ABSTRACT The objective of this study was to identify differentially expressed genes (DEG) between females and males in early bovine embryos. Superovulated cattle (n = 4) in the age span of 16–18 months were artificially inseminated with semen from the same bull that has been proven to be fertile. Blastocysts were collected by routine non-surgical uterine flushing on day 7 after insemination. This study determines the sex of embryos using a micro-injection pipette to collect a few blastomeres through the Zona pellucida. The remaining blastomeres were used for single-cell RNA sequencing (scRNA-seq) analysis on the Illumina platform, followed by differential expression analysis, Gene Ontology (GO) function analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. The criteria for identifying DEG will be set at |log2(Fold change)| ≥ 1 and P<0.05. A total of 8004 DEG were detected in female (Denoted as BLX, n = 3) and male (Denoted as BLY, n = 3) blastocysts. Transcripts highly expressed in the female embryos were related to catalytic activity, nucleotide binding, and catabolic process, while transcripts highly expressed in the male embryos were linked to oxidative phosphorylation, mitochondrion, and the ribosome. Nine genes that may be involved in blastocyst growth and development were screened, suggesting that their differential expression may be responsible for the differences in the development of male and female embryos. This study provides important information on potential genes and pathways associated with differences in early female and male embryonic development.

Abstract Tongluo-Qutong rubber plaster (TQRP), a typical Chinese patent medicine that contains 13 different herbal remedies, is widely used in clinical practice for the treatment of cervical spondylosis and osteoarthritis. However, due to a lack of in vitro transdermal studies, the active ingredients of TQRP have not been fully elucidated. This presents a huge obstacle for quality evaluation, pharmacokinetic studies and clinical safety assessment of TQRP. In this work, a UPLC/UV/MS/MS method was established and validated to evaluate five analytes in TQRP. The validation demonstrated linearity (r > 0.99), specificity (no co-eluting peaks at the retention times of the analytes), and precision (RSD < 15%) within acceptable parameters. A skin permeation study was performed to determine the concentrations of drugs delivered to the dermis. The 24-hour cumulative permeation of ferulic acid, aleo-emodin, emodin and piperine were 303.68, 709.31, 671.06 and 25561.01 ng/cm2, respectively. According to the fitting data of the TQRP active components, skin permeation was mainly due to a combination of passive diffusion and drug release after matrix erosion.

Abstract To analyze value of PDCA circulation mode combined with continuing nursing guided by WeChat on improving the nursing quality of patients with gynecological inflammatory diseases during out-of-hospital treatment. 320 patients with gynecological inflammation were equally divided into control group (routine guidance during out-of-hospital treatment) and observation group (the control group implemented PDCA circulation mode combined with WeChat guidance of continuing care services). Nursing quality score and total nursing efficiency were compared. The patients’ average age, marital status, menopause and disease types showed no difference between two groups. After 3 months of nursing intervention, the scores of nursing quality and total effective rate of nursing in the observation group were better than those in the control group. The improvement degree of mastery rate of gynecological health knowledge and the total nursing satisfaction in the observation group were better than those in the control group. During out-of-hospital treatment of patients with gynecological inflammatory diseases, the implementation of PDCA circulation mode combined with the continuous nursing intervention guided by WeChat is conducive on improving the quality of nursing, improving the grasp of gynecological health knowledge, thus increasing the satisfaction of patients with nursing services, with a high value of promotion and application.

RESUMO OBJETIVO ADAMTS4 é um membro da família ADAMTS4, que secreta proteinases. O mecanismo da metástase do tumor pode ser correlacionado a sua promoção da angiogênese. Determinou-se se ADAMTS4 participa na progressão do câncer colorretal. Métodos A expressão em amostras clínicas e linhas de células CRC foi investigada. Usando a imuno-histoquímica (IHC), a hibridação fluorescente in situ (HFIS) e o RT-PCR, a expressão de ADAMTS4 foi determinada em tumores colorretais de diferentes estágios do câncer e locais anatômicos, e em três linhas de células de níveis de agressividade distintos. Resultados A superexpressão de ADAMTS4 foi observada em amostras de tecido por IHC, e esta foi localizada principalmente no citoplasma, como detectado pelo HFIS. O qRT-PCR e a análise de wester blot corroboraram os resultados clínicos da amostra. Conclusão Os dados atuais corroboram a noção de que a superexpressão de ADAMTS4 no CRC pode ser útil como um biomarcador não invasivo para a detecção de CRC em pacientes.

