Aim To investigate the antidepressant mechanism of hyperforin via the utilization of single-cell sequencing technology.Methods C57BL/6 mice were randomly divided into the control group,depres-sion model group,and hyperforin intervention group.The chronic unpredictable mild stress(CUMS)model was induced and drug interventions were administered for 28 d.Behavioral experiments were conducted to as-sess depressive symptoms,and hippocampal tissue was collected for single-cell RNA sequencing.Key cell populations and differentially expressed genes across groups were identified,followed by PPI network,GO,and KEGG enrichment analysis.Results Behavioral experiments indicated that CUMS successfully induced depressive symptoms in mice,while hyperforin im-proved depressive behavior.In the depression model group,the proportion of brain perivascular macrophages(PVM)increased,and this proportion decreased after hyperforin intervention,approaching the level seen in the control group.The top 20 common differentially ex-pressed genes in the PVM subpopulation were Saa3,Hbb-bs and Ccl24.PPI network analysis identified core targets,including Ccl2,Dhx9,C3,Msr1,Cxcl2 and Cx3cr1.KEGG enrichment analysis revealed pathways related to chemokines,phagosome formation,and inosi-tol phosphate metabolism.Conclusion The antide-pressant mechanism of hyperforin may be related to the regulation of Ccl24 and its related chemokine signaling pathway by PVM.

Aim To investigate the antidepressant mechanism of hyperforin via the utilization of single-cell sequencing technology.Methods C57BL/6 mice were randomly divided into the control group,depres-sion model group,and hyperforin intervention group.The chronic unpredictable mild stress(CUMS)model was induced and drug interventions were administered for 28 d.Behavioral experiments were conducted to as-sess depressive symptoms,and hippocampal tissue was collected for single-cell RNA sequencing.Key cell populations and differentially expressed genes across groups were identified,followed by PPI network,GO,and KEGG enrichment analysis.Results Behavioral experiments indicated that CUMS successfully induced depressive symptoms in mice,while hyperforin im-proved depressive behavior.In the depression model group,the proportion of brain perivascular macrophages(PVM)increased,and this proportion decreased after hyperforin intervention,approaching the level seen in the control group.The top 20 common differentially ex-pressed genes in the PVM subpopulation were Saa3,Hbb-bs and Ccl24.PPI network analysis identified core targets,including Ccl2,Dhx9,C3,Msr1,Cxcl2 and Cx3cr1.KEGG enrichment analysis revealed pathways related to chemokines,phagosome formation,and inosi-tol phosphate metabolism.Conclusion The antide-pressant mechanism of hyperforin may be related to the regulation of Ccl24 and its related chemokine signaling pathway by PVM.

Aim To explore the mechanism of Con-grong Shujing granule(CSGs)in the treatment of Par-kinson's disease(PD)by network pharmacology,mo-lecular docking and parallel reaction monitoring(PRM)technology.Methods The active components of CSGs and the target genes of Parkinson's disease were obtained through the database.The intersection targets of drugs and diseases were selected to construct the"drug-active ingredient-target"and protein interac-tion network.The intersection target genes were impor-ted into David database for GO and KEGG enrichment analysis,and the main components were docked with key targets.27 SD rats were randomly divided into the normal group(n=9),model group(n=9)and treat-ment group(n=9).On day 1,7 and 14 of treatment,PRM analysis was used to detect the changes in the specific peptides of key target proteins in the substantia nigra of rats.Results The main components of CSGs wereTanshialdehyde,Baicalein,Quercetin and Kaempferol.The most important targets for the treat-ment of PD were TP53,AKT1,EGFR,HSP90 AA1 and STAT3.KEGG analysis mainly enriched MAPK,PI3K-Akt and neurotrophic factor signaling pathway.The molecular docking between core components and core targets showed that the binding of drugs and targets had good activity.PRM analysis of key proteins found that the target peptide expression levels of ASK1,JNK1 and JNK3 were different among groups(P＜0.05).Con-clusion CSGs can alleviate ERS,inhibit apoptosis and play a neural protective role through the ASK1-JNK pathway.

