Objective To study the mutagenic action of antiepileptic drugs(AEDs), and find an effective way to prevent the mutagenesis induced by AEDs,by observing the effects of AEDs on serum folic acid(FA) level and sister chromatid exchange (SCE) frequency in epileptic children.Methods Ninty epileptic children were divided into different groups on the basis of the different drugs they had taken, then detected the two indexes at different time points.Results The serum FA level and SCE frequency of the patients significantly decreased and increased after they took carbamazepine (CBZ) and valproic acid (VPA)respectively. The two indexes went back respectively when supplied with FA.Conclusions Both CBZ and VPA possess mutagenic action, yet nitrazepam does not.FA may help repair the chromosome damage and reduce the mutagenesis effects.

Objective To investigate whether there is prethrombotic state in acute viral encephalitis.Methods Von willebrand factors (VWF),granule membrane protein-140(GMP-140) and D-dimer were measured in 60 children,30 cases of them had mild acute viral encephalitis, and other 30 cases of them had severe acute viral encephalitis.Results The figures mentioned above had marked difference in children with mild acute viral encephalitis during acute and recovery stage compared with controls.Conclusions Testing the expressive rate of GMP-140 on platelet surface,plasma VWF and D-dimer can find the prethrombotic state immediately. GMP-140,VWF and D-dimer in estimating the severe extent of disease and the prognosis play a major role. J Appl Clin Pediatr,2004,19(6):501-503

Objective Explore an approach with which stem cell can be used to cure traumatic spinal cord. Methods To better repair spinal cord injury, adult Sprague-Dawley (SD) rat MSCs was isolated and induced to differentiate into neuron-like cells with a Chinese medicine, Musk's polypeptide named Musk-1 in vitro and be seeded into microsurgical transectional model of adult rat spinal cord with cell differentiation and transplantation techniques. Results After cell transplantation, animals seeded with the rMSCs-derived neuron-like cells promoted significant improvement in function after spinal cord (P＜0.05,effective observation for 90 days ) when compared to the lesion-control group. Histology and immunocytochemical analysis confirmed a large number of rMSCs-derived neuron-like cells survived well in the transplantation zoon and numerous cells migrated within the host adjacent tissues as far as 6millimeters above and below the border of the lesion. Fluorescence gold retrograde tract tracing analysis revealed the positive motor neuron of fluorescence gold mark was found in the head side of rat spinal marrow ,corpus rubrum ofmesencephalic and corticospinal tract fibers. This suggested that corticospinal tract fibers of spinal cord area got regeneration and passed through the lession to reach the tail of spinal marrow.Conclusion These findings demonstrated that "pre-programmed" rMSCs can survive, migrate, integrate as well as restore spinal function and behavior in the microsurgical transectional model of rat spinal cord, and may serve as a suitable approach to traumatic spinal cord injury.

OBJECTIVETo investigate the changes in rat cardiac function and myocardium ultrastructure in response to ouabain treatment.

METHODSTwenty-four male SD rats were randomized into two equal groups to receive daily intraperitoneal injection of ouabain or saline for 4 consecutive weeks, and their systolic blood pressure (SBP) was recorded weekly. After 4 weeks of injection, echocardiography was performed and the hemodynamic parameters were measured by invasive cardiac catheterization, and the changes in myocardium ultrastructure observed using transmission electron microscopy.

RESULTSAfter 4 weeks of ouabain injection, no significant changes in the mean SBP occurred in comparison with the saline group, but echocardiographic examination showed significant increases in the left ventricular end-diastolic and end-systolic diameters, septum thickness, posterior wall thickness, left ventricular mass and isovolumetric relaxation time but significantly lowered E/A ratio, ejection fraction and fractional shortening after ouabain treatment (P<0.05). Invasive monitoring revealed significant attenuation of the left ventricular developed pressure, rate of pressure development (+dp/dt) and rate of pressure decay (-dp/dt), and increment of the left ventricular end diastolic pressure. Myofibrillar fragmentation, swelling of the cardiac myocytes, absence of the Z line, increases of the mitochondria and collagen fibers were found in ouabain group by transmission electron microscopy.

