ObjectiveTo explore the mechanism by which Yizhi Qingxin prescription improves mitochondrial dysfunction in Alzheimer's disease （AD） through regulating mitochondrial Ca²⁺ homeostasis and kinetic balance based on the protein kinase A （PKA）/calcineurin （CaN） signaling pathway. MethodsSixty three-month-old amyloid precursor protein （APP）/presenilin 1 （PS1） double transgenic mice were randomly divided into a model group， a donepezil group（0.65 mg·kg^-1）， a low-dose Yizhi Qingxin prescription group （YQF-L，2.6 g·kg^-1）， a medium-dose Yizhi Qingxin prescription group （YQF-M，5.2 g·kg^-1）， and a high-dose Yizhi Qingxin prescription group （YQF-H，10.4 g·kg^-1）， with 12 mice in each group. Twelve C57BL/6J mice with the same genetic background served as a normal group. Each treatment group received gavage administration daily， with the model and normal groups receiving equal volume of physiological saline. Intervention continued for 12 consecutive weeks. The learning and memory abilities of the mice were assessed using the novel object recognition （NOR） and Morris water maze （MWM） tests. Hematoxylin-eosin （HE）/Nissl staining was used to observe histopathological changes in the hippocampus. Transmission electron microscopy （TEM） was used to observe mitochondrial ultrastructure. Fluo-4 acetoxymethyl ester （Fluo-4 AM） Ca²⁺ probe was used to measure intracellular Ca²⁺ concentration in brain tissue. Western blot was used to determine the protein expression of PKA， CaN， sodium/calcium/lithium exchanger （NCLX）， mitochondrial calcium uniporter （MCU）， calmodulin （CaM）， dynamin-related protein 1 （Drp1）， and phosphorylated dynamin-related protein 1 （serine 637 site） ［p-Drp1（S637）］ in the hippocampus. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to measure the expression of PKA， CaN， CaM， NCLX， MCU， and Drp1 mRNAs. ResultsCompared with those in the normal group， the recognition index （RI） of the model group decreased （P0.01）， and the number of crossings through the original platform area， the duration of stay in the target quadrant， and the distance were reduced （P0.01）. The protein expression of PKA， NCLX， and p-DRP1 （ser637） significantly decreased （P0.05）， and the mRNA expression of PKA and NCLX significantly decreased （P0.05）. The escape latency （EL） was prolonged （P0.05）， and the intracellular Ca²⁺ level significantly increased （P0.01）. The protein expression of CaN， CaM， MCU， and Drp1， as well as the mRNA expression of CaN， MCU， and Drp1， significantly increased （P0.05）. After intervention with Donepezil and Yizhi Qingxin prescription， compared with that in the model group， the RI of the treatment group significantly increased （P0.05）， and the number of crossings through the platform and the duration of stay in the target quadrant significantly increased （P0.05）. The protein expression of PKA， NCLX， and p-Drp1 （ser637） and the mRNA expression of PKA and NCLX significantly increased （P0.05）. On the 4th and 5th days， the EL was shortened （P0.05）， and the intracellular Ca²⁺ level decreased （P0.05）. The protein expression of CaN， CaM， MCU， and Drp1 and the mRNA expression of CaN， MCU， and Drp1 significantly decreased （P0.05）. ConclusionYizhi Qingxin prescription regulates the PKA/CaN pathway， upregulates the expression of PKA， NCLX， and p-Drp1 （ser637） proteins， reduces the expression of CaN， CaM， MCU， and Drp1 proteins， and regulates Ca²⁺ homeostasis and mitochondrial dynamic balance， thereby enhancing the spatial learning and memory abilities of AD mice.

