1.Mechanism of electroacupuncture on microglial and cortical neurons in mice with amyotrophic lateral sclerosis model
Weijia ZHAO ; Jiawei ZENG ; Junyang LIU
Chinese Journal of Rehabilitation Medicine 2025;40(2):171-179
Objective:To observe the effect of electroacupuncture(EA)on microglia and cortical excitability in hSOD1G93A mice and to explore the potential central mechanisms of EA intervention to improve motor func-tion in amyotrophic lateral sclerosis(ALS)mice.Method:The present study includes 2 parts.① Sixteen hSOD1G93A mice were randomly divided into model group and EA group(n=8 per group).Eight hSOD1G93A-negative mice were used as the blank group.The EA group was applied to the Baihui,bilateral Tianzhu and bilateral Tianshu acupoints(10 minutes/session,5 sessions/week,for 3 weeks).After 3 weeks of EA,hind limb functional neurologic score and open field test were used to evaluate the changes in motor function of mice.The concentrations of IL-1β and IL-6 were de-tected by ELISA.Immunofluorescence staining and flow cytometry were employed to observe the rate of Iba-1 positive cells and cell phenotypes in the cerebral cortex.②Twenty-one hSOD1G93A mice and seven wild-type mice were included.The Chemical genetic excitatory virus[rAAV-CaMKIIa-hM3D(Gq)-EGFP-WPRE-hGH poly A]was injected into the primary motor cortex(M1)area of the mice.After 21 days,four mice(3 SOD1G93A and 1 wild-type)were randomly selected for fluorescent microscopy to confirm viral expression.The remaining hSOD1G93A mice were randomly divided into 3 groups:model,EA group,and chemogenetic excitatory virus(intraperitoneal injection of clozapine-N-oxide[CNO])combined with EA group,with 6 mice per group.The rate of c-Fos positive cells in the cerebral cortex was observed by immunofluorescence.The open field experiment was used to evaluate the changes in motor function of mice.Result:Compared with the blank group,the model group showed significantly reduced total distance in the open-field test and increased neurologic scores(P<0.01),elevated serum IL-1β and IL-6 levels,increased corti-cal Iba-1 and c-Fos expression(P<0.01),and higher proportions of M1 and M2 microglia(P<0.01,P<0.05).Compared with the model group,the total distance of open field movement was prolonged and the neurologi-cal score decreased in the EA group(P<0.01,P<0.05).The contents of serum IL-1β and IL-6,the expression of Iba-1、c-Fos in cerebral cortex were decreased in the EA group(P<0.01,P<0.05),with an increased propor-tion of M2 microglia(P<0.05);the rate of c-Fos positive cells decreased in the virus+EA group,and the total distance of open field movement was prolonged.Conclusion:EA intervention can improve motor function in ALS mice.The underlying mechanism may be re-lated to suppress the activity of microglia,reduce the expression of inflammatory cytokines and further to de-crease the excitability of M1 cortical neurons.
