Objective To evaluate the effect of pancreatic elastase on the expression of TNF ? and IL 1? in Kupffer cells induced by lipopolysacchride. Method Cultivating Kupffer cells were divided into 3 groups. In group A, physiological saline was added into the culture medium as control (control group). Lipopolysacchride (LPS) was added instead of saline in group B (LPS group). In group C both lipopolysacchride and pancreatic elastase were added to the cultere medium (LPS+elastase group). The expressions of TNF ?, IL 1? and TLR4 mRNA in Kupffer cells were determined by RT PCR, and concentrations of TNF ? and IL 1? in the culture media by ELISA. Results The results of both RT PCR and ELISA indicated that the expressions of TNF ? and IL 1? in group C was significantly higher than that in group B. Conclusion It was concluded that pancreatic proteases such as elastase could enhance the expressions of TNF ? and IL 1? of hepatic Kupffer cells induced by lipopolysacchride, and the results the might offer the explanation why inflammatory reaction could be amplified in the course of acute pancreatitis

To reconstruct 3 D images of microstructure and ultrastructure of regenerated sciatic nerve of rats with serial cross sections. The 6 mm defects of the sciatic nerve of 90 Wistar rats were bridge connected with silicone tube and were sampled at 3, 7, 15, 30, 60, 90 days respectively after injury. The images of optical and electronic microscope were taken by micro photograph system and scanner, then they were input into the computer, by which the image registration and segmentation were completed. The direct volume rendering model was used to realize the 3 D reconstruction and display. The results showed the morphological changes in degenerated and regenerated sciatic nerve fibres and their appendicular structures could be observed with the 3 D images. It is suggested that the reconstructing results could reveal the differences between the regenerated and normal sciatic nerve fibres of rats intuitively and visually, so it could be used as a new method in the study of peripheral nerve injury.

Objective To investigate expression of Fos protein in rat brains following restraint stress. Methods The experimental rats were restrained in a small plastic tub for,1,3 and 6 hours,and were sacrificed at 30 min after removing restraint.Immunohistochemical ABC method was used to observe distribution of Fos protein-like immunoreactive(-LI)products in rats brain.Results Fos-LI neurons appeared in (1)Frontal brain:the cingulum,cortex(especially in third and fifth layer),lateral septal nucleus,central amygdaloid nucleus.(2)Diencephalon:the thalamic paraventricular nucleus,lateral geniculate body and medial genicular body,hypothalamic paraventricular nucleus,supraoptic nucleus,periventricular area of third ventricle,arcuate nucleus.(3)Brain stem:the superficial layer of superior colliculus,periaqueductal gray,cortical area of inferior colliculus,lateral parabrachial nucleus,locus coeruleus,A5 area,cochlear nuclei,medullary viceral zone in medulla oblongata.The expression of Fos-LI neurons peaked in rats restrained for 1h,at 3h,then began to decrease,at 6h,significantly decreased. Conclusion Fos-LI neurons appeared in many areas of brain induced with restraint stress.The number of Fos-LI neurons decreased following restraint time.

OBJECTIVETo explore the effect of Buyang Huanwu Decoction (BYHWD) and its disassembled recipes on rats' neurogenesis after focal cerebral ischemia and to investigate its underlying molecular mechanisms.

METHODSFocal cerebral ischemia model was induced by occlusion of the right middle cerebral artery for 90 min using the intraluminal filament model. Rats were divided into the sham-operation group, the model group, the BYHWD group, the qi supplementing group, and the blood activating group. Medication was performed by gastrogavage 24 h after ischemia for 14 successive days. 5-bromodeoxyuridine (BrdU) (at 50 mg/kg) was intraperitoneally injected, once per day for 14 successive days. The neurological function was assessed using modified neurological severity score (mNSS) and the corner test at day 1, 7, and 14 after ischemia. BrdU/Nestin, BrdU/NeuN, and BrdU/GFAP positive cells were examined by double immunofluorescence at day 14 after ischemia. The protein expression of brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF) were detected by Western blot at day 14 after ischemia.

