Transcriptomic Analysis of Wuzi Yanzongwan on Testicular Spermatogenic Function in Semi-castrated Male Mice / 中国实验方剂学杂志

ObjectiveTo screen out the transcriptomes related to the intervention of Wuzi Yanzongwan on the spermatogenic function of semi-castrated male mice， and to explore its potential mechanism in the intervention of the progress of low spermatogenic function. MethodBalb/c mice were randomly divided into sham-operated group， model group， testosterone propionate group（0.2 mg·kg-1·d-1， intramuscular injection） and Wuzi Yanzongwan group（1.56 g·kg-1·d-1， intragastric administration） according to body weight， with 12 mice in each group. The right testicle and epididymis were extracted from the model group and the drug administration group to construct the semi-castrated model of low spermatogenic function， while the fur and the right scrotum of the sham-operated group were only cut and immediately sterilized and sutured. At the end of the intervention， hematoxylin-eosin（HE） staining was used to observe the histopathology of testis， enzyme-linked immunosorbent assay（ELISA） was used to detect the levels of serum testosterone（T）， luteinizing hormone（LH） and follicle stimulating hormone（FSH）. The sperm count and motility of epididymis were measured by automatic sperm detector of small animal. Transcriptomic microarray technology was used to detect the mRNA expression level of testicular tissue in each group， the transcriptome of genes related to the regulation of Wuzi Yanzongwan was screened， and three mRNAs were selected for Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） to verify the transcriptome data. Through the annotation analysis of Gene Ontology（GO） and the signaling pathway analysis of Kyoto Encyclopedia of Genes and Genomes（KEGG）， the related functions of drugs regulating transcriptome were analyzed. ResultCompared with the sham-operated group， the testicular tissue of mice in the model group showed spermatogenic injury， contraction and vacuolization of the seminiferous tubules， reduction of spermatogenic cells at all levels， widening of the interstitial space， obstruction of spermatogonial cell development and other morphological abnormalities， and serum T significantly decreased， LH significantly increased（P<0.01）， and FSH elevated but no statistically significant difference， the count and vitality of epididymal sperm significantly decreased（P<0.01）. There were 882 differentially expressed mRNAs in the testicular tissues， of which 565 were up-regulated and 317 were down-regulated. Cluster analysis showed that these differentially expressed mRNA could effectively distinguish between the sham-operated group and the model group. Compared with the model group， the damage to testicular tissue in the Wuzi Yanzongwan group was reduced， the structure of the seminiferous tubules was intact， vacuolization was reduced， and the number of spermatogenic cells at all levels was significantly increased and arranged tightly. The serum T significantly increased， LH significantly decreased（P<0.01）， and FSH decreased but the difference was not statistically significant. The count and vitality of sperm in the epididymis were significantly increased（P<0.01）. Moreover， Wuzi Yanzongwan could regulate 159 mRNA levels in the testes of semi-castrated mice， of which 32 were up-regulated and 127 were down-regulated， and the data of the transcriptome assay was verified to be reliable by Real-time PCR. GO and KEGG analysis showed that the transcriptome functions regulated by Wuzi Yanzongwan were involved in the whole cell cycle process of sperm development such as sex hormone production of interstitial cells in testis， renewal， differentiation， metabolism， apoptosis and signal transduction of spermatogenic cells， and were closely related to the biological behaviors of signaling pathways such as spermatogenic stem cell function， endoplasmic reticulum protein processing and metabolic program. ConclusionWuzi Yanzongwan can effectively improve the low spermatogenic function of semi-castrated male mice， and its mechanism may be related to the regulation of testicular transcriptional regulatory network， the synthesis of sex hormones in testicular interstitial cells， the function of spermatogenic stem cells， the whole cell cycle process of spermatogenesis， as well as the expression of endoplasmic reticulum protein processing and metabolic program related genes transcription.

Design, synthesis, and antifungal mechanism of carbaline fluorescent probes / 药学学报

Three carboline fluorescent probes F1-F3 were designed and synthesized, based on lead compound JYJ-19, an antifungal compound discovered previously by our group. The antifungal activity in vitro results showed that compound F1 had moderate antifungal activity (MIC80 = 32 μg·mL-1). The stokes shift of F1 is 70 nm. The fluorescent probe F1 has good optical properties and can be used for fluorescence imaging research. Subcellular localization experiments results showed that F1 was enriched in the mitochondria of fungal cells. The detection of intracellular reactive oxygen species levels shows that JYJ-19 enhances intracellular reactive oxygen species levels. The above results indicated that carboline compounds could exert antifungal effects by acting on fungal mitochondria.

Biosensor analysis technology and its research progress in drug development of Alzheimer's disease / 药学学报

Biosensor analysis technology is a kind of technology with high specificity that can convert biological reactions into optical and electrical signals. In the development of drugs for Alzheimer's disease (AD), according to different disease hypotheses and targets, this technology plays an important role in confirming targets and screening active compounds. This paper briefly describes the pathogenesis of AD and the current situation of therapeutic drugs, introduces three biosensor analysis techniques commonly used in the discovery of AD drugs, such as surface plasmon resonance (SPR), biolayer interferometry (BLI) and fluorescence analysis technology, explains its basic principle and application progress, and summarizes their advantages and limitations respectively.

