ObjectiveTo compare the differences between the ultrafiltration method and the dextran gel filtration method during the purification of Tfn-modified PCL micelles by using purification efficiency and micelle purity as indicators. MethodsCoumarin-6 （C6） was used as a fluorescent probe and was loaded into HOOC-PEG-PCL to form PCL micelles by the film-dispersion method. Tfn was then conjugated to the surface of PCL micelles via an amidation reaction， resulting in two types of micelles： Tfn/PCL_H and Tfn/PCL_L. The pharmaceutical properties of the two types of micelles were characterized. The micelles were then purified through ultrafiltration method and dextran gel method respectively， and the efficiency of the two methods， along with the purity of the final micelles， was compared. The density of Tfn on the surface of PCL micelles was also calculated. ResultsThe hydrated diameter of PCL micelles was approximately 73 nm， and the C6 loading efficiency was around 0.046%. The size increased to 134 nm and 158 nm for Tfn/PCL_L and Tfn/PCL_H， respectively. The micelle population was monodisperse. The purification results showed that， for the ultrafiltration method， after two and one rounds of purification， the Tfn/C6 ratio stabilized at 23.6 and 3.4 for Tfn/PCL_H and Tfn/PCL_L， respectively. For the dextran gel filtration method， the Tfn/C6 ratio reached 23.7 for the Tfn/PCL_H group after two rounds of purification. However， for the Tfn/PCL_L group， the Tfn/C6 ratio increased during four rounds of dextran gel purification， and a significant difference （P = 0.042 4） was observed between the first and last filtrations. The density of Tfn in the final micelles were calculated. For the ultrafiltration method， the Tfn density of Tfn/PCL_H and Tfn/PCL_L were 94.9% and 13.8%， respectively. For the dextran gel filtration method， the density of the two micelles were 95.6% and 14.4%， respectively. For Tfn/PCL_L group， the density results revealing a statistically significant difference （P=0.000 2）. ConclusionThe purification efficiency of the two methods is comparable. However， the purity of the final micelles shows a significant difference， with the dextran gel filtration method resulting in higher purity， particularly for the Tfn/PCL_L micelles.

The official implementation of the Regulations on Clinical Research and Translation of New Biomedical Technologies (State Council Decree No.818, hereinafter referred to as the "Regulations") marks a new stage of legal governance, standardization, and whole-chain regulation for emerging biomedical fields such as cell and gene therapy (CGT) in China, exerting a profound impact on the development of transfusion medicine as a discipline. Within the framework of the core regulatory systems established by the Regulations, and drawing upon the extensive experience of transfusion medicine in comprehensive quality control, cell processing and handling, ethical compliance, and clinical translation, this study systematically elucidates the intrinsic alignment between transfusion medicine and CGT with respect to technical workflows, quality control rationale, and management systems. It further clarifies the discipline-specific advantages of the department of transfusion medicine to proactively undertake CGT engagement, while analyzing current shortcomings and transformational bottlenecks. By leveraging the existing institutional framework, the department of transfusion medicine is well positioned to evolve into a core platform for whole-process quality control and a pivotal hub for in-hospital clinical translation of CGT. However, this transformation must concurrently address multiple challenges, including heightened compliance responsibilities, lagging technical infrastructure, a shortage of interdisciplinary talent, and increasingly stringent standards for clinical research and translation. The Regulations provide a legal basis and policy support for the department of transfusion medicine to transcend its traditional, singular function of blood safety assurance and establish a synergistic development model integrating blood safety and cell therapy. Therefore, the department of transfusion medicine should proactively align with regulatory requirements, refine whole-process compliance systems, upgrade technical platforms, and cultivate interdisciplinary professionals to achieve high-quality transformative development as a core supporting discipline for biotherapy.

