External Quality Assessment of Syphilis Serological Tests in Shaanxi / 现代检验医学杂志

Objective To understand the quality and test ability of syphilis serological tests among the laboratories of seconda-ry and higher medical institutions in Shaanxi province,in order to reinforce the quality control and the management of vene-real laboratory and improve the technical capability of them.Methods Five quality control samples,QC manual and reports were delivered,and detected for treponemal qualitative tests and non-treponemal qualitative and quantitative tests,respective-ly.Syphilis laboratories were requested to provide feedback on the test results and other information within the specified time for a final statistical analysis.Results 341 laboratories participated in this assessment,the total qualification rates was 70. 97%.The coincidence rate of non-treponemal qualitative and quantitative tests were 97.69% and 76.16%,respectively.The coincidence rate of treponemal qualitative test was 9 9.9 1%.Conclusion The syphilis serological testing capacity of laborato-ries in Shaanxi province should be improved,the coincidence rate of non-treponemal quantitative tests was low.A program of improving external quality assessment of syphilis serological testing among different laboratories should be established and the professional training and the management system should be strengthened.

IL-1βstimulated neuron activation via PI3K/Akt/mTOR pathway / 中国病理生理杂志

[ ABSTRACT] AIM:To study the effect of interleukin-1β( IL-1β) on neuron activation during the process of me-dial temporal lobe epilepsy ( MTLE ) .METHODS: IL-1β, rapamycin [ an inhibitor of mammalian target of rapamycin (mTOR)]and lentiviral transfection to knockdown PI3K-p85 were used to pre-treat the neurons.The protein levels of PI3K-p85, p-Akt, p-p70S6K and MAP2 were detected and the relationship among the tested cytokines was analyzed.The neuron endocytosis was observed in each group.RESULTS:IL-1βincreased the protein levels of PI3K-p85, p-Akt and p-p70S6K, up-regulated the expression of PI3K-p85 binding with IL-1RI in the neurons, and increased the neuron endocyto-sis compared with control group (P<0.05) .These processes were inhibited by rapamycin and silence of PI3K-p85 (P<0.05).Inhibition of the PI3K-p85 binding to IL-1RI decreased the protein levels of p-Akt, p-p70S6K and MAP2 which were increased by IL-1βstimulation (P<0.05).CONCLUSION: IL-1βactivates PI3K-p85 by binding with IL-1RI to promote the activation and proliferation of neuron synapses via PI3K/Akt/mTOR signaling pathway, which might be one of the mechanisms in MTLE chronic progress.

Studies on quality evaluation of Dendrobii devoniani / 中国中药杂志

To establish the local quality standard for Dendrobii devoniani caulis from Longling, Yunnan, the pharmacognostic characteristics microscopic characteristics and TLC identification were developed. Sulfuric acid-phenol method was used to determine the content of polysaccharide. An HPLC method was adopted to determine the content of mannose, and extractives were determined according to the procedures recorded in the Appendix of Chinese Pharmacopoeia(2010). The results showed a strong characteristics microscopic of Dendrobii devoniani caulis, and its TLC identification had a good resolution with clear spots; the content of polysaccharide is 35.7% -52.1% (average 42.7%), mannose 27.8%-46.1% (average 35.8%), and extract 4.5%-10.6% (average 7.38%). The method is simple, accurate and reliable, with good reproducibility. The established standard is acceptable for quality evaluation of Dendrobii devoniani caulis from Longling, Yunnan.

Clinicopathological analysis of 141 pediatric autopsy cases / 中华儿科杂志

<p><b>OBJECTIVE</b>To summarize the major pathological findings, causes of deaths and reasons for misdiagnosis of 141 autopsy cases and thereby to improve the diagnosis level and reduce misdiagnosis.</p><p><b>METHOD</b>A retrospective analysis of pathological reports and clinical materials of 141 pediatric autopsy cases from June, 1986 to June, 2006 of our department was performed. Classification was based on (1) international classification of diseases of the World Health Organization; (2) age: cases 28 d-3 years old were defined as infants and young children group, -7 yeas olds were defined as preschool age group, -14 years olds were school age group; (3) when statistics was conducted, the first 3 items of the clinical diagnoses were counted. If one of them was consistent with the pathological diagnosis, it was regarded as basically in accordance with the pathology, if none of the first 3 was consistent with pathological diagnosis, the case was regarded as misdiagnosed.</p><p><b>RESULTS</b>(1) The top three major pathological diagnosis and causes of death were: 1) Classified according to system: 41 cases had tumor (29.1%), 25 cases had respiratory diseases (17.7%), 18 cases had infectious diseases (12.7%); 2) Classified according to disease: 18 cases had malignant histiocytosis, 12 cases had sepsis, 11 cases had lobular pneumonia. (2) The causes of deaths changed gradually. The top cause of death was respiratory diseases during the former 10 years and was tumor during the latter 10 years; the materials showed that 95 cases were 28 d-3 years old (67.4%), and some rare diseases, such as mediastinal and lung chorionic epithelioma (choriocarcinoma), and pulmonary alveolar proteinosis were found. (3) In 90 cases the clinical diagnosis was basically in accordance with the pathological diagnosis (63.8%) and misdiagnosis was found in 51 cases (36.2%).</p><p><b>CONCLUSION</b>For clinical diagnosis of critically ill patients, both common and rare diseases should be considered. Analysis of autopsy materials could confirm and/or correct clinical diagnosis and is helpful to summarize clinical diagnosis experience.</p>

