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.

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.

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.

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.

Effect of fluorescence method on cardiopulmonary function, immune function and safety during pulmonary segmental resection / 中国医师进修杂志

Objective:To explore the effects of fluorescence method and improved expansion and collapse method on cardiopulmonary function, immune function and safety during segmental pneumonectomy.Methods:One hundred and fifty-six patients with lung segmental resection operation in Yucheng People′s Hospital from March 2018 to August 2020 were enrolled. They were divided into two groups according to the random number table method, each group with 78 patients. The patients in the observation group were treated with fluorescence method and the patients in the control group were treated with modified expansion collapse method. The operation indexes, immune function and cardiopulmonary function of the two groups were compared, and the incidence of postoperative complications were counted.Results:The intraoperative time and the appearance time of intersegmental plane in the observation group were shorter than those in the control group: (105.33 ± 10.62) min vs. (139.46 ± 12.58) min, (15.46 ± 1.13) s vs. (724.56 ± 65.65) s, there were statistical differences ( P<0.05). After operative for 1, 7 d, the levels of 6 min walking distance, peak expiratory flow rate, percentage of forced vital capacity and percentage forced expiratory volume in 1 second in the observation group were higher than those in the control group: after operative for 1 d: (475.36 ± 10.29) m vs. (412.11 ± 13.33) m, (284.52 ± 10.33) L/min vs. (251.13 ± 12.89) L/min, (85.65 ± 3.21)% vs. (81.13 ± 1.43)%, (83.25 ± 2.55)% vs. (74.49 ± 2.53)%; after operative for 7 d: (510.23 ± 16.66) m vs. (488.33 ± 15.42) m, (302.13 ± 15.58) L/min vs. (285.12 ± 10.22) L/min, (93.46 ± 5.79)% vs. (88.44 ± 5.44)%, (92.25 ± 2.32)% vs. (85.54 ± 2.13)%, there were statistical differences ( P<0.05). After operative for 1, 7 d, the levels of CD 4+/CD 8+, CD 8+, CD 4+ in the observation group were higher than those in the control group: after operative for 1 d: 0.85 ± 0.10 vs. 0.52 ± 0.04, 0.305 ± 0.025 vs. 0.285 ± 0.012, 0.325 ± 0.021 vs. 0.304 ± 0.025; after operative for 7 d: 1.13 ± 0.10 vs. 1.02 ± 0.07, 0.324 ± 0.029 vs. 0.306 ± 0.023, 0.359 ± 0.024 vs. 0.332 ± 0.025, there were statistical differences ( P<0.05). The rate of postoperative complications in the observation group was lower than that in the control group: 2.56%(2/78) vs. 11.54%(9/78), there was statistical difference ( χ2 = 4.79; P = 0.029). Conclusions:Both the fluorescence method and the modified inflation-collapse method have certain therapeutic effects, but the combination of fluorescence method in segmentectomy can provide reliable technical support and reduce the impact on cardiopulmonary function.

Scientific and technological innovation drives the development of minimally invasive surgery / 中华消化外科杂志

Scientific and technological innovation is the most important role in driving the development of minimally invasive surgery. After more than 30 years of development, modern mini-mally invasive surgery represented by laparoscopic surgery has gradually matured. Various types of minimally invasive surgeries have been popularized, and the difficulty of surgery has changed from extreme to limit. Surgical equipments and instruments can meet the needs of most clinical operations. The future of minimally invasive surgery has reached a crossroad, and only scientific and technological innovation can promote the development of minimally invasive surgery change lanes and overtake, ushering in new development, new methods, and a new world. For innovation, the most important thing is not knowledge, but vision and ideas. A new technological revolution will inevitably bring about changes in the industry. What changes will be ushered in the operation and treatment of diseases in the future? What will be the breakthrough of minimally invasive surgery? It is worth to wait and see. The authors discuss the development direction of minimally invasive surgery based on the recent application of hot technologies in laparoscopic surgery.