Nationwide expert consensus on the clinical application and evaluation of 4K fluorescence high-end medical endoscopes in thoracic surgery (2023) / 中国胸心血管外科临床杂志

@#his consensus aims to introduce the applications of 4K high-definition technology and fluorescence technology in thoracic surgery, summarize and categorize the technical support for pulmonary segment surgery, and innovatively propose technical support for precise sleeve resection of pulmonary segments. It provides a reference for clinical use, points out the direction for the research and innovation of domestically produced high-end endoscopes, promotes the widespread application of excellent domestically produced medical endoscopes, and facilitates the development of domestically produced medical equipment.

Preparation of the first national reference standard for determination of HEK293 cell DNA content / 中国生物制品学杂志

@#Objective To prepare a national reference standard for the quantification of HEK293 cell DNA content,so as to provide a support for the determination of residual DNA in HEK293 cells in the industry.Methods HEK293 cell DNA prepared using Genomic-tip 500/G and genomic DNA purification reagents was used as source materials,and the purity and content were assessed using ultraviolet spectrophotometry and agarose gel electrophoresis.After dilution to approximately 100 ng/μL,the DNA was aliquoted at 160 μL/tube.Five different laboratories were organized for collaborative calibration by using ultraviolet spectrophotometry, and the stability and applicability were evaluated.Results The HEK293 cell DNA national reference standard exhibited A_(260)/A_(280) ratios between 1.8 and 2.0 and displayed a single band on electrophoresis,meeting the specified criteria.Collaborative calibration across five laboratories yielded 78 valid data points with an average content of 104.8 ng/μL,a relative standard deviation(RSD) of 4.2%.The 95% confidence interval for the mean was 103.8—105.8 ng/μL,and the 95% reference range for single measurements was 96.0—113.6 ng/μL.The average confidence limit rate was 1.0%,and the recommended storage condition was-80 ℃.Applicability studies were conducted using two different models of fluorescence quantitative PCR instruments.The reference standard exhibited good applicability within the range of 0.3—3 000 pg/reaction,with amplification efficiencies of 101% and 95%,and R~2 values of 0.999 2 and 0.999 5 for the standard curves,respectively.Conclusion This batch of HEK293 cell DNA national reference standard meets all required specifications and can be utilized as a national reference standard for fluorescence quantitative PCR detection,with a certified content of 104.8 ng/μL,assigned batch number 270039-202301.

Direct spectrofluorimetric methods as alternatives to compendial ones used for the quality control of biopharmaceuticals: development, validation and application

Abstract A simple, rapid, precise, accurate and sustainable spectrofluorimetric method (SFM) was developed, validated and applied for the determination of 4-aminobenzoic acid and aromatic amino acids (phenylalanine, tryptophan and tyrosine). These compounds are used in biopharmaceutical formulations and therefore must be analyzed by quality control laboratories to meet the criteria established in pharmacopoeias. In general, potentiometric titration (PT) is described in the compendia as the official analytical technique. However, this method showed low sensitivity and selectivity, and moreover was performed with a non-aqueous solvent (acetic acid), which led to higher consumption of reagents and consequently to the formation of residues. Therefore, the SFM was developed in aqueous medium at pH 7.2 using phosphate buffer. It was successfully validated according to the ICH guidelines and showed good linearity range (r>0.999), specificity, accuracy and precision (within and between days) and robustness. The test results were compared between the SFM and PT using raw material samples, while according to the F- and t-tests at 95% confidence level, no statistical difference was found between the methods.

Dried Salivary Spot As A Diagnostic Aid- A Review

Saliva is a colourless, watery and complex fluid secreted by the salivary glands into the oral cavity. It plays an important role in maintaining the moisture and well being of the oral cavity. In recent years, Saliva has been used by the researches, as it is non invasive technique, limited training required, potentially valuable for children and elderly patients, cost effective, eliminates the risk of infection and screening in large population. Collecting dried salivary sample is still being a greater challenge in the forensic. The process of identifying a disease, condition, or injury from its signs and symptoms is referred as diagnosis and diagnostic tool helps in simplifying the clinical findings to obtain a reasonable and relevant differential diagnosis. Recently, Dried salivary spot has been most commonly used among Forensic Odontology which can be used as an efficient diagnostic aid. This article aims to review the updates and significance of dried saliva spot and sampling in forensic odontology.

