1.Establishment and application of a rapid high-throughput detection method for Huanglongbing.
Qin YUAN ; Zhi-Peng LI ; Tie-Lin WANG ; Ting DONG ; Yu-Wen YANG ; Wei GUAN ; Ting-Chang ZHAO
China Journal of Chinese Materia Medica 2025;50(7):1735-1740
The dried mature peel of Citrus reticulata, a plant in the Rutaceae family and its cultivated varieties, is a commonly used Chinese medicinal material known as Chenpi(Citri Reticulatae Pericarpium). It is rich in nutritional components and medicinal value, with pharmacological effects including relieving cough and eliminating phlegm, strengthening the spleen and drying dampness, protecting the liver and benefiting the stomach, tonifying Qi, and calming the mind. Huanglongbing(HLB), also known as Citrus Huanglongbing, is a destructive disease in citrus production that seriously threatens the development of the citrus industry. HLB causes symptoms such as the inability of Rutaceae plants to produce mature fruit, gradual weakening of the tree, and eventual death, posing a significant threat to the yield and quality of Chenpi. Due to the uneven distribution of the HLB pathogen in infected plants, accurate detection of the pathogen requires the collection of a large number of plant samples. Current sample pretreatment methods, such as traditional extraction methods and commercial extraction kits, are time-consuming and involve multiple steps, which significantly increase the difficulty and workload of HLB diagnosis and have become a bottleneck in HLB detection. In this study, a rapid high-throughput detection method combining alkali lysis and TaqMan qPCR was developed. This method allows the pretreatment of multiple samples within 5 min, and the entire detection process can be completed within 45 min, with a detection limit of 6.67 fg·μL~(-1). The alkali lysis method and commercial kits were used for parallel detection of field-collected citrus samples, and the results showed no significant difference. The sample pretreatment method established in this study is characterized by low cost, simplicity, and high efficiency. Combined with TaqMan qPCR, it can provide technical support for early and on-site diagnosis of HLB. This method is of great significance for disease prevention and control in the citrus industry and is expected to help improve the yield and quality of citrus medicinal materials.
Citrus/microbiology*
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Plant Diseases/microbiology*
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Rhizobiaceae/physiology*
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High-Throughput Screening Assays/methods*
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Liberibacter/physiology*
2.Buzhong Yiqi Decoction alleviates immune injury of autoimmune thyroiditis in NOD.H-2~(h4)mice via c GAS-STING signaling pathway.
Yi-Ran CHEN ; Lan-Ting WANG ; Qing-Yang LIU ; Zhao-Han ZHAI ; Shou-Xin JU ; Xue-Ying CHEN ; Zi-Yu LIU ; Xiao YANG ; Tian-Shu GAO ; Zhi-Min WANG
China Journal of Chinese Materia Medica 2025;50(7):1872-1880
This study aims to explore the effects of Buzhong Yiqi Decoction(BYD) on the cyclic guanosine monophosphate-adenosine monophosphate synthase(cGAS)-stimulator of interferon genes(STING) signaling pathway in the mouse model of autoimmune thyroiditis(AIT) and the mechanism of BYD in alleviating the immune injury. Forty-eight NOD.H-2~(h4) mice were assigned into normal, model, low-, medium-, and high-dose BYD, and selenium yeast tablets groups(n=8). Mice of 8 weeks old were treated with 0.05% sodium iodide solution for 8 weeks for the modeling of AIT and then administrated with corresponding drugs by gavage for 8 weeks before sampling. High performance liquid chromatography was employed to measure the astragaloside Ⅳ content in BYD. Hematoxylin-eosin staining was employed to observe the pathological changes in the mouse thyroid tissue. Enzyme-linked immunosorbent assay was employed to measure the serum levels of thyroid peroxidase antibody(TPO-Ab), thyroglobulin antibody(TgAb), and interferon-γ(IFN-γ). Flow cytometry was employed to detect the distribution of T cell subsets