Enhancing soil health and strawberry disease resistance: the impact of calcium cyanamide treatment on soil microbiota and physicochemical properties.

Introduction: Continuous strawberry cropping often causes soil-borne diseases, with 20 calcium cyanamide being an effective soil fumigant, pig manure can often be used as soil organic fertilizer. Its impact on soil microorganisms structure, however, remains unclear. Methods: This study investigated the effectiveness of calcium cyanamide and pig manure in treating strawberry soil, specifically against strawberry anthracnose. We examined the physical and chemical properties of the soil and the rhizosphere microbiome and performed a network analysis. Results: Results showed that calcium cyanamide treatment significantly reduces the mortality rate of strawberry in seedling stage by reducing pathogen abundance, while increasing actinomycetes and Alphaproteobacteria during the harvest period. This treatment also enhanced bacterial network connectivity, measured by the average connectivity of each Operational Taxonomic Unit (OTU), surpassing other treatments. Moreover, calcium cyanamide notably raised the levels of organic matter, available potassium, and phosphorus in the soil-key factors for strawberry disease resistance and yield. Discussion: Overall, applying calcium cyanamide to soil used for continuous strawberry cultivation can effectively decrease anthracnose incidence. It may be by changing soil physical and chemical properties and enhancing bacterial network stability, thereby reducing the copy of anthracnose. This study highlights the dual benefit of calcium cyanamide in both disease control and soil nutrient enhancement, suggesting its potential as a valuable tool in sustainable strawberry farming.

S100A9 Upregulation Contributes to Learning and Memory Impairments by Promoting Microglia M1 Polarization in Sepsis Survivor Mice.

Sepsis-associated encephalopathy (SAE) is a clinical syndrome of brain dysfunction secondary to sepsis, which is characterized by long-term neurocognitive deficits such as memory, attention, and executive dysfunction. However, the mechanisms underlying SAE remain unclear. By using transcriptome sequencing approach, we showed that hippocampal S100A9 was significantly increased in sepsis induced by cecal ligation and puncture (CLP) or lipopolysaccharide (LPS) challenge. Thus, we used S100A9 inhibitor Paquinimod to study the role of S100A9 in cognitive impairments in CLP-induced and LPS-induced mice models of SAE. Sepsis survivor mice underwent behavioral tests or the hippocampal tissues subjected to Western blotting, real-time quantitative PCR, and immunohistochemistry. Our results showed that CLP-induced and LPS-induced memory impairments were accompanied with increased expressions of hippocampal microglia Iba1 and CD86 (M1 markers), but reduced expression of Arg1 (M2 marker). Notably, S100A9 inhibition significantly improved the survival rate and learning and memory impairments in sepsis survivors, with a shift from M1 to M2 phenotype. Taken together, our study suggests that S100A9 upregulation might contribute to learning and memory impairments by promoting microglia M1 polarization in sepsis survivors, whereas S100A9 inhibition might provide a potential therapeutic target for SAE.

Interpretation of subjective cognitive decline characteristics published in Lancet Neurology / 中华神经科杂志

Alzheimer′s disease (AD) is an incurable disease in the field of major chronic diseases. Subjective cognitive decline (SCD) is a clinical risk factor for AD. The standardized screening and intervention in individuals with SCD are of great importance in early prevention and treatment of AD. According to the clinical criteria proposed by The characterisation of subjective cognitive decline, which was published online in Lancet Neurology, the article summarized the definition of SCD, the latest perspective of clinical standards in SCD, and the results of AD preclinical SCD research. The purpose of this work was to provide concrete guidance and recommendations for making clinical decisions in diagnosis and scientific research on SCD.

