Objective:To identify factors influencing treatment-free remission (TFR) outcomes in children and adolescent patients with chronic myeloid leukemia (CML) after imatinib (IM) discontinuation.Methods:This multicenter retrospective study analyzed 36 children and adolescent patients with CML from eight hematology centers in China (December 1, 2016, to September 27, 2024) who discontinued IM therapy with documented post-cessation outcomes. Clinical characteristics and molecular response dynamics were assessed. Univariate analysis and multivariate Cox proportional hazards regression models were employed to assess factors associated with TFR outcomes.Results:A total of 36 patients were documented, comprising 17 males and 19 females. The median ages at CML diagnosis and IM discontinuation were 11 years ( IQR: 5,16) and 20 years ( IQR: 14,25), respectively. The median time from IM initiation to first deep molecular response (DMR) was 21 months ( IQR: 13, 38). Pre-discontinuation, patients received IM for a median duration of 96 months ( IQR: 84, 121) and maintained DMR for 74 months ( IQR: 63, 89). With a median post-discontinuation follow-up of 38 months ( IQR: 15, 68), cumulative TFR rates at 6, 12, 24, and 36 months were 74.1%, 60.7%, 60.7%, and 56.0%, respectively, generating an overall TFR rate of 58.3%. Fifteen patients lost major molecular response at a median of 5 months post-discontinuation ( IQR: 3, 11). All 15 patients resumed tyrosine kinase inhibitor therapy, comprising 13 who restarted IM and 2 who switched to dasatinib. By the last follow-up, 13 (86.7% ) patients regained DMR after a median treatment duration of 5 months ( IQR: 3, 17), and no disease progression occurred in any patient. Withdrawal syndrome occurred in 2 (5.6% ) patients. Univariate analysis revealed significantly higher TFR rates in patients with pre-discontinuation IM duration of ≥100 months vs <100 months (82.4% vs 36.8%, P=0.017) and pre-discontinuation DMR duration of ≥72 months vs <72 months (84.2% vs 29.4%, P=0.003). Multivariate Cox analysis identified pre-discontinuation DMR duration as an independent protective factor for TFR ( HR=5.419, 95% CI: 1.524–19.272, P=0.009) . Conclusion:DMR duration was identified as an independent protective factor influencing TFR outcomes in children and adolescent patients with CML after IM discontinuation. Patients who maintained DMR for ≥72 months before IM discontinuation demonstrated a significantly higher TFR rate.

Multiple system atrophy （MSA） is a rare degenerative disease of the nervous system and has diverse and atypical clinical manifestations， and it overlaps with other diseases in α-synuclein spectrum disease. There are great challenges in the diagnosis and early differential diagnosis of the disease， and missed diagnosis and misdiagnosis occur from time to time， thereby delaying the treatment of the disease.Videonystagmography （VNG） is currently the main noninvasive test used to assess vestibular function and can provide different eye movement parameters. Studies have shown the presence of abnormal eye movements in patients with MSA. From the perspective of vision-eye movement， this article reviews the current status of research on eye movements in patients with MSA and reveals the internal connection between them， in order to provide an important reference for the early diagnosis of MSA.