Progress of research and technical in minimally invasive surgery for gastric cancer / 中华消化外科杂志

Gastric cancer is one of the most common malignant tumors in the world, and its incidence and mortality are among the top of malignant tumors in China. Since Kitano et al com-plete the first laparoscopic radical gastrectomy for gastric cancer in the world in 1992, the laparos-copic technology has developed rapidly. After more than 30 years of exploration and practice, the clinical diagnosis and treatment of gastric cancer in China has also made considerable progress. A large number of clinical studies at home and abroad have confirmed that laparoscopic radical gas-trectomy is no less effective than traditional open surgery in the short and long term. Laparoscopic radical gastrectomy has the characteristics of less trauma, faster recovery of gastrointestinal func-tion, less postoperative pain, and shorter average hospital stay. It has gradually replaced open surgery as the mainstream surgical method for gastric cancer. As the concept of surgical treatment for gastric cancer continues to update, emerging minimally invasive technologies continue to emerge, including robotic surgery systems and indocyanine green tracing technology, which are increasingly used in gastric cancer surgery, making gastric cancer surgery more minimally invasive and accurate, the quality of perigastric lymph node dissection and the domestic gastric cancer surgery technology further improving. Based on the relevant research at home and abroad, the authors review and summarize the latest progress in recent years with the topic of minimally invasive surgery for gastric cancer, aiming to systematically describe the current situation and future prospects of gastric cancer surgery. It is believed that in the future, the clinical diagnosis and treatment of gastric cancer in China will be more standardized, minimally invasive and accurate, more high-quality multicenter clinical research will be carry out and the diagnosis and treatment of gastric cancer will be further improved in China.

Research advances in near-infrared fluorescence imaging technology for breast cancer / 国际外科学杂志

Near-infrared fluorescence imaging (NIRFI) is a new noninvasive detection and diagnosis technology, with the continuous development of NIRFI technology, now widely used in the clinic, characterized by high sensitivity, high penetration, no harmful radiation and simple equipment operation. This article describes the recent applications of NIRFI in the diagnosis and treatment of breast cancer and looks at future developments and perspectives in this field.

Palmitoylation in Regulating Fyn Kinase Activity Based on FRET Imaging / 医用生物力学

Objective To investigate the molecular mechanism of palmitoylation modification in regulating the activity of non-receptor tyrosine kinase Fyn. Methods The intracellular Fyn activity was detected by applying fluorescence resonance energy transfer (FRET) technology, and the mechanism was investigated by combining with Fyn palmitoylation deficiency and C-terminal Src kinase ( CSK ) plasmid co-expression. ResultsExperimental data showed that single loss of either of ( C3, C6) palmitoylation sites resulted in higher Fyn activity, and C6 seemed more significant. It is known that CSK membrane translocation occurred after activation. FRET assay confirmed that CSK could down-regulate the activity of Fyn in cells, but could not effectively regulate the activity of Fyn(GSS) with the loss of palmitoylation sites. Conclusions The results in this study support the hypothesis on Fyn regulation by spatial localization, namely, non-palmitoylated Fyn (GSS) is less effective in the inhibitory regulation by CSK on cell membrane, thus promoting constitutive high activity expression

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.

Clinical value of fluorescence in situ hybridization with MDM2 and DDIT3 probe in diagnosis of liposarcoma / 北京大学学报(医学版)

