OBJECTIVETo observe the protective effect of active fractions of Huanglian Jiedu Decoction (HJD) on primary cortical neuron injury after oxygen-glucose deprivation (OGD)/reperfusion (R) injury. Methods Using macroporous resin method, HJDFE30, HJDFE50, HJDFE75, and HJDFE95 with 30%, 50%, 75%, and 95% alcohol were respectively prepared. Then the content of active components in different HJD fractions was determined with reverse phase high-performance liquid chromatography (RP-HPLC). The OGD/R injury model was induced by sodium dithionite on primary cortical neurons in neonate rats. MTT assay was used to observe the effect of four fractions (HJDFE30, HJDFE50, HJDFE75, and HJDFE95) and seven index components of HJD on the neuron viability.

RESULTSRP-HPLC showed active component(s) contained in HJDFE30 was geniposide; baicalin, palmatine, berberine, and wogonside contained in HJDFE50; baicalin, berberine, baicalein, and wogonin contained in HJDFE75. The neuron viability was decreased after OGD for 20 min and reperfusion for 1 h, (P <0. 01), and significantly increased after administered with HJD, HJDFE30, HJDFE50, and HJDFE75 (P <0. 05, P <0. 01). Geniposide, baicalin, baicalein, palmatine, wogonside, and wogonin could increase the cortical neuron viability (P <0. 05, P <0. 01).

CONCLUSIONSHJDFE30, HJDFE50, and HJDFE75, as active fractions of HJD, had protective effect on primary cortical neuron injury after OGD/R. Furthermore, geniposide, baicalin, and baicalein were main active components of HJD.

Animals ; Berberine ; Berberine Alkaloids ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Flavanones ; Flavonoids ; Glucose ; metabolism ; Iridoids ; Models, Animal ; Neurons ; Oxygen ; metabolism ; Rats ; Reperfusion Injury ; drug therapy

Objective To test the reliability and validity of CRIES-13(Chinese edition). The Children Revised Impact of Event Scale (CRIES-13) was recommened for diagnosing PTSD of children. Methods In the last third of the September 2008,according to the suffering condition,600 students were choosed who were fit for the research standard as subjects in two middle schools randomly. The viability of CRIES-13 was weighted by testretest reliability,Cronbach' s alpha,Split half reliability. The validity of CRIES-13 was analysed by content validity ,criterion validity,construction validity. Results In the test-retest reliability of CRIES-13, the Spearman correlation coefficient of total,intrusion factor,avoid factor,high warkening factor were 0.79, 0.75, 0.71, 0.75. Significant correlation were found among these scores. The Cronbach' s alpha of population was 0. 81. The Cronbach' s alpha of three factors was 0. 79 ( intrusion factor) , 0. 71 ( avoid factor), 0. 65 ( high awarkening factor). CRIES-13's split-half reliability was 0. 85. In the content validity test,the Spearman rank correlation coefficient between total score and each item was 0. 83 (intrusion factor), 0. 75 (avoid factor), 0. 85 (high awarkening factor). The correlation between intrusion factor and avoid factor was 0.63. The correlation between avoid factor and high awarkening factor was 0.41. The correlation between intrusion factor and high awarkening factor was 0.41. In struction validity, variance orthogonal rotation factor analysis was adopted and got three general factors. Their cumulative contribution to total variance was 55.52%. In the criterion validity test,significant correlation was found between intrusion factor and SDQ emotional factor and depression scale total score. Significant correlation was found between high awakening factor and SDQ emotional factor and depression scale total score. Conclusion The reliability and validity of CRIES-13 was good. It could be used extensively.

Objective The Children's Revised Impact of Event Scale (CRIES-13) is used for Screening PTSD of children. The reliability and validity of CRIES-13 is good. To research the demarcation points of CRIES-13 (Chinese version) based on the reliability and validity analysis,and to improve the useful value of the scale.Methods In late September 2008, according affected condition, students were choosed who were fit for the research standard as subjects in two middle schools. First,general questionnaire (self-writing) and CRIES-13 were applied to the subjects. Second, according to K-SADS-PL, physician carried out diagnosis meeting and evaluation to 310 students who were classified by stratified rand sampling. Critical point of CRIES-13 was divided by K-SADSPL. The assessment value of it were sensitivity, specificity, veracity, PPV, NPV. The right choice of division was measured by ROC curve. Results When the critical score was higher than 30, the score of Se ( 0. 833 ), Sp(0. 836) and NPV (0.97) was in the high level. Conclusion When the critical score is higher than 30, the scale have a good discrimination for PTSD, non-PTSD and it can be used extensively.

