Objective To observe the process of Vibrio vulnificus inducing dendritic cell strain apoptosis.Methods We established the mixed culture model of mouse dendritic cell ( DC 2.4 strain) and Vibrio vulnificus( Vv1.1758 strain ),analyzed morphological characteristics of cell apoptosis by DAPI fluorescence staining,detect DNA fragmentation level of apoptosis cells by DNA Ladder assay,analyze DC2.4 apoptosis rate by Annexin V FITC/PI staining,determine activities of caspase-3 and caspase-8 by means of spectrophotometric method and detect changes of mitochondrial transmembrane potential ( △ Ψm ) by JC-1 fluorescence labeling.Results After Vv1.1758 strain and DC2.4 cell were mixed and cultured for 4 h,DAPI fluorescence staining showed typical apoptosis characteristics-chromatin condensation and marginalization; DNA agarose gel electrophoresis showed apoptosis band; apoptosis rates at 2,4 and 6 h were respectively (37.8±9.8) ％,(54.3 ± 12.7 ) ％ and ( 68.2± 14.6 ) ％ ; Mitochondrial transmembrane potentials (△Ψm) at 1 h,2 h and 4 h reduced by 7.1％,16.1％ and 46.7％ respectively; caspase-8 activity increased at 1.5 h and reached the peak at 2 h [ (2.48±0.19) U/μg],while caspase-3 activity started to increase at 3h and reached the peak at 4 h [ ( 1.91 ±0.16) U/μg ].Conclusion Vibrio vulnificus could induce dendritic cells by two pathways: reducing mitochondrial transmembrane potential and activating caspase-8 promoter and finally activate effector caspase-3 to promote apoptosis.

Objective To analyze the association of blood uric acid level with the severity of coronary artery stenotic changes, metabolic syndrome (MS), and its components. Methods A total of 343 individuals ( male 223,female 120) who underwent coronary angiography and had complete data on MS and serum uric acid were collected. The severity of coronary artery disease (CAD) was assessed by the coronary stenesis index (CSI). MS was diagnosed according to the Guideline on Prevention and Treatment of Blood Lipid Abnormality in Chinese Adults. Results (1)The mean uric acid level was significantly lower in women than in men [ ( 306.3±76.9 vs 358.9±85.2 ) μmol/L, P＜0.01 ]. The prevalence of MS and its components showed no difference between men and women. (2) The uric acid level in women with 3 components was higher than those with1( P＜0. 01 ) or 2 ( P＜0.05 ) components of metabolic disorders, but not in men. (3) Quartiles of concentration of uric acid were computed. Compared with those in the lowest quartile of uric acid, women in the highest quartile had higher CSI score [ 7.0 (2.5-12.0) vs 2. 0( 0.0-6.0), P= 0. 025 ]. Moreover, the uric acid level was higher in women with multivessel lesions than nonCAD patients [ (327.0±81.9 vs 284.9±78.6) μmol/L, P = 0.033 ]. However, no correlation was found between uric acid level and the severity of coronary artery lesion in men. (4) Logistic regression showed that age (β=0.042, P=0. 007) and dyslipidemia(β=0.836, P=0. 037 ) were the independent risk factors of CAD in men, and hypertension(β=1. 127, P=0.039) and dyslipidemia(β=0.901, P=0.009)in women. Conclusions In women with higher uric acid level, the clustering of metabolic abnormalities was increased, and the coronary artery lesion was more severe. High uric acid level might be a marker of CAD for women.

Objective To explore the clinical value of AFP-L3,GP73 and GGT as biomarkers in diagnosis of hepatocellular carcinoma (HCC).Methods According to the pathological diagnosis,141 patients were divided into two groups,HCC group were 74 cases,benign liver disease group were 67 cases.Use ELISA method tested the serum AFP-L3 and GP73 levels.The GGT level was detected by the automatic biochemical instrument of all the 141 patients.AFP-L3,GP73 and GGT concentration difference was compared between the two groups.ROC curve was used to determine the cut-off level to diagnose HCC.The value of single use AFP-L3,GP73,GGT and joint the three indexes to diagnose HCC were analyzed.Results The average level of AFP-L3 in the patients with HCC was (113.58±63.62) μg/L,it was significantly higher than that in the patients with benign liver diseases [(23.19±34.54) μg/L] (P ＜ 0.001).The area under the ROC curve of AFP-L3 level was 0.802.Taking AFP-L3 level ≥ 38.47 μg/L as diagnostic criteria,the sensitivity of AFP-L3 level in HCC diagnosis was 81.08 ％ and the specificity was 88.06 ％.The average level of GP73 in the patients with HCC was (126.55±49.56) μg/L,it was significantly higher than that in the patients with benign liver diseases [(56.97±26.48) μg/L] (P ＜ 0.001).The area under the ROC curve of GP73 level was 0.811.Taking GP73 level≥69.44 μg/L as diagnostic criteria,the sensitivity of GP73 level in HCC diagnosis was 75.68 ％ and the specificity was 91.04 ％.The average level of GGT in the patients with HCC was (173.20±179.18) U/L,it was significantly higher than that in the patients with benign liver diseases [(90.77±81.53) U/L] (P ＜ 0.001).The area under the ROC curve of GGT level was 0.713.Taking GGT level ≥ 110.77 U/L as diagnostic criteria,the sensitivity of GGT level in HCC diagnosis was 74.32 ％ and the specificity was 77.61％.Joint use AFP-L3,GP73 and GGT to diagnose HCC,the sensitivity was 83.78 ％,specificity was 92.53 ％.Conclusion Combined detection of tumor markers AFP-L3,GP73 and GGT can improve the positive rate of HCC,which has good clinical application value.