in the spleen. The immunohistochemical method was used to detect the expression of cGAS, STING, TANK-binding kinase 1(TBK1), and interferon regulatory factor 3(IRF3). Real-time PCR and Western blot were employed to determine the mRNA and protein levels, respectively, of markers related to the cGAS-STING signaling pathway in the thyroid tissue. The results showed that the content of astragaloside Ⅳ in BYD was(7.06±0.08) mg·mL~(-1). Compared with the normal group, the model group showed disrupted structures of thyroid follicular epithelial cells, massive infiltration of lymphocytes, and elevated levels of TgAb and TPO-Ab. Compared with the model group, the four treatment groups showed intact epithelial cells, reduced lymphocyte infiltration, and lowered levels of TgAb and TPO-Ab. Compared with the normal group, the model group showed increases in the proportions of Th1 and Th17 cells, a decrease in the proportion of Th2 cells, and an increase in the IFN-γ level. Compared with the model group, the four treatment groups presented decreased proportions of Th1 and Th17 cells and lowered levels of IFN-γ, and the medium-dose BYD group showed an increase in the proportion of Th2 cells. Compared with the normal group, the modeling up-regulated the mRNA levels of cGAS, STING, TBK1, and IRF3 and the protein levels of cGAS, p-STING, p-TBK1, and p-IRF3. Compared with the model group, the four treatment groups showed reduced levels of cGAS, STING, TBK1, and IRF3-positive products, down-regulated mRNA levels of cGAS, STING, and TBK1, and down-regulated protein levels of cGAS and p-STING. The high-dose BYD group showed down-regulations in the mRNA level of IRF3 and the protein levels of p-TBK1 and p-IRF3. The above results indicate that BYD can repair the imbalance of T cell subsets, alleviate immune injury, and reduce thyroid lymphocyte infiltration in AIT mice by inhibiting the cGAS-STING signaling pathway.
Animals
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Drugs, Chinese Herbal/administration & dosage*
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Signal Transduction/drug effects*
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Thyroiditis, Autoimmune/metabolism*
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Mice
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Membrane Proteins/metabolism*
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Mice, Inbred NOD
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Humans
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Female
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Nucleotidyltransferases/metabolism*
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Male
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Disease Models, Animal
3.Two new taraxerane triterpenoids from mastic.
Zhi-Qiang ZHAO ; Xue-Rui AN ; Tian-Zhi LI ; Ting HE ; Hao-Kun HOU ; Wei LIU ; Tao YUAN
China Journal of Chinese Materia Medica 2025;50(13):3723-3743
Three taraxerane nortriterpenoids were isolated from mastic by using various modern chromatographic separation techniques. They were identified as(5R,8R,9R,10S,11S,12R,13S,17R,18R)-28-norlupa-11,12-epoxy-14-taraxerene-3,16-dione(1),(5R,8R,9R,10S,11S,12R,13S,17S,18S)-17-hydroxy-28-norlupa-11,12-epoxy-14-taraxerene-3-one(2), and(5R,8R,9R,10R,11S,12R,13R,14S,17S,18S)-14,17-epoxy-28-norlupa-11,12-oxidotaraxerone(3) through the high-resolution electrospray ionization mass spectrometry(HR-ESI-MS), infrared(IR), ultraviolet(UV), nuclear magnetic resonance(NMR), and single-crystal X-ray diffraction techniques as well as comparison with literature data. Compounds 1-3 were C-28 nortriterpenoids and isolated from mastic for the first time, and compounds 1-2 were new ones. In the model for RAW264.7 cell anti-inflammation induced by lipopolysaccharide(LPS), compound 1 demonstrates an inhibitory effect on nitric oxide(NO) [IC_(50)=(13.38±0.68) μmol·L~(-1)], comparable to the activity of the positive control dexamethasone [IC_(50)=(14.59±1.49) μmol·L~(-1)]. Compounds 2 and 3 exhibit weaker inhibitory effects, with IC_(50) values of(24.17±2.56) and(22.25±2.84) μmol·L~(-1), respectively.