Eventos de blanqueamiento del coral Porites lobata (2006-2022) y actividades de restauración en el sitio Ramsar ''Complejo Los Cóbanos", El Salvador

Introducción: El blanqueamiento global coralino refleja la fragilidad del coral a condiciones ambientales adversas y al cambio climático. Porites lobata es la especie de coral hermatípico más abundante actualmente en El Salvador y ha experimentado eventos de blanqueamiento recientes en el área natural protegida Complejo Los Cóbanos. Objetivo: Describir los episodios de blanqueamiento que colonias de P. lobata experimentaron en Los Cóbanos, El Salvador, durante los últimos 18 años, y analizar los esfuerzos de restauración de coral. Métodos: Aproximadamente 2 100 imágenes (2006 - 2022) tomadas sin periodicidad específica de dos sitios fueron revisadas. Parámetros del agua desde 2014 fueron registradas sin una periodicidad temporal definida. Las áreas de estudio fueron son playa El Faro y colonias submareales cerca de un importante resort de playa. La primera área presentó colonias dispersas en una zona intermareal, la otra, colonias más grandes a 2 m de profundidad en marea baja. También, revisar literatura acerca de actividades para restaurar arrecifes dentro del país. Resultados: Varios eventos de blanqueamiento ocurrieron durante este período de observación. Algunos de ellos podrían estar asociados a temperaturas altas del agua de mar (> 30 o C) de mayo a octubre (estación lluviosa), floraciones algales nocivas, así como causas desconocidas. Varios blanqueamientos que causaron pérdida de cobertura de coral en las colonias intermareales ocurrieron durante el evento global (2015-2016). Conclusiones: Blanqueamiento en el Complejo los Cóbanos ocurre en diferentes patrones de intensidad durante la estación lluviosa que comprende niveles de alta temperatura y turbidez en el agua, así como floraciones algales. El episodio de 2015-2016 causó pérdida de cobertura de coral en las colonias intermareales y las submareales experimentaron pequeños daños permanentes (< 5 % mortalidad). Las políticas de restauración nacionales requieren más esfuerzos estratégicos considerando la hidrodinámica y las capacidades institucionales locales.

Introduction: Global coral bleaching reflects coral fragility to adverse environmental conditions and climate change. Porites lobata is the most abundant hermatypic coral species currently in El Salvador and has experienced recent bleaching events at natural protected area Complejo Los Cóbanos. Objective: To describe the bleaching episodes that colonies of P. lobata experienced at Los Cóbanos, El Salvador, during the last 18 years, and analyze coral restoration efforts. Methods: Approximately 2 100 images (2006 - 2022) taken without specific periodicity from two sites were reviewed. Water parameters since 2014 were registered without defined frequency. The areas sampled were El Faro beach and subtidal colonies near an important beach resort. The first area presented dispersed colonies on an intertidal zone, the other, larger colonies at 2 m deep in low tide. Also, literature about activities to restore reefs within the country was reviewed. Results: Several bleaching events occurred during this observation period. Some of them could be associated to high sea water temperatures (> 30 o C) from May to October (rainy season), others to harmful algae blooms, as well as unknown causes. Severe bleaching that caused loss of coral coverage on intertidal colonies occurred during the global event (2015-2016). Conclusion: Bleaching at Los Cóbanos occurs in different intensity patterns usually during the rainy season that encompasses high temperature and turbidity levels in water, as well as algal blooms. The 2015-2016 episode caused coral coverage loss on the intertidal colonies and subtidal ones experienced little permanent damages (< 5% mortality). National restoration policies for coral colonies require more strategic efforts considering the local littoral hydrodynamics and institutional capabilities.

Comparative Analysis of the Elementary and Amino Acid Composition of Nepeta olgae Regel L. Plants Growing in Uzbekistan

This article for the first time presents the results of the study of qualitative and quantitative elemental and amino acid composition of the aboveground part of the plant Nepeta olgae Regel (L.) taken in the territory of Chust and Kosonsai districts (from the slopes of Gova and Kosonsai mountains) of Namangan region during the period before and during flowering (May-June, 2021-2022). The use of instrumental analysis of high-throughput energy dispersive X-ray fluorescence spectrometry, allowed to establish 20 mineral elements in the plant Nepeta olgae Regel (L.), among which to vital 9 elements and 3 to conditionally necessary. The amino acid composition of the plant Nepeta olgae Regel (L.) was studied by high performance liquid chromatography (HPLC) and 17 compounds were identified. Of these, 8 were substitutable and 9 essential amino acids.

