Proteomic analysis for detecting serum biomarkers related to smoking in humans

Background  Smoking is the leading cause of death in the world. This study focused on the difference of the serum proteomic profiling between healthy smokers and nonsmokers in order to find smoking-specific serum biomarkers.

Methods  Pattern-based proteomic profiling of 100 serum samples (from 50 Chinese male smokers and 50 matched nonsmokers) was performed through magnetic bead fractionation coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis (MALDI-TOF-MS) and resulting data were statistically analyzed by Ciphergen ProteinChip software 3.0.2.

Results  We found 72 serum peaks were significantly different between smokers and nonsmokers (P <0.05). Marker peaks of mass-to-charge ratio (m/z) 3159.13, 7561.03 and 9407.32 were smoking-specific.

Conclusion  The preliminary data suggested that smoking-specific serum biomarkers could be detected in humans.

     

Ibutilide decreases defibrillation threshold by the reduction of activation pattern complexity during ventricular fibrillation in canine hearts

Background  Ibutilide has been commonly used for pharmacologic cardioversion of atrial fibrillation and flutter in clinical settings. The objective of this study was to investigate the effects of ibutilide on the defibrillation threshold (DFT), restitution properties, dispersion of refractoriness and activation patterns during ventricular fibrillation (VF).

Methods  Ibutilide was administrated intravenously in six open-chest beagles. Before and after the drug administration, 20-second episodes of VF were electrically induced and recorded with a 10×10 unipolar electrode plaque sutured on the lateral epicardium of the left ventricle. DFT and VF activation patterns, including type of epicardial activation maps, VF cycle length (VF-CL), conduction velocity, wavelength (WL) and reentry incidence, were measured. Restitution properties and dispersion of refractoriness were estimated from activation recovery intervals (ARI) during pacing.

Results  Compared to baseline, ibutilide markedly decreased the DFT by 31% ((491±14) V vs. (337±59) V, P <0.01). The drug significantly reduced the maximal slope of the restitution curve (1.34±0.08 vs. 0.76±0.06, P <0.01) and its epicardial dispersion (0.36±0.09 vs. 0.21±0.06, coefficient of variation, P=0.03). The dispersion of refractoriness was enhanced at the pacing cycle length of 300 ms to 160 ms by ibutilide. The drug significantly increased the VF-CL ((96±19) ms vs. (112±20) ms, P <0.01) and the WL ((41±9) mm vs. (52±14) mm, P=0.02) during VF, and reduced the reentry incidence by 25% (0.08±0.02 vs. 0.06±0.02, P <0.01). In the epicardial activation maps, ibutilide significantly reduced the percentage of more complex activation maps during VF.

Conclusions  Intravenous ibutilide significantly decreased the DFT. It might be due to reduction of activation pattern complexity during VF.

     

Ibutilide decreases defibrillation threshold by the reduction of activation pattern complexity during ventricular fibrillation in canine hearts

Background  Ibutilide has been commonly used for pharmacologic cardioversion of atrial fibrillation and flutter in clinical settings The objective of this study was to investigate the effects of ibutilide on the defibrillation threshold (DFT), restitution properties, dispersion of refractoriness and activation patterns during ventricular fibrillation (VF).

Methods  Ibutilide was administrated intravenously in six open-chest beagles. Before and after the drug administration, 20-second episodes of VF were electrically induced and recorded with a 10×10 unipolar electrode plaque sutured on the lateral epicardium of the left ventricle. DFT and VF activation patterns, including type of epicardial activation maps, VF cycle length (VF-CL), conduction velocity, wavelength (WL) and reentry incidence, were measured. Restitution properties and dispersion of refractoriness were estimated from activation recovery intervals (ARI) during pacing.

Results  Compared to baseline, ibutilide markedly decreased the DFT by 31% ((491±14) V vs. (337±59) V, P<0.01). The drug significantly reduced the maximal slope of the restitution curve (1.34±0.08 vs. 0.76±0.06, P<0.01) and its epicardial dispersion (0.36±0.09 vs. 0.21±0.06, coefficient of variation, P=0.03). The dispersion of refractoriness was enhanced at the pacing cycle length of 300 ms to 160 ms by ibutilide. The drug significantly increased the VF-CL ((96±19) ms vs. (112±20) ms, P<0.01) and the WL ((41±9) mm vs. (52±14) mm, P=0.02) during VF, and reduced the reentry incidence by 25% (0.08±0.02 vs. 0.06±0.02, P<0.01). In the epicardial activation maps, ibutilide significantly reduced the percentage of more complex activation maps during VF.

Conclusions  Intravenous ibutilide significantly decreased the DFT. It might be due to reduction of activation pattern complexity during VF.

