ObjectiveTo optimize the extraction method for polysaccharides from turnip（Brassica rapa）， and analyze and evaluate the primary structure of the isolated and purified turnip polysaccharide fraction（BP-1） and its hypoglycemic effects in diabetic zebrafish. MethodsTaking polysaccharide yield as the evaluation index， a semi-bionic extraction method was employed. Single-factor experiments and Box-Behnken response surface methodology were used to investigate three factors of solid-to-liquid ratio， extraction time and extraction temperature， in order to optimize the extraction process. BP-1 was isolated and purified using the Sevage method and DEAE-52 cellulose column chromatography. Structural characterization of the turnip polysaccharides was performed using ultraviolet-visible spectrophotometry（UV）， gas chromatography-mass spectrometry（GC-MS）， Congo red assay， and Fourier-transform infrared spectroscopy（FT-IR） to determine purity， monosaccharide composition， triple-helix structure， and functional groups. The microstructure of the polysaccharides was observed using scanning electron microscopy（SEM） and atomic force microscopy（AFM）. Zebrafish were divided into the blank group（adding E3 medium）， and BP-1-1， BP-1-10， BP-1-50， BP-1-200， BP-1-1 000 groups（adding BP-1 solutions at concentrations of 1， 10， 50， 200， 1 000 mg·L^-1， respectively）， and zebrafish embryos were subjected to a 96-hour exposure experiment. The maximum tolerated concentration of BP-1 in zebrafish was determined by evaluating its effects on phenotype， survival rate， malformation rate， and heart rate. Experimental animals were randomly divided into the blank group， model group， BP-1-10 group（10 mg·L^-1）， BP-1-50 group（50 mg·L^-1）， and BP-1-200 group（200 mg·L^-1）. The blank group was cultured in E3 medium， the model and treatment groups were induced to establish a diabetic model in 4 day-post-fertilization（dpf） zebrafish embryos using 10 g·L^-1 of glucose combined with 500 µmol·L^-1 of alloxan. The treatment groups received corresponding doses of BP-1 solution， while the blank and model groups received an equal volume of saline. Glucose and insulin（INS） levels were measured using enzyme-linked immunosorbent assay（ELISA） kits， the effects on the liver were observed by hematoxylin-eosin（HE） histopathological sections. The mRNA expression levels of glucagon（Glucagon）， insulin（Insa）， and phosphoenolpyruvate carboxykinase 1（PCK1） were detected with real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. ResultsThe optimized extraction conditions were determined as follows：solid-to-liquid ratio of 1∶40（g·mL^-1）， extraction time of 66 min， and extraction temperature of 79 ℃. Under these conditions， the yield of turnip polysaccharides was （10.34±0.96）%. UV analysis indicated that BP-1 contained no proteins or nucleic acids， GC-MS analysis revealed that BP-1 consisted of six monosaccharides（arabinose， rhamnose， ribose， mannose， galactose and glucose）. Congo red assay indicated that the molecular conformation did not exhibit a triple-helix structure， FT-IR analysis showed the presence of α-glycosidic bonds and uronic acids， SEM analysis revealed an irregular flaky structure with a flat and smooth surface， AFM analysis suggested that the aggregated structure might be formed by the entanglement of molecular chains and intramolecular hydrogen bonding. The maximum tolerated concentration of BP-1 in zebrafish over 96 h was determined to be 200 mg·L^-1. Pharmacodynamic results showed that， compared with the blank group， the model group exhibited significantly increased glucose levels and significantly decreased INS levels（P<0.01）. Compared with the model group， the BP-1-50 group significantly reduced glucose levels and increased INS levels（P<0.05）. Histopathological examination of liver tissue revealed that various doses of BP-1 had a certain reparative effect on damaged liver tissue. The liver tissue structure in the BP-1-200 group was nearly normal， with hepatocytes appearing plump. Real-time PCR results showed that， compared with the blank group， the model group exhibited significantly upregulated mRNA expressions of Glucagon and PCK1， and significantly downregulated mRNA expression of Insa（P<0.01）. Compared with the model group， the BP-1-50 and BP-1-200 groups showed significantly downregulated mRNA expressions of Glucagon and PCK1， and significantly upregulated mRNA expression of Insa（P<0.01）. ConclusionThe semi-bionic extraction method for turnip polysaccharides yields a high extraction rate， is simple to operate， has low costs， making it suitable for large-scale industrial production. BP-1 consists of six monosaccharides， contains α-glycosidic bonds and uronic acids， exhibits hypoglycemic activity， and provides a certain protective effect on the liver of alloxan-induced diabetic model zebrafish.

