ObjectiveTo investigate the effects of Chaihu Jia Longgu Mulitang（CJLM） on depression-like behaviors and neuroinflammation in rats subjected to social isolation combined with chronic unpredictable mild stress（CUMS）， and to explore the potential underlying mechanisms. MethodsSixty male SD rats were randomly divided into a normal group， a model group， and low-， medium-， and high-dose CJLM groups（2.89， 5.78， 11.56 g·kg^-1）， as well as a fluoxetine group（10 mg·kg^-1）. Except for the normal group， all other groups were subjected to social isolation combined with CUMS for 63 d. During the first 35 d， depression models were established only， and from day 36 onward， modeling and drug administration were conducted simultaneously for a total intervention period of 28 d. Depression-like behaviors were evaluated using the sucrose preference test， open-field test， and forced swimming test. Hematoxylin-eosin（HE） staining was performed to observe hippocampal histomorphology. Immunohistochemistry（IHC） was used to detect the expression levels of ionized calcium-binding adapter molecule 1（Iba1） and gasdermin D（GSDMD） proteins in the hippocampus. Western blot analysis was employed to determine the protein expression levels of adenosine 5′-monophosphate（AMP）-activated protein kinase（AMPK） and phosphorylated（p）-AMPK， silent information regulator 1（SIRT1）， nuclear factor-κB（NF-κB） and p-NF-κB， NOD-like receptor protein 3（NLRP3）， and Caspase-1 in the hippocampus. Real-time quantitative polymerase chain reaction（Real-time PCR） was used to detect the mRNA expression levels of tumor necrosis factor-α（TNF-α）， interleukin（IL）-6， and IL-1β in the hippocampus. ResultsCompared with the normal group， the model group showed a decreased sucrose preference rate（P<0.01）， reduced total movement distance（P<0.01）， prolonged immobility time（P<0.01）， and decreased central zone residence time（P<0.01） in the open-field test， and increased immobility time in the forced swimming test（P<0.01）. Hippocampal neuronal structure was damaged. The contents of Iba1 and GSDMD in the hippocampus were significantly increased（P<0.01）. The protein expression levels of p-AMPK and SIRT1 in the hippocampus were significantly decreased（P<0.01）， whereas the protein expression levels of p-NF-κB， NLRP3， and Caspase-1 were significantly increased（P<0.01）. The mRNA expression levels of IL-1β， IL-6， and TNF-α in the hippocampus were significantly upregulated（P<0.01）. Compared with the model group， the low-， medium-， and high-dose CJLM groups and the fluoxetine group all were able to reverse depression-like behavioral changes， as evidenced by increased sucrose preference rate， increased total movement distance with shortened immobility time in the open-field test， prolonged central zone residence time， and reduced immobility time in the forced swimming test（P<0.05， P<0.01）. Meanwhile， hippocampal neuronal structural damage was alleviated. In the hippocampus， the expression levels of Iba1 and GSDMD were downregulated， the expression levels of p-AMPK and SIRT1 were upregulated， and the abnormal elevations of p-NF-κB， NLRP3， Caspase-1， as well as IL-1β， IL-6， and TNF-α mRNA were suppressed（P<0.05， P<0.01）. ConclusionCJLM can ameliorate depression-like behaviors in rats subjected to social isolation combined with CUMS and attenuate hippocampal neuroinflammation and pyroptosis， suggesting that its effects may be associated with the regulation of AMPK/SIRT1/NF-κB/NLRP3 signaling pathway.

Objective:To establish a method for determination of fluoride in water using a fully automated biochemical analyzer (abbreviated as this method).Methods:Based on the parameters of the fully automatic biochemical analyzer, appropriate reagent volumes and reaction time were optimized to determine fluoride ions in water within the standard curve range (0.0 - 2.0 mg/L). The method was validated through evaluation of linear range, detection limit, precision, accuracy, and comparative analysis with the manual measurement results of fluoride reagent spectrophotometric method specified in the "Standard Examination Methods for Drinking Water - Part 5: Inorganic Nonmetallic indices" (GB/T 5750.5-2023).Results:Within the range of 0.0 - 2.0 mg/L fluoride mass concentration, the absolute values of the correlation coefficients were > 0.999 0 and the detection limit of this method was 0.082 mg/L. The relative standard deviation for the determination of fluoride level in water samples with low, medium and high fluoride mass concentrations was 4.03%, 2.16% and 1.68%, respectively. The spiked recovery rates were 98.1%, 99.5%, and 100.2%, respectively. There was no statistically significant difference between the results obtained by this method and manual measurement ( t = 1.07, P = 0.295). Conclusion:This method exhibits high sensitivity, accuracy, and efficiency in detecting fluoride, enabling rapid sample detection through instrumental automation instead of manual operation.

