ObjectiveTo screen and identify uridine diphosphate （UDP）-rhamnose synthase （RHM） genes from Ornithogalum saundersiae and verify their UDP-rhamnose （UDP-Rha） biosynthetic functions， thereby providing reliable candidate genes for the efficient biosynthesis of neohesperidin and other rhamnoside natural products. MethodsBased on previously reported plant RHM sequences， two candidate RHM genes， OsRHM1 and OsRHM2， were identified from O. saundersiae through transcriptome screening and sequence alignment. The two genes were cloned and subjected to bioinformatics analysis and multiple sequence alignment. An in vitro RHM-Cm1，2RhaT cascade reaction was established， and the ability of OsRHM1 and OsRHM2 to catalyze the conversion of UDP-glucose （UDP-Glc） to UDP-Rha was evaluated based on neohesperidin production. Real-time fluorescence quantitative polymerase chain reaction PCR （Real-time PCR） was performed to analyze the relative expression levels of the two genes in roots， bulbs， and leaves. ResultsThe open reading frame （ORF） of OsRHM1 was 2 076 bp and encoded 692 amino acids， with a predicted protein molecular mass of 77.97 kDa and a theoretical isoelectric point of 7.25. The ORF of OsRHM2 was 2 019 bp and encoded 673 amino acids， with a predicted protein molecular mass of 75.82 kDa and a theoretical isoelectric point of 7.24. Multiple sequence alignment showed that OsRHM1 and OsRHM2 contained the typical conserved motifs GxxGxxG/A and YxxxK of the plant RHM family and shared high amino acid sequence identity with previously reported RHMs. The in vitro cascade reaction demonstrated that both OsRHM1 and OsRHM2 were capable of catalyzing the conversion of UDP-Glc to UDP-Rha. The neohesperidin production in the OsRHM2 cascade reaction system was approximately 2.58-fold that in the OsRHM1 system. Real-time PCR analysis showed that the relative expression levels of OsRHM1 and OsRHM2 were higher in roots than in bulbs and leaves. ConclusionOsRHM2 exhibited a relatively high capacity for UDP-Rha supply and may serve as a candidate enzyme resource for the subsequent biosynthesis of rhamnoside natural products and the construction of UDP-Rha supply systems.

The nod-like receptor family pyrin domain containing 3 (NLRP3) inflammasome plays a crucial role in the prognosis of subarachnoid hemorrhage (SAH). WNK1 kinase negatively regulates NLRP3 in various inflammatory conditions, but its role in early brain injury (EBI) after SAH remains unclear. In this study, we used an in vivo SAH model in rats/mice and AAV-WNK1 intraventricular injection to investigate its neuroprotective mechanisms. WNK1 expression was significantly reduced in SAH patient blood and SAH model brain tissue, correlating negatively with microglial activation. AAV-WNK1 alleviated brain edema, neuronal necrosis, behavioral deficits, and inflammation by inhibiting NLRP3 inflammasome activation. In hemin-stimulated BV-2 cells, WNK1 overexpression reduced NLRP3 activation and inflammatory cytokines. Chloride counteracted WNK1's inhibitory effects, and WNK1 suppressed P2X7R-induced NLRP3 activation. Mechanistically, WNK1 functioned via the OXSR1/STK39 pathway. These findings highlight WNK1 as a key regulator of intracellular chloride balance and neuroinflammation, presenting a potential therapeutic target for SAH treatment.
Animals ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Subarachnoid Hemorrhage/complications* ; Inflammasomes/metabolism* ; Rats ; Mice ; Neuroinflammatory Diseases/metabolism* ; WNK Lysine-Deficient Protein Kinase 1/genetics* ; Male ; Humans ; Chlorides/metabolism* ; Mice, Inbred C57BL ; Rats, Sprague-Dawley ; Brain Injuries/metabolism* ; Microglia/metabolism* ; Protein Serine-Threonine Kinases