Objective To observe the clinical efficacy of multi-target repetitive transcranial magnetic stimulation on freezing of gait in Parkinson's disease. Methods A total of 45 Parkinson's disease patients with freezing of gait in our hospital from January 2023 to January 2024 were selected and randomly divided into primary motor cortex(M1) treatment group (M1 group),dorsolateral prefrontal cortex (DLPFC) group (D group),and M1+DLPFC group (MD group),with 15 cases in each group. The stimulation site of patients in M1 group was bilateral M1 region lower limb innervation region,the stimulation site of patients in D group was bilateral DLPFC,and the left M1 region and DLPFC region of patients in MD group were stimulated on alternate days. The efficacy of Parkinson's disease patients with freezing of gait and the changes of mood scores were compared among the groups. Results One patient in D group and one in MD group were lost to follow-up,and 43 patients were eventually included. The freezing of gait questionnaire (FOGQ) scores,timed up and go test (TUGT) total time,modified standing-start 180° turn test (SS-180) time,unified Parkinson's disease rating scale part Ⅲ(UPDRS Ⅲ) scores,Hamilton depression scale (HAMD) scores and Hamilton anxiety scale (HAMA) scores of patients after treatment were improved compared with those before treatment in all three groups (P<0.05),and the improvement effect of MD group was significantly better than those of M1 group and D group (P<0.05). Conclusion High-frequency repetitive transcranial magnetic stimulation targeting bilateral M1 region and DLPFC region can improve freezing of gait,walking ability,and motor function,and alleviate symptoms of depression and anxiety of Parkinson's disease patients.

Objective:To establish a stable rat model of sequelae of pelvic inflammatory disease(SPID)with clinical characteristics,and to provide a reliable experimental model for the study of the pharmcological effect and mechanism of SPID.Methods:Twenty-four 7-week-old SD rats were divided into sham operation group,model-A(108 cfu/mL mixed bacterial solution,0.2 mL),model-B(109 cfu/mL mixed bacterial solution 0.2 mL),and model-C(108 cfu/mL E.coli 0.2 mL).The weight of the rat's uterine was weighed and the uterine index was calculated.The automatic hematology analyzer was used to detect the blood routine;hematoxylin-eosin staining(HE)and masson staining were used to detect uterine pathlogical changes in rats.Enzyme-linked immunosorbent assay(ELISA)was used to detect interleukin-1β(IL-1β),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in rat uterine tissue homogenates.Western blot was used to detect the expression of proteins related to NF-κB signaling pathway.Results:Compared with the sham operation group,the uterine index of model-A,model-B,and model-C were significantly increased(P＜0.05,P＜0.01).The levels of WBC and NE in the model-A increased significantly(P＜0.01).The level of LY in model-B decreased significantly(P＜0.01).The levels of IL-1β,TNF-α in model-A,model-B,and model-C were significantly increased(P＜0.01).The levels of IL-6 in model-A and model-B were significantly increased(P＜0.05,P＜0.01).The collagen volume fraction of model-A and model-B were significantly increased(P＜0.01).Mechanism study indicates that the expression levels of p-IKKβ/IKKβ,p-IκBα/IκBα and p-p65/p65 in model-A were significantly increased(P＜0.01),and the expression levels of IκBα/β-actin were significantly decreased(P＜0.01).The expression level of p-IKKβ/IKKβ in model-B was significantly increased(P＜0.01).Conclusions:A stable rat model of SPID that conforms to clinical characteristics can be successfully constructed by combining 0.2 mL of mixed bacterial solution with a concentration of 108 cfu/mL and mechanical injury.This modeling method intervened in the expression of the NF-κB inflammatory signaling pathway.

