A feasible in vitro method to evaluate bacterial infiltration in three implant-abutment connection systems / Método in vitro factible para evaluar la infiltración bacteriana en tres sistemas de conexión implante-pilar

Introduction: Microorganism infiltration through the im-plant-abutment interface causes oral health problems such as periimplantitis, leading to implant loss. Materials and Methods: A feasible new method to quantify the Streptococcus mutans (S. mutans) infiltration through the implant-abutment interface gap is introduced in the present work. Internal hexagon (IH; n = 10), external hexagon (EH; n = 10), Morse taper (MT; n = 10), and a control for each group (n = 1) were tested. Bacteria suspension was prepared at 1.5x108 CFU/mL (CFU: colony forming units), and the implants were individually submerged up to the connection level, allowing the bacteria to contact it. The abutment was removed, and bacteria count was performed. Results: The implant sets were tested under normal bacterial growth and early and late biofilm growth conditions. Colony-forming units per mL were obtained, and the results were compared among groups. Differences in bacterial count between the MT and EH (p<0.001) and the MT and IH (p<0.001) groups were significantly higher in the MT-type implant. There was a significant increment of bacterial infiltration in the MTs submitted to late biofilm growth conditions. EH and IH connections are more effective in preventing bacterial infiltration independent of the growth condition. Conclusions: The proposed methodology is feasible to evaluate the infiltration of microorganisms through the implant-abutment interface.

Introducción: La infiltración de microorganismos a través de la interfaz implante-pilar provoca problemas de salud bucal como la periimplantitis, que conduce a la pérdida del implante. Materiales y Métodos: En el presente trabajo se presenta un nuevo método factible para cuantificar la infiltración de Streptococcus mutans (S. mutans) a través de la brecha de la interfaz implante-pilar. Se probaron el hexágono interno (IH; n = 10), el hexágono externo (EH; n = 10), el cono Morse (MT; n = 10) y un control para cada grupo (n = 1). Se preparó una suspensión de bacterias a 1,5x108 UFC/mL y los implantes se sumergieron individualmente hasta el nivel de conexión, permitiendo que las bacterias entraran en contacto con él. Resultados: Se retiró el pilar y se realizó recuento de bacterias. Los conjuntos de implantes se probaron en condiciones de crecimiento bacteriano normal y de crecimiento temprano y tardío de biopelículas. Se obtuvieron unidades formadoras de colonias por ml y los resultados se compararon entre grupos. Las diferencias en el recuento bacteriano entre los grupos MT y EH (p<0,001) y MT e IH (p<0,001) fueron significativamente mayores en el implante tipo MT. Hubo un incremento significativo de la infiltración bacteriana en los MT sometidos a condiciones tardías de crecimiento de biopelículas. Las conexiones EH e IH son más efectivas para prevenir la infiltración bacteriana independientemente de las condiciones de crecimiento. Conclusión: La metodología propuesta es factible para evaluar la infiltración de microorganismos a través de la interfaz implante-pilar.

Influence of different opacities and layering technique of nanotechnology composite resins regarding the wavelength and fluorescence intensity: in vitro study / Influencia de diferentes opacidades y la técnica de estratificación de resinas compuestas de nanotecnología en la longitud de onda e intensidad de fluorescencia: estudio in vitro

Objective: To evaluate the influence of opacity and the layering technique on the fluorescence of different composite resins. Materials and Methods: Two opacities (enamel and dentin) and the layering technique (enamel + dentin) of the composite resins: Filtek® Z350 and Palfique LX5 were evaluated in vitro. Composite resin discs were fabricated using a preformed matrix of 10 mm diameter and 0.5 mm thick for the single opacity groups and 10 mm thick for the layering technique groups, using 2 layers of 0.5 mm thickness of each opacity (n = 5). Specimens were analyzed using the Raman spectroscopy method. Data were analyzed using the Kruskall-wallis and Mann-Whitney U tests. Results: When evaluating the intensity of fluorescence, no statistically significant difference was found when comparing the layering technique and enamel opacity (p2> 0.05) and an increase in the dentin opacity value for both brands of composite resin. Regarding wavelength, no statistically significant difference was found when comparing the layering technique with enamel opacity and dentin opacity for both Filtek® Z350 and Palfique LX5® composite resins (p2 > 0.05). Conclusions: The fluorescence intensity of the layering technique is similar to enamel opacity for both composite resins. Likewise, the wavelength of the layering technique is similar to the enamel opacity and dentin opacity for both brands.

Objetive: Evaluar la influencia de la opacidad y de la técnica de estratificación en la fluorescencia de diferentes resinas compuestas. Materiales y Métodos: Se evaluó in vitro 2 opacidades (Esmalte y Dentina) y la técnica de estratificación (Esmalte + Dentina) de las resinas compuestas: Filtek® Z350 y Palfique LX5. Se fabricaron discos de resina compuesta, utilizando una matriz preformada de 10 mm de diámetro y 0,5 mm de grosor para los grupos de opacidad única y 10 mm de grosor para los grupos de técnica estratificada, utilizando 2 capas de 0,5 mm de cada opacidad (n = 5). Los especímenes se analizaron mediante el método de Espectroscopía Raman. Los datos se analizaron utilizando la prueba de Kruskall-wallis y Prueba U de Mann Whitney. Resultado: Al evaluar la intensidad de fluorescencia no se encontró diferencia estadísticamente significativa entre los pares: Técnica estratificada versus Opacidad Esmalte para ambas marcas de resina compuesta Filtek® Z350 y para Palfique LX5® (p2 > 0,05). Para longitud de onda no se encontró diferencia estadísticamente significativa entre los pares: Técnica estratificada versus Opacidad Esmalte y Técnica estratificada VS Opacidad Dentina para ambas resinas compuesta Filtek® Z350 y Palfique LX5® (p2> 0,05). Conclusión: La intensidad de fluorescencia de la técnica estratificada es similar a la opacidad Esmalte para ambas resinas compuestas. De igual manera la longitud de onda de la técnica estratificada es similar a la opacidad Esmalte y opacidad Dentina para ambas marcas.