ObjectiveTo explore the mechanism by which Yizhi Qingxin prescription improves mitochondrial dysfunction in Alzheimer's disease （AD） through regulating mitochondrial Ca²⁺ homeostasis and kinetic balance based on the protein kinase A （PKA）/calcineurin （CaN） signaling pathway. MethodsSixty three-month-old amyloid precursor protein （APP）/presenilin 1 （PS1） double transgenic mice were randomly divided into a model group， a donepezil group（0.65 mg·kg^-1）， a low-dose Yizhi Qingxin prescription group （YQF-L，2.6 g·kg^-1）， a medium-dose Yizhi Qingxin prescription group （YQF-M，5.2 g·kg^-1）， and a high-dose Yizhi Qingxin prescription group （YQF-H，10.4 g·kg^-1）， with 12 mice in each group. Twelve C57BL/6J mice with the same genetic background served as a normal group. Each treatment group received gavage administration daily， with the model and normal groups receiving equal volume of physiological saline. Intervention continued for 12 consecutive weeks. The learning and memory abilities of the mice were assessed using the novel object recognition （NOR） and Morris water maze （MWM） tests. Hematoxylin-eosin （HE）/Nissl staining was used to observe histopathological changes in the hippocampus. Transmission electron microscopy （TEM） was used to observe mitochondrial ultrastructure. Fluo-4 acetoxymethyl ester （Fluo-4 AM） Ca²⁺ probe was used to measure intracellular Ca²⁺ concentration in brain tissue. Western blot was used to determine the protein expression of PKA， CaN， sodium/calcium/lithium exchanger （NCLX）， mitochondrial calcium uniporter （MCU）， calmodulin （CaM）， dynamin-related protein 1 （Drp1）， and phosphorylated dynamin-related protein 1 （serine 637 site） ［p-Drp1（S637）］ in the hippocampus. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to measure the expression of PKA， CaN， CaM， NCLX， MCU， and Drp1 mRNAs. ResultsCompared with those in the normal group， the recognition index （RI） of the model group decreased （P0.01）， and the number of crossings through the original platform area， the duration of stay in the target quadrant， and the distance were reduced （P0.01）. The protein expression of PKA， NCLX， and p-DRP1 （ser637） significantly decreased （P0.05）， and the mRNA expression of PKA and NCLX significantly decreased （P0.05）. The escape latency （EL） was prolonged （P0.05）， and the intracellular Ca²⁺ level significantly increased （P0.01）. The protein expression of CaN， CaM， MCU， and Drp1， as well as the mRNA expression of CaN， MCU， and Drp1， significantly increased （P0.05）. After intervention with Donepezil and Yizhi Qingxin prescription， compared with that in the model group， the RI of the treatment group significantly increased （P0.05）， and the number of crossings through the platform and the duration of stay in the target quadrant significantly increased （P0.05）. The protein expression of PKA， NCLX， and p-Drp1 （ser637） and the mRNA expression of PKA and NCLX significantly increased （P0.05）. On the 4th and 5th days， the EL was shortened （P0.05）， and the intracellular Ca²⁺ level decreased （P0.05）. The protein expression of CaN， CaM， MCU， and Drp1 and the mRNA expression of CaN， MCU， and Drp1 significantly decreased （P0.05）. ConclusionYizhi Qingxin prescription regulates the PKA/CaN pathway， upregulates the expression of PKA， NCLX， and p-Drp1 （ser637） proteins， reduces the expression of CaN， CaM， MCU， and Drp1 proteins， and regulates Ca²⁺ homeostasis and mitochondrial dynamic balance， thereby enhancing the spatial learning and memory abilities of AD mice.

