ObjectiveTo investigate the mechanism by which 2，3，5，4'-tetrahydroxyldiphenylethylene-2-O-glucoside （THSG） mitigates cerebral ischemia/reperfusion （CI/R） injury by regulating mitochondrial calcium overload and promoting mitophagy. MethodsSixty male SD rats were randomized into sham， model， SAS （40 mg·kg^-1）， and low-， medium- and high-dose （10， 20， 40 mg·kg^-1， respectively） THSG groups， with 10 rats in each group. The middle cerebral artery occlusion/reperfusion （MCAO/R） model was established by the modified Longa suture method. An oxygen-glucose deprivation/reoxygenation （OGD/R） model was constructed in PC12 cells. Neurological deficits were assessed via Zea Longa scoring， and cerebral infarct volume was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Structural and functional changes of cortical neurons in MCAO/R rats were assessed by hematoxylin-eosin and Nissl staining. PC12 cell viability was detected by cell counting kit-8 （CCK-8） assay， and mitochondrial calcium levels were quantified by Rhod-2 AM. Immunofluorescence was used to detect co-localization of PTEN-induced kinase 1 （PINK1） and leucine zipper/EF-hand-containing transmembrane protein 1 （LETM1） in neurons. Transmission electron microscopy （TEM） was employed to observe mitochondrial morphology in neurons. Western blot was employed to analyze the expression of translocase of outer mitochondrial membrane 20 （TOMM20）， autophagy-associated protein p62， microtubule-associated protein light chain 3 （LC3）， cysteinyl aspartate-specific proteinase-9 （Caspase-9）， B-cell lymphoma 2-associated protein X （Bax）， and cytochrome C （Cyt C）. ResultsCompared with the sham group， the model group exhibited increased infarct volume （P<0.01） and neurological deficit scores （P<0.01）， neuronal structure was disrupted with reduced Nissl bodies. （P<0.01）， mitochondrial swelling/fragmentation， decreased PINK1/LETM1 co-localization （P<0.01）， upregulated protein levels of LC3Ⅱ/LC3Ⅰ， TOMM20， Caspase-9， Bax， and Cyt C （P<0.01）， downregulated protein level of p62 （P<0.05）， weakened PC12 viability （P<0.01）， and elevated mitochondrial calcium level （P<0.01）. Compared with the model group， THSG and SAS groups showed reduced infarct volumes （P<0.05，P<0.01） and neurological deficit scores （P<0.05，P<0.01）， mitigated mitochondrial damage， and increased PINK1/LETM1 co-localization （P<0.01）. Medium/high-dose THSG and SAS alleviated the neurological damage， increased Nissl bodies （P<0.05，P<0.01）， downregulated the protein levels of p62， TOMM20， Caspase-9， Bax， and Cyt C （P<0.05，P<0.01）， and elevated the LC3Ⅱ/LC3Ⅰ level （P<0.05，P<0.01）. High-dose THSG enhanced PC12 cell viability （P<0.01）， increased PINK1/LETM1 co-localization （P<0.01）， and reduced mitochondrial calcium （P<0.01）. ConclusionTHSG may exert the neuroprotective effect on CI/R injury by activating the PINK1-LETM1 signaling pathway， reducing the mitochondrial calcium overload， and promoting mitophagy.