2.A multi-center retrospective study on the effect of domestic RECO flow restoration device in acute ischemic stroke
Junyang YIN ; Qiyan ZONG ; Da WU ; Penghua LYU ; Yasuo DING ; Yan LIU ; Xiangzhong SHAO ; Guibing DING ; Yanbo CHENG ; Jie CAO ; Ya PENG
Chinese Journal of Cerebrovascular Diseases 2025;22(3):168-177
Objective To investigate the efficacy and safety of domestic RECO flow restoration device in endovascular treatment of acute ischemic stroke(AIS)patients in a real-world setting.Methods From January 2019 to June 2021,consecutive patients with acute intracranial vessel occlusion treated with RECO device from 8 stroke centers in Jiangsu Province were analyzed retrospectively.Baseline and clinical data were collected,including age,gender,baseline modified Rankin scale(mRS)scores,National Institutes of Health stroke scale(NIHSS)score on admission,trial of Org 10172 in acute stroke treatment(TOAST)classification,responsible location of occlusion,wake-up stroke,and medical history(hypertension,diabetes mellitus,hyperlipidemia,atrial fibrillation,coronary heart disease,peripheral arterial disease),smoking,and alcohol consumption.Furthermore,surgical parameters were collected,including time indicators(intervals between symptom onset,admission,puncture and recanalization),preoperative intravenous thrombolysis,general anesthesia,combination of aspiration,number of passes,first pass effect(modified thrombolysis in cerebral infarction[mTICI]grade 3 achieved with s single stent-based thrombectomy),rescue therapy(including thrombectomy with other stents,intra-arterial urokinase thrombolysis,intra-arterial application of tirofiban,emergency angioplasty[stent implantation,balloon dilatation,balloon dilatation combined with stent implantation]),balloon guided catheter and NIHSS score at discharge.The effectiveness indicators included immediate successful recanalization after the operation(mTICI grade ≥ 2b),complete recanalization(mTICI grade 3),and a good prognosis at 90 d after the operation(mRS score ≤2).The safety indicators included symptomatic intracranial hemorrhage(sICH)within 24 h after the operation,device-related intraoperative complications(such as thrombus escape,iatrogenic dissection,bleeding at the surgical site),and all-cause mortality within 90 days after the operation.All patients were divided into the anterior circulation occlusion group and the posterior circulation occlusion group according to the location of the responsible occluded vessel,and the effectiveness and safety indexes were analyzed.Results A total of 366 patients with AIS caused by intracranial vessel occlusion who underwent mechanical thrombectomy with the domestic RECO thrombectomy stent were included.The average age was(69±11)years,with 54 patients(14.8%)over 80 years old and 221 patients(60.4%)male.There were 296 patients in the anterior circulation occlusion group and 70 patients in the posterior circulation occlusion group.(1)Immediate successful recanalization(mTICI grade≥2b)was achieved in 337 patients(92.1%),and complete recanalization was achieved in 282 patients(77.0%)immediately after the operation.The average number of thrombectomies was(1.9±1.0)times,and 141 patients(38.5%)achieved recanalization on the first attempt.9 patients(2.5%)used other stents during the operation,and 65 patients(17.8%)underwent emergency angioplasty.The incidence of sICH within 24 h after the operation was 10.9%(40/366),the good prognosis rate at 90 d after the operation was 53.6%(196/366),and the all-cause mortality rate at 90 d after the operation was 23.0%(84/366).The incidence of device-related intraoperative complications was 3.6%(13/366).The median time from arterial puncture to recanalization was 75(52,110)min,and the median time from onset to recanalization was 370(280,488)min.(2)The good prognosis rates at 90 d after the operation in the anterior and posterior circulation occlusion groups were 55.1%(163/296)and 47.1%(33/70),respectively.The immediate successful recanalization rates were 93.2%(276/296)and 87.1%(61/70),respectively.The immediate complete recanalization rates were 79.1%(234/296)and 68.6%(48/70),respectively.The incidence of sICH within 24h after the operation was 12.2%(36/296)and 5.7%(4/70),respectively.The incidence of device-related intraoperative complications was 3.7%(11/296)and 2.9%(2/70),respectively.The all-cause mortality rate within 90 d after the operation was 21.6%(64/296)and 28.6%(20/70),respectively.There were no statistically significant differences between the two groups in the above aspects(all P>0.05).Conclusions The application of the domestic RECO thrombectomy stent in the treatment of AIS caused by intracranial vessel occlusion can effectively and safely achieve vascular recanalization.The results of this study still need to be further verified by prospective controlled studies.