RESULTSCompared with the model group, the score of mNSS and the frequency of turning right significantly decreased in the BYHWD group and the qi supplementing group (P < 0.01), the number of BrdU/Nestin in the subventricular zone of the lateral ventricle, and BrdU/ NeuN and BrdU/GFAP positive cells in the peripheral ischemic cortex increased (P < 0.05, P < 0.01), protein expression of BDNF and VEGF increased (P < 0.05, P < 0.01). In the qi supplementing group, there was no statistical difference in BrdU/GFAP. But there was no statistical difference in each index of the blood activating group (P > 0.05). Compared with BYHWD group, the number of BrdU/Nestin, BrdU/ NeuN, and BrdU/GFAP positive cells significantly decreased (P < 0.01), and the protein expression of BDNF and VEGF were significantly reduced in the qi supplementing group and the blood activating group (P < 0.01).

CONCLUSIONSBYHWD could significantly improve neurogenesis and neurological function recovery after focal cerebral ischemia in rats. Its mechanisms might be related to up-regulating protein expression of BDNF and VEGF. Drugs for qi supplementing and drugs for blood activating had synergistic effects.

Animals ; Brain Ischemia ; drug therapy ; metabolism ; Brain-Derived Neurotrophic Factor ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Male ; Neurogenesis ; drug effects ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; metabolism

AIM: To investigate the effects of recombinant human interleukin-13 (rhIL-13) on the expression of proto-oncogene c-mpl in Dami cells, a human megakaryobiastic leukemia cell line. METHODS: The expression of c-mpl mRNA in Dami cells was investigated with RT-PCR. The expression of membrane-bound protein c-mpl on Dami cells was investigated with ligand combination experiment. RESULTS: In RT-PCR experiment, we found the quantitis of expression of c-mpl mRNA in 25 ?g/L rhIL-13 group increased by 24.8%. In ligand combination experiment, we found quantitis of expression of membrane-bound protein c-mpl in 100 ?g/L rhIL-13 group increased by 28.5% ( P

AIM: Gene transduction with a recombined murine stem cell retroviral vector has been investigated to find an effective way of gene transduction and to offer theory and experimental basis for the recombined murine stem cell retroviral vector used for gene transduction. METHODS: 1. Construction of retrovirus vector: EC1-4 gene (repeats 1-4 of cadherin-5 extracellular domain) and mutant (Ser 222A) MEK1 gene were cloned into retrovirus vector pMSCV after cut by Bgl Ⅱ and EcoR 1 restriction endonuclease. 2. Obtaining CD41 + cells and cell culture: Cells expressing CD34 + from cord blood were isolated. The inducement of cells expressing CD41 from CD34 + cells was performed by using TPO and cells CD41 + were selected by FACS. NIH 3T3 cells were cultured in high sugar DMEM medium and U937 in RPMI 1640 medium. UT7 cells which is cytokine-dependent cell line were grown in Iscove's modified Dulbeco's medium supplemented by GM-CSF. 3. Determination of viral titers: Retroviral vectors were transferred to packaging cell line 293. Retroviral containing supenatant was collected after transfection. The viral titers was tested on infection of NIH 3T3 cells by FACS analysis. 4. Western blot: Transduced CD41 +,UT7,U937 and MDA-MB-435 cells were analyzed by western blot to examine expression of transduced genes. RESULTS: A packaging cell line 293 produces high-titer MEK1 pMSCV retroviruses (3.1?10 7) and EC1-4 pMSCV retroviruses (1.0?10 8). With 8-folds dilution retroviruses,60.73% GFP positive cells have been obtained in MEK1 pMSCV transduced UT7 cells,72.56% in U937 cells and 30.57% in CD41 + cells,respectively. GFP positive cells have reached up 97.54 % in EC1-4 pMSCV transducted MDA-MB-435 cells. Phosphorylated MEK1 has been decreased in experiment group when TPO has stimulated CD41 + and UT7 cells or serum has stimulated U973 cells. This indicates that is a dominant negative effect of mutation MEK gene. EC1-4 gene transduced MDA-MB-435 cells have expressed EC1-4. CONCLUSION: The recombined murine stem cell retrovirus can effectively mediate gene transduction of CD41 +,UT7,U937 and MDA-MB-435 cells,and transduced genes can be stably expressed.

