Decision Making, Organizational ; Disasters ; Earthquakes ; Humans ; Public Health Practice ; Stress Disorders, Post-Traumatic

Objective To compare the distribution and expression differences of glycogen synthase kinase-3?(GSK3?) among normal adult,aged and amyloid beta(A?)-induced neurodegenerative rat brains,so as to explore its functional role in neurodegeneration. Methods Aggregated A? was microinjected into normal adult rat hippocampus under a stereotaxic system. The rats over 12 months were defined as aged rats. The distribution and localization of GSK3? were examined using immunohistochemistry. Western blotting was performed to assess expression change in cortex and hippocampus quantitatively. Results The GSK3? positive cells were distributed extensively around the whole brain and almost with neuron-like morphology. In normal adult rats,the strong anti-GSK3? immunoreactivity located in the neocortex pyramidal layer,hippocampus pyramidal layer,dentate gyrus,thalamus,substantia nigra,etc. The amount of GSK3? positive cells was much more in the aged and A?-injected group than in normal ones. The immunoreactive signals usually extend to the distal area of neurite in the A?-injected ones. Western blot showed that the expression intensity of GSK3? was stronger in the aged and neurodegenerative rat brain than in the normal adult rat brain. Conclusion The expression of GSK3? increases apparently in the neurons of aged and A?-injected brain. It may play a role in the neurodegenerative process.

OBJECTIVETo introduce a new measuring tool for measuring postoperative limb length exactly, and to provide a convenient and effective method to control limb length after total hip replacement.

METHODSFrom January 2013 to September 2014, 102 patients undergoing primary unilateral hip replacement were divided into two groups: experimental group and control group. There were 51 patients in the experimental group, including 25 males and 26 females, ranging in age from 37 to 92 years old, with an average of 60.41 years old. The patients in experimental group were treated with new method to control limb length. Other 51 patients in the control group, including 27 males and 24 females, ranging in age from 35 to 87 years old, with an average of 61.00 years old. The patients in the control group were treated with normal methods such as shuck test or limb touching. All the patients were operated by the same experienced surgeon. In the experimental group,total hip arthroplasties (THA) were performed on 35 patients with avascular necrosis of the femoral head or femoral neck fracture, and 16 patients were treated with hemiarthroplasty (HA). In the control group, 38 patients received THA and 13 patients received HA. On the anterior-posterior X-ray radiograph, several indexes were measured as follows: the distance of bilateral femoral offset (a), the height from tip of great trochanter to the rotation center of the femoral head (b) and the vertical distance between the top of the minor trochanter and the two tear drops line (c). The leg length discrepancy can be assessed with three parameters as follows: d1, the absolute value of the difference between the bilateral a values; d2, the difference between the bilateral b values; d3, the difference between the bilateral c values. The SPSS 21.0 was applied for the statistical analysis.

RESULTSIn the experimental and control groups, d1 were 4.49 mm and 7.32 mm (P = 0.013); d2 were 2.37 mm and 4.32 mm (P = 0.033); d3 were 3.32 mm and 6.08 mm (P = 0.031). The values of d1, d2 and d3 in the experimental group were significant smaller than those in the control group.

CONCLUSIONThe new measuring tool and method can be used to control the limb length and offset effectively during operation.

Adult ; Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; adverse effects ; methods ; Case-Control Studies ; Female ; Humans ; Leg Length Inequality ; prevention & control ; Male ; Middle Aged

Objective To investigate the effect of lithium chloride(LiCl),an inhibitor of glycogen synthase kinase-3beta(GSK-3beta),on proliferation and differentiation of neural stem cells(NSCs).Methods The NSCs were isolated from cortex of rat fetus and expanded in culturing system.Their morphological changes and attachment process were observed under microscope.The cell cycle dynamics of NSCs was examined with flow cytometry.And the expression of GSK-3β and β-catenin was examined quantitatively with Western blot.Results The culturing NSCs treated with LiCl were usually floated and much dispersed in the media.Many of the neurospheres became small and the time of attachment after serum induction became longer.Using flow cytometry,it was detected that the proportion of G1 phase NSCs declined gradually accompanying the increased concentration of LiCl,while the percentage of S and G2/M phase cells showed an increasing trend.Western blotting results revealed β-catenin expression increased whereas Gsk-3βdecreased gradually under the treatment of LiCl and also showed a dose dependent manner.Conclusion These results suggest that LiCl may promote the proliferation of NSCs and prevent them from differentiating,which may partly involve the activation of wnt/β-catenin signaling pathway.