3.Correlation of Serum sTREM-1 mRNA and HMGB1 mRNA Levels with Intestinal Flora and the Preterm Premature Rupture of Membranes in Gestational Diabe-tes Mellitus Patients
Junyang CAO ; Chunxing MA ; Zhina LIU
Journal of Practical Obstetrics and Gynecology 2025;41(6):501-507
Objective:To explore the relationship between serum soluble myeloid cell trigger receptor-1(sTREM-1)messenger RNA(mRNA),high mobility group protein B1(HMGB1)mRNA levels and intestinal flora and the occurrence of preterm premature rupture of membranes(PPROM)in gestational diabetes mellitus(GDM)patients.Methods:98 GDM pregnant women with GDM combined with PPROM who underwent regular prenatal check ups and delivered at the First Hospital Affiliated to Hebei North University from June 2022 to May 2024 were selected as the GDM combined with PPROM group,102 GDM pregnant women without PPROM were selected as the GDM without PPROM group,and 108 normal pregnant women were selected as the control group.Intestinal flora,serum sTREM-1 mRNA,HMGB1 mRNA levels of the three groups of pregnant women were collected and compared;the correlation between serum sTREM-1 mRNA and HMGB1 mRNA levels and intestinal flora in GDM pregnant women,the factors affecting the occurrence of PPROM in pregnant women with GDM,and the predictive value of serum sTREM-1 mRNA and HMGB1 mRNA in the occurrence of PPROM in GDM pregnant women were analyzed.Results:Compared with control group,colony number of Enterococcus,Enterobacter,Bacteroides and Fusobacterium were significantly increased in GDM without PPROM group and GDM combined with PPROM group(P<0.05),the colony number of Bifidobacterium and Lactobacillus was significantly de-creased(P<0.05).The gestational age at delivery of GDM combined with PPROM group was significantly lower than that of control group and GDM without PPROM group(P<0.05).Compared with GDM without PPROM group,the colony number of Bifidobacterium and Lactobacillus were significantly decreased(P<0.05),colony number of Enterococcus,Enterobacter,Bacteroides and Fusobacterium were significantly increased in GDM com-bined with PPROM group(P<0.05).The serum levels of sTREM-1 mRNA and HMGB1 mRNA in control group,GDM without PPROM group,GDM combined with PPROM group were increased sequentially(P<0.05).Serum sTREM-1 mRNA was positively correlated with HMGB1 mRNA in GDM pregnant women(r>0,P<0.05);serum sTREM-1 mRNA and HMGB1 mRNA were positively correlated with the colony numbers of Enterococcus,Enter-obacter,Bacteroides and Fusobacterium(r>0,P<0.05),and negatively correlated with the colony numbers of Bifidobacterium and Lactobacillus(r<0,P<0.05).Enterococcus,Enterobacter,Bacteroides,Fusobacterium,sTREM-1 mRNA and HMGB1 mRNA were independent risk factors for PPROM in GDM pregnant women(OR>1,P<0.05),while Bifidobacterium and Lactobacillus were independent protective factors for PPROM in GDM pregnant women(OR<1,P<0.05).The area under curve(AUC)of predicting PPROM in GDM pregnant women by combining serum sTREM-1 mRNA and HMGB1 mRNA(0.922)was higher than the AUC predicted by serum sTREM-1 mRNA and HMGB1 mRNA alone(0.829,0.841)(P<0.05).Conclusions:Serum sTREM-1 mRNA and HMGB1 mRNA were highly expressed in GDM pregnant women,and the increase trend of serum both was more obvious in GDM pregnant women with PPROM,both of which were related to intestinal flora imbalance and the occurrence of PPROM.The combination of sTREM-1 mRNA and HMGB1 mRNA has high efficacy in predicting the occurrence of PPROM.
