[Objective] To explore the feasibility of using whole blood as a source of NK cells for allogeneic CAR NK cell therapy and activated NK cell reinfusion therapy, and initially construct a technical system for the separation and purification of NK cells from whole blood. [Methods] All peripheral blood mononuclear cells (PBMCs) were enriched from 400 mL of whole blood by manual separation and machine separation, respectively. The erythrocyte loss rate, PBMCs number, NK cell purity of the two methods were compared. NK cells were sorted from PBMCs by three separation and enrichment methods as immunomagnetic bead negative selection method, platelet lysate culture expansion and PERCOLL density gradient separation method, and the purity and yield of NK cells, the activity of NK cells and the tumor-killing ability of the three separation and enrichment methods were compared. [Results] The proportion of NK cells in the lymphocyte population was higher in the manual separation method than in the machine separation method[(13.16±5.16)% vs (8.56±3.92)%, P<0.05]; the number PBMCs was lower in the manual separation method than in the machine separation method[(4.09±1.80)×10⁸vs (6.49±2.16)×10⁸, P<0.05], and there was no difference in the red blood cell loss between the two methods (P>0.05). The purity of NK cells isolated and enriched from PBMCs by manual separation method using immunomagnetic was (96.77±2.31)%; the yield was (56.27±10.47)%; the inhibition of tumor proliferation was (38.67±14.05)%; and the tumor killing rate was (19.90±8.05)%. The purity of NK cells isolated and enriched from PBMCs by manual separation method using platelet lysis culture expansion method was the highest at day 7, which was (54.84±15.80)%; the cell expansion multiple could reach 16.92±6.28 at day 7; the in vitro tumor killing rate of NK cells was (15.83±5.5)%; the tumor inhibition rate was (44.33±13.5)%; and there was no difference in the toxicity and activity of NK cells between the two methods (P>0.05). The purity of NK cells isolated and enriched by PERCOLL density gradient separation method was (15.83±5.82)%, and the yield was (14±6.25)%, which was significantly lower than the other two methods. [Conclusion] PBMCs isolated from whole blood by manual separation and NK cells enriched by negative selection with immunomagnetic beads have the potential to provide NK cell materials for CAR-NK cell therapy, and NK cells enriched by platelet lysate-conditioned medium have the potential to provide NK cells for large-scale NK cell activation reinfusion therapy.

Neurofibromatosis type 1 (NF1) presents with a diverse range of symptoms that can affect the skin, bones, eyes, central nervous system, and other organs. This article reports the diagnosis and treatment process of a patient with NF1 complicated by bilateral severe-to-profound sensorineural hearing loss. Genetic testing revealed a heterozygous variant of NF1 NM_ 000267.3: c.4054_ 4058del(p.Ser1352LeufsTer20, supporting the diagnosis of NF1. After thorough evaluation, a right cochlear implantation was performed, yielding satisfactory postoperative results. This case suggests that NF1 patients may exhibit phenotypic heterogeneity and atypicality, providing a reference for clinicians in the diagnosis and treatment of such patients.

Objective:To explore the mediating effect of job resources between self-efficacy and job crafting behavior of Intensive Care Unit (ICU) nurses.Methods:From March to May 2022, 629 ICU nurses from Fuwai Hospital, Chinese Academy of Medical Sciences were selected by cluster sampling as the subjects. The nurses were investigated by using the General Information Questionnaire, Job Crafting Scale (JCS) , the Copenhagen Psychosocial Questionnaire (COPSOQ) , and the General Self-Efficacy Scale (GSES) . Pearson correlation was used to analyze the correlation between ICU nurses' job resources, self-efficacy and job crafting behavior. The mediating effect model was established by AMOS software, and the mediating effect was discussed by Bootstrap test. A total of 629 questionnaires were distributed, and 555 valid questionnaires were collected, and the effective response rate was 88.24% (555/629) .Results:Among 555 ICU nurses, the scores of JCS, job resources and GSES were (77.58±12.53) , (74.35±13.37) and (27.16±5.90) , respectively. Pearson correlation analysis showed that ICU nurses' job crafting behavior was positively correlated with self-efficacy and job resources ( P<0.01) . Bootstrap mediating effect analysis showed that there was a partial mediating effect of job resources between self-efficacy and job crafting behavior of ICU nurses [β= 0.128, 95% CI (0.086, 0.177) ] , accounting for 18.7% of the total effect. Conclusions:ICU nurses' job crafting behavior is at a moderate to upper level. Job resources play a mediating role between self-efficacy and job crafting behavior. Nursing managers should focus on the job crafting behavior of ICU nurses, build a supportive work environment, pay attention to their psychological health, give them correct guidance, and improve the self-efficacy of ICU nurses.