Impacto de la pandemia de COVID-19 en pacientes sometidas a fertilización in vitro en el Hospital Gustavo Fricke / Impact of the COVID-19 pandemic on patients undergoing in vitro fertilization at the Gustavo Fricke Hospital

Introducción: La infertilidad es una enfermedad presente en el 17.5% de la población adulta. La COVID-19 provocó sobrecargas en atención de salud, generando un impacto incierto en unidades de salud reproductiva que realizan terapias de reproducción asistida. Objetivos: Comparar características clínicas y tiempos de atención de pacientes sometidas a fertilización in vitro de la Unidad de Medicina Reproductiva del Hospital Gustavo Fricke entre los periodos 2017-2019 y 2020-2022. Método: Estudio observacional descriptivo, utilizando base de datos anonimizada de la Unidad de Medicina Reproductiva del Hospital Gustavo Fricke con 82 pacientes sometidas a fertilización in vitro. Resultados: La media de edad fue de 34.2 ± 4.2 años. El tiempo de infertilidad fue de 5.1 ± 4.3 años. Los factores de infertilidad más frecuentes son el tuboperitoneal (59,8%) y masculino (46,3%). Durante los primeros 20 meses en prepandemia hubo mayor tendencia a derivación precoz al Centro de Reproducción Humana de la Universidad de Valparaíso (odds ratio: 3.84). Conclusiones: En pandemia se evidenció un aumento en el tiempo de infertilidad, mediana de gestaciones, abortos previos y frecuencia de factor masculino. No hubo cambios significativos en tiempos de derivación a fertilización in vitro, no obstante, durante la pandemia disminuyeron las derivaciones precoces.

Introduction: Infertility is a disease present in the 17.5% of the adult population. COVID-19 caused overload in health centers, producing an unclear impact in the reproductive health units that perform assisted reproduction therapies. Objectives: To compare clinical characteristics and the attention times of patients subjected to in vitro fertilization in the Reproductive Medicine Unit of the Gustavo Fricke Hospital between the 2017-2019 and 2020-2022 periods. Method: Descriptive observational study in which an anonymized database of 82 patients subjected to in vitro fertilization in the Reproductive Medicine Unit of the Gustavo Fricke Hospital was used. Results: The average age was 34.2 ± 4.2 years old. The infertility time was 5.1 ± 4.3 years. The most frequent infertility factors were tuboperitoneal (59.8%) and male factors (46.3%). During the first pre-pandemic 20 months, there was a higher tendency towards early referral to the Center for Human Reproduction of the Valparaiso University (odds ratio: 3.84). Conclusions: In the course of the pandemic, there was an increase of the infertility time, the pregnancies median, prior abortions, and male factor frequency. On the other hand, there were not considerable changes when it comes to referral times to in vitro fertilization, however, early referrals decreased during the pandemic.

Açaí juice stains a glazed resin-modified glass-ionomer cement / Suco de apaí provoca o manchamento do glaze do cimento de ionomero de vidro modificado por resina

ABSTRACT Coloured compounds (anthocyanins) in açaí can stain resin-modified glass-ionomer cement (RMGIC) due to its low staining resistance. Aim The aim of this study was to assess whether açaí compromises the surface colour and roughness of RMGIC in vitro. Materials and Method Disc-shaped specimens (2 mm thick, 8 mm in diameter) of Vitremer™ (3M ESPE, St Paul, MN, USA) were prepared according to the manufacturer 's instructions. The mixture was inserted into a silicone mouldplaced between two mylar strips, and light cured. Specimens were randomly divided into three groups (n=25) according to the solutions to be used for chemical degradation: artificial saliva (control), açaí sorbet and açaí juice. A spectrophotometer CM-2600d/2500d (Konica Minolta, Tokyo, Japan) was used to analyse the colour (CIELa*b* scale). Surface roughness (Ra, mm) was measuredusing theprofilometer Surfcorder SE 1700 (Kosaka Corp, Tokyo, Japan). The specimens were subjected to three daily soaks (6 ml, 15 minutes) for 14 days at 37°C. They were washed in distilled water and placed in fresh saliva (30 minutes in the interval). After the third soak in a day, they were stored in fresh saliva overnight. Outcomes were analysed at baseline (L*, a*, b*, Ra) and after degradation (L'*, a'*, b'*, Ra'). Results The pH values of saliva, sorbet, and juice were 7.0, 3.8, and 4.9, respectively. ΔE* values were 6.6 for saliva, 6.9 for sorbet and 7.8 for juice. There was a significant ΔE* difference between saliva (p=0.005) and juice (p=0.002), and between juice and sorbet (p=0.019), but none between saliva and sorbet (p=0.401). There was no significant Δb* difference between the solutions. No difference between juice and sorbet was observed for Δa*, but they were significantly different from saliva (p<0.001). Brightness (L*) changed significantly. Juice showed the highest ΔE* (7.8) and ΔL* (7.7). No significant change was observed for roughness and there was no difference between the solutions for ARa. Conclusions Açaí and saliva led to unacceptable staining, but no significant roughness changes in the resin-modified glass-ionomer cement.