ObjectiveTo investigate the neuroprotective mechanism of baicalein （BAI） on Parkinson's disease （PD） model rats by regulating endoplasmic reticulum stress pathway. MethodsSeventy-two Sprague-Dawley （SD） rats were randomly divided into normal group， model group， BAI low-dose group （80 mg·kg^-1）， medium-dose group （120 mg·kg^-1）， high-dose group （160 mg·kg^-1）， and levodopa-benserazide group （51 mg·kg^-1）， with 12 rats per group. Except for the normal group， PD rat models were established by subcutaneous injection of rotenone solution （2 mg·kg^-1） into the neck back of rats in the rest of groups for consecutive 28 days. Concurrently， rats in all groups received corresponding drugs via gavage for 28 days. After treatment， behavioral changes were assessed by using the open field and pole climbing tests. Neuronal pathology and apoptosis in the substantia nigra were observed via hematoxylin-eosin （HE） staining and TdT-mediated dUTP nick-end labeling （TUNEL） assay. α-Synuclein and tyrosine hydroxylase （TH） expressions were detected by immunohistochemistry （IHC）. Inflammatory factors such as interleukin-6 （IL-6）， interleukin-1β （IL-1β）， and tumor necrosis factor-α （TNF-α） were measured by enzyme-linked immunosorbent assay （ELISA）. RNA-like endoplasmic reticulum kinase （PERK）， activating transcription factor 4 （ATF4）， C/EBP-homologous protein （CHOP）， and Bcl-2-associated X protein （Bax） expressions were analyzed by Western blot. ResultsCompared with the normal group， the model group exhibited significantly reduced locomotion distance （P<0.01） and elevated pole-climbing scores （P<0.01）， with increased neuronal apoptosis rate （P<0.01）， significantly enhanced α-Synuclein expression （P<0.01）， decreased TH expression （P<0.01）， upregulated release of inflammatory factors （P<0.05，P<0.01）， and increased protein expressions of PERK/ATF4 pathway proteins and pro-apoptotic Bax （P<0.05，P<0.01）. Compared with the model group， medium/high-dose BAI groups and levodopa-benserazide group showed obviously improved motor function （P<0.05，P<0.01）， reduced pole-climbing scores （P<0.05）， decreased neuronal apoptosis （P<0.01）， downregulated α-Synuclein expression （P<0.01）， upregulated TH expression （P<0.05，P<0.01）， suppressed release of inflammatory factors （P<0.05，P<0.01）， and decreased protein expressions of PERK/ATF4 pathway proteins and pro-apoptotic Bax （P<0.05，P<0.01）. ConclusionBAI reduces the release of neuroinflammatory factors and neuronal apoptosis to improve the neurological function of PD model rats， and its mechanism may be related to alleviating endoplasmic reticulum stress and apoptosis by regulating the PERK/ATF4 pathway.

ObjectiveTo investigate the neuroprotective mechanism of baicalein （BAI） on Parkinson's disease （PD） model rats by regulating endoplasmic reticulum stress pathway. MethodsSeventy-two Sprague-Dawley （SD） rats were randomly divided into normal group， model group， BAI low-dose group （80 mg·kg^-1）， medium-dose group （120 mg·kg^-1）， high-dose group （160 mg·kg^-1）， and levodopa-benserazide group （51 mg·kg^-1）， with 12 rats per group. Except for the normal group， PD rat models were established by subcutaneous injection of rotenone solution （2 mg·kg^-1） into the neck back of rats in the rest of groups for consecutive 28 days. Concurrently， rats in all groups received corresponding drugs via gavage for 28 days. After treatment， behavioral changes were assessed by using the open field and pole climbing tests. Neuronal pathology and apoptosis in the substantia nigra were observed via hematoxylin-eosin （HE） staining and TdT-mediated dUTP nick-end labeling （TUNEL） assay. α-Synuclein and tyrosine hydroxylase （TH） expressions were detected by immunohistochemistry （IHC）. Inflammatory factors such as interleukin-6 （IL-6）， interleukin-1β （IL-1β）， and tumor necrosis factor-α （TNF-α） were measured by enzyme-linked immunosorbent assay （ELISA）. RNA-like endoplasmic reticulum kinase （PERK）， activating transcription factor 4 （ATF4）， C/EBP-homologous protein （CHOP）， and Bcl-2-associated X protein （Bax） expressions were analyzed by Western blot. ResultsCompared with the normal group， the model group exhibited significantly reduced locomotion distance （P<0.01） and elevated pole-climbing scores （P<0.01）， with increased neuronal apoptosis rate （P<0.01）， significantly enhanced α-Synuclein expression （P<0.01）， decreased TH expression （P<0.01）， upregulated release of inflammatory factors （P<0.05，P<0.01）， and increased protein expressions of PERK/ATF4 pathway proteins and pro-apoptotic Bax （P<0.05，P<0.01）. Compared with the model group， medium/high-dose BAI groups and levodopa-benserazide group showed obviously improved motor function （P<0.05，P<0.01）， reduced pole-climbing scores （P<0.05）， decreased neuronal apoptosis （P<0.01）， downregulated α-Synuclein expression （P<0.01）， upregulated TH expression （P<0.05，P<0.01）， suppressed release of inflammatory factors （P<0.05，P<0.01）， and decreased protein expressions of PERK/ATF4 pathway proteins and pro-apoptotic Bax （P<0.05，P<0.01）. ConclusionBAI reduces the release of neuroinflammatory factors and neuronal apoptosis to improve the neurological function of PD model rats， and its mechanism may be related to alleviating endoplasmic reticulum stress and apoptosis by regulating the PERK/ATF4 pathway.

