BACKGROUND:Neural stem cells (NSCs) hold self-renewal and multi-directional differentiation potential. NSCs differentiation into neurons in high proportion under induction conditions exhibits broad application prospect. OBJECTIVE:To explore the effect of basic fibroblast growth factor (bFGF)-chitosan carrier on the NSCs differentiation into neurons in vitro, and whether the differentiated neurons could form synaptic-like connection with myocytes. METHODS:After purification, the NSCs were co-cultured with chitosan, soluble bFGF or bFGF-chitosan carrier. After 7-day induction, the NSCs differentiation into neurons was observed by immunofluorescence staining of beta tubulin Ⅲ. The NSCs differentiation into cholinergic neurons was observed through double immunofluorescence staining of ChaT and beta tubulin Ⅲ. The synaptic-like connection between the neurons and myocytes was observed by triple staining of beta tubulin Ⅲ and MHC. RESULTS AND CONCLUSION:The percentage of differentiated neurons in the bFGF-chitosan carrier group was 74%, which was significantly higher than that in the other two groups. Additionally, the synaptic-like connection formed between the differentiated neurons and myocytes. To conclude, the bFGF-chitosan carrier promotes the NSCs differentiation into neurons to form synaptic-like connection with the co-cultured myocytes.

Objective To compare the application of CUBIC and iDISCO clearing methods in observing 3D imaging of spinal cord with immunofluorescent staining. Methods 1 mm thick spinal cord coronal sections were processed with CUBIC and iDISCO, respectively. The neurofilament (NF) protein was detected by immunofluorescent staining and then was observed by a laser confocal microscope. Results Compared with CUBIC, iDISCO had the advantages of shorter time, higher transparency (F=6.64, P<0.01), and deeper penetration (F=5117.55, P<0.01). Conclusion Immunofluorescent staining combined with iDISCO could completely observe the spinal axons with shorter time and better stain effect.

Objective To investigate the potential mechanism of basic fibroblast growth factor (bFGF)-chitosan carrier to induce neural stem cells to differentiate into neurons. Methods After purification, the neural stem cells were cocultured with chitosan, soluble bFGF and bFGF-chitosan carrier. Three hours, twenty-four hours, three days and seven days after induction, immunofluorescence staining of Nestin, beta tubulin III, microtubule-associated protein-2 (MAP2), and fibroblast growth factor receptor 1 (FGFR1) were used to observe the expres-sion of FGFR1;real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) and Western blotting were used to detect RNA and protein level changes after induction. Results Three hours after induction, there was no significant difference in the expression of FGFR1 among three groups. Twenty-four hours after induction, the expression level of FGFR1 was significantly higher in the bFGF-chitosan carrier group than in the chitosan group and the soluble bFGF group (P<0.001);three days and seven days after induction, the expression of FGFR1 decreased significantly in the chitosan group and soluble bFGF group (P<0.001), however, it was still higher in the bFGF-chitosan carrier group;moreover, the expression of genes associated with the pathway of extracellular regulated protein kinases/mitogen activated protein ki-nase (Erk/MAPK) was significantly higher in the bFGF-chitosan carrier group than in the chitosan group and soluble bFGF group (P<0.001). Conclusion bFGF-chitosan carrier might upregulate the expression of FGFR1, then activate Erk/MAPK signal pathways, and finally promote the differentiation of neural stem cells into neurons.

Objective To explore the problems and solving countermeasures in application of nursing vertical management,and provide references for nursing post management reform.Methods Through purposive sampling method,nursing administers,clinical nurses,heads of clinical departments and performance directors from five hospitals were interviewed by semi-structured interview method.Data were analyzed using 7-step Colaizzi's phenomenological analysis method.Results Four themes were formed about the problems in application of nursing vertical management,including:affecting department management and doctor-nurse collaboration,insufficient scientificity and fairness of performance evaluation,significant issues regarding post setting of senior nurses,competence and sense of belonging of temporarily deployed nurses.Four themes were formed about the solving countermeasures in application of nursing vertical management,including:improving participation of clinical departments in nursing management,making performance system to public and asking advices of nurses,providing career development for senior nurses,and conducting standardized training and establishing reserve nurses tank.Conclusion The main problems in application of nursing vertical management consisted of affecting department management and doctor-nurse collaboration,insufficient scientificity and fairness of performance evaluation,post setting of senior nurses,and competence and sense of belonging of temporarily deployed nurses.We suggested to better apply nursing vertical management by improving participation of clinical department in nursing management,making performance system to public and asking advices of nurses,providing career development for senior nurses,and conducting standardized training and establishing reserve nurses tank.

