In vivo studies could not confirm in vitro prophylactic synergism between Moringa essential oil and Lactobacillus reuteri (MT180537) / Estudos in vivo não puderam confirmar o sinergismo profilático in vitro entre o óleo essencial de Moringa e Lactobacillus reuteri (MT180537)

Aerobic vaginitis (AV) is a recently defined vaginal recurring infection, which is treated with antibiotics. However, excessive and prolonged use of antibiotics disrupts healthy vaginal microflora and leads to the emergence of antibiotic resistance among pathogens. This situation has directed researchers to explore alternative antimicrobials. The current study describes in vitro and in vivo antimicrobial efficacy and pharmaceutical interactions between plant essential oils (EOs) and five lactic acid bacteria (LABs), isolated from the healthy vagina, against E. faecalis, one of the major etiological agents of AV. In vitro experiments confirm good antimicrobial activity of both plant EOs and cell free supernatant (CFS) from LABs. Based on high antimicrobial efficacy, Moringa essential oil (MO) was selected to determine its nature of interaction with CFS of five LAB strains. Synergism was recorded between MO and CFS of L. reuteri (MT180537). To validate in vitro findings, prophylactic responses of individual and synergistic application of MO and L. reuteri (MT180537) were evaluated in an E. faecalis (MW051601) induced AV murine model. The prophylactic efficacy was evidenced by a reduction in intensity of clinical symptoms, E. faecalis (MW051601) count per vaginal tissue along with a reduction in AV associated changes in histological markers of infection in animals receiving Moringa essential oil and L. reuteri (MT180537) alone or in combination. However, significant synergism between Moringa essential oil and L. reuteri (MT180537) could not be observed. Our data confirms the importance of in vivo experiments in deducing pharmacological interactions.

Vaginite aeróbica (VA) é uma infecção vaginal recorrente definida recentemente, que é tratada com antibióticos. No entanto, o uso excessivo e prolongado de antibióticos perturba a microflora vaginal saudável e leva ao surgimento de resistência aos antibióticos entre os patógenos. Esta situação levou os pesquisadores a explorar antimicrobianos alternativos. O presente estudo descreve a eficácia antimicrobiana in vitro e in vivo e as interações farmacêuticas entre óleos essenciais vegetais (OE) e cinco bactérias lácticas (BAL), isoladas de vagina sã, contra E. faecalis, um dos principais agentes etiológicos da AV. Os experimentos in vitro confirmam a boa atividade antimicrobiana de ambos os EOs de plantas e sobrenadante livre de células (CFS) de LABs. Com base na alta eficácia antimicrobiana, o óleo essencial de Moringa (MO) foi selecionado para determinar sua natureza de interação com o sobrenadante livre de células (CFS) de cinco cepas de LAB. Sinergismo foi registrado entre MO e CFS de L. reuteri (MT180537). Para validar os resultados in vitro, as respostas profiláticas da aplicação individual e sinérgica de MO e L. reuteri (MT180537) foram avaliadas em um modelo murino AV induzido por E. faecalis (MW051601). A eficácia profilática foi evidenciada por uma redução na intensidade dos sintomas clínicos, contagem de E. faecalis (MW051601) por tecido vaginal, juntamente com uma redução nas alterações associadas a AV nos marcadores histológicos de infecção em animais que receberam óleo essencial de Moringa e L. reuteri (MT180537) sozinho ou em combinação. No entanto, não foi possível observar sinergismo significativo entre o óleo essencial de Moringa e L. reuteri (MT180537). Nossos dados confirmam a importância dos experimentos in vivo na dedução de interações farmacológicas.

Preparation,identification and comparison of murine and rabbit polyclonal antibodies against matrix protein of rabies virus strain CVS-11 / 中国生物制品学杂志

@#Objective To prepare murine and rabbit polyclonal antibodies against rabies virus(RV) matrix(M) protein and compare their reactivity.Methods The prokaryotic expression vector pET-28a-M was constructed by using the cDNA of cells infected with RV CVS-11 strain as template,then transformed into E.coli BL21(DE3),and the induced by IPTG to express M protein.After nickel column affinity chromatography and dialysis renaturation,female BALB/c mice and New Zealand white rabbits were immunized with the M protein,and the whole blood was taken to separate the serum.The titers of the murine and rabbit polyclonal antibodies were detected by ELISA,and the reactivity was measured by Western blot,indirect immunofluorescence assay(IFA) and immunoprecipitation(IP).Results The plasmid pET-28a-M was constructed correctly as identified by sequencing.The titers of murine and rabbit polyclonal antibodies were 1:100 and 1:256 000respectively,and the polyclonal antibodies had reactivity with different RV strains.Conclusion The murine and rabbit polyclonal antibodies against M protein were successfully prepared,which provides important biological tools for exploring the interaction between M protein and host protein as well as studying the pathogenesis of RV.

