Objective: To investigate the effects of clinical routine X-ray irradiation dose (average irradiation dose: 29.7±0.54 Gy) on the function, apoptosis, activation state and mitochondrial function of platelets during in vitro storage, so as to provide experimental evidence for optimizing platelet irradiation strategies. Methods: A paired experimental design was adopted. Platelets were collected from 12 healthy donors, and each sample was equally divided into the irradiated group and the control group (non-irradiated). All samples were stored for 5 days under standard platelet preservation conditions (22±2℃, continuous oscillation). Flow cytometry was used to detect platelet count, apoptosis rate (Annexin V+ positive rate), activation markers (CD62P, PAC-1, CD42b) and reactive oxygen species (ROS) level. Meanwhile, mitochondrial-specific probes were used to evaluate changes in mitochondrial count, membrane potential and adenosine triphosphate (ATP) content. Additionally, transmission electron microscopy (TEM) was employed to observe the ultrastructure of platelets, with a focus on mitochondrial morphology, platelet membrane integrity and granule distribution. Results: Within 5 days of storage, the platelet count was (841±89.16)×10 /L in the irradiated group and (824.5±92.88)×10 /L in the control group, with no statistically significant difference between the two groups (P=0.54). The apoptosis rate was (4.94±1.39) % in the irradiated group and (5.50±0.83) % in the control group, showing no significant difference (P=0.31). For activation indicators, the CD62P expression rate was (24.32±7.57) % in the irradiated group versus (25.21±8.13) % in the control group (P=0.43). The PAC-1 positive rates were (12.15±4.43) % and (11.75±3.40) % in the irradiated group and control group, respectively (P=0.44). The CD42b expression rates were (12.14±4.43) % and (11.75±3.4) % in the two groups, respectively (P=0.47). The ROS levels were (31.98±8.1) % and (30.64±5.89) % in the two groups, respectively (P=0.45). No significant differences were found in the above indicators. For mitochondrial function indicators, the mitochondrial count was (55.88±11.49) % in the irradiated group and (53.5±7.24) % in the control group (P=0.57). The ATP contents were (42.45±5.29) % and (41.58±9.50) % in the irradiated group and control group, respectively (P=0.77). The relative membrane potential values were (59.53±10.89) % and (57.49±6.54) % in the two groups, respectively (P=0.47). No significant difference were observed on the mitochondrial function-related indicators. TEM further confirmed that the ultrastructure of platelets in the irradiation group was intact, the mitochondrial morphology was normal, and no pathological changes such as swelling or vacuolization were observed. Conclusion: This study evaluated the impact of conventional-dose X-ray irradiation on platelet storage quality, confirming that this dose does not significant impair platelet count, apoptosis rate, activation status, or mitochondrial function. This finding provides important experimental evidence for the clinical promotion of X-ray irradiation technology and suggests its potential as a safe alternative to γ irradiation. Future studies could further expand the sample size and extend the observation period to verify the effects of X-ray irradiation on long-term platelet storage and post-transfusion in vivo survival rate.

Objective:To explore the effect of combining intermittent θ pulse stimulation (iTBS) of the cerebellum with lower extremity exoskeleton robot support on the balance and walking function of stroke survivors.Methods:Seventy-five stroke survivors complicated with lower extremity dysfunction were divided into an iTBS group, an exoskeleton group and a combined group, each of 25, according to a random number table. In addition to conventional rehabilitation training, the iTBS group was given cerebellar iTBS combined with traditional walking training, the exoskeleton group received sham cerebellar iTBS combined with walking training assisted by a lower extremity exoskeleton robot. The combined group received both therapies. The schedule was once a day, 5 days a week for 3 weeks. Before and after the treatment, the 10-metre walking test (10MWT), the Berg Balance Scale (BBS) and the Fugl-Meyer lower extremity assessment (FMA-LE) were used to evaluate the subjects′ walking ability, balance and lower extremity motor ability. Gait and neuro-electrophysiological tests were also conducted in all three groups.Results:After the treatment, a significant improvement was observed in the 10MWT times, BBS scores, FMA-LE scores, stride frequency and stride speed of all three groups compared with before the treatment. On average, the results of the exoskeleton and combined groups were significantly better than those of the iTBS group, and those of the combined group were significantly better than among the exoskeleton group. Almost everyone′s MEP latency and amplitude had improved significantly compared with before the treatment, but the improvements in the exoskeleton group tended to be superior to those in the iTBS group ( P≤0.05). The latency in the combined group averaged (21.25±1.70)ms, and the amplitude averaged (184.17±6.54)μV, both significantly better than the exoskeleton group′s averages. Conclusions:Cerebellum iTBS combined with lower extremity exoskeleton walker training can significantly improve the motor functioning, balance and walking ability of stroke survivors.

