BackgroundMild cognitive impairment (MCI) is a precursor state of Alzheimer’s disease (AD) and has attracted attention, but why amnestic mild cognitive impairment (aMCI) is more likely to progress to AD than non-amnestic mild cognitive impairment (naMCI) is unclear. The present study of aMCI compares differences in intra- and inter-network functional connectivity (FC) across multiple networks in naMCI and further correlates FC with cognitive assessment scores to assess their ability to predict AD progression.MethodsResting-state functional magnetic resonance imaging (rs-fMRI) was performed in 30 naMCI and 40 aMCI cases, and 12 resting-state networks (RSNs) were identified by independent component analysis (ICA). Two-sample t-tests were performed to detect intra-network FC differences, and functional network connectivity (FNC) was calculated to compare inter-network FC differences. Subsequently, Pearson or Spearman correlation analyses were used to explore the correlation between altered FC and cognitive assessment scores.ResultsThe aMCI compared to the naMCI differed within the (Default mode network) DMN, (Dorsal attention network) DAN, (Sensorimotor system) SMN, and (Salience network) SN networks (corrected for FWEc, P< 0.05), and inter-network differences in DAN-DMN, DMN-SN, SN-SMN (corrected for FWEc, P<0.05).ConclusionaMCI contrasts naMCI with widespread intra- and inter-static FNC differences, mainly involving the DMN, DAN, SMN, and SN. these network interactions provide a powerful method for assessing and predicting why aMCI is more likely to progress to AD, and contribute to our understanding of the neurological mechanisms underlying the pathological process of AD.

ObjectiveThis study aims to utilize multimodal neuroimaging techniques to simultaneously analyze global topological properties of white matter structural networks and resting-state functional networks in aMCI patients, comparing them with healthy controls. By conducting independent and integrative analyses of topological impairments in both networks, we seek to systematically characterize the multimodal network disruption patterns in aMCI.Methods45 aMCI patients and 42 healthy adults from the First Affiliated Hospital of Heilongjiang University of Chinese Medicine in Harbin, Heilongjiang Province, China, were enrolled. A case-control cross-sectional study was conducted. DTI and rs-fMRI data were collected for all participants. Global topological properties of structural and functional networks were constructed using PANDA and dpabi software and were calculated via graph-theoretical analysis in GRETNA software, followed by statistical comparisons between groups.ResultsIn patients with aMCI, the small-world (Cp, aCp, Lambda, aLambda) of the WM structural network were significantly higher than those in the HC group; Rich-club nodes showed redistribution, and the Rich-club coefficient was decreased; aEloc was significantly increased; the Assortativity index (r < 0) indicated disassortativity; the Hierarchy index (b > 0) exhibited a significant decrease in b within the sparsity range of 0.39∼0.4; the synchronization coefficient (s) was significantly reduced at sparsity levels ranging from 0.28 to 0.30. For the functional network, the small-world index aLp in the aMCI group was significantly lower than that in the HC group; Rich-club nodes showed redistribution, and the Rich-club coefficient was increased within a certain Degree range; aEg was significantly increased; the Assortativity index (r > 0) indicated assortativity; the Hierarchy index (b > 0) was observed within a specific sparsity range.ConclusionWe identified a “structure-function dissociation” in aMCI, where the structural network suffers from fragmentation and hub disruption, while the functional network compensates through rigid, hyper-localized reorganization with elevated local efficiency. This divergence reveals a core pathological mechanism of the disease.

The hybrid modular multilevel converter based on full-bridge and half-bridge submodule is a promising candidate for medium- and high-voltage applications. But different from conventional two-level converter, the power loss distribution inside submodules is uneven which will threaten the reliability. In this article, a reliability assessment model considering the device-level uneven power losses is proposed. Then influence of power loss distribution and redundancy schemes are investigated through a case study. The results illustrate that more unbalanced power loss distribution will result in reduced reliability. And improved modulation strategy can reduce the loss imbalance inside submodules. It is also found that passive redundancy scheme works better than active redundancy scheme in improving reliability as the failure rates of bypassed submodules are much lower than operating submodules. And passive scheme with 20% redundancy rate of full-bridge submodules is a good choice for redundancy design which both meets 20-year B10 lifetime requirement and saves cost.