In vivo studies could not confirm in vitro prophylactic synergism between Moringa essential oil and Lactobacillus reuteri (MT180537)

Abstract Aerobic vaginitis (AV) is a recently defined vaginal recurring infection, which is treated with antibiotics. However, excessive and prolonged use of antibiotics disrupts healthy vaginal microflora and leads to the emergence of antibiotic resistance among pathogens. This situation has directed researchers to explore alternative antimicrobials. The current study describes in vitro and in vivo antimicrobial efficacy and pharmaceutical interactions between plant essential oils (EOs) and five lactic acid bacteria (LABs), isolated from the healthy vagina, against E. faecalis, one of the major etiological agents of AV. In vitro experiments confirm good antimicrobial activity of both plant EOs and cell free supernatant (CFS) from LABs. Based on high antimicrobial efficacy, Moringa essential oil (MO) was selected to determine its nature of interaction with CFS of five LAB strains. Synergism was recorded between MO and CFS of L. reuteri (MT180537). To validate in vitro findings, prophylactic responses of individual and synergistic application of MO and L. reuteri (MT180537) were evaluated in an E. faecalis (MW051601) induced AV murine model. The prophylactic efficacy was evidenced by a reduction in intensity of clinical symptoms, E. faecalis (MW051601) count per vaginal tissue along with a reduction in AV associated changes in histological markers of infection in animals receiving Moringa essential oil and L. reuteri (MT180537) alone or in combination. However, significant synergism between Moringa essential oil and L. reuteri (MT180537) could not be observed. Our data confirms the importance of in vivo experiments in deducing pharmacological interactions.

Resumo Vaginite aeróbica (VA) é uma infecção vaginal recorrente definida recentemente, que é tratada com antibióticos. No entanto, o uso excessivo e prolongado de antibióticos perturba a microflora vaginal saudável e leva ao surgimento de resistência aos antibióticos entre os patógenos. Esta situação levou os pesquisadores a explorar antimicrobianos alternativos. O presente estudo descreve a eficácia antimicrobiana in vitro e in vivo e as interações farmacêuticas entre óleos essenciais vegetais (OE) e cinco bactérias lácticas (BAL), isoladas de vagina sã, contra E. faecalis, um dos principais agentes etiológicos da AV. Os experimentos in vitro confirmam a boa atividade antimicrobiana de ambos os EOs de plantas e sobrenadante livre de células (CFS) de LABs. Com base na alta eficácia antimicrobiana, o óleo essencial de Moringa (MO) foi selecionado para determinar sua natureza de interação com o sobrenadante livre de células (CFS) de cinco cepas de LAB. Sinergismo foi registrado entre MO e CFS de L. reuteri (MT180537). Para validar os resultados in vitro, as respostas profiláticas da aplicação individual e sinérgica de MO e L. reuteri (MT180537) foram avaliadas em um modelo murino AV induzido por E. faecalis (MW051601). A eficácia profilática foi evidenciada por uma redução na intensidade dos sintomas clínicos, contagem de E. faecalis (MW051601) por tecido vaginal, juntamente com uma redução nas alterações associadas a AV nos marcadores histológicos de infecção em animais que receberam óleo essencial de Moringa e L. reuteri (MT180537) sozinho ou em combinação. No entanto, não foi possível observar sinergismo significativo entre o óleo essencial de Moringa e L. reuteri (MT180537). Nossos dados confirmam a importância dos experimentos in vivo na dedução de interações farmacológicas.

Research Progress of BRAF Fusion in Non-small Cell Lung Cancer / 中国肺癌杂志

In advanced non-small cell lung cancer (NSCLC), V-Raf murine sarcoma viral oncogene homolog B1 (BRAF) mutation is highly malignant and has poor prognosis, and currently Dabrafenib in combination with Trametinib is approved for first-line treatment of patients with BRAF V600 mutation. In addition to mutations, BRAF fusion can also occur. With the development of gene detection, the detection of BRAF fusion is gradually increasing, but there is a lack of effective therapeutic strategies for BRAF fusion. In this paper, we review the clinical characteristics, mechanism of action, and clinical treatment of BRAF fusion to provide a basis for the treatment of BRAF fusion in NSCLC patients.
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Development and validation of real-time fluorescence quantitative PCR method for murine virus / 中国生物制品学杂志