     

Role of microRNA-181a in the apoptosis of tubular epithelial cell induced by cisplatin 

Background  Cisplatin (DDP) is one of most effective and most commonly used therapeutic agent in treating tumors, it can accumulate in the kidney and lead to acute renal failure. MicroRNA-181a can induce cell apoptosis by suppressing the expression of Bcl-2 family. In the present study, we investigated the role of microRNA-181a in the apoptosis of tubular epithelial cell induced by DDP.

Methods  HK-2 cells were cultured, transfected with microRNA-181a inhibitor for 48 hours, and stimulated with 50 µmol/L cisplatin for 24 hours. MicroRNA-181a expression was analyzed by real time PCR, and cell apoptosis was detected by flow cytometry. Moreover, Bcl-2 and Bcl-2-associated X protein (Bax) expression were measured by Western blotting.

Results  MicroRNA-181a expression significantly down-regulated in cells transfected with microRNA-181a inhibitor, compared with that in untransfectd cells (21.19±2.01 vs. 38.87±1.97, P <0.05). Cell apoptosis induced by DDP significantly decreased in cells transfected with MicroRNA-181a inhibitor. Compared with DDP treated cells alone, Bcl-2 expression strikingly was up-regulated and Bax expression was down-regulated in cells transfected with microRNA-181a inhibitor.

Conclusion  One pathway of DDP induces apoptosis of tubular epithelial cell by suppressing Bcl-2 expression is achieved by regulating the target gene of MicroRNA-181a.

     

Serum antibodies to 25 myelin oligodendrocyte glycoprotein epitopes in multiple sclerosis and neuromyelitis optica: clinical value for diagnosis and disease activity

Background  Whether antibody to myelin oligodendrocyte glycoprotein (MOG) can be a diagnostic marker for multiple sclerosis (MS) is still controversial. Recent studies suggested that serum specific anti-MOG epitope antibody might be an MS specific marker. However, these studies did not include neuromyelitis optica (NMO) which might be proven to also have anti-MOG antibody. Hence, the present study was undertaken to investigate the clinical value of serum antibodies to 25 MOG epitopes in conventional MS (CMS) and NMO.

Methods  Serum anti-MOG epitope IgG was detected in 61 CMS patients, 54 NMO patients, and 77 healthy controls, using enzyme-linked immunosorbent assay (ELISA).

Results  Anti-MOG_27-38 IgG levels in both CMS and NMO patients were significantly higher than that in healthy controls (optical density (OD): 0.64±0.38, 0.48±0.23 vs. 0.19±0.09; P=0.000). CMS and NMO patients in relapse stage had significantly higher anti-MOG_27-38 IgG level than patients in remission stage (OD: 0.55±0.14 vs. 0.24±0.09, P=0.027).

Conclusion  Although serum anti-MOG epitope IgG could not differentiate MS from NMO, it may be a useful marker for monitoring disease activity.

     

Antiviral therapy of decompensated hepatitis B virus-related cirrhosis

Objective  To review the development, mechanism, necessity and limitation of antiviral therapy in decompensated hepatitis B virus-related cirrhosis.

Data sources  Most information was pulled from a literature search (Pubmed 2000 to 2011) using the keywords of antiviral and decompensated hepatitis B virus-related cirrhosis. Relevant book chapters were also reviewed.

Study selection  Well-controlled, prospective landmark studies and review articles on antiviral therapy in decompesated hepatitis B virus-related cirrhosis were selected.

Results  Specific antiviral agents not only control viral replication, which permits liver transplantation, but also improve liver function so significantly that patients could be removed from the transplant waiting list. However, the emergence of drug-resistant mutants can result in treatment failure. Combination therapy is a save-strategy in drug-resistant.

Conclusions  Although the treatment of end-stage liver disease is still a challenge worldwide, antiviral therapy has altered the natural history of hepatitis B patients with decompensated cirrhosis. The approval of the new generation of antivirals is opening new perspectives for finding the optimal antiviral treatment for patients with decompensated cirrhosis and preventing antiviral resistance. A combination of antivirals may be one of the future strategies for fulfilling these goals.

     

Anteroapical aneurysm plication improves mechanical intraventricular dyssynchrony in patients with anterior myocardial infarction

Background  Left ventricular (LV) dyssynchrony has been described to occur in patients with myocardial infarction. Dyssynchrony of left ventricular mechanical contraction produces adverse hemodynamic consequences. This study aimed to test the capacity of geometric rebuilding by aneurysm plication to restore a more synchronous contractile pattern after a mechanical, rather than electrical, intervention.

Methods  A total of sixty patients with anterior myocardial infarction, QRS duration <120 ms, electively undergoing operation between January 2008 and January 2010 were included for analysis. Real-time 3-dimensional echocardiography was performed to assess LV function, LV systolic and diastolic dyssynchrony by measuring ejection fraction (EF), peak ejection rate (PER), peak filling rate (PFR) and LV dyssynchrony. LV dyssynchrony was defined as the systolic dyssynchrony of the time to reach the minimum systolic volume for 16 LV segments, expressed in percent cardiac cycle, systolic dyssynchrony index (SDI). We compared changes of LV dyssynchrony at different interval times.