Animals
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Mice
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Triterpenes/isolation & purification*
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Drugs, Chinese Herbal/isolation & purification*
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Mastic Resin/chemistry*
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Nitric Oxide
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Molecular Structure
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Macrophages/immunology*
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RAW 264.7 Cells
4.Clinical application of single-balloon and double-balloon enteroscopy in pediatric small bowel diseases: a retrospective study of 576 cases.
Can-Lin LI ; Jie-Yu YOU ; Yan-Hong LUO ; Hong-Juan OU-YANG ; Li LIU ; Wen-Ting ZHANG ; Jia-Qi DUAN ; Na JIANG ; Mei-Zheng ZHAN ; Chen-Xi LIU ; Juan ZHOU ; Ling-Zhi YUAN ; Hong-Mei ZHAO
Chinese Journal of Contemporary Pediatrics 2025;27(7):822-828
OBJECTIVES:
To evaluate the effectiveness of single-balloon and double-balloon enteroscopy in diagnosing pediatric small bowel diseases and assess the diagnostic efficacy of computed tomography enterography (CTE) for small bowel diseases using enteroscopy as the reference standard.
METHODS:
Clinical data from 576 children who underwent enteroscopy at Hunan Children's Hospital between January 2017 and December 2023 were retrospectively collected. The children were categorized based on enteroscopy type into the single-balloon enteroscopy (SBE) group (n=457) and double-balloon enteroscopy (DBE) group (n=119), and the clinical data were compared between the two groups. The sensitivity and specificity of CTE for diagnosing small bowel diseases were evaluated using enteroscopy results as the standard.
RESULTS:
Among the 576 children, small bowel lesions were detected by enteroscopy in 274 children (47.6%).There was no significant difference in lesion detection rates or complication rates between the SBE and DBE groups (P>0.05), but the DBE group had deeper insertion, longer procedure time, and higher complete small bowel examination rate (P<0.05). The complication rate during enteroscopy was 4.3% (25/576), with 18 cases (3.1%) of mild complications and 7 cases (1.2%) of severe complications, which improved with symptomatic treatment, surgical, or endoscopic intervention. Among the 412 children who underwent CTE, the sensitivity and specificity for diagnosing small bowel diseases were 44.4% and 71.3%, respectively.
CONCLUSIONS
SBE and DBE have similar diagnostic efficacy for pediatric small bowel diseases, but DBE is preferred for suspected deep small bowel lesions and comprehensive small bowel examination. Enteroscopy in children demonstrates relatively good overall safety. CTE demonstrates relatively low sensitivity but comparatively high specificity for diagnosing small bowel diseases.
Retrospective Studies
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Treatment Outcome
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Double-Balloon Enteroscopy/statistics & numerical data*
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Single-Balloon Enteroscopy/statistics & numerical data*
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Humans
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Male
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Female
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Child
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Operative Time
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Tomography, X-Ray Computed/statistics & numerical data*
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Sensitivity and Specificity
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Intestine, Small/surgery*
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Intestinal Diseases/surgery*
5.Mechanism of Regulating MK2 to Improve Bone Marrow Inflammatory Damage after Hematopoietic Stem Cell Transplantation.
Zhao-Hui WANG ; Bo LONG ; Yu-Han WANG ; Zhi-Ting LIU ; Zi-Jie XU ; Shuang DING
Journal of Experimental Hematology 2025;33(5):1453-1460
OBJECTIVE:
To investigate the role of MK2 inhibitor MMI-0100 on inflammatory response after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and related mechanisms.