Abstract Myopia is a major global public health concern, characterized by high prevalence among children and adolescents, frequent ocular complications, and multifactorial etiology. One potential contributing factor is serum vitamin D level. The paper summarizes epidemiological studies examining the relationship between serum vitamin D levels and myopia among children and adolescents. It further discusses possible biological mechanisms and future research prospects in order to elucidate the relationship between vitamin D levels and myopia in children and adolescents, and to provide a theoretical basis for effective prevention/control interventions.

OBJECTIVE To observe the effects of Modified shaoyao gancao decoction on intestinal transit function， intestinal flora and the contents of metabolites [γ aminobutyric acid （GABA） and 5-hydroxytryptamine （5-HT）] in slow transit constipation （STC） rats. METHODS SD rats were randomly divided into blank group （10 rats） and modeling group （30 rats）， with half male and half female. The STC model was established by intragastric administration of Compound diphenoxylate tablets in the modeling group. The successfully modeled rats were randomly divided into model group， Modified shaoyao gancao decoction group [56 g/（kg·d）， calculated by crude drug] and positive control group [lactulose 2.09 g/（kg·d）]， with 10 rats in each group. Each administration group was given relevant medicine intragastrically， the blank group and model group received an equivalent volume of normal saline， once a day， for 14 consecutive days. During the experiment， the general situation of rats was observed in each group. After the last medication， the body weight was measured， and the Bristol score was used to evaluate the fecal characteristics. The fecal moisture content， intestinal propulsion rate， and the contents of GABA and 5-HT in intestinal content were detected； the diversity of intestinal flora in intestinal contents was investigated， and the correlation between the contents of GABA， 5-HT and relative abundance of microbiota was analyzed. RESULTS Compared with the model group， general conditions such as small body shape， sparse and rough fur， and slow movement were all improved in Modified shaoyao gancao decoction body weight， Bristol score， fecal moisture content，intestinal propulsion rate， 5-HT content， Chao1 index and Shannon index were increased significantly， while GABA content and Simpson index were decreased significantly （P＜0.05）. The intestinal flora of rats in the Modified shaoyao gancao decoction group could be classified as the same as the blank group， but was far from the model group； the relative abundances of Desulfobacterota， Firmicutes and Bacteroidota in this group showed a tendency of pull back， but the differences were not statistically significant compared to model group （P＞0.05）. Desulfobacterota was an intergroup differential factor （P＜0.05）. The content of GABA was negatively correlated with the relative abundance of Bacteroidota， Cyanobacteria， Patescibacteria and Actinobacteriota （P＜0.05）. The content of 5-HT was positively correlated with the relative abundance of Campilobacterota （P＜0.05）. CONCLUSIONS Modified shaoyao gancao decoction can improve the fecal properties and intestinal motility of STC rats. Its mechanism may be related to improving intestinal flora and then affecting the contents of GABA and 5-HT in intestinal contents. In addition， the contents of GABA and 5-HT may be significantly correlated with the relative abundance of specific bacterial phyla such as Bacteroidota and Campilobacterota.

