Objective To investigate the relationship between heme oxygenase-1 (HO-1),glutathione S-transferase (GST) and cerebral atherosclerosis.Methods Cerebralvascular status was assessed with color flow Doppler sonography,transcranial Doppler (TCD),magnetic resonance angiography (MRA)or/and digital subtraction angiography (DSA) in patients with cerebral atherosclerosis (mild,moderate,and severity).Serum HO-1 and GST were measured with enzyme-linked immunosorbent assay (ELISA).Results In comparison between case and control groups,there was significant difference in age,hypertension,cerebral infarction,uric acid,and HO-1 (P =0.041,0.008,0.000,0.036,and 0.001).The level of serum HO-1 in the severe atherosclerosis was lower than that in the mild and moderate atherosclerosis (P =0.000 and 0.002).Logistic regression was used to find the association of HO-1 and the degree of cerebral atherosclerosis (P =0.000).Conclusions HO-1 might be related to cerebral atherosclerosis.

Background Retinal hypoxia is one of primary causes of retinal neovascularization,and its mechanism is research hot topic.Studies showed that hypoxia stimulates the activation of many trancripation factors and vascular endothelial cells,which leads to angiogenesis.Besides to the vascular endothelial growth factor,the effect of adenosine on angiogenesis is increasingly concerned.Objective This study was to invastigate the relationship of biological behaviour of human microvascular endothelial cells (HMEC-1) under the hypoxia and adenosine,and to explore the effect of adenosine on angiogenesis under hypoxia.Methods HMEC-1 cell line was cultured in vitro,and the cells were divided into normoxia group and hypoxia group.The cells in the normoxia group were cultured under the 5％ CO2 environment,and those in the hypoxia group were cultured under the 1％ O2,94％ N2 and 5％ CO2 environment.The proliferation ability and percentage of the cells were assayed by cell counting kit-8 (CCK8) and EDU.The migration number and invasive number of the cells were detected by transwell chamber.The expressions of CD39 and CD73 proteins in the cells were tested by Western blot and immunofluorescence technique,and the level of adenosine was measured by high performance liquid chromatography (HPLC).Results The proliferation values (absobancy) were 0.715-±0.067 and 0.821 ±0.056 in the normoxia group and the hypoxia group in 12 hours after culture,and those in 24 hours were 0.946±0.028 and 0.998±0.028,showing significant increase in the hypoxia group compared with the normoxia group (t12h =3.805,t24h =3.222,all at P ＜ 0.01).The precentage of the proliferation in the hypoxia group was evidently higher than that in the normoxia group (t =-6.868,P＜0.01).The number of the cell migration and invasion in 24 hours after culture was 185.3 ± 10.594 and 74.2± 10.741 respectively in the normoxia group,and that in the hypoxia group was 300.7±22.853 and 107.5±7.007,with significant differences between the two groups (t=-12.124,-6.367,both at P＜0.01).The expression levels of CD39 and CD73 proteins in the hypoxia group were significantly higher than those in the normoxia group in bothl2 hours and 24 hours after culture(all at P＜0.05),and the adenosine content in the cells is gradually increased in 2,6,12,24 and 36 hours after culture,with the highest content in 36 hours.The adenosine content was significantly higher in the hypoxia group than in the normoxia group at various time points (t2h =2.469,P =0.017;t6h =5.442,P＜0.001;t12h =3.841,P＜0.001;t24h =4.458,P＜0.001;t36h =2.757,P =0.008;t48h =3.319,P =0.002).Conclusions Compared with the normoxia group,the proliferation,migration and invasion abilities of HMEC-1 are stronger,meanwhile,the expression of CD39 and CD73 as well as the adenosine level in the cells are all increased under the hypoxic condition.It is suggested that the activation of human microvascular endothelial cells might be significantly related to the level of adenosine and its key enzymes.