Study of resveratrol against bone loss by using in-silico and in-vitro methods / Estudo do resveratrol contra a perda óssea usando métodos in silico e in vitro

By applying the in-silico method, resveratrol was docked on those proteins which are responsible for bone loss. The Molecular docking data between the resveratrol and Receptor activator of nuclear factor-kappa-Β ligand [RANKL] receptors proved that resveratrol binds tightly to the receptors, showed the highest binding affinities of −6.9, −7.6, −7.1, −6.9, −6.7, and −7.1 kcal/mol. According to in-vitro data, Resveratrol reduced the osteoclasts after treating Marrow-Derived Macrophages [BMM] with Macrophage colony-stimulating factor [MCSF] 20ng / ml and RANKL 50ng / ml, with different concentrations of resveratrol (2.5, 10 μg / ml) For 7 days, the cells were treated with MCSF (20 ng / ml) and RANKL (40 ng / ml) together with concentrated trimethyl ether and resveratrol (2.5, 10 μg / ml) within 12 hours. Which, not affect cell survival. After fixing osteoclast cells with formaldehyde fixative on glass coverslip followed by incubation with 0.1% Triton X-100 in PBS for 5 min and after that stain with rhodamine phalloidin staining for actin and Hoechst for nuclei. Fluorescence microscopy was performed to see the distribution of filaments actin [F.actin]. Finally, resveratrol reduced the actin ring formation. Resveratrol is the best bioactive compound for drug preparation against bone loss.

Com a aplicação do método in-silico, o resveratrol foi ancorado nas proteínas responsáveis pela perda óssea. Os dados de docking molecular entre o resveratrol e o ligante do receptor ativador do fator nuclear kappa-Β [Receptor Activator of Nuclear Factor kappa-B Ligant (RANKL)] provaram que o resveratrol se liga fortemente aos receptores, mostraram as afinidades de ligação mais altas de −6,9, −7,6, −7,1, −6,9, - 6,7 e -7,1 kcal / mol. De acordo com dados in-vitro, o resveratrol reduziu os osteoclastos após o tratamento de macrófagos derivados da medula óssea [Bone Marrow derived Macrophage (BMM)] com fator estimulador de colônias de macrófagos [Macrophage Colony-Stimulating Factor (MCSF)] 20ng / ml e RANKL 50ng / ml, com diferentes concentrações de resveratrol (2,5, 10 μg / ml). Durante sete dias, as células foram tratadas com MCSF (20 ng / ml) e RANKL (40 ng / ml) juntamente com éter trimetílico concentrado e resveratrol (2,5, 10 μg / ml) em 12 horas, processo que não afeta a sobrevivência celular. Após a fixação de células de osteoclastos com fixador de formaldeído em lamela de vidro seguido de incubação com 0,1% Triton X-100 em PBS por 5 min, foi realizado posteriormente o procedimento para corar com rodamina faloidina a actina e Hoechst os núcleos. A microscopia de fluorescência foi realizada para ver a distribuição dos filamentos de actina [F.actina]. Finalmente, o resveratrol reduziu a formação do anel de actina. O resveratrol é o melhor composto bioativo para o preparo de medicamentos contra a perda óssea.

Shading minimizes the effects of water deficit in Campomanesia xanthocarpa (Mart.) O. Berg seedlings

Abstract The objective of this study was to evaluate the activity of antioxidant enzymes, the functioning of the photosystem II and quality of C. xanthocarpa seedlings cultivated under intermittent water deficit and shading levels and the influence of shading on recovery potential after suspension of the stress conditions. The seedlings were subjected to three levels of shading (0, 30, and 70%), six periods of evaluation (start: 0 days; 1st and 2nd photosynthesis zero: 1st and 2nd P0; 1st and 2nd recovery: 1stand 2nd REC; and END), and two forms of irrigation (control: periodically irrigated to maintain 70% substrate water retention capacity, and intermittent irrigation: suspension of irrigation). The plants subjected to intermittent irrigation conditions at 0% shading showed a reduction in water potential (w) and potential quantum efficiency of photosystem II (Fv/Fm) and maximum efficiency of the photochemical process (Fv/F0) and an increase in basal quantum production of the non-photochemical processes (F0/Fm). Superoxide dismutase (SOD) activity was higher in the leaves than in the roots. The C. xanthocarpa is a species sensitive to water deficit but presents strategies to adapt to an environment under temporary water restriction, which are more temporary are most efficient under shading. The seedlings with water deficit at all levels of shading exhibited higher protective antioxidant activity and lower quality at 0% shading. The shading minimizes prevents permanent damage to the photosystem II and after the re-irrigation, the evaluated characteristics showed recovery with respect to the control group, except POD and SOD activities in the leaves.

Resumo O objetivo deste estudo foi avaliar a atividade de enzimas antioxidantes, o funcionamento do fotossistema II e a qualidade de mudas de C. xanthocarpa cultivadas sob déficit hídrico intermitente e níveis de sombreamento e a influência do sombreamento sobre o potencial de recuperação após suspensão das condições de estresse. As mudas foram submetidas a três níveis de sombreamento (0, 30 e 70%), seis períodos de avaliação (início: 0 dias; 1ª e 2ª fotossíntese zero: 1ª e 2ª P0; 1ª e 2ª recuperação: 1ª e 2ª REC; e final), e duas formas de irrigação (controle: periodicamente irrigado para manter 70% da capacidade de retenção de água do substrato, e irrigação intermitente: suspensão da irrigação). As plantas submetidas às condições de irrigação intermitente a 0% de sombreamento apresentaram redução do potencial hídrico (w) e eficiência quântica potencial do fotossistema II (Fv/Fm) e máxima eficiência do processo fotoquímico (Fv/F0) e aumento da produção quantica basal dos processos não fotoquímicos (F0/Fm). A atividade da superóxido dismutase (SOD) foi maior nas folhas do que nas raízes. C. xanthocarpa é uma espécie sensível ao déficit hídrico, mas apresenta estratégias para se adaptar a um ambiente com restrição hídrica temporária, sendo mais eficientes sob sombreamento. As mudas com déficit hídrico em todos os níveis de sombreamento exibiram maior atividade antioxidante protetora e menor qualidade no sombreamento 0%. O sombreamento minimiza danos permanentes ao fotossistema II e após a re-irrigação, as características avaliadas apresentaram recuperação em relação ao grupo controle, exceto atividades de POD e SOD nas folhas.

