Objective To analyze the changes in the expression of Toll-like receptors 4 (TLR4) and 7 on the surface of murine bone marrow-derived dendritic cells following dengue virus type 2 (DEN2) infection.Methods DEN2 NGC strain was infected into BALB/c suckling mice through intracranial injection and injected into C6/36 cells to induce the in vivo and in vitro proliferation of DEN2, respectively.RT-PCR was performed to identify DEN2 RNA.Reed-Muench method was used to determine the 50% tissue culture infective dose (TCID50) of DEN2.Dendritic cells (DCs) were prepared by stimulating bone marrow cells isolated form C57BL/6 mice with IL-4 and GM-CSF and then identified by flow cytometry.The prepared murine bone marrow-derived DCs were infected with DEN2 and observed with direct immunofluorescence assay.Dynamic changes in the expression of CD11c, CD86 and I-A/I-E molecules on DCs after DEN2 infection were detected by flow cytometry.Levels of DEN2 RNA and the expression of TLR4 and TLR7 at mRNA level were dynamically detected by real-time quantitative PCR.Results The TCID50 of DEN2 to C6/36 cells was 10-5.8.Murine bone marrow-derived DCs were acquired with a purity of 70% and could be infected with DEN2 in vitro.The percentages of CD86 and I-A/I-E molecules on the surface of DCs infected with 1×105 TCID50 of DEN2 were statistically different from those of the negative control group.Neither of the two groups showed a significant difference in the percentages of membrane molecules over time.However, the percentages of membrane molecules on DCs increased with increasing viral load.Compared with the negative control group, the levels of DEN2 RNA in infection groups were increased with increasing virus load, while the expression of TLR4 and TLR7 on DEN2 infected-DCs at mRNA level was decreased with increasing viral load.Conclusion DEN2 infection promotes the maturation of DCs.Expression of TLR4 and TLR7 on DEN2 infected-DCs at mRNA level decreases with increasing viral load, which suggests that TLR4 and TLR7 are closely related to viral infection and play a certain role in the pathogenesis of DEN infection.

Objective:To study the interaction of the inflammatory cytokines expression between CD4+ T cells and primary human umbilical vein endothelial cells (HUVECs) infected by dengue virus (DENV-2).Methods:PBMC was extracted from white blood cells by density gradient centrifugation,CD4+ T cells were sorted by immunomagnetic beads.The expression of CD3 and CD4 molecules on the surface of cells was detected by flow cytometry to identify the purity of CD4+ T cells.First,HUVECs were pretreated by specific-S1 P1 receptor agonist CYM-5442 for 24 h,second,infected by DENV-2 on the titer of 103 TCID50,then co-culturing with CD4+ T cells,The relative expression of NS1 partial sequence and IL-6,L-8 mRNA of HUVECs,and IL-4,IL-17,TNF-α,IFN-γ of CD4+ T cells detected by Real-time RT-PCR.IL-6 and IL-8 secreted in cultured supematant analyzed by ELISA.Results:The purity of CD4+ T cells was (98.02±0.32) ％.The expression of NS1 gradually increased to 24 h (3.03±0.26,P＜0.001),decreased after reaching the peak.The relative expression of NS1 in the group of co-cultured with CD4+ T cells was lower than other groups.After infection,the expression of IL-6 and IL-8 were up-regulated,and the expression of IL-6 and IL-8 at each time point was significantly increased after co-culturing with CD4+ T (P＜0.01).IL-6 of CYM-5442 pretreatment group,in 24 h (28.91±2.34,P＜0.05),36 h(19.36±0.1,P＜ 0.05) and 72 h(13.84±0.82,P＜0.05) was significantly decreased,the expression of IL-8 also decreased significantly.The mRNA expression of IL-4,IL-17,TNF-α and IFN-γ in CD4+ T cells was significantly increased after co-culturing with HUVECs.After the treatment with CYM-5442 group,the expression was decreased.Conclusion:DENV-2 could infect the primary HUVECs,and the expression of NS1 was inhibited after co-culturing with CD4+ T cells.CD4+ T cells can not only enhance the activation of HUVECs infected by DENV-2,but also can be activated by the infected HUVECs infected with DENV-2.

