[ABSTRACT]AIM:ToinvestigatetheroleofquercetinintheapoptosisofhumanbreastcancercelllineMCF-7 and the association with Fas/Fas ligand (FasL) pathway.METHODS: The apoptosis model of MCF-7 cells was estab-lished by the induction with quercetin .The morphological characteristics of apoptotic MCF-7 cells were observed under transmission electron microscope .The apoptotic rates and alternation of mitochondrial membrane potential (Δψm ) in the MCF-7 cells were measured by flow cytometry using fluorescein labeled Annexin V-FITC/PI and JC-1, respectively.FasL neutralizing antibody was applied to block the apoptosis .The expression of Fas/FasL on the cells was detected by immuno-fluorescence technique and flow cytometry , respectively.The influence of SB203580 (an inhibitor of p38 MAPK) on the expression of Fas/FasL was also examined by flow cytometry .The protein levels of p 38 MAPK and p-p38 MAPK were de-termined by Western blot .RESULTS: The phenomenon of nuclear condensation and marginalization in the MCF -7 cells treated with quercetin at 80.0 μmol/L for 48 h was observed under transmission electron microscope .Compared with the control cells , theΔψm was decreased by 17.4%, 44.3% and 68.9% in the MCF-7 cells treated with quercetin at 80.0μmol/L for 24 h, 48 h and 72 h, respectively .The apoptotic rates of MCF-7 cells treated with quercetin at 80.0 μmol/L for 24 h, 48 h and 72 h were (10.2 ±3.3)%, (28.9 ±7.5)%and (39.2 ±8.9)%, respectively.However, the apop-totic rates were decreased to (8.2 ±2.8)%, (19.2 ±5.3)% and (22.5 ±6.9)% after the cells were pretreated with FasL neutralizing antibody , respectively .When MCF-7 cells were treated with quercetin for 24 h, 48 h and 72 h, Fas/FasL expression rates were increased in a time-dependent manner , which were largely inhibited by SB203580.The protein level of p38 MAPK was not changed obviously , but the protein level of p-p38 MAPK was significantly increased at 48 h and 72 h.CONCLUSION: Quercetin up-regulates the expression of Fas/FasL on MCF-7 cells, and induces apoptosis via Fas/FasL pathway .Meanwhile , p-p38 MAPK is potentially critical signaling molecule for up-regulating the expression of Fas/FasL.

Objective To investigate the relationship between the 5-lipoxygenase(5-LO) in mouse lung and during infection in the respiratory system.Methods Investigate the expression of 5-LO in the lung of a mouse by immunocytochemistry.Results The intensity of the immune staining against 5-LO antibody was stronger in infected mice than that in non-infected rats.A weaker immuneoreation was observed in the treated group of animals.Conclusion Changes in the expression of 5-LO in pulmonary tisssues could be designed to detect the treatment efficacy during the lung infection.

Objective To discuss the optimalselection of the puncture path in performing CT-guided pericardial drainage,and to evaluate its clinical feasibility and safety.Methods A total of 114 patients with pericardial effusion,who were admitted to authors' hospital during the period from May 2013 to March 2016,were enrolledin this study.The appropriate body position and suitable needle-puncturing route were selected,and CT-guided pericardial drainage with Seldinger'stechnique was performed.Results Successful puncturing and catheter drainage was obtained in all 114 patients,no any serious complication occurred.The time used for manipulation was 18-30 min.Conclusion The use of right puncture path is of great importance for the performance of CT-guided pericardial drainage for pericardial effusion,this technique is highly feasible and safe for relieving the clinical symptoms of pericardial tamponade.

Heteroclitin D (H.D) was successfully isolated from Kadsurae Caulis by using flash chromatography and recrystallized by methanol, 10.2 mg of H.D was obtained from 4.86 g of crude extract, and the purity determined by HPLC was 99.4%. The structure was identified by UV, IR, MS, and NMR analysis. The fast, simple and efficient method can be applied to the preparation of reference substance of H. D.

