BACKGROUND:Curcumin is the main active ingredient of turmeric and has significant medicinal value in anti-tumor,anti-inflammatory,antioxidant and other aspects.However,its poor water solubility,unstable chemical properties and easy decomposition lead to difficulty in extracting curcumin and low extraction yield.Therefore,it is particularly important to optimize the curcumin extraction method.OBJECTIVE:To enhance the extraction yield and utilization value of curcumin and optimize the curcumin extraction process and curcumin nanoparticle preparation process.METHODS:Curcumin was extracted from turmeric by ethanol extraction,ultrasonic extraction,ionic liquid extraction,enzyme extraction,and ionic liquid combined with ultrasonic assisted enzyme extraction.The curcumin extraction yield was detected by high performance liquid chromatography;the best extraction method was determined,and subsequent process optimization experiments were carried out.The curcumin extraction yield was the response value with the type of ionic liquid,reaction temperature,ultrasonic time,liquid-to-solid ratio,ionic liquid concentration,and enzyme-drug mass ratio as parameters.The optimal production process of ionic liquid combined with ultrasonic assisted enzyme extraction was determined by single factor combined response surface experiment.The optimal process for preparing curcumin nanoparticles by ionic crosslinking method was determined by single factor combined response surface experiment with acetic acid concentration,chitosan to sodium tripolyphosphate mass ratio,stirring rate,curcumin mass concentration,sodium tripolyphosphate mass concentration,and chitosan mass concentration as parameters,and drug encapsulation efficiency as response value.Curcumin nanoparticles were prepared under the optimal process,and the particle size,polydispersity index,Zata potential value,drug loading,stability,hemolysis rate,and antioxidant capacity in vivo and in vitro of the nanoparticles were detected.RESULTS AND CONCLUSION:(1)Among the five extraction methods,the curcumin yield of ionic liquid combined with ultrasound-assisted enzyme extraction was the highest,and this method was selected as the curcumin extraction method for subsequent experiments.The results of single factor combined response surface experiment showed that the optimal process for curcumin extraction was:ionic liquid selected 1-hexyl-3-methylimidazolium chloride,reaction temperature 55 ℃,liquid-to-solid ratio 40 mL/g,ultrasound time 57 minutes,ionic liquid concentration 57％,enzyme-drug mass ratio 3.5:10,and the obtained turmeric extraction yield was 3.10％.The optimal preparation process of curcumin nanoparticles was:glacial acetic acid concentration 0.5％,chitosan and sodium tripolyphosphate mass ratio 5.0:1,stirring speed 150 r/min,curcumin mass concentration 2.23 mg/mL,sodium tripolyphosphate mass concentration 1.45 mg/mL,chitosan mass concentration 3.63 mg/mL,and the obtained drug encapsulation efficiency was 90.61％.(2)The drug loading of curcumin nanoparticles was(14.49±0.23)％,the average particle size was(76.95±1.65)nm,the polydispersity coefficient was 0.15±0.02,and the Zata potential value was(32.37±1.46)mV.The curcumin nanoparticles had good stability and blood compatibility,did not induce hemolysis,and had stronger antioxidant capacity in vivo and in vitro than free curcumin.(3)The results show that the process optimization not only solves the problems of low extraction yield,poor solubility,and low bioavailability of curcumin,but also enhances its antioxidant activity in vivo and in vitro.