The aim of this investigation was to determine the optimal storage medium for testicular hypothermic transportation and identify the ideal concentration for the application of the protective agent 5-aminolevulinic acid (5-ALA). Furthermore, this study aimed to explore the underlying mechanism of the protective effects of 5-ALA. First, we collected and stored mouse testicular fragments in different media, including Hank's balanced salt solution (HBSS; n = 5), Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F12; n = 5), and alpha-minimum essential medium (αMEM; n = 5). Storage of testicular tissue in HBSS preserved the integrity of testicular morphology better than that in the DMEM/F12 group ( P < 0.05) and the αMEM group ( P < 0.01). Testicular fragments were subsequently placed in HBSS with various concentrations of 5-ALA (0 [control], 1 mmol l -1 , 2 mmol l -1 , and 5 mmol l -1 ) to determine the most effective concentration of 5-ALA. The 2 mmol l -1 5-ALA group ( n = 3) presented the highest positive rate of spermatogonial stem cells compared with those in the control, 1 mmol l -1 , and 5 mmol l -1 5-ALA groups. Finally, the tissue fragments were preserved in HBSS with control ( n = 3) and 2 mmol l -1 5-ALA ( n = 3) under low-temperature conditions. A comparative analysis was performed against fresh testes ( n = 3) to elucidate the underlying mechanism of 5-ALA. Gene set enrichment analysis (GSEA) for WikiPathways revealed that the p38 mitogen-activated protein kinase (MAPK) signaling pathway was downregulated in the 2 mmol l -1 5-ALA group compared with that in the control group (normalized enrichment score [NES] = -1.57, false discovery rate [FDR] = 0.229, and P = 0.019). In conclusion, these data suggest that using 2 mmol l -1 5-ALA in HBSS effectively protected the viability of spermatogonial stem cells upon hypothermic transportation.
Male ; Animals ; Testis/cytology* ; Aminolevulinic Acid/pharmacology* ; Mice ; Organ Preservation/methods* ; Organ Preservation Solutions/pharmacology* ; Cryopreservation/methods*

Objective:To investigate the risk factors for postoperative coronal imbalance in patients with type A degenerative scoliosis and to refine the Qiu classification by reclassifying type A patients.Methods:A retrospective analysis was conducted on the clinical data of patients with type A degenerative scoliosis classified by the Qiu classification who underwent corrective surgery at our hospital from January 2017 to April 2022. A total of 69 patients were enrolled in the study, including 3 males and 66 females, with an age of 60.6±6.8 years (range: 47-71 years). Based on the preoperative CBD, type A patients were further divided into three subtypes: Aa with CBD≤1 cm, Ab with CBD>1 cm with C 7 plumb line deviation toward the concave side, and Ac with CBD>1 cm with C 7 plumb line deviation toward the convex side. The incidence of coronal imbalance was compared among subtypes. During follow-up, patients with CBD ≤3 cm were classified as balanced, and those with CBD>3 cm as imbalanced. Radiographic parameters including coronal Cobb's angle (CA), CBD, L 4 tilt angle, L 5 tilt angle, thoracic kyphosis (TK), lumbar lordosis (LL), pelvic incidence (PI), pelvic tilt (PT), sacral slope (SS), and sagittal vertical axis (SVA) were measured in the standing radiographs preoperatively, 2-weeks postoperatively and 2-years follow-up. Differences in radiographic parameters between balanced and imbalanced groups were compared, and binary logistic regression was used to identify risk factors for postoperative coronal imbalance. Results:In the imbalanced Group (22 patients), there were 6 patients of type Aa (27%), 5 patients of type Ab (23%), and 11 patients of type Ac (50%), and in the balanced Group (47 patients), there were 20 patients of type Aa (43%), 18 patients of type Ab (38%), and 9 patients of type Ae (19%). The difference in subtype distribution between the two groups was significant (χ 2=6.939, P=0.029). The CBD in the imbalanced group was significantly larger than in the balanced group at the 2-years follow-up (4.18±0.86 cm vs. 1.31±0.78 cm, t=-13.813, P<0.001). The preoperative L 4 tilt angle was significantly larger in the imbalanced group compared to the balanced group (23.59°±6.86° vs. 17.64°±8.34°, t=-2.914, P=0.005). And the postoperative and 2-years follow-up L 4 tilt angles were also significantly larger than the balanced group (18.6°±5.5° vs. 11.5°±5.7°, t=-4.904, P<0.001; 18.0°±5.6° vs. 11.1°±5.7°, t=-4.691, P<0.001). Regression analysis identified the Ac type [ OR=3.937, 95% CI(1.07, 14.55), P=0.040] and excessive postoperative L 4 tilt angle [ OR=1.288, 95% CI(1.09, 1.52), P=0.003] as risk factors for postoperative coronal imbalance in type A patients. Conclusions:In the A-type of Qiu classification, patients with preoperative CBD>1 cm and trunk tilting towards the convex side (Ac type), as well as those with a large L 4 tilt angle after surgery, are more likely to experience coronal plane imbalance after surgery.