OBJECTIVE To observe the effects of Modified shaoyao gancao decoction on intestinal transit function， intestinal flora and the contents of metabolites [γ aminobutyric acid （GABA） and 5-hydroxytryptamine （5-HT）] in slow transit constipation （STC） rats. METHODS SD rats were randomly divided into blank group （10 rats） and modeling group （30 rats）， with half male and half female. The STC model was established by intragastric administration of Compound diphenoxylate tablets in the modeling group. The successfully modeled rats were randomly divided into model group， Modified shaoyao gancao decoction group [56 g/（kg·d）， calculated by crude drug] and positive control group [lactulose 2.09 g/（kg·d）]， with 10 rats in each group. Each administration group was given relevant medicine intragastrically， the blank group and model group received an equivalent volume of normal saline， once a day， for 14 consecutive days. During the experiment， the general situation of rats was observed in each group. After the last medication， the body weight was measured， and the Bristol score was used to evaluate the fecal characteristics. The fecal moisture content， intestinal propulsion rate， and the contents of GABA and 5-HT in intestinal content were detected； the diversity of intestinal flora in intestinal contents was investigated， and the correlation between the contents of GABA， 5-HT and relative abundance of microbiota was analyzed. RESULTS Compared with the model group， general conditions such as small body shape， sparse and rough fur， and slow movement were all improved in Modified shaoyao gancao decoction body weight， Bristol score， fecal moisture content，intestinal propulsion rate， 5-HT content， Chao1 index and Shannon index were increased significantly， while GABA content and Simpson index were decreased significantly （P＜0.05）. The intestinal flora of rats in the Modified shaoyao gancao decoction group could be classified as the same as the blank group， but was far from the model group； the relative abundances of Desulfobacterota， Firmicutes and Bacteroidota in this group showed a tendency of pull back， but the differences were not statistically significant compared to model group （P＞0.05）. Desulfobacterota was an intergroup differential factor （P＜0.05）. The content of GABA was negatively correlated with the relative abundance of Bacteroidota， Cyanobacteria， Patescibacteria and Actinobacteriota （P＜0.05）. The content of 5-HT was positively correlated with the relative abundance of Campilobacterota （P＜0.05）. CONCLUSIONS Modified shaoyao gancao decoction can improve the fecal properties and intestinal motility of STC rats. Its mechanism may be related to improving intestinal flora and then affecting the contents of GABA and 5-HT in intestinal contents. In addition， the contents of GABA and 5-HT may be significantly correlated with the relative abundance of specific bacterial phyla such as Bacteroidota and Campilobacterota.

BACKGROUND:Obesity negatively affects dynamic balance during walking,and crossing barriers is a more routine functional activity that requires more stability in controlling body posture. OBJECTIVE:To investigate the differences in dynamic stability between obese and average males,and to assess the balance ability of obese males using a relatively more challenging obstacle crossing. METHODS:A total of 24 male youths(12 each in the obese and normal groups)were recruited to complete the tests of walking on level ground and crossing obstacles of different heights(4 cm,11 cm,15 cm)in random order.Kinematic and dynamic data were collected using the Qualisys motion capture system and Kistler force stage.Statistical analysis was performed using two-factor(2 groups * 4 movement types)repeated measures analysis of variance. RESULTS AND CONCLUSION:The obese group had a lower step speed than the normal group(P<0.05),the proportion of the first single support period decreased and the proportion of the second double support period increased when crossing the 11 cm versus 15 cm hurdles(P<0.05).When walking on level ground,the margin of stability in the internal and external directions in the normal group was greater than that of the obese group(P<0.05).When crossing the 4 cm hurdles,the margin of stability in the obese group was less than that in the normal group(P<0.05).When crossing the 11 cm hurdles,there was no significant difference between the two groups in the anterior-posterior direction(P>0.05),while there was a significant difference in the internal-external direction(P<0.05).When crossing the 15 cm hurdles,the margin of stability in the obese group was lower than that in the normal group(P<0.05).Overall,obesity decreases the body's ability to control the body,reduces dynamic stability during crossing the barrier,and increases the risk of falls compared with the general population.In addition,compared with level ground walking,the decrease in the dynamic stability when crossing barriers is more significant in the obese group than the general population.