Objective:To explore the effect of combining repeated peripheral (rPMS) and central transcranial magnetic stimulation (rTMS) in treating upper limb motor dysfunction after a stroke.Methods:Seventy-eight patients with upper limb motor dysfunction after a stroke were randomly divided into a control group, an rTMS group and a combined magnetic stimulation group, each of 26. All three groups underwent routine rehabilitation, while the rTMS group was repeatedly given low frequency transcranial magnetic stimulation of the M1 region on the unaffected side, and the combined group also received repeated peripheral magnetic stimulation at Erb′s point on the affected upper limb. There was one treatment session a day, 5 days a week for 3 weeks. Before and after the treatment, everyone′s upper limb motor function was quantified using the Fugl-Meyer upper extremity assessment (FMA-UE) and the Wolf motor function test (WMFT). Skill in the activities of daily living was quantified in terms of a Barthel index (BI). Motor recovery of the upper limbs and hands was assessed using Brunnstrom staging. The latency and amplitude of the motor evoked potentials (MEPs) in the subjects′ affected abductor pollicis brevis muscles were also recorded before and after the treatment. Pearson correlation coefficients quantified the correlation between the changes in FMA-UE scores and MEP amplitudes before and after the treatment in the three groups.Results:There were no significant differences among the three groups before the treatment. Afterward, however, the average FMA-UE, WMFT and BI scores, as well as the upper limb and hand Brunnstrom stages and the average MEP latencies and amplitudes of all the three groups had improved significantly. The combined group′s average results were then significantly better than the other two groups′ averages, except for the upper limb Brunnstrom stages. The increases in MEP amplitude were positively correlated with the increases in FMA-UE scores among the rTMS and the combined group, but there was no significant correlation between them in the control group.Conclusions:The combined application of rPMS and contralateral low frequency rTMS can effectively relieve motor dysfunction in the upper limbs in the early stages after a stroke.

Objective To observe resting-state functional connectivity(rs-FC)changes of mirror neuron system(MNS)brain regions in male nicotine addicts,and to explore their correlations with the degree of addiction.Methods Twenty-five male nicotine addicts(addiction group)and 27 healthy males(control group)were prospectively recruited,and resting-statefunctional MR scanning was performed.The general information and rs-FC values were compared between groups,and correlations of rs-FC values with Fagerstr?m test for nicotine dependence(FTND)and heaviness of smoking index(HSI)scale scores in addiction group were analyzed.Results No significant difference of age nor education level was found between groups(both P＞0.05).Compared to control group,rs-FC values between left inferior frontal gyrus(IFG.L)and right middle temporal gyrus(MTG.R)as well as right middle occipital gyrus(MOG.R)increased(GRF corrected,voxel-level P＜0.005,cluster-level P＜0.05),while between right fusiform gyrus(FFG.R)and right dorsal cingulate gyrus(DCG.R),right supplementary motor area(SMA.R),left medial superior frontal gyrus(SFGmed.L)and left supplementary motor area(SMA.L)decreased(GRF corrected,voxel-level P＜0.005,cluster-level P＜0.05).In addiction group,rs-FC value between IFG.L and MTG.R was negatively correlated with HSI scale score(r=-0.457,P=0.022),while that between IFG.L and MOG.R was negatively correlated with HSI scale and FTND scores(r=-0.440,-0.458,P=0.028,0.021).Conclusion There were specific changes of rs-FC patterns of MNS brain regions in male nicotine addicts,which were correlated with the degree of addiction.

Objective:To observe the effect of combining transcutaneous stimulation of the auricular vagus nerve (taVNS) with modified constraint-induced movement therapy (mCIMT) on the upper limb function of hemiplegic stroke survivors.Methods:Seventy-one hemiplegic stroke survivors were randomly divided into a taVNS group of 24, an mCIMT group of 23 and a combined group of 24. In addition to conventional rehabilitation therapy, the taVNS group received taVNS therapy, the mCIMT group received mCIMT, while the combined group received both for 30 minutes a day, 5 days a week for 4 weeks. Before and after the treatment, everyone′s upper limb function was evaluated using the Fugl-Meyer upper extremity assessment (FMA-UE) and the Action Research arm test (ARAT). Ability in the activities of daily living (ADL) was quantified using the modified Barthel index (MBI). The latency and amplitude of motor evoked potentials (MEPs) were also measured.Results:After the treatment, significant improvement was observed in the average FMA-UE, ARAT and MBI scores. MEP latency and amplitude had also improved. The averages in the mCIMT group were then significantly better than among the taVNS group.Conclusions:taVNS combined with mCIMT can significantly improve the upper limb function and ADL functioning of stroke survivors with hemiplegia. It can also promote the recovery of central nervous system function.