Background Glial cells perform specialized function in many aspects of the development,homeostasis,and function of neurons.Retinal ganglion cells (RGCs)and glia interactions are critically important in glaucomatous neurodegeneration.However,the precise mechanisms of glial activation and ganglion cells damage are still remained unclear. Objective This study was to assess the early responses of glial cells in the retina,optic nerve and optic chiasm in rat models of acute high intraocular pressure (IOP),and to examine the expression of nestin,a neuronal progenitor marker,in the reactive glias. Methods Acute high IOP of 110 mmHg was induced in the right eyes of 6 clean adult female Wistar rats by infusing normal saline solution into the anterior chamber for 60 minutes.Three normal matched Wistar rats were used as controls.The rats were sacrificed by overanaesthesia and sections of retina,optic nerve and optic chiasm were collected on 3 days and 7 days after the injection.Rat retina was examined by Nissl staining to illustrate the gross structure changes.Loss of axons of RGCs in the optic nerve was assessed by immunostaining of β Ⅲ-tubulin.Double labeling of glia] fibrillary acidic protein (GFAP) and nestin was performed in sections of retina,optic nerve and optic chiasm to evaluate the glial responses.The use of the animals complied with Statement of Animal Ethic Committee of Peking University Third Hospital. Results In control rats,GFAP-positive glial cells were observed in the retina,optic nerve and optic chiasm,where only weak positive response for nestin was noticed.Three days after acute IOP elevation,thickness of inner plexus form layer was significantlydecreased in comparison with the control rats.A loss of 46％ RGCs was found in the rats with ocular hypertension.Obvious increase of GFAP expression was displayed in the retina,and processes of GFAP-positive glia cells extended into outer retina accompanied with significant up regulation of nestin.Axons in the optic nerve demonstrated a tendency of degeneration.Nestin expression increased significantly in the GFAP-positive glias in the optic nerve.Cross-sectional area of optic chiasm corresponding to the injured retina decreased relative to its countcrpart.Astrocyte like GFAP and nestin-colabeled glials were observed in this part of optic chiasm.The pathological changes of the retina,optic nerve and optic chiasm in hypertensive eyes aggravated on 7 days. Conclusions Acute ocular hypertension induce early onset of RGCs loss and axon degeneration.Neuronal injury is accompanied with glial reaction.Reactive glial cells express neuronal progenitor markers.The structural changes of the optic nerve and optic chiasm occur simultaneously with the high IOP.

ObjectiveTo evaluate the nephroprotective effects of transplanting metanephric mesenchymal cells (MMCs) into the renal subcaspsule of rats with acute tubular necrosis (ATN) induced by gentamicin. MethodsMMCs were expanded in culture and immunocytochemistry was used to characterize the cells. After gentamicin-induced ATN, fluorescence-labeled cells were transplanted and traced in kidney tissues by fluorescence microscopy. Serum creatinine (Scr) and N-acetyl-b-D-glucosaminidase (NAG) were tested. Kidney pathology was studied by hematoxylin-eosin staining. Apoptosis was examined by the TUNEL assay. Ki-67 and Bcl-2 expression was examined by immunohistochemistry. ResultsMMCs were expanded in culture and the phenotype of the cells was vimentin-positive and keratin-negative. Compared with other ATN groups, in the MMCs-treated group, Scr and NAG clearly decreased[14d Scr: (101.38±20.46) μmol/L vs (248.78±23.15), (252.98±33.52), (229.08±18.18) μmol/L;NAG: (14.83±7.74) U/L vs (33.33±14.88), (29.62±10.54), (30.22±10.94) U/L, P<0.05, respectively];the histopathoiogic lesion scores were lower (P<0.05);the Ki-67 antibody and apoptosis of renal tubular epithelial cells were improved or reduced respectively;the expression of Bcl-2 protein was up-regulated (P<0.05). ConclusionThe subcapsular transplantation of MMCs can ameliorate renal function and repair kidney injury.