ObjectiveTo investigate the mechanism by which 2，3，5，4'-tetrahydroxyldiphenylethylene-2-O-glucoside （THSG） mitigates cerebral ischemia/reperfusion （CI/R） injury by regulating mitochondrial calcium overload and promoting mitophagy. MethodsSixty male SD rats were randomized into sham， model， SAS （40 mg·kg^-1）， and low-， medium- and high-dose （10， 20， 40 mg·kg^-1， respectively） THSG groups， with 10 rats in each group. The middle cerebral artery occlusion/reperfusion （MCAO/R） model was established by the modified Longa suture method. An oxygen-glucose deprivation/reoxygenation （OGD/R） model was constructed in PC12 cells. Neurological deficits were assessed via Zea Longa scoring， and cerebral infarct volume was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Structural and functional changes of cortical neurons in MCAO/R rats were assessed by hematoxylin-eosin and Nissl staining. PC12 cell viability was detected by cell counting kit-8 （CCK-8） assay， and mitochondrial calcium levels were quantified by Rhod-2 AM. Immunofluorescence was used to detect co-localization of PTEN-induced kinase 1 （PINK1） and leucine zipper/EF-hand-containing transmembrane protein 1 （LETM1） in neurons. Transmission electron microscopy （TEM） was employed to observe mitochondrial morphology in neurons. Western blot was employed to analyze the expression of translocase of outer mitochondrial membrane 20 （TOMM20）， autophagy-associated protein p62， microtubule-associated protein light chain 3 （LC3）， cysteinyl aspartate-specific proteinase-9 （Caspase-9）， B-cell lymphoma 2-associated protein X （Bax）， and cytochrome C （Cyt C）. ResultsCompared with the sham group， the model group exhibited increased infarct volume （P<0.01） and neurological deficit scores （P<0.01）， neuronal structure was disrupted with reduced Nissl bodies. （P<0.01）， mitochondrial swelling/fragmentation， decreased PINK1/LETM1 co-localization （P<0.01）， upregulated protein levels of LC3Ⅱ/LC3Ⅰ， TOMM20， Caspase-9， Bax， and Cyt C （P<0.01）， downregulated protein level of p62 （P<0.05）， weakened PC12 viability （P<0.01）， and elevated mitochondrial calcium level （P<0.01）. Compared with the model group， THSG and SAS groups showed reduced infarct volumes （P<0.05，P<0.01） and neurological deficit scores （P<0.05，P<0.01）， mitigated mitochondrial damage， and increased PINK1/LETM1 co-localization （P<0.01）. Medium/high-dose THSG and SAS alleviated the neurological damage， increased Nissl bodies （P<0.05，P<0.01）， downregulated the protein levels of p62， TOMM20， Caspase-9， Bax， and Cyt C （P<0.05，P<0.01）， and elevated the LC3Ⅱ/LC3Ⅰ level （P<0.05，P<0.01）. High-dose THSG enhanced PC12 cell viability （P<0.01）， increased PINK1/LETM1 co-localization （P<0.01）， and reduced mitochondrial calcium （P<0.01）. ConclusionTHSG may exert the neuroprotective effect on CI/R injury by activating the PINK1-LETM1 signaling pathway， reducing the mitochondrial calcium overload， and promoting mitophagy.

Anxiety disorder is a common emotional disorder,with its pathological mechanisms involving the imbalance of multiple neurotransmitter systems,especially the dysfunction of the gamma-aminobutyric acid(GABA)system.This article explored the TCM understanding of anxiety disorder,and summarized and organized the research on the regulation of GABA system in the treatment of anxiety disorders through TCM monomers and formulas from the aspects of terpenes,flavonoids,phenolic acids,lignans,cinnamic acids,saponins,etc.At present,the research mainly involved chrysin,baicalin,ginsenoside,Baihe Dihuang Decoction,Bentun Decoction,Suanzaoren Decoction,Chaihu Jia Longgu Muli Decoction,etc.The mechanism involved regulating the inflammatory response of the body,alleviating oxidative stress damage,restoring the hyperactive HPA axis and other action pathways,which could provide a reference for the related research of TCM anti anxiety treatment and the development of new drugs.

Objective:To construct a hierarchical training course system for neonatal nurses based on core competencies, to provide a reference for meeting the training needs of neonatal nurses under the new situation.Methods:Through literature review, questionnaire survey on training needs, and focus group interviews, a preliminary hierarchical training curriculum system for neonatal nurses was developed. Two rounds of Delphi correspondence were conducted with 19 domestic experts to finalize the system.Results:The effective questionnaire recovery rates of the two rounds of expert consultation were 95.00% and 100.00%, and the expert authority coefficient was 0.916, the Kendall harmony coefficient of the first round of expert opinions was 0.351 ( P<0.001), and the Kendall harmony coefficient of the second round of expert opinions was 0.463 ( P<0.001). The hierarchical training course structure and course training content are formed, including N0: 3 first-level items, 9 second-level items, 80 third-level items, N1: 3 first-level items, 9 second-level items, 91 third-level items, N2: 3 first-level items, 9 second-level items, 86 third-level items, N3: 3 first-level items, 10 second-level items, 81 third-level items, N4: 3 first-level items, 10 second-level items, 76 third-level items. Conclusions:The hierarchical training course system for neonatal nurses based on the core competence of nurses is scientific and practical, which can provide a reference for the hierarchical training of neonatal nurses.