CONCLUSIONOuabain can induce left ventricular enlargement, cardiac wall thickening, myocardial ultrastructural alterations, systolic and diastolic dysfunction in rats before blood pressure elevation is detected, indicating that ouabain can directly cause cardiac damage in rats.

Animals ; Echocardiography ; Heart ; drug effects ; physiopathology ; Male ; Microscopy, Electron, Transmission ; Myocardium ; pathology ; ultrastructure ; Ouabain ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effects of shikonin on the proliferation, expression of CXCR4 and the migratory responses to CXCL12 in colorectal carcinoma cell line SW480.

METHODSThe proliferation of SW480 cells was assessed by MTT assay. Cell surface expression of CXCR4 was determined by flow cytometry. The migratory ability was determined by Transwell.

RESULTSShikonin inhibited the proliferation of SW480 cells in time- and concentration-dependent manner. The expression rate of CXCR4 in SW480 cells was 99.1%. After application of shikonin 0.01 micromol/L, 0.1 micromol/L and 1.0 micromol/L for 24 h, the expression rate of CXCR4 decreased to 76.0%, 59.1% and 35.5% respectively (F=1098.041, P <0.001), and the CXCL12-induced SW480 cell migratory inhibition rate was 25.2%, 38.5% and 55.7% respectively (F=48.970, P <0.001).

CONCLUSIONBesides having inhibiting tumor cell proliferation effect, Shikonin may also play a role in anti-metastasis via down-regulating the expression of CXCR4 and reducing the CXCL12-induced migratory response in colorectal carcinoma cell.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chemokine CXCL12 ; metabolism ; Down-Regulation ; Humans ; Naphthoquinones ; pharmacology ; Receptors, CXCR4 ; metabolism

Adult ; Arthroscopy ; methods ; Humans ; Male ; Scapula ; surgery ; Shoulder Joint ; surgery ; Young Adult

OBJECTIVETo clone human PD-1 gene, construct a prokaryotic expression plasmid and express in E. coli.

METHODSThe human PD-1 cDNA was cloned by RT-PCR from the total RNA, which was extracted from peripheral blood lymphocyte cell of the patient with chronic hepatitis B. Recombinant PD-1 protein was been expressed and purified after the prokaryotic expression plasmid had been constructed. It was identified by SDS-PAGE, DNA sequencing and amino acid sequencing.

RESULTSThe PD-1 gene was cloned and confirmed by DNA sequencing. The recombinant protein was expressed in E. coli. The purified protein was obtained, then been confirmed by amino acid sequencing.

CONCLUSIONThe human PD-1 gene was successfully cloned and expressed in E. coli, which lays the foundation for further study on the function and application of PD-1.

Amino Acid Sequence ; Antigens, CD ; biosynthesis ; chemistry ; genetics ; isolation & purification ; Apoptosis Regulatory Proteins ; biosynthesis ; chemistry ; genetics ; isolation & purification ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Genetic Vectors ; genetics ; metabolism ; Humans ; Polymerase Chain Reaction ; Programmed Cell Death 1 Receptor ; Prokaryotic Cells ; metabolism ; Sequence Alignment ; Sequence Analysis, DNA

