Objective To investigate mechanisms of resistance to adriamycin(ADR)by human breast cancer cell line MCF-7 and to find the alteration of features and celluar behavior of MCF-7 after exposure to ADR.MethodsProliferation speed,population doubling time of MCF(wild type),MCF-7/ADR(exposure to adriamycin)and withdrawl group were respectively tested.Cell phenotype alteration was detected using SP immunohiatochemistry methods.Results No significant difference of proliferation speed was found between MCF-7/ADR and MCF cells.As exposure time prolonged,withdrawl group cells grew faster,thus population doubling time shortened.Differentiation of MCF-7/ADR and wthdrawl group was lower than wild group.The expression of drug resistance associated marker of MCF-7/ADR such as Pgp,LRP,GST-pi,TOPOⅡwas higher than that of MCF-7,ER turned to express negatively,and expression of PR gradually decreased as exposure continued.Conclusion MCF-7 cells exposed to ADR got drug resistant,their proliferation was not suppressed by withdrawl of ADR and even grew faster.Drug resistant cells gained dedifferentiation ability.Their heredity and biochemistry features changed,expression of target enzyme also altered and was reversible by drug withdrawl.

Objective To investigate the protective function and mechanism of microRNA-1 (miR-1) downregula-tion in H2O2 injured cardiomyocytes. Methods The experiment was divided into 4 groups:Blank group, Negative Control group (NC), H2O2 group and H2O2+AS-miR-1 group. Cells in blank group were not underwent any treatment. Cells in NC group were transfected with random miRNA fragment. H2O2 and H2O2+AS-miR-1 groups were defined as H2O2 injured car-diomyocytes without or with transfected antisense miR-1 oligonucleotide(AS-miR-1). Real time PCR was used to test miR-1 transcription level, and cell vitality and apoptosis were analyzed by MTT and flow cytometry. The target gene of miR-1 was predicted by bioinformatics, then its mRNA transcription and protein expression level of Bcl-2 were detected by real time PCR and western blot respectively. Results There is no significant difference of all index between Blank group and NC group. H2O2 can induce cardiomyocyte injury, increase miR-1 level and rise apoptosis rate, reduce cell vitality and de-crease Bcl-2 expression level. Transfection of AS-miR-1 can decrease cell apoptosis, increase cell vitality and enhance Bcl-2 expression level. Conclusion Downregulation of microRNA-1 can protect cardiomyocytes that was injured by H2O2 through increasing anti-apoptosis factor Bcl-2 expression.

The specific fragment of Pneumococcal surface protein A(PspA)and Pneumococcal Surface Adhesin A(PsaA)gene was amplified by PCR from Streptococcus pneumonia 5 and Streptococcus pneumonia 19.The amplified fragnent of PspA and PsaA gene was ligated into pET-27b(+)vector and transformed into BL 21 E.coli for expression and obtain the expressive production of PspA and PsaA.Induced by IPTG,the expression level was as high as 75 % of the total disolube protein.The result showed that the recombinant plasmid could express a specific 75 kDa and 37 kDa fusion protein in E.coli BL 21,which showed the good immunogenicity and a broadly cross reactivity with the other serotypes.

In this study, the optical data of tongue color of different syndromes in primary hepatic carcinoma (PHC) were detected by optical spectrum colorimetry, and the chromaticity of tongue color was compared and analyzed. The tongue color characteristics of different syndromes in PHC and the relationship between different syndromes and tongue color were also investigated.

OBJECTIVETo study the mechanism of reversal effect of Curcuma Wenyujin n-Butyl alcohol extract (CWNAE) on multiple drugs resistance (MDR) of SGC7901/VCR cells.

METHODSSGC7901/VCR cells were co-culured with different concentrations CWNAE (80, 40, and 20 μg/mL) and Verapamil (VP, 10 μg/mL) for 24 h, and then acted with Adriamycin (ADM) for 1, 2, and 4 h, respec- tively. SGC7901/VCR cells with no intervention were taken as the vehicle control group. SGC7901/VCR cells treated with ADM alone were taken as the control group. The effect of CWNAE on intracellular ADM concentration was detected by flow cytometry (FCM). Cells were treated as mentioned before without any intervention of ADM. SGC7901/VCR with no ADM intervention were taken as the control group. The effect of CWNAE on the expression of P-glycoprotein (P-gp), lung resistance protein (LRP), and glu- cosylceramide synthase (GCS) was studied by Western blot. The effect of CWNAE on the location and expression quantity of P-gp was further illustrated by immunohistochemistry (IHC).

