Objective To study the effect of moxonidine (Mox) on the His bundle electrogram (HBE) of normal rabbits. Methods A total of 24 healthy rabbits were randomly divided into four groups: control group, small dose of Mox (0.1 mg?kg -1), medium dose of Mox (0.3 mg?kg -1) and large dose of Mox (0.9 mg?kg -1). The electrode catheter was inserted from the right carotid artery to record the HBE. The HBE and the synchronism surface ECG were recorded before and after intravenous injection. Results In normal rabbits, the R-R interphase, P-R interphase of the ECG and the H-V interphase of the HBE were prolonged in a dose-dependent manner after intravenous injection of Mox. Mox exerted no significant influence on the A-H interphase. Conclusion ① Mox decreases the heart rate of rabbits in a dose-dependent manner in vivo. ② Mox dose-dependently prolongs the P-R interphase of the surface ECG and the H-V interphase of HBE. This indicates that Mox mainly acts on the intraventricular conducting system.

Objective To detect eight kinds of clinical common pathogenic bacteria by DNA microarray.Methods Eight kinds of common pathogenic bacteria,including Staphylococcus aureus,Pseudomonas aeruginosa,Klebsiella pneumoniae,Escherichia coli,Proteus mirabilis,Enterobacter aerogenes,Pseudomonas fluorescens,Shigella sonnei were collected.Universal primers were designed to amplify 16S rRNA gene fragment from the genomic DNA of the eight bacteria,and probes were designed in the highly variable regions.DNA microarray detection system was established and used for detection of colleted bacteria.A total of 50 samples were collected from the Zhongnan Hospital of Wuhan University,including 6 blood samples,32 sputum samples,9 feces samples and 3 bronchoscope lavage samples.DNA were extracted and detected by the established DNA microarray system.Results The desired fragments were well amplified by the self-designed universal primers.The selected probes had good detection results according to repeated detection.Of the 50 samples detected,pathgenic bacteria were accurately detected in 47 samples.Other three samples were not detected as those bacteria were not included in the chip.By optimizing the detection process,the results could be reported within 8 hours.Observation of probe signal attenuation indicated that even attenuated after 60 days,but the attenuation did not affect the results.Conclusion A microarray system was established for detection of clinical common bacteria accurately and quickly,which provided foundation for its clinical application.

Objective To develop a gene microarray system for detection of clinical common pathogenic fungi.MethodsThere were 8 clinical common fungi chosen as the subjects including Candida albicans,Candida glabrata,Candida tropicalis,Candida parapsilokis,Candida pseudotropicalis,Aspergillus terreus,Aspergillus flavus,Aspergillus oryzae,Aspergillus fumigatus.Universal primers,probes and specific probes for the PCR amplification and microarray preparation were designed in ITS region of the fungi genomic DNA.The PCR products amplified from those fungi's genome DNA were denatured and hybridized with the probes in gene microarray.The rapid detection of fungi was based on the investigation on the fluorescent signal intensity in the chip.The detection results of gene microarray system were verified by true positive and negative clinical samples.Results There were totally 25 positive samples identified by clinical routine microbiological methods.The 10 samples identified as bacteria positive were determined as negative without fluorescent signal by the fungi gene microarray,while the 12 samples identified as fungi positive were determined as positive with certain fungus by the fungi gene microarray.And 3 artificial Candida krusei samples were detected as fungi positive,while they were failure to be identified as certain fungus.There was no fluorescent signal in positions of the 8 fungi specific probes,but there was fluorescent signal in the position of fungi universal probe.It indicated that there were fungi in the samples but it couldn't identify the species of the fungi,because the Candida krusei wasn't included in the detection fungi list of the fungi gene microarray.ConclusionsThe fungi gene microarray established by the study could detect the common fungi in clinic rapidly and accurately.This study lays technology foundation for clinical application of gene chip.

