ObjectiveThis study aims to explore the mechanism by which Yishen Huoxue Tongqiao Formula improves metabolism-neuroplasticity and treats unilateral vestibular labyrinth destruction by regulating the metabolic balance of glutamate （Glu）/γ-aminobutyric acid （GABA）. Methods48 Sprague-Dawley （SD） adult rats were randomly divided into the sham operation group， model group， Yishen Huoxue Tongqiao Formula groups with low， medium， and high doses （9.20， 18.39， 36.78 g·kg^-1）， and betahistine group （1.62 mg·kg^-1）. A unilateral vestibular labyrinth destruction （vestibular dysfunction） model was established by intratympanic injection of chloroform into the right ear， while the control group received intratympanic injection of normal saline. Drugs were administered once daily for seven consecutive days. During the period， behavioral tests were performed to evaluate the behaviors of rats after unilateral vestibular labyrinth destruction. Hematoxylin-eosin （HE） staining and Nissl staining were used to observe the neuronal morphology in the medial vestibular nucleus. Golgi staining was employed to assess the number of dendritic spines of neurons in the medial vestibular nucleus. Ultra-performance liquid chromatography-tandem mass spectrometry （LC-ESI-MS/MS） was utilized to detect Glu/GABA. Immunofluorescence and immunohistochemistry were used to detect the expressions of neuronal nuclei （NeuN）， growth-associated protein 43 （GAP-43）， and glial fibrillary acidic protein （GFAP）. Western blot and real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） were applied to determine the expressions of glutamate-immunoreactive （Glu-IR）， GABA， GFAP， postsynaptic density protein 95 （PSD-95）， and GAP-43. ResultsCompared with the sham operation group， the model group presented with head deviation， balance disorder， increased tail suspension score， nuclear consolidation of medial vestibular nerve neurons， and decreased Nissl bodies （P<0.01）. The number of dendritic spines in neurons and NeuN-positive cells decreased. The content of Glu decreased. The content of GABA increased （Glu/GABA decreased）. The expression of GAP-43 was down-regulated， and GFAP was up-regulated （P<0.05， P<0.01）. The expressions of Glu-IR， PSD-95， and GAP-43 proteins， as well as Glu-IR mRNA decreased， while the expressions of GABA and GFAP proteins and mRNA increased （P<0.05， P<0.01）. Compared with those in the model group， the head deviation， imbalanced behavior， and tail suspension scores in each treatment group decreased， with alleviated neuronal injury and recovered Nissl bodies （P<0.01）. The number of dendritic spines of neurons increased， and the number of NeuN-positive cells rebounded. The content of Glu increased， and the content of GABA decreased （Glu/GABA increased）. GFAP was down-regulated， and GAP-43 was up-regulated （P<0.05， P<0.01）. The expressions of Glu-IR， PMD-95， and GAP-43 proteins， as well as Glu-IR mRNA increased， while the expressions of GABA and GFAP proteins and mRNA decreased. The effect was more significant in the high-dose group （P<0.01）. ConclusionThe Yishen Huoxue Tongqiao Formula can alleviate vestibular dysfunction， and its mechanism may be associated with regulating the metabolic balance of Glu/GABA， mitigating neural damage， improving synaptic plasticity （promoting GAP-43 expression and inhibiting GFAP expression）， and facilitating vestibular compensation.

To construct a subgenomic replication subsystem of foot-and-mouth disease virus(FM-DV)carrying fluorescent protein(UnaG),the full-length cDNA plasmid of FMDV was employed as the template.A few of structural and non-structural protein genes of FMDV were removed by double-enzyme digestion.By substituting reporter gene sequences expressing the green fluorescent protein UnaG for specific structural protein sequences of FMDV,FMDV-UnaG replicators were successfully created.After PCR and sequencing,linearized FMDV-UnaG replicons were transfected into BSR/T7 cells expressing T7 RNA polymerase,and the fluorescence signal was observed through fluorescence microscopy and laser confocal technique.The results demonstrated that the constructed FMDV-UnaG replicons could effectively express UnaG protein,and the protein colo-calized with FMDV 3A protein.Additionally,Western blot and RT-qPCR also detected that the replicator RNA could express the non-structural proteins of the virus and replicate autonomously in BSR/T7 cells,respectively.In conclusion,the successful construction of FMDV-UnaG sub-genomic replicators offers a favorable tool for further research on the replication and translation mechanism of FMDV and the development of vaccine vectors.