@#Objective To develop and validate the real-time fluorescent quantitative PCR(Q-PCR)method for the detection of 8 murine viruses. Methods The specificity,sensitivity and precision of the Q-PCR method were verified by four laboratories,and the virus simulated contamination test and blind sample detection were carried out simultaneously,of which the detection results were compared. The Q-PCR method was used to detect 26 batches of monoclonal antibody cell lines for SARS-CoV-2 vaccine production and 15 batches of other products of murine origin. Results The Q-PCR method used for detecting 8 kinds of murine viruses had no cross reaction with the same family and genus or other murine viruses. Except the sensitivity of laboratory 2 to ectromelia virus(EctV/Mouse Pox,MPV)was 2 × 10~2copies/μL,the sensitivity of laboratory 2 to other 7 viruses and 3 other laboratories to 8 murine viruses was 2 × 10~1copies/μL. Except the inter-assay CV of the copy number of mouse adenovirus(MAdV)detected by laboratory 3 was 37. 58%,the intra-assay and inter-assay CVs of the Ct and copy number of other 7 viruses detected by laboratory 3 and those of 8 viruses detected by other 3 laboratories were all less than 25%.The sensitivity of virus simulated contamination test met the parameter requirements. The coincidence rate of blind sample detection results by 4 laboratories was 100%. All the 26 batches of monoclonal antibody cell lines for SARS-CoV-2 vaccine production and 15 batches of other murine derived products were negative for 8 murine viruses. Conclusion Q-PCR method for murine virus has good specificity,sensitivity and precision,and can be used for the detection of murine derived biological products.

Exploration of hapten-induced atopic dermatitis murine models for non-clinical pharmacodynamics study of drugs / 药学学报

Atopic dermatitis (AD) is a chronic, relapsing, inflammatory dermatosis with a variety of clinical manifestations and difficult to cure. Currently, many AD drug candidates have entered the research and development pipeline. In order to provide technical specifications for the clinical development of AD drugs, the Center for Drug Evaluation of National Medical Products Administration released the "Technical Guidelines for Clinical Trials of Drugs for AD Treatment" (Draft for Comments) in November 2022. Non-clinical pharmacodynamics evaluation is an important research before the drug enters clinical trials. Oxazolone (OXA)- and 2,4-dinitro-fluorobenzene (DNFB)-induced models are the most popular classical hapten-induced AD murine models, but variations of modeling are existing in the methods from different studies, including sensitization sites, haptens' dosages, the period of challenges, and the skin lesions severity evaluation as well. In this study, the investigation of OXA- and DNFB-induced AD murine models with various conditions of modeling was performed to compare the characteristics of hapten-induced AD murine models in the pathological process and severity according to the appearance of AD patients, and the guidance of pharmacodynamics evaluation of AD-therapeutic drugs in clinical trials as well, which may provide a proposal for AD treatment drug candidates in the non-clinical pharmacodynamics evaluation. All animal experiments were approved by the Animal Care & Welfare Committee of Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College (approval No.: 00007782 and 00007784).

Effect of aluminum exposure on abnormal phosphorylationof tau protein in PC12 cells of rats / 预防医学

Objective @#To investigate the effect of aluminum exposure on expression of miR-497-5p, wingless murine breast cancer virus integration site family member 3a (Wnt3a), β-catenin protein, glycogen synthase kinase-3β (GSK-3β) protein and tau protein in rat adrenal pheochromocytoma PC12 cells, so as to provide insight into unraveling the mechanisms underlying aluminum exposure-induced abnormal phosphorylation of tau protein.@* Methods@# PC12 cells were exposed to Al(mal)3 at concentrations of 0, 100, 200, 400 μmol/L for 24 h. The viability of PC12 cells was measured using cell counting kit-8 (CCK-8) assay. The relative expression of miR-497-5p and Wnt3a was detected using a real-time fluorescent quantitative PCR (RT-qPCR) assay, and the expression of Wnt3a, β-catenin, GSK-3β, P-GSK-3β (Ser9), tau and p-tau (Ser396) proteins were determined using Western blotting. @*Results @#The viability of PC12 cells appeared a tendency towards a decline with the increase of aluminum dose (Ftrend=323.473, P=0.001). RT-qPCR assay detected that the relative miR-497-5p expression appeared a tendency towards a rise with the increase of aluminum dose (Ftrend=14.888, P=0.031), and the relative Wnt3a expression appeared a tendency towards a decline with the increase of aluminum dose (Ftrend=165.934, P<0.001). The miR-497-5p expression negatively correlated with the relative Wnt3a expression (r=-0.693, P=0.012). The expression of Wnt3a (Ftrend=357.656, P=0.001), β-catenin (Ftrend=208.750, P=0.001) and p-GSK-3β (Ser9) proteins (Ftrend=512.583, P<0.001) appeared a tendency towards a decline with the increase of aluminum dose, and the expression of GSK-3β (Ftrend=39.965, P<0.001), tau (Ftrend=277.929, P=0.006) and p-tau (Ser396) proteins (Ftrend=96.247, P=0.002) appeared a tendency towards a rise with the increase of aluminum dose. @*Conclusion@# Up-regulation of miR-497-5p and GSK-3β expression and down-regulation of Wnt3a and β-catenin expression may be a mechanism underlying aluminum exposure-induced abnormal phosphorylation of tau protein.