The effects of ApoE epsilon4 alleles on cognitive function and resting-state functional MRI in patients with amnestic mild cognitive impairment: a prospective cohort study / 中华放射学杂志

Objective:To explore the effects of ApoE epsilon4 (ApoE-ε4) alleles on cognitive function and resting-state functional MRI (rs-fMRI) in patients with amnestic mild cognitive impairment(aMCI) based on a prospective cohort study.Methods:An average of 20 months of prospective observations were conducted on 16 ApoE-ε4-carriers and 24 non-carriers of aMCI. Neuropsychological assessments and rs-fMRI data were collected at both baseline and follow-up. All participants were assessed by a battery of neuropsychological tests and underwent rs-fMRI. Two core regions of the default mode network (DMN), the left posterior cingulate cortex (PCC) and the medial prefrontal cortex (mPFC), were selected as seeds to calculate the functional connectivity. Two-way repeated measures analysis of variance was used to assess the effects of ApoE genotype(ε4-carriers, nonε4-carriers), interval and the interaction between these two factors for functional connectivity extracted from changed region found by t-test.Conversion rates of dementia were compared between ApoE-ε4-carriers and nonε4-carriers at follow-up using Chi-square test. For the comparison of functional connectivity and clinical data between ApoE-ε4-carriers and nonsε4-carriers in baseline and follow-up, the normal distribution test was carried out first. If the normal distribution was fitted, the two-sample t test was used, otherwise, the Mann-Whitney rank sum test was used. Finally, the general linear model was used to assess the relationships between alterations in functional connectivity and in neuropsychological assessments as well as the interaction effect. Results:(1)Significant decline in memory domains were found in ApoE-ε4-carriers as compared to non-carriers at both baseline and follow-up. The ApoE-ε4-carriers (14/16) presented a higher conversation rate than non-carriers(13/24, χ 2=4.862, P=0.027) at follow-up. (2)Functional imaging analysis revealed that ApoE-ε4-carriers exhibited significantly higher functional connectivity between the left PCC and the left angular (ApoE-ε4-carriers: 0.23±0.11, non-carriers: -0.03±0.13, t=4.800, cluster size: 1 944 mm 3, P=0.004), and between the left mPFC and the left angular (ApoE-ε4-carriers: 0.33±0.21, non-carriers: 0.08±0.18, t=5.040, cluster size:1 836 mm 3, P=0.006) as compared to non-carriers at follow-up. We detected significant effect for the interaction interval by ApoE-ε4 on functional connectivity between the left angular and the left PCC ( F=10.833, P=0.002)as well as the left mPFC ( F=7.280, P=0.010). (3)The alteration of functional connectivity value between the left mPFC and the left angular in ApoE-ε4-carriers was positively correlated with the changes ofimmediate memory ( r=0.692, P=0.018). The correlation was not statistically significant in ApoE-ε4-noncarriers ( r=-0.198, P=0.417) and the integration effect was significant ( F=8.632, P=0.006). Conclusions:The ApoE-ε4 actually accelerates the deterioration of cognitive function in aMCI patients and carriers presented relatively reserved functional connectivity between the left angular and other core regions within DMN, which indicated the disruption of functional connectivity may be one of the underline mechanisms of ApoE-ε4 during AD process.

The effects of ApoE epsilon4 alleles on cognitive function and resting-state functional MRI in patients with amnestic mild cognitive impairment: a prospective cohort study / 中华放射学杂志

