The aim of this study is to explore the anti-HSV activity of P.vulgaris polysaccharides and gels in vitro and in vivo with the provision of scientific evidence for the further research of anti-HSV new drugs.Plaque reduction assay was adopted to determine the IC50 value of P.vulgaris polysaccharides on HSV-1 and HSV-2 in vitro.In addition,HSV-1 (skin) and HSV-2 (vulva) infected guinea-pig models were established for assessing the anti-HSV activity of polysaccharides and gels in vivo,and infected lesion degree,number of papulovesicle,typical lesion score and the DNA copy numbers of HSV-1 and HSV-2 viruses in the lesion tissue were taken as the indexes.It was found that the activities of HSV1 and HSV-2 viruses was inhibited by P.vulgaris polysaccharides in vitro,while HSV induced skin lesions in the guineapigs were ameliorated by P.vulgaris gel,exerting an anti-HSV action.In conclusion,it was demonstrated that P.vulgaris polysaccharides and gels performed an anti-HSV action both in vitro and in vivo with the hidden value of developing antiHSV agents.

Objective To study the effect and the mechanism of astrocytes-derived estrogen on synaptic transmission. Methods Based on the model of pure cultures of neonatal cortical neurons, the experimental groups were designed as follows: pure neuron culture (group P), ACM culture (group A), mixed culture of AST and neuron (group M), E_ 2 culture (group E), ACM+tamoxifen (estrogen receptor antagonist) culture (group A+T ), tamoxifen culture (group T). The difference among these groups of synaptic transmission was recorded by patch clamp and presynaptic vesicle releasing kinetics was marked by a dye named FM4-64. Results The mean amplitude and frequency of miniature postsynaptic currents (mPSCs) of group P, A, M, E, A+T and T were: (20.5?2) pA, (13?4) min -1 ; (29.1?3) pA, (73?16) min -1 ; (31.3?3) pA, (78?20) min -1 ; (30.2?3) pA, (76?18) min -1 ; (24.5?2) pA, (35?10) min -1 ; (22.1?2) pA, (37?10) min -1 respectivily. The treatment of pure neuron culture with ACM increased synaptic transmission of pure cultured cortical neuron. Exogenic estradiol added into pure neurons can stimulate the effect of the ACM. Tamoxifen which was the antagonist of estrogen receptor could decrease most effect of ACM on synaptic transmission. The assaying of vesicle release marked by FM4-64 showed astrocytes-derived estrogenic enhancing synaptic transmission was at least mediated by strengthening presynaptic vesicle releasing. Conclusion The estrogen from astrocytes of neonatal rat cortex may enhance neuronal synaptic transmission mainly through estrogen receptor.

Objective To study the effect of astrocyte on synapse formation and the molecular mechanism. Methods Cortical astrocytes were isolated and purified from neonatal rats.On the 2h,7th day,14th day and 21th day after passage,we counted the number of astrocytes and the culture medium(astrocyte-conditioned medium,ACM)was harvested to measure the concentration of estrogen(E 2)by using ELISA techniques.Based on the model of pure culures of neonatal cortical neurons,the experimental groups were designed as follows:1.pure neuron cultures(group N);2.ACM cultures(group A);3.mixed cultures(group M);4.E 2 cultures(group E 2);5.ACM+Tamoxifen(estrogen receptor antagonist)cultures(group A+T);6.Tamoxifen cultures(group T).Then synaptic puncta in every group was stained and counted through immunofluorescence,and we also compared the differences in puncta number among those six groups(at 9th day in culture,number/per neuron). Results The numbers of astrocytes were:1?10 4/ml, 1.1?10 6/ml, 1.4?10 6/ml, 1.5?10 6/ml; The concentrations of E 2 were:(ng/L):0, 117?22, 266?22,252?27 respectively.No estrogen was detected in the primary culture medium.The concenteration of estrogen increased in correspondence with the culturing days and reached the peak around at the 14th day, then decreased gradually but kept at a certain high level,and the numbers of synaptic puncta of per neuron in group N,A,M,E 2,A+T,T were:14?3;79?5;83?8;80?6;32?3;29?3 respectively.The treatment of pure neuron culture with ACM increased the number of synapses on per neuron by up to 6 fold by comparison with pure neuronal culture.Exogenic estradiol added into pure neurons can mimick the effect of the ACM.Tamoxifen which is antagonist of estrogen receptor could decrease the effect of ACM by 75%.Conclusion The astrocytes of neonatal rat cortex do secrete E 2.Astrocyte-derived estrogen may be the molecule regulating the synaptic formation through estrogen receptors.