Objective To compare clinical effect and safety of left-double-lumen tube and bron-chial blocker in vedio-assisted atrial fibrillation ablation.Methods Forty-eight patients,26 males and 22 females,aged 45-65 years,ASA physical status Ⅰ or Ⅱ,who underwent vedio-assisted atrial fi-brillation ablation were randomly divided into double-lumen tube (group A)and bronchial blocker (group B),with 24 patients in each group.The tube type of group A was left-double-lumen tube. Bronchofiberscope was used for location in every patient.The mean artery pressure (MAP)and heart rate (HR)before intubation,intubation positioning time,peak airway pressure (Ppeak)after 5 min of one-lung ventilation,lung collapse,incidences of hoarseness,pharyngalgia and choke were ob-served.Results Intubation positioning time between two groups was not statistically significant. MAP and HR were significantly increased at intubation positioning time in both groups,to be specif-ic,they were significantly in group A than in group B (P <0.05 ).When left lungs blocked,Ppeak and qualities of lung collapse were not statistically different between the two groups.When right lungs blocked,group A was higher than that in group B (P < 0.05 ).Cough,hoarseness and sore throat were more frequently seen in group A than in group B.Conclusion Both double-lumen tube and bron-chial blocker can be used in video-assisted atrial fibrillation ablation with satisfying effects.As for the quality of lung isolation,double-lumen tube was better than bronchial blocker.However,compared with bronchial blocker,double-lumen tube results in more unstable hemodynamics and higher occur-rence of hoarseness,pharyngalgia and choke.

Objective:To investigate the Repetitive transcranial magnetic stimulation combined with biofeedback non motor symptoms on clinical effects and serum levels of IL-6,CRP and TNF-α of Parkinson's disease.Methods:84 cases of Parkinson's disease in our hospital were randomly divided into experimental group and control group,with 42 cases in each group.The control group were treated with EMG and EEG biofeedback,while the experimental group were treated with repetitive transcranial magnetic stimulation on the basis of the control group.Then the serum levels of C reactive protein (CRP) and interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α),the non motor symptoms questionnaire (NMSQuest),Parkinson's disease Sleep Scale (PDSS) and Hamilton Depression Scale (HAMD) score in the two groups were observed and compared before and after the treatment.Results:Compared with before treatment,the serum levels of CRP,IL-6 and TNF-α,the NMSQuest and HAMD in the two groups decreased after the treatment,while the PDSS score increased,and the differences were statistically significant (P＜0.05);Compared with the control group,the serum levels of CRP,IL-6 and TNF-α,NMSQuest and HAMD in the experimental group were lower,while the PDSS score was higher,and the differences were statistically significant (P＜0.05).Conclusion:Repetitive transcranial magnetic stimulation combined with biofeedback can reduce the serum levels of IL-6,CRP and TNF-α in patients with Parkinson's disease,which has better clinical effect on the non motor symptoms.

OBJECTIVE:To establish a method for the contents determination of emodin,rhein,aloe-emodin,chrysophanol, physcion,geniposide,chlorogenic acid in Yinchenhao decoction and its dispensing granule,and to compare the difference among the above-mentioned ingredients. METHODS:HPLC was adopted to determine the contents of emodin, rhein, aloe-emodin, chrysophanol and physcion:the column was Ecosil C18 with mobile phase of acetonitrile-0.5% phosphoric acid(gradient elution)at a flow rate of 1 ml/min,the detection wavelength was 254 nm,column temperature was 35 ℃,and injection volume was 20 μl. When determining the content of geniposide by HPLC,the column was Ecosil C18 with mobile phase of acetonitrile-water(13∶87, V/V)at a flow rate of 1 ml/min,the detection wavelength was 238 nm,column temperature was 35 ℃,and injection volume was 20 μl. When determining the content of chlorogenic acid by HPLC,the column was Ecosil C18 with mobile phase of acetonitrile-0.5% phosphoric acid (10∶90,V/V) at a flow rate of 1 ml/min,the detection wavelength was 327 nm,column temperature was 35 ℃,and injection volume was 20 μl. RESULTS:The linear range was 2-20 μg/ml for emodin(r=0.996 5),5.2-52.0 μg/ml for rhein(r=0.998 5),2.6-26.0 μg/ml for aloe-emodin(r=0.999 9),1.0-10.4 μg/ml for chrysophanol(r=0.999 9),1.0-10.0 μg/ml for physcion(r=0.999 8),20-200 μg/ml for geniposide(r=0.999 9),20-200 μg/ml for chlorogenic acid(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 3%;recoveries were 91.1%-96.9%(RSD=2.0%,n=6)for emodin、93.9%-96.1%(RSD=0.8%,n=6)for rhein、90.9%-93.4%(RSD=1.2%,n=6)for aloe-emodin、88.5%-92.7%(RSD=1.8%,n=6) for chrysophanol、82.1% -87.9%(RSD=2.5% ,n=6)for physcion,100.4% -102.0%(RSD=0.7% ,n=6)for geniposide、101.1%-102.2%(RSD=0.4%,n=6) for chlorogenic acid. CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the simultaneous contents determination of active ingredients in Yinchenhao decoction and its dispensing granule. The contents of active ingredients in Yinchenhao dispensing granule are obviously higher than those in its decoction.