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Objective:To identify factors influencing treatment-free remission (TFR) outcomes in children and adolescent patients with chronic myeloid leukemia (CML) after imatinib (IM) discontinuation.Methods:This multicenter retrospective study analyzed 36 children and adolescent patients with CML from eight hematology centers in China (December 1, 2016, to September 27, 2024) who discontinued IM therapy with documented post-cessation outcomes. Clinical characteristics and molecular response dynamics were assessed. Univariate analysis and multivariate Cox proportional hazards regression models were employed to assess factors associated with TFR outcomes.Results:A total of 36 patients were documented, comprising 17 males and 19 females. The median ages at CML diagnosis and IM discontinuation were 11 years ( IQR: 5,16) and 20 years ( IQR: 14,25), respectively. The median time from IM initiation to first deep molecular response (DMR) was 21 months ( IQR: 13, 38). Pre-discontinuation, patients received IM for a median duration of 96 months ( IQR: 84, 121) and maintained DMR for 74 months ( IQR: 63, 89). With a median post-discontinuation follow-up of 38 months ( IQR: 15, 68), cumulative TFR rates at 6, 12, 24, and 36 months were 74.1%, 60.7%, 60.7%, and 56.0%, respectively, generating an overall TFR rate of 58.3%. Fifteen patients lost major molecular response at a median of 5 months post-discontinuation ( IQR: 3, 11). All 15 patients resumed tyrosine kinase inhibitor therapy, comprising 13 who restarted IM and 2 who switched to dasatinib. By the last follow-up, 13 (86.7% ) patients regained DMR after a median treatment duration of 5 months ( IQR: 3, 17), and no disease progression occurred in any patient. Withdrawal syndrome occurred in 2 (5.6% ) patients. Univariate analysis revealed significantly higher TFR rates in patients with pre-discontinuation IM duration of ≥100 months vs <100 months (82.4% vs 36.8%, P=0.017) and pre-discontinuation DMR duration of ≥72 months vs <72 months (84.2% vs 29.4%, P=0.003). Multivariate Cox analysis identified pre-discontinuation DMR duration as an independent protective factor for TFR ( HR=5.419, 95% CI: 1.524–19.272, P=0.009) . Conclusion:DMR duration was identified as an independent protective factor influencing TFR outcomes in children and adolescent patients with CML after IM discontinuation. Patients who maintained DMR for ≥72 months before IM discontinuation demonstrated a significantly higher TFR rate.

Objective:To investigate the effects of insulin-like growth factor-binding protein 3(IGFBP3),which is carried in exosomes released by cisplatin(DDP)-tolerant human lung adenocarcinoma(LUAD)A549/DDP cells,on differentiation of macrophages and its effect on DDP resistance of A549 cells.Methods:The parental A549 and A549/DDP cells were cultured in vitro,and the IC50 values were calculated after treatment with different concentrations of DDP for 48 h.The supernatants of A549 or A549/DDP cells culture were collected,and the exosomes were isolated using ultracentrifugation and named A-exo or A/D-exo,respectively.THP-1 cells were induced to differentiate into M0-type macrophages with PMA(15 μg/mL),mixed with A549 cells at a ratio of 1∶1,and then inoculated in the axillae of nude mice;on the day of tumor cell inoculation,the tumor cells were injected with PBS,A-exo,and A/D-exo at the inoculation site of the tumor cells,respectively,and at the same time,the treatment was carried out by intraperitoneal injection of DDP 1 time every 4 d.On the 35th day of the tumor loading in mice,the recruitment of human CD11b+CD206+or CD11b+CD86+macrophages in transplanted tumor tissues was detected by flow cytometry(FCM).Antibody microarrays were used to screen for proteins carried by A-exo or A/D-exo and validated by detecting the amount of IGFBP3 protein in A-exo and A/D-exo by ELISA method.A549 or A549/DDP cells were treated with different concentrations of rhIGFBP3,and the effects of rhIGFBP3 on the proliferation or migration ability of the cells were detected by MTS assay and Transwell assay,respectively.M0-type macrophages were treated with rhIGFBP3 for 4 d,and the culture supernatant was collected;the effects of different concentrations of rhIGFBP3 on the production of TGF-β and TNF-α content by M0-type macrophages were detected by ELISA;in addition,A549 cells were treated with rhIGFBP3 or culture supernatant of M0-type macrophages pretreated with rhIGFBP3,and again detected the IC50 value of DDP on A549 cells.Results:The IC50 value of DDP on A549/DDP cells was significantly higher than that of A549 cells(P<0.01);A/D-exo significantly promoted the growth of A549 cells xenograft tumors(P<0.05)and facilitated the recruitment of CD11b+CD206+macrophages into tumor tissues(P<0.05),compared with PBS and A-exo groups.Exosomes A-exo and A/D-exo were successfully obtained;high levels of IGFBP3 were carried in A/D-exo compared with A-exo.The analysis showed that the expression level of IGFBP3 was significantly up-regulated in patients with LUAD,and the overall survival rate of patients with high expression of IGFBP3 was reduced compared with those with low expression of IGFBP3.High concentration of rhIGFBP3(100 ng/mL)had a significant pro-proliferative effect on either A549 or A549/DDP cells(both P<0.05),but there was no statistically significant effect on the migratory ability of A549 or A549/DDP cells.High concentrations of rhIGFBP3(100 ng/mL)induced TGF-β1 production by M0-type macrophages(P<0.05),but not TNF-α production.The IC50 value of DDP on A549 cells was significantly increased(P<0.05)after treatment with culture supernatant of M0-type macrophages pretreated with IGFBP3(but not rhIGFBP3).Conclusion:A549/DDP cells mediate M2-type macrophage differentiation and promote secondary drug resistance in A549 cells by secreting IGFBP3-rich exosomes.