METHODS:
An allo-HSCT mouse model was established. Recipient rats were randomly divided into BMT+NaCl group and BMT+MMI-0100 group, and were injected with NaCl and MMI-0100 every day after transplantation, respectively. Samples of the two groups were collected on d 7 and 14, femur paraffin sections were stained with HE, and pathological changes in the bone marrow cavity were observed under the light microscope. The gene and protein expression levels of pro-inflammatory cytokines IL-1β and IL-18 were detected by qPCR and Western blot. Macrophage typing was detected by flow cytometry. The expression levels of NLRP3 and Caspase-1 were detected by Western blot.
RESULTS:
Inflammatory cell infiltration in the bone marrow cavity was significantly reduced in the BMT+MMI-0100 group. Western blot results showed that the protein expression levels of IL-1β and IL-18 in the BMT+MMI-0100 group were decreased compared to the BMT+NaCl group on day 7 and day 14 (all P <0.01). The qPCR results showed that compared to the BMT+NaCl group, the IL-18 gene expression levels in the BMT+MMI-0100 group were significantly reduced on day 7 and day 14 (both P <0.01). In the BMT+MMI-0100 group, the expression level of IL-1β gene decreased on day 7 (P <0.05), but increased and was higher than that in the BMT+NaCl group on day 14 (P <0.05). Flow cytometry results showed that the expression of M1 macrophages and M1/M2 ratio decreased in the BMT+MMI-0100 group compared to BMT+NaCl group (all P <0.05). Western blot results showed that the protein expression levels of NLRP3 and Caspase-1 in the BMT+MMI-0100 group were lower than those in the BMT+NaCl group (all P <0.05).
CONCLUSION
MMI-0100 can ameliorate bone marrow inflammatory injury after allo-HSCT and may act by reducing NLRP3 expression to promote M2 polarization.
Animals
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Interleukin-1beta/metabolism*
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Rats
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Interleukin-18/metabolism*
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Hematopoietic Stem Cell Transplantation/adverse effects*
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Mice
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NLR Family, Pyrin Domain-Containing 3 Protein/metabolism*
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Inflammation
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Bone Marrow/pathology*
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Protein Serine-Threonine Kinases/metabolism*
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Intracellular Signaling Peptides and Proteins/antagonists & inhibitors*
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Caspase 1/metabolism*
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Macrophages
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Transplantation, Homologous
6.Histaminergic Innervation of the Ventral Anterior Thalamic Nucleus Alleviates Motor Deficits in a 6-OHDA-Induced Rat Model of Parkinson's Disease.
Han-Ting XU ; Xiao-Ya XI ; Shuang ZHOU ; Yun-Yong XIE ; Zhi-San CUI ; Bei-Bei ZHANG ; Shu-Tao XIE ; Hong-Zhao LI ; Qi-Peng ZHANG ; Yang PAN ; Xiao-Yang ZHANG ; Jing-Ning ZHU
Neuroscience Bulletin 2025;41(4):551-568
The ventral anterior (VA) nucleus of the thalamus is a major target of the basal ganglia and is closely associated with the pathogenesis of Parkinson's disease (PD). Notably, the VA receives direct innervation from the hypothalamic histaminergic system. However, its role in PD remains unknown. Here, we assessed the contribution of histamine to VA neuronal activity and PD motor deficits. Functional magnetic resonance imaging showed reduced VA activity in PD patients. Optogenetic activation of VA neurons or histaminergic afferents significantly alleviated motor deficits in 6-OHDA-induced PD rats. Furthermore, histamine excited VA neurons via H1 and H2 receptors and their coupled hyperpolarization-activated cyclic nucleotide-gated channels, inward-rectifier K+ channels, or Ca2+-activated K+ channels. These results demonstrate that histaminergic afferents actively compensate for Parkinsonian motor deficits by biasing VA activity. These findings suggest that targeting VA histamine receptors and downstream ion channels may be a potential therapeutic strategy for PD motor dysfunction.