Objective@#To explore the effects of ApoE epsilon4 (ApoE-ε4) alleles on cognitive function and resting-state functional MRI (rs-fMRI) in patients with amnestic mild cognitive impairment(aMCI) based on a prospective cohort study.@*Methods@#An average of 20 months of prospective observations were conducted on 16 ApoE-ε4-carriers and 24 non-carriers of aMCI. Neuropsychological assessments and rs-fMRI data were collected at both baseline and follow-up. All participants were assessed by a battery of neuropsychological tests and underwent rs-fMRI. Two core regions of the default mode network (DMN), the left posterior cingulate cortex (PCC) and the medial prefrontal cortex (mPFC), were selected as seeds to calculate the functional connectivity. Two-way repeated measures analysis of variance was used to assess the effects of ApoE genotype(ε4-carriers, nonε4-carriers), interval and the interaction between these two factors for functional connectivity extracted from changed region found by t-test.Conversion rates of dementia were compared between ApoE-ε4-carriers and nonε4-carriers at follow-up using Chi-square test. For the comparison of functional connectivity and clinical data between ApoE-ε4-carriers and nonsε4-carriers in baseline and follow-up, the normal distribution test was carried out first. If the normal distribution was fitted, the two-sample t test was used, otherwise, the Mann-Whitney rank sum test was used. Finally, the general linear model was used to assess the relationships between alterations in functional connectivity and in neuropsychological assessments as well as the interaction effect.@*Results@#(1)Significant decline in memory domains were found in ApoE-ε4-carriers as compared to non-carriers at both baseline and follow-up. The ApoE-ε4-carriers (14/16) presented a higher conversation rate than non-carriers(13/24, χ2=4.862, P=0.027) at follow-up. (2)Functional imaging analysis revealed that ApoE-ε4-carriers exhibited significantly higher functional connectivity between the left PCC and the left angular (ApoE-ε4-carriers: 0.23±0.11, non-carriers: -0.03±0.13, t=4.800, cluster size: 1 944 mm3, P=0.004), and between the left mPFC and the left angular (ApoE-ε4-carriers: 0.33±0.21, non-carriers: 0.08±0.18, t=5.040, cluster size:1 836 mm3, P=0.006) as compared to non-carriers at follow-up. We detected significant effect for the interaction interval by ApoE-ε4 on functional connectivity between the left angular and the left PCC (F=10.833, P=0.002)as well as the left mPFC (F=7.280, P=0.010). (3)The alteration of functional connectivity value between the left mPFC and the left angular in ApoE-ε4-carriers was positively correlated with the changes ofimmediate memory (r=0.692, P=0.018). The correlation was not statistically significant in ApoE-ε4-noncarriers (r=-0.198, P=0.417) and the integration effect was significant (F=8.632, P=0.006).@*Conclusions@#The ApoE-ε4 actually accelerates the deterioration of cognitive function in aMCI patients and carriers presented relatively reserved functional connectivity between the left angular and other core regions within DMN, which indicated the disruption of functional connectivity may be one of the underline mechanisms of ApoE-ε4 during AD process.

Research on characteristics of resting-state functional connectivity strengths in patients with amnestic mild cognitive impairment / 中华放射学杂志

Objective To explore a new index for reflecting the topological information of brain functional networks in patients at high risk of Alzheimer disease using characteristics of resting-state functional connectivity strengths(FCS) in patients with amnestic mild cognitive impairment(aMCI). Methods Thirty-one aMCI patients and 42 age, gender and years of education matched normal controls were enrolled between September 2009 and April 2011 in this study. The resting-state functional MRI (rs-fMRI) data of all participants were acquired and preprocessed. Then the whole-brain functional connectivities were constructed for exploring the distribution characteristics of hub regions which had higher FCS values. Using two-sample t test to compare group differences in age, years of education and each neuropsychological assessment. In addition, using Chi-squared test to compare group differences in gender. Group differences in FCS values were analyzed by general linear model. Finally, correlation analyses were used to evaluate the relationships between the FCS values of the brain regions with group differences and behavioral scores in aMCI patients. Results The hub regions of the functional networks in the aMCI patients were mainly located in the association cortices such as the precuneuses, posterior cingulate cortices, medial prefrontal cortices, angular gyri, superior occipital gyri, fusiform gyri and lingual gyri. The distribution models in the aMCI patients were consistent with those in the normal controls. However, the FCS values of these brain regions were significantly lower in the aMCI patients than those in the normal controls. In comparison to the normal controls, the aMCI patients had significantly decreased FCS values in the bilateral fusiform gyri, lingual gyri, superior occipital gyri, left middle occipital gyrus and postcentral gyrus (the cluster was 389, 230, 187 and 107 voxels, respectively;P<0.05, respectively), and they had decreased trends of FCS values in the bilateral posterior cingulate cortices and right insulas. The correlation analysis with uncorrected conditions showed that the FCS values of the left postcentral gyri were correlatid with the clock drawing test (CDT) scores (r=0.436, P=0.026). Conclusions aMCI mainly attacks the hub regions of brain functional networks. The changes of functional connectivities in aMCI may reflect the early pathophysiologic alterations of AD.

Progress of biomarkers for subjective cognitive decline / 医学研究生学报

Along with the development of biomarkers, the diagnostic criterion for early AD is continuously progressing until the preclinical stage of AD, on the base of which, the conception of subjective cognitive decline was raised.In order to highlight new ideas of the early diagnosis for AD in its preclinical stage, the current paper will talk about SCD in connection with neuroimaging tech-niques and examination of cerebrospinal fluid.