Study of resveratrol against bone loss by using in-silico and in-vitro methods

Abstract By applying the in-silico method, resveratrol was docked on those proteins which are responsible for bone loss. The Molecular docking data between the resveratrol and Receptor activator of nuclear factor-kappa- ligand [RANKL] receptors proved that resveratrol binds tightly to the receptors, showed the highest binding affinities of 6.9, 7.6, 7.1, 6.9, 6.7, and 7.1 kcal/mol. According to in-vitro data, Resveratrol reduced the osteoclasts after treating Marrow-Derived Macrophages [BMM] with Macrophage colony-stimulating factor [MCSF] 20ng / ml and RANKL 50ng / ml, with different concentrations of resveratrol (2.5, 10 g / ml) For 7 days, the cells were treated with MCSF (20 ng / ml) and RANKL (40 ng / ml) together with concentrated trimethyl ether and resveratrol (2.5, 10 g / ml) within 12 hours. Which, not affect cell survival. After fixing osteoclast cells with formaldehyde fixative on glass coverslip followed by incubation with 0.1% Triton X-100 in PBS for 5 min and after that stain with rhodamine phalloidin staining for actin and Hoechst for nuclei. Fluorescence microscopy was performed to see the distribution of filaments actin [F.actin]. Finally, resveratrol reduced the actin ring formation. Resveratrol is the best bioactive compound for drug preparation against bone loss.

Resumo Com a aplicação do método in-silico, o resveratrol foi ancorado nas proteínas responsáveis pela perda óssea. Os dados de docking molecular entre o resveratrol e o ligante do receptor ativador do fator nuclear kappa- [Receptor Activator of Nuclear Factor kappa-B Ligant (RANKL)] provaram que o resveratrol se liga fortemente aos receptores, mostraram as afinidades de ligação mais altas de 6,9, 7,6, 7,1, 6,9, - 6,7 e -7,1 kcal / mol. De acordo com dados in-vitro, o resveratrol reduziu os osteoclastos após o tratamento de macrófagos derivados da medula óssea [Bone Marrow-derived Macrophage (BMM)] com fator estimulador de colônias de macrófagos [Macrophage Colony-Stimulating Factor (MCSF)] 20ng / ml e RANKL 50ng / ml, com diferentes concentrações de resveratrol (2,5, 10 g / ml). Durante sete dias, as células foram tratadas com MCSF (20 ng / ml) e RANKL (40 ng / ml) juntamente com éter trimetílico concentrado e resveratrol (2,5, 10 g / ml) em 12 horas, processo que não afeta a sobrevivência celular. Após a fixação de células de osteoclastos com fixador de formaldeído em lamela de vidro seguido de incubação com 0,1% Triton X-100 em PBS por 5 min, foi realizado posteriormente o procedimento para corar com rodamina faloidina a actina e Hoechst os núcleos. A microscopia de fluorescência foi realizada para ver a distribuição dos filamentos de actina [F.actina]. Finalmente, o resveratrol reduziu a formação do anel de actina. O resveratrol é o melhor composto bioativo para o preparo de medicamentos contra a perda óssea.

Shading minimizes the effects of water deficit in Campomanesia xanthocarpa (Mart) O Berg seedlings / Sombreamento minimiza o efeito do déficit hídrico em mudas de Campomanesia xanthocarpa (Mart) O Berg

The objective of this study was to evaluate the activity of antioxidant enzymes, the functioning of the photosystem II and quality of C. xanthocarpa seedlings cultivated under intermittent water deficit and shading levels and the influence of shading on recovery potential after suspension of the stress conditions. The seedlings were subjected to three levels of shading (0, 30, and 70%), six periods of evaluation (start: 0 days; 1st and 2nd photosynthesis zero: 1st and 2nd P0; 1st and 2nd recovery: 1stand 2nd REC; and END), and two forms of irrigation (control: periodically irrigated to maintain 70% substrate water retention capacity, and intermittent irrigation: suspension of irrigation). The plants subjected to intermittent irrigation conditions at 0% shading showed a reduction in water potential (Ψw) and potential quantum efficiency of photosystem II (Fv/Fm) and maximum efficiency of the photochemical process (Fv/F0) and an increase in basal quantum production of the non-photochemical processes (F0/Fm). Superoxide dismutase (SOD) activity was higher in the leaves than in the roots. The C. xanthocarpa is a species sensitive to water deficit but presents strategies to adapt to an environment under temporary water restriction, which are more temporary are most efficient under shading. The seedlings with water deficit at all levels of shading exhibited higher protective antioxidant activity and lower quality at 0% shading. The shading minimizes prevents permanent damage to the photosystem II and after the re-irrigation, the evaluated characteristics showed recovery with respect to the control group, except POD and SOD activities in the leaves.