Equipment Design ; Humans ; Nebulizers and Vaporizers

The antioxidant activity of lycopene is highest among the current carotenoids.Recently the researches on lycopene are focused on ingredient of functional foods in the world.The fermentation production of lycopene by Streptomyces rimosus was reported first time in China.The determination methods,such as spectrophotometric method and HPLC were constructed.Using a stain Fc of Streptomyces rimosus as a original strain,a high-yield lycopene producing mutant strain Fc' was selected after UV mutation.The lycopene yield of the strain is 2.5 times higher than the original strain Fc.The optimal fermentation conditions were determined by flask experiments,and the lycopene yield of strain Fc' reached 230mg/L in flask under the optimal fermentation condition,meanwhile the Streptomyces rimosus strain could produce purer lycopene without adding any blocking agents in its fermentation process.This results lay a good foundation for lycopene commercial production by fermentation using Streptomyces rimosus.

Objective To measure the estrogen concentration of estrogen (E 2) in the culture medium of rat astrocytes (ASTs). Methods Astrocytes in brain cortex of the 2-day-old neonatal rats were collected and cultured. The number of astrocytes was counted and the concentration of estrogen was measured by ELISA method at 0, 7, 14, and 21 d after culture. Results The cell counts were 1?10 4/ml, 1.1?10 6/ml, 1.4?10 6/ ml, and 1.5?10 6/ml, respectively. The concentrations of E 2 were: 0 pg/ml, (117.03?21.32) pg/ml, (266.91?22.03) pg/ml, and (252.62?27.99) pg/ml, respectively. No estrogen was detected in the primary culture medium. The concentration of estrogen increased in a time-dependent manner and reached the peak at 14 d, and then decreased gradually but remained at a certain level. Conclusion E 2 is secreted by astrocytes in the brain cortex of the 2-day-old neonatal rats.

OBJECTIVETo study the expression of lipoprotein related genes in subchondral bone of early experimental os-teoarthritis, which may play an important role in the pathogenesis of osteoarthritis.

METHODSAnimals are equally divided into two groups: experimental group and control group, both of which contain fifteen rats of similar weight. The right knee joints of experimental group underwent surgery,which involved in both medial collateral ligament(MCL) transaction and medial meniscectomy, while the control group was only carried out with a sham operation. Rats were killed at 1, 2 and 4 weeks postsurgery to obtain the right knee joints. Total RNA of the subchondral bone was extracted,and then hybridized to Agilent Whole Rat Genome Microarray. Differentially expressed genes analysis was used to study the chemokine signaling pathway.

RESULTSApoa5 expression was down-regulated at 1, 2 weeks post-surgery, Apoc2 expression was up-regulated at 1 week after surgery, Apol3 expression was up-regulated at 1 and down-regulated at 4 weeks post-surgery, Lrp1 expression was down-regulated at 1, 2 weeks after surgery. Lrp5 was down-regulated at 2 weeks after surgery. Gpihbp1, Lpl, Tfpi and Vldlr were up-regulated at 1 weeks after surgery. Lrpap1 and RGD1309808 were down-regulated at 4 weeks after surgery.

CONCLUSIONDysregulation of lipoprotein related genes plays an important role in pathogenesis of early experimental osteoarthritis.