RESULTSCompared with the ADM group, the expression ratio obviously increased in the W80, W40, W20, and VP10 groups with statistical difference (all P < 0.05). The comparative expression quantity of P-gp, GCS, and LRP in SGC7901/VCR cells was obviously higher than that of non-MDR with statistical difference (all P < 0.05). The expression quantity of P-gp and GCS could be obviously down-regulated by 80 and 40 μg/mL CWN- AE, and 10 μg/mL VP, with no effect on the expression of LRP. Results of IHC proved that P-gp was mainly expressed on the cytomembrane or in the plasma, and it was also expressed on the nuclear membrane. P-gp in different locations could all be down-regulated by CWNAE.

CONCLUSIONSCWNAE could reverse the MDR of SGC7901/VCR cell line probably by inhibiting the expression of P-gp and GCS. CWNAE had no effect on LRP that also highly expressed on SGC7901/VCR. So we supposed that CWNAE could become a potential drug to reverse MDR of highly expressed P-gp and GCS.

ATP-Binding Cassette, Sub-Family B, Member 1 ; Cell Line, Tumor ; Curcuma ; Doxorubicin ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Stomach Neoplasms

Objective To investigate the effects of different degrees of hemodilution with the artificial plasma substitutes most commonly used in clinical practice on coagulation in vitro. Methods Ten male ASA physical status I volunteers aged (28.8?1.6) yr and weighing (66?8) kg were enrolled. Venous blood samples obtained from each volunteer were diluted with 10% HES (200 000/0.4-0.55), 4 % succinylated gelatin (gelofusine GEL), 3.5 % polygeline (Haemaccel HAE) and lactated Ringer's solution (RL) by 33% [blood: diluent(B:D) =2:1], 50% (B:D=1:1) and 66% (B:D=1:2). Coagulation of the undiluted blood (control) and diluted blood was measured with sonoclot coagulation and platelet function analyzer (SCT) and by routine coagulation tests. The parameters measured included activated clotting time (ACT), clot rate, platelet function (PF), Hct, platelet count (Pit), plasma fibrinogen concentration (Fig) and activated partial thromboplastin time (APTT) . Results (1) At 33 % dilution ACT was significantly shortened in all the four groups as compared with control value; at 50 % dilution ACT was significantly shorter than the control in RL, HES and HAE groups; at 66 % dilution ACT was significantly prolonged in HES group. (2) Clot rate was significantly decreased at 33 % dilution only in HES group but was significantly decreased in all the four groups at 50 % and 66 % dilution. (3) PF decreased significantly only in GEL group at 33 % dilution but was significantly decreased in GEL and HES groups at 50 % and 66 % dilution. (4) With increasing dilution Hct, Pit and Fig gradually decreased and APTT was prolonged. Conclusion Coagulation changes are closely related to the degrees of dilution. At 33 % dilution, the three artificial plasma substitutes tested do not significantly affect hemostasis. At 50 % and 66 % dilution coagulation is badly impaired, but there are differences among the substitutes used for dilution. HAE impairs oagulation least as it contains higher calcium

Objective To investigate the expression level of adenosine 2A receptors in peripheral blood lymphocytes and the correlation with disease progression of Parkinson's disease (PD).Methods In this retrospcctive study,out patients with PD(42 cases)and healthy controls(32 cases) were recruited at Beijing Hospital.The expression level of A2A receptors in peripheral blood lymphocytes was detected by flow cytometry.The concentration of free A2A receptors in plasma was detected by enzyme linked immunosorbent assay (ELISA).The expression of A2A receptors and plasma free A2A receptors in peripheral blood lymphocytes of the PD group and the control group was compared.Multivariate regression analysis and single factor correlation analysis were performed on sex,age,course of disease,duration of medication,drug type and dose,motor complications,and PD unified Parkinson's disease rating scale (UPDRS)score.Results A2A receptor expression in lymphocytes was significantly higher in the PD group than in the control group (8.96 ± 4.73)％ vs.(5.39±2.42)％ (t=4.210,P＜0.05).There was no significant difference in A2A receptor concentrations in plasma between the two groups (1.82 ± 1.91) μg/L vs.(1.15 ± 0.71) μg/L(t=1.078,P＞0.05).In the PD group,A2A receptor expression in lymphocytes in patients with motor complications was statistically lower than in patients without them (P＜ 0.05).Lymphocyte A2A receptor levels in the 5-9 years duration subgroup were significantly lower than those in the ＜5 years duration subgroup (Z=2.780,P＜0.01) and the≥10 years duration subgroup (Z=-2.149,P＜0.05).Conclusions The expression of A2A receptors in peripheral blood lymphocytes is correlated with PD.The expression of A2 A receptors in peripheral blood lymphocytes of patients with PD fluctuates with the occurrence of motor complications and the progression of disease.Further research is needed to establish A2A receptors as a biomarker for monitoring disease progression.