Animals
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Histamine/metabolism*
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Male
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Oxidopamine/toxicity*
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Rats
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Ventral Thalamic Nuclei/physiopathology*
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Rats, Sprague-Dawley
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Disease Models, Animal
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Parkinson Disease/metabolism*
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Neurons/physiology*
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Humans
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Optogenetics
7.Curcumin improving behavioral deficits in Parkinson's disease mice via modulation of gut microbiota
Wen-Hui LI ; Zhi-Hong ZHAO ; Li-Juan WANG ; Jin-Jing HE ; Yu-Ting LIU ; Qiu-Qin HAN
Acta Anatomica Sinica 2025;56(2):143-149
Objective To explore the mechanism by which curcumin improves behavioral deficits in mice with Parkinson's disease(PD)through fecal microbiota transplantation.Methods A subacute model of PD in mice was induced by intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP).Fecal microbiota from both the model group and the curcumin(Cur)-treated group(80 m g/kg)were collected and analyzed.The experiment involving fecal microbiota transplantation was structured into four distinct groups,fecal microbiota solvent transplantation group(FMTcon),model fecal microbiota transplantation group(FMTmodel),MPTP-induced model group(model),and model group subjected to fecal microbiota transplantation following curcumin treatment(model+FMTCur).The motor skills of the mice were assessed by using rod rotation,pole climbing experiment,and open field tests.Immunofluorescence techniques were employed to observe the expression tyrosine hydroxylase(TH)-positive neurons in the substantia nigra of the brain.Additionally,the gene expression of tumor necrosis factor-α(TNF-α)in the midbrain of mice was analyzed,alongside the protein expression of nuclear factor-κB(NF-κB)and nucleotide binding oligomerization domain-like receptor protein 3(NLRP3).Results The subacute PD animal model in mice was successfully established,and fecal microbiota were separated and gathered.The model group exhibited significant motor impairment,as evidenced by a shortened rod rotation time(P<0.05),prolonged pole climbing time(P<0.05),significantly reduced total movement distance within the open field(P<0.001),and decreased time spent in the central zone(P<0.01).The relative expression level of TH+neurons in the substantia nigra was significantly reduced(P<0.01).Moreover,mRNA expression of TNF-α in the midbrain increased significantly(P<0.01),along with significant elevations in protein expression of NF-κB(P<0.001),phosphorylated NF-κB(p-NF-κB)(P<0.01),NLRP3(P<0.001),and Caspase-1(P<0.01).The transplanted model microbial group(FMTmodel)also exhibited motor impairment,manifested by a trend of shortened rod rotation time,prolonged pole climbing time,a significant decrease in total movement distance within the open field(P<0.01),and a trend of shortened time spent in the central zone.The relative expression level of TH+neurons in the substantia nigra decreased significantly(P<0.05).Additionally,mRNA expression of TNF-α in the midbrain increased significantly(P<0.01),along with notable elevations in the protein expression of NF-κB(P<0.05),and Caspase-1(P<0.01).Treatment with curcumin in the fecal microbiota transplantation group of mice(model+FMTCur)showed improvements in motor abilities,evidenced by shortened pole climbing time(P<0.05),significantly prolonged rod rotation time(P<0.01),and extended time spent in the central zone(P<0.05).The relative expression level of TH+dopaminergic neurons in the substantia nigra increased significantly(P<0.05).Moreover,mRNA expression of TNF-α in the midbrain decreased significantly(P<0.01),along with notable reductions in the protein expression of NF-κB(P<0.001),p-NF-κB(P<0.01),NLRP3(P<0.05),and Caspase-1(P<0.01).Conclusion Fecal microbiota transplantation in PD model mice can induce behavioral deficits,damage TH+neurons in the substantia nigra,and trigger neuroinflammation in the brain.Subsequent curcumin treatment can ameliorate these deficits,reverse damage to TH+neurons,reduce neuroinflammatory factors,and decrease the expression of NF-κB and NLRP3 pathways.This preliminary evidence suggests that curcumin may improve Parkinsonian behavioral deficits in mice by modulating the gut microbiota.