Based on the principle of reasoning, this paper studies and analyzes the problems and causes of traditional teaching methods that tend to appear in reasoning course education. Taking folk art as an example, this paper puts forward the classification and deductive teaching methods, and provides valuable references to optimize methods in education and teaching.

目的 探讨颈椎邻椎病患者翻修手术方式及疗效.方法 对2007年至2014年我院收治的7例颈椎前路融合术后出现邻椎病而行翻修手术治疗的患者进行回顾分析.其中颈椎前路翻修手术患者3例,后路翻修手术患者4例.采用改良JOA评分(27分法)及VAS评分进行评价.结果 颈椎前路间盘切除融合术治疗患者3例,翻修术后内固定物位置良好,随访未见钢板螺钉松动,植骨块移位等;颈椎后路单开门椎管扩大成形术治疗患者4例,翻修术后无感染等并发症发生.与术前相比,颈椎前路和后路患者在术后随访结束时颈椎JOA评分均明显升高,VAS评分均明显减少.结论 颈椎邻椎病翻修术式的选择应遵循兼顾减压及尽可能保留颈椎活动度的原则,翻修术后可取得满意的疗效.

Objective To investigate the breastfeeding knowledge and behavior of the children's mother and network utilization status.Methods Children's mothers from two outpatient departments in Xicheng district were sampled and conducted questionnaire survey.Results Among the surveyed 215 children's mothers,56.3% of them did not know or could not correctly understand the concept of exclusive breastfeeding.Merely 27.9% of the subjects knew the exact duration of exclusive breastfeeding.Approximately 32.3% of the children's mothers' knowledge and behavior were not uniform in terms of the infant exclusive breastfeeding.Approximately 87.4% of the people paid attention to the knowledge about infant and young child feeding on the internet,and 69.9% of them tended to use the search engine.After acquired the solution on the internet,90.5% of them would choose to adopt.Conclusion The mothers' knowledge about exclusive breastfeeding was poor,calling for strengthened popularization of the feeding knowledge;and the internet was one of the main sources of infant feeding knowledge.

Objective To explore surgical technique for the refractory epilepsy (RE) induced by non-specific chronic encephalitis (NSCE) and curative effect of surgery on RE. Methods The clinical data of 6 patients with RE induced by NSCE, who underwent surgery under monitoring intraoperative electrocephalogram (EEG) from April, 2008 to August, 2010, were analyzed retrospectively, including clinical manifestations, EEG, imaging data, surgical method, curative effect and so on. Results EEG was mildly or moderately abnormal in all the patients. The encephalodialysis in 3 patients, the abnormal signal in 2 patients and no abnormal findings in 1 patient were shown by MRI. The resection of the epileptogenic foci in 2 patients, the resection of the epileptogenic foci plus amygdalo-hippocampectomy in 2 patients and resection of the epileptogenic foci plus multiple subpial transection in 2 patients were performed. The pathological examination showed that there was non-specific chronic encephalitis in all the patients, there was glial cell proliferation in 4 patients and there was focal cortical dysplasia in 3 patients. Of 6 patients, 2 had Engel grade Ⅰ therapeutic outcome, 1 grade Ⅱ and 2 grade Ⅲ , 1 grade Ⅳ . Conclusion The curative effect of surgery on the refractory induced by NSCE is not ideal, especially in the patients with refractory epileptic state.

Objective To observe the effect of Nogo-p4 on process growth of bipolar astrocyte differentiated from neural stem cells ( NSCs ). Methods Neurospheres were collected by suspension culture of cells obtained from spinal cords of 4 Wistar rats born in 24 hours. Immunofluorescence method was used to identify NSCs and detect expression of glial fibrillary acidic portein (GFAP) after NSCs differentiated. The NSCs were divided into groups A and B. FBS was added to group A and FBS + Nogo-p4 was added to group B to initiate glial differentiation. The process length of bipolar astrocytes was measured. Results Seven days after differentiation, bipolar astrocytes were observed in both groups. The length of the process on bipolar astrocyte in group B was shorter than that in group A significantly. Conclusion The process growth of bipolar astrocytes differentiated from NSCs derived from the spinal cord of rats is inhibited significantly by Nogo-p4.

