OBJECTIVE:Most of the existing studies are based on traditional Meta-analysis to study the efficacy of single stimulation protocols such as repetitive transcranial magnetic stimulation and transcranial direct current stimulation on lower limb motor dysfunction in stroke patients,and it is not possible to clarify which stimulation protocol is the optimal choice.This study used network meta-analysis to systematically evaluate the clinical efficacy of different regimens of non-invasive brain stimulation in the treatment of lower limb motor dysfunction after stroke.METHODS:CNKI,WanFang,VIP,CBM,PubMed,Medline and Web of Science databases were searched to collect randomized controlled trials on different regimens of non-invasive brain stimulation for lower limb motor dysfunction after stroke from inception to October 1,2024.Data extraction was performed on the included studies.RevMan 5.4.1 software was used for traditional meta-analysis and the quality of the included studies was evaluated.Stata 17.0 software was used for network meta-analysis.RESULTS:(1)A total of 39 studies involving 2 920 patients were included,involving 6 treatment methods:conventional rehabilitation training,high-frequency repetitive transcranial magnetic stimulation,low-frequency repetitive transcranial magnetic stimulation,intermittent theta burst stimulation,continuous theta burst stimulation,and transcranial direct current stimulation.(2)The results of network meta-analysis showed that high-frequency repetitive transcranial magnetic stimulation,low-frequency repetitive transcranial magnetic stimulation,intermittent theta burst stimulation,and transcranial direct current stimulation were superior to conventional rehabilitation training in the Fugl-Meyer assessment for lower extremity motor function.(3)In terms of improving Berg balance scale score,high-frequency repetitive transcranial magnetic stimulation,low-frequency repetitive transcranial magnetic stimulation,and intermittent theta burst stimulation were significantly different from conventional rehabilitation training(P＜0.05),and there was a significant difference between high-frequency repetitive transcranial magnetic stimulation and intermittent theta burst stimulation(P＜0.05).(4)In improving modified Barthel index and Barthel index,high-frequency repetitive transcranial magnetic stimulation,low-frequency repetitive transcranial magnetic stimulation,intermittent theta burst stimulation,and transcranial direct current stimulation were superior to conventional rehabilitation training.(5)Under the cumulative ranking chart,high-frequency repetitive transcranial magnetic stimulation showed the best efficacy in Fugl-Meyer assessment for lower extremity motor function,Berg balance scale score,modified Barthel index and Barthel index,followed by low-frequency repetitive transcranial magnetic stimulation.CONCLUSION:Both high-and low-frequency repetitive transcranial magnetic stimulation can improve the lower limb motor function and balance function of patients with stroke,and can improve the activities of daily living of patients to varying degrees.Moreover,the effect of high-frequency repetitive transcranial magnetic stimulation is better than that of low-frequency repetitive transcranial magnetic stimulation.

OBJECTIVE:Most of the existing studies are based on traditional Meta-analysis to study the efficacy of single stimulation protocols such as repetitive transcranial magnetic stimulation and transcranial direct current stimulation on lower limb motor dysfunction in stroke patients,and it is not possible to clarify which stimulation protocol is the optimal choice.This study used network meta-analysis to systematically evaluate the clinical efficacy of different regimens of non-invasive brain stimulation in the treatment of lower limb motor dysfunction after stroke.METHODS:CNKI,WanFang,VIP,CBM,PubMed,Medline and Web of Science databases were searched to collect randomized controlled trials on different regimens of non-invasive brain stimulation for lower limb motor dysfunction after stroke from inception to October 1,2024.Data extraction was performed on the included studies.RevMan 5.4.1 software was used for traditional meta-analysis and the quality of the included studies was evaluated.Stata 17.0 software was used for network meta-analysis.RESULTS:(1)A total of 39 studies involving 2 920 patients were included,involving 6 treatment methods:conventional rehabilitation training,high-frequency repetitive transcranial magnetic stimulation,low-frequency repetitive transcranial magnetic stimulation,intermittent theta burst stimulation,continuous theta burst stimulation,and transcranial direct current stimulation.