Objective To observe the effects of neurotrophin 3 (NT3)-chitosan on motor function, and proliferation and differentiation of the neural stem cells (NSCs) in the injury area and subventricular zone (SVZ) in rats with motor cortex injury. Methods Sixty-five Wistar rats were divided into control group (n=7), injury group (n=29) and NT3-chitosan group (n=29). The motor cortex was aspirated and re-moved as cerebral injury model. NT3-chitosan was immediately implanted into the injured area after operation, and the control group re-ceived no intervention. Pellet reaching test was performed to detect the recovery of the forelimb function, HE staining was used to observe the lesion cavity size, and immunofluorescence staining was used to observe the proliferation and differentiation of NSCs 3 days, 7 days, 14 days, 1 month, 2 months and 3 months after operation. Results The grasp success rate was higher (F>6.00, P≤0.05), and the lesion cavity size was significantly smaller (F>629.5, P<0.001) in the NT3-chitosan group than in the injury group. In the NSCs differentiation experi-ment, the number of BrdU cells at all the time points was significantly higher in the NT3-chitosan group than in the injury group (F>171.43, P<0.001). In the NSCs proliferation experiment, the number of BrdU positive cells was still significantly higher in the NT3-chitosan group than in the control group and in the injury group (F>155.06, P<0.001), the number of Dcx positive cells was significantly higher in the NT3-chitosan group than in the injury group (F=62.367, P<0.001), and the number of BrdU/Dcx positive cells was significantly higher in the NT3-chitosan group than in the control group (F=33.527, P<0.001). Conclusion NT3-chitosan could activate NSCs in the SVZ, and pro-mote endogenous neurogenesis and forelimb function recovery in rats after motor cortex injury.

Objective To investigate the effect of acupuncture at Baihui (DU20) and Zusanli (ST36) acupoints on the cognitive function and the oxidation protective mechanism of cortex in vascular cognitive impairment (VCI) rats, providing experimental evidence for the clinical treatment of VCI. Methods The VCI model was established by fluorescent microemboli injection through internal carotid artery. Randomly divided into the control group (n=12), the model group (n=12), the positive drug group (n=12), the acupuncture treatment group (n=12). Two weeks after modeling, the rats in the acupuncture treatment group were stimulated for 30 minutes with daily electroacupuncture at Baihui and Zusanli acupoints. The positive drug group was treated with donepezil hydrochloride 0.5206 mg/kg orally daily for 30 days. After the treatment, the water maze test was used to test the cognitive learning ability of rats. The content of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-Px) and hydrogen peroxide (H2O2) in the cortex of rats were detected by biochemical methods. Results Compared with the model group, the number of passing through the platform (7.5 ± 1.9, 6.8 ± 2.2 vs. 3.7 ± 1.0) of acupuncture treatment group and positive drug group were significantly higher (P<0.05), and the total distance in target quadrant (495.4 ± 89.4 cm, 487.6 ± 96.2 cm vs. 341.4 ± 67.3 cm) were significantly longer (P<0.05). In acupuncture treatment group and the positive drug group, the activity of SOD in the cortex (17.3 ± 3.3 U/mg, 15.1 ± 2.5 U/mg vs. 9.7 ± 4.9 U/mg) was significantly higher (P<0.05), but the MDA (9.1 ± 2.2 μmol/L, 8.4 ± 3.7 μmol/L vs. 15.2 ± 4.4μmol/L), and H2O2 (85.2 ± 16.2 μmol/L, 82.1 ± 13.2 μmol/L vs. 114.7 ± 24.8 μmol/L) were significantly lower (P<0.05). In acupuncture group, the activity of GSH-Px (14.5 ± 3.7 U/mg vs. 9.0 ± 2.5 U/mg) was significantly higher than that in control group (P<0.05). There was no significant difference between acupuncture group and positive drug (P>0.05). Conclusions Acupuncture at Baihui and Zusanli acupoints can improve the behavior scores of rats with VCI, increase the contents of total SOD and GSH-Px,decrease the content of MDA and H2O2 in brain,and enhance antioxidant effects, while inhibiting peroxidation, improve free radical metabolism.