Objective To observe resting-state functional connectivity(rs-FC)changes of mirror neuron system(MNS)brain regions in male nicotine addicts,and to explore their correlations with the degree of addiction.Methods Twenty-five male nicotine addicts(addiction group)and 27 healthy males(control group)were prospectively recruited,and resting-statefunctional MR scanning was performed.The general information and rs-FC values were compared between groups,and correlations of rs-FC values with Fagerstr?m test for nicotine dependence(FTND)and heaviness of smoking index(HSI)scale scores in addiction group were analyzed.Results No significant difference of age nor education level was found between groups(both P＞0.05).Compared to control group,rs-FC values between left inferior frontal gyrus(IFG.L)and right middle temporal gyrus(MTG.R)as well as right middle occipital gyrus(MOG.R)increased(GRF corrected,voxel-level P＜0.005,cluster-level P＜0.05),while between right fusiform gyrus(FFG.R)and right dorsal cingulate gyrus(DCG.R),right supplementary motor area(SMA.R),left medial superior frontal gyrus(SFGmed.L)and left supplementary motor area(SMA.L)decreased(GRF corrected,voxel-level P＜0.005,cluster-level P＜0.05).In addiction group,rs-FC value between IFG.L and MTG.R was negatively correlated with HSI scale score(r=-0.457,P=0.022),while that between IFG.L and MOG.R was negatively correlated with HSI scale and FTND scores(r=-0.440,-0.458,P=0.028,0.021).Conclusion There were specific changes of rs-FC patterns of MNS brain regions in male nicotine addicts,which were correlated with the degree of addiction.

Objective:To observe the effect of combining transcutaneous stimulation of the auricular vagus nerve (taVNS) with modified constraint-induced movement therapy (mCIMT) on the upper limb function of hemiplegic stroke survivors.Methods:Seventy-one hemiplegic stroke survivors were randomly divided into a taVNS group of 24, an mCIMT group of 23 and a combined group of 24. In addition to conventional rehabilitation therapy, the taVNS group received taVNS therapy, the mCIMT group received mCIMT, while the combined group received both for 30 minutes a day, 5 days a week for 4 weeks. Before and after the treatment, everyone′s upper limb function was evaluated using the Fugl-Meyer upper extremity assessment (FMA-UE) and the Action Research arm test (ARAT). Ability in the activities of daily living (ADL) was quantified using the modified Barthel index (MBI). The latency and amplitude of motor evoked potentials (MEPs) were also measured.Results:After the treatment, significant improvement was observed in the average FMA-UE, ARAT and MBI scores. MEP latency and amplitude had also improved. The averages in the mCIMT group were then significantly better than among the taVNS group.Conclusions:taVNS combined with mCIMT can significantly improve the upper limb function and ADL functioning of stroke survivors with hemiplegia. It can also promote the recovery of central nervous system function.

Objective:To explore the effect of combining intermittent θ pulse stimulation (iTBS) of the cerebellum with lower extremity exoskeleton robot support on the balance and walking function of stroke survivors.Methods:Seventy-five stroke survivors complicated with lower extremity dysfunction were divided into an iTBS group, an exoskeleton group and a combined group, each of 25, according to a random number table. In addition to conventional rehabilitation training, the iTBS group was given cerebellar iTBS combined with traditional walking training, the exoskeleton group received sham cerebellar iTBS combined with walking training assisted by a lower extremity exoskeleton robot. The combined group received both therapies. The schedule was once a day, 5 days a week for 3 weeks. Before and after the treatment, the 10-metre walking test (10MWT), the Berg Balance Scale (BBS) and the Fugl-Meyer lower extremity assessment (FMA-LE) were used to evaluate the subjects′ walking ability, balance and lower extremity motor ability. Gait and neuro-electrophysiological tests were also conducted in all three groups.Results:After the treatment, a significant improvement was observed in the 10MWT times, BBS scores, FMA-LE scores, stride frequency and stride speed of all three groups compared with before the treatment. On average, the results of the exoskeleton and combined groups were significantly better than those of the iTBS group, and those of the combined group were significantly better than among the exoskeleton group. Almost everyone′s MEP latency and amplitude had improved significantly compared with before the treatment, but the improvements in the exoskeleton group tended to be superior to those in the iTBS group ( P≤0.05). The latency in the combined group averaged (21.25±1.70)ms, and the amplitude averaged (184.17±6.54)μV, both significantly better than the exoskeleton group′s averages. Conclusions:Cerebellum iTBS combined with lower extremity exoskeleton walker training can significantly improve the motor functioning, balance and walking ability of stroke survivors.