The tight junction proteins ZO-1, occludin and actin participate in the permeability increasing of blood-brain barrier induced by hypoxia-ischemia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the changes of blood-brain barrier-tight junction (BBB-TJ) proteins ZO-1, occludin and actin following hypoxia-ischemia (HI) in order to explore the possible mechanism of permeability increasing of blood-brain barrier (BBB) induced by HI.</p><p><b>METHODS</b>BBB models were established by co-culture of cell ECV304 and astrocytes (AS) in vitro, then randomly assigned to control and HI groups. Transmission electron microscope was used to observe the changes of BBB-TJ. The distribution of actin was determined by direct-immunofluorescence microscope. Definite permeability of BBB models by 125I-BSA was detected by gamma events-per-unit-time meter. Expression of actin, ZO-1 and occludin was detected by Western blot.</p><p><b>RESULTS</b>After 10-day culture, endothelial cells connected tightly, with plenty of TJ which was smooth, continuous and of high density, in the BBB models. After 5 hrs of HI, the TJ was opened with intercellular gaps formation. The direct immunofluorescence showed that the peripheral filament bands became blurred, the cell-cell junction loosened and fissure appeared in the HI group. The permeability of 125I-BSA in the HI group increased significantly compared with the control group (P<0.01). Expression of ZO-1 decreased markedly, while expression of actin and occludin was not different in the HI group compared with the control group.</p><p><b>CONCLUSIONS</b>The changes in occludin distribution and decreased expression of ZO-1 lead the reorganization of BBB-actin protein, which may be one of the mechanisms of permeability increasing of BBB following HI.</p>

A pilot study on the proteome of cerebrospinal fluid of Staphylococcus epidermidis meningitis in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Bacterial meningitis is a kind of central nervous system infection with a high incidence, disability and fatality in children. Prompt diagnosis and treatment are associated with an improved prognosis. Low positive rate of bacterial cultures of the cerebrospinal fluid (CSF) makes it difficult to make a definite diagnosis. This experiment aimed to investigate a proteome profile of normal CSF of Chinese children by two-dimensional polyacrydamide gel electrophoresis (2-DE), and to sieve the disease-specific proteins of Staphylococcus epidermidis meningitis (SeM) to provide basis for early diagnosis and treatment of SeM.</p><p><b>METHODS</b>Four mL CSF samples were obtained respectively from SeM and normal children. The separated proteins with immobile pH gradient (IPG) 2-DE technology and protein spots were visualized by Coomassie Brilliant Blue staining. The stained 2-DE gels were scanned on the Imagescanner and pictures were obtained through Labscan software. The images were analyzed with PDQuest software and the differences of protein spots were compared between the SeM and normal children.</p><p><b>RESULTS</b>Mean protein spots of the 2-DE gels were 438 and 425 in the SeM and normal groups respectively. Twenty-five protein spots only occurred in normal CSF and 12 spots only occurred in the SeM group. The expression of 6 protein spots showed up-regulation and that of 19 showed down-regulation in the SeM group compared with that in the normal group.</p><p><b>CONCLUSIONS</b>A 2-DE profile of CSF proteome was successfully established in SeM and normal children through proteomic technique. By the differentiated analysis of these CSF 2-DE gels, the differences of CSF proteome profiles were found between SeM and normal children. Future analysis and identification of these spots will contribute to find out the disease specific proteins of SeM and to provide basis for early diagnosis and therapy of this disorder.</p>

Tumor necrosis factor-alpha increases in vitro blood brain barrier permeability / 中国组织工程研究