O objetivo deste estudo foi avaliar a atividade de enzimas antioxidantes, o funcionamento do fotossistema II e a qualidade de mudas de C. xanthocarpa cultivadas sob déficit hídrico intermitente e níveis de sombreamento e a influência do sombreamento sobre o potencial de recuperação após suspensão das condições de estresse. As mudas foram submetidas a três níveis de sombreamento (0, 30 e 70%), seis períodos de avaliação (início: 0 dias; 1ª e 2ª fotossíntese zero: 1ª e 2ª P0; 1ª e 2ª recuperação: 1ª e 2ª REC; e final), e duas formas de irrigação (controle: periodicamente irrigado para manter 70% da capacidade de retenção de água do substrato, e irrigação intermitente: suspensão da irrigação). As plantas submetidas às condições de irrigação intermitente a 0% de sombreamento apresentaram redução do potencial hídrico (Ψw) e eficiência quântica potencial do fotossistema II (Fv/Fm) e máxima eficiência do processo fotoquímico (Fv/F0) e aumento da produção quantica basal dos processos não fotoquímicos (F0/Fm). A atividade da superóxido dismutase (SOD) foi maior nas folhas do que nas raízes. C. xanthocarpa é uma espécie sensível ao déficit hídrico, mas apresenta estratégias para se adaptar a um ambiente com restrição hídrica temporária, sendo mais eficientes sob sombreamento. As mudas com déficit hídrico em todos os níveis de sombreamento exibiram maior atividade antioxidante protetora e menor qualidade no sombreamento 0%. O sombreamento minimiza danos permanentes ao fotossistema II e após a re-irrigação, as características avaliadas apresentaram recuperação em relação ao grupo controle, exceto atividades de POD e SOD nas folhas.

Shading minimizes the effects of water deficit in Campomanesia xanthocarpa (Mart.) O. Berg seedlings / Sombreamento minimiza o efeito do déficit hídrico em mudas de Campomanesia xanthocarpa (Mart.) O. Berg

Abstract The objective of this study was to evaluate the activity of antioxidant enzymes, the functioning of the photosystem II and quality of C. xanthocarpa seedlings cultivated under intermittent water deficit and shading levels and the influence of shading on recovery potential after suspension of the stress conditions. The seedlings were subjected to three levels of shading (0, 30, and 70%), six periods of evaluation (start: 0 days; 1st and 2nd photosynthesis zero: 1st and 2nd P0; 1st and 2nd recovery: 1stand 2nd REC; and END), and two forms of irrigation (control: periodically irrigated to maintain 70% substrate water retention capacity, and intermittent irrigation: suspension of irrigation). The plants subjected to intermittent irrigation conditions at 0% shading showed a reduction in water potential (Ψw) and potential quantum efficiency of photosystem II (Fv/Fm) and maximum efficiency of the photochemical process (Fv/F0) and an increase in basal quantum production of the non-photochemical processes (F0/Fm). Superoxide dismutase (SOD) activity was higher in the leaves than in the roots. The C. xanthocarpa is a species sensitive to water deficit but presents strategies to adapt to an environment under temporary water restriction, which are more temporary are most efficient under shading. The seedlings with water deficit at all levels of shading exhibited higher protective antioxidant activity and lower quality at 0% shading. The shading minimizes prevents permanent damage to the photosystem II and after the re-irrigation, the evaluated characteristics showed recovery with respect to the control group, except POD and SOD activities in the leaves.

Resumo O objetivo deste estudo foi avaliar a atividade de enzimas antioxidantes, o funcionamento do fotossistema II e a qualidade de mudas de C. xanthocarpa cultivadas sob déficit hídrico intermitente e níveis de sombreamento e a influência do sombreamento sobre o potencial de recuperação após suspensão das condições de estresse. As mudas foram submetidas a três níveis de sombreamento (0, 30 e 70%), seis períodos de avaliação (início: 0 dias; 1ª e 2ª fotossíntese zero: 1ª e 2ª P0; 1ª e 2ª recuperação: 1ª e 2ª REC; e final), e duas formas de irrigação (controle: periodicamente irrigado para manter 70% da capacidade de retenção de água do substrato, e irrigação intermitente: suspensão da irrigação). As plantas submetidas às condições de irrigação intermitente a 0% de sombreamento apresentaram redução do potencial hídrico (Ψw) e eficiência quântica potencial do fotossistema II (Fv/Fm) e máxima eficiência do processo fotoquímico (Fv/F0) e aumento da produção quantica basal dos processos não fotoquímicos (F0/Fm). A atividade da superóxido dismutase (SOD) foi maior nas folhas do que nas raízes. C. xanthocarpa é uma espécie sensível ao déficit hídrico, mas apresenta estratégias para se adaptar a um ambiente com restrição hídrica temporária, sendo mais eficientes sob sombreamento. As mudas com déficit hídrico em todos os níveis de sombreamento exibiram maior atividade antioxidante protetora e menor qualidade no sombreamento 0%. O sombreamento minimiza danos permanentes ao fotossistema II e após a re-irrigação, as características avaliadas apresentaram recuperação em relação ao grupo controle, exceto atividades de POD e SOD nas folhas.