Objective: To observe the effect of Nogo-p4 on radial differentiation of neural stem cells.Methods: Neural stem cells were cultured in vitro and divided into A and B group.Neural stem cells of group A differentiated at the presence of bovine serum.Nogo-p4(4μmol/L) was added to the medium of group B during the differentiation progress of neural stem cells.GFAP was used to mark the astrocyte by immunocytoflurecence method.Astrocyte subtype percentage after differention was observed.Results: Seven days after differentiation,four subtype of astrocyte were observed in group A: elongated,flat and polygonal,star like and bipolar type.In group B all the subtypes of astrocyte could be seen except the flat and polygonal subtype.Conclusion: Nogo-p4 specifically inhibited the flat and polygonal astrocyte subtype during the differentiation progress of neural stem cell.

Objective To observe the subtype of astrocyte differentiated from neural stem cells(NSCs) from spinal cord of rats.Methods Neurospheres were collected by suspension culture of cells obtained from rat spinal cord.Immunofluorescence method was used to identify NSCs and detect expression of GFAP after NSCs differentiated.The morphological character and ratio of each subtype was observed.Results Seven days after differentiation,four subtype of astrocyte were observed: elongated,flat and polygonal,star like and bipolar type.The ratio of each subtype was 2.39%±0.13%,32.33%±2.01%,59.88%±3.12% and 5.39%±0.27% respectively.Conclusion Astrocytes differentiated from neural stem cells from spinal cord of rats NgR has multiple subtypes.

Objective To observe the effect of Nogo-p4 on process growth of bipolar astrocyte differentiated from neural stem cells(NSCs) with and without the silenced NgR gene. Methods Neurospheres were collected by suspension culture of cells obtained from spinal cords of 4 wistar rats born in 24 hours.Immunofluorescence method was used to identify NSCs and detect the expression of GFAP after NSCs differentiated.The NSCs were divided into groups A,B,C and D.FBS was added to group A to initiate glial differentiation;FBS+Nogo-p4 was added to group B;in group C NSCs were transfected by siRNA to knockdown NgR before they differentiated;FBS+Nogo-p4 was added to group D after siRNA transfection and silencing of the NgR gene.The process length of bipolar astrocytes was measured by Image-Pro Plus 5.0 software. Results Seven days after differentiation,bipolar astrocytes were observed in each group.The length of the processes on bipolar astrocytes in group B was significantly shorter than that in other groups. Conclusion In combination with NgR,Nogo-p4 inhibits the process growth of bipolar astrocytes differentiated from NSCs derived from spinal cords of rats.

OBJECTIVE:To discuss the stability and practicality of temporomandibular joint replacement by establishing goats artificial temporomandibular joint replacement model. METHODS:Six healthy mature goats were selected, the male and female being half and weighing 35.3-37.0 kg. According to the parameters from X-ray films of goat's temporomandibular joint and the shape of the same kind goat's skull, the total temporomandibular joint prosthesis was prepared. The one side temporomandibular joints of six goats were replaced by prosthesis randomly as the experimental group (n = 6, fossa and condyle according to replacement location) and the other side by titanium plate as the control group (n = 6). At 4, 8, and 12 weeks, the histological observation, scanning electron microscope (SEM) observation were carried out for observing structural changes in the interface. The mechanical test and histochemistry test were used for observing the combination degree of interface and the alkaline phosphatase (ALP) activity. RESULTS:All animals were alive to the end of experiment with normal open mouth, good recovery of masticatory function, and normal eating. At 4, 8, and 12 weeks, implants were stable in 2 groups without loosening. The histological observation and SEM observation showed the amount of osteoblasts in interface increased over times. There were significant differences in the shearing force and the ALP activity between fossa in experimental group and control group at 4 weeks (P < 0.05), but there was no significant difference between other groups (P > 0.05). CONCLUSION:The total temporomandibular prosthesis has good stability in temporomandibular joint reconstruction of goat after replacement.