Objective To detect the expression of VEGF in NHL and analyze the relation of the expression levels with malignant aggressiveness,treatment response,and prognosis.Methods The expression of VEGF in lymph nodes taken from 39 NHL patients Wag measured by immunohistochemical-staining method.9 patients with benign lymphadenopathy were acted as control.Results The expression of VEGF in NHL(79.5%)was higher than that in the contrel(44.4%)(P=0.048＜0.05).In NHL,the VEGF level was higher in aggressive lympboma than that in indolent lymphoma(x2=5.284,P=0.044＜0.05).The patients had the higher-level expression as the Ann Arbor stage Wag higher,and the patients who had the higher-level expression of VEGF had higher serum LDH level,lower chemotherapy remission,unfavorable prognosis and lower 3-year survival (P＜0.05).Conclusion The expression of VEGF in NHL was increased.It was correlated with histopathological grade,Ann Arbor stage,chemotherapy response and prognosis.

Objective To investigate the effect of membrane-bound prostaglandin E2 synthase 1 (mPGES-1) inhibitor MK886 on cell cycle of the human acute myeloid leukemia HL-60/A cells.Methods Flow cytometry,Western blot and ELISA were used to measure the difference of cell cycle,expression of cyclin D1, mPGES-1 among HL-60/A cells,MNC and HL-60 cells.The effect of MK886 on cell cycle,cyclin D1,mPGES-1,PGE2,P-Akt and c-myc of HL-60/A cells were observed.Results Compared with MNC and HL-60 cells,the expression of cyclin D1 and mPGES-1 were higher in HL-60/A cells,the percentage of G0-G1 phase was decreased [MNC (62.63±6.58) ％,HL-60 (38.86±2.25) ％,HL-60/A (30.53±2.15) ％]and S phase increased[MNC (12.18±4.43) ％,HL-60 (47.70±1.88）％,HL-60/A (57.56±1.54) ％](all P＜ 0.05).After treated with MK886,cell cycle was arrested in G0-G1 phase.The expression of mPGES-1,cyclin D1,P-Akt and c-myc and synthesis of PGE2 were decreased.Conclusion MK886 can arrest HL-60/A cell cycles in G0-G1 phase,which possibly through down-regulation of mPGES-1/PGE2,reduction cyclin D1,P-Akt and c-myc expression.

Objectiye To investigate the effect of selective COX-2 inhibitor, nimesulide, on inhibiting proliferation of the human acute myeloid leukemia HL-60 cells. Methods HL-60 cells were treated with different concentration of nimesulide. HL-60 cell proliferation was examined by CCK-8 method. Flow cytometry, Western blotting and ELISA were used to measure the effect of nimesulide on apoptosis, cell cycle,COX-2, PGE2, bax, bcl-2 and c-myc. Results Nimesulide inhibited HL-60 cells proliferation in a dose and time dependence manner. Nimesulide induced cell apoptosis and arrested cell cycle in G0-G1 phase. The expression of COX-2 protein declined after treated with nimesulide 48 h, the total apoptosis in 100, 200,400 μmol/L nimesulide-treated group and control group were (24.97 ± 6.36) %, (34.22 ± 5.76) %, (44.59 ±6.69) % and (4.11 ± 1.26) %, there were significant differences (P ＜ 0.05). Nimesulide inhibited the synthesis of PGE2, the expressions of bcl-2 and c-myc protein and upregulated the expression of bax protein simultaneity.Conclusion Nimesulide significantly inhibited the proliferation of HL-60 cells and induced cell apoptosis,which may be associated with the downregulation of COX-2 expression, reduction of PGE2 synthesis, arrest of cell cycle and regulation bcl-2, c-myc and bax protein expression.