8.Research progress in exploring cognitive processes based on pupil changes
Xiao-Ting QIAO ; Zi-Wei NI ; Bo-Zhi LIU ; Ya-Qian GUO ; Yan ZHAO ; Cai-Lian RUAN ; Ya-Yun WANG
Acta Anatomica Sinica 2025;56(3):357-363
In recent years,more and more researches has focused on the correlation between cognitive activity and physiological variables.The change of pupil is regarded as an important target in the cognitive process,and has become a hot research field.This review focuses on the three key brain regions that regulate pupil change,and reflects the neurophysiological mechanism behind pupil change by elaborating the neural pathways related to pupil change and cognitive performance.Based on recent studies on pupil change in cognitive diseases,it aims to promote the application of pupil change in the field of cognitive science in the future.
9.Design of new type of elastic gloves for burn scar rehabilitation
Ting SHEN ; Hai-yang ZHAO ; Xiao-chun JIAO ; Bao-li CHEN ; Zhi-juan WANG ; Li-na WANG ; Xiang-yang ZHAO ; Qin ZHOU
Chinese Medical Equipment Journal 2025;46(8):118-120
Objective To design a new type of elastic gloves for burn scar rehabilitation to solve the problems of conventional elastic gloves in pressure distribution,elasticity maintenance and absorption of sweat stains.Methods The new elastic gloves was made of non-woven fabric by spandex material,which was composed of external and internal parts.The main body of the external part was used as the primary structure of the gloves,which was provided with a sealing strip,a storage bag,a drawstring,etc.The internal part consisted of a bonding sheet,an elastic band,a fiber sheet,an absorbent sponge,some breathable holes,etc.Results The new elastic gloves designed could be used for the pressure therapy for the scars on the opisthenar,palm side,finger web and purlicue with scar proliferation inhibitted effectively,and the breathable hole and absorbent sponge contributed to the absorption of the sweat of the patient.Conclusion The new type of elastic gloves gains advantages in elasticity,wearing comfort and effectiveness of the pressure therapy for purlicue and finger web,and can be used for the pressure therapy to inhibit proliferative scarring after burns.[Chinese Medical Equipment Journal,2025,46(8):118-120]
10.Influence of Oatp4c1-P-gp transmembrane transport system in the kidneys of obese mice on the pharmacokinetics of digoxin
Jing-wen MEN ; Lu SUN ; Hao-ran YUE ; Zhao-yue DUAN ; Hui-nan HOU ; Ting GUI ; Jun-hong XIN ; Zhi-bo GAI
Chinese Pharmacological Bulletin 2025;41(7):1318-1324
Aim To investigate the expression of or-ganic anion transporting polypeptide 4C1(Oatp4c1)-P-glycoprotein(P-gp)in the kidneys of obese mice in-duced by high-fat diet(HFD),and its impact on the pharmacokinetic changes of digoxin.Methods C57BL/6 mice were randomly divided into the Chow group and the HFD group.Body weight and blood glu-cose were recorded weekly.After successful model es-tablishment,digoxin was intraperitoneally injected,and blood was collected at different time points.Part of the blood samples was used for LC-MS/MS detection,and the other part was used for the detection of other bio-chemical indicators.After 16 weeks,the organs were removed and weighed.HE and immunohistochemical staining was used to observe the renal pathology and the expression of Villin,a marker of proximal tubules.Western blot and qPCR were combined to detect the expression of Villin,Oatp4c1 and P-gp.Results In the HFD group,body weight and blood glucose in-creased significantly.The blood concentration of digox-in rose,the area under the curve increased,and the half-life was prolonged.The proximal tubular epithelial cells shed,and the protein expression of Villin,Oatp4c1 and P-gp decreased significantly.Conclu-sions The down-regulation of Oatp4c1-P-gp expres-sion in the kidneys of HFD mice leads to an increase in the blood concentration of digoxin and a decrease in re-nal clearance.

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