Objective Overexpression of multidrug resistance(MDR)is one of the mechanisms that will lead to chemotherapy failure.Of the MDR pathways in tumor cells,P-glycoprotein (P-gp) encoded by MDR genes is one of the key points.99Tcm-methoxyisobutylisonitrile(MIBI) is an imaging agent that can detect overexpression of MDR in tumor cells.The aim of this study was to observe the relationship between 99Tcm.MIBI uptake kinetics and P-gp levels in tumor cells,with or without MDR reversing agents.Methods A totle of 2×106 cells of human myelogeneous leukemia cell line K562 and its resistant subline(K562/D) were incubated with 8 MBq 99Tcm-MIBI respectively.99Tcm-MIBl accumulation and emux at various time inter-vats and the uptake difference with the presence of different cyclosporin A(O.1-O.4 ug/ml)were also ob-served.Comparison of different cell lines or without and with cyclosporin A were performed with the t-test, and the daa of different groups were compared by q-test.Results 99Tcm-MIBI uptake in K562 and K562/D cell line were 1.559±0.529 and 0.107±0.036,99Tcm-MIBI uptake in k562 was flitleon times higher than k562/D.As compared with K562 cell line with no expression of P-gp,significantly increased the 99Tcm-MIBI uptake of K562/D to 106%,148% and 163%after treated with cyclosporin A(0.1,0.2.0.4ug/ml)was ob-served(t=4.35,4.83,5.88,P<0.05).Conclusiom 99Tcm-MIBI uptake can reflect the P-gP level and multidrug-resistance inhibitor may impact 99Tcm-MIBI uptake in P-gP overexpressing cells.

Objective To study the effect of glucocorticoid on serum interleukin(IL)-1,IL-6,transforming growth factor-?_1(TGF-?_1) and tumor necrosis factor alpha(TNF-?) in children with primary nephrotic syndrome(PNS).Methods The levels of cytokines on serum were compared in 3 different patho-type,and IL-1,IL-6,TGF-?_1 and TNF-? in serum were detected using enzyme-linked immunosorbent assay(ELISA) in 25 children with PNS before and after treatment with glucocorticoids.Results Significant difference of IL-1,IL-6,TGF-?_1 and TNF-? were found between the two group before and after the treatment(P0.05).The level of TGF-?_1 in MCD was lo-wer than that in MsPNG,FSGS before and after the treatment(P0.05),and the level of it in FSGS was higher than that in MsPNG after the treatment(P

The laser-induced breakdown spectroscopy (LIBS) was applied to perform a qualitative elementary analysis on four precious Tibetan medicines, i. e. Renqing Mangjue, Renqing Changjue, 25-herb coral pills and 25-herb pearl pills. The specific spectra of the four Tibetan medicines were established. In the experiment, Nd: YAG and 1 064 nm-baseband pulse laser were adopted to collect the spectra. A laser beam focused on the surface of the samples to generate plasma. Its spectral signal was detected by using spectrograph. Based on the National Institute of Standard and Technology (NIST) database, LIBS spectral lines were indentified. The four Tibetan medicines mainly included Ca, Na, K, Mg and other elements and C-N molecular band. Specifically, Fe was detected in Renqing Changjue and 25-herb pearl pills; heavy mental elements Hg and Cu were shown in Renqing Mangjue and Renqing Changjue; Ag was found in Renqing Changjue. The results demonstrated that LIBS is a reliable and rapid multi-element analysis on the four Tibetan medicines. With Real-time, rapid and nondestructive advantages, LIBS has a wide application prospect in the element analysis on ethnic medicines.
Calcium ; analysis ; Copper ; analysis ; Iron ; analysis ; Lasers ; Medicine, Tibetan Traditional ; Mercury ; analysis ; Silver ; analysis ; Spectrum Analysis ; methods