Objective:To investigate the difference of Roussouly ideal classification in predicting postoperative proximal junctional kyphosis (PJK) between adult degenerative spinal deformity patients with and without pelvic fixation and the potential reasons.Methods:From January 2017 to January 2020, a total of 95 patients (4 males, 91 females; with an average age of 62.03±6.30 years) with degenerative spinal deformities were retrospectively analyzed. There were 35 patients in the non-pelvic group (1 male, 34 females) and 60 patients in the pelvic group (3 males, 57 females). The radiographic parameters included coronal Cobb's angle (CA), pelvic incidence (PI), pelvic tilt (PT), sacral slope (SS), thoracic kyphosis (TK), lumbar lordosis (LL), sagittal vertical axis (SVA), T1 pelvic angle (TPA), and proximal junctional angle (PJA) were measured in the standing radiographs preoperatively, postoperatively at 2 weeks, and 2-year follow-up. Changes in PT and SS were calculated for patients at 2 weeks and the 2-year follow-up. Based on the revised Roussouly classification, 95 patients were classified into different types preoperatively, postoperatively at 2 weeks, and during the 2-year follow-up. Changes in the classification of patients were documented postoperatively at 2 weeks. Roussouly types were determined using preoperative pelvic parameters, and a match was defined when the 2-week postoperative classification aligned with the ideal type. The occurrence of PJK and the relationship with classification matching were recorded in the group. Independent t-tests were used for intergroup comparisons of radiographic parameters, and chi-square tests were employed to assess classification changes and predictive accuracy of the Roussouly classification. Results:Preoperative PT, TPA and SVA in non-pelvic group were significantly smaller than those in pelvic group, and preoperative SS and LL larger than those in pelvic group ( P<0.05). The changes of PT and SS in non-pelvic group were significantly lower than those in pelvic group 2 weeks after surgery ( P<0.05). The proportion of classification changes in the pelvic group was significantly higher than that in the non-pelvic group (60% vs. 34%, χ 2=5.847, P=0.016). Among the 95 patients, a total of 29 experienced PJK during the follow-up, with 3 cases progressing to PJF. The incidence of PJK in mismatched patients was 37% with no significant difference compared with matched patients (19%) (χ 2=3.357, P=0.067). In the sacral spine group of 60 patients, 22 experienced PJK, with 3 cases progressing to PJF. Among them, 19 patients with PJK had a classification mismatch with the ideal classification at 2 weeks postoperatively. The PJK incidence was significantly higher in mismatched patients (45%) compared to matched patients (17%) (χ 2=4.429, P=0.035). In the non-pelvic group, 7 patients developed PJK, with 3 mismatched cases. The PJK incidence in mismatched vs. matched patients was 18% vs. 22%, showing no significant difference (χ 2=0.114, P=0.735). Conclusions:For the patients with degenerative spinal deformity, pelvic fixation leads to a more complete restoration of the ideal Roussouly classification. Restoration of the Roussouly type in patients with pelvic fixation is a reliable predictor of postoperative PJK. However, in patients without pelvic fixation, the alignment with the ideal Roussouly classification does not significantly correlate with PJK development.