ObjectiveTo optimize the extraction method for polysaccharides from turnip（Brassica rapa）， and analyze and evaluate the primary structure of the isolated and purified turnip polysaccharide fraction（BP-1） and its hypoglycemic effects in diabetic zebrafish. MethodsTaking polysaccharide yield as the evaluation index， a semi-bionic extraction method was employed. Single-factor experiments and Box-Behnken response surface methodology were used to investigate three factors of solid-to-liquid ratio， extraction time and extraction temperature， in order to optimize the extraction process. BP-1 was isolated and purified using the Sevage method and DEAE-52 cellulose column chromatography. Structural characterization of the turnip polysaccharides was performed using ultraviolet-visible spectrophotometry（UV）， gas chromatography-mass spectrometry（GC-MS）， Congo red assay， and Fourier-transform infrared spectroscopy（FT-IR） to determine purity， monosaccharide composition， triple-helix structure， and functional groups. The microstructure of the polysaccharides was observed using scanning electron microscopy（SEM） and atomic force microscopy（AFM）. Zebrafish were divided into the blank group（adding E3 medium）， and BP-1-1， BP-1-10， BP-1-50， BP-1-200， BP-1-1 000 groups（adding BP-1 solutions at concentrations of 1， 10， 50， 200， 1 000 mg·L^-1， respectively）， and zebrafish embryos were subjected to a 96-hour exposure experiment. The maximum tolerated concentration of BP-1 in zebrafish was determined by evaluating its effects on phenotype， survival rate， malformation rate， and heart rate. Experimental animals were randomly divided into the blank group， model group， BP-1-10 group（10 mg·L^-1）， BP-1-50 group（50 mg·L^-1）， and BP-1-200 group（200 mg·L^-1）. The blank group was cultured in E3 medium， the model and treatment groups were induced to establish a diabetic model in 4 day-post-fertilization（dpf） zebrafish embryos using 10 g·L^-1 of glucose combined with 500 µmol·L^-1 of alloxan. The treatment groups received corresponding doses of BP-1 solution， while the blank and model groups received an equal volume of saline. Glucose and insulin（INS） levels were measured using enzyme-linked immunosorbent assay（ELISA） kits， the effects on the liver were observed by hematoxylin-eosin（HE） histopathological sections. The mRNA expression levels of glucagon（Glucagon）， insulin（Insa）， and phosphoenolpyruvate carboxykinase 1（PCK1） were detected with real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. ResultsThe optimized extraction conditions were determined as follows：solid-to-liquid ratio of 1∶40（g·mL^-1）， extraction time of 66 min， and extraction temperature of 79 ℃. Under these conditions， the yield of turnip polysaccharides was （10.34±0.96）%. UV analysis indicated that BP-1 contained no proteins or nucleic acids， GC-MS analysis revealed that BP-1 consisted of six monosaccharides（arabinose， rhamnose， ribose， mannose， galactose and glucose）. Congo red assay indicated that the molecular conformation did not exhibit a triple-helix structure， FT-IR analysis showed the presence of α-glycosidic bonds and uronic acids， SEM analysis revealed an irregular flaky structure with a flat and smooth surface， AFM analysis suggested that the aggregated structure might be formed by the entanglement of molecular chains and intramolecular hydrogen bonding. The maximum tolerated concentration of BP-1 in zebrafish over 96 h was determined to be 200 mg·L^-1. Pharmacodynamic results showed that， compared with the blank group， the model group exhibited significantly increased glucose levels and significantly decreased INS levels（P<0.01）. Compared with the model group， the BP-1-50 group significantly reduced glucose levels and increased INS levels（P<0.05）. Histopathological examination of liver tissue revealed that various doses of BP-1 had a certain reparative effect on damaged liver tissue. The liver tissue structure in the BP-1-200 group was nearly normal， with hepatocytes appearing plump. Real-time PCR results showed that， compared with the blank group， the model group exhibited significantly upregulated mRNA expressions of Glucagon and PCK1， and significantly downregulated mRNA expression of Insa（P<0.01）. Compared with the model group， the BP-1-50 and BP-1-200 groups showed significantly downregulated mRNA expressions of Glucagon and PCK1， and significantly upregulated mRNA expression of Insa（P<0.01）. ConclusionThe semi-bionic extraction method for turnip polysaccharides yields a high extraction rate， is simple to operate， has low costs， making it suitable for large-scale industrial production. BP-1 consists of six monosaccharides， contains α-glycosidic bonds and uronic acids， exhibits hypoglycemic activity， and provides a certain protective effect on the liver of alloxan-induced diabetic model zebrafish.

