The specific fragment of Pneumococcal surface protein A(PspA)and Pneumococcal Surface Adhesin A(PsaA)gene was amplified by PCR from Streptococcus pneumonia 5 and Streptococcus pneumonia 19.The amplified fragnent of PspA and PsaA gene was ligated into pET-27b(+)vector and transformed into BL 21 E.coli for expression and obtain the expressive production of PspA and PsaA.Induced by IPTG,the expression level was as high as 75 % of the total disolube protein.The result showed that the recombinant plasmid could express a specific 75 kDa and 37 kDa fusion protein in E.coli BL 21,which showed the good immunogenicity and a broadly cross reactivity with the other serotypes.

A new flavonoid glycoside, (-)-2S-8-methyl-5,7,4'-trihydroxyflavanone-7-O-β-D-glucopyranoside (1), along with five known ones, quercetin-3-O-(2"-galloyl)-α-L-arabinoside (2), kaempferol-3-O-α-L-arabinoside (3), guaijaverin (4), trifolin (5) and hyperin (6), was isolated from the leaves of Eucalyptus robusta. Their structures with absolute configurations were elucidated by NMR, HR-ESI-MS, CD spectra data and physicochemical methods. In addition, 2-6 were isolated from E. robusta for the first time.
Eucalyptus ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Plant Leaves ; chemistry

Due to the complexity of proteome samples, comprehensive analysis to characterize all proteins was still not possible with present methodologies. It has been shown that replicate runs could increase the number of identified) proteins. However, the redundancy of protein identifications was high. High-abundant peptides tended to be analyzed repeatedly in different runs. To reduce the redundancy and improve the efficiency of identification), we studied the MS/MS acquisition method of linear ion trap Fourier transform ion cyclotron resonance)-mass spectrometry(LTQ-FT) and an acquisition strategy based on exclusion of precursor ions was developed). It proved that the strategy could extremely reduce the redundancy of MS/MS acquisition and improve) the efficiency of protein identifications.

Antigens, CD20 ; metabolism ; CD5 Antigens ; metabolism ; Colonic Neoplasms ; complications ; metabolism ; pathology ; surgery ; Cyclin D1 ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Ileal Diseases ; complications ; pathology ; surgery ; Ileocecal Valve ; Intestinal Neoplasms ; complications ; metabolism ; pathology ; surgery ; Intestinal Polyps ; complications ; metabolism ; pathology ; surgery ; Intussusception ; complications ; pathology ; surgery ; Leukemia, Lymphocytic, Chronic, B-Cell ; metabolism ; pathology ; Lymphoma, Mantle-Cell ; complications ; metabolism ; pathology ; surgery ; Middle Aged

Objective To identify the genotypes of ESBLs-producing Klebsiella pneumoniae isolates from the First Affiliated Hospital,Shantou University Medical College.Methods The MICs of 10 antibiotics were determined by agar-dilution against the clinical isolates of ESBLs-producing K.pneumoniae.PCR were performed with specific primers for blaTEM,blaSHV, blaCTX-M and blaOXA respectively.PCR products were cloned and sequenced.Results The results of PCR showed that a- mong the 83 strains of ESBLs-producing K.pneumoniae,75 were positive for blaTEM,41 positive for blaSHV,25 poitive for blaCTX-M,9 positive for hlaOXA.Three genotypes were found in 13 strains(15.7%),2 genotypes in 59 strains (71.1%) and single genotype in only 11 strains(13.2%).The genes of CTX-M-3,TEM-1 and SHV were found co-existent in 9 strains. The strains carrying 2 or 3 ESBL genes were more resistant to antibiotics than those carrying only 1 ESBL gene.Conclusions The genotypes of ESBLs-producing Klebsiella pneumoniae in this hospital are blaTEM,blaSHV,blaCTX-M and blaOXA. Most strains carry 2 or 3 ESBL genes.

Objective To evaluate the storage performance of storage bags for apheresis platelets produced by Shandong Weigao Group Medical Polymer Co .,Ltd ( experimental bags ) with Trima set platelet storage bags produced by the U .S. Gambro BCT as the control .Method One unit of apheresis platelets was divided into two equal parts , added to control blood bags and experimental blood bags respectively .All samples were stored at ( 22 ±2 )℃ with consecutive oscillation . The platelets′count, mean volume, aggregate activity (ADP, THR), pH, glucose, lactate concentration, lactate dehydro-genase concentration , hypotonic shock reaction , expression of CD62P and phosphatidyl serine on surface of cell membrane were detected at 0,3,5 and 7 d respectively.Results There was no significant difference in platelet quality after five days of storage between the experimental group and the control group (t-test, P>0.05).Conclusion Two types of platelet stor-age blood bags have similar storage performance for apheresis platelets .