RESUMO As antocianinas presentes no açaí podem manchar o cimento de ionomero de vidro modificado por resina (CIVMR) devido a baixa resistencia ao manchamento do material. Objetivo O objetivo desse estudo foi avaliar se o açaí compromete a cor e a rugosidade de superficie de um CIVMR in vitro. Materials e Método Amostras (2 mm de espessura, 8 mm de diámetro) de Vitremer™ (3M ESPE, St Paul, MN, USA) foram preparadas de acordo com as instrugoes do fabricante. O materialfoi espatulado, inserido em um molde de silicone colocado entre duas tiras de poliestireno e fotopolimerizado. Após, as amostras foram randomizadas e alocadas em tres grupos (n=25) de acordo com as solugoes usadas para a degradagao química: saliva artificial (controle) e sorbet de açaí e suco de açaí. Utilizou-se o espectrofotometro CM-2600d/2500d (Konica Minolta, Tokyo, Japan) para a análise da cor (escala CIELa*b*) e o rugosímetro Surfcorder SE 1700 (Kosaka Corp, Tokyo, Japan) para a rugosidade de superficie (Ra, mm). As amostras foram submetidas a tres imersoes diárias (6 ml, 15 minutos) em cada solugao por 14 dias a 37°C, tendo sido lavadas em água destilada e mantidas em saliva fresca (30 minutos) nos intervalos. Após a terceira imersao no dia, as amostras foram mantidas em saliva renovada até o dia seguinte. As variáveis foram analisadas antes (L*, a*, b*, Ra) e depois da degradagao química (L'*, a'*, b'*, Ra'). Resultados Os valores de pH da saliva, sorbet e suco foram, respectivamente 7,0, 3,8 e 4,9. Houve diferenga significante para ΔE* entre saliva (p=0.005) e suco (p=0.002) e entre suco e sorbet (p=0.019), mas nao entre saliva e sorbet (p=0.401). Nao foi observada diferenga significante para Δb* entre as solugoes. Nao houve diferenga significante para Δa* entre suco e sorbet, mas eles foram significativamente diferentes da saliva (p<0.001). A luminosidade (L*) mostrou alteragao significante. O suco mostrou os maiores valores de ΔE* (7,8) e ΔL* (7,7)". Nao houve mudanga significante para a rugosidade e nao foi observada diferenga significante entre as solugoes para ARa (p>0.05). Conclusao O açaí e a saliva causaram manchamento inaceitável do glaze do CIVMR e insignificante alteragao da rugosidade.

Comparación de la microdureza de resina compuesta con técnica índex convencional y modificada con cubeta impresa en dos tiempos de fotopolimerizado / Comparison of the microhardness of composite resin with conventional and modified index techniques with printed cuvette in two light-curing times / Comparação da microdureza da resina composta com a técnica de índice convencional e modificada com moldeira impressa em dois tempos de fotopolimerização

RESUMEN Objetivo : Comparar la microdureza de la resina compuesta con técnica índex convencional y modificada con cubeta impresa en dos fotopolimerizaciones. Materiales y métodos : Se contó con seis grupos de estudio de acuerdo a la técnica y a la cantidad de fotopolimerizaciones: técnica directa con una fotopolimerización (D1F), directa con dos fotopolimerizaciones (D2F), índex convencional con una fotopolimerización (IC1F), índex convencional con dos fotopolimerizaciones (IC2F), índex modificada con una fotopolimerización (IM1F), índex modificada con dos fotopolimerizaciones (IM2F). Se utilizaron quince muestras por cada grupo. Las muestras tuvieron dimensiones de 2 mm de altura por 5 mm de diámetro. Se realizó la fotopolimerización siguiendo las indicaciones del grupo al que corresponde, para luego someter a la prueba de dureza de Vickers a tres indentaciones por cada cara con carga de 200 g por 15 segundos. Se realizaron dos registros de microdureza, una superficial y otra a 2 mm. Se recolectaron los datos en el instrumento confeccionado y fueron procesados mediante SPSS v. 26. Resultados : Existe una diferencia significativa entre los grupos de microdureza superficial de primera fotopolimerización (p < 0,001); y también existe diferencia significativa entre los grupos de microdureza a 2 mm de primera fotopolimerización (p < 0,001). Asimismo, no existen diferencias significativas entre los grupos de microdureza superficial de segunda fotopolimerización (p = 0,519) ni en los grupos de microdureza a 2 mm de segunda fotopolimerización (p = 0,279). Conclusiones: No existen diferencias significativas en la microdureza superficial y a 2 mm de profundidad con técnica índex convencional y modificada en cubeta impresa en dos fotopolimerizaciones.

ABSTRACT Objective : To compare the microhardness of composite resin with conventional and modified index techniques with a printed tray in two light cures. Materials and methods : There were six study groups according to the technique and the number of photopolymerizations: direct technique with one photopolymerization (D1F), direct with two photopolymerizations (D2F), conventional index with one photopolymerization (IC1F), conventional index with two photopolymerizations (IC2F), modified index with one photopolymerization (IM1F), modified index with two photopolymerizations (IM2F). Fifteen samples were used for each group. The samples had dimensions of 2 mm in height by 5 mm in diameter. The photopolymerization was performed following the indications of the group to which it corresponds and then subjected to the Vickers hardness test with three indentations on each side with a load of 200 g for 15 seconds. Two microhardness recordings were made, one superficial and the other at 2 mm. Results : There is a significant difference between the first light-curing surface microhardness groups (p < 0.001), and there is also a significant difference between the first light-curing 2 mm microhardness groups (p < 0.001). Likewise, there are no significant differences between the second light-curing surface microhardness groups (p = 0.519) or the second light-curing 2 mm microhardness groups (p = 0.279). Conclusions : There are no significant differences in surface microhardness and microhardness at 2 mm depth with conventional and modified index techniques in printed trays in two photopolymerizations.