SUMMARY OBJECTIVE ADAMTS4 is a member of the ADAMTS4 family, which secretes proteinases. The mechanism of tumor metastasis may be correlated to its promotion of angiogenesis. It was determined whether ADAMTS4 participates in colorectal cancer progression. Methods The expression in clinical samples and CRC cell lines was investigated. Using immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), and RT-PCR, the expression of ADAMTS4 was determined in colorectal tumors of different cancer stages and anatomic sites, and in three cell lines of different aggressiveness. Results The overexpression of ADAMTS4 was observed in tissue samples by IHC, and this was mainly located in the cytoplasm, as detected by FISH. The qRT-PCR and western blot analyses further supported the clinical sample findings. Conclusion The present data support the notion that the overexpression of ADAMTS4 in CRC might be useful as a non-invasive biomarker for detecting CRC in patients.

OBJETIVO: Descrever o Single Photon Emission Microscope (SPEM), o estado da arte em tecnologia para SPECT de pequenos animais, e ilustrar seu desempenho com imagens típicas de diferentes órgãos de animais. MÉTODOS: O SPEM consiste em dois dispositivos de imagem independentes que usam cintiladores de alta resolução, câmaras CCD de altas sensibilidade e resolução, e colimadores multiorifício. Durante a aquisição das imagens, o camundongo é posicionado dentro de um tubo vertical rotatório entre os dispositivos imageadores. Posteriormente, um software apropriado, baseado no algoritmo de Máxima Verossimilhança, produz iterativamente a imagem tomográfica. Utilizamos radiofármacos apropriados para imageamento de rins, tireoide, coração e cérebro. Os camundongos foram injetados com 74-148 MBq/0,3mL e foram registrados por 40 a 80 minutos, entre 30 a 60 minutos pós-injeção. Durante esse processo, os animais permaneceram anestesiados com quetamina/xilazina. RESULTADOS: Apresentamos imagens tomográficas de diferentes órgãos de camundongos, que indicam a capacidade do instrumento na aplicação proposta. CONCLUSÃO: O SPEM é uma tecnologia inovadora de SPECT para pequenos animais, que fornece imagens de alta resolução com sensibilidade apropriada para estudos pré-clínicos. Seu uso com radiofármacos apropriados permitirá a realização de estudos translacionais de vários modelos animais de doenças humanas e seu tratamento farmacológico, bem como o desenvolvimento de novos agentes terapêuticos.

OBJECTIVE: To describe the Single Photon Emission Microscope (SPEM), a state-of-the-art instrument for small animal SPECT imaging, and characterize its performance presenting typical images of different animal organs. METHODS: SPEM consists of two independent imaging devices based on high resolution scintillators, high sensitivity and resolution Electron-Multiplying CCDs and multi-pinhole collimators. During image acquisition, the mouse is placed in a rotational vertical holder between the imaging devices. Subsequently, an appropriate software tool based on the Maximum Likelihood algorithm iteratively produces the volumetric image. Radiopharmaceuticals for imaging kidneys, heart, thyroid and brain were used. The mice were injected with 74 to 148 MBq/0,3mL and scanned for 40 to 80 minutes, 30 to 60 minutes afterwards. During this procedure, the animals remained under ketamine/xilazine anesthesia. RESULTS: SPEM images of different mouse organs are presented, attesting the imaging capabilities of the instrument. CONCLUSION: SPEM is an innovative technology for small animal SPECT imaging providing high resolution images with appropriate sensitivity for pre-clinical research. Its use with appropriate radiotracers will allow translational investigation of several animal models of human diseases, their pharmacological treatment and the development of potential new therapeutic agents.