Simvastatin induces neuroprotection in 6-OHDA-lesioned PC12 via the PI3K/AKT/caspase 3 pathway and anti-inflammatory responses.

BACKGROUND: In addition to their original applications for lowering cholesterol, statins display multiple neuroprotective effects. Inflammatory reactions and the PI3K/AKT/caspase 3 pathway are strongly implicated in dopaminergic neuronal death in Parkinson's disease (PD). This study aims to investigate how simvastatin affects 6-hydroxydopamine-lesioned PC12 via regulating PI3K/AKT/caspase 3 and modulating inflammatory mediators. METHODS: 6-hydroxydopamine-treated PC12 cells were used to investigate the neuroprotection of simvastatin, its association with the PI3K/AKT/caspase 3 pathway, and antiinflammatory responses. Dopamine transporters (DAT) and tyrosine hydroxylase (TH) were examined in 6-hydroxydopamine-treated PC12 after simvastatin treatment. RESULTS: Simvastatin-mediated neuroprotection was associated with a robust reduction in the upregulation induced by 6-OHDA of inflammatory mediators including IL-6, COX2, and TNF-α. The downregulated DAT and TH levels in 6-OHDA-lesioned PC12 were restored after simvastatin treatment. Simvastatin reversed 6-OHDA-induced downregulation of PI3K/Akt phosphorylation and attenuated 6-OHDA-induced upregulation of caspase 3 in PC12. Furthermore, the PI3K inhibitor LY294002 pronouncedly abolished the simvastatin-mediated attenuation in caspase 3. CONCLUSIONS: Our results demonstrate that simvastatin provides robust neuroprotection against dopaminergic neurodegeneration, partially via antiinflammatory mechanisms and the PI3K/Akt/caspase 3 pathway. These findings contribute to a better understanding of the critical roles of simvastatin in treating PD and might elucidate the molecular mechanisms of simvastatin effects in PD.

Transforming growth factor-ß genetic polymorphisms on development of liver cirrhosis in a meta-analysis.

Transforming growth factor-ß (TGF-ß) protein has been supposed to be a risk factor for liver cirrhosis; however, the associations between its genes (TGF-ß -509C>T and +869T>C) and liver cirrhosis remained unclear. This study was to quantitatively analyze the correlations by using a meta-analysis. Pubmed, Embase, Wanfang databases were retrieved up to November 1st, 2011. Odds ratio (OR) and 95 % confidence interval (95 %CI) were used to demonstrate the strength of association, and P < 0.05 of Z test indicated statistical significance. Combined analyses were performed by using fixed or random-effect model, depending on between-study heterogeneity. Seven studies were for TGF-ß -509C>T polymorphism, and eight studies were for +869T>C polymorphism. Combined results indicated that neither TGF-ß -509C>T nor +869T>C polymorphisms were associated with risk of liver cirrhosis [OR (95 % CI): 0.79 (0.60-1.04) for CT vs. TT of -509C>T and 0.87 (0.68-1.12) for CT vs. CC of +869T>C], with no between-study heterogeneity. In addition, subgroups analyses still inferred that two polymorphisms were not associated with risk of liver cirrhosis for HBV-infected patients, Asians and for Population-based studies. This meta-analysis indicated that neither TGF-ß -509C>T nor +869T>C polymorphisms were associated with risk of liver cirrhosis, regardless of HBV infection or not.

[Analysis of the correlation between parapapillary atrophy zones and visual field loss].

OBJECTIVE: To analyze the association between parapapillary atrophy zones and mean sensibility (MS) and mean defect (MD) from visual field examination. METHODS: Selection was performed among 201 cases (402 eyes) who underwent Heidelberg retinal tomography (HRT) and visual field examination (OCTOPUS) during the 2-week duration at Department of Ophthalmology, Peking University People's Hospital between April 2001 to October 2002. Of all the cases, 103 (161 eyes) that had suffered from MD>2 dB and whose refractive error and fundus appearance did not meet exclusion criteria were enrolled. RESULTS: Maximum diameters of ß zone and α zone were not significantly associated with refractive errors, disc area, optic cup area and rim area (all P>0.05). Maximum diameters of both ß zone (r=-0.188, P=0.017)and α zone(r=0.209, P=0.008) were significantly associated with MS. Similarly, maximum diameters of both ß zone (r=0.159, P=0.044)and α zone(r=-0.191, P=0.015) were significantly associated with MD. However, the maximum diameter ofß zone was the risk factor to defect of visual field, but the maximum diameter of α zone was the protective factor to defect of visual field. CONCLUSION: Among the patients with remarkable defect of visual field, the maximum diameters of parapapillary atrophy zones are associated with damage of visual field.