Objective:To investigate the effects of insulin-like growth factor-binding protein 3(IGFBP3),which is carried in exosomes released by cisplatin(DDP)-tolerant human lung adenocarcinoma(LUAD)A549/DDP cells,on differentiation of macrophages and its effect on DDP resistance of A549 cells.Methods:The parental A549 and A549/DDP cells were cultured in vitro,and the IC50 values were calculated after treatment with different concentrations of DDP for 48 h.The supernatants of A549 or A549/DDP cells culture were collected,and the exosomes were isolated using ultracentrifugation and named A-exo or A/D-exo,respectively.THP-1 cells were induced to differentiate into M0-type macrophages with PMA(15 μg/mL),mixed with A549 cells at a ratio of 1∶1,and then inoculated in the axillae of nude mice;on the day of tumor cell inoculation,the tumor cells were injected with PBS,A-exo,and A/D-exo at the inoculation site of the tumor cells,respectively,and at the same time,the treatment was carried out by intraperitoneal injection of DDP 1 time every 4 d.On the 35th day of the tumor loading in mice,the recruitment of human CD11b+CD206+or CD11b+CD86+macrophages in transplanted tumor tissues was detected by flow cytometry(FCM).Antibody microarrays were used to screen for proteins carried by A-exo or A/D-exo and validated by detecting the amount of IGFBP3 protein in A-exo and A/D-exo by ELISA method.A549 or A549/DDP cells were treated with different concentrations of rhIGFBP3,and the effects of rhIGFBP3 on the proliferation or migration ability of the cells were detected by MTS assay and Transwell assay,respectively.M0-type macrophages were treated with rhIGFBP3 for 4 d,and the culture supernatant was collected;the effects of different concentrations of rhIGFBP3 on the production of TGF-β and TNF-α content by M0-type macrophages were detected by ELISA;in addition,A549 cells were treated with rhIGFBP3 or culture supernatant of M0-type macrophages pretreated with rhIGFBP3,and again detected the IC50 value of DDP on A549 cells.Results:The IC50 value of DDP on A549/DDP cells was significantly higher than that of A549 cells(P<0.01);A/D-exo significantly promoted the growth of A549 cells xenograft tumors(P<0.05)and facilitated the recruitment of CD11b+CD206+macrophages into tumor tissues(P<0.05),compared with PBS and A-exo groups.Exosomes A-exo and A/D-exo were successfully obtained;high levels of IGFBP3 were carried in A/D-exo compared with A-exo.The analysis showed that the expression level of IGFBP3 was significantly up-regulated in patients with LUAD,and the overall survival rate of patients with high expression of IGFBP3 was reduced compared with those with low expression of IGFBP3.High concentration of rhIGFBP3(100 ng/mL)had a significant pro-proliferative effect on either A549 or A549/DDP cells(both P<0.05),but there was no statistically significant effect on the migratory ability of A549 or A549/DDP cells.High concentrations of rhIGFBP3(100 ng/mL)induced TGF-β1 production by M0-type macrophages(P<0.05),but not TNF-α production.The IC50 value of DDP on A549 cells was significantly increased(P<0.05)after treatment with culture supernatant of M0-type macrophages pretreated with IGFBP3(but not rhIGFBP3).Conclusion:A549/DDP cells mediate M2-type macrophage differentiation and promote secondary drug resistance in A549 cells by secreting IGFBP3-rich exosomes.