4.Mechanism of electroacupuncture on microglial and cortical neurons in mice with amyotrophic lateral sclerosis model
Weijia ZHAO ; Jiawei ZENG ; Junyang LIU
Chinese Journal of Rehabilitation Medicine 2025;40(2):171-179
Objective:To observe the effect of electroacupuncture(EA)on microglia and cortical excitability in hSOD1G93A mice and to explore the potential central mechanisms of EA intervention to improve motor func-tion in amyotrophic lateral sclerosis(ALS)mice.Method:The present study includes 2 parts.① Sixteen hSOD1G93A mice were randomly divided into model group and EA group(n=8 per group).Eight hSOD1G93A-negative mice were used as the blank group.The EA group was applied to the Baihui,bilateral Tianzhu and bilateral Tianshu acupoints(10 minutes/session,5 sessions/week,for 3 weeks).After 3 weeks of EA,hind limb functional neurologic score and open field test were used to evaluate the changes in motor function of mice.The concentrations of IL-1β and IL-6 were de-tected by ELISA.Immunofluorescence staining and flow cytometry were employed to observe the rate of Iba-1 positive cells and cell phenotypes in the cerebral cortex.②Twenty-one hSOD1G93A mice and seven wild-type mice were included.The Chemical genetic excitatory virus[rAAV-CaMKIIa-hM3D(Gq)-EGFP-WPRE-hGH poly A]was injected into the primary motor cortex(M1)area of the mice.After 21 days,four mice(3 SOD1G93A and 1 wild-type)were randomly selected for fluorescent microscopy to confirm viral expression.The remaining hSOD1G93A mice were randomly divided into 3 groups:model,EA group,and chemogenetic excitatory virus(intraperitoneal injection of clozapine-N-oxide[CNO])combined with EA group,with 6 mice per group.The rate of c-Fos positive cells in the cerebral cortex was observed by immunofluorescence.The open field experiment was used to evaluate the changes in motor function of mice.Result:Compared with the blank group,the model group showed significantly reduced total distance in the open-field test and increased neurologic scores(P<0.01),elevated serum IL-1β and IL-6 levels,increased corti-cal Iba-1 and c-Fos expression(P<0.01),and higher proportions of M1 and M2 microglia(P<0.01,P<0.05).Compared with the model group,the total distance of open field movement was prolonged and the neurologi-cal score decreased in the EA group(P<0.01,P<0.05).The contents of serum IL-1β and IL-6,the expression of Iba-1、c-Fos in cerebral cortex were decreased in the EA group(P<0.01,P<0.05),with an increased propor-tion of M2 microglia(P<0.05);the rate of c-Fos positive cells decreased in the virus+EA group,and the total distance of open field movement was prolonged.Conclusion:EA intervention can improve motor function in ALS mice.The underlying mechanism may be re-lated to suppress the activity of microglia,reduce the expression of inflammatory cytokines and further to de-crease the excitability of M1 cortical neurons.
5.Value of blood lactic acid, procalcitonin, and total bilirubin in early diagnosis and prognosis evaluation of trauma complicated with sepsis
Jintao TANG ; Li HE ; Bangjia GAN ; Shijia CHAO ; Qinqin ZHANG ; Junyang MO ; Yujun LIU
Journal of Chinese Physician 2025;27(10):1478-1482
Objective:To explore the value of blood lactic acid (BLA), procalcitonin (PCT), and total bilirubin (TBil) in the diagnosis and prognosis evaluation of patients with trauma complicated with sepsis.Methods:The clinical data of 151 patients with severe trauma admitted to the Department of Emergency Medicine, Nanxishan Hospital of Guangxi Zhuang Autonomous Region from July 2019 to August 2023 were analyzed retrospectively. The patients were divided into the sepsis group (72 cases) and non-sepsis group (79 cases) according to the diagnosis. They were further divided into the death group (37 cases) and non-death group (114 cases) based on clinical outcomes. Clinical data were compared between groups. Receiver operating characteristic (ROC) curve was used to analyze the predictive efficacy of the above indicators, and Spearman correlation analysis was applied to evaluate the correlation between the indicators.Results:The levels of BLA, PCT, TBil, and Sequential Organ Failure Assessment (SOFA) score in the sepsis group were higher than those in the non-sepsis group (all P<0.05). The mortality rate of the sepsis group was significantly higher than that of the non-sepsis group, with a statistically significant difference [26/72(36.11%) vs 11/79(13.92%), χ 2=10.024, P=0.002]. The levels of BLA, PCT, TBil, and SOFA score in the death group were higher than those in the non-death group (all P<0.05). ROC curve analysis showed that the areas under the curve (AUC) of BLA, PCT, TBil, and their combination for diagnosing sepsis were 0.745, 0.826, 0.753, and 0.889 respectively; the sensitivity and specificity of the combined diagnosis of sepsis were 87.5% and 72.2%. The AUCs of BLA, PCT, TBil, and their combination for predicting the prognosis of sepsis were 0.644, 0.697, 0.614, and 0.713 respectively; the sensitivity and specificity of the combined prediction of sepsis prognosis were 64.9% and 71.1%. Among the 151 patients, the levels of BLA, PCT, TBil were positively correlated with SOFA score, with statistically significant differences ( r=0.3871, 0.4399, 0.4851, all P<0.001). Conclusions:BLA, PCT, and TBil levels have certain value in the early diagnosis and prognosis evaluation of patients with sepsis. The combined evaluation has the best efficacy and high guiding value in clinical practice.
