BACKGROUND: Periventricular leukomalacia is a major syndrome of premature infant brain injury, which has been not prevented and cured yet. Theoretically, neural stem cells which were transplanted into white matter with an absence of oligodendroglial cells might be an ideal method to cure periventricular leukomalacia. OBJECTIVE: To prepare the multi-lineage potential of neural stem cells for the use of intraventricular transplantation. METHODS: Cerebral cortex was obtained from 12-14-day fetal rats and sectioned into 1.0-mm~3 sections. The single cell suspension was separated and purified. The neurospheres were incubated with DMEM/F12 culture medium containing fetal bovine serum to observe primary and passage culture of neural stem cells. The differentiation of neural stem cells was determined using immunohistochemical method. RESULTS AND CONCLUSION: The viability of cultured neural stem cells was (94.3±2.2)%. The neurosphere was formed at day 3 after primary culture. The proliferation of neurosphere slowed down after 10-passage culture, and some cells became old. All neurospheres were positively Nestin-staining, thus they were considered as neural stem cells. A further incubation of 4-passage neurospheres, immunohistochemical method indicated that the neurosphere was positively GFAP, β-tublin, and O4 staining, respectively. This suggested that cultured neural stem cells are able to self-renew, proliferate, and differentiate into neurons, astrocytes and oligodendroglial cells.

This study is to observe the effect of ilexonin A (IA) on the expression of basic fibroblast growth factor (bFGF) and growth associated protein-43 (GAP-43), and neurogenesis after cerebral ischemia-reperfusion in rats and explore its possible mechanism of protecting neuronal injury. Models of middle cerebral artery occlusion (MCAO) were established in SD rats. Before and after two hours ischemia-reperfusion, IA (20 and 40 mg x kg(-1)) was injected immediately and on 3, 7, 14, and 28 d once a day. The neurological severity was evaluated by neurological severity scores (NSS); neuronal injury in the boundary zone of the infarction area was evaluated by TUNEL and Niss1 staining. The expressions of bFGF and GAP-43 and neurogenesis were evaluated by Western blotting and 5-bromodeoxyuridine (Brdu) fluorescence staining, respectively. After treatment with IA, the NSS of treatment groups were lower than that of the models (3 and 7 d). The number of TUNEL positive neurons decreased and Nissl positive neurons increased at the same time (3 d). The expressions of bFGF and GAP-43 increased significantly in the boundary zone of the infarction area when compared to model group. Moreover, IA markedly enhanced the neurogenesis in the brain after ischemia-reperfusion, which revealed an increase of Brdu/NeuN positive cells in the boundary zone of the infarction area. The possible mechanism of protecting neuronal injury of IA may be related to inhibition on neuronal apoptosis, upregulation of bFGF and GAP-43, and neurogenesis in boundary zone of infarction after cerebral ischemia-reperfusion.

Objective To investigate the relationship between activation of gliacytes , mitogen-activated protein kinase (p38MAPK) and neuronal apoptosis after microinjecting aggregated Aβ25-35 into hippocampus.Methods The model was established by using stereotaxic technique to inject 10μg aggregated Aβ25-35 into dorsal hippocampus in rats .The rats were grouped as the control , vehicle and model groups .Immunohistochemistry and Western blotting were used for detection of activation of microglia(MG), atrocytes (AS) and expression of p-p38MAPK in the hippocampus.ELISA was used to evaluate the level of TNF-αand IL-1β.The survival neurons were observed by Nissl staining and the apoptotic neurons were identified by tunnel staining .Results Expression of ox-42, GFAP, p-p38MAPK were up-regulated in hippocampus, as well as TNF-α、IL-1β, which reached a highest value on the 7th day after injection of Aβ25-35.However, the number of neuron with Nissl positive decreased gradually , and the tunnel positive neurons increased highly and reached a peak value on the 7th day.There were significant differences between the control and vehicle group ( P <0.01). Conclusion Apoptosis of the neuron caused by Aβ25-35 injection may result from activation of gliacytes , p38 MAPK and increase of TNF-αand IL-1βlevel.

The probable mechanism of the reduction of rat cerebral ischemic-reperfusion injury by propyl gallate was studied. Intraluminal suture middle cerebral artery occlusion model of rat was employed. Propyl gallate was injected immediately after the ischemia was happened. The activity of NF-kappaB, and the expression of COX-2 and HSP70 on the peripheral ischemia were determined by Western blotting. The expression of TNF-alpha was determined by ELISA assay. RT-PCR and immunofluorescence staining were employed to detect the transcription and expression of TLR-4. Results showed that propyl gallate could inhibit the activity of NF-kappaB in the peripheral ischemia, and reduce the expression of COX-2 and TNF-alpha. As the upstream of NF-kappaB, the transcription and expression of TLR-4 decreased, as well as HSP70, the endogenic ligand of TLR-4. As an antioxidant, propyl gallate could reduce the cerebral ischemic-reperfusion injury through inhibiting the activity of NF-kappaB and decreasing the COX-2 and TNF-alpha in the peripheral ischemia. It also could influence HSP70 and TLR-4.