Objective:To establish a method for determination of fluoride in water using a fully automated biochemical analyzer (abbreviated as this method).Methods:Based on the parameters of the fully automatic biochemical analyzer, appropriate reagent volumes and reaction time were optimized to determine fluoride ions in water within the standard curve range (0.0 - 2.0 mg/L). The method was validated through evaluation of linear range, detection limit, precision, accuracy, and comparative analysis with the manual measurement results of fluoride reagent spectrophotometric method specified in the "Standard Examination Methods for Drinking Water - Part 5: Inorganic Nonmetallic indices" (GB/T 5750.5-2023).Results:Within the range of 0.0 - 2.0 mg/L fluoride mass concentration, the absolute values of the correlation coefficients were > 0.999 0 and the detection limit of this method was 0.082 mg/L. The relative standard deviation for the determination of fluoride level in water samples with low, medium and high fluoride mass concentrations was 4.03%, 2.16% and 1.68%, respectively. The spiked recovery rates were 98.1%, 99.5%, and 100.2%, respectively. There was no statistically significant difference between the results obtained by this method and manual measurement ( t = 1.07, P = 0.295). Conclusion:This method exhibits high sensitivity, accuracy, and efficiency in detecting fluoride, enabling rapid sample detection through instrumental automation instead of manual operation.

OBJECTIVE To analyze the chemical constituents of Heihuang Chizhu Granules and the blood composition of rats af-ter administration by UPLC-Q-TOF-MS/MS.METHODS A Waters ACQUITY UPLC HSS T3 column(3 mm×100 mm,1.8 μm)was eluted with acetonitrile-0.1%formic acid as mobile phase,and the data were collected in electrospray ion source positive and neg-ative ion mode and then identified with the reference retention time,precise molecular weight,secondary fragment ions,and references to relevant literature.RESULTS A total of 104 chemical components were identified from Heihuang Chizhu Granules,including 26 flavonoids,24 organic acids,14 triterpenoids,8 terpenoids,7 phenylpropanoids,11 monoglycosides,and 14 other components(phe-nols,alkaloids,etc.).On this basis,39 blood-entering components were identified in the plasma of rats administered via gavage,in-cluding 28 prototypes and 11 metabolites.CONCLUSION The chemical constituents of Heihuang Chizhu Granules and the compo-nents entering the blood of rats are analyzed and identified for the first time in this study,and the results provide a scientific basis for the basic research of Heihuang Chizhu Granules and the establishment of process quality control standards.

Objective To investigate the impact of mitophagy,mediated by the long non-coding RNA SNHG16(LncRNA SNHG16)on diabetes-associated cognitive impairment(DCI).Methods 29 male C57BL/J mice were randomly divided into normal control(NC)group,DCI group and DCI+mitochondrial autophagy inhibitor(DCI+Mdivi-1)group.Morris water maze and new object recognition test were used to detect the cognitive function of mice,qRT-CPR was used to detect the expression of LncRNA SNHG16 and mitochondrial autophagy marker mRNA.Western blot were used to detect the expression of related protein.The mouse hippocampal neurons HT22 were divided into control(Con)group,high glucose(HG)group,HG+SNHG16 silencing(HG+sh-SNHG16)group and HG+no-load control(HG+sh-NC)group.CCK8 method and lactate dehydrogenase(LDH)method were used to detect neuronal damage.JC-1 method was used to detect mitochondrial membrane potential.Results Compared with NC group,the expression of LncRNA SNHG16 and the expression of autophagy-related gene 5,PTEN-induced putative kinase 1(PINK1),Parkin and microtubule associated protein light chain 3(LC3)Ⅱ/Ⅰ increased(P<0.05),while the expression of mitochondrial autophagy-related proteins P62 and mitochondrial outer membrane transposase 20(TOMM20)decreased in T2DM group.Compared with DCI group,the cognitive dysfunction of mice improved,and the expression level of LncRNA SNHG16 decreased in the DCI+Mdivi-1 group(P<0.05).The expressions of LncRNA SNHG16,LC3 Ⅱ/Ⅰ,PINK1 and Parkin were higher in HG group than in Con group(P<0.05),while the cell survival rate and TOMM20 protein expression were lower in HG group than in Con group(P<0.05).Silence of LncRNA SNHG16 can restore the activity of HT22 cells and mitochondrial membrane potential,and reduce the level of mitochondrial autophagy under HG condition.Conclusions The expression level of LncRNA SNHG16 was up-regulated in the hippocampus brain region of mice with diabetic cognitive dysfunction,and mitophagy was overactivated.Silencing of LncRNA SNHG16 inhibits mitophagy in hippocampal neurons and alleviates HG induced hippocampal neuronal damage.