(2)The results of network meta-analysis showed that high-frequency repetitive transcranial magnetic stimulation,low-frequency repetitive transcranial magnetic stimulation,intermittent theta burst stimulation,and transcranial direct current stimulation were superior to conventional rehabilitation training in the Fugl-Meyer assessment for lower extremity motor function.(3)In terms of improving Berg balance scale score,high-frequency repetitive transcranial magnetic stimulation,low-frequency repetitive transcranial magnetic stimulation,and intermittent theta burst stimulation were significantly different from conventional rehabilitation training(P＜0.05),and there was a significant difference between high-frequency repetitive transcranial magnetic stimulation and intermittent theta burst stimulation(P＜0.05).(4)In improving modified Barthel index and Barthel index,high-frequency repetitive transcranial magnetic stimulation,low-frequency repetitive transcranial magnetic stimulation,intermittent theta burst stimulation,and transcranial direct current stimulation were superior to conventional rehabilitation training.(5)Under the cumulative ranking chart,high-frequency repetitive transcranial magnetic stimulation showed the best efficacy in Fugl-Meyer assessment for lower extremity motor function,Berg balance scale score,modified Barthel index and Barthel index,followed by low-frequency repetitive transcranial magnetic stimulation.CONCLUSION:Both high-and low-frequency repetitive transcranial magnetic stimulation can improve the lower limb motor function and balance function of patients with stroke,and can improve the activities of daily living of patients to varying degrees.Moreover,the effect of high-frequency repetitive transcranial magnetic stimulation is better than that of low-frequency repetitive transcranial magnetic stimulation.

OBJECTIVE To investigate the correlation of serum P-Tau protein and β-amyloid protein expression levels with obstructive sleep apnea syndrome(OSAS)and mild cognitive impairment(MCI)patients,and their diagnostic value.METHODS From December 2020 to December 2023,120 patients with OSAS admitted to Third Hospital of Shijiazhuang were collected as the case group.According to the diagnostic criteria for MCI,patients were grouped into OSAS without MCI group(40 cases)and OSAS with MCI group(80 cases).ELISA method was applied to detect the levels of serum P-Tau protein and β-amyloid protein.Spearman method was applied to analyze the correlation between serum P-Tau protein,β-amyloid protein,and MCI.Multivariate logistic regression was applied to analyze the influencing factors of OSAS patients with MCI.ROC curve was applied to evaluate the diagnostic efficacy of serum P-Tau protein and β-amyloid protein in OSAS patients with MCI.RESULTS The Montreal Cognitive Assessment(MoCA)score in the OSAS with MCI group was obviously lower than that in the OSAS without MCI group(P<0.05).The expression levels of P-Tau protein and β-amyloid protein in the OSAS with MCI group were obviously higher than those in the OSAS without MCI group(P<0.05).The expression levels of serum P-Tau protein and β-amyloid protein in OSAS patients were negatively correlated with MoCA score(r=-0.346,-0.565,P<0.001).Serum P-Tau protein and β-amyloid protein were risk factors for OSAS with MCI(P<0.05).The AUC of the expression levels of serum P-Tau protein,β-amyloid protein,and their combination for OSAS with MCI was 0.751,0.848,and 0.928,respectively.The combined evaluation of the two showed better results(Zcombination-P-Tau protein=4.102,P<0.001;Zcombination-β amyloid protein=2.147,P=0.032).CONCLUSION The expression of serum P-Tau protein and β-amyloid protein is upregulated in OSAS patients with MCI,they are risk factors for the development of MCI in OSAS patients.The combined detection of the two has higher diagnostic efficacy.