Objective To explore whether the environmental pollutant triclosan(TCS)has negative effects on the various biological characteristics of dental pulp stem cells(DPSCs),as well as the distribution and hazards of TCS in rat dental pulp tissue in vivo,which will provide a basis for the clinical application of DPSCs and the safety of TCS.Methods Tooth collection was approved by the Ethics Committee of Tianyou Hospital Affiliated to Wuhan University of Science and Technology.Human DPSCs were extracted,cultured,and identified.Up to 0.08 mmol/L of TCS was added to the in vitro culture medium of DPSCs.The proliferation ability of DPSCs was detected by CCK-8.The migration ability of DPSCs was detected via scratch assay.The differentiation ability of DPSCs was detected by inducing trilineage differenti-ation.The gene or protein expression levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-10(IL-10),inducible nitric oxide synthase(iNOS),and transforming growth factor-β(TGF-β)in DPSCs were detected.The level of reactive oxygen species(ROS)generated by DPSCs was analyzed using fluorescence staining.Changes in mitochondrial membrane potential of DPSCs were detected using a fluorescent probe.The activity of PI3K/Akt/mTOR,p38,and JNK pathways of DPSCs were detected.Animal experiments were approved by the Animal Ethics Committee of Wuhan University of Science and Technology.A rat model of short-term oral exposure to 50 mg/kg/d of TCS for 2 months was established,and the TCS concentration in the liver,brain,and dental pulp tissues of rats was detected through liquid chromatography-mass spectrometry.Results TCS at 0.02 mmol/L,0.04 mmol/L,and 0.08 mmol/L significantly inhibited the proliferation ability of human-derived DPSCs on the 5th and 7th days of contact.TCS at 0.04 mmol/L and 0.08 mmol/L significantly inhibited the migration ability and tri-lineage differentiation ability of DPSCs on the 3rd day of contact.TCS induced the gene or protein expression of proinflammatory factors including TNF-α,IL-1β,IL-6,and iNOS,induced the gene or protein expression of TGF-β,and inhibited the protein expression of anti-inflammatory factor IL-10.On day 1,TCS at 0.04 mmol/L and 0.08 mmol/L induced the production of ROS in DPSCs and reduced the mitochondrial membrane potential of DPSCs.On day 3,TCS at these levels inhibited PI3K/Akt/mTOR pathway activity and enhanced p38 pathway activity of DPSCs,without affecting the pathway activity of JNK.After short-term intragastric exposure of rats to TCS,TCS was detected in liver(430 ng/mL)and brain(41.4 ng/mL)tissues but not in the dental pulp.The TCS concentration was highest in the liver,but no obvious histopathological changes were observed.Conclusion TCS inhibits a variety of biological characteristics of DPSCs and poses a potential risk to the organism.No TCS exists in the dental pulp tissue of rats exposed to TCS for a brief period of time,and the health of the rats is not damaged.