Changes of interleukin-2 and interferon-γ in serum and synovial fluid of patients with joint injure with Kashin-Beck disease / 中国地方病学杂志

Objective To investigate the changes of interleukin(IL)-2 and interferon(IFN)-γin serum and synovial fluid of patients with joint injure with Kashin-Beck disease(KBD), and study the roles of IL-2 and IFN-γ in KBD joint injure. Methods In accordance with the "Diagnostic Criteria of Kashin-Beck disease"(GB16003-1995),48 cases of KBD patients and 26 healthy people(control group) from KBD endemic area in Long county Shaanxi province were enrolled in the study. KBD patient were 24 males and 24 females, respectively, aged 40 to 65 years (mean age 51 years). Forty-eight serum specimens and 28 synovial fluid specimens of patients(14 males and 14 females,respectively) were collected. Healthy control group were 13 males and 13 females, respectively. Twenty-six serum specimens of healthy controls were collected. Serum and synovial fluid IL-2 and IFN-γ levels were determined by enzyme-linked immunosorbent assay(ELISA). Results In healthy controls and KBD patients, the midian of serum IL-2 were 46.8 ng/L and 55.7 ng/L, respectively, and IFN-γ were 52.3 ng/L and 48.8 ng/L, respectively. The difference was not statistically significant between healthy controls and KBD patients(t = 0.62, 0.70, all P ＞ 0.05).In synovial fluid of KBD patient, the midian of IL-2 and IFN-γwere 48.3 ng/L and 44.1 ng/L, respectively. The difference was not statistically significant between serum and synovial fluid in KBD patients(t = 0.69, 1.72, all P ＞0.05). Conclusion Serum and synovial fluid IL-2 and IFN-γare not significantly increased in KBD patients with articular damage, indicating that IL-2 and IFN-γare not involved in KBD joint injury.

[Bone marrow stromal cells transfected with ciliary neurotrophic factor gene ameliorates the symptoms and inflammation in C57BL/6 mice with experimental allergic encephalomyelitis].

OBJECTIVE: To investigate the anti-inflammatory effect of bone marrow stromal cells (MSCs) transfected with recombinant adenovirus-mediated ciliary neurotrophic factor (CNTF) gene in C57BL/6 mice with experimental allergic encephalomyelitis (EAE). METHODS: An adenovirus vector containing CNTF gene Ad-CNTF-IRES-GFP was constructed and transfected in the MSCs (MSC-CNTF). After examination of CNTF expression, the transfected cells were transplanted in C57BL/6 mice with MOG 35-55-induced EAE, which were monitored for the changes in the symptoms scores. The levels of tumor necrosis factor-alpha (TNF-alpha), inteferon-gamma (IFN-gamma), interleukin-12P35 (IL-12P35), and IL-10 in the peripheral blood of the mice were detected, and the number of MSC-CNTF cells in the spleen and spinal cord was counted. CD3+ T cell infiltration and TNF-alpha and IFN-gamma expressions in the lesions were also observed after the cell transplantation. RESULTS: CNTF gene transfection resulted in significantly increased CNTF expression in the MSCs. The mice receiving MSC-CNTF transplantation exhibited significantly improved symptoms with shortened disease course and lessened disease severity. The cell transplantation also resulted in significantly decreased peripheral blood TNF-alpha levels, ameliorated CD3+T cell infiltrations and lowered TNF-alpha expression in the lesions, while the levels of IFN-gamma underwent no significant changes. CONCLUSION: Transplantation of CNTF gene-transfected MSCs results in decreased peripheral blood TNF-alpha and IFN-gamma levels and reduced inflammatory cells, CD3-positive cells and TNF-alpha expression in the lesion of EAE, therefore providing better effect than MSCs in relieving the symptoms of EAE in mice.