Objective To clarify the position,figure and connections with adjacencies in the pallial thickening(Pth),and provide essential parameters for its function study. Methods The coronal serial sections of 60?m thickness in gekko gecko brain were made by cryo-microtome,and Nissl staining was used.Pictures were taken in each coronal section containing the Pth and the size of Pth in each section was measured.One of them was chosen for the three-dimensional reconstruction.3D MAX was used as the tool software to rebuild the nucleus. Results 1.The Pth was located in the rostral part of the telencephalon,the lateral part of anterior dorsal ventricular ridge,the medial part of the lateral cortex and the ventral part of the dorsal cortex.The length of Pth from the rostral to the caudal end was(912.67?110.96)?m(n=10),the cubage of Pth was about(0.1430?0.0414)?m~3(n=10).2.The Pth could be divided into four segments,the anterior,the middle,the posterior and the terminal segments from the rostral to the caudal end.In the posterior segment,its dorsoventral axis was the longest,and could be divided into two parts: the dorsal and the ventral parts.The boundary of the two parts was clear.Conclusion The Pth is a long,narrow and flat structure;its rostrocaudal axis is longer than its dorsoventral axis,and its dorsal edge is smoother than its ventral edge.In the Pth,its caudal region is larger than its rostral region,and the posterior segment in the caudal region is divided into the dorsal and the ventral cell populations.

Objective To study the protective effect of curcumin on precurosor oligodendrocytes (preOLs) damage induced by hydrogen peroxide (H2O2),and explore its mechanism.Methods (1) In vitro primary culture ofpreOLs was performed; and 0 (normal controls),100,250 and 500 μmol/L H2O2 were added for 30 min; the apoptosis and death of preOLs were observed by Hoechst33342/PI double staining.(2) The preOLs were divided into normal control group,model group,and 5,10 and 20 μmol/L curcumin treatment groups; cells the later three groups were given 5,10 and 20 μmol/L curcumin for one h,and the later four groups were given 100 μmol/L H2O2 for 30 min; MTT assay was,then,employed to detect the cell viability.(3) The preOLs were divided into normal control group,model group,and 10 μmol/L curcumin treatment group; the apoptosis ofpreOLs was detected by Annexin V/FITC flow cytometry; Western blotting was used to detect the protein expressions of B cell lymphoma/leukemia-2 (Bcl2),Bcl-2 associated X protein (Bax) and cleaved caspase-3 and caspase-9; activities of total superoxide dismutase (T-SOD),glutathione peroxidase (GPx) and catalase (CAT) and malondialdehyde (MDA) were detected by spectrophotometry.Results (1) As compared with those in the normal control group,significantly decreased number of normal healthy cells and statistically increased apoptotic and necrotic cells in the 100,250 and 500 μmol/L H2O2 treatment groups were noted (P＜0.05); 100 μmol/L H2O2 treatment group had larger number of apoptotic cells than that of necrotic cells,while 250 and 500 μmol/L H2O2 treatment groups had larger number of necrotic cells than that of apoptotic cells.(2) Cells from 5,10 and 20 μmol/L curcumin treatment groups had significantly higher preOLs viability than those from model group,and 10 and 20 μmol/L curcumin treatment groups had significantly higher preOLs viability than those from 5 μmol/L curcumin treatment group (P＜0.05).(3) As compared with the model group,the 10 μmol/L curcumin treatment group had obviously decreased apoptotic and necrotic cells,increased activities of T-SOD,GPx and CAT,increased GSH level and decreased MDA concentration,up-regulated Bcl-2 expression,and inhibited Bax,caspase-3 and caspase-9 expressions,with significant differences (P＜0.05).Conclusion Curcumin has protective effect on preOLs against oxidative injury through effectively reducing lipid peroxidation,regulating Bcl-2/Bax expression and suppressing caspase-3 and caspase-9 activation.

The secondary damage after traumatic brain injury (TBI) involves a variety of pathological processes, and the inflammatory response in the nervous system is an important factor which affects nerve repair and regeneration. As the innate immune cell in the nervous system, microglia (MG) plays an important role in the entire neuroinflammatory environment by regulating the activation state of MG and changing the inflammatory response in the direction of promoting nerve repair and regeneration, which has great potential in treatment of TBI. This article reviews the inflammatory response of the nervous system after TBI and the reactivity of MG, as well as their significance in nerve repair and regeneration.