AIM: To investigate the effect of human amniotic fluid on the release of thromboxane A 2 (TXA 2), prostaglandin I 2 (PGI 2) and Leukotriene C 4(LTC 4) from blood cells. METHODS: 1 mL human amniotic fluid and 10 mL oneself blood collected from 38-41 weeks with cesarean section were cultured at 37℃ for 30 min, and then centrifuged. The supernatants were taken and stored at -70℃. TXB 2 and 6-Keto-PGF 1? of the superntants were determined by radioimmunoassay and LTC 4 by enzyme immunoassay. RESULTS: It was found that the levels of TXB 2 and LTC 4 in blood were elevated from (63.5?52.0) ng/L and (40.1?39.2) ng/L to (189.1?102.0) ng/L and (293.5?206.1) ng/L respectively (P0.05).CONCLUSION: Amniotic fluid might stimulate the release of TXA 2 and LTC 4 from blood, it might affect the balance of TXA 2 and PGI 2 in blood, which might play an important role in the pathogenesis of amniotic fluid embolism.

Objective To establish a new assay for detecting autoantibody Jo-1, cloning and expressing human autoantigen Jo-1 in E.coli.Methods A full length cDNA of human autoantigen Jo-1 was cloned from cell line HL-60 by RT-PCR. The PCR product was TA cloned, sequenced and inserted into the carrier pGEX-5T.The recombinant plasmid was transformed into E.coli BL-21. The positive clones were identified by restricted enzymes and induced by IPTG. The expression product was analyzed by SDS-PAGE and Western blot.Results The PCR product was about 1 500 bp in size which was in accordance with predicted 1 526 bp and sequencing result showed the same with GenBank′s report.The pGEX-5T- Jo-1 positive clone produced a 75 000 fusion protein which had natural immunogenicity of human autoantigen Jo-1 by SDS-PAGE and Western blot.Conclusion Successfully cloning and expression of human autoantigen Jo-1 laid a foundation for further research work.

Objective To clone,express and identify the nuclear antigen Sm B′in E. coli to establish a new assay for detecting autoanti-body to Sm B′. Methods A full length cDNA of Sm B′was cloned from cell line HL-60 by RT-PCR. The PCR product was TA cloned and sequenced and inserted into the vector pGEX-5T. The recombinant plasmid was transformed into E. coli BL21. The positive clones were identified by restricted enzymes and induced by IPTG. The expression product was analyzed by SDS-PAGE and Western blot. Results The PCR product was about 700 bp in size which was in accordance with predicted 657 bp and sequencing result showed consistent with the sequence in GenBank. The pGEX-5T-Sm B′positive clone produced a 51 000 kD of fusion protein which was immunoreac-tive with anti-Sin B′confirmed by SDS-PAGE and Western blot. Conclusion The successful cloning and expression of nuclear antigen Sm B′laid a foundation for further research work.