Objective:We investigate whether replication-competent adenovirus Ad5(E1+E3+) could enhance the expression of hCNTF secreted by celllines after replication-defective adenovirus Ad5-hCNTF infection in vitro.Methods:Plasmid encoding human CNTF was digested and subcloned into shuttle plasmid pshuttle to obtain a recombinant plasmid pshuttle-hCNTF.Using Ad-EASY system,the lineared and dephosphorylated shuttle vector and pAdEasy-1 plasmid which contain most of the human adenovirus serotype 5 genome were cotransfomed into BJ5183 bacteria,a recombination event then take place and result in the production of recombinant AdEasy plasmid,then the plasmid carrying adenovirus major genome and target gene was transfected into package cells AD-293 celllines to obtain a replication-defective adenovirus Ad5-hCNTF.Results:The recombinant adenovirus Ad5-hCNTF were analyzed by PCR and the expression of hCNTF in retinal pigment epithelium(RPE) cells after Ad infection were examined by Western blotting,and the ability of the cell to secrete CNTF was measured with a sandwich enzyme-linked immunosorbent assay(ELISA).the results indicated that Ad5-hCNTF contained the target gene,and were capable of infecting RPE celllines and produce the protein into the supernatant.Conclusion:We also have demonstrated that combined with different amount of replication-competent adenovirus Ad5(E1+E3+) in vitro,the expression of hCNTF secreted by celllines after Ad infection was enhanced significantly,and the enhancement of gene expression was observed in a concentration-dependent manner,and it offers basis for further study of gene therapy by Ad.

BACKGROUND AND OBJECTIVEAdenovirus vectors were widely used in gene therapy for tumors. We used adenovirus vector to transfer small interfering RNA (siRNA) against vascular epithelium growth factor A (VEGF-A) molecules to mouse lung adenoma LA795 cells and used low dose of chemotherapeutic drugs to further elevate the infection efficiency of adenovirus vector in and therapeutic effect of RNAi on tumor cells.METHODSLA795 cells were infected by Ad/EGFP and treated with different dosages of gemcitabin, epirubicin, cisplatin, or 5-fluorouracil (5-FU). Cells were observed under fluorescence microscope continuously using green fluorescent protein (GFP) as the reporter gene. The percentage of GFP-positive cells and fluorescent intensity were tested by flow cytometry to determine optimum concentrations of drugs. Ad/siVEGF-A containing VEGF-A siRNA was constructed. Real-time PCR and ELISA were applied to measure the expression level of VEGF-A after LA795 cells were infected by Ad/siVEGF-A and treated with 5-FU. The combination of Ad/siVEGF-A and 5-FU was also applied in treating subcutaneous tumor in mice.RESULTSLow dose of 5-FU elevated the Ad/EGFP infection in LA795 cells significantly, and also enhanced the effect of Ad/siVEGF-A in down-regulating VEGF-A mRNA and protein levels in tumor cells. When used in tumor in vivo, the combination strategy repressed tumor growth effectively.CONCLUSIONLow dose of 5-FU can enhance the capability of adenovirus infecting tumor cells and promote the efficiency of gene therapy by adenovirus.

Objective:On the treatment of regression knee joint disease by traditional Chinese medicine(Xikang),studying the treatment effect of Chinese medicine,to determine the clinical value of traditional Chinese medicine on regression knee joint disease.Methods:34 cases(40 knees)had regression knee joint disease,by intra-articular injection of knee with the Xikang solution,in 16 weeks,contrast observation was done before and after treatment.Results:The effects of treatment were compared in 34 cases,the physical signs,such as the joint pain,the greatest walk distance,joint function,daily active ability and joint swelling were superior to those before(P<0.05).Conclusion:This medicine had clearer effect,higher security and fewer side effect,and it was a kind of safe and effective medicine on the observation of the therapeutic effect of regression knee joint disease.

Objective To observe the kinesis expression of the expression of nestin ,nuclear factor-kB (NF-kB) and glial fibrillary acidic protein (GFAP) in the CA3 area of hippocampus of the experimental posttraumatic epilepsy (PTE) rats,and discuss the possible pathogenesis of PTE. Methods Twenty-six healthy adult male Sprague-Dawley rats were randomized into normal control group (A,n=6) with no treatment and epilepsy model group(B,n=20) with 1μmol FeCl2 injection into the motor cortex. Behavior was observed and EEG were recorded,brains were removed at different time points. The expression of nestin,NF-kB and GFAP in the CA3 area of hippocampus were detected immunohistochemically to observe the pathological changes. Results 15 rats of group B were survived after the injections. Compared with group A,the expression of nestin in the CA3 area of hippocampus of group B gradually increased at one day after injection and increased significantly 7 days later(P<0.05),and decreased after 14 days. The expression of NF-kB sharply increased one day after injection,decreased in 7 days later and significantly decreased after 14 days. The expression of GFAP increased obviously after 7 days and this high expression level was maintained till 14 days compared to group A(P<0.05). Conclusion The expression of nestin increased gradually after PTE,and the highest expression of NF-kB of neurons and glial cells appeared early. The proliferation and differentiation of neural precursor cells were induced after PTE. Moreover,the self-proliferation and hyperplasia of glial cells may play an important role in the epileptogenesis of PTE.