Abstract OBJECTIVE To develop an ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for determination of hynchophylline, isorhynchophylline, isocorynoxeine, corynoxeine, hirsutine and hirsuteine in Uncaria decoction, and investigate the effect of different storage conditions on the stability of active ingredients. METHODS An UHPLC-MS/MS method was established to investigate the effects of different storage temperature and time on the stability of six active components in Uncaria decoction. Separation was performed with a gradient mobile phase system of acetonitrile-0.1% formic water solution, the flow rate was 0.4 mL·min-1, the temperature of column was 30℃, the injection volume was 2 μL, the MS detection was dynamic multiple reaction monitoring mode. The effects of different storage temperature and time on the stability of six active components in Uncaria decoction was investigated. RESULTS Rhynchophylline, isorhynchophylline, corynoxeine, isocorynoxeine, hirsutine and hirsuteine were successfully separated using this method, with good linear relationship in the corresponding concentration range. The precision, repeatability, stability and recovery rate were good. The six alkaloids were basically stable within 14 d under low temperature storage, basically stable within 3 d under normal temperature storage, and not stable under high temperature storage. CONCLUSION The UHPLC-MS/MS method is stable, rapid and reproducible. It can be used to determine the contents of six effective components in Uncaria decoction. Cold storage is beneficial to the stability of Uncaria decoction.

Objective To investigate the diagnostic value of lean test in Benign paroxysmal positional vertigo of posterior semicircular canal （PC-BPPV）. Methods We retrospectively included the clinical data of 220 patients with unilateral PC-BPPV who were admitted to the Department of Neurology，Jiaxing Second Hospital.All patients underwent lean test first，and then Dix-Hallpike test was performed to study the clinical characteristics of patients with vertigo induced by lean test. Results 133 （60.5%） patients induced PC-BPPV typical upbeat torsional nystagmus during the lean test. During the Dix-Hallpike test，the maximum slow phase angular velocity （SPV） of the induced upbeat nystagmus was higher than the SPV value induced by the lean test，and the latency and duration of the induced nystagmus were significantly different from the lean test （P 0.001）. Conclusion The lean test has a certain reference value for the diagnosis of PC-BPPV. It is effective for the diagnosis of PC-BPPV. The lean test induces a small nystagmus and mild autonomic symptoms compare to suspension position. In view of its simplicity，the lean test can be performed before the Dix-Hallpike operation.

Objective To explore the research hotspots and development trend on vascular dizziness/vertigo based on visual analysis. Methods The Web of Science Core Collection Database was searched for papers on vascular dizziness/vertigo from January 2008 to March 2023. The CiteSpace 6.2.R2 software was used for visual analysis of the literature. Results There were a total of 1298 papers，with an increasing number of published papers from January 2008 to March 2023. A total of 424 institutions from 83 countries/regions had published relevant papers. The United States ranked first in terms of the number of published papers（331 papers） and betweenness centrality（0.25）. Johns Hopkins University was the number one institution in terms of the number of published papers （56 papers），Newman-toker and David E were the most prolific authors. The most common keyword was ischemic stroke. According to keyword clustering，research in this field focused on early diagnosis of vascular dizziness/vertigo from risk factors and bedside examinations and infarction of the anterior inferior cerebellar artery supply area. In recent years，researchers had more interests in case reports，video electronystagmograms，and pathophysiological mechanisms in this field. Conclusion There are growing international studies on vascular dizziness/vertigo. Early diagnosis of vascular dizziness/vertigo through risk factors and bedside examinations in the emergency room is a research hotspot in this field. Researchers should focus on these topics in future studies.