Objective:To compare the clinical effects of minimally invasive vitreous surgery with and without anti-vascular endothelial growth factor (VEGF) drugs for polypoid choroidal vascular disease (PCV) complicated with vitreous hemorrhage.Methods:A cohort study was performed.Thirty-six consecutive cases (36 eyes) with PCV combined with vitreous hemorrhage who underwent 25G minimally invasive vitreous surgery in Xuzhou First People's Hospital from June 2015 to June 2020 were enrolled.According to surgical methods, the patients were divided into pars plana vitrectomy (PPV) group (24 eyes) receiving vitrectomy only and PPV+ anti-VEGF group (12 eyes) receiving vitrectomy first and intravitreal injection of anti-VEGF drugs one week after the operation.All patients were followed up for at least 6 months.The best corrected visual acuity (BCVA) and central retinal thickness (CRT) of the two groups before treatment, 1 month after treatment and at the last follow-up were measured and compared.Postoperative complications such as recurrence of vitreous hemorrhage and macular scar formation were recorded.This study followed the Declaration of Helsinki and was reviewed and approved by the Medical Ethics Committee of Xuzhou First People's Hospital (No.xyyll[2021]014).Written informed consent was obtained from each patient before surgery.Results:Statistically significant differences were found in BCVA between the two groups before and after treatment ( Fgroup=8.552, P=0.006; Ftime=31.775, P<0.001).The BCVA of the two groups at 1 month after operation and at the last follow-up were significantly improved in comparison with before treatment, and the BCVA at the last follow-up was significantly better than that at 1 month after operation (all at P<0.05).One month after operation and at the last follow-up, the BCVA of PPV+ anti-VEGF group was better than that of PPV group, showing statistically significant differences (both at P<0.05).Statistically significant differences were found in CRT between the two groups before and after treatment ( Fgroup=4.797, P=0.041; Ftime=295.764, P<0.001).One month after operation and at the last follow-up, the CRT of both groups was significantly improved in comparison with before treatment, and the CRT was significantly better at the last follow-up than 1 month after operation (both at P<0.05).The postoperative 1-month CRT of PPV+ anti-VEGF treatment group was lower than that of PPV group, with statistically significant difference ( P<0.05).No statistically significant difference was found in CRT between the two groups at the last follow-up ( P>0.05).Elevated intraocular pressure occurred in 2 eyes and rhegmatogenous retinal detachment in 1 eye in PPV group, accounting for 8.33% and 4.17%, respectively.Cataract aggravated in 2 eyes in PPV+ anti-VEGF group.The incidence of vitreous rebleeding in PPV group and PPV+ anti-VEGF group was 16.67%(4/24) and 8.33%(1/12), respectively, with no significant difference ( P=0.646).The incidence of macular scarring in PPV group and PPV+ anti-VEGF group was 4.17%(1/24) and 33.3%(4/12), respectively, showing a statistically significant difference ( P=0.030). Conclusions:Minimally invasive vitreous surgery is a safe and effective way to treat PCV combined with vitreous hemorrhage.It can improve vision, reduce CRT, and the effect is gradually enhanced in the short term.Intravitreal injection of anti-VEGF drugs can enhance the postoperative effect of PPV and present better vision and anatomical structure of retina.

Objective To study the effects of prosthetic alignment on internal contact mechanical characteristics of intact knee joint for transfemoral amputees. Methods The gait experiment of transfemoral amputees was performed under different alignment conditions, and the differences of lower limb motion and ground reaction force (GRF) were analyzed and compared with those of the non-amputees. The three-dimensional （30） finite element model of knee joint was build and used to analyze the effect of alignments of socket adduction and abduction on internal contact mechanical characteristics between femur cartilage, tibia cartilage and meniscus. Results For knee joint of the non-amputees, contact force was mainly concentrated on the medial sides at the moment of the first GRF peak, while contact force was mainly concentrated on the lateral sides at the moment of the second GRF peak. However, for intact knee joint of the transfemoral amputees, contact force was mainly concentrated on the medial side at the moment of two GRF peaks. The stress of the medial meniscus, contact force and contact area between the medial meniscus and cartilage all obviously increased under the alignment of 6° socket adduction. Conclusions Compared with non-amputees, the incidence of knee osteoarthritis (OA) in amputees was higher, which was related to the long-term overload of the medial knee joint. The alignment of socket adduction may increase the risk of knee OA in the intact side of transfemoral amputees. In clinic, excessive adduction of the socket should be avoided during prosthetic alignment.

Exoskeleton nursing robot is a typical human-machine co-drive system. To full play the subjective control and action orientation of human, it is necessary to comprehensively analyze exoskeleton wearer's surface electromyography (EMG) in the process of moving patients, especially identifying the spatial distribution and internal relationship of the EMG information. Aiming at the location of electrodes and internal relation between EMG channels, the complex muscle system at the upper limb was abstracted as a muscle functional network. Firstly, the correlation characteristics were analyzed among EMG channels of the upper limb using the mutual information method, so that the muscle function network was established. Secondly, by calculating the characteristic index of network node, the features of muscle function network were analyzed for different movements. Finally, the node contraction method was applied to determine the key muscle group that reflected the intention of wearer's movement, and the characteristics of muscle function network were analyzed in each stage of moving patients. Experimental results showed that the location of the myoelectric collection could be determined quickly and efficiently, and also various stages of the moving process could effectively be distinguished using the muscle functional network with the key muscle groups. This study provides new ideas and methods to decode the relationship between neural controls of upper limb and physical motion.
Electromyography ; Exoskeleton Device ; Humans ; Muscle, Skeletal ; physiology ; Robotics ; Upper Extremity

Objective To investigate the activities of different soft and hard materials during right chewing masticatory muscles, describing the masticatory muscles of time domain and frequency domain features. Methods 11 experimental subjects who conform to the standards of measurement chew two materials of different soft and hard texture. Then record surface EMG of each bilateral temporalis anterior bundle, masseter, two bilateral anterior abdominal muscles, analysis to 5 kinds of characteristics of the study of EMG.Results When subjects chewing different soft and hard materials, al the EMG features in the bilateral anterior temporalis and masseter values had signifi cant difference (P < 0.05). The results in bilateral digastric anterior abdominal except zero crossing rate have signifi cant difference; median frequency on the right side of the masticatory muscle has decreased trend. ConclusionThe anterior temporalis and masseter EMG active more intense when chewing hard objects; the right side of the masticatory muscles have obvious fatigue trend after chewing hard masticatory 30 cycles.

The 3D gait analysis system (Vicon) was used for the gait analysis of health persons and amputees who wear lower limb prosthesis. The result indicated that gait analysis can supply changes of kinematics and biomechanics; which can be used to analyze the difference of joint movement between the health persons and amputees. The 3D gait analysis is feasible for amputees in rehabilitation.