Malocclusion,identified by the World Health Organization(WHO)as one of three major oral diseases,profoundly impacts the dental-maxillofacial functions,facial esthetics,and long-term development of～260 million children in China.Beyond its physical manifestations,malocclusion also significantly influences the psycho-social well-being of these children.Timely intervention in malocclusion can foster an environment conducive to dental-maxillofacial development and substantially decrease the incidence of malocclusion or reduce the severity and complexity of malocclusion in the permanent dentition,by mitigating the negative impact of abnormal environmental influences on the growth.Early orthodontic treatment encompasses accurate identification and treatment of dental and maxillofacial morphological and functional abnormalities during various stages of dental-maxillofacial development,ranging from fetal stages to the early permanent dentition phase.From an economic and societal standpoint,the urgency for effective early orthodontic treatments for malocclusions in childhood cannot be overstated,underlining its profound practical and social importance.This consensus paper discusses the characteristics and the detrimental effects of malocclusion in children,emphasizing critical need for early treatment.It elaborates on corresponding core principles and fundamental approaches in early orthodontics,proposing comprehensive guidance for preventive and interceptive orthodontic treatment,serving as a reference for clinicians engaged in early orthodontic treatment.

Tooth extraction is a common and widely employed therapeutic procedure in oral and maxillofacial surgery.Minimally invasive tooth extraction can reduce both physical and psychological trauma to the patients,and is widely recommended as a first-line clinical treatment.But currently no guidelines or consensus has been available to provide a systematic introduction of minimally invasive tooth extraction to guide the clinical practices.To address this issue,this consensus,based on a comprehensive literature review and clinical experiences of experts,systematically summarizes the indications,target patients,and contraindications of minimally invasive tooth extraction,the overall workflow of this procedure(preoperative preparation,surgical steps,postoperative management,postoperative instructions,medications,and follow-up),and its common postoperative complications to provide a comprehensive guidance for clinical application of this technique.

Tooth extraction is a common and widely employed therapeutic procedure in oral and maxillofacial surgery.Minimally invasive tooth extraction can reduce both physical and psychological trauma to the patients,and is widely recommended as a first-line clinical treatment.But currently no guidelines or consensus has been available to provide a systematic introduction of minimally invasive tooth extraction to guide the clinical practices.To address this issue,this consensus,based on a comprehensive literature review and clinical experiences of experts,systematically summarizes the indications,target patients,and contraindications of minimally invasive tooth extraction,the overall workflow of this procedure(preoperative preparation,surgical steps,postoperative management,postoperative instructions,medications,and follow-up),and its common postoperative complications to provide a comprehensive guidance for clinical application of this technique.

Objective:To determine whether the transepidermal water loss rate (TEWL) is correlated with the stratum corneum (SC) hydration level.Methods:Healthy children aged ≤ 17 years were enrolled from Medical Center for Public Health of Puning, 2 kindergartens and 2 primary schools, from October 2021 to June 2022. TEWL and SC hydration levels were measured on the left forearm and right anterior shank using a device for measuring skin physiological funcitons. Pearson correlation analysis was used to determine the correlations between TEWL and SC hydration levels in children of different ages and genders.Results:A total of 1 396 healthy children were enrolled, aged from 1 month to 17 years. Among them, 783 were male children and 613 were female children. In children aged 1 to < 12 months, no correlation was observed between TEWL and SC hydration levels on the forearms of male children, while TEWL was positively correlated with SC hydration levels on the anterior shanks of male children, as well as on the forearm and anterior shanks of female children ( r = 0.283, 0.404, 0.420, respectively, all P < 0.05) . In children aged 1 to 2 years, positive correlations were observed between the above two indicators on the anterior shanks of male children and forearms of female children ( r = 0.370, 0.419, respectively, both P < 0.01) , while there were no correlations between the two indicators on the anterior shanks of female children or forearms of male children. Positive correlations were observed between TEWL and SC hydration levels on both the forearms and anterior shanks of female children and the forearms of male children aged 3 to 5 years and 6 to 11 years ( r values ranging from 0.172 to 0.293, all P < 0.05) , but not on the anterior shanks of male children aged from 6 to 11 years. The group aged 12 to 17 years exhibited significantly positive correlations between TEWL and SC hydration levels on both the anterior shanks and forearms of male and female children ( r values ranging from 0.269 to 0.485, all P < 0.001) . Conclusion:SC hydration levels are positively correlated with TEWL on the anterior shanks and forearms of healthy children, and the degree of correlation tends to increase with age.