BACKGROUND:Curcumin is the main active ingredient of turmeric and has significant medicinal value in anti-tumor,anti-inflammatory,antioxidant and other aspects.However,its poor water solubility,unstable chemical properties and easy decomposition lead to difficulty in extracting curcumin and low extraction yield.Therefore,it is particularly important to optimize the curcumin extraction method.OBJECTIVE:To enhance the extraction yield and utilization value of curcumin and optimize the curcumin extraction process and curcumin nanoparticle preparation process.METHODS:Curcumin was extracted from turmeric by ethanol extraction,ultrasonic extraction,ionic liquid extraction,enzyme extraction,and ionic liquid combined with ultrasonic assisted enzyme extraction.The curcumin extraction yield was detected by high performance liquid chromatography;the best extraction method was determined,and subsequent process optimization experiments were carried out.The curcumin extraction yield was the response value with the type of ionic liquid,reaction temperature,ultrasonic time,liquid-to-solid ratio,ionic liquid concentration,and enzyme-drug mass ratio as parameters.The optimal production process of ionic liquid combined with ultrasonic assisted enzyme extraction was determined by single factor combined response surface experiment.The optimal process for preparing curcumin nanoparticles by ionic crosslinking method was determined by single factor combined response surface experiment with acetic acid concentration,chitosan to sodium tripolyphosphate mass ratio,stirring rate,curcumin mass concentration,sodium tripolyphosphate mass concentration,and chitosan mass concentration as parameters,and drug encapsulation efficiency as response value.Curcumin nanoparticles were prepared under the optimal process,and the particle size,polydispersity index,Zata potential value,drug loading,stability,hemolysis rate,and antioxidant capacity in vivo and in vitro of the nanoparticles were detected.RESULTS AND CONCLUSION:(1)Among the five extraction methods,the curcumin yield of ionic liquid combined with ultrasound-assisted enzyme extraction was the highest,and this method was selected as the curcumin extraction method for subsequent experiments.The results of single factor combined response surface experiment showed that the optimal process for curcumin extraction was:ionic liquid selected 1-hexyl-3-methylimidazolium chloride,reaction temperature 55 ℃,liquid-to-solid ratio 40 mL/g,ultrasound time 57 minutes,ionic liquid concentration 57％,enzyme-drug mass ratio 3.5:10,and the obtained turmeric extraction yield was 3.10％.The optimal preparation process of curcumin nanoparticles was:glacial acetic acid concentration 0.5％,chitosan and sodium tripolyphosphate mass ratio 5.0:1,stirring speed 150 r/min,curcumin mass concentration 2.23 mg/mL,sodium tripolyphosphate mass concentration 1.45 mg/mL,chitosan mass concentration 3.63 mg/mL,and the obtained drug encapsulation efficiency was 90.61％.(2)The drug loading of curcumin nanoparticles was(14.49±0.23)％,the average particle size was(76.95±1.65)nm,the polydispersity coefficient was 0.15±0.02,and the Zata potential value was(32.37±1.46)mV.The curcumin nanoparticles had good stability and blood compatibility,did not induce hemolysis,and had stronger antioxidant capacity in vivo and in vitro than free curcumin.(3)The results show that the process optimization not only solves the problems of low extraction yield,poor solubility,and low bioavailability of curcumin,but also enhances its antioxidant activity in vivo and in vitro.

Objective: To examine the relationship between self-management behaviors and time perspective among patients with comorbid diabetes, so as to provide the evidence for improving self-management behaviors among patients with comorbid diabetes. Methods: The patients with comorbid diabetes who were registered in the chronic disease health management system of Minhang District, Shanghai Municipality in 2021, followed up regularly, and lived in Meilong Town were recruited. Demographic information and family history of diabetes were collected through questionnaire surveys. Time perspective and self-management behaviors were assessed using the Zimbardo Time Perspective Inventory and Diabetes Self-Management Behavior Scale, respectively. The relationship between self-management behaviors and time perspective was analyzed using a multivariable ordinal logistic regression model. Results: A total of 907 patients with comorbid diabetes were enrolled, including 472 males (52.04%) and 435 females (47.96%). There were 652 cases aged 65 years and above, accounting for 71.89%. In terms of the types of time perspective, 280 patients were future-oriented (30.87%), 236 were balanced (26.02%), 162 were sensation-seeking (17.86%), 123 were fatalistic (13.56%), and 106 were negative (11.69%). In terms of the self-management behaviors, 46 patients were good (5.07%), 643 were moderate (70.89%), and 218 were poor (24.04%). Multivariable ordinal logistic regression analysis showed that after adjusting for age, gender, educational level, marital status, occupation status, monthly income, and family history of diabetes, the patients with comorbid diabetes who had a future-oriented time perspective had better self-management behaviors (OR=1.874, 95%CI: 1.204-2.915). Conclusion The self-management behaviors among patients with comorbid diabetes are moderate to poor, and patients with a future-oriented time perspective can better engage in self-management behaviors.

Objective To investigate the prevalence and the risk factors of heart disease(HD)in adults aged ≥80 years old in China based on the data from the 8th round of the Chinese Longitudinal Healthy Longevity Survey(CLHLS).Methods A total of 7 675 adults aged ≥80 years old were enrolled from the 8th round of CLHLS dataset.Chi-square tests were employed to examine associations between cardiovascular disease and demographic characteristics,socioeconomic status,social support,lifestyle factors,and health indicators.Logistic regression models were developed to analyze significant predictors of heart disease in the elderly.Results The prevalence of heart disease was 16％(n=1 228)in 7 675 elderly people.Aged 90-99 years old(odds ratio[OR]=0.816),≥100 years old(OR=0.641),female(OR=0.833),and low body mass index(BMI)(＜18.5 kg/m2,OR=0.778)were the protective factors for cardiovascular disease in the elderly;and high BMI(24.0 to 27.9 kg/m2,OR=1.209),rural residence(OR=2.384),health examination(OR=1.164),dysfunction of daily living activities(OR=1.401),hypertension(OR=2.143),diabetes mellitus(OR=1.719),and history of cerebrovascular accident(OR=2.080)were risk factors.Conclusion Male,overweight,rural residence,health examination,dysfunction of daily living activities,hypertension,diabetes mellitus,and a history of cerebrovascular accident are the risk factors for heart disease in the elderly.