Objective:To explore the effect of combining repeated peripheral (rPMS) and central transcranial magnetic stimulation (rTMS) in treating upper limb motor dysfunction after a stroke.Methods:Seventy-eight patients with upper limb motor dysfunction after a stroke were randomly divided into a control group, an rTMS group and a combined magnetic stimulation group, each of 26. All three groups underwent routine rehabilitation, while the rTMS group was repeatedly given low frequency transcranial magnetic stimulation of the M1 region on the unaffected side, and the combined group also received repeated peripheral magnetic stimulation at Erb′s point on the affected upper limb. There was one treatment session a day, 5 days a week for 3 weeks. Before and after the treatment, everyone′s upper limb motor function was quantified using the Fugl-Meyer upper extremity assessment (FMA-UE) and the Wolf motor function test (WMFT). Skill in the activities of daily living was quantified in terms of a Barthel index (BI). Motor recovery of the upper limbs and hands was assessed using Brunnstrom staging. The latency and amplitude of the motor evoked potentials (MEPs) in the subjects′ affected abductor pollicis brevis muscles were also recorded before and after the treatment. Pearson correlation coefficients quantified the correlation between the changes in FMA-UE scores and MEP amplitudes before and after the treatment in the three groups.Results:There were no significant differences among the three groups before the treatment. Afterward, however, the average FMA-UE, WMFT and BI scores, as well as the upper limb and hand Brunnstrom stages and the average MEP latencies and amplitudes of all the three groups had improved significantly. The combined group′s average results were then significantly better than the other two groups′ averages, except for the upper limb Brunnstrom stages. The increases in MEP amplitude were positively correlated with the increases in FMA-UE scores among the rTMS and the combined group, but there was no significant correlation between them in the control group.Conclusions:The combined application of rPMS and contralateral low frequency rTMS can effectively relieve motor dysfunction in the upper limbs in the early stages after a stroke.

Objective:To explore the changes in the functional connectivity (FC) of hippocampal subregions in nicotine addicts after repetitive transcranial magnetic stimulation (rTMS) using resting-state FC.Methods:This study was a cross-sectional study. The clinical and imaging data from 20 male nicotine addicts at Zhejiang Hospital between 2022 and 2024 were analyzed prospectively. All participants received rTMS treatment and were assessed with nicotine-related clinical scales and resting-state FC analysis before and after treatment. The clinical scale evaluations included the Fagerstr?m test for nicotine dependence (FTND), smoking severity index, Minnesota nicotine withdrawal scale (MNWS), short tobacco craving questionnaire (sTCQ), and visual analog scale (VAS). Paired t-tests and Wilcoxon signed-rank tests were used to compare the differences in clinical scale scores before and after treatment. Ten subregions of the bilateral hippocampus (including the hippocampus, dentate gyrus, entorhinal cortex, hippocampus-amygdala transition area, and subiculum) were used as seed points, and paired t-tests were conducted to compare the FC differences in these subregions before and after treatment. Pearson and Spearman correlation analyses were used to assess the correlation between changes in resting-state FC in the rTMS group and clinical scale scores. Results:Compared to pre-treatment, the scores on the FTND, smoking severity index, MNWS, sTCQ, and VAS all significantly decreased after rTMS treatment in nicotine addicts (all P<0.05). Compared to pre-treatment, post-treatment FC was reduced between the left dentate gyrus subregion and the bilateral supplementary motor area and left middle cingulate gyrus, while it increased between the left entorhinal cortex subregion and the right middle and superior temporal gyri, and between the left hippocampus-amygdala transition area subregion and the bilateral calcarine cortex and cuneus (Gaussian random field correction, voxel-level P<0.01, cluster-level P<0.05). Negative correlations were observed between the FC difference in the left hippocampus-amygdala transition area subregion and the right calcarine cortex and the difference in sTCQ-impulse score before and after treatment ( r=-0.447, P=0.048). Negative correlations were observed between the FC difference in the left hippocampus-amygdala transition area subregion and the right cuneus and the difference in the sTCQ-expectation score ( r=-0.559, P=0.010). Negative correlations were observed between the FC difference in the left hippocampus-amygdala transition area subregion and the left calcarine cortex and the differences in sTCQ-emotion and sTCQ-expectation scores ( r=-0.516, P=0.020; r=-0.466, P=0.038, respectively). Negative correlations were observed between the FC difference in the left hippocampus-amygdala transition area subregion and the left cuneus and the differences in sTCQ-emotion and sTCQ-expectation scores ( r=-0.459, P=0.042; r=-0.501, P=0.024, respectively). Conclusion:Changes in FC in certain hippocampal regions are observed in nicotine addicts following rTMS treatment, suggesting that hippocampal subregions may serve as potential biomarkers for nicotine addiction withdrawal to some extent.