The transcription factor E2F1 may form a complex with RB and which may play a major role in promoting growth control. Small interfering RNAs are short, double-stranded RNA molecules that can target mRNAs with complementary sequence for degradation via a cellular process to induce gene silence through degradation of homologous transcripts. The experiments were designed to inhibit proliferation of human retinoblastoma cells by knockdown human transcription factor E2F1 expression level using RNA interference technology. Three different hairpin shRNA inserts were designed targeting human transcription factor E2F1. The three inserts were synthesized and subcloned into a shRNA espression vector pSilencer H1puro which contains the HI RNA polymerase III promoter. The three plasmids encoding E2F1 short hairpin were transfected into human retinoblastoma cells line HXO-Rb-44 and human liver cancer cell line SMMC-7721 in vitro. Real-time RT-PCR was used to measure E2F1 mRNA level. For function analysis, propidium iodide (PI)-stained DNA content method was used to identify cell cycle by flow cytometer. The selected shE2F1 were cloned into an adenovirus system and packaged to adenovirus. Retinoblastoma HXO-Rb-44-gfp cell line which has been marked with green fluorescent protein (GFP) were infected by Ad-shE2F1 for 48h, and Ad-shNegative was used as control. The infected cells were injected into anterior chamber of nude mice eyes. The eyes were investigated and the consecutive and dynamic photos of the condition of RB tumor's growth were taken by the special fluorescent microscope. One in three RNAi expression plasmids delivery resulted in 83.5% suppression of E2F1 mRNA transcription level. The same shRNA showed 89% inhibition in entering S phase. In nude mice eye anterior chamber, Ad-shE2F1 infected tumor cells grew more slowly than the negative control. The results showed that efficacy of Ad-shE2F1 were helpful in the process of limiting the growth of retinoblastoma. RNAi gene therapy directed against the human transcription factor E2F1 is a new therapeutic approach to suppress retinoblastoma cells proliferation.

[Objective]To discuss the clinical results of reconstruction of block bone deficit knee(lower femur and upper tibia) by transplantation of patella and present surgery protocols.[Methods]There were 7 patients who have giant cell tumor of bone in the lower femur or upper tibia at the knee in this report.The case history was from 4 to 12 months,average 5.8 months.Four were male and three were female.The age ranged from 18 to 40 years,average 31.8 years.Four had lesions at the lower femur and the other three had lesions at the upper tibia.According to Ennecking grade,five cases were in grade 2 and two cases were in grade 3.When the cartilage of the knee was damaged by bone tumors,the author supplemented it by autogenous patella to fill the defect.[Results]All cases of patella grafts were presented,with follow-up from 3 to 10 years(average 4.9 years),three cases were followed up more than 5 years.During the follow up,there was no case with recurrence、metastasis or necrosis and infection of graft.All the grafts had good union.According to the function evaluation standard of the international protecting extremities institute,3 excellent,4 good.The main complications were less range of movement of the knee(movement range 40～110 degree,average 88 degree).Weakness of the extensor apparatus was not obvious.One case had slight collapse of tibia plateau and another case sufferd from joint pain 6 years after operation.[Conclusion]When the cartilage of the lower femur and the upper tibia is damaged by bone tumors,the bone tumors and its affected condyle cowld be incised,then apply patella grafts to repair and reconstruct the defect of the knee.

Objective To observe the effect of RNAi on the NgR gene expression of neural stem cells derived from the rat spinal cord in vitro.Methods Three siRNAs were disigned and synthesized according to the mRNA sequence of rat NgR gene,and transfect to neural stem cells in culture.The NgR gene expression was detectecl by immunofluoresence and Western blot.Results The NgR gene was knocked down by the three siRNAs in different degrees.The knock-down efficiency decreased with time after transfection.The most effective siRNA kept high knock-down efficiency(85.22±3.1%) even on the day 5 after transfection.Conclusion The NgR gene in rat neural stem cells can be knocked down effectively by RNAi.

神经干细胞研究一直是神经科学领域的热点.目前国内外学者已经建立了神经干细胞的分离和培养技术[1].本研究分析大鼠出生后不同时间大鼠脊髓神经干细胞的培养特点,为应用神经干细胞积累经验.