Objective:To investigate the effects of Zhibai Dihuang Pills on neurons of cognitive dysfunction in D-galactose (D-gal) model mice.Methods:Totally 60 male mice were divided into four groups using a random number table method: control group, model group, donepezil group, and Zhibai Dihuang Pills group, with 15 mice in each group. Except for the control group, the other groups were subcutaneously injected with D-galactose solution at a dosage of 125 mg/kg once a day for 8 weeks to prepare the aging model. Mice in the donepezil group were intragastrically administered donepezil solution at a dosage of 0.65 mg/kg, and those in the Zhibai Dihuang Pills group were intragastrically administered Zhibai Dihuang Pills solution at a dosage of 1.56 g/kg. The control group was intragastrically administered an equal volume of physiological saline once a day for 8 weeks. The object recognition test and Morris Water Maze were used to assess object recognition memory and spatial learning memory abilities of mice in each group, respectively. Hematoxylin-Eosin (HE) staining and Nissl staining were employed to observe the morphology of neurons in the hippocampal region; Golgi staining was used to observe neuronal dendritic spines; Western Blot was used to detect the protein expression levels of PI3K, p-Akt/Akt, glycogen synthase kinase 3β (GSK3β), postsynaptic density protein-95 (PSD-95), and synaptophysin (SYP) in the hippocampus region; RT-qPCR was performed to detect mRNA expression of PI3K, Akt and GSK3β in the hippocampus region.Results:Compared with the model group, the recognition index in both the donepezil group and the Zhibai Dihuang Pills group increased ( P<0.05), the escape latency was shortened ( P<0.05), the platform crossings times and the target quadrant dwell time increased ( P<0.05), the number of nerve cells in the hippocampal region increased, arranged closely, the number of Nissl bodies increased, the morphology returned to normal, and the density of dendritic spines increased; the protein expressions of PI3K, PSD-95, and SYP in the hippocampal region and the ratio of p-Akt/Akt increased ( P<0.01), the mRNA level of PI3K increased ( P<0.01 or P<0.05), and the protein and mRNA levels of GSK3β decreased ( P<0.01 or P<0.05). Conclusion:Zhibai Dihuang Pills can improve the learning and memory ability and rescue neuronal damage in D-gal model mice, and the mechanism may be related to the activation of PI3K/Akt pathway and the restoration of synaptic connections.

Objective: To investigate the association between screen time (ST) during leisure time and anxiety-depression symptoms among middle school students, so as to provide a basis for formulating relevant intervention measures. Methods: From November to December 2023, a stratified cluster random sampling method was used to select 2 542 students from junior and senior high school in Taiyuan City. A self designed questionnaire, the Generalized Anxiety Disorder Scale (GAD-7), and the Patient Health Questionnaire (PHQ-9) were used to investigate ST and anxiety/depression symptoms among middle school students. The Logistic regression model was used to explore the association of ST with symptoms of anxiety and depression, as well as with anxiety and depression comorbiditles (CAD). Results: The detection rates of anxiety symptoms, depression symptoms, and CAD were 13.69%, 15.77%, and 10.11%, respectively. The median ST was 2.00 h/d [interquartile range ( IQR =2.38) for weekly averages], with 0.33 h/d ( IQR =1.67) on work days and 5.00 h/d ( IQR=5.50) on rest days. Logistic regression analysis indicated that the total ST of mobile phones during rest days ( OR =1.07, 1.10, 1.11) and the averages ST of mobile phones over a week ( OR = 1.20 , 1.22, 1.29), as well as the total ST of all screen types during rest days ( OR =1.04, 1.04, 1.05) and the averages ST of all screen types over a week ( OR =1.08, 1.09, 1.21) were positively correlated with anxiety symptoms, depression symptoms, and CAD (all P <0.01). Conclusions Among middle school students in Taiyuan City, screen time is positively correlated with symptoms of anxiety or depression and the comorbidity of anxiety and depression, especially smartphone screen time and weekend screen use. Therefore, measures should be implemented to reduce unnecessary screen time among middle school students, especially the use of mobile phones, in order to mitigate the occurrence of anxiety and depression.