Objective Our preliminary study demonstrates that quercetin can inhibit the proliferation of HL-60 cells. This sudy aimed to find some drugs which could have synergistic effects with quercetin on apoptosis of HL-60 cells. Methods HL-60 cells were cultured with bortezomib at different concentrations (1, 2, 4, 8, 16, and 32μmol/L) alone or combined with quercetin at different concentrations for 48 h. HL-60 cells were cultured with lenalidomide at different concentrations (5, 10, 20, 40, 80, 160, and 320 μmol/L) alone or in combination with quercetin at different concentrations for 48 h. The CCK-8 assay was used to determine the effects on proliferation of HL-60 cells. Results Bortezomib significantly inhibited the proliferation of HL-60 cells (P<0.01). IC50 of quercetin was 49.24μmol/L after cells treated by quercetin combined with bortezomib, which was 13.44μmol/L lower than that treated by quercetin alone. Isobolographic analysis revealed the two drugs had synergistic effect. The results of cell viability of HL-60 cells treated by lenalidomide at lower concentrations (5, 10, 20, 40, and 80μmol/L)were not different from those of the control group (P > 0.05). The results of cell viability of HL-60 cells treated by lenalidomide at higher concentrations (160 and 320μmol/L) were lower than those of the control group (P<0.05). IC50 of quercetin after cells treated by quercetin combined with bortezomib was not different from that treated by quercetin alone. Isobolographic analysis revealed the two drugs had no synergistic effect. Conclusions Bortezomib can inhibit the proliferation of HL-60 cells and it has a synergistic effect with quercetin on HL-60 cells. Lenalidomide has a weaker role in inhibition of the proliferation of HL-60 cells, and it has no synergistic effect with quercetin on HL-60 cells.

Objective To explore the correlation of atheroselerosis progression and the expression of platelet derived endothelial nitric oxide synthase (eNOS) in rabbits. Methods A total of 24 male New Zealand white rabbits were used in this study. Six of the animals were fed with normal food (control group). Eighteen rabbits were fed with cholesterol-rich food (1 g/d) for 12 weeks to establish the atherosclerosis model. Among 18 models, 6 rabbits were executed immediately and their aorta and platelet samples were collected for further analysis (model group), 6 rabbits were orally administered with pravastatin (10 rag/d) for additional 12 weeks (treated group), and the remaining 6 rabbits were left untreated until the end of the study (untreated group). The control, treated and untreated animals were then killed, and the aorta and platelet samples were collected for eNOS expression analysis (RT-PCR). Results The aorta samples in model and untreated group exhibited rough intima and a lot of longitudinal fatty streaks, which indicated that atherosclerosis models were established successfully. While in treated group, the degree of atherosclerosis was decreased. The average percent of thickness of fatty streaks or atheroselerotic plaques relative to the whole thickness of vessel walls was 0. 04±0. 02, 0. 82±0. 16, 0. 33±0. 18,0. 77±0. 14 in control, model, treated and untreated group, respectively. The thickness of fatty streaks or atherosclerotic plaques was significantly increased in the model and untreated groups and decreased in treated group compared with the control group (both P<0. 05). The expressions of platelet derived eNOS/mRNA were 1. 02± 0. 28, 0. 41± 0. 27, 1.00 ± 0. 77, 0. 40±0. 29 in control, model, treated and untreated group, respectively. The expression of eNOS/mRNA was markedly decreased in model group and untreated group compared with the control group, but was increased in treated group compared with untreated and model groups (F=3. 544, P = 0. 024). Conclusions There is a negative correlation between eNOS expression and atherosclerosis development, which suggests that the reversal effect of pravastatin on atheroselerosis progression and plaque formation may relate to the expression of platelet derived eNOS.