Objective:At present,the doctor-patient relationship is tense.The prevalence of negative emotions,such as depression and anxiety,among healthcare workers is increasing every year.Negative attitudes of medical workers toward mental problems may aggravate the doctor-patient conflict and psychological problems of medical workers.This study aims to explore the complex network relationships between outpatient medical workers'attitudes toward mental problems,doctor-patient relationships,and their depression/anxiety levels. Methods:A total of 578 outpatient medical staff from the Second Xiangya Hospital of Central South University(167 males,411 females)completed questionnaires on their attitudes toward mental problems,doctor-patient relationships,and depression/anxiety symptoms.Network analysis was conducted separately to construct the"attitude towards mental problems-doctor-patient relationship network"and"depression-anxiety related network". Results:The edge between"M15(insulting words)"and"D8(waste time)"showed the strongest strength in the"attitude towards mental problems-doctor-patient relationship network",and"M15(insulting words)"had the highest bridge strength in the network.For the analysis of emotional variables,"P1(anhedonia)"showed the most obvious association with"D10(communication difficulties)"in the doctor-patient relationship and"M2(poor quality of life)"in the psychiatric attitudes,and"P1(anhedonia)"was the key bridge symptom in the network. Conclusion:The"insulting words"may be an intervention target for medical workers'attitudes toward mental problems.The"anhedonia"in depression is the potential symptom that needs to be treated.Intervention targeting these variables may be beneficial to improve the mental health level of medical workers and the doctor-patient relationship.

Rutin,a flavonoid found in fruits and vegetables,is a potential anticancer compound with strong anti-cancer activity.Therefore,electrochemical sensor was developed for the detection of rutin.In this study,CoWO4 nanosheets were synthesized via a hydrothermal method,and porous carbon(PC)was prepared via high-temperature pyrolysis.Successful preparation of the materials was confirmed,and character-ization was performed by transmission electron microscopy,scanning electron microscopy,and X-ray photoelectron spectroscopy.A mixture of PC and CoWO4 nanosheets was used as an electrode modifier to fabricate the electrochemical sensor for the electrochemical determination of rutin.The 3D CoWO4 nanosheets exhibited high electrocatalytic activity and good stability.PC has a high surface-to-volume ratio and superior conductivity.Moreover,the hydrophobicity of PC allows large amounts of rutin to be adsorbed,thereby increasing the concentration of rutin at the electrode surface.Owing to the syn-ergistic effect of the 3D CoWO4 nanosheets and PC,the developed electrochemical sensor was employed to quantitively determine rutin with high stability and sensitivity.The sensor showed a good linear range(5-5000 ng/mL)with a detection limit of O.45 ng/mL.The developed sensor was successfully applied to the determination of rutin in crushed tablets and human serum samples.

Objective:To compare the therapeutic results between axis pedicle screwing assisted by intraoperative 3-D navigation and freehand axis pedicle screwing in the treatment of Hangman fracture.Methods:A retrospective analysis was performed of the 64 patients with Hangman fracture who had received posterior axis pedicle screwing at Department of Spinal Surgery, The Sixth Hospital of Ningbo from May 2014 to December 2019. According to the placement methods of axis pedicle screws, they were divided into a navigation group ( n=34, subjected to axis pedicle screwing assisted by intraoperative 3-D navigation) and a freehand group ( n=30, subjected to freehand axis pedicle screwing). Pedicle screw placement time, operation time, intraoperative bleeding, fluoroscopy time, hospital stay, total hospitalization cost and complications were recorded and compared between the 2 groups. The accuracy of axis pedicle screw placement was evaluated according to the postoperative cervical CT and screw grading criteria proposed by Park et al. At admission, 3 months postoperation, and the last follow-up, neurological function of the patients was evaluated by modified Japanese Orthopedic Association (mJOA) score, neck pain was evaluated by visual analogue scale (VAS), and C2/3 vertebral body angulation and C2 forward displacement were measured. The clinical efficacy was evaluated by Moon grading at the last follow-up. Results:The navigation group and the freehand group were comparable due to insignificant differences between them in the preoperative general data ( P>0.05). The accuracy of screw placement in the navigation group (98.2%, 54/55) was significantly higher than that in the freehand group (85.2%, 46/54) ( P<0.05). The screw placement time, operation time, fluoroscopy time and total hospitalization cost in the navigation group were significantly more than those in the freehand group ( P<0.05). Vertebral artery injury occurred in 3 cases in the freehand group. Screw loosening, screw breakage or rod breakage occurred in none of the patients after operation. There was no significant difference between the 2 groups in the intraoperative bleeding, hospital stay or follow-up time ( P>0.05). In both groups, the VAS score, mJOA score, C2/3 vertebral body angulation and C2 forward displacement were significantly improved at 3 months postoperation and the last follow-up compared with those at admission ( P<0.05), but there was no significant difference between the 2 groups in the contemporary comparisons ( P>0.05). At the last follow-up, Moon grading in the navigation group was significantly better than that in the freehand group ( P<0.05). Conclusion:In the treatment of Hangman fracture, compared with freehand screw placement, axis pedicle screwing assisted by intraoperative 3-D navigation can improve accuracy and safety of screw placement and reduce postoperative complications, leading to better clinical efficacy.