Objective:In order to reduce the trauma associated with fetal cardiopulmonary bypass(F-CPB), Our team plans to develop a minimally invasive F-CPB through a small incision in the right axilla. The efficacy of this technique will be verified by using a big experimental animal model, thereby laying the foundation for fetal cardiac surgery supported by F-CPB in the future.Methods:Ten pregnant sheep were divided into F-CPB group(n=5) and control group(n=5). After fasting for 24 h, fetal lambs in the F-CPB group underwent a right axillary incision to establish F-CPB running for 1 h; The control group of fetal lambs only expose heart 1 h without F-CPB. Collect blood sample for laboratory test at the CPB vehicle before(T0), 30 min(T1), and 1 h after F-CPB running(T2) for the F-CPB group and through Superior Vena Cava before(T0), 30 min(T1), and 1 h after F-CPB running(T2) for the control group.Results:The blood routine indicators such as RBC, HCT, and Hb in the F-CPB group of fetal lambs decreased significantly during F-CPB, and their distribution showed significant statistical differences compared with the control group( P<0.05). There were no significant statistical differences in blood gas indicators such as pH, PO 2, PCO 2, and lactate concentration between the F-CPB group and the control group( P>0.05). There was no statistically significant difference in the concentration of cTnI in fetal lamb serum at each time point( P>0.05). There were significant statistical differences( P<0.05) in the distribution of fetal lamb Alb, γ-GGT, CK concentration and cholesterol concentration at various time points in the F-CPB group compared with the control group in liver function examination. In addition, the distribution of BUN in fetal lambs showed a significant difference between the two groups( P=0.006). Conclusion:A minimally invasive F-CPB via small incision in the right axilla is safe and feasible. The experimental animal model has demonstrated that this technique has minimal impact on the vital organ functions and internal environment of fetal lambs, thereby laying the foundation for clinical fetal cardiac surgery in the future.

Melatonin is widely used as an over-the-counter medication to treat insomnia in children with autism spectrum disorder (ASD) and neurogenetic disorders (NGD). However, there is still a lack of research on its efficacy and safety, and clinical practice standards are to be established. In response, the International Pediatric Sleep Association (IPSA) convened an expert panel and developed a consensus statement:"Melatonin Use in Managing Insomnia in Children with Autism and Other Neurogenetic Disorders-an Assessment by the International Pediatric Sleep Association (IPSA)", which was published in Sleep Medicine, April 2024. The consensus focused on the efficacy and adverse effects of melatonin treatment for insomnia in children with ASD and NGD-including Smith-Magenis syndrome, Rett syndrome, Angelman syndrome, and tuberous sclerosis complex. It systematically reviews randomized controlled trials (RCTs) conducted between 2012 and 2022, and integrates current best clinical practices to formulate 10 consensus recommendations. Despite these contributions, the consensus has limitations: a small number of included RCTs, a lack of grading for evidence quality, and recommendation strength. Furthermore, the study population is primarily composed of children from Western countries. This article seeks to interpret the consensus to improve standardized use of melatonin for insomnia in Chinese children with ASD and NGD, and to provide a reference for the future development of localized evidence-based guidelines.

Objective:In order to reduce the trauma associated with fetal cardiopulmonary bypass(F-CPB), Our team plans to develop a minimally invasive F-CPB through a small incision in the right axilla. The efficacy of this technique will be verified by using a big experimental animal model, thereby laying the foundation for fetal cardiac surgery supported by F-CPB in the future.Methods:Ten pregnant sheep were divided into F-CPB group(n=5) and control group(n=5). After fasting for 24 h, fetal lambs in the F-CPB group underwent a right axillary incision to establish F-CPB running for 1 h; The control group of fetal lambs only expose heart 1 h without F-CPB. Collect blood sample for laboratory test at the CPB vehicle before(T0), 30 min(T1), and 1 h after F-CPB running(T2) for the F-CPB group and through Superior Vena Cava before(T0), 30 min(T1), and 1 h after F-CPB running(T2) for the control group.Results:The blood routine indicators such as RBC, HCT, and Hb in the F-CPB group of fetal lambs decreased significantly during F-CPB, and their distribution showed significant statistical differences compared with the control group( P<0.05). There were no significant statistical differences in blood gas indicators such as pH, PO 2, PCO 2, and lactate concentration between the F-CPB group and the control group( P>0.05). There was no statistically significant difference in the concentration of cTnI in fetal lamb serum at each time point( P>0.05). There were significant statistical differences( P<0.05) in the distribution of fetal lamb Alb, γ-GGT, CK concentration and cholesterol concentration at various time points in the F-CPB group compared with the control group in liver function examination. In addition, the distribution of BUN in fetal lambs showed a significant difference between the two groups( P=0.006). Conclusion:A minimally invasive F-CPB via small incision in the right axilla is safe and feasible. The experimental animal model has demonstrated that this technique has minimal impact on the vital organ functions and internal environment of fetal lambs, thereby laying the foundation for clinical fetal cardiac surgery in the future.