To study the value of high-fluorescent cell for the screening of malignant hydrothorax or ascites. The clinical diagnosis results and hydrothorax or ascites specimens of 400 outpatients registered from January to December in 2014 were collected, and high-fluorescent cells were counted with Sysmex XE-5000 blood cell analyzer, and then stained for microscopy. Receiver operating characteristic (ROC) curve was used to explore the sensitivity, speci-ficity, positive likelihood ratio and negative likelihood ratio of high-fluorescent cell for screening malignant hydrothorax or ascites. The numbers of high-fluorescent cells of the cancer patients were significantly higher than those of the non-cancer patients, with Z equal to -7.372 and P less than 0.05. The values of the area under curve (AUC), sensitivity, specificity, positive likelihood ratio and negative likelihood ratio were 0.801, 70.67%, 84.31%, 4.50 and 0.35 respective-ly. Detection of high-fluorescent may be used to screen malignant hydrothorax or ascites, and further exami-nation and follow-up have to be performed in case the number of high-fluorescent cells in the hydrothorax or ascites is not less than 68.

Vitamine E in peanut oil was determined by means of f Juorometric method (Ex = 285nm, Em=324nm) and TLC scanning method (? = 292nm). Both methods include saponification of the oil, isolation o'f the unsaponifiable matters and the respective final measurements. The result of the former method indicates the total vitamine E in terms of ?-vitamine E and the latter method may indicate their ?,?,? and 6 analogues separately. These two methods are simple, time-saving and reliable. The recovery of VE and the standard deviation of replication of fluorometric method and TLC scanning method are 99.3%, ? 0.587mg/100g sample and 103.04%, ?0.959mg/100g sample respectively. Both methods are also applicable to other foods such as fortified cakes, confectionaries, infant foods etc., but the oil in these samples must be extracted before saponification.

Twenty-three patients with pain from osteoporotic vertebral fractures,aged 65-90 yr,weighing 51-78 kg,received an image intensifier-assisted nerve-root block with a 6-8 ml mixture of 0.5 ％ lidocaine,mecobalamine 0.5 mg and betamethasone sodium phosphate injection 5.26 mg in a prone or lateral position.The VAS scores before operation,at 0,1 week,1 and 3 months after operation were 8.6 ± 0.9,1.5 ± 0.7,2.8 ± 0.9,1.6 ± 0.5 and 2.5 ± 0.7,respectively.VAS scores were significantly lower at each time pint after operation than before operation (P ＜ 0.05).According to modified MacNab standard,the effectiveness of treatment was rated as excellent/good in 87％ of the patients.No complication such as bleeding,hematoma,infection,pneumothorax,hemopneumothorax,headache was found during or after operation.Selective nerve-root block is effective in the treatment of pain resulting from osteoporotic vertebral fractures in patients.

Clinical data of 53 patients with lower caliceal stone during August 2005 and March2007 were analyzed retrospectively.The stones were 11-35 mm in diameter.Under the guidance of X-ray.single renal tract parallel to the lower caliceal for percutaneous nephrolithotomy was established.The procedures were successful in all the patients.Fifty patients were stone free after first minimally invasive pereutaneous nephrolithotomy(MPCNL),2 were stone free following second MPCNL,1 saw residual small stones clear off spontaneously during the follow-up period.Operative time was 65-162 minutes.and blood loss was 10-200 ml.No severe complications or death occurred.MPCNL may be related with minimal invasion and fewer complications,thus provides an effective and safe way of lower caliceal stone treatment.

Objective To investigate the role of norepinephrine in the down-regulated visceral sensitivity of rats deprived of rapid eye movement(REM)sleep.Methods Twenty-four male Sprague- Dawley rats were randomly divided into 3 groups:cage-yoked rats as control(YC),rats with REM sleep deprivation(SD)and rats with yohimbine administered intraperitoneally after REM sleep deprivation(YSD).Flower pot technique was employed to make sleep deprivation model.YSD group was given vohimbine intraperitoneally at the 48th hour after REM sleep deprivation.After both SD and YSD groups had completed these processes,rats of all the three groups were given colorectal distension(CRD)and electromyogram (EMG)was recorded at the same time.The number of discharges of EMG and the threshold of Dain perception of the rats were observed to evaluate the visceral sensitivity.The thalamus,rectum and distal colon were taken after CRD;MAO-mRNA and TH-mRNA in three tissues were detected with RT-PCR.Resuits On 48th hour,the number of discharges of EMG in 10 seconds responding to CRD in group SD was significantly less than that in group YC and the threshold of pain perception in group SD was higher than that in group YC(P＜0.05).The number of discharge of EMG in group YSD was significantly more than that in group SD(P＜0.05).The expression of MAO-mRNA in group SD was lower than that in group YC(P＜0.05)and the expression of TH-mRNA in group SD was higher than that in group YC(P＜ 0.05).Conclusions The visceral sensitivity in rats is down-regulated by REM sleep deprivation,which can increase synthesis of norepinephrine.Norepinephrine can modulate visceral sensitivity.