This paper summarizes the current literature on the potential therapeutic role of stem cell transplantation for kidney injury and repair and focuses on the choice of types of stem cells, the method of transplantation, and the mechanisms of stem cell homing to injured renal tissues and its protective effects. The application of umbilical cord mesenchymal stem cells (UC-MSCs) shows wide prospects, but the approach and optimal dose of cell transplantation are under intensive investigation. Signals that regulate stem cell homing to injured renal tissues may be related to chemokines or factors released in the target site. Several studies have pointed out that paracrine and endocrine of stem cells are the most likely mechanism of action in the injured nephron. Many questions remain unanswered but stem cell-based therapy is a promising new strategy for acute and chronic kidney diseases.
Animals ; Cord Blood Stem Cell Transplantation ; Humans ; Kidney Diseases ; therapy ; Mesenchymal Stem Cell Transplantation ; Stem Cell Transplantation ; methods

Objective To examine the expressions of FHIT gene and HER_2 in non-small cell lung cancer(NSCLC)and the relationship between the expressions and the clinicopathological features of NSCLC, and to investigate the relationship between FHIT protein and HER_2 expression. Methods FHIT protein and HER_2 expressions in 50 lung cancer cases and 21 adjacent non-cancer tissues were performed by immunohistochemistry. And the positive rates of FHIT and HER_2 were measured. Results The positive rates of FHIT protein were 30.00% in lung cancer tissues. The expression levels of FHIT were significantly lower in lung cancer tissues than that in non-cancer lung tissues (P

0.05)in healthy volunteers.

Objective To do screening acute lymphoblastic leukemia patients scFv antibody single chain variable region to cre-ate conditions for the expression and obtain further specificity of antibody fragments.Methods In this study,patients with newly diagnosed acute lymphoblastic leukemia serum as coating antigen using phage display technology,screening phage an-tibody specificity from the semi-synthetic human phage antibody libraries,the first to target the immune antigen-coated tab-let,phage library was added,so that with the target antigen-specific binding phage antibody was immobilized on plates immu-nization,could not be specifically bound phages were rinsed.The eluted specific binding phage,E.coli infection.Could get the specific antibody gene containing phagemid.Results After three “adsorption-elution-amplification”screening process,got stronger leukemia patient antigen-specific phage antibody variable region fragment and identification.Conclusion Got better strain affinity antibody fragments,to create the conditions for the next fragment expression,identification and clinical appli-cation.

Objective To establish a time resolved fluoroimmunoassay (TRFIA) method for detecting Glypican 3 (GPC3) and to explore the diagnostic value of serum GPC3 for hepatic carcinoma (HCC). Methods Microplate coated with anti-GPC3 monoclonal antibody 7C8 and GP9 labeled with Eu3+ were used to establish TRFIA kit. The serum concentrations of GPC3 in 41 HCC patients and 44 chronic hepatitis (CH) patients were quantitatively analyzed. AFP was detected by with lowest limit of 2.06 μg/L. The CV of inter and intra assay were 12.25% and 12.91%, respectively. The average serum concentration of GPC3 in HCC patients was (86.68±110.39) μg/L (median: 56.98 μg/L). But in CH patients it was only (14.77±29.48) μg/L, which was significantly lower than that in HCC (Wilcoxon W=1335.00, Z=-4.99, P<0.001). With diagnostic cut-off value set at 42.94 μg/L, the diagnostic sensitivity and specificity of TRFIA GPC3 for HCC were 58.5% (24/41) and 95.5%(42/44) respectively. The diagnostic sensitivity of AFP was 46.3% (19/41) in 41 HCC patients, and was raised to 78.0% (32/41) when combined with GPC3. Conclusions Serum GPC3 assay by TRFIA is established and it could increase the diagnostic sensitivity for HCC when combined with AFP.