OBJECTIVES: To investigate the inhibitory effect of multimerin-2 (MMRN2) overexpression on growth and metastasis of cutaneous melanoma cells. METHODS: Clinical data of patients with cutaneous melanoma were obtained from the GEO database to compare MMRN2 expressions between normal and tumor tissues. A protein-protein interaction network was constructed using the STRING database, and the intersecting genes from GEPIA2.0 were subjected to GO and KEGG enrichment analysis. The prognostic relevance of MMRN2 expression level was assessed using Cox regression and "timeROC". The correlations of MMRN2 expression level with immune infiltration and angiogenesis-related genes were analyzed using GSCA database and the ssGSEA algorithm. Colony-forming assay, Transwell assay, and wound healing assay were used to examine the changes in proliferation and migration of cultured cutaneous melanoma cells following MMRN2 knockdown. In a mouse model bearing cutaneous melanoma xenograft, the effect of MMRN2 knockdown on vital organ pathologies, survival of the mice and GM-CSF, CXCL9, and TGF‑β1 protein expressions were analyzed. RESULTS: MMRN2 was significantly upregulated in metastatic cutaneous melanoma (P<0.001). Protein interaction network analysis identified 15 intersecting genes, which were enriched in endothelium development and cell-cell junctions. In patients with cutaneous melanoma, a high MMRN2 expression was correlated with a poor prognosis, an advanced T stage, a greater Breslow depth, and ulceration (P<0.05). MMRN2 expression level was strongly correlated with 24 immune cell types (P<0.001), fibroblasts, endothelial cells, and expressions of the pro-angiogenic genes (KCNJ8, SLCO2A1, NRP1, and COL3A1; P<0.001). In cultured B16F10 cells, MMRN2 knockdown significantly suppressed cell proliferation, migration and invasion and caused remo-deling of the immunosuppressive microenvironment. CONCLUSIONS MMRN2 overexpression drives progression of cutaneous melanoma by enhancing tumor metastasis, angiogenesis and immune evasion, highlighting its potential as a therapeutic target for melanomas.
Humans ; Melanoma/metabolism* ; Animals ; Cell Movement ; Prognosis ; Skin Neoplasms/metabolism* ; Mice ; Cell Proliferation ; Neoplasm Invasiveness ; Cell Line, Tumor ; Protein Interaction Maps

Objective:To investigate effect of esketamine combined with ultrasound-guided stellate ganglion block(SGB)on stress response and postoperative immune function in elderly patients undergoing colorectal resection.Methods:A retrospective analy-sis was conducted in clinical data from 70 cases of elderly patients undergoing elective colorectal resection in Xuzhou Hospital of Tradi-tional Chinese Medicine,Nanjing University of Chinese Medicine from September 2020 to April 2022,who were randomly divided into observation group(n=35)and control group(n=35),control group was treated with ultrasound-guided SGB,observation group was treated with esketamine combined ultrasound-guided SGB.Heart rate(HR),mean arterial pressure(MAP),product of heart rate and systolic blood pressure(RPP)and stress response levels[cortisol(CORT),SOD,MDA,IL-6,TNF-α,IL-1β]were compared between two groups at 5 min before SGB(t0),immediately after tracheal intubation(t1),1 h after operation(t2)and end of operation(t3).CD3+T,CD4+T,CD8+T and CD4+T/CD8+T at t0,t3,24 h after operation(t4)and 72 h after operation(t5)were compared,respec-tively.Recovery time,first exhaust time,bowel sound recovery time and first oral feeding time and incidence of adverse reactions during anesthesia were compared between two groups.Results:HR,MAP,RPP,CORT,SOD,MDA,IL-6,TNF-α,IL-1β,CD3+T,CD4+T,CD8+T and CD4+T/CD8+T at different time points in two groups were compared by repeated measures analysis of variance,and there were significant differences between groups and at different time points(P<0.05).Compared with control group,observation group had shorter recovery time,first exhaust time,bowel sound recovery time,first oral feeding time,postoperative time out of bed(P<0.05),and less intraoperative pain medication consumption,incidence of adverse reactions during anesthesia was reduced[48.57%(17/35)vs 25.71%(9/35),P<0.05].Conclusion:Esketamine combined with ultrasound-guided SGB in elderly patients undergoing enterectomy can promote recovery of postoperative gastrointestinal function,reduce perioperative stress response,has little impact on immune function of patients,reduce perioperative adverse reactions.