Study of resveratrol against bone loss by using in-silico and in-vitro methods / Estudo do resveratrol contra a perda óssea usando métodos in silico e in vitro

Abstract By applying the in-silico method, resveratrol was docked on those proteins which are responsible for bone loss. The Molecular docking data between the resveratrol and Receptor activator of nuclear factor-kappa-Β ligand [RANKL] receptors proved that resveratrol binds tightly to the receptors, showed the highest binding affinities of −6.9, −7.6, −7.1, −6.9, −6.7, and −7.1 kcal/mol. According to in-vitro data, Resveratrol reduced the osteoclasts after treating Marrow-Derived Macrophages [BMM] with Macrophage colony-stimulating factor [MCSF] 20ng / ml and RANKL 50ng / ml, with different concentrations of resveratrol (2.5, 10 μg / ml) For 7 days, the cells were treated with MCSF (20 ng / ml) and RANKL (40 ng / ml) together with concentrated trimethyl ether and resveratrol (2.5, 10 μg / ml) within 12 hours. Which, not affect cell survival. After fixing osteoclast cells with formaldehyde fixative on glass coverslip followed by incubation with 0.1% Triton X-100 in PBS for 5 min and after that stain with rhodamine phalloidin staining for actin and Hoechst for nuclei. Fluorescence microscopy was performed to see the distribution of filaments actin [F.actin]. Finally, resveratrol reduced the actin ring formation. Resveratrol is the best bioactive compound for drug preparation against bone loss.

Resumo Com a aplicação do método in-silico, o resveratrol foi ancorado nas proteínas responsáveis ​​pela perda óssea. Os dados de docking molecular entre o resveratrol e o ligante do receptor ativador do fator nuclear kappa-Β [Receptor Activator of Nuclear Factor kappa-B Ligant (RANKL)] provaram que o resveratrol se liga fortemente aos receptores, mostraram as afinidades de ligação mais altas de −6,9, −7,6, −7,1, −6,9, - 6,7 e -7,1 kcal / mol. De acordo com dados in-vitro, o resveratrol reduziu os osteoclastos após o tratamento de macrófagos derivados da medula óssea [Bone Marrow-derived Macrophage (BMM)] com fator estimulador de colônias de macrófagos [Macrophage Colony-Stimulating Factor (MCSF)] 20ng / ml e RANKL 50ng / ml, com diferentes concentrações de resveratrol (2,5, 10 μg / ml). Durante sete dias, as células foram tratadas com MCSF (20 ng / ml) e RANKL (40 ng / ml) juntamente com éter trimetílico concentrado e resveratrol (2,5, 10 μg / ml) em 12 horas, processo que não afeta a sobrevivência celular. Após a fixação de células de osteoclastos com fixador de formaldeído em lamela de vidro seguido de incubação com 0,1% Triton X-100 em PBS por 5 min, foi realizado posteriormente o procedimento para corar com rodamina faloidina a actina e Hoechst os núcleos. A microscopia de fluorescência foi realizada para ver a distribuição dos filamentos de actina [F.actina]. Finalmente, o resveratrol reduziu a formação do anel de actina. O resveratrol é o melhor composto bioativo para o preparo de medicamentos contra a perda óssea.

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.

Development and application of a method for determination of activity and vitality of Hansenula polymorpha by carboxyfluorescein diacetate-propidium iodide dua fluorescent staining / 中国生物制品学杂志