OBJECTIVE@#To investigate the value of using MDM2 amplification probe and DDIT3 dual-color, break-apart rearrangement probe fluorescence in situ hybridization (FISH) technique in the diagnosis of liposarcoma.@*METHODS@#In the study, 62 cases of liposarcoma diagnosed in Peking University First Hospital from January 2015 to December 2019 were analysed for clinicopathological information. Of these 62 cases of liposarcoma, all were analysed for MDM2 amplification and 48 cases were analysed for DDIT3 rearrangement using a FISH technique. Our study aimed to evaluate the status of MDM2 and DDIT3 by FISH in liposarcoma and correlate it with diagnosis of different subtypes of liposarcoma. The subtypes of liposarcoma were classified according to the FISH results, combined with the relevant clinicopathological features.@*RESULTS@#The patients aged 31-89 years (mean: 59 years) with a 1.75:1 male to female ratio. Histologically, there were 20 cases of atypical lipomatous tumour/well-differentiated liposarcoma (ALT/WDLPS), 26 cases of dedifferentiated liposarcoma (DDLPS), 13 myxoid liposarcoma (MLPS) and 3 pleomorphic liposarcoma (PLPS). Tumors with DDLPS (23/26) and WDLPS (8/20) were localized retroperitoneally, while both tumours of MLPS and PLPS were localized extra-retroperitoneally, and the difference of sites among the four subtypes of liposarcoma was statistically significant (P < 0.05). Histologically, varied mucoid matrix could be observed in the four subtypes of liposarcoma, and the difference was statistically significant (P < 0.05). MDM2 gene amplification was demonstrated in all cases of ALT/WDLPS and DDLPS (100%, 20/20 and 26/26 respectively); DDIT3 gene rearrangement was noted only in MLPS (100%, 13/13); most cases of DDLPS (96.2%, 25/26) and ALT/WDLPS (83.3%, 5/6, 6 cases selected for detection) demonstrated the picture of amplification of the DDIT3 telomeric tag. According to the instructions of DDIT3 break-apart rearrangement probe, the 5' telomere probe and 3' centromere probe spanned but did not cover the DDIT3 gene itself, on the contrary, the 5' telomere probe covered the CDK4 gene, while the DDIT3 and CDK4 gene were located adjacent to each other on chromosome, therefore, when the amplification signal appeared on the telomeric tag of the DDIT3 rearrangement probe, it indeed indicated the CDK4 gene amplification rather than the DDIT3 gene rearrangement. Then the 10 cases with DDIT3 telomeric tag amplification were selected for CDK4 and DDIT3 gene amplification probe FISH tests, and all the cases showed CDK4 gene amplification (100%, 10/10) and two of the 10 cases demonstrated co-amplification of CDK4 and DDIT3 (20%, 2/10); DDIT3 polysomy detected by DDIT3 gene rearrangement probe was found in 1 case of DDLPS and 2 cases of PLPS (66.7%, 2/3) with morphology of high-grade malignant tumour and poor prognosis.@*CONCLUSION@#Our results indicate that a diagnosis of different subtype liposarcoma could be confirmed based on the application of MDM2 and DDIT3 FISH, combined with clinicopathological findings. It is also noteworthy that atypical signals should be correctly interpreted to guide correct treatment of liposarcomas.

Application of indocyanine green in near-infrared fluorescence imaging to detect necrotic bone associated with osteoradionecrosis of the jaws / 口腔疾病防治

Objective @# To investigate the application of indocyanine green in near-infrared fluorescence imaging to determine the scope of necrotic bone resection in osteoradionecrosis of the jaw and to provide a reference for clinicians@*Methods @#Eight patients with osteoradionecrosis of the jaws were enrolled. Indocyanine green was intravenously injected through the elbow vein 10 minutes before osteotomy. After conservative resection of necrotic bone lesions based on imaging results, the scope of potential dead bone resection in the area of low fluorescence intensity was gradually expanded at an initial distance of 0.3 cm. Near-infrared fluorescence imaging and fluorescence intensity determination of bone cross-section were performed before and after extended resection. Statistical differences were analyzed. All patients with osteonecrosis underwent regular follow-up to evaluate the postoperative efficacy@*Results@#Indocyanine green was injected into all 8 patients with osteoradionecrosis for near-infrared fluorescence imaging and the scans were clear; the fluorescence intensity of fresh bone wounds with an expanded mandibular resection range of (0.95 ± 0.14) cm was (226.2 ± 15.8) au, which was higher than that based on intraoperative macroscopic observation and radiological results (108.8 ± 3.4) au, (t = 20.718, P<0.001). The postoperative follow-up improvement rate of 8 patients was 87.5%.@* Conclusion @#Near-infrared fluorescence imaging with indocyanine green can assist in the successful removal of necrotic bone until fresh bleeding of the jaw wound occurs, which has important clinical value in defining the resection range of osteoradionecrosis of the jaw.