The study reports the detection of neuroprotective effect of 10 kinds of coumarin derivatives and explores their possible mechanism. MTT method was used to screen the neuroprotective effect of 10 coumarin derivatives on neurotoxic agents (Aβ25-35 and rotenone) or OGD (oxygen-glucose deprivation). A compound with better protective effect was obtained. Then the effect of this compound on neurotoxic agents on PC12 was detected by the morphological observation. Furthermore, the effect of compound 3 on microglia with lipopolysaccharide (LPS) induced inflammation was detected. And the inflammatory factor was tested. Finally, direct free radical scavenging ability was detected. Compound 3 was found to be the best compound through three neurons toxic models. Not only compound 3 ameliorated cell viability reduced by three neurons toxic models, but also significantly inhibited the production of inflammatory factor (TNF-α and IL-1β). And its free radical scavenging ability is very good, especially the effect on superoxide anion, which is comparable with vitamin C. The significant scavenging effect of compound 3 on superoxide anion might be the mechanism of the neuroprotection. Compound 3 as a potential neural cell protective agent merits further investigation.
Animals ; Coumarins ; chemistry ; Free Radical Scavengers ; chemistry ; Inflammation ; Microglia ; drug effects ; Neurons ; drug effects ; Neuroprotective Agents ; chemistry ; PC12 Cells ; Rats

Objective To investigate the clinical role of central venous pressure(CVP) to evaluate fluid responsiveness in septic shock patients. Methods 66 septic shock patients were studied, every patient was administered a volume challenge, before and after it, CVP, intrathoracic blood volume index (ITBVI),global end-diastolic volume index(GEDVI), cardiac index(CI), stroke volume index(SVI) were measured by PiCCO method. All the obtained values were analyzed by statistics method. Results Initial CVP in responders is significantly different from that in nonresponders; △ITBVI, △GEDVI, △CI, △SVI, △HR (△:changes) before and after volume challenge in responders were significantly different from those in nonresponders; the significance of △ITBVI, AGEDVI to predict volume responsiveness was strong indicated by high values of areas under the receiver operating characteristic curves (0.674 and 0.700, respectively).If patients were regrouped by CVP≤11 mm Hg(1 mm Hg=0.133 kPa) and CVP > 11 mm Hg, initial ITBVI and GEDVI in responders were not significantly different from that in nonresponders; △ITBVI,△GEDVI, △CI, △SVI before and after volume challenge in responders were significantly different from those in nom'esponders. Conclusion In septic shock patients, CVP play a guidance role to predict and evaluate volume responsiveness and when CVP was > 11 nun Hg, a positive response will be less likely. Initial volumetric parameters(intrathoracic blood volume and global end-diastolic volume) play a questionable role in predicting and evaluating volume responsiveness, changes before and after volume challenge maybe helpful.

Aim:To develop a rapid high-performance liquid chromatography-mass spectrometry (HPLC-MS) method for the quantification of tiopronin in human plasma.Methods:Cysteine was chosen as antioxidant and firstly added into the whole blood firstly.After adding mycophenolic acid as internal standard (IS) and 1 mol/L HCl into the plasma,the samples were extracted with acetic ether and then determined by HPLC-MS.The chromatographic separation was performed on a Shim-pack VP-ODS C18 column (250 mm×2.0 mm,5 μm) using methanol-0.1% acetic acid (70∶30) as mobile phase with a flow rate of 0.2 mL/min.Results:The assay exhibited a linear range of tiopronin between 30～4 000 ng/mL.The precisions for intra- and inter-batch were all within 8.5%.The extraction recoveries were more than 70%.The total HPLC-MS analysis time was within 7.5 min per a run.The fully validated method was successfully applied to quantify tiopronin in human plasma for a bioavailability study.Conclusion:The assay proved to be accurate,sensitive,selective and convenient.The fully validated method can be applied to study the pharmacokinetics and bioavailability of tiopronin and tiopronin formulations in human.