Objective:To compare the efficacy of Wnt3a factor and small molecule compound CHIR99021 in culturing colorectal cancer organoid, and to explore the feasibility of replacing Wnt3a with CHIR99021.Methods:The organoids were cultured using 2 culture systems containing Wnt3a or CHIR99021, based on the colorectal cancer cell line HCT116 and rectal cancer tissue from one patient (surgical specimen from the Department of Gastrointestinal Surgery, Zhongshan Hospital Affiliated to Dalian University), including Wnt3a cell organoid, CHIR99021 cell organoid, Wnt3a tissue organoid, and CHIR99021 tissue organoid. The growth of organoids was observed under the optical microscope. The pathological characteristics of organoids and the rectal cancer tissue were analyzed by hematoxylin-eosin (HE) staining and immunohistochemical (IHC) staining, which included cytokeratin(CK) 7, CK20, Ki-67, and stemness marker CD133. The expression of β-catenin (a key Wnt pathway protein) was analyzed by Western blotting (WB) method. The half maximal inhibitory concentration (IC50) values of Wnt3a and CHIR99021 cell organoids were analyzed by drug susceptible test and GraphPad Prism 9.0 software. Independent sample- t test was used for statistical analysis. Results:Under the optical microscope, the size of CHIR99021 cell organoid was relatively uniform, while the size of the Wnt3a cell organoid was uneven, compact and dense spherical structure was formed in both organoids. HE staining showed tumor features including increased nuclear-cytoplasmic ratio and obvious nuclear atypia in the Wnt3a and CHIR99021 cell organoids. The results of IHC staining showed that CK7 was negative, and CK20 and Ki-67 were positive in the Wnt3a and CHIR99021 cell organoids. The results of WB method showed that the relative expression level of β-catenin of the CHIR99021 cell organoid was higher than that of the Wnt3a cell organoid (0.89±0.09 vs. 0.26±0.04), and the difference was statistically significant ( t=13.80, P<0.001). The results of drug susceptible test demonstrated that the IC50 value of the Wnt3a and CHIR99021 cell organoid was 10.91 and 14.55 μmol/L, respectively. Further IHC staining showed that CD133 was positive in the Wnt3a and CHIR99021 cell organoids, with stronger intensity in the CHIR99021 cell organoid. The pathological characteristics of Wnt3a and CHIR99021 tissue organoid were consistent with those of the rectal cancer tissue of the patient, with all CK7 being negative and CK20 and Ki-67 being positive. Conclusions:Both Wnt3a and CHIR99021 can successfully establish colorectal cancer organoids with consistent pathological characteristics. The IC50 value of the CHIR99021 cell organoid is high, which is related to the increased stemness of organoids. The pathological characteristics of Wnt3a and CHIR99021 tissue organoid are consistent with those of the rectal cancer tissue from the patient.

Objective:To observe any therapeutic effect of repeated high-frequency transcranial magnetic stimulation (rTMS) on patients with medication overuse headache (MOH).Methods:Fifty-seven persons with MOH were divided at random into a control group of 29 cases and an rTMS group of 28 cases. Both groups of patients stopped taking analgesics and were treated with topiramate and trazodone. The rTMS group received an additional 20 minutes of rTMS treatment daily, 5 days a week for 8 consecutive weeks. The control group received sham stimulation. Before and after the treatment, both groups′ headache symptoms, disability, depression, anxiety, cognition and sleep quality were evaluated using a visual analog scale (VAS) for pain, a migraine disability assessment scale (MIDAS), the Montreal cognitive assessment (MoCA), the Hamilton depression scale (HAMD), the Hamilton anxiety scale (HAMA) and the Pittsburgh sleep quality index (PSQI). Auditory event-related potential P300 was also measured for both groups.Results:After the treatment, the average VAS, MIDAS, HAMD, HAMA and PSQI scores of both groups had decreased significantly. The HAMD, HAMA and PSQI average scores of the rTMS group were then significantly better than the control group′s averages. The average MoCA score of the rTMS group had also improved significantly. The average latency of the auditory event-related potential P300 in the rTMS group was then (368.25±9.25)ms and its amplitude was (6.62±1.40)μV, showing significant improvement compared to before the treatment and compared with the control group.Conclusions:On the basis of oral administration of topiramate and trazodone, additional high-frequency rTMS treatment can significantly alleviate anxiety and depression in MOH patients, and improve their sleep quality and the latency and amplitude of event-related potential P300.