Melatonin is widely used as an over-the-counter medication to treat insomnia in children with autism spectrum disorder (ASD) and neurogenetic disorders (NGD). However, there is still a lack of research on its efficacy and safety, and clinical practice standards are to be established. In response, the International Pediatric Sleep Association (IPSA) convened an expert panel and developed a consensus statement:"Melatonin Use in Managing Insomnia in Children with Autism and Other Neurogenetic Disorders-an Assessment by the International Pediatric Sleep Association (IPSA)", which was published in Sleep Medicine, April 2024. The consensus focused on the efficacy and adverse effects of melatonin treatment for insomnia in children with ASD and NGD-including Smith-Magenis syndrome, Rett syndrome, Angelman syndrome, and tuberous sclerosis complex. It systematically reviews randomized controlled trials (RCTs) conducted between 2012 and 2022, and integrates current best clinical practices to formulate 10 consensus recommendations. Despite these contributions, the consensus has limitations: a small number of included RCTs, a lack of grading for evidence quality, and recommendation strength. Furthermore, the study population is primarily composed of children from Western countries. This article seeks to interpret the consensus to improve standardized use of melatonin for insomnia in Chinese children with ASD and NGD, and to provide a reference for the future development of localized evidence-based guidelines.

Objective The aim is to utilize CRISPR/Cas9 gene editing technology to construct Dmd gene mutant mice with a point mutation in exon 23 of the Dmd gene. Subsequently, the phenotypic changes of the mice in muscles and immune systems are analyzed and verified, providing an evaluation model for Duchenne muscular dystrophy and other related diseases.MethodsBased on the sequence characteristics of exon 23 of the Dmd gene, small guide RNA (sgRNA) was designed and synthesized. Cas9 mRNA, sgRNA fragments, and oligo donor DNA were microinjected into fertilized eggs of C57BL/6J mice. After transferring the fertilized eggs to surrogate mice, F0 generation mice were born. After mating with F0 generation mice, offspring mice were obtained, and Dmd gene positive mutant (Dmd^Mu/+) mice were obtained after genotype identification. Male hemizygous Dmd^Mu/+(Dmd^Mu/Y) mice were selected for phenotype validation. The body weight of live 3- and 9-month-old mice were recorded. Muscle tension was evaluated through the grid test. Hearts and semitendinosus muscles were collected, and the histopathological changes were observed using HE staining. Further, the expression of Dmd protein in muscle tissue of 9-month-old mice was analyzed by Western blotting.An acute inflammation model was established in Dmd^Mu/Y mice using lipopolysaccharide induction. Peripheral blood from the submandibular vein was collected, and the changes in the proportion of neutrophils and monocytes were detected by flow cytometry.Results The results of genome sequencing and Western blotting confirmed the successful construction of Dmd gene point mutant mice (Dmd^Mu/+ mice). Dmd protein expression was not detected in skeletal muscle and myocardium of Dmd^Mu/+ mice, and it was significantly reduced compared to wild-type C57BL/6J mice (P<0.05). Compared with wild-type mice of the same background, Dmd^Mu/Y mice at 3 and 9 months of age showed significant weight loss (P<0.01) and decreased muscle tension (P<0.05). 9-month-old Dmd^Mu/Y mice exhibited significant pathological changes in skeletal muscle and myocardium, including widening of intermuscular space. Under normal condition, compared with wild-type mice, the proportion of neutrophils and monocytes in the peripheral blood of 3-month-old Dmd^Mu/Y mice was significantly lower than that of wild-type mice (P<0.01). After lipopolysaccharide stimulation, the proportion of neutrophils in peripheral blood of 3-month-old Dmd^Mu/Y mice remained significantly lower compared to that of wild-type mice (P<0.01). The proportion of neutrophils in peripheral blood of 9-month-old Dmd^Mu/Y mice significantly decreased after lipopolysaccharide induction (P<0.01), with a trend of change observed in monocytes between groups.Conclusion The successful construction of the Dmd gene mutant mouse model has confirmed the vital function of Dmd gene in maintaining normal muscle tissue morphology and muscle tone. It preliminarily indicated that Dmd gene deletion could significantly reduce the proportion of neutrophils in peripheral blood, offering a new perspective for the study of immune system alterations in Duchenne muscular dystrophy patients.