Salmonella entérica serovar Enteritidis biofilm lifestyle induces lower pathogenicity and reduces inflammatory response in a murine model compared to planktonic bacteria / Salmonella entérica serovar Enteritidis en biopelícula induce una menor patogenicidad y respuesta inflamatoria en un modelo murino que su contraparte planctónica

Abstract Salmonella enterica serovar Enteritidis (S. Enteritidis) is the most frequent serovar involved in human salmonellosis. It has been demonstrated that about 80% of infections are related to biofilm formation. There is scant information about the pathogenicity of S. Enteritidis and its relationship to biofilm production. In this regard, this study aimed to investigate the differential host response induced by S. Enteritidis biofilm and planktonic lifestyle. To this purpose, biofilm and planktonic bacteria were inoculated to BALB/c mice and epithelial cell culture. Survival studies revealed that biofilm is less virulent than planktonic cells. Reduced signs of intestinal inflammation and lower bacterial translocation were observed in animals inoculated with Salmonella biofilm compared to the planktonic group. Results showed that Salmonella biofilm was impaired for invasion of non-phagocytic cells and induces a lower inflammatory response in vivo and in vitro compared to that of planktonic bacteria. Taken together, the outcome of Salmonella-host interaction varies depending on the bacterial lifestyle.

Resumen Salmonella entérica serovar Enteritidis (S. Enteritidis) es la serovariedad más frecuentemente aislada en la salmonelosis humana. Se ha demostrado que alrededor del 80% de las infecciones están relacionadas con la formación de biopelículas. Sin embargo, la información disponible acerca de la patogenicidad de S. Enteritidis y su relación con la producción de biopelículas es escasa. Este trabajo tuvo como objetivo investigar la respuesta diferencial del huésped frente a S. Enteritidis en sus 2 estilos de vida: biopelícula y planctónico. Para ello, se inocularon bacterias en estado de biopelícula o planctónico en ratones BALB/c y cultivo de células epiteliales. Los estudios de supervivencia revelaron que Salmonella en biopelícula fue menos virulenta que su contraparte planctónica. Los animales inoculados con biopelículas presentaron una mayor conservación estructural del intestino y una menor translocación bacteriana que el grupo planctónico. Asimismo, Salmonella en biopelícula mostró una capacidad deficiente para invadir células no fagocíticas e indujo una menor respuesta inflamatoria in vivo e in vitro que las bacterias planctónicas. Se concluye que el resultado de la interacción Salmonella-huésped depende del estilo de vida bacteriano.

Prevalence of rickettsias in ticks (Acari: Ixodidae) collected in domestic animals of the northern region of Caldas department, Colombia / Prevalencia de rickettsias en garrapatas (Acari: Ixodidae) colectadas en animales domésticos del norte del departamento de Caldas, Colombia

ABSTRACT The northern region of the Caldas department in Colombia is considered an endemic area for murine typhus. Recent studies in patients with acute febrile disease demonstrated infection with the spotted fever group's rickettsiae due to an increase in the IgG titer by indirect immunofluorescence in paired sera obtained from these patients. The objective of the current research was to identify the species of ticks present in domestic animals in the northern region of Caldas and establish the presence of rickettsial genomic material in the collected ticks. Ticks were obtained from bovines, horses, and dogs in seven municipalities in the north of Caldas. Ticks were stored in 90 % ethanol until processing and were identified using taxonomic keys, DNA was extracted using commercial techniques, and the gltA gene was amplified by conventional chain reaction polymerase (PCR). Seven hundred thirteen ticks were obtained from 593 domestic animals. The highest infestation occurred in cattle, followed by canines and horses. Ticks found corresponded to the species Riphicephalus microplus, Dermacentor nitens, Amblyomma sp., and Riphicephalus sanguineus s.l. In none of the tick samples, Rickettsia-specific gltA gene DNA was found. It can be inferred that the ticks obtained are not a source of rickettsial infection for people in this department region, despite finding different species associated with the transmission of this disease.

RESUMEN La región norte del departamento de Caldas, Colombia es considerada como una zona endémica de tifo murino. Estudios recientes realizados en pacientes con enfermedad febril aguda, demostraron la infección con rickettsias, del grupo de las fiebres manchadas, debido al aumento en el título de IgG, por inmunofluorescencia indirecta (IFI), en sueros pareados, obtenidos de dichos pacientes. El objetivo de la investigación fue el de identificar las especies de garrapatas presentes en animales domésticos, de la región norte de Caldas y establecer la presencia de material genómico de rickettsias, en las garrapatas recolectadas. En siete municipios, se recolectaron garrapatas de bovinos, de equinos y de caninos. Las garrapatas, se almacenaron en etanol al 90 %, hasta su identificación taxonómica. Se extrajo el ADN, utilizando técnicas comerciales y se amplificó por reacción de cadena de la polimerasa (PCR) convencional el gen gltA. Se obtuvieron 713 garrapatas de 593 animales domésticos. La más alta infestación se presentó en bovinos, seguido de los caninos y equinos. Las garrapatas encontradas correspondieron a las especies Riphicephalus microplus, Dermacentor nitens, Amblyomma sp. y Riphicephalus sanguineus s.l. En ninguna de las muestras, se comprobó la presencia de ADN del gen gltA específico de Rickettsia. Se puede inferir que las garrapatas obtenidas no serían una fuente de infección rickettsial para las personas, en esta región del departamento; sin embargo, su presencia es un factor de riesgo para la adquisición de rickettsiosis asociadas con las fiebres manchadas.