Establishment and Application of Rapid Screening Method for Influenza Virus Neuraminidase Inhibitors Sourced from Traditional Chinese Medicine Based on Fluorescence Detection / 中国实验方剂学杂志

ObjectiveTo establish a rapid screening method for influenza virus neuraminidase（NA） inhibitors sourced from Chinese medicines based on fluorescence detection. MethodThe method was constructed based on the principle that after the reaction of the test sample and a certain amount of NA， the activity of some NA will be inhibited by the test sample， and the NA that is still active after the addition of the substrate can generate fluorescence at a specific wavelength when combined with the fluorescent substrate， and the inhibition rate of the test sample on NA was calculated according to the measured fluorescence intensity， so as to evaluate the in vitro inhibitory activity of the test sample on NA. A total of 49 high-purity chemical components from 12 Chinese medicines were used to evaluate the in vitro anti-NA activity by the established method. The theoretical calculated values of binding energy and inhibition constant after docking between the NA protein receptor and the test sample were used to prove the accuracy of the experimental results. The established method was applied to detect the in vitro NA inhibitory activity of different batches of Banlangen granules and Kangbingdu granules， so as to evaluate the quality consistency among different batches of samples. ResultThe methodological examination results showed that the method had good accuracy and repeatability. The screening results of 49 components showed that 22 of them had strong in vitro inhibitory activity against NA than peramivir ［half inhibitory concentration（IC50） was 131.2 μmol·L-1］， such as schaftoside， isoorientin， chebulinic acid， menthone and isoschaftoside. The inhibitory activity of the remaining 27 components was weaker than that of peramivir. The molecular docking results showed that the theoretical calculation results of binding energies and inhibition constants of most compounds were basically consistent with the experimental results. The test results of the inhibitory activity of 12 batches of Banlangen granules on NA showed that the quality consistency among samples A1， A2， B2， C1， C2， E2 and F2 was good. The analysis results of the inhibitory activity of 9 batches of Kangbingdu granules produced by the same manufacturer on NA showed that the inhibitory rates of samples K1 to K9 were 37.68%， 36.18%， 31.37%， 33.98%， 40.36%， 33.76%， 40.69%， 41.08%， 40.06% when the concentration of 0.02 g·mL-1， and the average inhibitory rate was 37.24%. ConclusionIn this paper， we successfully established an analytical method that can be used to rapidly evaluate whether Chinese medicines （derived from chemical components of traditional Chinese medicine or proprietary Chinese medicines） have in vitro anti-NA activity， which can be a powerful supplement to the existing screening methods for influenza virus NA inhibitors. And this method was used to screen 22 compounds from 12 Chinese medicines with good in vitro inhibitory activity against NA， which can provide candidate compounds for the development of anti-influenza small molecule drugs.

Advances in methods for detecting plastics biodegradation / 生物工程学报

The pollution caused by improper handling of plastics has become a global challenge. In addition to recycling plastics and using biodegradable plastics, an alternative solution is to seek efficient methods for degrading plastics. Among them, the methods of using biodegradable enzymes or microorganisms to treat plastics have attracted increasing attention because of its advantages of mild conditions and no secondary environmental pollution. Developing highly efficient depolymerizing microorganisms/enzymes is the core for plastics biodegradation. However, the current analysis and detection methods cannot meet the requirements for screening efficient plastics biodegraders. It is thus of great significance to develop rapid and accurate analysis methods for screening biodegraders and evaluating biodegradation efficiency. This review summarizes the recent application of various commonly used analytical techniques in plastics biodegradation, including high performance liquid chromatography, infrared spectroscopy, gel permeation chromatography, and determination of zone of clearance, with fluorescence analysis techniques highlighted. This review may facilitate standardizing the characterization and analysis of plastics biodegradation process and developing more efficient methods for screening plastics biodegraders.