BACKGROUND:The levels of interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) are increased during infectious brain edema, and are positively relevant to the degree of brain damage. However, whether TNF-α can enhance blood brain barrier (BBB) permeability remains unclear, especially in vitro.OBJECTIVE: To understand the changes and possible mechanism of the BBB permeability induced by TNF-α in vitro.DESIGN: Randomized controlled cell model study in vitro.SETTING:Department of Pediatrics, Xiangya Hospital, Central South University; Department of Biochemistry, Xiangya Medical College, Central South University.MATERIALS: Twenty 7-day-old healthy Sprague-Dawley rats, of clean grade and either gender, were provided by the Animal Center, Xiangya Hospital, Central South University. TNF-α was purchased from sigma Company; DMEM fluid medium and fetal bovine serum were purchased from Hyclone Company; Y-27632 was purchased from Alexis Company,and rabbit anti-human factor Ⅷ -related antigen was purchased from Zymed Company; Mouse anti-rat glial fibrillary acidic protein (GFAP) was purchased from Neomarkers. Other biochemical reagents were imported (Sigma Company).METHODS: This experiment was carried out in the Xiangya Hospital, Central South University between March 2004 and April 2005. Brain microvascular endothelial cells and astrocytes were co-cultured 10 days to set up rat models of BBB in vitro. Then, the cells were divided into 4 groups: model group(BBB models were prepared), TNF-α group ( BBB model incubated with 0.01 g/L TNF-α for 5 hours), Y-27632 pretreated group ( BBB model incubated with 30 μmol/L Y-27632 for 1 hour before 0.01g/L TNF-α challenge ) and Y-27632 control group (BBB models only incubated with Y-27632 as those in the Y-27632 pretreated group). The effect of TNF-α on BBB permeability was observed by detecting the 125 I -BSA, which passed through the inserts at each time point (30,60,120 and 240 minutes) using .γradioimmunoassay counter.MAIN OUTCOME MEASURES: The 125 I -BSA, which passed through the inserts in rat models of BBB at different time points after intervention.RESULTS: The 125 I -BSA, which passed through the inserts in rat models of BBB, was all significantly higher in the TNF-α group than in the other groups at 30, 60, 120 and 240 minutes after intervention, respectively (P ＜ 0.01), and reached the peak at 240 minutes; The 125 I -BSA, which passed through the inserts, was lower in the Y-27632 pre-treated group than in the TNF-α group at 30 and 60 minutes after intervention (P＜ 0.01). There was also significant difference in 125 I -BSA permeation between Y-27632 pretreated group and Y-27632 control group after 120 minutes (P ＜ 0.05).CONCLUSION: TNF-α can increase BBB permeability, and Y-27632 pretreatment can early reverse the effect of TNF-α on BBB permeability.

Matrix metalloproteinase-9 and cystoskeleton actin are involved in the increased permeability induced by hypoxia/ischemia status in a blood-brain barrier model / 中国病理生理杂志

AIM:To understand the effects and approach the mechanisms of matrix metalloproteinase-9(MMP-9)and cystoskeleton actin on the permeability increasing of blood-brain barrier(BBB)model which was induced by hypoxia/ischemia status in vitro.METHODS:The BBB model was build by the co-culture of cell ECV304 and astrocytes in vitro,then divided randomly into control group,hypoxia/ischemia group and BB-1101 pretreatment group.The permeability of BBB was determined by [125I]-BSA.The expression and the disposition of actin were detected by direct-immunofluorescence and Western blotting.BB-1101,the MMPs inhibitor,was used to investigate if MMP-9 participate the process of the increasing of BBB models' permeability in hypoxia/ischemia status.RESULTS:Post-stimulation of hypoxia/ischemia for 5 h,the permeability of [125I]-BSA and amount expression of MMP-9 in hypoxia-ischemia group was increased compared with control group(P

Influence of Tumor Necrosis Factor-? on Blood Brain Barrier Permeability and Its Mechanism / 实用儿科临床杂志

Objective To understand the changes and possible mechanism of the blood brain barrier(BBB) permeability induced by tumor necrosis factor(TNF-?) in vitro.Methods BBB model was established by coculturing allogenic brain microvessel end othelial cell(BMEC) and astrocyte(AS).The BBB model was divided randomly into normal control group,TNF-? group and Y-27632 pretreatment group.The changes of BBB permeability were evaluated by Gamma radioim muno assay counter.Results The Gamma radioimmuno assay indicated that the marker,~(125)I-BSA,across the BBB model in vitro was significantly increased after TNF-? treatment compared with control group,Y-27632 pretreatmen could prevent the permeability of BBB induced by TNF-?(P

Lysophosphatidic acid (LPA) induces the proliferation of astrocytes in rat via pathway of protein kinase C-? and calcium ion in vitro / 中国病理生理杂志

AIM:To determine if lysophosphatidic acid(LPA)regulates the proliferation of astrocytes(AS)and to approach the mechanism of the process.METHODS:The cerebral AS of the neonatal SD rats were cultured in vitro and divided randomly into control group,PKC excitomotor(PMA)group,LPA group,PKC-? inhibitor(Ro31-8220)group,Ro31-8220+PMA group and Ro31-8220+LPA group.The proliferation of the cells was detected by MTT assay and flow cytometry(FCM).The concentration of intra-cellular calcium ion of the cells([Ca~(2+)]_i)which were labeled with Fura-2/AM was determined by ultraviolet spectrophotometer.The change of PKC-? inside the cells was observed by Western blotting.RESULTS:LPA and PMA stimulated the proliferation of AS,they also enhanced the expression of PKC-? and increased the concentration of [Ca~(2+)]_i.After pretreated with Ro31-8220,the abilities of LPA that mentioned above were decreased.The change of [Ca~(2+)]_i was associated with the diversity of PKC-?.CONCLUSION:LPA promotes the proliferation of AS via the way of PKC-? and Ca2+.