Objective:To observe the effects of estradiol on expression of plasma membrane Ca 2+-ATPase isoform 2 in inner ear of rats. Methods:Twenty-five Three-months-old female Sprague-Dawley rats were randomly and equally divided into five groups by random number table mathod,with five rats in each group. Animals in Sham group were sham-operated while others were bilateral ovariactmized. One month after modeling, the OVX groups were supplemented with estradiol (E2 group), progesterone (P group), estradiol and progesterone (E2+P group)and vehicle sesame oil (Veh group), while the Sham operation group (Sham group) was supplemented with vehicle sesame oil.All rats were sacrificed and otocysts were obtained immediately. Enzyme-linked immunosorbent assay was used to detect the changes in serum estradiol and progesterone levels of each group of rats before operation, before treatment and before sacrifice. Western blot and quantitative real-time polymerase chain reaction were used to detect the expression of total PMCA2 protein and mRNA in the inner ear of each group.Results:There was no significant difference in serum estradiol and progesterone levels among the five groups before operated( P>0.05). Before treatment, the serum estradiol and progesterone levels of rats in each group were significantly lower than those in Sham group ( P<0.05). The serum estradiol level in E2 group and E2+P group was not significantly different from that in Sham group ( P>0.05), while the serum estradiol level in P group and Veh group was significantly different from that in Sham group ( P<0.05). The level of progesterone in P group and E2+P group was higher than that in Sham group ( P<0.05), while the level of progesterone in Veh group and E2 group was lower than that in Sham group ( P<0.05). Protein and mRNA expression of PMCA2 in P and Veh groups were significantly decreased compared with that of Sham group ( P<0.05) while the expression levels underwent no significantly change in E2 and E2+P groups ( P<0.05). Conclusion:The decrease of serum estradiol level can reduce the expression of otolith regulatory protein PMCA2 in rats, and then affect otolith metabolism, which may be an important link of estrogen affecting otolith metabolism.

Objective: To investigate the clinical value of serum adiponectin in patients with preeclampsia and its relationship with insulin resistance. Methods: From May 2017 to May 2018, 80 cases of preeclampsia diagnosed and treated in obstetrics department of the Second People′s Hospital of Liaocheng were selected in the research.According to the severity of the disease, they were divided into severe preeclampsia group and mild preeclampsia group, with 40 cases in each group, and 40 healthy pregnant women in the same period were selected as the control group.The serum adiponectin level and insulin resistance index (HOMA-IR) were detected and compared among the groups, and the correlation between them was analyzed. Results: The serum adiponectin and HOMA-IR in the severe preeclampsia group were (5.08 ±1.13)mg/L, (3.08 ±1.54), respectively, which in the mild preeclampsia group were (6.55±1.46)mg/L, (2.62±1.34), respectively, which in the control group were (11.67±3.53)mg/L, (1.13±0.53), respectively.there were statistically significant difference among the three groups (all P<0.05). The serum adiponectin level in the severe preeclampsia group was lower than that in the mild preeclampsia group and control group(P<0.05), and the HOMA-IR in the severe preeclampsia group was higher than that in the mild preeclampsia group and control group (P<0.05). The serum adiponectin level in the mild preeclampsia group was lower than that in the control group, and the HOMA-IR was higher than that in the control group, the differences were statistically significant (all P<0.05). The serum adiponectin level of preeclampsia patients with insulin resistance was (4.89±1.25)mg/L, which was significantly lower than that of non-insulin resistance patients[(6.78±1.75)mg/L], and the difference was statistically significant (t=6.254, P<0.05). Pearson correlation analysis showed that serum adiponectin level was negatively correlated with HOMA-IR (r=-0.617, P<0.05). Conclusion The serum adiponectin level and insulin resistance index of preeclampsia patients are significantly decreased, and there is a significant negative correlation between them.