Objective To investigate pathogenic bacteria distribution and drug resistance of the bacteria isolated from Chinese young and mid-age patients (14-65 years).Methods Collect and monitor the isolates from young and mid-aged patients (14-65 years) in 557 mem-ber hospitals of Grade three between January 1 and December 31 in 2012.Disk diffusion and automatic machine method are applied to anti-biotic resistance test.The data were analyzed and processed with software WHONET 5.6.Results There are 383443 isolates from young and mid-aged patients in 557 hospitals of Grade three.Gram-negative bac-teria is account for 69.33%, the major strains are Escherichia coli of 78915, Klebsiella pneumoniae of 48698, Pseudomonas aeruginosa of 36239 and Acinetobacter baumannii of 27858, respectively;Gram-posi-tive bacteria is account for 30.67%, Staphylococcus aureus of 33328, Staphylococcus epidermidis of 18818, Enterococcus faecalis of 13114 and Enterococcus faecium of 8517 are the top four.It was not found that Staphylococcus spp.were resistant to vancomycin and linezolid, and there were teicoplanin resistant oagulase -negative Staphylococcus account for 3.4%, Vancomycin resistant Enterococcus faecium 3.2%, Vancomycin resistant Enterococcus faecalis 1.5%.Frequen-cy of extended spectrum beta lactamase ( ESBLs) in Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis are separately 58.4%, 33.6%and 24.9%, which increase resistance to imipenem as 2.8%, 6.5%and 4.1%contrast to last year.Rate of carbapenem resistant to Acinetobacter baumannii was about 50%, only polymixin B has better ac-tivity 96.7%.Haemophilus were isolated less among the young and mid-aged patients, but were sensitive to most an-tibiotic.Conclusion Antibiotic resistant rate of young and mid-aged patients in China are more serious, especially increase of ESBLs producing Enterobacteriaceae, multi drug resistant non-fermenting bacteria and vancomycin resistant Enterococcus.Meanwhile, the resistant trend is various by time, so that it is important and tedious work to monitor the antibiotic resistance.

Objective Obtain the hepatitis C virus high purified subunit fusion protein and detect its immunogenicity.Methods With the vector of pET-11d,fusion protein was expressed in Escherichia coli BL21(DE3) after induced by IPTG.The protein was then purified by DEAE negative ion exchange chromatography and Ni2+ affinity chromatography.Western Blot analysis was used to detect the antigenicity of the fusion protein.At the same time,the sera were collected and prepared from the immunized experimental animals in order to investigate the immunogenicity of the protein by EIA.Results High purified hepatitis C virus subunit fusion protein was obtained successfully.The EIA indicated that the fusion protein could elicit specific antibodies in the animals with very high titers.Conclusion The hepatitis C virus subunit fusion protein expressed in prokaryotic system was proved to have strong immunogenicity.It could provide some helpful and useful information to the hepatitis C virus prophylactic and therapeutic vaccine development.

OBJECTIVETo study the effect of cyclooxygenase-2 (COX-2) on lymphangiogenesis in breast cancer.

METHODSBy the means of immunohistochemistry, COX-2, vascular endothelial growth factor-C (VEGF-C) and D2-40 were examined in the tissue samples of primary tumors from 94 patients underwent surgical resections for breast cancer from November 1998 to March 2002. Eighty-three patients were followed-up. The expressions of VEGF-C mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot in MDA-MB-231 cell lines by the treatment of selective COX-2 inhibitor Nimesulide at different doses. The expressions of VEGF-C protein were evaluated in MDA-MB-231 cells treated by PGE2 (1 microg/ml) and Trastuzumab (1 microg/ml), respectively.

RESULTSCOX-2 over-expression was observed in 46.8% of surgical specimens (44/94), while VEGF-C overexpression occurred in 51.1% of tumor samples (48/94). COX-2 was strongly correlated with VEGF-C expression (P < 0.01), micro-lymphatic vessels (P = 0.032) and metastatic lymph nodes (P = 0. 035). Patients with COX-2 positive tumors had a significant shorter survival time than those with negative tumors did, including disease-free survival (P = 0.010) and overall survival (P = 0.040). Nimesulide could down-regulate the expressions of VEGF-C mRNA and protein in a does-dependent manner, while PGE2 could up-regulate the expressions. The expression of VEGF-C protein up-regulated by PGE2 treatment was decreased by Trastuzumab.

CONCLUSIONSCOX-2 over-expression can up-regulate the expression of VEGF-C. VEGF-C might promote lymph node metastasis by a lymph-angiogenic pathway, then affect the prognosis of the patients with breast cancer.

Adolescent ; Adult ; Aged ; Breast Neoplasms ; metabolism ; pathology ; Cyclooxygenase 2 ; metabolism ; physiology ; Female ; Follow-Up Studies ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Middle Aged ; Prognosis ; Vascular Endothelial Growth Factor C ; genetics ; metabolism