Clear aligner treatment is a novel technique in current orthodontic practice. Distinct from traditional fixed orthodontic appliances, clear aligners have different material features and biomechanical characteristics and treatment efficiencies, presenting new clinical challenges. Therefore, a comprehensive and systematic description of the key clinical aspects of clear aligner treatment is essential to enhance treatment efficacy and facilitate the advancement and wide adoption of this new technique. This expert consensus discusses case selection and grading of treatment difficulty, principle of clear aligner therapy, clinical procedures and potential complications, which are crucial to the clinical success of clear aligner treatment.
Humans ; Consensus ; Orthodontic Appliance Design ; Orthodontic Appliances, Removable ; Tooth Movement Techniques/methods* ; Malocclusion/therapy* ; Orthodontics, Corrective/instrumentation*

To construct a subgenomic replication subsystem of foot-and-mouth disease virus(FM-DV)carrying fluorescent protein(UnaG),the full-length cDNA plasmid of FMDV was employed as the template.A few of structural and non-structural protein genes of FMDV were removed by double-enzyme digestion.By substituting reporter gene sequences expressing the green fluorescent protein UnaG for specific structural protein sequences of FMDV,FMDV-UnaG replicators were successfully created.After PCR and sequencing,linearized FMDV-UnaG replicons were transfected into BSR/T7 cells expressing T7 RNA polymerase,and the fluorescence signal was observed through fluorescence microscopy and laser confocal technique.The results demonstrated that the constructed FMDV-UnaG replicons could effectively express UnaG protein,and the protein colo-calized with FMDV 3A protein.Additionally,Western blot and RT-qPCR also detected that the replicator RNA could express the non-structural proteins of the virus and replicate autonomously in BSR/T7 cells,respectively.In conclusion,the successful construction of FMDV-UnaG sub-genomic replicators offers a favorable tool for further research on the replication and translation mechanism of FMDV and the development of vaccine vectors.

Objective To understand the current situation of breastfeeding duration in children aged 0-5 years in Yunnan Province,and to explore the influencing factors of breastfeeding duration.Methods Using the data of the 6th National Health Service Survey in Yunnan Province,1582 children aged 0～5 years in Yunnan Province were selected as the research subjects,and the Kaplan-Meier method and Cox regression were used to analyze the influencing factors of breastfeeding duration.Results The mean duration of breastfeeding for children aged 0～5 years in Yunnan Province was 9.29 months,and region,time of complementary food addition,time of suckling and family income were the main factors influencing the duration of breastfeeding.Conclusion The duration of breastfeeding for children aged 0～5 years in Yunnan Province deviates significantly from the recommendations provided by both the World Health Organization(WHO)and China's child breastfeeding guidelines.Given the current situation,the relevant departments must enhance their focus on this issue.

In the field of dental aesthetics,digital aesthetic design plays a crucial role in helping dentists to predict treatment outcomes vis-ually,as well as in enhancing the consistency of knowledge and understanding of aesthetic goals between dentists and patients.It serves as the foundation for achieving ideal aesthetic effects.However,there is no clear standard for this digital process currently in China and abroad.Many dentists lack of systematic understanding of how to carry out digital aesthetic design for treatment.To establish standardized processes for dental aesthetic design and to improve the homogeneity of treatment outcomes,Chinese Society of Digital Dental Industry(CSD-DI)convened domestic experts in related field to compile this consensus.This article elaborates on the key aspects of digital aesthetic data collection,integration steps,and the digital aesthetic design process.It also formulates a decision tree for dental aesthetics at macro level and outlines corresponding workflows for various clinical scenarios,serving as a reference for clinicians.

Atherosclerosis is a chronic vascular inflammatory disease caused by abnormal lipid metabolism.The formation of lipid-rich foam cells acts as the initial trigger for development of atherosclerotic lesions.Recent studies have shown that lipophagy,a form of selective autophagy,can selectively degrade lipid droplets stored intracellularly and promote cholesterol efflux through the autophagic lysosomal pathway.As a result,intracellular lipid accumulation is re-duced and foaming is inhibited,making lipophagy a potential new target for current anti-atherosclerosis therapy.This arti-cle reviews the crucial role and molecular mechanism of lipophagy in the link between lipid metabolism and atherosclero-sis.Its objective is to outline the regulatory mechanism of lipophagy and present fresh insights for the treatment of athero-sclerotic diseases.