Objective:To investigate the expression of wild type estrogen receptor(wER)and the ex-on-5 deleted ER(variant ER,vER)in human hepatocellular carcinoma(HCC)samples,and thereafteranalyze the possibility of HCC treatment by endocrine therapy.Methods:The mRNA expressions of wERand vER were analysed from 28 cases of HCC by RT-PCR.The expression of ER at the protein level wasdetected by immunohistochemistry(IHC).Results:IHC results showed that 39.3% of the HCC speci-mens expressed ER.The mRNA of wER was detected in 89.3%(25/28)of the HCC specimens whilethat of vER was detected in 96.4%(27/28).Twenty four out of 28 HCC cases(85.7%)expressedboth wER and vER.One out of 28 patients(3.5%)expressed only wER whereas 3 patients out of 28(10.7%)expressed vER only.Conclusion:Ninety six percent(27/28)of the HCC patients expressedvER,which suggests that the expression of vER is an important event in the development of HCC.

Aim To study the effect of Baoxinbao film on endothelin(ET) and nitric oxide(NO) secretion in patients with stable angina pectoris(SAP).Methods 76 patients with SAP were randomly divided into two groups, with 40 cases in the baoxinbao group plastered with baoxinbao film and 36 cases in the isosorbide dinitrate group receiving isosorbide dinitrate. The levels of plasma ET and NO before and after treatment were observed. Results The concentrations of plasma ET were increased and plasma NO reduced significantly in the SAP patients respectively, as compared with those in the control group(all P

Objective To explore the role of clinical pharmacists in rational drug use through the pharmacy care of an elderly pneumonia patient with Chlamydia psittaci infection and drug-induced liver injury. Methods The clinical pharmacists participated in the treatment of one patient with Chlamydia psittaci pneumonia and drug-induced liver injury. Based on the results of second-generation gene sequencing, the characteristics of the pathogen were learned by literature search. The clinical pharmacists monitored the patient’s liver and kidney function, provided a new medication treatment plan to Doctors, and performed patient education during the treatment. Results The initial empirical anti-infective treatment with teicoplanin and imipenem-cilastatin was not effective. After the diagnosis of Chlamydia psittaci and Candida albicans infection, the combination of doxycycline with azithromycin and fluconazole was administered. Drug-induced liver injury was found with this treatment. The clinical pharmacist proposed to switch to doxycycline and clarithromycin with co-administration of magnesium isoglycyrrhizinate and polyene phosphatidylcholine to protect the liver. With this new regime, patient's liver function was improved and the infection was under control. Conclusion Individualized pharmaceutical cares provided by clinical pharmacists helped the safe, rational and effective use of medications.

Objective To evaluate collateral flows using vessel encoded arterial spin labeling (VE-ASL) perfusion imaging. MethodsVE-ASL was achieved to assess the presence and function of collateral flow on patients with internal carotid artery (ICA) stenosis. The presence of the anterior and posterior collateral flow was demonstrated by flow patterns of the A1 segment and posterior communicating artery (PCoA).Distal function of collateral flow of stenotic hemisphere was categorized as adequate ( cerebral blood flow ≥10 ml · min-1·100 g-1 ) or deficient (cerebral blood flow < 10 ml · min-1· 100 g-1 ). The results were compared with magnetic resonance angiography (MRA) and intraarterial digital subtraction angiography (DSA) in crosstable by using Kappa values. The VE-ASL before and after ICA stent therapy were compared. ResultsThe Kappa values of the flow patterns of AI segment and PCoA between VE-ASL and MRA were 0. 746 and 0. 700. The Kappa value of the function of collaterals using VE-ASL and DSA was914. VE-ASL showed collateral flow via leptomeningeal anastomoses. VE-ASL changed significantly after ICA steat therapy. ConclusionVE-ASL reveals the presence and distal function of collateral flow, which helps to evaluate the efficacy of ICA steat therapy.

BACKGROUND AND OBJECTIVENitric oxide (NO) is involved in many physiologic and pathologic processes. As an important biologic mediator, NO has been the focus of cancer study for its function in tumorigenesis, tumor progression, and death. This study investigated the effect of NO donor sodium nitroprusside (SNP) on the growth and proliferation of gastric cancer cell line AGS.