A high-speed counter-current chromatography (HSCCC) method was successfully developed for the preparative separation and purification of deoxyschizandrin from Schisandrae Sphenantherae Fructus in one step. The purity of deoxyschizandrin was 98.5%, and the structure was identified by MS, UV and NMR. This method was simple, fast, convenient and appropriate to prepare pure compound as reference substances for related research on Schisandrae Sphenantherae Fructus.

Objective To construct the recombinant plasmid containing human filaggrin gene,purify and identify the immunoreactivity of the recombinant protein,and establish the indirect ELISA to detect AFA for diagnosis of RA.Methods The constructed plasmids were transformed into E. Coli Rosettagami(DE3).This fusion protein was purified by NAT chromatography.ELISA coated with the fusion protein Was established to detect the AFA in serum of patients,which included 114 cases of RA,56 cases of SLE,32 cases of OA and 40 cases of normal controls. The correlation between the results of AFA and anti-CCP in RA group were compared. Results 321 bp fragment of filaggrin gene was amplified and the recombinant expression vector pET-28a( + )-filaggrin was constructed. The sequence of filaggrin gene was the same as the sequence reported in the literatures. The Rosetta-gami (DE3) strains of E. Coli with recombinant vector showed high level of filaggrin protein after induction. The SDS-PAGE showed that the plasmid expressed the filaggrin fusion protein with molecule weight of 14 000 Da. The expression protein could be purified by Ni-NAT with activity. The absorbance value of AFA in RA group was 0.473 ±0. 248 while they were 0. 160 0. 088, 0. 121±0. 070, 0.050 0. ±018 in SLE, OA and normal groups respectively. There were significant differences of absorbance values of AFA between RA and SLE, OA, control group (t = 12.004, 14. 464, 18.078, P<0. 01, respectively). The positivities of anti-filaggrin in RA, SLE and OA were 48.2%, 5.4% and 3. 1% respectively. The positivities of AFA were significantly different between RA, OA and normal control groups (x~2 = 67. 088, P < 0. 01). There was positive correlation of results between AFA and anti-CCP antibody (r = 0.42, P < 0. 05 ) . The consistency rate of results between AFA and anti-CCP was 70. 1%. Anti-CCP was negative in 10 out of 114 patients with AFA positive. AFA can be used to diagnose RA with sensitivity of 48. 2% , specificity of 96.9% , positive predictive value of 93. 2% and negative predictive values of 67. 9% . Conclusions The purified human filaggrin fusion protein is successfully purified. The indirect ELISA method based on the recombinant protein shows good sensitivity and specificity. Joint detection with AFA and anti-CCP can improve the positive rate of detection.

Objective To explore crossed cerebellar diaschisis (CCD) in cerebral gliomas with three dimensional arterialspin-labeling (3D ASL) perfusion MRI.Methods The images of 31 patients with cerebral gliomas and 31 normal subjects were retrospectively analyzed.The cerebral blood flow (CBF) were measured with 3D ASL technology,and the asymmetry index (AI) of cerebellar hemispheric CBF value changes were calculated and compared in patients.And the relationship between the AI values of cerebellar hemispheric and the AI values of tumor region in cerebral hemispheric,tumor histological grade and size were investigated.Results Compared with tumor ipsilateral cerebellar hemisphere of supratentorial glioma,the CBF value of contralateral cerebellar hemisphere significantly reduced (t=5.04,P＜0.01),and the AI values of cerebellar hemispheric in patients obviously increased compared to normal subjects (t=4.13,P＜0.01).But there was no significant difference in the AI values of cerebellum between high grade and low grade gliomas (t=1.31,P＞0.05).In cerebral gliomas patients,there was no significant correlation between the AI values of cerebellar hemispheres and the AI values of cerebral hemisphere (tumor:r=-0.28;tumor parenchyma:r=-0.24;tumor plus edema:r=-0.19,all P＞0.05),and tumor size (r=0.18,P＞0.05).Conclusion Cerebral glioma can cause CCD phenomenon,and 3D ASL is able to quantitatively assess the degree of cerebellar hypoperfusion noninvasively.This phenomenon may not be associated with tumor histological grade,size and the AI values of cerebral hemispheric.