Potential unreliability of ALK variant allele frequency in the efficacy prediction of targeted therapy in NSCLC / 医学前沿

Anaplastic lymphoma kinase (ALK) is the most common fusion gene involved in non-small cell lung cancer (NSCLC), and remarkable response has been achieved with the use of ALK tyrosine kinase inhibitors (ALK-TKIs). However, the clinical efficacy is highly variable. Pre-existing intratumoral heterogeneity (ITH) has been proven to contribute to the poor treatment response and the resistance to targeted therapies. In this work, we investigated whether the variant allele frequencies (VAFs) of ALK fusions can help assess ITH and predict targeted therapy efficacy. Through the application of next-generation sequencing (NGS), 7.2% (326/4548) of patients were detected to be ALK positive. On the basis of the adjusted VAF (adjVAF, VAF normalization for tumor purity) of four different threshold values (adjVAF < 50%, 40%, 30%, or 20%), the association of ALK subclonality with crizotinib efficacy was assessed. Nonetheless, no statistical association was observed between median progression-free survival (PFS) and ALK subclonality assessed by adjVAF, and a poor correlation of adjVAF with PFS was found among the 85 patients who received first-line crizotinib. Results suggest that the ALK VAF determined by hybrid capture-based NGS is probably unreliable for ITH assessment and targeted therapy efficacy prediction in NSCLC.

Treatment of rheumatoid arthritis by injection of sinomenine solid lipid nanoparticles under a fluorescence endoscopic laser confocal microscope / 中国中药杂志

A fluorescence endoscopic laser confocal microscope(FELCM) was used to direct the injection of sinomenine solid lipid nanoparticles(Sin-SLN) into the joint, and the in vitro effectiveness of Sin-SLN in the treatment of rheumatoid arthritis(RA) was evaluated. Sin-SLN was prepared with the emulsion evaporation-low temperature curing method. The Sin-SLN prepared under the optimal conditions showed the encapsulation efficiency of 64.79%±3.12%, the drug loading of 3.84%±0.28%, the average particle size of(215.27±4.21) nm, and the Zeta potential of(-32.67±0.84) mV. Moreover, the Sin-SLN demonstrated good stability after sto-rage for 30 days. The rabbit model of RA was established by the subcutaneous injection of ovalbumin and complete Freund's adjuvant. Five groups were designed, including a control group, a model group, a Sin(1.5 mg·kg~(-1)) group, a Sin-SLN(1.5 mg·kg~(-1)) group, and a dexamethasone(positive drug, 1.0 mg·kg~(-1), ig) group. The control group and the model group only received puncture treatment without drug injection. After drug administration, the local skin temperature and knee joint diameter were monitored every day. The knee joint diameter and the local skin temperature were lower in the drug administration groups than in the model group(P<0.05, P<0.01). FELCM recorded the morphological alterations of the cartilage of knee joint. The Sin-SLN group showed compact tissue structure and smooth surface of the cartilage. Enzyme-linked immunosorbent assay(ELISA) was employed to determine the serum le-vels of interleukin-1(IL-1) and tumor necrosis factor-α(TNF-α). The findings revealed that the Sin-SLN group had lower IL-1 and TNF-α levels than the model group(P<0.05, P<0.01). Hematoxylin-eosin(HE) staining was employed to reveal the pathological changes of the synovial tissue, which were significantly mitigated in the Sin-SLN group. The prepared Sin-SLN had uniform particle size and high stability. Through joint injection administration, a drug reservoir was formed. Sin-SLN effectively alleviate joint swelling and cartilage damage of rabbit, down-regulated the expression of inflammatory cytokines, and inhibited the epithelial proliferation and inflammatory cell infiltration of the synovial tissue, demonstrating the efficacy in treating RA.

Frontiers and development in live-cell super-resolution fluorescence microscopy / 生物医学工程学杂志

This paper reviews the research progress on live-cell super-resolution fluorescence microscopy, discusses the current research status and hotspots in this field, and summarizes the technological application of super-resolution fluorescence microscopy for live-cell imaging. To date, this field has gained progress in numerous aspects. Specifically, the structured illumination microscopy, stimulated emission depletion microscopy, and the recently introduced minimal photon fluxes microscopy are the current research hotspots. According to the current progress in this field, future development trend is likely to be largely driven by artificial intelligence as well as advances in fluorescent probes and relevant labelling methods.