OBJECTIVE To establish a method for concentration determination of caffeine and its three metabolites， theophylline， paraxanthine and theobromine in urine， and apply it in clinical practice. METHODS Using caffeine-13C3-d3 as internal standard （IS）， and the urine samples were protein precipitated with acetonitrile； HPLC-MS/MS method was adopted to determine the concentrations of caffeine and its three metabolites. The determination was performed on Waters ACQUITY UPLC® BEH HILIC column with mobile phase consisting of 60 mmol/L ammonium acetate （A）-acetonitrile （B） （gradient elution） at the flow rate of 0.5 mL/min. The column temperature was set at 38 ℃ ， and the sample size was 2 μL. The electrospray ionization detection was operated in a positive mode by multiple reaction monitoring. The detection ions for quantitative analysis were m/z 195.1→110.0 for caffeine， m/z 181.1→124.0 for theophylline， m/z 181.1→124.0 for paraxanthine， m/z 181.1→138.0 for theobromine， and m/z 198.1→ 140.1 for IS. The above method was used to determine the concentrations of caffeine and its three metabolites in the urine of 19 infants with apnea of prematurity （AOP）. RESULTS The linear ranges of mass concentration of caffeine， theophylline， paraxanthin and theobromine were 0.200-200， 0.050-50.0，0.050 0-50.0， and 0.100-100 μg/mL， respectively. The lower limits of quantification were 0.200， 0.050， 0.050 and 0.100 μg/mL （r＞0.990）， respectively. RSDs of intra-day and intra- day precision were not above 10.37%， and matrix factors were 85.68%-109.90%； extraction recoveries were 93.53%-109.40% （RSD≤15%）， and RSDs of stability tests were all lower than 15%. The concentrations of caffeine and its three metabolites in the urine of 19 cases were （27.346±7.951）， （0.351±0.223）， （0.428±0.395） and （0.472±0.374） μg/mL， respectively. CONCLUSIONS The established HPLC-MS/MS method is simple， sensitive and can be used for the determination of caffeine and its three metabolites in urine samples of AOP.

This paper summarized professor ZHANG Boli's experience in treating stubborn bi （痹） with the herbal pair of Ruxiang （Olibanum）- Moyao （Myrrha）. The basic pathogenesis of stubborn bi is channel and collateral stasis and obstruction. Ruxiang and Moyao are thus used in mutual reinforcement to rectify qi and diffuse bi， activate blood and relieve pain， thereby removing static and obstructed qi and blood， unblocking the obstructed channels and colla-terals， which is especially suitable for stubborn bi caused by channel and collateral obstruction. In clinical practice， the herbal pair of Ruxiang-Moyao is used together with qi-moving and blood-activating medicinals to treat chest bi by expelling stasis and diffusing stagnation， dissipating cold and unblocking vessels. To treat long-term wither and weakness in late stage of stroke， the medicinals of boosting qi and invigorating blood， unblocking channels and venting collaterals can be added to the herbal pair so as to soothe and drain vessels and collaterals， harmonize and regulate qi and blood. Simiao Yongan Decoction （四妙勇安汤） can be integrated in the treatment of vessel bi by moving qi and dissolving stasis， and for the long-term stubborn vessel bi， integrated internal and external treatment is suggested by external use of Ruxiang-Moyao to vent bi with aromatics. Moreover， it is emphasized to use the herbal pair of Ruxiang-Moyao in accordance with indications and cautions.

OBJECTIVE To prepare apigenin silk fibroin（API@SF）nanoparticles and to evaluate their safety and anti-tumor activity. METHODS API@SF nanoparticles were prepared by nanoprecipitation method ，and their morphology ，particle size ，Zeta potential，drug loading amount and in vitro release were characterized. The safety of nanoparticles was evaluated by hemolysis test and HE staining. MTT assay was adopted to evaluate inhibitory effects of API@SF nanoparticles on breast cancer 4T1 cells in mice. RESULTS The prepared API@SF nanoparticles were spherical with uniform distribution. The average particle size was 406.61 nm， the polydispersity index was 0.154，the Zeta potential was －18.4 mV，and the average drug-loading amount was 5.20％. The in vitro release results showed that the release rate of the nanoparticles was relatively fast in the release medium of pH 5.0 and relatively slow in the release medium of pH 7.4. Results of hemolysis test and HE staining showed that the nanoparticles had good biocompatibility. Results of MTT assay showed that the inhibitory effect of API@SF nanoparticles on 4T1 cells was significantly higher than that of API raw materials （P＜0.05），and its mechanism may be related to increasing the level of reactive oxygen species in cells. CONCLUSIONS API@SF nanoparticles are prepared successfully ，which possess good safety and anti-tumor activity.