6.A comparative study of the outcomes of primary reverse total shoulder arthroplasty and revision procedure after treatment failure of complex proximal humerus fractures in elderly patients
Bingshan YAN ; Ming XIANG ; Lei HAN ; Junyang LIU ; Bo ZHANG ; Xu TIAN ; Lintao LIU ; Jingming DONG
Chinese Journal of Orthopaedics 2025;45(15):984-992
Objective:To evaluate the clinical outcomesof primary reverse total shoulder arthroplasty (RTSA) and revision procedure with RTSA after treatment failure of complex proximal humeral fractures in the elderly.Methods:A retrospective analysis was conductedon 24 elderly patients with Neer three- or four-part proximal humeral fractures who underwent RTSA revision after treatment failure (RTSA revision group) from January 2017 to June 2022. There were 7 males and 17 females included, with a mean age of 78.23±5.78 years (range, 67-86 years). Forty-eight patients who underwent primary RTSA (primary RTSA group) during the same time period were selected by propensity score matchingin a 1∶2 ratio as controls, based on age, dominanthand, etiology, Neer typing, glenohumeral joint dislocation, rotator cuff integrity, and osteoporosis T-score. The primary RTSA group included 12 males and 36 females, with a mean age of 76.38±6.15 years (range, 65-87 years). Clinical indicators including demographic characteristics, healing rate of the greater tuberosity, visual analogue score (VAS), Constant-Murley score, American Shoulder and Elbow Surgeons (ASES), shoulder range of motion (ROM), patient satisfaction, and complication rate were collected and analyzed.Results:The mean follow-up duration was 40(32, 60) months (range, 25-72 months) in the primary RTSA group and 38(30, 61) months (range, 24-68 months) in RTSA revision group. There was no significant difference (χ 2=5.058, P=0.168) in the healing rate of the greater tuberosity between the primary RTSA group (41/48, 85.4%) and the RTSA revision group (15/24, 62.5%). Compared with preoperative status, the ROM of anterior elevation, abduction supination, external rotation, VAS score, Constant-Murley score, and ASES score were significantly improved at the last follow-up (all P<0.05) in the RTSA revision group. The anterior elevation (123.74°± 16.57°), abduction supination (113.73°±16.42°), and external rotation (36.45°±10.36°) in the primary RTSA group were superior to those in the RTSA revision group (109.43°±18.75°, 98.64°±15.47°, 30.47°±10.64°, respectively), the difference was statistically significant ( P<0.05). No statistical difference of ROM of internal rotation between the two groups was found (χ 2=4.034, P=0.133). At the last follow-up, the Constant-Murley scores (75.47±11.66) and ASES scores (73.58±15.72) of the primary RTSA group were higher than those in the RTSA revision group (60.43±10.24 and 63.28±18.77, respectively), and the differences were statistically significant ( P<0.05). In terms of VAS (1.66±0.93 vs. 2.02±1.15) and patient satisfaction [83%(40/48) vs. 88%(21/24)], no statistical difference was identified ( P>0.05). The complication rate were 10.4% (5/48) in the primary RTSA group and 20.8% (5/24) in the RTSA revision group (χ 2=1.452, P=0.285), with no serious complications requiring revision surgery in either group. Conclusions:For elderly patients with proximal humeral fractures after failed operation, RTSA revision might effectively improve the limb function and alleviatepain. However, compared with RTSA revision, primary RTSA demonstrated superiorearly clinical outcomes in shoulder ROM and functional recovery.