Investigation of drinking water fluoride and fluorosis in Shaanxi Province from 2005 to 2007 / 中国地方病学杂志

Objectives To investigate fluoride in drinking waters and fluorosis status and evaluate the effectiveness of fluoride-reducing projects in Shaanxi Province from 2005 to 2007. Methods In the Shaanxi province-wide, the 10 endemic areas of fluorosis were chosen according to historical data as focusing areas for investigation. The village was considered as investigation spot, 5 water samples were collected from each village for investigating of fluoride content. Four water samples were collected from each fluoride-reducing project for evaluating its effectiveness. Fluoride concentrations in drinking water were measured by fluoride-selective electrode method or speetrophotometry. When fluoride content in drinking water was greater than 1.00 mg/L, the epidemical study wasd conducted to investigate fluorosis patients, focusing on investigating of dental fluorosis prevalence in 8 to 12-year-old children and skeletal fluorosis prevalence in adults. Dental fluorosis of children was diagnosed by using Dean's method, and adult skeletal fluorosis was diagnosed according to National Standard for Clinical Diagnosis of Endemic Skeletal Fiuorosis(GB 16396-1996). Results The fluoride content in drinking water from 6390 villages was measured. The fluoride content of drinking water of 2619 villages ranged from 1.0 to<2.0 mg/L, where 1 654 998 people exposed. Additionally, the fluoride content of water of 845 villages ranged from 2.0 to<4.0 mg/L, where 355 623 people exposed. Moreover, the fluoride contents of water of 272 villages exceeded 4.0 mg/L, where 111 466 people exposed. The median of fluoride content in drinking water was 1.15% in the whole province, and fluoride content in drinking water exceeded 1.00 mg/L in Weinan, Xianyang and Yulin where were concentrated distribution areas of high fluoride water. Among 3115 fluoride-reducing projects, the fluoride content of drinking water of 1269 projects ranged from 1.0 to<2.0 mg/L, where 1 415 877 people exposed. Additionally, the fluoride content of drinking water of 120 projects ranged from 2.0 to<4.0 mg/L, where 43 888 people exposed. Moreover, the fluoride content of drinking water of 14 projects exceeded 4.0 mg/L, where 5960 people exposed. The detectable rate of dental fluorosis of 8 to 12 year-old children and skeletal fluorosis of adults reached 37.4%(16 489/44 081) and 5.1%(15 877/310 993), respectively. Conclusions The widely distribution of high-fluoride in drinking water still contributes to the prevalence of fluorosis in Shaanxi Province. The quality of fluoride-reducing projects should be further improved.

Bone marrow stromal cells transfected with ciliary neurotrophic factor gene ameliorates the symptoms and inflammation in C57BL/6 mice with experimental allergic encephalomyelitis / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the anti-inflammatory effect of bone marrow stromal cells (MSCs) transfected with recombinant adenovirus-mediated ciliary neurotrophic factor (CNTF) gene in C57BL/6 mice with experimental allergic encephalomyelitis (EAE).</p><p><b>METHODS</b>An adenovirus vector containing CNTF gene Ad-CNTF-IRES-GFP was constructed and transfected in the MSCs (MSC-CNTF). After examination of CNTF expression, the transfected cells were transplanted in C57BL/6 mice with MOG 35-55-induced EAE, which were monitored for the changes in the symptoms scores. The levels of tumor necrosis factor-alpha (TNF-alpha), inteferon-gamma (IFN-gamma), interleukin-12P35 (IL-12P35), and IL-10 in the peripheral blood of the mice were detected, and the number of MSC-CNTF cells in the spleen and spinal cord was counted. CD3+ T cell infiltration and TNF-alpha and IFN-gamma expressions in the lesions were also observed after the cell transplantation.</p><p><b>RESULTS</b>CNTF gene transfection resulted in significantly increased CNTF expression in the MSCs. The mice receiving MSC-CNTF transplantation exhibited significantly improved symptoms with shortened disease course and lessened disease severity. The cell transplantation also resulted in significantly decreased peripheral blood TNF-alpha levels, ameliorated CD3+T cell infiltrations and lowered TNF-alpha expression in the lesions, while the levels of IFN-gamma underwent no significant changes.</p><p><b>CONCLUSION</b>Transplantation of CNTF gene-transfected MSCs results in decreased peripheral blood TNF-alpha and IFN-gamma levels and reduced inflammatory cells, CD3-positive cells and TNF-alpha expression in the lesion of EAE, therefore providing better effect than MSCs in relieving the symptoms of EAE in mice.</p>

Adenoviral delivery of soluble VEGF receptor 1 (sFlt-1) inhibits experimental autoimmune encephalomyelitis in dark Agouti (DA) rats.