Objective To evaluate the value of contrast enhanced MR imaging (CEMRI)for the diagnosis of focal liver lesions with Meta analysis.Methods Relevant English and Chinese language studies were searched on the Pubmed,EMBASE,EBSCO,OV-ID,CNKI,CBM,VIP,WANFANG databases,respectively.Data were calculated with software of STATA 1 1.0 and Meta Disc 1.4. Results 1 7 of 2836 retrieved studies were included,the pooled sensitivity and specificity for CEMRI with 95% confidence interval (95%CI)were 0.85(0.84-0.87)and 0.86(0.84-0.88),respectively,the AUC of SROC was 0.91 6 8.Conclusion CEMRI can be used as one of the primary examination modalities for focal liver lesions with moderate sensitivity and specificity.

DNA barcoding and HPLC specific chromatogram were used to identify three kinds of Plumeria flowers respectively. DNAs extracted from the three Plumeria species were amplified by PCR with universal primers, and the psbA-trnH region was selected. All the amplified products were sequenced and the results were analyzed by MEGA 5.0. Chemometric methods including principal components analysis and hierarchical clustering analysis were conducted on the SAS 9.0 software to demonstrate the variability among samples. In conclusion, the psbA-trnH of all samples were successfully amplified from total DNA and sequenced. These three varieties of Plumeria can be differentiated by the psbA-trnH region and clustered into three groups respectively through building neighbor joining tree, which conformed to their germplasm origins. However, it was hard to distinguish them by HPLC specific chromatograms combined with chemometrics analysis. These indicated that DNA barcoding was a promising and reliable tool for the identification of three kinds of Plumeria flowers compared to HPLC specific chromatogram generally used. It could be treated as a powerful complementary method for traditional authentication, especially for those varieties which are difficult to be identified by conventional chromatography.

Objective To assess the anti-arrhythmic activity and cardioprotective effects of Wenxin Granula, a traditional Chinese formula (consisting of Salviae Miltiorrhizae Radix, Polygonati Rhizoma, Notoginseng Radix et Rhizoma, Nardostachyos Radix et Rhizoma, Angelicae Sinensis Radix, and Succinum), on heart in ischemic-induced myocardial infarction (MI) rats and compare with those of Amiodarone which have been demonstrated in clinic. Methods Rats were randomly divided into Sham-operated (control), Ml + Amiodarone [5 mg/(kg·d)] (MI), and MI + Wenxin Granula [10 mg/(kg·d)] groups and left anterior descending coronary artery was occluded in each group. After left anterior descending for 12 h, standard lead Ⅱ of administration electrocardiogram was recorded in order to analyze the occurrence of arrhythmia. After one month, the size of the infarct area of heart was evaluated by TTC staining method and haemodynamic function was assessed to detect the heart function. Laser scanning confocal microscope and the technique of patch clamp were used to detect the intracellular Ca2+ ([Ca2+]j) and L-type calcium current (ICa-L), respectively. Results Both Wenxin Granula [10 mg/(kg·d)] and Amiodarone [5 mg/(kg·d)] could markedly decrease the incidence of arrhythmia in heart of rats which were subjected to ischemic injury. After one month, Wenxin Granula could significantly decrease mortality to 22.22% and reduce the infarct area (P < 0.05), but Amiodarone did not. The mechanism may involve that Wenxin Granula attenuated [Ca2+]j decreasing in MI rats. Additionally, Wenxin Granula could obviously ameliorate the impaired heart function of MI rats by decreasing the elevated left ventricular end-diastolic pressure and increasing the attenuated maximum change velocity of left ventricular pressure in the isovolumic contraction or relaxation period. On the other hand, electrophysiological experiment results revealed that Wenxin Granula administration one month later also increased the reduced ICa-L density in rat ventricular myocytes in MI rats. The results of LSCM showed that Wenxin Granula could recover the amplitude of [Ca2+]j decreased by heart failure during long term. Conclusion Wenxin Granula could not only inhibit the incidence of arrhythmia but also decrease the mortality, which was accompanied by recovering the amplitude of [Ca2+]j. This protective effect of Wenxin Granula may partially be mediated through changing ICa-L.as well as increasing [Ca2+]j.