METHODSThe growth inhibition of AGS cells was analyzed using MTT assay. The cell cycle was measured using flow cytometry. The changes of mRNA expression of proliferating cell nuclear antigen (PCNA) and caspase-3 were examined using reverse transcriptase polymerase chain reaction (RT-PCR), and the protein expressions of PCNA and caspase-3 were analyzed using Western blot.

RESULTSDose-dependent SNP inhibited cell growth and proliferation. When the AGS cells were treated with SNP at 100, 500, 1000, 1500, and 2000 mumol/L for 24 h, the growth inhibition rates were (2.02 +/- 2.96)%, (10.82 +/- 2.21)%, (18.95 +/- 3.35)%, (26.88 +/- 2.54)%, and (42.57 +/- 1.27)%, respectively (P < 0.05). SNP altered the cell cycle in AGS cells. Compared with the control group, treatment with SNP at 100, 500, 1000, 1500, and 2000 mumol/L for 24 h reduced the number of cells in the S phase by 2.29%, 7.8%, 11.34%, 20.49%, and 23.6%, respectively, and enhanced the number of cells in the G1/G0 phases by 3.33%, 9.3%, 13.46%, 21.37%, and 24.73%, respectively (P < 0.05). With the increasing concentration and action time of SNP, the expressions of PCNA mRNA and protein decreased. The expression of caspase-3 mRNA remained unchanged, but procaspase-3 was activated.

CONCLUSIONNO not only inhibits cell growth and proliferation, but also induces apoptosis in gastric cancer cells, and such effects of NO showed significant dose-dependent activity.

Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Enzyme Activation ; drug effects ; Gene Expression Regulation, Neoplastic ; Humans ; Nitric Oxide Donors ; pharmacology ; Nitroprusside ; pharmacology ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Stomach Neoplasms ; metabolism ; pathology

OBJECTIVETo investigate the cell viabilities of vascular smooth muscle cells and vascular endothelial cells stimulated by cigarette smoke extract(CSE) .

METHODSThe CSE was prepared by smoke-bubbled phosphate buffered saline(PBS) generation.After culturing cells with different concentrations of CSE, we used the cell counting kit-8 to determine the cell viability.The expression levels of c-jun and cyclinD1 were analyzed through Western blot.The c-jun plasmid was transfected to detect the change of cyclinD1 expression.

RESULTSThe smooth muscle cell viability increased when the CSE concentration ranged 0.625%-10%, whereas the endothelial cells viability decreased when exposed to the CSE concentration. After exposure to CSE for 48 hours, there was no difference in c-jun expression between toxin group and PBS group;however, the expression of p-c-jun in the smooth muscle cells significantly increased in the toxin groups than in the PBS group(P<0.05) and the expression of p-c-jun in the vascular endothelial cells significantly decreased(P<0.05) . The level of cyclinD1 significantly increased after exposed to CSE, and its expression level also increased in respond to the c-jun overexpression.

CONCLUSIONCSE can enhance the proliferation of vascular smooth muscle cells and decrease in the activity of endothelial cells proliferation, which may be explained by the phosphorylation of c-jun and the expression of cyclinD1.

Cell Proliferation ; drug effects ; Cell Survival ; Cells, Cultured ; Cyclin D1 ; metabolism ; Endothelial Cells ; drug effects ; metabolism ; Humans ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Proto-Oncogene Proteins c-jun ; metabolism ; Tobacco ; adverse effects

OBJECTIVETo study the targeted point and mechanism of the function of the blood-activating and stasis-removing Chinese drugs, Paeoniae Radix 801(PR801) in its cardiovascular protective effects and its specific binding with endothelin 1 (ET-1) as well as the dynamics of the two's interactive function by means of using affinity biosensors: IAsys Plus and quartz crystal microbalance (IAQCM).

METHODSET-1 was immobilized on the surfaces of IAQCM by using the new surface modification methods. The PR801 in the solution was detected by modified substrates and the specific binding between PR801 and ET-1 was studied.

RESULTSThe curves went up or down after adding PR801. There is specific binding between PR801 and ET-1. The bound mass were 0.458 ng/mm(2) and 133.54 ng/cm(2), respectively. There exists relatively good stability with these two methods.

CONCLUSIONThe affinity biosensors: IAQCM can be used to study the interaction mechanism between PR801 and ET-1, providing a new way to study the interaction mechanism of TCM. PR801 can bind ET-1 specifically in the experiments. Therefore, ET-1 is another target that PR801 can bind specifically besides thromboxane A(2).

Biosensing Techniques ; standards ; Blood Circulation ; drug effects ; Drugs, Chinese Herbal ; metabolism ; therapeutic use ; Endothelin-1 ; metabolism ; Humans ; Quartz