RESUMO Objetivo : Comparar a microdureza da resina composta com a técnica de índice convencional e modificada com moldeira impressa em duas fotopolimerizações. Materiais e métodos : Foram constituídos seis grupos de estudo de acordo com a técnica e o número de ciclos de fotopolimerização: técnica direta com um ciclo de fotopolimerização (D1F), direta com dois ciclos de fotopolimerização (D2F), índice convencional com um ciclo de fotopolimerização (IC1F), índice convencional com dois ciclos de fotopolimerização (IC2F), índice modificado com um ciclo de fotopolimerização (IM1F), índice modificado com dois ciclos de fotopolimerização (IM2F). Foram utilizadas quinze amostras para cada grupo. Os espécimes tinham 2 mm de altura por 5 mm de diâmetro. A fotopolimerização foi efetuada de acordo com as indicações do grupo a que corresponde, e depois submetida ao teste de dureza Vickers com três indentações de cada lado com uma carga de 200 g durante 15 segundos. Foram efetuados dois registos de microdureza, um superficial e outro a 2 mm. Os dados foram recolhidos no instrumento e processados utilizando o SPSS v. 26. Resultados : Há uma diferença significativa entre os primeiros grupos de microdureza superficial fotopolimerizável (p < 0,001); e há também uma diferença significativa entre os primeiros grupos de microdureza de 2 mm fotopolimerizável (p < 0,001). Da mesma forma, não há diferença significativa entre os grupos de microdureza da superfície do segundo fotopolimerizador (p = 0,519) e nos grupos de microdureza de 2 mm do segundo fotopolimerizador (p = 0,279). Conclusões : Não existem diferenças significativas na microdureza da superfície e na microdureza a 2 mm de profundidade com a técnica de índice convencional e modificada em duas fotopolimerizações.

Endotoxin test by recombinant Factor C for 0.9% sodium chloride injection

Endotoxin contamination is a threat to the safety of pharmaceutical products, especially parenteral drugs. Any sterile and/or pyrogen-free pharmaceutical product requires regulatory specifications to ensure safe patient use. This study covers the performance evaluation study of an endotoxin quantitation commercial kit by recombinant Factor C (rFC), Endozyme II® Go, for 0.9% sodium chloride injection. The samples were spiked with endotoxin solutions between 0.0005 and 10 EU/mL and tested by the rFC kit to evaluate precision, accuracy, detection and quantification limits, linearity, and robustness. Each of the six points was assayed at least five times.The relative standard deviation for precision testing ranged from 1.9 to 8.3%. The recovery accuracy values of endotoxin were between 61% and 125% for the range from 0.005 to 10 EU/mL. The results demonstrated that the rFC method allows endotoxin quantification with accuracy, precision, specificity, and linearity for the range of 0.005 and 10 EU/mL for 0.9% sodium chloride injection. (AU)

A contaminação por endotoxinas é uma ameaça à segurança dos produtos farmacêuticos, especialmente dos medicamentos parenterais. Qualquer produto farmacêutico estéril e/ou livre de pirogênios requer especificações regulatórias para garantir a segurança de uso para o paciente. Este estudo abrange o estudo de avaliação de desempenho empregando o kit comercial Endozyme II® Go para quantificação de endotoxina, por Fator C recombinante (FCr), em amostras de cloreto de sódio 0,9% para uso parenteral. As amostras foram fortificadas com cinco concentrações distintas de soluções de endotoxina na faixa entre 0,0005 e 10 UE/mL. Cada um dos cinco níveis foi testado pelo menos cinco vezes para avaliação dos critérios de precisão, exatidão, limites de detecção e quantificação, linearidade e robustez. O desvio padrão relativo para os testes de precisão variou de 1,9 a 8,3%. Os valores de recuperação de endotoxina para o parâmetro exatidão estiveram compreendidos entre 61% e 125%. Os resultados demonstraram que o método por FCr permite a quantificação de endotoxinas com exatidão, precisão, especificidade e linearidade para a faixa de 0,005 e 10 UE/mL em amostras de cloreto de sódio 0,9% para uso parenteral. (AU)

Construction and expression of circular RNA in vitro / 中国生物制品学杂志

@#Objective To construct encoding RNA that can be cyclized in vitro by using the permuted intron exon(PIE)strategy in the maturation process of eukaryotic mRNA,and transfect it into HEK-293T cells for expression.Methods The sequences of 5'and 3'cyclic arms with groupⅠcatalytic intron,the internal ribosome entry sites(IRES)of Coxsackievirus B3(CVB3)and the target gene were selected to construct the template plasmid. Linearization plasmid template obtained by PCR was used to synthesize linear RNA through in vitro transcription(IVT),which then started in vitro cyclization(IVC)by the addition of cyclization reagents to obtain circular RNA(circRNA). RNA cyclization was confirmed by agarose gel electrophoresis and ribonuclease R(RNase R)digestion. HEK-293T cells were transfected with circRNAs respectively carrying enhanced green fluorescent protein(EGFP),firefly luciferase(Fluc),and influenza virus hemagglutinin(HA)IVR-180 genes,to verify their expression with in vitro.Results With RNA cyclization,the main band of agarose gel electrophoresis became smaller and small fragments appeared. After RNase R digestion,only some circRNA bands remained.HEK-293T cells transfected with EGFP-circRNA showed significant green fluorescence under the fluorescence microscope.The Fluc expression values of HEK-293T cells transfected with Fluc-circRNA were on average 20 times higher than non cyclized RNA,and the relative light unit(RLU)scaled up with the increase of Fluc-circRNA transfection dose. Western blot analysis showed that HA protein was successfully expressed in HEK-293T cells transfected with HA-circRNA.Conclusion In this study,linear RNA was successfully cyclized in vitro and different proteins were expressed,which lays a foundation of the research of new influenza vaccines and mRNA vaccines.