Objective: To investigate the value of Multislice computed tomography volume rendering(VR) technique and 3D printing technique in auricular reconstruction. Methods: Six patients were enrolled for auricular reconstruction with costal cartilage, including 5 congenital microtia patients and 1 traumatic auricular defect patient. We harvest the three-dimensional reconstructive data of the contralateral sixth, seventh, eighth and ninth costal cartilage with VR technique. Three-dimensional solid models were 3D printed with nylon material according to the data exported in STL format. Preoperative simulation was performed on the models, accordingly, we determined the strategies of costal cartilage harvest and framework fabrication, and operations were performed based on the pre-designed plan. Results: In all 6 patients, the actual costal cartilage harvest and framework fabrication process was consistent with the preoperative design and simulation results, and more scientific than before. The shapes of reconstructed ears were vivid and natural. No complications such as infection, absorption, distortion and chest deformity happened. Conclusions Through costal cartilage VR and 3D printing technique, we could make more reasonable preoperative design and simulation. The results can be improved with reduced injury, while avoiding the risks of thoracic deformity.

Objective:To observe the morphological characteristics of high myopia (HM) paravalvular abnormalities (PVA), and the correlation between different manifestations of PVA and myopic traction maculopathy (MTM) was analyzed.Methods:A cross-sectional clinical study. A total of 42 middle-aged and elderly patients with HM and PVA diagnosed by ophthalmology examination in Department of Ophthalmology, The Second Hospital of Hebei Medical University from June to December 2021 were included in the study. There were 24 eyes in 16 males and 48 eyes in 26 females. Age was (56.71±8.10) years old. Diopter was (-13.05±3.10) D. Axial length (AL) was (28.22±1.04) mm. According to the characteristics of ultra-wide-angle optical coherence tomography images, PVA morphology was divided into paravascular microfolds (PM), paravascular cysts (PC) and paravascular lamellar holes (PLH). MTM was divided into T0-T5 grades, of which MTM≥T3 was defined as severe MTM. The state of vitreoretinal junction was observed and the state of posterior vitreous detachment (PVD) was recorded, which divided into complete PVD and partial PVD. Partial PVD was divided into macular fovea adhesions and paravascular adhesions according to the vitreoretinal adhesions. Posterior scleral staphyloma (PS) was divided into 6 types by ultra-wide-angle fundus photography. Logistic regression model was used to analyze the factors related to MTM.Results:In 72 eyes, PM, PC and PLH were 72 (100.0%, 72/72), 62 (86.1%, 62/72) and 29 (40.3%, 29/72) eyes, respectively. Among them, there were 10 (13.9%, 10/72) eyes with PM alone, 33 (45.8%, 33/72) eyes with PM and PC, and 29 (40.3%, 29/72) eyes with PM, PC and PLH, respectively. There were 42 eyes with partial PVD (58.3%, 42/72), among which the macular fovea and paravascular adhesion were 22 (52.4%, 22/42) and 24 (57.1%, 24/42) eyes, respectively. PS was present in 50 eyes (69.4%, 50/72), among which 27 (54.0%, 27/50), 21 (42.0%, 21/50), 1 (2.0%, 1/50), and 1 (2.0%, 1/50) eyes were types Ⅰ to Ⅳ, respectively. Multivariate logistic regression analysis showed that AL[odds ratio ( OR)=16.139, 95% confidence interval ( CI) 4.062-64.120, P<0.001], PS ( OR=4.212, 95% CI 1.234-14.378, P=0.022), paravascular vitreoretinal adhesion (OR=3.478, 95% CI 11.124, P=0.036) were risk factors for PM, PC and PLH. MTM was present in 58 eyes (80.6%, 58/72), among which T1 was the most common type in 19 eyes (26.4%, 19/72). Univariate logistic regression analysis showed that the occurrence of MTM was significantly correlated to PS ( OR=4.190, 95% CI 1.240-14.157, P=0.021), coexistence of PM, PC and PLH ( OR=11.323, 95% CI 1.389-92.311, P=0.023), and PS were significantly correlated. There was no correlation with PVD ( OR=1.889, 95% CI 0.580-6.150, P=0.291) or PS ( OR=2.778, 1.786; 95% CI 0.700-11.023; 0.445-7.167; P=0.146, 0.413). There was significant difference in the incidence of severe MTM between PM alone, PM combined with PC and coexistence of PM, PC and PLH ( χ2=20.943, P<0.001). Conclusions:PM is the most common and earliest manifestation of PVA in middle-aged and elderly HM patients. AL, PS and paravascular vitreoretinal adhesion are risk factors for PM, PC and PLH. The coexistence of three PVA forms may be a marker of severe MTM.