Background Corneal neovascularization and inflammation occur in herpes simplex keratitis (HSK).Aciclovir (ACV) is an antiviral medication which is primarily used for the treatment of HSV infection.Bevacizumab is an angiogenesis inhibitor which has the ability to slow the growth of corneal neovascularization.However,whether bevacizumab play treating effects on HSK is worth studing.Objective This study attempted to study the effects of bevacizumab on cornea lesion in mouse models of HSK.Methods The solution containing herpes simplex virus type-1 (HSV-1) of Mckrae strain was induced by cultured and infectious Vero cells and prepared by ten-times step dilution with free-serum DMEM,and plaque assay was used to detect the viral titers.HSV-1 of 1 ×l07 plaque-forming unit (PFU) in 0.6 μl was injected into the corneal stroma of 6 to 8-week-old SPF male C57BL/6 mice using a microliter syringe to establish latent HSK mouse models.The models were examined under the slit lamp microscope at day 5,7,11,14 and 17 after modeling as well as day 0,2,4 and 6 after recurrence,and the central cornea touch sensitivity was recorded.The models were divided into ACV-injected group,ACV+bevacizumabinjected group and normal saline-injected group,and 5 μl normal saline with 50 μg ACV,50 μg ACV + 5 μl bevacizumab or 10 μl normal saline was subconjunctivally iujected according to grouping in 4 eyes of each group,respectively.Twelve model eyes were exposed to ultraviolet (UV)-B to induce the recurrent HSK.Corneal wholemounts were prepared at day 9 after modeling for the assessment of corneal neovascularization and nerve fiber distribution by immunofluorescence assay of CD31 and β Ⅲ Tubulin antibodies.The areas of corneal neovascularization and scarring were mcasured with Image J software.The change rate of lesion was calculated and described as a ratio of lesion size at day 8 with day 0 after induction recurrence.Results The modeling success rate was over 80％,and all infected mice showed latent period at day 45 after modeling.Corneal opacification was the most serious at day 7 after modeling and day 2 after recurrence,and the largest corneal neovascular area was seen at day 15 after modeling and at day 2 after recurrence,and the central cornea touch sensitivity was the worst at day 9 after initial infection.The mean corneal lesion area was 3.348 mm2 in the ACV+bevacizumab-injected group,which was smaller than 3.930 mm2 in the ACV-injected group (Z=-2.309,P =0.021).The central corneal sensitivity in the ACV+bevacizumab-injected group was significantly higher than that in the normal saline-injected group (5.50± 0.71 versus 0.50± 1.41,Z =-2.397,P =0.029).The increase rate of corneal lesion area in the ACV +bevacizumabinjected group was evidently lower than that in the normal saline-injected group ([167.10 ± 52.53]％ versus [312.30± 74.18] ％,Z =-1.992,P =0.046).At the 7th day after modeling,the relative expressing levels of thymidine kinase (TK) and infected-cell protein-27 (ICP-27) mRNA in the corneal tissue and trigeminal ganglion were significantly increased at day 7 and reduced at day 45 after modeling,and the factors raised again at day 2 and retreated at day 7 after induction of recurrence.In addition,the expression of LAT mRNA peaked at day 45 after modeling and reduced gradually at day 2 after recurrence until a new increasing peak at day 7 after recurrence (all at P＜0.01).Immunofluorescence showed that compared with the normal saliue-injected group,the corneal new vessels were lessened and corneal never fibers were increased in the ACV-injected group and ACV +bevacizumab-injected group.Conclusions The combination of bevacizumab with ACV can inhibit corneal neovascularization and scarring in HSK mice,and bevacizumab exhibits a synergistic effect with ACV in management of HSK.

Objective To evaluate the diagnostic value of endoscopic ultrasonography(EUS)for common bile duct(CBD)stones before endoscopic retrograde cholangiopancreatography(ERCP). Methods Data of patients with suspected CBD stones admitted to The First People′s Hospital of Hangzhou from July 2012 to July 2013 were reviewed. Diagnostic efficiency and rates of complications were analyzed between patients undergoing EUS(EUS group)and MRCP(MRCP group)before ERCP, and between patients who underwent EUS and ERCP in different sessions(non-one-session group)with those in one session(one-session group). Results A total of 657 patients were included. With ERCP and follow-up results as the gold standard, the sensitivity(97.5% VS 88.4%), accuracy(96.3% VS 88.0%)and negative predictive value(88.9% VS 60.0%)of EUS in the diagnosis of CBD stones were significantly higher than those of MRCP(P<0.05). There were no significant differences between one-session group and non-one-session group in the sensitivity(97.5% VS 97.4%), specificity(91.7% VS 90.0%), positive predictive value(98.3% VS 97.4%), negative predictive value(88.0% VS 90.0%), and accuracy (96.6% VS 95.9%)in diagnosis of CBD stones(P>0.05). There were no significant differences in incidence of postoperative complications of ERCP between EUS and MRCP group[5.4%(13/242)VS 5.1%(21/415),P>0.05],and between one-session group and non-one-session group[5.5%(8/145)VS 5.2%(5/97),P>0.05].Conclusion Preoperative EUS before ERCP could increase diagnostic sensitivity and negative pridictive value of CBD stones without increasing the incidence of complications.