Objective To investigate the changes in mitoehondrial membrane potential of the rat neurons after acute brain contusion and laceration injuries. Methods Seventy Sprague-Dawley (SD)rats were randomized into the sham injury group(n=10)and brain injury group.The brain injury group was divided into 6 subgroups(n=10)for observation at 1,6,and 24 h and 2,3,and 7 days after acutebrain contusion and laceration injury,which was induced by impact of the brain with a flee-falling object.In the brain slices labeled with fluorescent JC-l staining,the changes in the mitoehondrial membrane potential of the neurons were observed under confocal laser scanning microscopy at the indicated time points.The neuronal apoptosis Was detected using Hoehesd3342 staining and TUNEL assay,and the number of apoptotic neurons was determined under optical microscope.The ultrastructural alterations of the neurons were observed with transmission electron microscopy. Results Compared with that of the sham injury group,the mitoehondrial membrane potential of the neurons decreased significantly 1 h alterthe brain injury(P<0.01),reachingthe lowestlevel at 24 h,and maintained the low level aRerwards.Apoptotie neurons were identified in the brain slices with Hochest33342 staining.One hour after the brain injury,TUNEL assay revealed a small number of positive neurons,which increased significantly at 6 h(P<0.01)and reached the highest level at 48 h.The alteration pattern of the number of the positive neurons showed a 24-h delay in compariSOIl witll that of the mitochondrial membrane potential aRer the brain trauma.Under electronmicroscope.the mitocbondria of the neurons exhibited only slight swelling at l h. and obvious morphological changes of the mitochondria occurred 6 h after the brain trauma,accompanied by chromatin fragmentation,presence of marginal nuclei and broadened nuclear membrane;nuclear condensation and chromatolysis were observed in the neurons 24 h after the brain trauma.Conclusion In rats with acute brain contusion and laceration injuries,the mitochondrial membrane potential of the neurons decreases early(<1h)after the brain injury,which induces apoptosis of the neurons.

New case is a basic concept in epidemiology and often serves as the numerator for the indexes of the frequency of disease in a population． However，the exact definition of new case is still un- clear． Based on the natural history of disease combined with the concept of event-state，this article rede- fines case-related concepts and compares them with traditional concepts． The study found that the so- called new cases in traditional epidemiological studies are actually the cases firstly discovered，including newly discovered cases and identifiable newly incident cases ( disease initiation) ． In the real world，new cases are often difficult to measure，and together with the continuous process of disease development and iceberg phenomenon，those may affect the accuracy of disease frequency measurement and causal infer- ence． A correct understanding of the meaning of case-related concepts，the meanings they reflect，and the differences between them help to rationally interpret the results of epidemiological research．

Objective To report the phototoxicity effects of a novel photosensitizer ZnPcS4-BSA on photodynamic therapy (PDT) towards human U251 glioma cells in vitro. Methods The cellular uptake of ZnPcS4-BSA by U251 glioma cells was quantified by UV-spectra to determine the optimal incubation time. Human U251 glioma cells were incubated with ZnPcS4-BSA of various concentrations and received laser irradiation of different energy densities. Cell survival rates were measured by CCK-8 assay.Flow cytometer was used to detect apoptosis.Gene expressions of vascular endothelial growth factor (VEGF) were detected by Real-Time PCR in the U251 cells after PDT and β-actin was used as an internal standard. The normal U251 cells severed as controls. Results The uptake of ZnPcS4-BSA by U251 glioma cells reached the maximum after incubation for 4 hours.ZnPcS4-BSA of different concentrations without laser irradiation had no significant effects on cell survival rates (P＞0.05).Without ZnPcS4-BSA incubation,compared with 0,25,50,100,200 J/cm2 groups, the cell survival rate of the 400 J/cm2 group was significantly lower (P＜0.05), whereas no significant difference was found between any other two groups. When the U251 glioma cells incubated with 30 μ mol/L ZnPcS4-BSA for 4 hours underwent laser irradiations of 25,50,100,200 J/cm2,the cellular survival rates significantly decreased with the increased energy densities (P＜0.05). When the U251 glioma cells incubated with ZnPcS4-BSA of 20,40,60,80,100 μ mol/L for 4 hours underwent laser irradiation of 200 J/cm2, the cellular inhibition rates significantly increased with the increased concentrations (P ＜0.05). Compared with controls, the cellular apoptosis and VEGF expression significantly increased in the U251 glioma cells incubated with ZnPcS4-BSA of 20 μmol/L after laser irradiation of 100 J/cm2 (P＜0.05). Conclusion The novel ZnPcS4-BSA is a good photosensitizer for PDT towards U251 glioma cells,because the ZnPcS4-BSA-mediated PDT can induce effective apoptosis of the targeted cells.

OBJECTIVETo study long-term behavioral and ultrastructural alterations in a hypoxic-ischemic brain damage (HIBD) model of neonatal rats.

METHODSSixty seven-day-old Sprague-Dawley rats were randomly subjected to unilateral carotid artery ligation followed by hypoxic exposure (HIBD group) or sham operation (n=30 each). A battery of behavioral tests, including Morris water maze test and sensorimotor tests, were performed at a postnatal age of 5 weeks. Nissl staining was used for counting neurons. Transmission electron microscopy was used for observing synapse structures and measuring the thickness of the postsynaptic density area and the length of the postsynaptic active area. The correlations of histological changes with the results of behavioral tests were evaluated.

RESULTSThe HIBD group showed a significantly longer escape latency (P<0.05) and a lower frequency of original platform crossing (P<0.05) in the Morris water maze test compared with the sham operation group. The sensorimotor function test showed that the sensorimotor function in the HIBD group was worse than in the sham operation group. Nissl staining showed that the number of neurons in the HIBD group was significantly reduced (P<0.01) compared with the sham operation group. Transmission electron microscopy showed that synapses were significantly reduced in number, and that the thickness of the postsynaptic density area and the length of the postsynaptic active area were reduced in the HIBD group. The thickness of the postsynaptic density area was negatively correlated with escape latency in the Morris water maze test (r=-0.861, P<0.01), and also negatively correlated with the total score of sensorimotor function tests (r=-0.758, P<0.05) in the HIBD group.

CONCLUSIONSHypoxia ischemia can lead to neuron loss and ultrastructure damage, resulting in long-term deficit of behavioral functions in neonatal rats.

Animals ; Animals, Newborn ; Brain ; pathology ; ultrastructure ; Female ; Hypoxia-Ischemia, Brain ; pathology ; psychology ; Male ; Maze Learning ; Microscopy, Electron, Transmission ; Rats ; Rats, Sprague-Dawley ; Reaction Time