Previous studies have shown that vascular endothelial growth factor (VEGF) expression is up-regulated in both multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE), a model for MS, and may exacerbate the disease. However, it remains unknown whether anti-VEGF modalities could serve as a potential treatment for such central nervous system (CNS) autoimmune diseases. We constructed a recombinant adenoviral vector carrying FLAG-tagged sFlt-1(1-3) (the first three extracellular domains of Flt-1, the hVEGF receptor-1). Intramuscular transfection of the recombinant adenoviral vector suppressed VEGF-induced inflammatory cell infiltration in matrigel plugs. When given intracerebrally to EAE rats, recombinant sFlt-1(1-3) adenoviral vector significantly reduced disease severity compared to untreated rats. sFlt-1(1-3) gene transfer blocked VEGF and greatly reduced the number of cells that express VEGF and ED1-positive cells in CNS in EAE rats. This study demonstrates that sFlt-1(1-3) gene transfer into the brain ameliorates the severity of EAE by inhibiting monocyte recruitment in the CNS of dark Agouti rats.

Translocation and neuroprotective properties of transactivator-of-transcription protein-transduction domain-neuroglobin fusion protein in primary cultured cortical neurons.

Ngb (neuroglobin) is a newly discovered hexaco-ordinate globin that is expressed in vertebrate brain and peripheral nervous systems. Expression of Ngb increases in response to oxygen deprivation and protects neurons from hypoxia in vitro and in vivo. However, the lack of its transduction ability into cells resulted in limited neuroprotection. To educe its neuroprotection under hypoxia, a cell-permeable Ngb fusion protein was generated. A rat brain Ngb gene was cloned and fused with a gene fragment encoding the nine-amino-acid TAT PTD (transactivator-of-transcription protein-transduction domain; RKKRRQRRR) of HIV-1 in a prokaryotic expression vector to generate a genetic in-frame N-terminal hexahistidine-tagged) TAT PTD-Ngb fusion protein. It was expressed in soluble form in Escherichia coli BL21(DE3)plysS and purified with Ni(2+)-affinity chromatography. The results showed that the purified fusion protein TAT PTD-Ngb can enter into the primary cultured cortical neurons in a dose-dependent manner when added exogenously to the culture media and can be detected in cells within 48 h. The cell viability under hypoxia was increased and apoptosis induced by hypoxia was decreased after TAT PTD-Ngb was transduced into cortical neurons. The results provide a clue for the research of Ngb and suggest that transduction of TAT PTD-Ngb may be one of the ways for the therapy of CNS (central nervous system) diseases, especially cerebrovascular diseases and neurodegenerative diseases.

Identification and characterization of the CD226 gene promoter.

CD226 is one of the main activating receptors on natural killer cells, and it can induce cytotoxicity to target cells through interaction with its ligands CD155 or CD112. CD226 is also involved in T cell differentiation, activation, and cytotoxicity. The expression of CD226 on natural killer cells and T cells can be regulated by cytokines and chemical stimuli; however, the mechanism of the regulation of the CD226 gene is still unknown. In this study, we have identified two promoters in the human CD226 gene named P1 and P2, which are located at -810 to -287 bp and +33 to +213 bp, respectively, and a negative regulation element between P1 and P2. Both P1 and P2 can be regulated by phorbol ester (12-O-tetradecanoylphorbol-13-acetate) and calcium ionophore (A23187). Bioinformatics analysis shows that, within this CD226 gene region, there are putative binding sites for transcription factors AP-1, Sp1, PEA3, and Ets-1. We have found that transcription factor activating protein-1 (AP-1) can up-regulate CD226 promoters P1 and P2 in human hepatocarcinoma cells, a hepatocarcinoma cell line with low expression of endogenous AP-1 and Ets-1. Interestingly, the transcription factor Ets-1 promotes AP-1-induced P2 activity but inhibits AP-1-induced P1 activity for which a 10-bp AP-1/Ets-1 composite site (CCTTCCTTCC) in P1 may be responsible.