Hepatitis E virus infection and gene polymorphism in murine-shaped animals of plague foci in Yunnan Province / 中华地方病学杂志

Objective:To learn about the infection and gene polymorphisms of hepatitis E virus (HEV) in murine-shaped animals of plague foci in Yunnan Province.Methods:From July to August 2019, around 16 natural villages (4 in Mile City, 6 in Mangshi and 6 in Lianghe County), which were the foci of domestic plague in Yunnan Province, the murine-shaped animals were captured by the night-time method. The liver RNA was extracted, and the target gene of rat HEV was detected by one-step real-time fluorescence quantitative PCR, and the positive rate of rat HEV was calculated. The rat HEV positive samples were amplified by PCR for further clone sequencing, and the resulting sequences were compared with the HEV sequences registered in the GenBank and a phylogenetic tree was constructed by using MegAlign and MEGA 7.0.Results:A total of 491 murine-shaped animals were captured from 3 orders, 5 families, 8 genera, 15 species, and the positive rate of rat HEV was 4.89% (24/491). Among them, the positive rate of Rattus tanezumi and Niviventer fulvescens was 9.39% (23/245) and 1/3, respectively; and other species were negative. There was a statistically significant difference in the positive rate of rat HEV between different habitats ( P = 0.014), and the positive rate of rat HEV in the habitats near the dwellings was higher than that in other habitats ( P < 0.05). The sequence comparison analysis showed that the gene sequence of P018 from Lianghe County was 100.0% homologous to the MG813927.1 sequence of the first patient with rat HEV in Hongkong, and it was clustered into the same branch with the sequences of MG813927.1 and LC549185.1 from rat, was the type HEV C. G024 from Mangshi shared a low homology (20.7% - 31.5%) with other virus strains, and it was clustered into the same branch with a HEV sequence from an avian (AY535004.1). Conclusions:Rat HEV is prevalent in murine-shaped animals of plague foci of Yunnan Province, and there may be gene polymorphisms of rat HEV. In addition, the difference infection rate may be related to the habitats.

Efecto antiinflamatorio y antioxidante in vitro de Curcuma longa sobre macrófagos murinos durante la infección por Helicobacter pylori

RESUMEN Introducción: La colonización por Helicobacter pylori produce una inflamación en la mucosa gástrica con el consecuente desarrollo de enfermedades gastroduodenales. Frente a altas tasas de resistencia antimicrobiana y la ausencia de una vacuna en humanos, la alternativa ha sido la búsqueda de extractos de plantas con propiedades antimicrobiana, antiinflamatoria, antioxidante, antifúngica y anticancerígena como la Curcuma longa. Sin embargo, al ser una especie introducida y adaptada a las condiciones climáticas del país, son necesarios los estudios preclínicos que avalen su potencial antiinflamatorio y antioxidante. Objetivo: Evaluar el efecto del extracto de C. longa sobre macrófagos peritoneales infectados con H. pylori. Métodos: Para evaluar el efecto antiinflamatorio y antioxidante del extracto de C. longa sobre macrófagos murinos infectados por H. pylori, se evaluaron diferentes concentraciones del extracto y relaciones de bacteria, y se evaluó la muerte celular mediante DAPI. Se determinó la producción de óxido nítrico, peróxido de hidrógeno y los niveles de la interleucina-1β. Resultados: La viabilidad del macrófago se afectó frente a concentraciones de 100 µg/mL del extracto de cúrcuma y a partir de 25 bacterias/macrófago. Al combinar las diferentes concentraciones del extracto con las multiplicidades bacterianas se observó una reducción en los niveles de H2O2 e IL-1β; sin embargo, la reducción del óxido nítrico se observó en el rango de 6,25-50 µg/mL del producto natural. Conclusiones: El extracto de cúrcuma cubano mostró potencial antioxidante y antiinflamatorio al disminuir la citotoxicidad celular y la producción de especies reactivas del oxígeno en macrófagos peritoneales.