[Objective]To explore the effects of astragalin(AST)on autophagy and apoptosis of astrocytes in the L4-5 dorsal horn of the spinal cord in mice with inflammatory pain induced by complete Freund's adjuvant(CFA).[Methods]Twenty-four male C57BL/6 mice,aged six months,were randomly assigned to four groups:control group,saline group,CFA model group,and CFA+AST group,six mice in each group.The inflammatory pain model was established by injection of 10 μL CFA into the right lateral malleolus fossa.The saline group were injected with an equal amount of normal saline at the same site.The inflammatory pain mice in CFA+AST group were further treated with AST(60 mg/kg)intraperitoneally once a day for 21 consecutive days.Multiplex immunofluorescence staining was used to detect the coexpression of autophagy-related factors including ATG 12 and Beclin-1,apoptosis-related factors including Cleaved-Caspase3 and Caspase9,and the astrocyte marker such as GFAP in the L4-5 spinal dorsal horn of the mice in each group.Western blot was used to examine the protein expression levels of autophagy-related proteins(ATG12,Beclin-1)and apoptosis-related proteins(Caspase 3,Caspase 9)in the L4-5 spinal dorsal horn of mice.[Results]Immunofluorescent staining showed that in the L4-5 dorsal horn of the spinal cord,the fluorescence intensity of ATG12(P<0.000 1)and Beclin-1(P<0.000 1)was significantly increased,while that of Cleaved-Caspase 3(P<0.001)and Caspase 9(P<0.000 1)was decreased in the CFA+AST group when compared to the CFA model group.Furthermore,AST could inhibit the activation of astrocytes.Western blot further confirmed that AST significantly upregulated the expression of ATG12(P<0.000 1)and Beclin-1(P<0.000 1)in the L4-5 spinal cord of CFA mice,and downregulated the expression of Caspase 3(P<0.01)and Caspase 9(P<0.001).[Conclusions]AST promotes autophagy of astrocytes and inhibits their apoptosis in the L4-5 spinal dorsal horn of CFA mice.

Objective To analyze the research hotspots,frontiers,and trends in clinical randomized controlled trials(RCTs)on acupuncture-moxibustion for post-stroke shoulder-hand syndrome(SHS)over the past decade using bibliometric and knowledge mapping methods.Methods The clinical RCTs on acupuncture-moxibustion for post-stroke SHS from five Chinese and English databases were retrieved,including CNKI,Wanfang,VIP,PubMed,and Web of Science(WOS).CiteSpace and VOSviewer were employed for analysis,and visualized knowledge maps were generated.Results A total of 1 226 Chinese and 20 English articles were included,with both reaching publication peaks in 2021.The top three institutions in terms of output were Huguosi Traditional Chinese Medicine Hospital Affiliated to Beijing University of Chinese Medicine,The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine,and Heilongjiang University of Chinese Medicine.The most prolific author was Song Fengjun,while Pei Jian had the highest centrality.The five most frequently used keywords were shoulder-hand syndrome,stroke,acupuncture-moxibustion,rehabilitation training,and joint mobility.Emerging research topics included activities of daily living,limb function,and range of motion.Conclusion Clinical research on acupuncture-moxibustion for post-stroke SHS holds significant developmental potential.Strengthening international collaboration to explore therapeutic mechanisms is recommended,and researchers should continue optimizing treatment protocols to enhance the quality of clinical evidence.