7.Correlation of Serum sTREM-1 mRNA and HMGB1 mRNA Levels with Intestinal Flora and the Preterm Premature Rupture of Membranes in Gestational Diabe-tes Mellitus Patients
Junyang CAO ; Chunxing MA ; Zhina LIU
Journal of Practical Obstetrics and Gynecology 2025;41(6):501-507
Objective:To explore the relationship between serum soluble myeloid cell trigger receptor-1(sTREM-1)messenger RNA(mRNA),high mobility group protein B1(HMGB1)mRNA levels and intestinal flora and the occurrence of preterm premature rupture of membranes(PPROM)in gestational diabetes mellitus(GDM)patients.Methods:98 GDM pregnant women with GDM combined with PPROM who underwent regular prenatal check ups and delivered at the First Hospital Affiliated to Hebei North University from June 2022 to May 2024 were selected as the GDM combined with PPROM group,102 GDM pregnant women without PPROM were selected as the GDM without PPROM group,and 108 normal pregnant women were selected as the control group.Intestinal flora,serum sTREM-1 mRNA,HMGB1 mRNA levels of the three groups of pregnant women were collected and compared;the correlation between serum sTREM-1 mRNA and HMGB1 mRNA levels and intestinal flora in GDM pregnant women,the factors affecting the occurrence of PPROM in pregnant women with GDM,and the predictive value of serum sTREM-1 mRNA and HMGB1 mRNA in the occurrence of PPROM in GDM pregnant women were analyzed.Results:Compared with control group,colony number of Enterococcus,Enterobacter,Bacteroides and Fusobacterium were significantly increased in GDM without PPROM group and GDM combined with PPROM group(P<0.05),the colony number of Bifidobacterium and Lactobacillus was significantly de-creased(P<0.05).The gestational age at delivery of GDM combined with PPROM group was significantly lower than that of control group and GDM without PPROM group(P<0.05).Compared with GDM without PPROM group,the colony number of Bifidobacterium and Lactobacillus were significantly decreased(P<0.05),colony number of Enterococcus,Enterobacter,Bacteroides and Fusobacterium were significantly increased in GDM com-bined with PPROM group(P<0.05).The serum levels of sTREM-1 mRNA and HMGB1 mRNA in control group,GDM without PPROM group,GDM combined with PPROM group were increased sequentially(P<0.05).Serum sTREM-1 mRNA was positively correlated with HMGB1 mRNA in GDM pregnant women(r>0,P<0.05);serum sTREM-1 mRNA and HMGB1 mRNA were positively correlated with the colony numbers of Enterococcus,Enter-obacter,Bacteroides and Fusobacterium(r>0,P<0.05),and negatively correlated with the colony numbers of Bifidobacterium and Lactobacillus(r<0,P<0.05).Enterococcus,Enterobacter,Bacteroides,Fusobacterium,sTREM-1 mRNA and HMGB1 mRNA were independent risk factors for PPROM in GDM pregnant women(OR>1,P<0.05),while Bifidobacterium and Lactobacillus were independent protective factors for PPROM in GDM pregnant women(OR<1,P<0.05).The area under curve(AUC)of predicting PPROM in GDM pregnant women by combining serum sTREM-1 mRNA and HMGB1 mRNA(0.922)was higher than the AUC predicted by serum sTREM-1 mRNA and HMGB1 mRNA alone(0.829,0.841)(P<0.05).Conclusions:Serum sTREM-1 mRNA and HMGB1 mRNA were highly expressed in GDM pregnant women,and the increase trend of serum both was more obvious in GDM pregnant women with PPROM,both of which were related to intestinal flora imbalance and the occurrence of PPROM.The combination of sTREM-1 mRNA and HMGB1 mRNA has high efficacy in predicting the occurrence of PPROM.