Objectlve To research and analyze the hemod)rnamic status of refractory septic shock associated cardiac dysfunction.Methods 70 refractory septic shock patients were studied.In the duration of pulmonary artery catheter(PAC)-directed hemodynamic optimization,the patients were divided into a cardiac dysfunction group and a control group.Hemodynamic parameters,arterial blood lactate concentration and APACHE II scores were obtained instantly after the placement of a PAC,then lactate clearance in 24 hours was surveyed and calculated.Subsequently the two groups of patients were regrouped by nonsurvivor and survivors respectively.All the obtained values were analyzed with statistic methods.Results 37% of the refractory septic shock patients was complicated with cardiac dysfunction.The age of the patients complicated with cardiac dysfunction was significantly higher than that of the patients of the control group.Central venous pressure(CVP),pulmonary artery obstruction pressure(PAOP),pulmonary artery pressure (PAP),systemic vascular resistance index(SVRI),pulmonary vascular resistance index(PVRI)and oxygen extraction ratio(O2ext)in the cardiac dysfunction group were significantly different from those in the control group.Cardiac output(CO),cardiac index(CI),oxygen delivery index(DO2I)and mixed venous oxygensaturation(S-v O2)were significantly lower than those of the patients in the control group.S -v O2 had a strong correlation witIl CI.If the patients were regrouped by nonsurvivors and survivors.in the patients complicated with cardiac dysfunction APACHE II scores were significantly higher in the nonsurvivors than survivors:the lactate clearance in 24 hours(median-25%)of the nonsurvivors was significantly lower than that of nonresponders(median 22%),P＜0.05.Conclusion (1)In refractory septic shock patients,cardiac dysfunction maybe the main reason leading to bad outcome.(2)Higher CVP and PAOP and lower S -v O2 indicate the onset of cardiac dysfunction.(3)The patients with significantly high initial arterial blood lactate level and the low lactate clearance in 24 hours had bad outcome.

Objective To observe the expression of Neuropilin (NRP)-1, 2, to calculate micro vessel density (MVD) in colorectal cancer, paraneoplastic tissues, normal tissues, and to investigate the correlation between NRP-1, 2 and MVD to understand the role of NRP-1, 2 in the process of tumor angiogenesis. Methods Expression of NRP-1 and NRP-2 were studied by immunohistochemistry in 66 specimens from colorectal cancer, paraneoplastic tissues and normal tissues. MVD was assessed based on CD105 immunohistochical staining. Results (1) The positive expression of NRP-1, 2 in colorectal cancer, paraneoplastic tissues, normal tissues were 71.2%, 25.8%, 0;80.3%, 15.2%, 0 respectively. There was a statistical difference between them (P<0.05). (2)The quantity of MVD in colorectal cancer, paraneoplastic tissues and normal tissues were 35.682±5.542, 14.485±3.301 and 6.864±1.771 respectively. There was a statistical difference between them (P<0.05). (3) Expression of NRP-1, 2 had a good correlation with the tumor size, infiltrating depth, lymph node metastasis, Dukes stag-ing (P<0.05), but no correlation was found with the tumor position, histological types, differentiation degree and the gender or age of patients (P>0.05). (4) The MVD value had no correlation with the gender or age of patients, tumor size, position, histological types (P>0.05), but had a good correlation with infiltrating depth, lymphatic metastasis and Dukes staging (P<0.05). (5) There was a positive correlation between NRP-1, 2 and MVD value (P<0.01). Conclusion NRP-1, 2 may play an important role in the angiogenesis of the colorectal cancer, and it correlates with the invasion and metastasis of colorectal cancer closely.