Objective:To evaluate the clinical outcomes of corrective surgery in patients with different subtypes of "Ω"-type severe scoliosis.Methods:A retrospective analysis was conducted on 79 patients with "Ω"-type severe scoliosis treated at Nanjing Drum Tower Hospital from August 2010 to July 2020. The cohort included 37 males and 42 females, with a mean age of 21.4±7.4 years (range, 8-52 years). The mean duration of preoperative halo traction was 72.6±27.5 days (range, 14-150 days). Etiologies included congenital scoliosis (33 cases), idiopathic scoliosis (27 cases), Marfan syndrome (9 cases), neurofibromatosis (8 cases), and neuromuscular disorders (2 cases). Based on the classification by Karikari et al., 58 patients were classified as type 2P, 13 as type 2D, and 8 as type 2PD. Outcome measures included coronal and kyphotic Cobb angles, the Scoliosis Research Society-22 (SRS-22) questionnaire, Oswestry disability index (ODI), and visual analog scale (VAS).Results:All surgical procedures were successfully completed, and all patients were followed up for an average of 27.5±3.9 months (range, 24 to 40 months). The duration of Halo gravity traction was 72.6±27.5 days (range, 14-150 days). In the 2D group, the changes after traction were 26.0°±12.5° for the coronal Cobb angle and 10.1°±7.9° for the kyphotic Cobb angle. In the 2PD group, the traction effect was 13.4°±5.7° for the coronal and 8.3°±5.9° for the kyphotic Cobb angle. In the 2P group, the preoperative coronal Cobb angle was 128.9°±29.1°, postoperative was 84.5°±24.5°, and at the last follow-up was 87.7°±25.0°, yielding a correction rate of 34%±12%. The kyphotic Cobb angle in this group changed from 112.9°±27.1° preoperatively to 77.6°±22.9° postoperatively, and 80.2°±22.8° at the final follow-up, corresponding to a correction rate of 30%±16%. In the 2D group, the coronal Cobb angle was 113.1°±19.9° preoperatively, 71.2°±16.3° postoperatively, and 73.8°±16.3° at the final follow-up, with a correction rate of 37%±11%. The kyphotic Cobb angle in this group was 87.2°±14.0° preoperatively, 61.6°±18.5° postoperatively, and 65.1°±18.5° at the final follow-up, with a correction rate of 31%±22%. In the 2PD group, the coronal Cobb angle improved from 119.6°±29.0° preoperatively to 78.3°±20.8° postoperatively, and 87.0°±23.0° at the last follow-up, corresponding to a correction rate of 35%±8%. The kyphotic Cobb angle in this group was 124.6°±16.8° preoperatively, 82.1°±19.9° postoperatively, and 90.9°±16.9° at the final follow-up, with a correction rate of 33%±16%. At the last follow-up, SRS-22 scores across all four domains had improved in all three groups compared to preoperative values. In the 2PD group, however, the differences in the pain and self-image domains before and after surgery were not statistically significant ( P>0.05), while improvements in the other domains were significant ( P<0.05). No statistically significant differences were observed among the three groups either preoperatively or at the final follow-up ( P>0.05). Conclusion:Combined Halo-gravity traction and spinal corrective surgery are effective in improving both radiographic and functional outcomes in patients with "Ω"-type severe scoliosis. Nonetheless, patients in the 2PD subtype demonstrate reduced traction responsiveness and relatively limited postoperative recovery compared to the 2P and 2D subtypes.

Heart and spleen deficiency syndrome is the most common syndrome type in patients with insomnia. Based on the theory of disease syndrome-combined animal model， this paper used multiple databases to search for the keywords "heart and spleen deficiency"， "insomnia"， "sleepless"， "disease syndrome-combined animal model"， "model evaluation"， etc. It selected the literature related to the animal model of insomnia with heart and spleen deficiency in the past 20 years to evaluate from the aspects of model establishment， modeling factors， syndrome model， disease model， macro characterization & macro characterization evaluation scale， micro indicators， etc. It is found that the existing animal model of insomnia with heart and spleen deficiency is not completely constructed by the method of disease syndrome combination of disease modeling factors and syndrome modeling factors. In the model using this method， the single establishment factor of heart and spleen deficiency does not conform to the clinical reality of disease， and the selection of the factors for the insomnia model is not closely related to or even separated from the syndrome performance. There is a problem of insufficient quantification of macro representation when the macro representation of the model replaces the symptoms related to heart and spleen deficiency syndrome and insomnia in an equivalent manner for macro representation evaluation， which can be improved according to the quantitative ideas and examples of the existing macro representation and macro representation evaluation scale. There are few specific indicators of heart and spleen deficiency syndrome in micro indicators. The micro research of heart and spleen deficiency syndrome and the essence of other traditional Chinese medicine （TCM） syndromes can be carried out by metabonomics and other technologies combined with the theory of corresponding prescription and syndrome， along the specific related ideas of "prescription and syndrome， treatment principle and selection of prescription， treatment principle and selection of acupoints， as well as therapeutic mechanism and syndrome essence". The future users and researchers of animal models of insomnia with heart and spleen deficiency can get improved methods and ideas through the shortcomings of animal models of heart and spleen deficiency listed in this paper and construct animal models of insomnia with heart and spleen deficiency that are more suitable for clinical practice， so as to establish a more perfect modeling method and evaluation system of disease syndrome-combined animal model.