ABSTRACT Introduction: Colonization by Helicobacter pylori causes inflammation of the gastric mucosa with the consequent development of gastroduodenal diseases. In view of the high antimicrobial resistance rates and the absence of a vaccine for humans, the alternative has been to search for plant extracts with antimicrobial, anti-inflammatory, antioxidant, antifungal and anticancer properties. An example is Curcuma longa. However, being as it is a species introduced and adapted to the country's climate conditions, it is necessary to conduct preclinical studies demonstrating its anti-inflammatory and antioxidant potential. Objective: Evaluate the effect of a C. longa extract on peritoneal macrophages infected by H. pylori. Methods: With the purpose of describing the anti-inflammatory and antioxidant effect of C. longa on murine macrophages infected by H. pylori, an evaluation was conducted of various extract concentrations and bacterial relationships, while cell death was assessed by DAPI. Determination was made of nitric oxide and hydrogen peroxide production, as well as of interleukin-1β levels. Results: Viability of the macrophage was affected in the presence of 100 µg/ml concentrations of the turmeric extract and as from 25 bacteria / macrophage. When different concentrations of the extract were combined with the bacterial multiplicities, a reduction was observed in H2O2 and IL-1β levels. However, nitric oxide reduction was observed within the range of 6.25-50 µg/ml of the natural product. Conclusions: The Cuban turmeric extract was found to have antioxidant and anti-inflammatory potential by reducing cell cytotoxicity and the production of reactive oxygen species in peritoneal macrophages.

Micro-CT analysis of hyperuricemia on alveolar bone destruction caused by short-term periodontal infection in mice / 口腔疾病防治

Objective@# Establish a murine model for hyperuricemia (HU) and periodontitis to explore whether there is correlation between them and provide a basis for periodontal treatment.@*Methods@#Fourteen male KM mice were divided into 2 groups; the HU group (n=7) was fed food supplemented with potassium oxonate and uric acid, the NC group (n=7) was fed standard food, and the induction period was 35 days. On the 25th day, the molars on one side were ligated to induce periodontitis (P side), while the opposite was true for the control (C side). Baseline and terminal serum uric acid (UA) levels were detected, and alveolar bone resorption was analyzed by micro-CT.@*Results@#The serum UA level of HU mice was (112.94 ± 26.82 )mol/L, that of the NC group was (72.21 ± 19.95) μmol/L, and the difference in UA level was statistically significant (P < 0.05). The P side bone volume fractions of the HU and NC groups were( 29.01 ± 11.09)% and (29.56 ± 15.27)%, respectively, which were not significantly different (t=-0.072, P=0.944). The P side bone mineral densities of the HU and NC groups were(0.53 ± 0.16) g/cm3 and (0.52 ± 0.14) g/cm3, respectively, which were not significantly different (t=0.038, P=0.970). Additionally, there was no correlation between HU or serum UA and alveolar bone resorption (P > 0.05). @* Conclusion @#This research established a murine model for HU and periodontitis, but based on micro-CT analysis of alveolar bone, no relationship between HU or UA levels and periodontitis was found.

Design of ultrahigh-affinity and dual-specificity peptide antagonists of MDM2 and MDMX for P53 activation and tumor suppression

Peptide inhibition of the interactions of the tumor suppressor protein P53 with its negative regulators MDM2 and MDMX activates P53

Research progress on the relationship between the Raf murine sarcoma viral oncogene homolog B gene mutation and lymph node metastasis of papillary thyroid carcinoma / 生物医学工程学杂志

In recent years, with the improvement of the sensitivity of examination equipment and the change of people's living environment and diet, the rate of thyroid cancer has risen rapidly, which has increased nearly five folds in 10 years. The pathogenesis, clinical manifestation, biological behavior, treatment and prognosis of thyroid carcinoma of different pathological types are obviously different. Papillary thyroid carcinoma (PTC) can develop at any age, which accounts for about 90% of thyroid cancer. It progresses slowly and has favourable prognosis, but lymph node metastasis appears easily. Whether PTC is accompanied by lymph node metastasis has an important impact on its prognosis and outcome. The Raf murine sarcoma viral oncogene homolog B(BRAF)gene mutation plays a crucial role in PTC lymph node metastasis. Having an in-depth understanding of the specific role and mechanism of BRAF gene mutation in PTC is expected to provide new ideas for diagnosis and treatment of PTC.

Murine models of primary biliary cholangitis and its role in disease research / 中国药理学通报

Primary biliary cholangitis (PBC) is an autoimmune liver disease characterized by destructive cholangitis affecting the small intrahepatic bile ducts, and the presence of antimitochondrial antibodies (AMA). At present, there has been limited progress in exploring the pathogenesis of the disease, because of difficulty in obtaining human tissue samples and lack of suitable animal models. In recent years, some murine models have been reported, which showed serological, biochemical, and histological characteristics similar to those of humans with PBC. In this review, various murine models of PBC are reviewed, and the characteristics and mechanism of PBC in mice are introduced. Besides, their advantages and limitations in the study of PBC are discussed. It hopes to provide an important theoretical basis for studying the pathogenesis of PBC and developing therapeutic drugs for PBC.