Objective To explore the mechanism of modified maimendong decoction (MMD) in inhibiting lung cancer metastasis from the perspective of immune regulation. Methods CTC-TJH-01 and LLC cells were intervened with different concentrations of modified maimendong decoction. The cell proliferation was detected with a CCK-8 kit, apoptosis was detected with an Annexin V-FITC/PI kit, and cell migration was detected through Transwell assays. A lung metastasis model was established through the tail vein injection of LLC cells into C57BL/6 mice, and body weight change and lung tumor metastasis in the mice were evaluated after continuous gavage intervention with MMD. HE staining, immunohistochemistry, and immunofluorescence were employed to observe the histomorphology, Ki-67 protein level, and NK and T cell levels of metastatic lesions. The levels of NK and T cells in the peripheral blood of mice were detected throughflow cytometry. Results MMD had no significant inhibitory effect on the proliferation, apoptosis, and migration of CTC-TJH-01 and LLC cells in vitro. In mice, MMD could significantly inhibit the lung metastasis of LLC cells, increase the proportion of NK and CD8⁺ T cells in peripheral blood and tumor microenvironment (P<0.05), and reduce the expression of Ki-67 protein in metastatic tumor tissues (P<0.05). Conclusion MMD may inhibit the growth of metastatic tumors by upregulating the expression levels of NK and CD8⁺ T cells in peripheral blood to promote the elimination of circulating tumor cells, and regulating the infiltration of NK and CD8⁺ T cells in the immune microenvironment of metastatic tumors, then play an antimetastatic role in lung cancer.

Animal experiments are widely used in biomedical research for safety assessment, toxicological analysis, efficacy evaluation, and mechanism exploration. In recent years, the ethical review system has become more stringent, and awareness of animal welfare has continuously increased. To promote more efficient and cost-effective drug research and development, the United States passed the Food and Drug Administration (FDA) Modernization Act 2.0 in September 2022, which removed the federal mandate requiring animal testing in preclinical drug research. In April 2025, the FDA further proposed to adopt a series of "new alternative methods" in the research and development of drugs such as monoclonal antibodies, which included artificial intelligence computing models, organoid toxicity tests, and 3D micro-physiological systems, thereby gradually phasing out traditional animal experiment models. Among these cutting-edge technologies, 3D bioprinting models are a significant alternative and complement to animal models, owing to their high biomimetic properties, reproducibility, and scalability. This review provides a comprehensive overview of advancements and applications of 3D bioprinting technology in the fields of biomedical and pharmaceutical research. It starts by detailing the essential elements of 3D bioprinting, including the selection and functional design of biomaterials, along with an explanation of the principles and characteristics of various printing strategies, highlighting the advantages in constructing complex multicellular spatial structures, regulating microenvironments, and guiding cell fate. It then discusses the typical applications of 3D bioprinting in drug research and development,including high-throughput screening of drug efficacy by constructing disease models such as tumors, infectious diseases, and rare diseases, as well as conducting drug toxicology research by building organ-specific models such as those of liver and heart. Additionally,the review examines the role of 3D bioprinting in tissue engineering, discussing its contributions to the construction of functional tissues such as bone, cartilage, skin, and blood vessels, as well as the latest progress in regeneration and replacement. Furthermore, this review analyzes the complementary advantages of 3D bioprinting models and animal models in the research of disease progression, drug mechanisms, precision medicine, drug development, and tissue regeneration, and discusses the potential and challenges of their integration in improving model accuracy and physiological relevance. In conclusion, as a cutting-edge in vitro modeling and manufacturing technology, 3D bioprinting is gradually establishing a comprehensive application system covering disease modeling, drug screening, toxicity prediction, and tissue regeneration.