Objective To observe the effects of Baishile Capsules on neonatal neuronal activity and synaptic plasticity in hippocampus of depression model rats;To explore its mechanism of antidepressant.Methods Totally 32 SD rats were randomly divided into blank group,model group,fluoxetine group(5.4 mg/kg)and Baishile Capsules group(2.88 g/kg),with 8 rats in each group.The depression rat model was established by chronic unpredictable mild stress and single cage feeding,and drugs were administered at the same time as modeling for 21 consecutive days.Forced swimming test and open field test were used to evaluate the depressive-like behavior of rats,Golgi staining was used to observe the synaptic morphology of hippocampal neurons,and immunofluorescence was used to detect the positive expressions of BrdU/GABA-B,BrdU/c-fos,BrdU/GAP-43,BrdU/MAP-2 and CXCR4 in hippocampal tissue.Results Compared with the blank group,the immobility time of forced swimming experiment in the model group increased,and the horizontal and vertical time of open field experiment decreased significantly(P<0.01);the hippocampal neurons dendrites and dendritic spines atrophied,the density decreased,the branches became shorter,synaptic structure becomes blurred,and the longest and total lengths of synaptic branches significantly decreased(P<0.01);the positive expressions of GABA-B,c-fos,GAP-43 and MAP-2 proteins in hippocampal neonatal neurons decreased(P<0.05,P<0.01),and the expression of CXCR4 in hippocampal tissue decreased significantly(P<0.01).Compared with the model group,the immobility time of the forced swimming experiment in fluoxetine group and Baishile Capsules group significantly reduced,and the horizontal and vertical activity time of the open field experiment significantly increased(P<0.01);the number of dendritic spines in hippocampal neurons increased,synaptic damage improved,and the length of the longest and total branches of synapses significantly increased(P<0.01);the positive expressions of GABA-B,c-fos,GAP-43 and MAP-2 in hippocampal neonatal neurons significantly increased(P<0.05,P<0.01),and the positive expression of CXCR4 in hippocampal tissue significantly increased(P<0.01).Conclusion Baishile Capsules can regulate hippocampal synaptic plasticity and nerve regeneration by improving the activity and function of hippocampal neonatal neurons in depression model rats,exerting antidepressant effects.

OBJECTIVES: To explore the mechanism of Pingchuanning Formula (PCN) for inhibiting airway inflammation in rats with asthmatic cold syndrome. METHODS: A total of 105 SD rats were randomized equally into 7 groups, including a control group, an asthmatic cold syndrome model group, 3 PCN treatment groups at high, medium and low doses, a Guilong Kechuanning (GLCKN) treatment group, and a dexamethasone (DEX) treatment group. In all but the control rats, asthma cold syndrome models were established and daily gavage of saline, PCN, GLCKN or DEX was administered 29 days after the start of modeling. The changes in general condition, lung function and lung histopathology of the rats were observed, and inflammatory factors in the alveolar lavage fluid (BALF), oxidative stress, lung tissue ultrastructure, cytokine levels, and expressions of the genes related to the HMGB1/Beclin-1 axis and autophagy were analyzed. RESULTS: The rat models had obvious manifestations of asthmatic cold syndrome with significantly decreased body mass, food intake, and water intake, reduced FEV_0.3, FVC, and FEV_0.3/FVC, obvious inflammatory cell infiltration in the lung tissue, and increased alveolar inflammation score and counts of neutrophils, eosinophils, lymphocytes, macrophages, and leukocytes in the BALF. The rat models also had significantly increased MDA level and decreased SOD level and exhibited obvious ultrastructural changes in the lung tissues, where the expressions of HMGB1, Beclin-1, ATG5, TNF-α, IL-6,IL-1β, and IL-13 and the LC3II/I ratio were increased, while the levels of Bcl-2 and IFN-γ were decreased. PCN treatment significantly improved these pathological changes in the rat models, and its therapeutic effect was better than that of GLKCN and similar to that of DEX. CONCLUSIONS PCN can effectively alleviate airway inflammation in rat models of asthmatic cold syndrome possibly by modulating the HMGB1/Beclin-1 signaling axis to suppress cell autophagy, thereby attenuating airway inflammatory damages.
Animals ; Rats ; Autophagy/drug effects* ; Rats, Sprague-Dawley ; Asthma/pathology* ; Beclin-1 ; HMGB1 Protein/metabolism* ; Drugs, Chinese Herbal/therapeutic use* ; Disease Models, Animal ; Male ; Lung/pathology* ; Inflammation