Neuronal injury in glaucoma persists despite effective intraocular pressure (IOP) control, necessitating neuroprotective strategies for retinal ganglion cells (RGCs). In this study, we investigated the neuroprotective role of the γ-hydroxybutyrate analog HOCPCA in a glaucoma model, focusing on its effects on CaMKII signaling, oxidative stress, and neuroinflammatory responses. Retinal tissue from high IOP animal models was analyzed via proteomics. In vitro mouse retinal explants were subjected to elevated pressure and oxidative stress, followed by HOCPCA treatment. HOCPCA significantly mitigated the RGC loss induced by oxidative stress and elevated pressure, preserving neuronal function. It restored CaMKIIα and β levels, preserving RGC integrity, while also modulating oxidative stress and neuroinflammatory responses. These findings suggest that HOCPCA, through its interaction with CaMKII, holds promise as a neuroprotective therapy for glaucoma.
Animals ; Retinal Ganglion Cells/metabolism* ; Glaucoma/pathology* ; Oxidative Stress/drug effects* ; Neuroprotective Agents/pharmacology* ; Mice ; Calcium-Calmodulin-Dependent Protein Kinase Type 2/metabolism* ; Mice, Inbred C57BL ; Disease Models, Animal ; Neuroinflammatory Diseases/drug therapy* ; Neuroprotection/drug effects* ; Male ; Intraocular Pressure/drug effects*

Objective:To develop and evaluate the reliability and validity of a questionnaire for assessing long-term care needs among community-dwelling older adults.Methods:Guided by the International Classification of Functioning, Disability and Health (ICF), an initial item pool for the questionnaire was constructed based on literature review, descriptive qualitative research, and expert consultations. The draft questionnaire was formulated by transforming the established indicator system for long-term care needs into survey items. Using stratified sampling, a total of 622 older adults from community settings in Changsha were surveyed between June and August 2021. Item analysis, exploratory factor analysis (EFA), confirmatory factor analysis (CFA), and reliability testing were conducted.Results:The final questionnaire consists of three subscales (mental function, physical function, and activity and participation), comprising a total of 47 items. The item-level content validity index ( I- CVI) ranged from 0.830 to 1.000, and the scale-level content validity index ( S- CVI) was 0.915. EFA results showed that the extracted factors aligned well with the predefined dimensions after adjustment, with factor loadings exceeding 0.600 and cumulative variance contributions exceeding 70.000% for all subscales. CFA results indicated good model fit across all subscales and the overall questionnaire, confirming stable structural validity. The Cronbach's α coefficients ranged from 0.877 to 0.970, and test-retest reliability coefficients ranged from 0.901 to 0.957. Conclusions:The long-term care needs assessment questionnaire for community-dwelling older adults demonstrates strong reliability and validity, and can serve as a reliable tool for evaluating long-term care needs in this population.

Objective To observe the efficacy,safety,and compliance of lamotrigine combined with magnesium valproate in treating children with depressive episodes of bipolar disorder,and to explore the effects on thyroid hormone levels,brain-derived neurotrophic factor(BDNF),C-reactive protein(CRP),interleukin-1(IL-1),interleukin-10(IL-10),tumor necrosis factor-α(TNF-α)and plasma concentration of valproate.Methods The children with bipolar disorder diagnosed from January 2023 to February 2025 were selected,and divided into the observation group and control group.The control group was treated with magnesium valproate tablets,and the observation group was added lamotrigine in addition to the treatment given to the control group.Both groups were treated continuously for 8 weeks.The clinical efficacy,the Hamilton Depression Scale-24(HAMD-24)score,Clinical Global Impression(CGI)assessment,thyroid hormone levels,BDNF,CRP,IL-1,IL-10 and TNF-α,daily average dose of magnesium valproate(D),blood concentration of valproate(C),C/D ratio,mean dosing interval(h),incidence of adverse reactions,and medication adherence and satisfaction scores in both groups was observed.Results A total of 100 children were included,50 in each group.After treatment,the total effective rate of the observation group was 98.00％which was significantly higher than that of the control group(84.00％)(P＜0.05).The serum free triiodothyronine(FT3),free thyroxine(FT4),BDNF and IL-10 in both groups increased compared to the previous(P＜0.05),while HAMD-24 score,CGI score,thyroid-stimulating hormone(TSH),CRP,IL-1 and TNF-α decreased(P＜0.05),and all indicators in the observation group were better than those in the control group(P＜0.05).Both groups had no serious or new adverse drug reactions,and the incidence of total adverse reactions,the difference in the incidence of total adverse reactions was not statistically significant(P＞0.05).There was no statistically significant difference in valproic acid blood concentrations between the two groups of children(P＞0.05).The medication compliance score and satisfaction score of the observation group were significantly higher than those of the control group(P＜0.05).Conclusion Combination of lamotrigine with magnesium valproate in children with depressive episodes of bipolar disorder improves treatment effective and clinical symptoms,promotes a rise in thyroid hormone and BDNF as well as improves inflammatory factors and increases medication adherence and satisfaction in children with a better safety profile.