BACKGROUND: Occurrence and development of the glioma are not only related to regulative gene of cell cycle, but also dealt with adjustive genes of cell invasion, cell metastasis and apoptosis.OBJECTIVE: To identify the differentially expressed genes related to cell proliferation and apoptosis in glioma cell of different differentiating degree with cDNA microarray, and provide basic data for further research of mechanisms of cell proliferation and apoptosis in human gliomas.DESIGN: Opening-up experiment.SETTING: Institute of Pathology, Southwest Hospital and Department of Neurobiology, Basic Medicine Faculty as well as Department of Pharmacy,Xinqiao Hospital, Third Military Medical University of Chinese PLA.MATERIALS: Human malignant glioma cell line CHG-5 ( Ⅱ grading according to the WHO standard) was constructed, kept and cultured in this experiment. SHG-44 (Ⅳ grade according to the WHO standard) was provided by Institute of Brain Tumor, Second Hospital Affiliated to Suzhou Medical College. Serum of calf was produced and provided by Hangzhou Sijiqing Biomaterial Institute. In experiment RPMI-1640 medium (Gibco),Trizol test kit (Gibco-BRL), RNAsecureTM solutions (Ambion, Austin,Texas), biophotometer (Eppendorf, Hamburg, Germany) were also applied.Gene chip contained 9 984 human cDNA segment, which was prepared and provided by City University of Hongkong (UniGEMV2 cloneset known gene and ESTs were purchased from Incyte Company), Superscript Ⅱ reverse transcriptase was provided by Gibco-BRL Company. Fluorochrome Cy3 & Cy5 was the products of Amersham Pharmacia Company.METHODS: The experiment was performed at Chongqing Third Military Medical University of Chinese PLA and City University of Hongkong between 2001 and 2003. Total RNA was extracted from Trizol test kit. Reverse transcription polymerase chain reaction (RT-PCR) was conducted in Superscript Ⅱ reverse transcriptase, and cDNA product was marked with fluorochrome Cy3 & CyS. Followed by chip hybridizationto detect the difference of gene expressions between human glioma cell line CHG-5 and SHG-44 tumor cell, especially the difference of related genic expression between cell proliferation and apoptosis, and the chip result was verified with Northern blot hybridization.MAIN OUTCOME MEASURES: The difference of gene expression of human glioma cell in different differentiating degree; Comparison of result between Northern blot and related gene chip.RESULTS: ①Compared with CHG-5, 120 gene expressions were detected obvious up-regulation and 22 gene expressions were significant down regulation in SHG-44 cells, and the variety of these differentially expressed genes was many, in which apoptosis related genes were 6, including 3 up-regulation genes and 3 down-regulation genes; Cell cycle and proliferating related genes were 12, including 5 up-regulation genes and 7 down-regulation genes. ②Chip result was supported by Northern blot result further.CONCLUSION: The differentially expressed genes of glioma are revealed primarily, especially the differentially expressed genes related with cell proliferation and apoptosis.

AIM: To evaluate the therapeutic efficacy of intratracheal instillation of porcine pulmonary surfactant(PPS) in rats with lipopolysaccharides(LPS)-induced early-stage ALI in this study.METHODS: SD rats weighing 200 g-300 g were randomly divided into 4 groups: LPS(1.5 mg?kg-1)+saline,LPS+PPS 100 mg?kg-1,LPS+PPS 150 mg?kg-1,LPS+PPS 200 mg?kg-1.The PaO2 and PaCO2,as well as survival rate of rats were examined for 6 h after the start of PPS-instillation.Then,rats were killed and lungs were immediately removed for lung index(LI) and histological analysis.The bronchoalveolar lavage fluid(BALF) was collected for measurement of total protein(TP) contents,TNF-? level and white blood cell(WBC) numbers.RESULTS: Significantly increased PaO2,reduced mortality rate,decreased total protein and TNF-? contents in BAL,as well as lung index and meliorated histological appearance were observed in three PPS-treated groups compared with group given saline after LPS(P

Objective To study the protective effect of fluvastatin,one of the HMG-CoA reductase inhibitors (statins),against oxygen radical-induced oxidative damages in human aortic endothelial cell,and the role of Bcl-2 in this protection.Methods Human aortic endothelial cells with or without Bcl-2 siRNA transfection were subjected to 1-100 nM of fluvastatin and 100 la hydrogen peroxide for 24 hours.Bcl-2 mRNA and protein expression were measured by Taqman quantitative PCR and Western blotting.Cell apoptosis was measured by normal and fluorescent microscopy and Cell Death Detection ELISA.Results In the Bcl-2-expressed cells,fluvastatin significantly reversed hydrogen peroxide-induced microscopic apoptosis and apoptotic DNA fragmentation,which were accompanied by a markedly upregulation of Bcl-2 expression by fluvastatin.However,the endothelial protection by fluvastatin was completely lost in Bcl-2 siRNA transfected cells.Conclusion Fluvastatin protects human endothelial cells against oxygen radical-induced cell apoptosis in vitro,and this protection seemed to be mediated in a Bcl-2 dependent pathway.(J Geriatr Cardil 12008;5:33-38)