Objective: To explore the effectiveness and safety of ultrasound-guided radiofrequency ablation (RFA) in treatment of thyroid benign nodules. Methods: We analyze 573 patients with thyroid benign nodules from June 2014 to September 2017 treated by RFA at Department Ⅱof Thyroid Surgery, the First Affiliated Hospital of Zhengzhou University. Among these patients, there were 75 males and 498 females, with a median age of 45 years old. All patients were diagnosed as thyroid benign nodules by ultrasound-guided fine needle aspiration biopsy before RFA. A total of 750 benign tumors were treated. To evaluate the thyroid function of the patients before RFA and 3 months after it, and to observe the changes of thyroid benign nodules by ultrasound at 3, 6, 12 months after RFA. The paired t-test was used to compare the measurement data with normal distribution, and Wilcoxon's signed rank test was used to compare the measurement data with non-normal distribution. To calculate the volume change and reduction rate of thyroid benign nodules. Results: RFA was successfully completed in all patients, the volume reduction rate was 67%(48%, 83%) in the 3rd month after RFA, in the 6th month was 81%(67%, 91%), in the 12th month was 89%(80%, 95%). Eighteen patients felt pain during RFA, but the pain was alleviated after stopping ablation. Three patients′ tone decreased, but recovered in a week. Hoarseness occurred in 6 patients and recovered in 3 months. Three patients had neck hemorrhage, which was managed with simple compression of the neck. Conclusions RFA is an effective and safe treatment for thyroid benign nodules and has obvious advantages such as less invasiveness, having no influence in thyroid functions. It is clinically prospective for application.

Objective:To investigate the effects of exosomes of human nucleus pulposus cells (NPCs) on the differentiation of urine derived stem cells (USCs) into nucleus pulposus-like cells.Methods:USCs and NPCs were isolated and cultured in vitro. The exosomes of NPCs were extracted and detected by Western-blot. USCs cytoplasm was transfected with GFP lentivirus, while nucleus was transfected with DAPI dye. The NPCs exosomes were transfected with PKH26 dye. After co-incubation for 12 h, USCs and NPCs exosomes were observed by macroscopy. USCs differentiation was induced by NPCs exosomes and non-contact co-culture methods. The relative expression of marker gene mRNA of nucleus pulposus cells in each group and the absorbance at 450 nm wavelength were detected.Results:The isolated USCs had the ability to differentiate into osteocytes, adipocytes and chondrocytes with high expression of marker CD29 (99.57%), CD44 (97.46%) and CD73 (97.71%) and with low expression of negative proteins CD31 (0.59%) and CD45 (0.19%). The isolated NPCs highly expressed nuclear pulposus cell marker COL2A1, ACAN and SOX-9. The exosomes extracted from NPCs showed high expression of exosome marker CD63, CD81 and Tsg101. After 12 h co-incubation, NPCs exosomes fused with USCs membrane and appeared in the cytoplasm of USCs. At 3, 5 and 7 days of co-culture, the absorbance value of USCs cells in exosome group (0.44±0.004, 0.76±0.004, 0.82±0.006) was higher than that in co-culture group (0.39±0.022, 0.63±0.035, 0.69±0.012) ( P<0.05). The mRNA relative expression of USCs nucleus pulposus marker genes ACAN (1.80±0.31, 3.50±0.21, 5.35±0.31, 7.46±0.12), COL2A1 (1.43±0.15, 4.33±0.23, 6.89±0.22, 8.11±0.31), SOX-9 (2.21±0.13, 3.13±0.11, 3.96± 0.14, 4.52±0.26) and HIF-1α (1.45±0.16, 2.14±0.21, 4.31±0.41, 4.01±0.25) in exosomes group were significantly higher than those in the control group ( P<0.05) at the 3rd, 7th, 14th and 21st days. The mRNA relative expression of USCs nucleus pulposus marker genes ACAN (5.69±0.21, 6.69±0.13), COL2A1 (6.33±0.17, 7.89±0.15), SOX-9 (4.19±0.29, 4.38±0.12), HIF-1α (4.49±0.32, 4.96±0.26) in exosomes group were significantly higher than those ACAN (3.69±0.35, 5.13±0.23), COL2A1 (3.40±0.16, 6.79±0.19), SOX-9 (2.26±0.32, 3.69±0.26), HIF-1α (2.39±0.11, 3.96±0.13) in non-contact co-culture group ( P<0.05) at the 14th and 21st days. Conclusion:Human nucleus pulposus exosomes could induce differentiation of human USCs into nucleus pulposus-like cells in vitro. Compared with non-contact co-culture, exosomes have higher induction efficiency and can better maintain the proliferation activity of nucleus pulposus-like cells