Objective To evaluate the efficacy of continuous lumbar plexus block (CLPB) combined with a bolus dose added at night for postoperative analgesia in patients undergoing hip arthroplasty.Methods Sixty ASA Ⅰ or Ⅱ patients of both sexes,aged 51-75 yr,weighing 47-77 kg,with body height 150-180 cm,scheduled for hip arthroplasty,were randomized to receive either CLPB (group CLPB) or patient-controlled intravenous analgesia (PCIA) (PCIA group) for postoperative analgesia (n =30 each).Spinal anesthesia was performed at L3,4 interspace.Postoperative analgesia was performed at 30 min before the end of surgery.PCIA solution contained morphine 100 mg in 100 ml of normal saline.The PCA pump was set up with a 2 mg bolus dose and a 5 min lockout interval.CLPB solution contained 0.125 ％ ropivacaine hydrochloride 200 ml.CLPB pump was set up to deliver a 4 ml bolus dose with a 30-min lockout interval and background infusion at 8 ml/h after a loading dose of 0.125％ ropivacaine 30 ml.In addition the patients received 0.25％ ropivacaine 30 ml at 8 o' clock every night after surgery in group CLPB.VAS scores at rest and during activity were recorded at 6,12,18,24,30,36,42 and 48h after operation.The side effect such as nausea and vomiting,pruritus and urinary retention were recorded within 48 h after operation.The patient' s satisfaction was assessed.The maximal hip flexion and abduction ranges of motion were recorded at 12,24,36 and 48 h after operation.The times of sleep interruption resulted from pain during nighttime were also recorded.Results Compared with group PCIA,the VAS scores during activity,severity of nausea and vomiting,pruritus and urinary retention,and times of sleep interruption resulted from pain during nighttime were significantly decreased,and the overall satisfaction score and maximal hip flexion and abduction ranges of motion were increased in group CLPB (P ＜ 0.05).Conclusion CLPB combined with a bolus dose added at night can provide better efficacy for postoperative analgesia in patients undergoing hip arthroplasty than PCIA,with fewer complications.

Background Retinal ganglion cell (RGCs) death following ischaemic insult is the major cause of a number of vision-threatening diseases.Recent studies confirmed that micro RNA (miR-30b) can alleviate hypoxy-induced cardiac injury.However,whether miR-30b can protect RGCs against oxygen-glucose deprivation damage is still not ellucidated.Objective The aim of this study was to investigate the protective effect of miR-30b on RGCs damage caused by oxygen-glucose deprivation.Methods The retinas were isolated from the eyeballs of eight SD rats aged postnatal 24 hours and RGCs were primarily cultured.The cells were divided into the recombinant adeno-associated virus (rAVV) control group,rAAV-miR-30b mimic group and AAV-miR-30b inhibitor group.Then the cells were transfected using rAVV-miR plasmid,rAAV-miR-30b mimic plasmid and AAV-miR-30b inhibitor plasmid,respectively for 6 days with the RGCs ∶ AAV as 1 ∶ 10 000.The cells were cultured with low glucose medium in hypoxygen incubator (5％ CO2,17％ N2,3％ O2) or 5％ CO2 incubator respectively for 24 hours.Cell viability was detected by cell counting kit-8 assay.The expression of Tubulin Ⅲ,a neuron specific marker,was detected by immunofluorescence technology to evaluate the survival of RGCs.The apoptosis and necrosis of the cells were assessed by Hoechst/PI double staining.Results The RGCs grew well with round shape and 1 3 processes 7 days after cultured in the normal cells.However,the RGCs were diminished and the cell process disrupted in the oxygen-glucose deprivation group.The relative vability of the cells was 3.310-±0.162 in the rAAV-miR-30b mimic group,which was significantly higher than 0.949±0.141 in the rAAV-miR-30b inhibitor group and 0.900±0.181 in the rAAV-miR control group(t=10.508,10.296,both at P＜0.001).It was positively expressed in survival RGCs,with the red fluorescence.The number of Tubulin Ⅲ+ cells was (13.800± 1.924)/field in the rAAV-miR-30b mimic group,showing a significant increase in comparison with (0.600±0.548)/field in the rAAV-miR-30b inhibitor group and (0.800± 1.304)/field in the rAAV-miR control group (t =15.141,14.912,both at P ＜ 0.001).Significant differences were found in the apoptosis rate and necrosis rate among the rAAV-miR-30b mimic group,rAAV-miR control group and PBS group (F=10.851,P=0.002;F=6.378,P=0.013),and the apoptosis rate and necrosis rate in the rAAV-miR-30b mimic group were considerably lower than those in the rAAV-miR control group and PBS group (all at P＜0.05).Conclusions The oxygen-glucose deprivation models can be established in RGCs by hypooxygic and low-glucose cultivation.rAAV encoding miR-30b mimics transfection can protect RGCs against oxygen-glucose deprivation damage.

