor associated factor 3 （TRAF3）， TNF receptor associated factor 2 （TRAF2）， NF-κB-inducing kinase （NIK）， p100 and RelB. ResultsThe contractile tension of guinea pig ileum was reduced （P<0.001）， and the SRS-A production inhibition rate reached 65.34% at 1mg·mL-1 BBO concentration. Compared with LPS group， BBO reduced the concentrations of PGE2 （P<0.05） and LTB4 （P<0.05）， and decreased the expressions of COX-2 （P<0.05）， 5-LOX （P<0.05） and FLAP （P<0.05） in AA pathway at concentrations of 40-80 μg·mL-1. Moreover， 40-80 μg·mL-1 BBO decreased the concentrations of TRAF3 （P<0.05）， TRAF2 （P<0.05）， and NIK （P<0.05）， and further inhibited the phosphorylation of p100 （P<0.05）， as well as the level of the transcription factor RelB in genes （P<0.05） and proteins （P<0.05） in nonclassical NF-κB pathway， whereas BBO did not cause such changes. ConclusionBBO may potentially exert its anti-inflammatory effects by suppressing the regulatory proteins TRAF3 and TRAF2 and the transcription factor RelB in NF-κB nonclassical pathway. The inhibitory action extending to the induction kinase function of NIK， further hindering the phosphorylation of p100 and its binding with the transcription factor RelB. Consequently， downstream elements in the AA pathway， including the pivotal rate-limiting enzymes COX-2， 5-LOX and FLAP， were altered. This modulation influences the levels of inflammatory mediators such as PGE2 and LTB4.

Clinical significance of determining the level of biliary calprotectin in patients with cholangiocarcinoma or choledocholithiasis / 临床肝胆病杂志

ObjectiveTo investigate the difference in the level of biliary calprotectin between patients with cholangiocarcinoma and those with choledocholithiasis. MethodsClinical data and bile samples were collected from 34 patients with cholangiocarcinoma and 78 patients with choledocholithiasis who were diagnosed and treated with endoscopic retrograde cholangiopancreatography in The First Affiliated Hospital of Anhui Medical University from May 2021 to September 2022. Fluorescence lateral flow immunoassay was used to measure the levels of calprotectin， hemoglobin， and lactoferrin in bile. The Mann-Whitney U test was used for comparison of continuous data between two groups， and the chi-square test was used for comparison of categorical data between two groups； the Spearman correlation test was used for correlation analysis； the DeLong test was used for comparison of the area under the ROC curve （AUC）. ResultsCompared with the choledocholithiasis group， the cholangiocarcinoma group had significant increases in the levels of calprotectin ［4 795.50 （2 286.79‍ ‍—‍ ‍20 179.73） ng/mL vs 411.16 （67.03‍ ‍—‍ ‍1 991.88） ng/mL， Z=5.572， P<0.001］ and fluoride ［115.70 （109.10‍ — ‍125.50） mmol/L vs 106.60 （98.60‍ ‍—‍ ‍114.40） mmol/L， Z=2.702， P=0.007］. The patients with cholangiocarcinoma were further divided into high cholangiocarcinoma group and low cholangiocarcinoma group， and there was no significant difference between the two groups in the level of calprotectin ［3 867.71 （2 235.66‍ — ‍26 407.40） ng/mL vs 4 795.50 （2 361.15‍ — ‍13 070.53） ng/mL， Z=0.129， P>0.05］. Biliary calprotectin level was correlated with white blood cell count， hemoglobin concentration， and lactoferrin concentration in bile （r=0.316， 0.353， and 0.464， all P<0.05）. The ROC curve analysis showed that biliary calprotectin （with a sensitivity of 79.4% and a specificity of 75.6%）， blood CA19-9 （with a sensitivity of 82.4% and a specificity of 78.2%）， and their combination （with a sensitivity of 88.2% and a specificity of 73.1%） had good sensitivity and specificity in the diagnosis of cholangiocarcinoma. ConclusionThere is an increase in the level of biliary calprotectin in patients with cholangiocarcinoma， and therefore， it might become a biomarker for the diagnosis of cholangiocarcinoma.

Cancer immunotherapy with enveloped self-amplifying mRNA CARG-2020 that modulates IL-12, IL-17 and PD-L1 pathways to prevent tumor recurrence

Targeting multiple immune mechanisms may overcome therapy resistance and further improve cancer immunotherapy for humans. Here, we describe the application of virus-like vesicles (VLV) for delivery of three immunomodulators alone and in combination, as a promising approach for cancer immunotherapy. VLV vectors were designed to deliver single chain interleukin (IL)-12, short-hairpin RNA (shRNA) targeting programmed death ligand 1 (PD-L1), and a dominant-negative form of IL-17 receptor A (dn-IL17RA) as a single payload or as a combination payload. Intralesional delivery of the VLV vector expressing IL-12 alone, as well as the trivalent vector (designated CARG-2020) eradicated large established tumors. However, only CARG-2020 prevented tumor recurrence and provided long-term survival benefit to the tumor-bearing mice, indicating a benefit of the combined immunomodulation. The abscopal effects of CARG-2020 on the non-injected contralateral tumors, as well as protection from the tumor cell re-challenge, suggest immune-mediated mechanism of protection and establishment of immunological memory. Mechanistically, CARG-2020 potently activates Th1 immune mechanisms and inhibits expression of genes related to T cell exhaustion and cancer-promoting inflammation. The ability of CARG-2020 to prevent tumor recurrence and to provide survival benefit makes it a promising candidate for its development for human cancer immunotherapy.