[Detection of proinflammatory cytokine levels in sera from patients with Kaschin-Beck disease].

AIM: To explore the role of proinflammatory cytokines (TNF, IL-1beta and IL-6) in the pathogenesis of Kaschin-Beck disease (KBD). METHODS: The levels of serum TNF, IL-1beta and IL-6 from 62 KBD patients and 60 healthy persons were detected by double antibody sandwich ELISA. RESULTS: The levels of serum IL-1beta and IL-6 in KBD patients were (238.4+/-698.5) ng/L and (164.4+/-661.4) ng/L, respectively, but they were higher than those in healthy persons, being (74.5+/-130.0) ng/L and (52.2+/-154.6) ng/L, respectively. There were no significant differences between them (P>0.05). However, the level of serum TNF in KBD patients [(109.2+/-145.3) ng/L] was significantly higher than that in healthy persons [(40.9+/-89.7) ng/L] (P<0.01). Moreover, there was no correlation between serum TNF and IL-1beta levels (r=0.0387, P>0.05) and TNF and IL-6 in KBD patients (r=0.2135, P>0.05), but there was positive correlation between serum IL-1beta and IL-6 levels (r=0.346, P<0.01). CONCLUSION: The elevated proinflammatory cytokine levels in sera may relate to the pathogenesis of Kaschin-Beck disease.

[Preparation and characterization of monoclonal antibodies against human RANTES molecule].

AIM: To prepare monoclonal antibodies against human RANTES molecule and identify the expression of RANTES in rat small intestine after small bowel transplantation. METHODS: Murine mAbs were prepared by B lymphocyte hybridoma technique. The expression of RANTES in rat small intestine after small bowel transplantation was detected by immunohistochemistry. RESULTS: Four hybridoma cell lines secreting monoclonal antibodies to human RANTES, FMU-RANTES 1, FMU-RANTES 2, FMU-RANTES 3 and FMU-RANTES 4, were established. The titers of a scetic mAbs reached to 1 x 10(-6) and the Ig subclass of FMU-RANTES 1, FMU-RANTES 3 and FMU-RANTES 4 was IgG1(kappa) and that of FMU-RANTES 2 was IgG2b(kappa). Among these mAbs, FMU-RANTES 1, FMU-RANTES 2 and FMU-RANTES 3 could bind human RANTES protein in Western bolt. FMU-RANTES 1, FMU-RANTES 2 and FMU-RANTES 4 could be used in immunohistochemistry staining. Rat RANTES molecule could be detected in the cyto plasm of epithelial cells in rat small intestine after small bowel transplantation. CONCLUSION: Four mAbs against RANTES molecule were prepared, which can provide a useful tool in research on the structure and function of RANTES molecule. High expression of RANTES may be involved in the rejection of allogeneic graft.

[The inhibition of TNF-induced NF-kappaB nuclear translocation in ECV304 cells by mAbs against human TNF].

AIM: To identify the inhibition of TNF-induced NF-kappaB nuclear translocation by three anti-human TNF mAbs, D2, E6 and F6. METHODS: TNF solutions were pretreated with mAbs D2, E6 and F6 as well as control mAb at 37 degrees Celsius for 1 h, respectively, and then they were added to ECV304 cell cultures. After 1 hour, the cells were harvested and nuclear proteins were extracted. The NF-kappaB activity in nuclear extract was detected by electrophoretic mobility shift assay (EMSA). RESULTS: All of the three anti-TNF mAbs could inhibit TNF-induced NF-kappaB nuclear translocation in a dose-dependent manner. At the concentrations of 10 mg/L and 0.1 mg/L, the inhibition rates of mAb D2, E6 and F6 were 94.2% and 75.1%, 64.9% and 28.6%, 70.3% and 49.5% respectively, while the inhibition rate of control mAb was only 20.0% and 11.1%. CONCLUSION: mAbs D2, E6 and F6 can specifically inhibit TNF-induced NF-kappaB nuclear translocation, which lays the foundation for preparation of therapeutic chimeric anti-human TNF antibody for treatment of infectious and autoimmune diseases.