Animals ; Disease Models, Animal ; Knee Joint ; metabolism ; Lipoproteins ; genetics ; Male ; Oligonucleotide Array Sequence Analysis ; Osteoarthritis ; genetics ; Rats ; Rats, Sprague-Dawley ; Transcriptome

ObjectiveTo investigate the molecular epidemiology and mechanisms of carbapenem resistance of Morganella morganii.MethodsSeven carbapenem-non-susceptible M.morganii were isolated from Hangzhou Traditional Chinese Medicine Hospital from October 2010 to February 2011.Pulsed-field gel electrophoresis (PFGE) was performed to analysis the molecular epidemiology of isolates.Antibiotic susceptibilities were determined by agar dilution method.Conjugation experiments were carried out in mixed broth cultures.Plasmid DNA was obtained by an alkalinelysis technique and examined by electrophoresis.Specific PCRs and DNA sequencing were preformed to confirm the genotype of β-lactamases.ResultsPFGE indicated that 6 M.morganii isolates from emergency care unit were indistinguishable or closely related and 1 isolate from intensive care unit was distinguishable.Seven M.morganii showed similar antibiotic susceptibility patterns.M.morganii isolates were resistant to imipenem,were susceptible to meropenem,and were susceptible or intermediate resistant to ertapenem,with MICs of 8 μg/ml,1 μg/ml,and 0.25-0.50 μg/ml,respectively.M.morganii isolates were resistant to penicillins,aztreonam,and ciprofloxacin,were resistant or susceptible to cephalosporins,and were susceptible to amikacin.E.coli (EC600) acquired an approximately 60 kb plasmid from M.morganii by conjugation studies and resistant or intermediate resistant to carbapenems and other β-lactams.PCRs and DNA sequence analysis confirmed that all M.morganii isolates and their E. coli transconjugants produced the KPC-2 carbapenemase and carried the qnrS1 gene.ConclusionIt is the first detection of KPC-2 in M.morganii isolates.Production of KPC-2 mainly contributed to the carbapenem resistance in M.morganii.

Objective To investigate the molecular types of carbapenem-non-susceptible Esche-richia coli ( E.coli) isolates and their mechanism of carbapenem resistance .Methods Twenty-two carbap-enem-non-susceptible E.coli strains were isolated from 3 hospitals in Hangzhou from 2007 to 2011.The mini-mum inhibitory concentrations ( MICs) of antimicrobials to those isolates were determined by agar dilution method and E-test.The molecular mechanisms of carbapenem resistance of E.coli isolates were analyzed by conjugation experiment,PCR and DNA sequencing.Pulsed-field gel electrophoresis (PFGE),multilocus se-quence typing ( MLST ) , and phylogenetic typing were performed to analyze the molecular epidemiology of those isolates.Results The MICs of imipenem and meropenem to 22 E.coli isolates were ranged from 1 μg/ml to 16 μg/ml,and the MICs of ertapenem were 2 μg/ml to 64 μg/ml.All E.coli isolates produced the KPC-2 carbapenemase and various β-lactamases , and some of them also produced plasmid-mediated AmpC enzymes.Carbapenem resistance was transferred by conjugation and transformation from 22 E.coli iso-lates to E.coli EC600 strains.The E.coli transconjugants or transformants acquired the blaKPC-2 gene and showed similar antibiotic susceptibility patterns in comparison with donor strains .Only a few isolates were in-distinguishable or closely related as indicated by PFGE .Four sequence types including ST131 (9 isolates), ST648 (5 isolates),ST38 (2 isolates) and ST405 (2 isolates) were identified by MLST.Phylogenetic analy-sis indicated that 9 ST131 isolates belonged to phylogenetic group B 2 and the other isolates belonged to group D (11 isolates),group B1 (1 isolate) and group A (1 isolate),respectively.Conclusion The sequence type of prevalent E.coli isolates producing KPC-2 from Hangzhou was ST131,which is an international epi-demic,multidrug-resistant clone,followed by ST648.