Alterations of DNA damage response pathway: Biomarker and therapeutic strategy for cancer immunotherapy

Genomic instability remains an enabling feature of cancer and promotes malignant transformation. Alterations of DNA damage response (DDR) pathways allow genomic instability, generate neoantigens, upregulate the expression of programmed death ligand 1 (PD-L1) and interact with signaling such as cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) signaling. Here, we review the basic knowledge of DDR pathways, mechanisms of genomic instability induced by DDR alterations, impacts of DDR alterations on immune system, and the potential applications of DDR alterations as biomarkers and therapeutic targets in cancer immunotherapy.

Sensitization with Eustrongylides sp. (Nematoda: Dioctophymatidae) antigens induce production of specific IgG and IgE in murine model / Sensibilização com antígenos de Eustrongylides sp. (Nematoda: Dioctophymatidae) induzindo a produção de IgG e IgE em modelo murino

Abstract Fish consumption plays an important role in human diet. Hoplias malabaricus, commonly known as traíra, is a freshwater fish widely appreciated in several Brazilian states and frequently infected by Eustrongylides sp. fourth-instar larvae (L4). The aim of the present study was to evaluate allergenic potential of Eustrongylides sp. L4 crude extract (CEE). BALB/c mice were immunized intraperitoneally (IP) by 10 μg CEE with 2 mg of aluminum hydroxide on days 0 and 35. Specific IgG and IgE antibody levels were determined after immunization and cellular immunity was evaluated by assessing intradermal reaction in ear pavilion. Epicutaneous sensitization was performed in dorsal region by antigen exposure using a Finn-type chamber containing 50 μg of CEE or saline solution, followed by evaluation of specific antibody levels. IP immunization resulted in a gradual increase in IgG antibody levels and transitory IgE production. Significant increase in ear thickness was observed in cellular hypersensitivity reaction. In case of antigen exposure by epicutaneous route, CEE was able to induce meaningfully increased levels of specific IgG and IgE antibodies as well as heightened cellular immunity. Both intraperitoneal immunization and epicutaneous contact with Eustrongylides sp. larval antigens were observed for first time to be capable of inducing immunological sensitization in mice.

Resumo Consumir peixe constitui papel importante na dieta humana. Hoplias malabaricus, comumente chamado de traíra, peixe de água doce largamente apreciado no Brasil, é frequentemente infectado com larvas de quarto estágio (L4) de Eustrongylides sp. O presente estudo objetivou avaliar o potencial alergênico do Extrato Bruto de L4 de Eustrongylides sp. (EBE). Camundongos BALB/c foram imunizados intraperitonealmente (IP) por 10 μg de EBE com 2 mg de hidróxido de alumínio nos dias 0 e 35. Após imunização, determinaram-se níveis específicos de anticorpos IgG e IgE e avaliou-se a imunidade celular pela reação intradérmica no pavilhão auricular. Realizou-se sensibilização epicutânea na região dorsal pela exposição ao antígeno, utilizando-se câmara tipo Finn, contendo 50 μg de CEE ou solução salina. Após exposições, foram avaliados níveis específicos de anticorpos. Na imunização via IP, houve aumento gradual nos níveis de anticorpos IgG e produção de IgE transitória. Foi observado aumento significativo na espessura do pavilhão auricular na reação de hipersensibilidade celular. Na exposição ao antígeno pela via epicutânea, o EBE induziu aumento significante nos níveis de anticorpos IgG e IgE específicos e induziu imunidade celular. Pela primeira vez, observou-se que a imunização intraperitoneal e contato epicutâneo com antígenos larvares de Eustrongylides sp. são capazes de induzir sensibilização imunológica em camundongos.

Therapeutic effect of human umbilical cord mesenchymal stem cells on imiquimod-induced psoriasis-like mouse models / 中华皮肤科杂志