Establishment and validation of a universal test method in vitro for SARS-CoV-2 mRNA vaccine potency / 中国生物制品学杂志

@#Objective To establish and verify a universal and stable potency test method in vitro for SARS-CoV-2 mRNA vaccine,so as to use it for the quality control of SARS-CoV-2 mRNA vaccine.Methods ELISA kits that could bind well to S protein of SARS-CoV-2 variants,as well as transfected cells,cell plating concentrations and doses for transfection were screened,and then a potency test method for SARS-CoV-2 mRNA vaccine in vitro was established and verified.Results An ELISA kit was found with good binding ability to S protein of each variant,and HEK293T cells were determined as the transfection cells,with the plating concentration of 2.5 × 10~5 cells/mL and the transfection dose of 4 μg/well in the 6-well plate.An universal and stable potency test method for SARS-CoV-2 mRNA vaccine in vitro was established.The verification results showed that the method met the quality control needs.Conclusion The established potency test method in vitro for SARS-CoV-2 mRNA vaccine has good relative accuracy,linearity,intermediate precision and range,and can be applied to the quality control of SARS-CoV-2 mRNA vaccines

Optimization of prescription process of lifitegrast eye drops and evaluation of its efficacy for dry eye disease / 中国药科大学学报

@#In order to develop a more effective drug for dry eye disease, the preparation of lifitegrast eye drops was carried out, and the safety and efficacy of lifitegrast eye drops in vitro and in vivo were investigated. First the method for the determination of lifitegrast content was established, and then the composition and preparation process of the preparation were determined by literature review and single factor experiment. Finally, the safety of lifitegrast eye drops was evaluated by Draize eye irritation test and HE staining, and the therapeutic efficacy was evaluated by Schirmer test and ELISA test. The results showed that the final prescription of lifitegrast eye drops consisted of 5% lifitegrast, 0.4% sodium chloride, 0.3%−0.4% anhydrous disodium hydrogen phosphate, 0.3% sodium thiosulfate pentahydrate and 0.3% sodium hydroxide. The appearance of lifitegrast eye drops was transparent and slightly brownish yellow solution, the pH was7.75±0.05, the osmotic pressure was in the range of 200−330 mOsmol/kg and it had good stability at 60℃ for 3 months. There was no significant difference in irritation study compared with normal saline. Schirmer test showed that tear secretion was increased and the expression of inflammatory factors IL-6, IL-1β and TNF-α in tears were significantly decreased after treatment with lifitegrast eye drops and compared to the commercially available emulsion cyclosporine eye drops, it takes effect faster. The above results indicate that lifitegrast eye drops are simple to prepare and stable, which is a better choice for the rapid treatment of dry eye disease.

Pharmacokinetics and in vitro-in vivo correlation of progesterone suspension injection / 中国药科大学学报

@#Abstract: To investigate the in vitro release, in vivo pharmacokinetics, and the in vitro-in vivo correlation of progesterone suspension injection, self-made progesterone suspension injection was taken as an example. The in vitro release curves of three different particle sizes of progesterone suspension injections were measured using paddle method and dialysis bag method. The in vivo pharmacokinetic characteristics of self-made progesterone suspension injection was studied on SD rats. The plasma concentration of self-made progesterone preparation was detected after intramuscular injection, and correlated with the in vitro release profiles obtained by the dialysis bag method after processing by Wagner-Nelson method. The results showed that when the in vitro release of three different particle sizes of progesterone suspension injections was measured by the paddle method, more than 85% was rapidly released within 20 min, while 85% cumulative release was reached at 40 h, 84 h and 120 h by dialysis bag method, respectively. The release rate obtained by the dialysis bag method was basically consistent with the in vivo release trend, with a correlation coefficient of >0.95, indicating a strong in vivo and in vitro correlation. This study provides some reference for the establishment of the in vitro and in vivo correlation of long-acting suspension injection.

Orrelations of sperm parameters with fertilization rate of in vitro fertilization / 临床检验杂志

Objective To investigate the correction between the fertilization rate of in vitro fertilization(IVF)and sperm parameters.Methods This retrospective study included 327 cases undergoing IVF procedures in the Center for Reproductive Medicine of Chenz-hou First People's Hospital,who were divided into a low fertilization group(fertilization rate＜30％,n=19)and normal fertilization group(fertilization rate≥30％,n=308)based on the assessment standards of quality control data in the embryology laboratory of our center.The basic information of males,embryo information and related sperm parameters were collected,and the correction between sperm parameters and the fertilization rate was analyzed by Pearson correlation.The influencing factors of the fertilization rate were ana-lyzed by binary logistic regression,and the area under the receiver operating characteristic(ROC)curve(AUC)was performed to ana-lyze the predictive value of low fertilization rate.Results The sperm acrosin activity,percentage of spermatozoa with normal morpholo-gy and sperm DNA fragmentation index(DFI)were the relevant factors affecting the fertilization rate,and the correlation coefficients were 0.168,0.306,and-0.243,respectively(all the P＜0.05).Logistic regression analysis showed that all the three indicators were independent factors affecting the fertilization rate of IVF,and the multivariate logistic equation was as follows:estimated fertilization rate=-2.561+0.035 ×acrosin activity+2.066×percentage of sperm with normal morphology-1.51×sperm DFI.The areas under the ROC curve(AUC)of acrosin activity,percentage of sperm with normal morphology,sperm DFI and the combination of them were 0.806,0.889,0.827 and 0.899,respectively.The corresponding cut-off values for the Yuden index were 61.20 μIU/106 sperm,2.98％,27.50％and-21.32,respectively.The sensitivity of acrosin activity,percentage of sperm with normal morphology,sperm DFI and the combination of three indicators for the prediction of low fertilization rate were 95.70％,76.90％,68.40％,64.30％,and their specificity were 43.50％,94.70％,87.70％,100.00％,respectively.The combination of three indicators showed the best prediction ef-ficiency.Conclusion Acrosin activity,percentage of sperm with normal morphology and sperm DFI should be the independent factors to predict the fertilization rate of IVF,and the multivariate logistic equation can be used as a reference to predict the fertilization rate of IVF.