The accurate segmentation of breast ultrasound images is an important precondition for the lesion determination. The existing segmentation approaches embrace massive parameters, sluggish inference speed, and huge memory consumption. To tackle this problem, we propose T ²KD Attention U-Net (dual-Teacher Knowledge Distillation Attention U-Net), a lightweight semantic segmentation method combined double-path joint distillation in breast ultrasound images. Primarily, we designed two teacher models to learn the fine-grained features from each class of images according to different feature representation and semantic information of benign and malignant breast lesions. Then we leveraged the joint distillation to train a lightweight student model. Finally, we constructed a novel weight balance loss to focus on the semantic feature of small objection, solving the unbalance problem of tumor and background. Specifically, the extensive experiments conducted on Dataset BUSI and Dataset B demonstrated that the T ²KD Attention U-Net outperformed various knowledge distillation counterparts. Concretely, the accuracy, recall, precision, Dice, and mIoU of proposed method were 95.26%, 86.23%, 85.09%, 83.59%and 77.78% on Dataset BUSI, respectively. And these performance indexes were 97.95%, 92.80%, 88.33%, 88.40% and 82.42% on Dataset B, respectively. Compared with other models, the performance of this model was significantly improved. Meanwhile, compared with the teacher model, the number, size, and complexity of student model were significantly reduced (2.2×10 ⁶ vs. 106.1×10 ⁶, 8.4 MB vs. 414 MB, 16.59 GFLOPs vs. 205.98 GFLOPs, respectively). Indeedy, the proposed model guarantees the performances while greatly decreasing the amount of computation, which provides a new method for the deployment of clinical medical scenarios.
Humans ; Breast Neoplasms/diagnostic imaging* ; Female ; Ultrasonography, Mammary/methods* ; Image Processing, Computer-Assisted/methods* ; Algorithms ; Neural Networks, Computer ; Breast/diagnostic imaging*

Objective To investigate the effect and mechanism of Efferocytosis Relatived LncRNA (EFRL) on homocysteine-induced atherosclerosis in macrophage efferocytosis. Methods RAW264.7 cells were cultured in vitro, and the Control group (0 μmol/L Hcy) and Hcy intervention group (100 μmol/L Hcy) were set up. After GapmeR transfection of macrophages with Hcy intervention, EFRL knockdown negative control group (Hcy combined with LNA-NC) and EFRL knockdown group (Hcy combined with LNA-EFRL) were set up. High-throughput sequencing was applied for different expression of LncRNA MSTRG. 88917.16 (EFRL), UCSC was used to analyze its conservation, CPC and CPAT were used to analyze its ability to encode proteins, and GO and KEGG were used to analyze related biological functions. The localization of LncRNA EFRL in macrophages was analyzed by nucleoplasmic separation and RNA-FISH. Quantitative real-time PCR was used to detect the expression levels of LncRNA EFRL and its target gene SPAST in Hcy-treated macrophages. The apoptosis rate of Jurkat cells induced by UV was detected by flow cytometry. In vitro efferocytosis assay combined with immunofluorescence technique was used to analyze macrophage efferocytosis. ELISA was used to detect the levels of interleukin 1β(IL-1β) and IL-18. Results The new LncRNA MSTRG.88917.16 was identified and named EFRL(Efferocytosis Relatived LncRNA). UCSC, CPC and CPAT analyses showed that LncEFRL is highly conserved and does not have the ability to encode proteins. GO and KEGG analyses suggested that LncEFRL may be involved in macrophage efferocytosis. LncRNA EFRL was localized in the nucleus of macrophages as determined by nucleoplasmic separation and RNA-FISH. In comparison to the Control group, the expression levels of LncRNA EFRL and its target gene SPAST in the Hcy group were increased. In comparison to the Control group (0 min), the apoptosis rate of the experimental group (15, 30 min) Annexin V is more than 85%. Compared with Hcy combined with LNA-NC group, Hcy combined with LNA-EFRL group had enhanced macrophage efferocytosis and reduced levels of inflammatory factors. Compared with Hcy combined with LNA-NC group, the expression level of SPAST in Hcy combined with LNA-EFRL group was decreased. Conclusion Inhibition of EFRL expression can alleviate the process of Hcy inhibiting macrophage efferocytosis, and the mechanism is related to the regulation of the downstream target gene SPAST by EFRL.
RNA, Long Noncoding/physiology* ; Animals ; Homocysteine ; Mice ; Macrophages/drug effects* ; Humans ; RAW 264.7 Cells ; Atherosclerosis/chemically induced* ; Apoptosis/genetics* ; Phagocytosis/genetics* ; Jurkat Cells ; Interleukin-1beta/genetics* ; Efferocytosis