8.Artemin promotes proliferation and invasion of malignant peripheral nerve sheath tumor cells through the PI3K/Akt pathway
Hongliang ZHANG ; Haotian LIU ; Junyang LIU ; Chao ZHANG ; Ting LI ; Zhichao LIAO ; Yancheng LIU ; Jingyu ZHANG ; Kai ZHU ; Shuang LI ; Jinwei LIU ; Jilong YANG
Chinese Journal of Oncology 2025;47(2):149-159
Objective:To investigate the expression of Artemin (ARTN) in malignant peripheral nerve sheath tumor (MPNST), its effect on the malignant behavior of MPNST cells, and its signaling pathway.Methods:Fifty-one MPNST paraffin embedded tissues through surgical resection at Tianjin Medical University Cancer Hospital from January 1995 to November 2011 were collected, the expression of the ARTN protein was detected by immunohistochemistry, and the relationship between the ARTN protein expression and the clinical pathological characteristics and prognosis were analyzed. In human MPNST cell lines ST-8814 (NF-1) and STS26T(sporadic), ARTN overexpression and low expression cell lines were constructed by transfecting ARTN overexpression plasmids and ARTN small interfering RNA (siRNA), respectively. The expression of ARTN mRNA was detected by real time quantitative polymerase chain reaction (RT-qPCR), the expression of the ARTN protein and Phosphoinositide 3-kinase(PI3K)/Akt signaling pathway related proteins were detected by Western blot. CCK-8 assay was used to detect cell proliferation ability, and cell invasion assay was used to detect cell invasion ability. The pathway proteins that interacted with ARTN were searched in the STRING database, and the functional pathways were clarified by Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The PI3K/Akt pathway specific inhibitor LY294002 was used to block the PI3K/Akt pathway of ST-8814 and STS26T cells to observe the changes in cell proliferation and invasion.Results:Among the 51 MPNST tissue specimens, 22 cases showed a high expression of the ARTN protein and 29 cases showed a low expression of the protein. Higher expressions of the ARTN protein was associated with larger tumor diameters and disease progression (recurrence or metastasis) (both P<0.05). The median disease-free survival (DFS) of patients with a low expression of the ARTN protein was 26.2 months, and the median overall survival (OS) was 66.9 months. The median DFS and median OS of patients with a high expression of the ARTN protein were 10.7 months and 53.8 months, respectively. The log rank test results showed that the progression free survival rate of patients with a high expression of the ARTN protein was worse than that of patients with a low expression ( P=0.027), but the difference in overall survival rate between the two groups was not statistically significant ( P=0.790), which was also confirmed by Cox regression analysis. The CCK-8 assay results showed that after 48 hours of transfection, the absorbance ( A) values of ST-8814 and STS26T cells in the ARTN overexpression group were 1.35±0.01 and 1.10±0.02, respectively, which were higher than those in the empty plasmid control group (1.05±0.01 and 0.78±0.01, both P<0.01), while the A values of ST-8814 and STS26T cells in the ARTN siRNA group were 0.35±0.01 and 0.61±0.01, respectively, which were lower than those in the control siRNA group (0.74±0.01 and 1.10±0.04, both P<0.01). The results of cell invasion assay showed that the number of transmembrane cells in ST-8814 and STS26T cells overexpressing ARTN was (29.67±2.08) and (31.67±2.08), respectively, which were higher than those in the empty plasmid control group [(20.00±1.00) and (24.33±1.15), both P<0.01]. The number of transmembrane cells in ST-8814 and STS26T cells in the ARTN siRNA group were (14.00±2.00) and (19.33±1.53), respectively, which were lower than those in the control siRNA group [(19.33±2.52) and (23.33±0.58), both P<0.05].The KEGG results showed that ARTN is associated with multiple tumor signaling pathways, especially the PI3K/Akt signaling pathway. Western blot results showed that overexpression of ARTN upregulated the expression of p-PI3K and p-Akt proteins in ST-8814 and STS26T cells (both P<0.01).After knocking down ARTN expression, the expression of p-PI3K and p-Akt proteins was significantly down regulated (both P<0.01). LY294002 could significantly inhibit the effect of ARTN overexpression on ST-8814 and STS26T cells after blocking the PI3K/Akt pathway. The A values of ST-8814 and STS26T cells in the ARTN overexpression+LY294002 group were 1.09±0.06 and 0.82±0.01, respectively, which were lower than those in the ARTN overexpression group (1.50±0.01 and 1.29±0.01, respectively, both P<0.01). The numbers of transmembrane cells in the cell invasion assay were 16.67±3.21 and 19.67±2.31, respectively, which were also lower than those in the ARTN overexpression group (29.67±2.08 and 31.67±2.08, respectively, both P<0.01). Conclusions:In MPNST, a high expression of the ARTN protein was associated with larger tumor size, disease progression, and worse DFS. ARTN promotes the proliferation and invasion of MPNST cells through the PI3K/Akt signaling pathway.