INTRODUCTIONCeftaroline is a fifth-generation cephalosporin with activity against methicillin-resistant Staphylococcus aureus (MRSA) that was recently launched in Singapore. It received approval from the United States (US) Food Drug Administration (FDA) and European Commission for the treatment of adult patients with community-acquired pneumonia (CAP) and complicated skin and soft tissue infections (cSSTI). This study aimed to review current published data and determine its clinical role, particularly in the local setting.

MATERIALS AND METHODSA literature review on published articles in English on ceftaroline, focusing in particular on clinical trials and other clinical reports. Susceptibility testing was also performed on a limited sample of local MRSA and Streptococcus pneumoniae isolates.

RESULTSCeftaroline has an extensive spectrum of activity, including coverage of MRSA and multidrug-resistant S. pneumoniae. However, it has limited activity against non-fermenting Gram-negative bacteria and is susceptible to hydrolysis by extended spectrum beta-lactamases. It is only available for intravenous delivery, with a reconstituted stability of just 6 hours, rendering it unavailable for use for outpatient antibiotic therapy. Clinical trials demonstrate non-inferiority compared to first-line comparators in the treatment of CAP and cSSTI. Published case reports/series suggest a potential greater role in the treatment of MRSA bacteremia and endocarditis. No resistance was found among local archived MRSA and S. pneumoniae isolates.

CONCLUSIONWe believe ceftaroline will occupy primarily niche roles for culture-directed treatment of various infections--in particular those caused by MRSA--until further clinical trial data become available. A variety of factors render it less useful or appealing for empirical treatment of CAP or healthcare-associated infections.

Cephalosporins ; pharmacology ; therapeutic use ; Humans ; Methicillin-Resistant Staphylococcus aureus ; drug effects ; Singapore ; Staphylococcal Infections ; drug therapy

Objective To investigate the source of the blood flow through ventricular septum in normal subject caused by slice-thickness artifact in echocardiography. Methods Echocardiography was performed in 50 normal subjects without ventricular septum defect by two models of echocardiography unit equipped with two models of transducer, observing the conditions and sections in which the blood flow through ventricular septum could be detected. Results The blood flow through ventricular septum was detected in 8 normal subjects using the certain model of echocardiography unit,especially in parasternal four chambers section and parasternal irregular sections, while the blood flow through ventricular septum wasn't detected in the other 42 subjects by any echocardiography unit. The blood flow through ventricular septum was caused by coronary vessel in atrioventrieular groove proved by combining dynamic observation with anatomy analysis. Conclusions The blood flow through ventricular septum in normal subjects, a kind of slice-thickness artifact in echocardiography,is caused by coronary vessel in atrioventricular groove mapped into intact ventricular septum.

CD38 is a multifunctional enzyme expressed in a variety of mammalian tissues, its catalytic activity was involved in a wide range of physiological processes. Based on the reported inhibitor of human CD38 NADase, 33 purine derivatives were designed and synthesized. The biological activity assay showed that compounds 20 and 38 exhibited almost the same extent of inhibitory activities on human CD38 NADase as the lead compound H2. The results also revealed that small substituents at C-6 of purine ring gave no obvious effect on inhibitory activity, but phenylpropionyl moiety at N-2 could affect the binding mode of the compound with CD38. This study provides a reliable basis for future rational design of inhibitors for CD38.
ADP-ribosyl Cyclase 1 ; antagonists & inhibitors ; Enzyme Inhibitors ; chemical synthesis ; chemistry ; Humans ; Purines ; chemical synthesis ; chemistry