Objective To observe the efficacy,safety,and compliance of lamotrigine combined with magnesium valproate in treating children with depressive episodes of bipolar disorder,and to explore the effects on thyroid hormone levels,brain-derived neurotrophic factor(BDNF),C-reactive protein(CRP),interleukin-1(IL-1),interleukin-10(IL-10),tumor necrosis factor-α(TNF-α)and plasma concentration of valproate.Methods The children with bipolar disorder diagnosed from January 2023 to February 2025 were selected,and divided into the observation group and control group.The control group was treated with magnesium valproate tablets,and the observation group was added lamotrigine in addition to the treatment given to the control group.Both groups were treated continuously for 8 weeks.The clinical efficacy,the Hamilton Depression Scale-24(HAMD-24)score,Clinical Global Impression(CGI)assessment,thyroid hormone levels,BDNF,CRP,IL-1,IL-10 and TNF-α,daily average dose of magnesium valproate(D),blood concentration of valproate(C),C/D ratio,mean dosing interval(h),incidence of adverse reactions,and medication adherence and satisfaction scores in both groups was observed.Results A total of 100 children were included,50 in each group.After treatment,the total effective rate of the observation group was 98.00％which was significantly higher than that of the control group(84.00％)(P＜0.05).The serum free triiodothyronine(FT3),free thyroxine(FT4),BDNF and IL-10 in both groups increased compared to the previous(P＜0.05),while HAMD-24 score,CGI score,thyroid-stimulating hormone(TSH),CRP,IL-1 and TNF-α decreased(P＜0.05),and all indicators in the observation group were better than those in the control group(P＜0.05).Both groups had no serious or new adverse drug reactions,and the incidence of total adverse reactions,the difference in the incidence of total adverse reactions was not statistically significant(P＞0.05).There was no statistically significant difference in valproic acid blood concentrations between the two groups of children(P＞0.05).The medication compliance score and satisfaction score of the observation group were significantly higher than those of the control group(P＜0.05).Conclusion Combination of lamotrigine with magnesium valproate in children with depressive episodes of bipolar disorder improves treatment effective and clinical symptoms,promotes a rise in thyroid hormone and BDNF as well as improves inflammatory factors and increases medication adherence and satisfaction in children with a better safety profile.

Objective:To develop and evaluate the reliability and validity of a questionnaire for assessing long-term care needs among community-dwelling older adults.Methods:Guided by the International Classification of Functioning, Disability and Health (ICF), an initial item pool for the questionnaire was constructed based on literature review, descriptive qualitative research, and expert consultations. The draft questionnaire was formulated by transforming the established indicator system for long-term care needs into survey items. Using stratified sampling, a total of 622 older adults from community settings in Changsha were surveyed between June and August 2021. Item analysis, exploratory factor analysis (EFA), confirmatory factor analysis (CFA), and reliability testing were conducted.Results:The final questionnaire consists of three subscales (mental function, physical function, and activity and participation), comprising a total of 47 items. The item-level content validity index ( I- CVI) ranged from 0.830 to 1.000, and the scale-level content validity index ( S- CVI) was 0.915. EFA results showed that the extracted factors aligned well with the predefined dimensions after adjustment, with factor loadings exceeding 0.600 and cumulative variance contributions exceeding 70.000% for all subscales. CFA results indicated good model fit across all subscales and the overall questionnaire, confirming stable structural validity. The Cronbach's α coefficients ranged from 0.877 to 0.970, and test-retest reliability coefficients ranged from 0.901 to 0.957. Conclusions:The long-term care needs assessment questionnaire for community-dwelling older adults demonstrates strong reliability and validity, and can serve as a reliable tool for evaluating long-term care needs in this population.