Chronic obstructive pulmonary disease (COPD) is the fourth leading cause of death worldwide, causing a significant socioeconomic burden. This article reviews the effects of influenza vaccination on COPD and finds that influenza vaccine can significantly reduce the risk of influenza infection, reduce the number of acute exacerbations, and reduce the hospitalization rate in patients with COPD. The vaccine has a favorable safety profile and significant economic benefits, which can reduce medical costs. Currently, influenza vaccination mainly faces challenges such as insufficient patient awareness, insufficient support from the medical system, and socio-cultural and economic factors. Efforts should be focused on reducing the acute exacerbation of COPD patients and providing a scientific basis for the prevention and management of COPD patients.

Objective:To explore the effects of early electroacupuncture(EA)intervention on the high mobility group box 1(HMGB1)/Toll-like receptor 4(TLR4)signaling pathway-related protein expression and oligodendrocytes in mice with amyotrophic lateral sclerosis(ALS),and uncover the potential molecular mechanisms underlying the improvement of motor function in ALS mice by early EA intervention.Methods:ALS mice carrying the SOD1G93A gene were randomly divided into a model group and an EA group,with 10 mice in each group;10 littermate mice with a negative SOD1G93A genotype served as the control group.In the EA group,Baihui(GV20),Tianzhu(BL10),and Tianshu(ST25)were selected with needles retained for 10 min,5 consecutive days per week,with 2 days of rest.One week constituted a course of treatment,and a total of 3 consecutive courses were performed.The other groups were grasped and fixed similarly,but without intervention.Motor function was assessed using the open field test(OFT)and Morris water maze(MWM).Subsequently,hematoxylin-eosin staining was used to observe neuron morphology in the M1 region of the cerebral cortex.Immunofluorescence was performed to detect the positive cell rate of TAR DNA-binding protein 43(TDP-43),and double immunofluorescence staining was used to observe the positive cell rate and cell states of ionized calcium-binding adaptor molecule 1(Iba-1)and myelin basic protein(MBP)in the M1 region of the cerebral cortex.Western blotting was used to detect the relative expression levels of TDP-43,tumor necrosis factor(TNF)-α,HMGB1,and TLR4 proteins.Results:Compared to the control group,the model group exhibited a reduced total movement distance in the OFT,and an increased escape latency,as well as fewer platform crossings in the MWM,with statistically significant differences(P<0.01).In the model group,the number of degenerated and necrotic neurons in the M1 region of the ALS mouse cerebral cortex increased,with significant nuclear shrinkage and cytoplasmic vacuolization;the percentage of TDP-43 immunofluorescence positive cells in the M1 region of the cerebral cortex increased(P<0.01),and the relative expression level of TDP-43 protein in the cerebral cortex showed a significant increase(P<0.01);the Iba-1 positive cell percentage increased,while the MBP positive cell percentage decreased(P<0.01);the relative expression levels of TNF-α,HMGB1,and TLR4 proteins increased(P<0.05).Compared to the model group,the EA group showed an increased total movement distance(P<0.01),and a reduced escape latency,and more platform crossings in the MWM,with statistically significant differences(P<0.05).In the EA group,neurons showed improvement,with reduced degeneration and necrosis,and larger,clearer nuclei;the percentage of TDP-43 immunofluorescence positive cells in the M1 region of the cerebral cortex decreased(P<0.05),and the relative expression level of TDP-43 protein also decreased(P<0.05);the percentage of Iba-1 positive cells in the M1 region of the cerebral cortex decreased,while the percentage of MBP positive cells increased(P<0.01);the relative expression levels of TNF-α,HMGB1,and TLR4 proteins decreased(P<0.05).Conclusion:EA intervention can suppress microglial activation,improve the state of oligodendrocytes,and reduce abnormal TDP-43 aggregation in the M1 region of the cerebral cortex in ALS model mice;its mechanism of action may be related to the HMGB1/TLR4 signaling pathway.