Background Allograft rejection is a main cause of failure of penetrating keratoplasty,especially in the patient with high risk of rejection condition.Previous study on allograft rejection mechanism focused on limbal and corneal neovascularization,but these factors did not explain all the phenomena of allograft rejection.Research found that immune cells appeared in iris and ciliary body when rejection occurred,but the relationship between these immune cells and allograft rejection is unclear Objective This study was to evaluate the relationship between diversity of vascular permeability in the iris and ciliary body and allograft rejection after penetrating keratoplasty.Methods Seventy clean eight-week-old BALB/c mice were divided into allogeneic corneal transplantation group (60 mice) and blank control group (10 mice).Allogeneic corneal transplantation was performed with the same age of C57BL/6 mice as donor and BALB/c mice as the recipients.The grafts were examined under the slit lamp microscope and scored based on the criteria of Hegde.The mice were sacrificed and iris and ciliary tissue were obtained 5,10 days and rejection after surgery.Immunohistochemistry and reverse transcription PCR (RT-PCR) was used respectively to detect the expression diversities of occludin,zonula occludens protein-1 (ZO-1),matrix metalloproteinase-9(MMP-9),major histocompatibility complex-Ⅱ (MHC-Ⅱ),and CCR5,CCR7 and their mRNA in iris and ciliary body.Image-J image analysis software was used to calculate the quantity of positive cells on iris wholemount,and absorbance of target genes (A values).The use and care of the experimental animals complied the ARVO Resolution on the Use of Animals in Research.Results The mean survival time of corneal gratts was (17±3) days after operation.The mean score was 0.6 in 5 days and 0.5 in 10 days,and 3.3 in 18 days after operation.Expression of ZO-1 reduced significantly,and that of MMP-9 increased obviously at the time of rejection.MHC Ⅱ + cells were scattered in iris and ciliary body in normal mice,and the number of the positive cells (cells/field) was increased after operation with a peak value when rejection occurred.A significant difference was seen between normal mice and rejection mice (1559.67±350.29 vs.4021.83±495.18) (P=0.000).The expressions of occludin mRNA and ZO-1 mRNA in the iris and ciliary body decreased obviously in the rejection mice.Compared with normal mice,theA value of ZO-1 and occluding were 36.74±3.13 vs.110.11±11.88 and 57.54±3.41 vs.59.90±3.50respectively,with significant differences between them (all P＜0.05).The expressions of MMP-9 mRNA,CCR5 mRNA and CCR7 mRNA in the iris and ciliary body increased gradually with the time lapse after operation and peaked when the rejection appeared.The A value of MMP-9 mRNA,CCR5 mRNA and CCR7 mRNA were significantly higher than those of normal mice (20.29±1.19 vs.2.77±0.85 for MMP-9 mRNA; 35.43±2.56 vs.9.11±0.29 for CCR5 mRNA,and 60.83±0.87 vs.0.89 ±0.95 for CCR7 mRNA) respectively (all P＜0.05).Conclusions The permeability of vascules in the iris and ciliary body increase during the allograft rejection after penetrating keratoplasty.Increased antigen presenting cells were also detected.

Objective To compare the heart function and observe the clinical prognosis of dilated cardiomyopathy(DCM) in children treated with combination of prednisone and azathioprine and high dose intravenous immunoglobulin(IVIG).Methods From Jan.1998 to May 2004,20 children who met the diagnostic criteria of DCM were divided into 2 therapeutic groups(groupⅠ,groupⅡ) according to the time of hospitalization.On the basis of traditional congestive heart failure(CHF) therapy,the children in groupⅠwere treated with immunosuppressive therapy(prednisone plus azathioprine) and the children in groupⅡ were treated with high dose IVIG.The course of treatment was 6 months for all children.Heart functions in all children were evaluated and echocardiographic parameters including left auricular diameter(LA),left ventricular end diastolic diameter(LV),left ventricular ejection fraction(LVEF) and cardiac index(CI) were measured before and after therapy,respectively.All children were followed up for 1 year to observe survival rate(%).The findings were analyzed by t test and ?~2 test.Results The percentage of heart function increased from Ⅲ-Ⅳ toⅠ-Ⅱin groupⅠ,which was lower than that in groupⅡ(P