Objective:To investigate the numbers of peripheral blood CD4 +T cell subpopulations in patients with elderly-onset rheumatoid arthritis (EORA) and its clinical significance. Methods:A total of 188 patients with newly diagnosed RA in the department of rheumatology and immunology of the Second Hospital of Shanxi Medical University from January 2020 to December 2023 were collected, including 48 cases of EORA (age of onset: ≥60 years old), 140 cases of young-onset rheumatoid arthritis (YORA) (18 years old ≤ age of onset < 60 years old). Meanwhile, 151 healthy controls (HC) were collected, of which 31 persons aged 60-85 years were included as HC group 1 (HC 1) and 120 individuals aged 18-59 years were included as HC group 2 (HC 2). Peripheral blood CD4 +T lymphocyte subsets of these participants were assessed by flow cytometry. Differences between groups were analyzed using independent-samples t test, Mann-Whitney U test or χ2 test. Results:Compared with healthy individuals, the absolute counts and percentages of peripheral blood Treg cells in patients with EORA were significantly decreased [absolute counts: 32.65 (23.04, 47.73) cells/μl vs. 23.03 (15.28, 32.12) cells/μl, Z=-3.35, P=0.001; percentages: 5.12%(4.13%, 6.16%) vs. 3.72% (2.79%, 4.82%), Z=-4.10, P<0.001], while the Th17/Treg cell ratio was increased [0.16 (0.12, 0.29) vs. 0.26 (0.18, 0.46), Z=-2.94, P=0.003], the differences are all significant. There was a tendency with higher absolute counts and percentages of Treg [absolute counts: 23.03 (15.28, 32.12) cells/μl vs. 20.97 (14.01, 30.64) cells/μl, Z=-0.58, P=0.561; percentages: 3.72%(2.79%, 4.82%) vs. 3.38% (2.39%, 4.71%), Z=-1.06, P=0.287] and lower Th17/Treg ratios [0.26 (0.18, 0.46) vs. 0.27 (0.19, 0.46), Z=-0.32, P=0.751] in EORA when compared to patients with YORA, but no significant differences were observed. Conclusion:Patients with EORA also have the reduced numbers of peripheral blood Treg cells and immune imbalance of Th17/Treg, suggesting that immune imbalance or dysfunction caused by defects in Treg cell counts and/or function contributes to the development of EORA, and that targeting Treg cells may be a promising therapeutic strategy for EORA.

Objective:To construct a hierarchical training course system for neonatal nurses based on core competencies, to provide a reference for meeting the training needs of neonatal nurses under the new situation.Methods:Through literature review, questionnaire survey on training needs, and focus group interviews, a preliminary hierarchical training curriculum system for neonatal nurses was developed. Two rounds of Delphi correspondence were conducted with 19 domestic experts to finalize the system.Results:The effective questionnaire recovery rates of the two rounds of expert consultation were 95.00% and 100.00%, and the expert authority coefficient was 0.916, the Kendall harmony coefficient of the first round of expert opinions was 0.351 ( P<0.001), and the Kendall harmony coefficient of the second round of expert opinions was 0.463 ( P<0.001). The hierarchical training course structure and course training content are formed, including N0: 3 first-level items, 9 second-level items, 80 third-level items, N1: 3 first-level items, 9 second-level items, 91 third-level items, N2: 3 first-level items, 9 second-level items, 86 third-level items, N3: 3 first-level items, 10 second-level items, 81 third-level items, N4: 3 first-level items, 10 second-level items, 76 third-level items. Conclusions:The hierarchical training course system for neonatal nurses based on the core competence of nurses is scientific and practical, which can provide a reference for the hierarchical training of neonatal nurses.