Objective To investigate the effects of contrast dose and region of interest (ROI) depth on quantitative analysis of liver by contrast-enhanced ultrasound (CEUS) during clinical application.Methods After bolus injection of contrast agent,the change of quantitative parameters [including echo mean(EM),rise time(RT),peak intensity(PI),mean transit time(MTT),area under the curve(AUC),time from peak to one half(TPH),wash in slope(WIS),time to peak(TTP)] of time-intensity curves were analyzed based on groups from different doses (1.0 ml and 1.6 ml) and different depth (＜30 mm,30-60 mm,and ≥60 mm).Results MTT and TPH were increased with dose increasing from 1.0 ml to 1.6 ml (P＜0.05).With the dose 1.0 ml,TPH,WIS,PI,AUC and MTT showed significant difference when the depth of the ROI changes (P ＜0.05),with the depth increased,TPH,WIS,PI,and AUC all decreases and MTT increases.For all the other parameters,no significant changes were found (P ＞0.05).Conclusions CEUS and its imaging process can directly influence the accuracy of the parameters from the quantitative analysis.Standardization of contrast agent with predefined dose and depth can potentially facilitate future clinical studies in liver CEUS.

Objective To investigate the application of contrast-enhanced ultrasonograhy CEUS in the early assessment of antiangiogenic treatment in metastatic colorectal cancer Methods Among 41 colorectal cancer patients with liver metastases enrolled in this study 25 patients received treatment of antiangiogenic treatment bevacizumab combined with chemotherapy BV group and 16 patients received chemotherapy only non-BV group To assess the treatment responses in the liver lesions between and within these two groups CEUS scans were performed before the treatment T0 or??baseline on the day 2 T1 and on the day 7 T2 after the treatment The percentage differences were evaluated according to the CEUS quantitative parameters such as rise time RT peak intensity PI mean transit time MTT area under the curve AUC time from peak to one half TPH wash in slope WIS and time to peak TTP Results On T1 both the RT and MTT increased within the BV group and decrease within the non-BV group when compared with their own baselines the differences were statistically significant P <0 05 On T2 the MTT increased within the BV group and decreased within the non-BV group when compared with their baselines the differences again were statistically significant P <0 05 All other parameters measured on T1 and T2 did not show a significant difference when compared with baselines Within the BV group the RT decreased and WIS and TPH increased for the responding subjects when compared with the non-responding subjects the differences were statistically significant P < 0 05 Within the non-BV group however no statistically significant differences were found for all parameters between the responding subjects and non-responding subjects P >0 05 Conclusions CEUS can be used to assess the treatment response in liver metastatic patients with colorectal cancer by monitoring the change of the tumor perfusion before and after the treatment.

ABSTRACT:Objective To explore the effects of miRNA-1246 (miR-1246)on cell proliferation,invasion and migration in human cervical squamous cell carcinoma (CSCC)cell line SiHa.Methods SiHa cells were assigned into 3 groups:miR-1246 analog group,miR-1246 antagonist group and control group.Transfection efficiency was determined.The MTT assay,transwell assay and wound healing assay were performed respectively to evaluate the proliferation,invasion and migration abilities of SiHa cells.Western blot was carried out to detect the expression of thrombospondin-2 (THBS2)before and after transfection.A THBS2 3’-UTR-containing dual luciferase plasmid was synthesized and co-transfected with miR-1246 into SiHa cells to observe the luciferase enzyme activity.Results MTT assay,transwell assay and wound healing assay revealed that the abilities of proliferation,migration and invasion were significantly enhanced (P<0.01)in SiHa cells transfected with miR-1246 analog,but suppressed in SiHa cells transfected with miR-1246 antagonist.Western blot showed that SiHa cells transfected with miR-1246 analog had significantly decreased THBS2 expression (gray value = 6 .2 8 ± 1 0 .2 2 , P=0 .0 1 3 ) while those transfected with miR-1246 antagonist had significantly increased THBS2 expression (gray value = 12.90±19.81, P=0.037).After co-transfected with miR-1246 and THBS2 3’-UTR-containing plasmid,SiHa cells exhibited a decreased level of luciferase enzyme expression.Conclusion miR-1246 promoted the proliferation,invasion and migration of CSCC SiHa cell, and it might promote CSCC tumorigenesis and progression by suppressing the expression of its target gene THBS2 .