@#Objective To develop a method for determination of activity and viability of Hansenula polymorpha by dual fluorescent staining with carboxyfluorescein diacetate(CFDA)and propidium iodide(PI).Methods The time durations(5,10,20,40,60 and 120 min for CFDA,5,10,15,20 and 25 min for PI)and working solution concentrations(50,100,200and 300 μg for CFDA,2,10,20 and 30 μmol/L for PI)for the dual fluorescent staining were optimized by single factor test. Under the optimal condition,the H.polymorpha samples at theoretical survival rates of 0,25%,50%,75% and 100%were determined,of which the fluorescent intensity was observed under fluorescent microscope,and the gray value was analyzed by ImageJ software. The live and dead cells were counted,based on which the actual survival and death rates were calculated. Meanwhile,the relationships of actual gray value,actual survival rate and actual death rate to the corresponding theoretical values were analyzed. Activity and viability of three batches of cultured H.polymorpha were detected by CFDA-PI dual fluorescence staining.Results The optimal time durations for staining with CFDA and PI were 60 and 5 min,while the optimal working solution concentrations were 200 and 2 μmol/L,respectively. The actual gray value,actual survival rate and actual death rate of H.polymorpha samples at various theoretical survival rates were significantly correlated to the corresponding theoretical values(R~2=0. 998 3～0. 999 2,P < 0. 05). The CVs of activity and viability values in three detections of three batches of H.polymorpha culture were 3. 20%～4. 03% and 1. 10%～2. 27%, respectively.Conclusion The CFDA-PI dual fluorescent staining was successfully developed,which may be used for determination of activity and vitality of H.polymorpha.

Optimization of expression conditions of dengue virus NS2B-NS3 protease in E.coli and determination of enzyme activity / 中国生物制品学杂志

@#Objective To express dengue virus(DENV)NS2B-NS3 protease in E.coli,optimize the expression conditions and determine the enzyme activity,so as to lay a foundation of screening and discovering of lead compounds targeting DENV.Methods Codon-optimized NS2B-NS3 gene was inserted into pET-28a vector to construct recombinant prokaryotic expression plasmid pET-28a-NS2B-NS3,which was transformed E.coli Rosetta(DE3)competent cells and induced by IPTG to express NS2B-NS3 protease. The optimal expression conditions of NS2B-NS3 protease in E.coli were determined by optimizing induction length,induction temperature and IPTG concentration. NS2B-NS3 protease was isolated and purified by HisTrap~(TM) affinity chromatography column and measured for the protease activity by fluorescence resonance energy transfer(FRET)assay.Results The recombinant prokaryotic expression plasmid pET-28a-NS2B-NS3 was constructed correctly as identified by restriction analysis(NheⅠ/XhoⅠ)and sequencing. The optimal expression conditions of NS2BNS3 protease in E.coli were as follows:induction temperature of 20 ℃,induction length of 10 h and IPTG concentration of0. 2 mmol/L. The purified NS2B-NS3 protease showed a purity of more than 90% with a exhibited a of 20 mg/L,which bound to mouse monoclonal antibody against His-tag specifically and had good hydrolytic activity with a specific activity of 16. 111 U/mg,a K_m of 16. 46 μmol/L and a k_(cat) of 0. 028/s.Conclusion DENV NS2B-NS3 protease with high purity and activity was successfully prepared,which laid an experimental foundation of the establishment of high-throughput screening model for inhibitors targeting NS2B-NS3 protease.

Correlation between in-vivo mercury exposure from Cinnabaris and memory disorders in juvenile rat / 中国药科大学学报

@#Cinnabaris(α-HgS) is a mineral traditional Chinese material medica, as a tranquilizer and sedative, which is widely used in combination with herbs for the treatment of children high fever and convulsion.However, a large amount of mercury in Cinnabaris poses a potential risk to the immature central nervous system of children and probably causes severe memory disorders.Inthisstudy,three groups of juvenile rats were given low, medium, and high doses of Cinnabaris by oral gavage once a day for 14 continuous weeks, respectively.The blood mercury concentrations of the rats at different growth phases were monitored by atomic fluorescence spectrometry.The brain structural and functional changes related to the memory functions were investigated through HE staining and Morris water-maze test. Correlation analysis was conducted to clarify the dose- mercury exposure-toxic effect relationship of Cinnabaris and memory disorders.It was found thatthe blood mercury levels increased in both time- and dose-dependent manner.After the 14-week continuous administration of Cinnabaris, the pathological lesions in hippocampal neurons of rats in the high dose group were observed including pyknosis and disordered cell arrangement.In the Morris water-maze test, compared with the control group, rats in the high dose group exhibited the significantly prolonged latency to find the platform and the target quadrant, and the time spent in the target quadrant was obviously shortened. Thus, the significant correlations were established between Cinnabaris dose and mercury exposure,mercury exposure and memory disorders, respectively. In conclusion, the long-term and overdose administration of Cinnabaris in juvenile rats can increase the in-vivo mercury level, destroy the normal hippocampal morphological structure, and lead to memory disorders. This study provided scientific references for the potential mercury poisoning risks pharmacovigilance of Cinnabaris-containing paediatric formulations.

Research Progress of Indocyanine Green Fluorescence Laparoscopic Technique in Clinical Application / 中国医疗器械杂志

Indocyanine green (ICG) is the most commonly used near-infrared fluorescent (NIRF) dye in clinical practice, and its mediated near-infrared fluorescence imaging technology is gradually applied in clinical practice. It has shown great potential in invasive surgery (MIS) and is expected to become the standard technology for surgical diagnosis and treatment of diseases. The clinical application of ICG fluorescence laparoscopy is reviewed here.