Objective:Laboratory evaluation of the relationship between cerebrospinal fluid and plasma indicators and intrathecal immunoglobulin G(IgG) synthesis in patients with neurological diseases, and establishment of a new diagnostic method for intrathecal IgG synthesis.Methods:This study retrospectively analyzed the content of IgG in cerebrospinal fluid samples and blood albumin in blood samples, and other test results of 410 patients with neurological diseases who visited Beijing Tiantan Hospital from 2019 to 2022. According to the results of oligoclonal bands in cerebrospinal fluid, patients were divided into intrathecal IgG synthesis group and non-intrathecal IgG synthesis group. The Mann Whitney U test was used for inter group comparison, and a bilateral test with P<0.05 indicates a statistically significant difference. Include indicators with differences between groups in logistic regression analysis, construct a predictive model, and compare it with the established quantitative formula IgG index. Results:There were significant differences in 10 indicators, including cerebrospinal fluid leukocyte count and 24-hour intrathecal IgG synthesis rate, between the intrathecal IgG synthesis group and the non-intrathecal IgG synthesis group, with P<0.05. The area under the curve (AUC) of intrathecal IgG synthesis was higher than the IgG index (AUC=0.920, 0.809, Z=31.178, P<0.001), the sensitivity was higher than the IgG index (0.825, 0.618), and the specificity was lower than the IgG index (0.876, 0.908). Conclusion:The combination of 10 indicators such as cerebrospinal fluid white blood cell count and 24-hour intrathecal IgG synthesis rate can improve the diagnostic efficacy and sensitivity of intrathecal IgG synthesis.

Objective:To construct an evaluation index system for the long-term care needs of community-dwelling elderly.Methods:Based on the International Classification of Functioning, Disability and Health, a preliminary evaluation index system was formed using literature research and descriptive research. The evaluation index system was revised and improved through the expert consultation.Results:Two rounds of consultation were conducted on 20 experts. The effective response rates of the questionnaire for two rounds of expert consultation were 90.0% (18/20) and 94.4% (17/18), respectively. The expert opinion submission rates were 94.4% (17/18) and 29.4% (5/17), and the expert authority coefficients were 0.94 and 0.95, respectively. After two rounds of expert consultation, the coefficient of variation of each indicator was less than 0.15, and the Kendall harmony coefficients of the overall indicator and all levels of indicators were 0.277 to 0.441 (all P<0.01). The final evaluation index system consisted of three first-level indicators, 12 second-level indicators, and 49 third-level indicators. Conclusions:The construction process of the evaluation index system established is rigorous, the content is comprehensive, and the research methods and results are scientific and reliable. It can provide a basis for further developing long-term care needs assessment tools for community-dwelling elderly and establishing grading standards for long-term care needs assessment.

Circadian rhythm disorders are closely related to metabolic diseases， which can cause non-alcoholic fatty liver disease （NAFLD） by directly acting on the liver or indirectly affecting the liver through the liver-brain axis and intestinal flora. The rhythms of yin and yang， ying （营） and wei （卫）， twelve hours and four seasons in traditional Chinese medicine （TCM） chronomedicine are similar to the connotations of modern biological rhythms. From the perspective of chronomedicine of TCM， the incidence of NAFLD is closely related to the abnormality of the daily rhythm of the waxing and waning of yin and yang， the daily rhythm of the circulation of ying and wei， the rhythm of twelve hours and four seasons. Through analyzing the rhythms related to the occurrence， development and prognosis of NAFLD， it is helpful to enhance the understanding of NAFLD in relation to time， so as to better guide the clinical diagnosis and treatment.