AIM: To investigate the effect of Qingrehuayuziyin Fang on lymphocyte apoptosis and expression of Fas and FasL in peripheral blood in the patients with systemic lupus erythematosus(SLE).METHODS: Eighty patients with SLE were selected and divided randomly into two groups,40 in the therapy group and 40 in control group respectively.Patients were get the treatment of Qingrehuayuziyin Fang combined hormone(prednisone) as therapy group and get hormone treating only as control group.After treating three months,lymphocyte were collected to be detected the rate of apoptosis by flow cytometer.Meanwhile,the expression of Fas and FasL were analyzed by RT-PCR.RESULTS: After three months treating,the rate of lymphocyte apoptosis was decreased significantly in therapy group compared with that in control group(P

Objective To investigate the prevalence of Helicobacter pylori (HP) infection in patients with Parkinson's disease (PD), and analyze the relationship of HP infection with the symptoms, disease progression and motor complications of PD.Methods 72 PD outpatients were randomly selected, and 100 gender and age-matched subjects who took health physical examination were considered as healthy controls.Basic data were collected and estimated by Unified Parkinson's Disease Rating Scale (UPDRS), Hoehn and Yahr stage, the 39-item Parkinson' s Disease Questionnaire (PDQ-39), Activity of Daily Living Scale (ADL), Minimum Mental State Examination (MMSE), Hamilton Depression Scale (HAMD), Hamilton Anxiety Scale (HAMA).All subjects underwent 13C-urea breath test to determine Helicobacter pylori infection status.Results There was no significant difference in H P infection rate between PD and control groups [37.5 ％ (27 cases)vs.32.0％ (32 cases), P=0.45].According to the initial symptoms, PD patients were divided into two groups: bradykinesia/ rigidity group and tremor group.The HP infection rate was higher in bradykinesia/rigidity group than in tremor group [52.2％ (12 cases) vs.30.6％ (15 cases)], but had no significant difference (P=0.08).The mean HY stage was higher in the HP-infected patient group than in the non-infected group [(2.6±0.7) vs.(2.2±0.9), P=0.03].The HP infection rate was higher in patients with HY stage ≥2 than with HY stage ＜2 [46.4％ (26 cases) vs.6.3％ (1 case),P=0.003].There were no correlations of HP infection with gender, age, age of onset, UPDRS, UPDRS-Ⅲ, PDQ-39, the course of the disease, wearing-off phenomenon, and dyskinesias.Conclusions The PD patients with HP infection are mainly at middle-and late-stage.HP infection may be related with the progress of the disease.It needs further study and validation by expanding the sample to investigate whether HP infection affects the motor complications.

Objective To evaluate the effect of Parkinson's disease treatment guideline on the initial medicine therapy and to explore the impact of hospital grade level and type of insurance on the initial prescription for Parkinson's disease.Methods We identified 136 PD patients as part of a population-based study in Beijing and made a comparison between the patients with initial prescriptions of dopamine receptor agonists (DA) and levodopa (LD).Results Among 136 patients,excluding patients without initial medication of levodopa or dopamine receptor agonist,there were 61 cases with initial drug administration aged ≥ 65 years.Among the 61 cases,1 cases and 19 cases administrated dopamine receptor agonist and levodopa respectively before 2006,1 case and 11 cases respectively from 2007 to 2009,and 5 cases and 24 cases respectively after 2010.There was no significant difference in the percentage between patients with initial administration of levodopa and dopamine receptor agonist in the three periods (x2 =1.891,P =0.388).In 41 patients with initial drug administration of dopamine receptor agonist or levodopa aged＜65 years,1 cases and 23 cases administrated dopamine receptor agonist or levodopa respectively before 2006,0 case and 10 cases respectively from 2007 to 2009,and 3 cases and 4 cases respectively after 2010.There was significant differences in the percentage between patients with initial administration of levodopa and dopamine receptor agonist in the three periods (x2 =10.644,P=0.005).The percentage of patients with initial administration of dopamine receptor agonist aged＜65 years was increased after 2010 as compared before 2006 (x2 =7.219,P=0.028).There were no significant differences in the percentage of patients with initial administration of dopamine receptor agonist between grade 3 and non-grade 3 hospitals (13.6％ vs.6.3％,x2=0.675,P=0.686)and between patients with and without insurance (13.1％ vs.10.0％,x2=0.141,P=1.000).Conclusions The percentage of patients with initial administration of levodopa and dopamine receptor agonist has no significant difference between patients with initial drug administration aged ≥ 65 years before and after the introduction of Parkinson's disease treatment guideline,while the percentage is increased in patients with initial drug administration aged＜65 years.No impact of hospital grade level and type of insurance on initial administration for Parkinson's disease is found.