Cell therapy based on mesenchymal stem cells (MSCs) has been a hot research topic in recent years, including the traditional cell therapy strategy based on living cells and the new cell-free therapy strategy based on soluble proteins or bioactive molecules such as extracellular vesicles (EVs). At present, MSC-induced cells have mature functions and specific structures, and insitu transplantation combined with biomaterials or organic technology has greatly improved the settlement rate and function. On the other hand, as the large-scale culture technique and EVs separation technique evolve, it is possible to obtain a large number of pure EVs, and EVs are gradually becoming a hot spot of current research. An increasing number of studies have shown that the therapeutic effect of MSCs not only occurs by implantation and differentiation but also manifests as the paracrine effect of MSCs. In this review, we discuss the emerging outcomes of cell therapies and acellular therapies to alleviate these pathological conditions.

Objective:To analyze the diagnostic value of cell-free plasma metagenomic next-generation sequencing (mNGS) pathogen identification for severe aplastic anemia (SAA) bloodstream infection.Methods:From February 2021 to February 2022, mNGS and conventional detection methods (blood culture, etc.) were used to detect 33 samples from 29 consecutive AA patients admitted to the Anemia Diagnosis and Treatment Center of the Hematology Hospital of the Chinese Academy of Medical Sciences to assess the diagnostic consistency of mNGS and conventional detection, as well as the impact on clinical treatment benefits and clinical accuracy.Results:①Among the 33 samples evaluated by mNGS and conventional detection methods, 25 cases (75.76%) carried potential pathogenic microorganisms. A total of 72 pathogenic microorganisms were identified from all cases, of which 65 (90.28%) were detected only by mNGS. ②All 33 cases were evaluated for diagnostic consistency, of which 2 cases (6.06%) were Composite, 18 cases (54.55%) were mNGS only, 2 cases (6.06%) were Conventional method only, 1 case (3.03%) was both common compliances (mNGS/Conventional testing) , and 10 cases (30.3%) were completely non-conforming (None) . ③All 33 cases were evaluated for clinical treatment benefit. Among them, 8 cases (24.24%) received Initiation of targeted treatment, 1 case (3.03%) received Treatment de-escalation, 13 cases (39.39%) received Confirmation, and the remaining 11 cases (33.33%) received No clinical benefit. ④ The sensitivity of 80.77%, specificity of 70.00%, positive predictive value of 63.64%, negative predictive value of 84.85%, positive likelihood ratio of 2.692, and negative likelihood ratio of 0.275 distinguished mNGS from conventional detection methods (21/12 vs 5/28, P<0.001) . Conclusion:mNGS can not only contribute to accurately diagnosing bloodstream infection in patients with aplastic anemia, but can also help to guide accurate anti-infection treatment, and the clinical accuracy is high.

Exosomes are phospholipid bilayer vesicles secreted by living cells that can carry a variety of signaling molecules, such as RNA, DNA, protein, and lipids. Exosomes play a role in the transmission of signaling molecules between cells, thus regulating many physiological and pathological processes. The methods of extracting exosomes include differential centrifugation, density gradient centrifugation, exclusion chromatography, ultrafiltration, coprecipitation, polymer immune affinity, microfluidic separation technology, etc. Each of these extraction technologies has advantages and disadvantages; however, there is no unified international standard. In addition, the expression of specific proteins and genetic material of exosomes from different cell sources are different; thus, their expression characteristics and functions are also distinctive. Based on this situation, research on exosomes is limited to preclinical studies, and difficulties and challenges still exist in clinical application. This paper summarizes the progress of research in the field of exosomes, to understand the characteristics, modification and application of exosomes from different cell sources, and to summarize their advantages and disadvantages as well as challenges, which can help researchers better understand and master the performance of exosomes. Furthermore, improvement of standard procedures in the extraction and manufacturing of exosomes is important, as it will provide a reference for researchers to carry out exosome-related translational clinical research.