The timely treatment of dental caries and pulp disease in primary teeth holds significant importance for maintaining children's oral health.Direct pulp capping(DPC)is a vital pulp treatment that involves covering the ex-posed pulp with bioactive materials to promote dentin bridge formation.DPC is commonly used in primary teeth with vi-tal pulp and mechanical pulp exposure not exceeding 1 mm.DPC offers advantages such as minimal invasiveness,com-fort,simplicity of operation and short chair-side time,making it suitable for pediatric dental clinical practice.Early stud-ies suggested negative treatment outcomes for DPC in primary teeth with carious pulp exposure.Over the years,there have been advancements in materials and technology demonstrating positive outcomes in the clinical research of prima-ry teeth with deep caries.However,due to the limited quality of related studies,DPC has not been widely recommended for the treatment of primary teeth with carious pulp exposure,and its widespread use needs further support by more high-quality evidence-based medical research.The success rate of DPC in primary teeth is influenced by factors including pulp status,clinical operations(such as isolation and caries removal),pulp capping material,cavity type,tooth position,coronal sealing,and dental fear.In clinical operation,dentists should accurately assess pulp status and minimize bacte-rial contamination.Mineral trioxide aggregate(MTA)is a DPC agent with relatively sufficient evidence and good thera-peutic effects,and the crown should be tightly sealed after pulp capping.Additionally,the effects of novel biocompati-ble materials such as iRoot BP Plus used in DPC of primary teeth,and the influence of other factors like hemostatic methods on the prognosis of affected teeth,need further exploration.

The timely treatment of dental caries and pulp disease in primary teeth holds significant importance for maintaining children's oral health.Direct pulp capping(DPC)is a vital pulp treatment that involves covering the ex-posed pulp with bioactive materials to promote dentin bridge formation.DPC is commonly used in primary teeth with vi-tal pulp and mechanical pulp exposure not exceeding 1 mm.DPC offers advantages such as minimal invasiveness,com-fort,simplicity of operation and short chair-side time,making it suitable for pediatric dental clinical practice.Early stud-ies suggested negative treatment outcomes for DPC in primary teeth with carious pulp exposure.Over the years,there have been advancements in materials and technology demonstrating positive outcomes in the clinical research of prima-ry teeth with deep caries.However,due to the limited quality of related studies,DPC has not been widely recommended for the treatment of primary teeth with carious pulp exposure,and its widespread use needs further support by more high-quality evidence-based medical research.The success rate of DPC in primary teeth is influenced by factors including pulp status,clinical operations(such as isolation and caries removal),pulp capping material,cavity type,tooth position,coronal sealing,and dental fear.In clinical operation,dentists should accurately assess pulp status and minimize bacte-rial contamination.Mineral trioxide aggregate(MTA)is a DPC agent with relatively sufficient evidence and good thera-peutic effects,and the crown should be tightly sealed after pulp capping.Additionally,the effects of novel biocompati-ble materials such as iRoot BP Plus used in DPC of primary teeth,and the influence of other factors like hemostatic methods on the prognosis of affected teeth,need further exploration.

The timely treatment of dental caries and pulp disease in primary teeth holds significant importance for maintaining children's oral health.Direct pulp capping(DPC)is a vital pulp treatment that involves covering the ex-posed pulp with bioactive materials to promote dentin bridge formation.DPC is commonly used in primary teeth with vi-tal pulp and mechanical pulp exposure not exceeding 1 mm.DPC offers advantages such as minimal invasiveness,com-fort,simplicity of operation and short chair-side time,making it suitable for pediatric dental clinical practice.Early stud-ies suggested negative treatment outcomes for DPC in primary teeth with carious pulp exposure.Over the years,there have been advancements in materials and technology demonstrating positive outcomes in the clinical research of prima-ry teeth with deep caries.However,due to the limited quality of related studies,DPC has not been widely recommended for the treatment of primary teeth with carious pulp exposure,and its widespread use needs further support by more high-quality evidence-based medical research.The success rate of DPC in primary teeth is influenced by factors including pulp status,clinical operations(such as isolation and caries removal),pulp capping material,cavity type,tooth position,coronal sealing,and dental fear.In clinical operation,dentists should accurately assess pulp status and minimize bacte-rial contamination.Mineral trioxide aggregate(MTA)is a DPC agent with relatively sufficient evidence and good thera-peutic effects,and the crown should be tightly sealed after pulp capping.Additionally,the effects of novel biocompati-ble materials such as iRoot BP Plus used in DPC of primary teeth,and the influence of other factors like hemostatic methods on the prognosis of affected teeth,need further exploration.