Predictive value of combination of normal morphology sperm percentage and sperm DNA fragmentation index for in vitro fer-tilization embryo transfer outcomes / 临床检验杂志

Objective To investigate the effects of combined assessment of normal morphology sperm rate(MNS)with sperm DNA fragmentation index(DFI)on the outcomes of in vitro fertilization-embryo transfer(IVF-ET).Methods A retrospective analysis was conducted on the clinical data of 641 patients who underwent IVF-ET in our center from June 2020 to February 2023.According to MNS and DFI levels,the patients were divided into group A(MNS＜4％,DFI＜25％,403 cases),group B(MNS ≥4％,DFI ≥25％,9 ca-ses),group C(MNS≥4％,DFI＜25％,125 cases),group D(MNS＜4％,DFI≥25％,104 cases),group A1(MNS＜1％,DFI＜25％,106 cases)and group D1(MNS＜1％,DFI≥25％,60 cases).The general information,semen parameters,embryo develop-ment and pregnancy outcomes were compared among all the 6 groups.Results No statistically significant differences were found be-tween female and male age among these groups(P＞0.05).Significant differences were observed in sperm motility,progressive motili-ty,sperm concentration and fertilization rates(P＜0.05),but no differences of usable embryo rates,good-quality embryo rates,clinical pregnancy rates,early miscarriage rates,or live birth rates(P＞0.05)among the groups A,B,C,D,A1 and D1.Conclusion The decreased MNS and increased DFI in males were associated with reduced sperm motility and concentration,affecting fertilization rates in IVF-ET,but had no significant impact on clinical pregnancy outcomes.The combined assessment of MNS with DFI had limited pre-dictive value for IVF-ET pregnancy outcomes.

Analysis of Cumulative Live Birth Rate of Selective Single Embryo Transfer by Time-lapse Monitoring System and Conventional Morphological Assessment in IVF/ICSI-ET / 实用妇产科杂志

Objective:To analyse the clinical significance of selective single embryo transfer by time-lapse mo-nitoring(TLM)or conventional morphology assessment(CMA)in vitro fertilization/intracytoplasmic sperm in-jection and embryo transfer(IVF/ICSI-ET),and to initially explore the predictive value of Raman spectral analy-sis of embryo culture medium for clinical pregnancy rate.Methods:The study is a prospective randomized con-trolled clinical trial.We assigned 139 patients treated with IVF/ICSI-ET in Reproductive and Genetics Center of Suzhou Municipal Hospital from April 2019 to July 2020,which were randomly assigned to either the CMA or the TLM group.We performed selective single-embryo transfer(fresh cycle and FET)after selecting the optimal em-bryos with TLM or CMA respectively.If the patient's first embryo transfer was unsuccessful,a second one would be performed to compare the differences in the cumulative live birth rate of embryo transfer and other pregnancy outcomes between the two groups.Meanwhile,we collected 15 μl of embryo culture medium at day 3 after IVF/ISCI fertilization for Raman spectroscopy analysis.Results:There were no differences in cumulative live birth,cu-mulative clinical pregnancy,cumulative premature birth,cumulative early spontaneous abortion,cumulative ectopic pregnancy and LGA or SGA between TLM and CMA groups(P＞0.05).The Neonatal sex ratio in the TLM group was lower than that in the CMA group,but the difference was not significant(P＞0.05).Raman spectros-copy analysis of embryo culture medium predicted the clinical pregnancy rate with 67.21％accuracy.Conclu-sions:In young women with a good ovarian reserve,the advantage of using TLM to evaluate embryos is not obvi-ous,so we should remain vigilant that embryo selection based on morphokinetic parameters may affect the sex ratio.Raman spectroscopic analysis of embryo culture medium is not yet able to effectively predict the planting ability of embryos.

Trajectories and influencing factors of post-traumatic stress disorder in women with multiple in vitro fertilization embryo transfer failures / 中国实用护理杂志

Objective:To explore the potential categories of post-traumatic stress disorder (PTSD) trajectories in women with multiple in vitro fertilization-embryo transfer (IVF-ET) failures, and to analyze the effects of different demographic characteristics and psychological factors on the potential categories of PTSD trajectories.Methods:This was a prospective empirical research, from May 2021 to October 2022, women with IVF-ET failure ≥ 2 times in the reproductive department of Shanghai First People′s Hospital from May 2021 to October 2022 were selected as the research objects. Post-traumatic stress disorder civilian version scale was used for 4 follow-ups at 3 d (T1), 10 d (T2), 20 d (T3) after the last transplantation failure and 3 d before the next transplantation cycle (T4). Telephone follow-up and online follow-up were combined to obtain the PTSD level at 4 time points. Potential categories of PTSD score trajectories at four time points were identified using a latent category growth model, and analyze influencing factors using unordered multi classification logistic analysis.Results:Totally 196 IVF-ET women were admitted, aged (29.42 ± 4.13) years. Three PTSD trajectories were fitted in this study, including 82 cases (42%) in non-PTSD group, 61 cases (31%) in mild PTSD group and 53 cases (27%) in elevated PTSD group. Logistic regression analysis showed that age, education level, fertility pressure and marital adjustment level were the predictors of PTSD trajectory in women with multiple IVF-ET failures. Compared with the non-PTSD group, women aged ≥35 years, with lower education level and marital adjustment level were more likely to enter the elevated PTSD group ( OR=4.570, 8.540, 0.949, all P<0.05). Women aged 35 years and with greater reproductive pressure were more likely to enter the mild PTSD group ( OR=3.871, 1.063, both P<0.05). Conclusions:There is group heterogeneity in the trajectories of PTSD in women with multiple IVF-ET failures in the next transplantation cycle. Old age, low education level, high fertility pressure and poor marital adjustment can predict the trajectories of PTSD. Fertility stress and marriage adjustment are changeable variables. Medical staff can relieve women′s fertility pressure through health education and mindfulness intervention, promote a good state of marriage adjustment, and minimize the adverse effects of PTSD on the next cycle of conception.