Objective:To investigate the numbers of peripheral blood CD4 +T cell subpopulations in patients with elderly-onset rheumatoid arthritis (EORA) and its clinical significance. Methods:A total of 188 patients with newly diagnosed RA in the department of rheumatology and immunology of the Second Hospital of Shanxi Medical University from January 2020 to December 2023 were collected, including 48 cases of EORA (age of onset: ≥60 years old), 140 cases of young-onset rheumatoid arthritis (YORA) (18 years old ≤ age of onset < 60 years old). Meanwhile, 151 healthy controls (HC) were collected, of which 31 persons aged 60-85 years were included as HC group 1 (HC 1) and 120 individuals aged 18-59 years were included as HC group 2 (HC 2). Peripheral blood CD4 +T lymphocyte subsets of these participants were assessed by flow cytometry. Differences between groups were analyzed using independent-samples t test, Mann-Whitney U test or χ2 test. Results:Compared with healthy individuals, the absolute counts and percentages of peripheral blood Treg cells in patients with EORA were significantly decreased [absolute counts: 32.65 (23.04, 47.73) cells/μl vs. 23.03 (15.28, 32.12) cells/μl, Z=-3.35, P=0.001; percentages: 5.12%(4.13%, 6.16%) vs. 3.72% (2.79%, 4.82%), Z=-4.10, P<0.001], while the Th17/Treg cell ratio was increased [0.16 (0.12, 0.29) vs. 0.26 (0.18, 0.46), Z=-2.94, P=0.003], the differences are all significant. There was a tendency with higher absolute counts and percentages of Treg [absolute counts: 23.03 (15.28, 32.12) cells/μl vs. 20.97 (14.01, 30.64) cells/μl, Z=-0.58, P=0.561; percentages: 3.72%(2.79%, 4.82%) vs. 3.38% (2.39%, 4.71%), Z=-1.06, P=0.287] and lower Th17/Treg ratios [0.26 (0.18, 0.46) vs. 0.27 (0.19, 0.46), Z=-0.32, P=0.751] in EORA when compared to patients with YORA, but no significant differences were observed. Conclusion:Patients with EORA also have the reduced numbers of peripheral blood Treg cells and immune imbalance of Th17/Treg, suggesting that immune imbalance or dysfunction caused by defects in Treg cell counts and/or function contributes to the development of EORA, and that targeting Treg cells may be a promising therapeutic strategy for EORA.

Anxiety disorder is a common emotional disorder,with its pathological mechanisms involving the imbalance of multiple neurotransmitter systems,especially the dysfunction of the gamma-aminobutyric acid(GABA)system.This article explored the TCM understanding of anxiety disorder,and summarized and organized the research on the regulation of GABA system in the treatment of anxiety disorders through TCM monomers and formulas from the aspects of terpenes,flavonoids,phenolic acids,lignans,cinnamic acids,saponins,etc.At present,the research mainly involved chrysin,baicalin,ginsenoside,Baihe Dihuang Decoction,Bentun Decoction,Suanzaoren Decoction,Chaihu Jia Longgu Muli Decoction,etc.The mechanism involved regulating the inflammatory response of the body,alleviating oxidative stress damage,restoring the hyperactive HPA axis and other action pathways,which could provide a reference for the related research of TCM anti anxiety treatment and the development of new drugs.