9.Effects of electroacupuncture on the HMGB1/TLR4 pathway and oligodendrocytes in the cerebral cortex of mice with amyotrophic lateral sclerosis
Jiawei ZENG ; Weijia ZHAO ; Junyang LIU ; Shanshan LIU ; Le LI ; Weixing FENG ; Yingqian ZHAO ; Qiang WANG ; Chao JIANG
Journal of Acupuncture and Tuina Science 2025;23(5):385-393
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.
10.Pathological high-risk factors and prognostic analysis of intraocular stage advanced retinoblastoma following enucleation
Xueqing BAI ; Nan ZHANG ; Wen LIU ; Fei LENG ; Junyang ZHAO ; Li LI ; Chengyue ZHANG
Chinese Journal of Ocular Fundus Diseases 2025;41(7):507-513
Objective:To observe the high-risk histopathological feature (HRF) and their correlation with prognosis in children with intraocular retinoblastoma (RB) in the intraocular stage after failed eyepreserving treatment and enucleation surgery.Methods:A single-center retrospective case study. From August 2018 to January 2023, 64 children (64 eyes) with advanced intraocular RB who were diagnosed in Department of Ophthalmology of Beijing Children's Hospital and underwent enucleation surgery after failed eye-preserving treatment were included in the study. The median follow-up time was 51 months. The gender of the children patients, the age of visit and enucleation, International Intraocular Retinoblastoma Classification (IIRC), the initial chemotherapy regimen (hereinafter referred to as "chemotherapy"), the time of enucleation surgery, pathological results, post-enucleation treatment methods and prognosis were collected. The Mann-Whitney U test was used for comparison between groups. Survival analysis was performed using the Kaplan-Meier method, and the log-rank test was used for comparison between groups. Results:Among 64 cases and 64 eyes, 37 were male and 27 were female. The age of seeking medical treatment was 20 (11-31) months. The age at which the surgery was performed was 29 (16-40) months. The number of eyes in IIRC stage D and E was 16 and 48 respectively. The initial chemotherapy regimens simply applied (hereinafter referred to as "alone") intravenous systemic chemotherapy (IVC) and ophthalmic artery infusion chemotherapy (IAC) in 40 cases and 11 cases, 13 cases of IVC+IAC. All patients with positive HRF received systemic adjuvant chemotherapy after surgery. There were 37 eyes (57.8%, 37/64) positive for HRF. There was no statistically significant difference in the positive rate of HRF between children in IIRC stage D and stage E ( χ2=0.021, P=0.884). Among the 37 eyes with HRF, the numbers of eyes with extensive choroidal invasion, posterior lamina cribrosa optic nerve invasion, scleral invasion and optic nerve stump involvement were 17 (45.9%, 17/37), 16 (43.2%, 16/37), 3 (8.1%, 3/37) and 3 (8.1%, 3/37), respectively. During the follow-up period, there were 5 cases (7.8%, 5/64) of extraocular metastasis of the tumor and death, all of which were stage E and had HRF. Among them, the initial treatment plan was IAC for 4 cases, one case of IVC. The survival rates of children among the IVC, IAC or IVC+IAC regimens were 97.5% (39/40), 63.6% (7/11), and 100.0% (13/13), respectively. The comparison of survival rates among different chemotherapy regimens showed statistically significant differences ( χ2=14.233, P<0.001). The results of survival analysis showed that the cumulative survival rate of those with extensive choroidal invasion, posterior lamina cribrosa optic nerve infiltration, and those who received IAC was significantly lower than that of those without extensive choroidal invasion, posterior lamina cribrosa optic nerve infiltration, and those who received IVC+IAC and IVC ( P<0.05). Conclusions:Eye-preserving treatment for children with advanced intraocular RB may increase the positive rate of HRF and the risk of extraocular metastasis. The IVC+IAC eye-protecting treatment plan can improve the survival rate of children.

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