A study on the relationship between irrational fertility cognition and reproductive life quality of in vitro fertilization-embryo transfer female patients: the mediating effect of psychological resilience / 中国实用护理杂志

Objective:To investigate the relationship between psychologicalresilience and irrational parenthood cognition and fertility life quality in female patients with in vitro fertilization-embryo transfer, and to analyze the mediating effect of psychological resilience on irrational parenthood cognition and fertility life quality, so as to provide guidance for infertility patients to improve their psychological resilience and fertility life quality.Methods:A cross-sectional study method was used to conveniently sample 460 female patients with in vitro fertilization-embryo transfer who were admitted to the Women′s Hospital, School of Medicine Zhejiang University from October 2022 to April 2023.General data questionnaire, Connor-Davidson Resilience Scale, Irrational Parenthood Cognitions Questionnaire and the Fertility Quality of Life were used for investigation.Results:A total of 449 valid questionnaires were retrieved. Patients aged (33.31 ± 4.99) years old. The total scores of irrational parenthood cognition, psychological resilience and the fertility quality of life were (38.73 ± 9.31), (63.19 ± 15.40) and (68.18 ± 11.89) points respectively for female patients with IVF-ET. The total score of irrational parenthood cognition was negatively correlated with the total score of fertility quality of life ( r = - 0.513, P<0.01), and the total score of psychological resilience was positively correlated with the total score of fertility quality of life ( r = 0.424, P<0.01). Pychological resilience played a partial mediating role between irrational parenthood cognition and fertility life quality, with the mediating effect accounting for 28.01% of the total effect. Conclusions:The irrational parenthood cognition of female patients with IVF-ET is at a medium level, and the psychological resilience and the fertility quality of life are at a low level. The irrational parenthood cognition has a direct negative predictive effect on the fertility life quality of patients, while the psychological resilience is a positive predictor of fertility life quality. Irrational parenthood cognition can directly or indirectly affect patients′ fertility life quality through psychological resilience. Clinical medical staff should take effective measures to reduce patients′ irrational parenthood cognition level and improve their psychological resilience, so as to improve patients′ fertility life quality.

Recent advances on the application of deep learning in assisted reproductive technology / 实用医学杂志

Deep learning is a machine learning method in the field of artificial intelligence,which simu-lates the workings of the neural network of the human brain to solve complex problems,and has been used in many important researches and applications in the field of medicine,such as diagnostic imaging,biomedical data pro-cessing,drug research and development,personalized medicine,etc.,which improves the accuracy and efficiency of diagnosis and treatment.In the field of assisted reproduction,deep learning could efficiently identify well-grown embryos,suitable oocytes,or sperms during the intervention process,assisting medical staff to make more accu-rate choices to improve pregnancy rates and reduce the risk of multiple pregnancies.This paper summarizes the latest advances in the application of deep learning in the field of assisted reproduction technology in the past 5 years,and provides an outlook for future research.

Melatonin improves the in vitro developmental competence of benzophenone-3 exposed mouse embryos / 实用医学杂志

Objective To investigate the protective effects of melatonin(MT)on early embryo in vitro development of mice after exposure to benzophenone-3(BP-3).Methods Fertilized mouse oocytes at the synge-neic stage were cultured in KSOM culture medium,0.8 μmol/L BP-3 culture medium,and 1×10-7 mol/L MT + 0.8 μmol/L BP-3 mixed culture medium,respectively.The rescue effect of MT on the early embryos developmental potential of BP-3-exposed mice in vitro was explored by detecting the blastocyst rate,gene transcription level,protein expression level,and the degree of DNA damage in the three groups of embryos.Results MT improved the developmental potential of mouse embryos exposed to BP-3 in vitro.Compared with the control group,MT treatment significantly increased the protein expression of ATP5A and ATP5B and decreased the DNA damage(P<0.05).Furthermore,the transcription levels of antioxidant gene Gpx1 and pluripotency related genes Pou5f1 and Cdx2 were significantly up-regulated in MT-treated blastocysts,and the expression of pro-apoptotic gene Bax was decreased.Compared with the control group,BP-3 treatment enhanced the signal intensity of γ-H2AX in blastocysts(P<0.05),while adding MT could effectively alleviate DSBs(P<0.05).Conclusion The physiological concentration of BP-3 exposure has reproductive toxicity,but the addition of appropriate con-centration of MT could significantly improve the in vitro developmental potential and quality of BP-3-exposed early embryos.