ObjectiveTo investigate the mechanism of Baihuan Xiaoyao Decoction （Xiaoyaosan added with Lilii Bulbus and Albiziae Cortex） in alleviating depression-like behaviors of juvenile rats by regulating the polarization of microglia. MethodSixty juvenile SD rats were randomized into normal control， model， fluoxetine， and low-， medium-， and high-dose （5.36， 10.71， 21.42 g·kg^-1， respectively） Baihuan Xiaoyao decoction groups. The rat model of juvenile depression was established by chronic unpredictable mild stress （CUMS）. The sucrose preference test （SPT） was carried out to examine the sucrose preference of rats. Forced swimming test （FST） was carried out to measure the immobility time of rats. The open field test （OFT） was conducted to measure the total distance， the central distance， the number of horizontal crossings， and the frequency of rearing. Morris water maze （MWM） was used to measure the escape latency and the number of crossing the platform. The immunofluorescence assay was employed to detect the expression of inducible nitric oxide synthase （iNOS， the polarization marker of M1 microglia） and CD206 （the polarization marker of M2 microglia）. Real-time polymerase chain reaction was employed to determine the mRNA levels of iNOS， CD206， pro-inflammatory cytokines ［tumor necrosis factor （TNF）-α， interleukin （IL）-1β， and IL-6］ and anti-inflammatory cytokines （IL-4 and IL-10） in the hippocampus. Western blotting was employed to determine the protein levels of iNOS and CD206 in the hippocampus. The levels of IL-4 and IL-6 in the hippocampus were detected by enzyme-linked immunosorbent assay. ResultCompared with the normal control group， the model rats showed a reduction in sucrose preference （P<0.05）， an increase in immobility time （P<0.05）， decreased motor and exploratory behaviors （P<0.05）， and weakened learning and spatial memory （P<0.05）. In addition， the model rats showed up-regulated mRNA and protein levels of iNOS and mRNA levels of IL-1β， IL-6， and TNF-α （P<0.05）. Compared with the model group， Baihuan Xiaoyao decoction increased the sucrose preference value （P<0.05）， shortened the immobility time （P<0.01）， increased the motor and exploratory behaviors （P<0.05）， and improved the learning and spatial memory （P<0.01）. Furthermore， the decoction down-regulated the positive expression and protein level of iNOS， lowered the levels of TNF-α， IL-1β， and IL-6 （P<0.01）， promoted the positive expression of CD206， and elevated the levels of IL-4 and IL-10 （P<0.01） in the hippocampus of the high dose group. Moreover， the high-dose Baihuan Xiaoyao decoction group had higher sucrose preference value （P<0.01）， shorter immobility time （P<0.01）， longer central distance （P<0.01）， stronger learning and spatial memory （P<0.01）， higher positive expression and protein level of iNOS （P<0.01）， lower levels of TNF-α， IL-1β， and IL-6 （P<0.05， P<0.01）， lower positive expression and mRNA level of iNOS （P<0.05）， and higher levels of IL-4 and IL-10 （P<0.05， P<0.01） than the fluoxetine group. ConclusionBaihuan Xiaoyao decoction can improve the depression-like behavior of juvenile rats by inhibiting the M1 polarization and promoting the M2 polarization of microglia in the hippocampus.

AIM:To explore the mechanism by which Chaijin-Jieyu-Anshen tablet(CJJY)-medicated serum prevents synaptic injury in rat anterior cingulate cortex(ACC)neurons using an in vitro depression model.METHODS:Cells(astrocytes,microglia and neurons)were isolated from the ACC of SD rats.The isolated cells were characterized by immunofluorescence staining.An in vitro depression model was developed using 1 mg/L lipopolysaccharide(LPS)com-bined with 200 μmol/L corticosterone(CORT).These cells were divided into control group,model group(CORT+LPS),glucocorticoid receptor(GR)blocker(GR-)group(CORT+LPS+RU486),GR agonist(GR+)group(CORT+LPS+dexa-methasone),CX3C chemokine receptor 1(CX3CR1)blocker(CX3-)group(CORT+LPS+AZD8797),CX3CR1 agonist(CX3+)group(CORT+LPS+fractalkine),CJJY group(CORT+LPS+CJJY-medicated serum),CJJY/GR+group(CORT+LPS+CJJY-medicated serum+dexamethasone),and CJJY/CX3+group(CORT+LPS+CJJY-mediated serum+fractalkine).The morphological characteristics of all ACC cells were observed by high-content analysis.The levels of neuroendocrine-related factors,adrenocorticotropic hormone(ACTH),corticotropin-releasing hormone(CRH)and CORT,and neuroin-flammatory mediators,tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-6 and glutamate(Glu),in the cell supernatants were quantified by ELISA.Immunofluorescence staining was used to analyze the protein expression of GR and vesicular glutamate transporter 1(VGluT1)in astrocytes,as well as CX3CR1 and adenosine A2A receptor(A2AR)in microglia.Immunofluorescence staining with Nissl and β-tubulin was performed to evaluate synaptic damage in ACC neurons.RESULTS:In an in vitro model of depression,CJJY-medicated serum prevented morphological damage to ACC neurons,microglia and astrocytes.Moreover,CJJY-medicated serum reversed abnormal increases in the levels of ACTH,CRH,CORT,TNF-α,IL-1β,IL-6 and Glu in cell supernatants(P<0.05 or P<0.01).It was also found that CJJY-medi-cated serum reduced abnormal expression of GR,VGluT1,CX3CR1 and A2AR(P<0.05 or P<0.01),alleviating damage to the neuronal dendrites and dendritic spines of ACC neurons.CONCLUSION:The CJJY-medicated serum regulates GR/CX3CR1 double signaling in glia,and attenuates rat ACC neuronal synaptic damage in an in vitro depression model,indicating that CJJY-medicated serum controls depression by affecting the GR/CX3CR1 double signaling.