Objective To investigate the effects of supreme laryngeal mask airway (SLMA) and endotracheal intubation on the elderly hypertensive patients treated with knee arthroplasty. Methods Forty cases of elderly hypertensive patients ASA Ⅰ-Ⅲ treated with knee arthroplasty in our hospital were selected and randomly divided into laryngeal mask airway group (group LMA)and endotracheal intubation (group TT),20 cases for each group.The same protocol for induction and maintenance of general anesthesia was used.After the patients entering,the changes of SpO2 and ECG were performed continuous noninvasive monitoring and SBP,DBP and HR were performed con-tinuous invasive monitoring.SBP,DBP and HR of two groups were recorded at different time points:before anesthesia induction (T0 ,based value),at intubation immediate (T1 ),5 mins after intubation (T2 )and 1 5 mins after intubation (T3 ).At the same time,the content of cortisol (Cor),atrial natri-uretic peptide (ANP)and the concentration of epinephrine (E)and norepinephrine (NE)were meas-ured at the corresponding time points above.Results Compared with T0 ,SBP and DBP at T1-T3 in group LMA were decreased(P <0.05 or P <0.01);SBP and DBP at T1 in group TT were increased while decreased at T2 ,T3 ,HR at T1 were increased(P < 0.05 or P < 0.01 ).Compared with group LMA,SBP and DBP at T1-T3 and HR at T1 ,T2 in group TT were increased (P <0.05 or P <0.01). Compared with T0 and group LMA,the content of E,NE and Cor at T1-T3 increased(P <0.05 or P <0.01).The level of ANP in both groups at T1-T3 were higher than those at T0 ,and group TT were higher than group LMA(P <0.01).Conclusion Compared to endotracheal intubation,SLAM can ef-fectively reduce the stress reaction of elderly hypertensive patients treated with general anesthesia in knee arthroplasty.

Objective To study the depressant effect of atorvastatin on arteriosclerosis of aortic allograft in rats.Methods The models of abdominal aorta transplantation were established in rats with the use of(micro-surgery).The recipients were divided into three groups:allograft control group,allograft experimental group and isograft control group.After 60 days of transplant,vascular intimal thickness(VIT) in all of the groups was observed by histological examination.The expression of PCNA and ?-SMA was determined by(immunohistochemistry).Results The degree of VIT in rats of the allograft experimental group was lower than that in the allograft control group;the VIT area ratio in the allograft control group,allograft experimental group and isograft control group was(12.40?2.65)%,(5.20?6.35)%,and(1.2?1.10)%,respectively,A statistical difference between these groups was observed(P

Objective To evaluate the surgical method and curative effect of minimally invasive percutaneous nephrolithotomy(MPCNL) in treatment of impacted proximal ureteric calculi.Methods 45 cases of proximal ureteric calculi treated with MPCNL were studied retrospectively.Percutaneous nephrostomy was established under the guide of X-ray,B-ultrasound and naked eye,and the nephropathy was extended to F16 through invasive percutaneous.Stones were taken out by ureteropyeloscopic pneumatic lithotripsy with help of hydraulic perfusion pump.Results The surgical time was 15～110 min with the average of 42 min after percutaneous nephrostomy.The average blood loss volume was 30 mL;no severe complications were occurred in the intra-or post-operative time.The rate of complete clearance of stones was 100% in first attempt.The mean hospital stay was 9 d,and the mean postoperative hospital stay was 3.5 d.Conclusion MPCNL used to treat the impacted proximal urethral calculi have satisfied effects and a lot of merits.