Objective:To analyze the results of national external quality assessment (EQA) for transfusion compatibility test in 2023, and provide reference for quality management of clinical transfusion compatibility testing.Methods:The EQA of clinical transfusion compatibility testing by NCCL was performed 3 times in 2023 among included laboratories. The panel consisting of 22 samples was distributed to 4 186 laboratories across 31 provinces (Including 2 961 tertiary hospital laboratories, 1 085 secondary hospital laboratories, 23 primary hospital laboratories, 106 blood station laboratories and 11 independent clinical laboratories). Each panel contains 11 red blood cell and 11 plasma samples per 1.5 ml/tube. Each participant laboratory of the EQA program was required to carry out the detection and return results in expected time. Statistical analysis and evaluation on the reported results were conducted by NCCL from the aspects of regional distribution, laboratory grading, testing methodology, reagent and testing system usage.Results:The qualification rates of EQA for five items including ABO positive typing, ABO reverse typing, RhD blood type, antibody screening, and cross matching were 96.68%, 95.10%, 96.46%, 95.32%, and 91.04%, respectively. The EQA qualification rate of tertiary hospital laboratories was 87.77% (2 599/2 961), which was significantly higher than the 77.79% (844/1 085) of secondary hospital laboratories. There were significant differences in the qualification rate of participating laboratories among different regions. The utilization rates of micro column agglutination method in ABO positive typing, ABO reverse typing, RhD blood type, antibody screening, and cross matching were 80.81% (10 080/12 474), 75.06% (9 337/12 440), 81.38% (10 118/12 433), 89.59% (11 104/12 394) and 76.25% (9 495/12 453), respectively. The qualification rate of micro column agglutination method was significantly higher than that of saline slide method in ABO positive typing detection ( P<0.05). The qualification rate of micro column agglutination method was significantly higher than that of the polyamine method and anti-human globulin test tube method in antibody screening ( P<0.05). There were statistically significant differences in qualification rate of 7 reagents in ABO reverse typing, antibody screening and cross matching ( P<0.05). There was no statistically significant difference in the qualification rate between the two detection systems for other reagents, except for the ABO reverse typing where the qualification rate of reagent 1 in a single system was higher than that in a mixed system ( P<0.05). Conclusion:The testing capabilities of clinical laboratories in different regions and different type varied significantly in China. Micro column agglutination method was the most popular selection in transfusion compatibility testing. The regents used in these laboratories showed good performance. However, the detection efficiency of some reagents still need to be improved. EQA could be used to evaluate, monitor, and improve the quality of testing.

Objective:To establish and optimize an autoverification system for thyroid function test reports of 5 items using historical test data.Methods:Based on the docoment' Autoverification of Clinical Laboratory Quantitative Test Results′, CLSI AUTO 10-A and AUTO 15 guidelines, an autoverification system for thyroid function test reports of 5 items was established combining with manual verification experience. A total of 193 860 thyroid function test reports of 5 items in 2021 were collected for the assessment of the original system. Totally 210 097 thyroid function test reports of 5 items in 2022 and 299 198 reports in 2023 were collected for the optimization of the autoverification system. There were 160 666 thyroid function test reports of 5 items from the first half of 2024 for the manual and autoverification comparison after optimization.Results:The pass rate of the autoverification system based on original thyroid function report in 2021 was 69.56%(134 849/193 860). The optimized system utilizing historical data from 2022 and 2023 covered 21 pattern rules and established verification for different patterns including range rules, delta check rules, and review rules. Taking manual verification as the standard for the data from the first half of 2024, the sensitivity and specificity of the optimized system were 100% (499/499) and 81.57% (130 646/160 167), respectively, with a false-negative rate of 0. The concordance rate between autoverification and manual verification was 81.63% (131 145/160 666), and the pass rate was 81.32% (130 646/160 666).Conclusion:Establishing and optimizing the autoverification system for thyroid function tests of 5 items using historical test data, and formulating verification rules for different patterns can be applied to clinical practise, which not only ensures the accuracy of test reports but also improves work efficiency, allowing continuously optimized and perfected of the system.

Existing emotion recognition research is typically limited to static laboratory settings and has not fully handle the changes in emotional states in dynamic scenarios. To address this problem, this paper proposes a method for dynamic continuous emotion recognition based on electroencephalography (EEG) and eye movement signals. Firstly, an experimental paradigm was designed to cover six dynamic emotion transition scenarios including happy to calm, calm to happy, sad to calm, calm to sad, nervous to calm, and calm to nervous. EEG and eye movement data were collected simultaneously from 20 subjects to fill the gap in current multimodal dynamic continuous emotion datasets. In the valence-arousal two-dimensional space, emotion ratings for stimulus videos were performed every five seconds on a scale of 1 to 9, and dynamic continuous emotion labels were normalized. Subsequently, frequency band features were extracted from the preprocessed EEG and eye movement data. A cascade feature fusion approach was used to effectively combine EEG and eye movement features, generating an information-rich multimodal feature vector. This feature vector was input into four regression models including support vector regression with radial basis function kernel, decision tree, random forest, and K-nearest neighbors, to develop the dynamic continuous emotion recognition model. The results showed that the proposed method achieved the lowest mean square error for valence and arousal across the six dynamic continuous emotions. This approach can accurately recognize various emotion transitions in dynamic situations, offering higher accuracy and robustness compared to using either EEG or eye movement signals alone, making it well-suited for practical applications.
Humans ; Electroencephalography/methods* ; Emotions/physiology* ; Eye Movements/physiology* ; Signal Processing, Computer-Assisted ; Support Vector Machine ; Algorithms