In vitro degradation behavior of Mg-Zn-Ca alloys / 中国组织工程研究

BACKGROUND:Due to the complex physiological environment of the human body,a wide variety of simulated physiological fluids have been chosen for the current degradation experiments.Therefore,it is of great interest to analyze the degradation behavior of Mg-Zn-Ca alloys in different simulated body fluid environments. OBJECTIVE:To investigate the degradation process and property changes of Mg-Zn-Ca alloy in different simulated body fluids,and to clarify the influence of Ca content and simulated body fluid type on the alloy. METHODS:Mg-Zn-Ca alloys with calcium content of 0.2%,0.5%and 1%were prepared by melting extrusion molding process and were named Mg-Zn-0.2Ca,Mg-Zn-0.5Ca and Mg-Zn-1Ca alloys in turn,with Mg-Zn alloy as the control.The prepared alloys were placed into three simulated body liquids(physiological saline,PBS and Hank's solution),and the morphology,compositional changes,mass loss,pH value and mechanical properties were characterized and analyzed during the degradation. RESULTS AND CONCLUSION:(1)With the extension of degradation time,a large number of nanoscale lamellae and columnar structures were generated on the surface of the degraded alloy,and the main components were MgO and Mg(OH)2.The degradation rate of the four kinds of alloys in physiological saline was the fastest,and that in Hank's solution was the slowest.The degradation rate in physiological saline was as follows:Mg-Zn<Mg-Zn-0.2Ca<Mg-Zn-0.5Ca<Mg-Zn-1Ca.The degradation rate in PBS and Hank's solution was as follows:Mg-Zn<Mg-Zn-0.2Ca ≈ Mg-Zn-0.5Ca<Mg-Zn-1Ca.(2)With the extension of degradation time,all four kinds of alloys had a certain mass loss.The degradation in physiological saline was the fastest;the degradation in Hank's solution and PBS was slow,and in the same simulated body fluid,with the increase of calcium content in the alloy,the corrosion rate of the alloy was obviously accelerated.(3)The pH rise was mainly concentrated in 1 day and slowed down after that,and the pH change was the largest in PBS.In the same simulated body fluid,with the increase of calcium content in the alloy,the pH value in the degradation environment increased significantly.(4)In the initial state,the elastic modulus of all Mg-Zn-Ca alloys was higher than that of Mg-Zn alloys.After being placed in simulated body fluids,the elastic modulus of the four alloys decreased with the extension of degradation time,and the decrease was most obvious in physiological saline.(5)In conclusion,a small amount of Ca addition improved the mechanical properties of Mg-Zn-Ca alloy.A small amount of Ca does not accelerate the degradation rate of the alloy,but excessive Ca accelerates the degradation rate of the alloy.During the degradation,the effect of physiological saline simulated body fluid on the mechanical strength of the alloy was the most significant.

Bibliometric analysis of researches on liver organoids / 中国组织工程研究

BACKGROUND:In recent years,the development of liver organoids has made it a hot spot in the field of international liver disease research,but there is still no article on the bibliometric analysis of liver organoids. OBJECTIVE:To explore the hot trends in liver organoids in the last 20 years based on bibliometrics and visualization analysis. METHODS:We searched the articles about liver organoids in the Web of Science Core Collection from January 1,2002 to November 12,2022.Origin,Office,and CiteSpace software were used for bibliometrics and visualization analysis.We statistically analyzed the number of annually published articles,countries,institutions,authors,journals,and keywords of the articles by generating charts. RESULTS AND CONCLUSION:The number of articles,citation frequency,institutions and personnel involved in the research about liver organoids showed an overall upward trend in the last 20 years,indicating that the field was growing rapidly and attention was increasing.The USA had published the most papers and had the strongest influence in this field.Although it had invested a lot of time and energy,the number of papers published by a single research institution in the USA was not the highest among many research institutions.China was second only to the USA in the number of publications,with the Chinese Academy of Sciences and Fudan University leading the list.Utrecht University in the Netherlands was the institution with the most publications.Clevers H was the author with the highest number of articles.The article with the highest co-citation frequency was"Long-term culture of genome-stable bipotent stem cells from adult human liver".The main fields of study for liver organoids were Molecular Science,Biology,and Immunology.The most frequently occurring keywords were stem cell,in vitro,and culture.The research hotspots in the liver organoids field were mainly focused on in vitro stem cell three-dimensional culture,differentiation and gene expression.

Abnormal types of intervertebral disc structure and related mechanical loading with biomechanical factors / 中国组织工程研究

BACKGROUND:The problem of intervertebral disc injury and degeneration has been studied in many ways.Many studies have shown that intervertebral disc injury and degeneration is driven by mechanical loading factors.However,the potential relationship between common phenotypes of intervertebral disc injury and degeneration and mechanical loading factors has been rarely summarized. OBJECTIVE:To summarize the types of common structural abnormalities exhibited by intervertebral disc injury and degeneration in the published literature,and sum up the potential links to the types of mechanical loading that lead to these structural abnormalities in in vitro and ex vivo experimental studies. METHODS:Using the terms"intervertebral disc failure,intervertebral disc injury,mechanical load,mechanical factor,load factor,biomechanics"as Chinese and English key words in PubMed,CNKI,and WanFang databases,articles related to intervertebral disc injury degeneration and mechanical load factors were retrieved.Literature screening was performed according to the inclusion and exclusion criteria,and 88 articles were finally included. RESULTS AND CONCLUSION:(1)Common structural abnormalities of intervertebral discs include decreased intervertebral disc height,disc bulge,osteophyte formation,annulus fibrosus tear,intervertebral disc herniation or disc prolapse,endplate damage,Schmorl nodes and intervertebral disc calcification.Intervertebral discs are susceptible to mechanical load types such as compression,bending,axial rotation,and compound loads.(2)The compressive load mainly causes the decrease of the proteoglycan content and the water-binding ability of the intervertebral disc,leading to the decrease or swelling of the intervertebral disc and further damage and degeneration of the intervertebral disc.In addition,the excessive compressive load causes greater damage to the endplate.(3)Bending load and axial rotation load damage the annulus fibrosus more than the endplate,and prolonged or repeated bending loads can cause tearing of the fibrous annulus and herniation or prolapse of the intervertebral disc,while pure axial rotation loads can induce less damage to the intervertebral disc and only cause the tear of the annulus fibrosus.(4)However,when different load types act in combination,it is more likely to result in high stress on the disc and a greater risk of disc injury.(5)Injury and degeneration of the intervertebral disc present progressive structural damage,and early prevention and protection are particularly important in clinical practice.Future tissue engineering research can start with early repair of the intervertebral disc.