Objective:To investigate the therapeutic effect of human umbilical cord mesenchymal stem cells (MSCs) on psoriasis-like mouse models induced by imiquimod and the underlying mechanisms.Methods:Eighteen C57BL/6 mice were randomly and equally divided into vaseline group, model group and treatment group according to a random number table. The mice in the model group and treatment group received topical treatment with 5% imiquimod cream at a dose of 62.5 mg once a day for 6 consecutive days on the shaved back, and those in the vaseline group received the treatment with the same amount of vaseline ointment; the mice in the treatment group were injected with 1.5×10 6 human umbilical cord MSCs via the caudal vein on days 1 and 4. The severity of skin lesions on the back of the mice was assessed everyday according to the psoriasis area and severity index (PASI) . Twenty-four hours after the last treatment, that is, on day 7, blood samples were taken, and the mice were sacrificed. The dorsal skin tissues were resected and subjected to hematoxylin and eosin (HE) staining. A single cell suspension of the resected spleen was prepared, and flow cytometry was performed to detect the Th1 and Th17 cell subsets in the spleen cells. Enzyme-linked immunosorbent assay was conducted to detect serum levels of cytokines interleukin (IL) -17A and tumor necrosis factor (TNF) -α. One-way analysis of variance was used for comparisons among groups, Tukey test for multiple comparisons, and repeated measures analysis of variance for the analysis of changes in the PASI score over time. Results:On day 7, there was obvious scaly erythema on the back of the mice in the model group, and the skin thickness and number of infiltrating inflammatory cells were significantly higher in the model group (78.73 ± 23.11 μm, 36.16 ± 2.95 cells/mm 2) than in the vaseline group (13.28 ± 4.57 μm, 13.33 ± 1.15 cells/mm 2, q=19.25, 7.21, respectively, both P < 0.001) . The treatment group showed significantly decreased PASI score, epidermal thickness and number of infiltrating inflammatory cells compared with the model group (all P < 0.001) . The percentage of Th17 cell subsets in the spleen cells and serum level of TNF-α were significantly lower in the treatment group than in the model group (both P < 0.05) . There were no significant differences in the spleen weight, spleen index, spleen cell count, Th1 cell percentage or serum IL-17A level between the treatment group and the model group (all P>0.05) . Conclusion:Human umbilical cord MSCs can effectively alleviate skin inflammation induced by imiquimod in the psoriasis-like mouse models, likely by inhibiting Th17 cell formation and TNF-α expression.

Immunohistochemical expression of p53 and murine double minute 2 protein in odontogenic keratocyst versus variants of ameloblastoma

Introduction: Oncogenes and tumor suppressor genes play a major role in cancer formation, growth, and progression. One of the important findings in this area is that murine double minute 2 (MDM2) oncogene is a negative regulator of wild-type p53. In tumors, expressing wild-type p53, inhibition of MDM2 expression will stabilize p53 and allow it to perform its proapoptotic function, while simultaneously preventing MDM2 from exerting its p53-independent oncogenic effects. The intracellular levels of p53 are tightly regulated by MDM2, as it is a key player in autoregulatory feedback loop under nonstressed conditions. The p53-MDM2 relationship is vital not only for essential functions of the cell, but it also appears to be an integrated part of the complex cellular network which supports the importance of this affair and is a hallmark for its coexistence. Subjects and Methods: This study was designed to identify immunohistochemically the expression of p53 and MDM2 gene using monoclonal antibody in 60 cases of formalin-fixed paraffin-embedded tissue blocks, of which 20 cases were of solid multicystic ameloblastoma (SMA), 20 cases were of odontogenic keratocyst (OKC), and 20 cases were of unicystic ameloblastoma (UA). Results: Immunoexpression of p53 and MDM2 was highest in OKC followed by SMA and was minimum in UA. Further results showed positive correlation between both the molecules. Conclusion: The studied showed that the relationship has a significant role in cancer etiology and progression and therefore is an important topic for future research which should help in the development of new therapeutic agent against cancer

Chemotherapeutic Interaction of Vernonia amygdalina (Delile) Leaf Extract with Artesunate and Amodiaquine in Murine Malaria Model

Aim of the Study: Conventional antimalarial drugs are used concurrently with herbal remediesin malarial endemic developing countries.Vernonia amygdalina is one of such popular herbs used in the treatment of malaria. This study aimed at investigating the antimalarial chemotherapeutic interaction ofVernonia amygdalina (VA) when combined with Amodiaquine (AQ) and/or Artesunate (AS) in a murine Plasmodium berghei malaria model.Methodology:Various doses of aqueous VA leaf extract (100-500 mg/kg/day), AQ (2-10 mg/kg/day) and AS (0.8-4 mg/kg/day) wereadministered orally to P berghei.-infected Swiss albino mice to determine their sub-therapeutic doses. These doses were subsequently used to investigate the chemotherapeutic interactions of VA with AQ and/or AS in both early and establishedmalaria infection test models. The survival of animals with established infections that received different drug/herb treatments were determined using their mean survival time (days) and Kaplan-Meier survival curves (percentage). Using GraphPad Instat (version 3.10) and PrismR(version 5.01) the data obtained were subjected to One-way ANOVA, followed by Student-Newman-Keuls test. P< .05 was considered statistically significant.Results:The sub-therapeutic doses of VA, AQ and AS were found to be 100 mg/kg, 2 mg/kg and 2.4 mg/kg, respectively. The chemosuppressive effect of AQ or AS was significantly increased (p< 0.05) when administered in combination with the VA extract. Similarly, combination of VA extract with AQ or AS resulted in significant (P < .05) parasite clearance when compared to the effects of the herb or the conventional drugs administered separately. The mean survival period of animals with established infection was also significantly enhanced by the VA alone or with AQ(or AS) compared to placebo