Results  LV contraction was significantly asynchronous because preoperative SDI was higher, EF, PER and PFR were lowered. Compared with function after operation, LV mechanical intraventricular resynchronization was improved with decreased SDI ((8.7±0.5) % vs. (14.3±1.6) %, P=0.01); LV function was improved with EF increasing ((43±9)% vs. (37±7)%, P=0.001), and LV systolic and diastolic dyssynchrony was improved with more rapid PFR (199.4±15.6 vs. 148.4±21.2, P=0.002) and PER (212.4±14.5 vs. 156.3±26.2, P=0.001).

Conclusions  Systolic and diastolic dyssynchrony was highly prevalent in patients with aneurysm, irrespective of QRS duration. Aneurysm plication produces a mechanical intraventricular resynchronization.

     

Transradial artery intervention: an alternative approach for renal artery stent implantation?

Background  Transfemoral artery access is the main approach for the interventional treatment of renal artery stenosis (RAS). This study aimed to investigate the technical feasibility of a transradial interventional (TRI) treatment of renal artery stenosis.

Methods  A series of 23 patients who underwent transradial renal artery stenting from October 2010 to October 2011 were studied. Radial sheath system (Terumo, Japan) was used to get access to the radial artery. Radial tourniquet (Terumo) was used to stop bleeding. A 5Fr MPA (COOK, USA) was used to perform selective renal arteriography. Percutaneous renal artery stent systems were used to perform renal artery stenting. 

Results  Renal artery angiography showed that 15 patients had unilateral renal artery stenosis and eight patients had bilateral renal artery stenosis. The descending aorta could not be catheterized in one patient because of the type III aortic arch. Twenty-two patients successfully underwent transradial renal artery angiography and the technical success rate was 95.7%. There was no puncture site hematoma or pseudoaneurysm. Mean procedure time was (38.4±7.2) minutes, the mean amount of contrast agent used was (93.2±6.3) ml, and the mean postprocedure bleeding time was (3.2±1.9) minutes.

Conclusion  Transradial renal artery intervention is technically reliable with less invasion, rapid recovery, fewer complications and may become an alternative intervention approach for the treatment of renal artery stenosis.

     

Serum major-histocompatibility-complex class I-related chain A antibody detection for the evaluation of graft dysfunction in renal allograft recipients

Background  In addition to the well-known antibodies against human leukocyte antigens (HLA)-induced kidney-graft rejection, polymorphic major-histocompatibility-complex (MHC) class I-related chain A (MICA) antigens can elicit antibodies and have been suggested to play a role in the antibody-mediated allograft rejection (AMR). We carried out a prospective study of MICA antibodies in post-renal transplant patients to determine the association between MICA antibodies, C4d staining, histological features, and graft outcome.

Methods  We tested 52 patients who had biopsy results due to graft dysfunction. The MICA antibodies in concurrent sera were determined by Luminex. All patients were followed up for one year after renal biopsy. The influence of antibody production on the function of graft was analyzed.

Results  Antibodies against MICA were positive in 15 out of the 52 patients (28.9%). The presence of MICA antibodies was associated with renal-allograft deterioration. During one-year follow-up, the estimated glomerular filtration rate (eGFR) decreased (24.0±3.4)% among recipients with anti-MICA antibodies. However, among recipients without anti-MICA antibodies, the eGFR has declined only (8.4±3.0)% (P=0.017). The association between C4d staining, histological features and MICA antibody production was found no significant difference.

Conclusion  Besides anti-HLA antibodies, the presence of post-transplant MICA antibody is associated with poor graft outcome and increases the risk of graft failure.

     

Endothelial progenitor cell down-regulation in a mouse model of Kawasaki disease

Background  Cardiovascular complications of Kawasaki disease (KD) are a common cause of heart disease in pediatric populations. Previous studies have suggested a role for endothelial progenitor cells (EPCs) in coronary artery lesions associated with KD. However, long-term observations of EPCs during the natural progression of this disorder are lacking. Using an experimental model of KD, we aimed to determine whether the coronary artery lesions are associated with down-regulation of EPCs.

Methods  To induce KD, C57BL/6 mice were administered an intraperitoneal injection of Lactobacillus casei cell wall extract (LCWE; phosphate buffered saline used as control vehicle). Study groups included: group A (14 days following LCWE injection), group B (56 days following LCWE injection) and group C (controls). Numbers of circulating EPCs (positively staining for both CD34 and Flk-1 while staining negative for CD45) were evaluated using flow cytometry. Bone marrow mononuclear cells were cultured in vitro to expand EPCs for functional analysis. In vitro EPC proliferation, adhesion and migration were assessed.

Results  The model was shown to exhibit similar coronary artery lesions to KD patients with coronary aneurysms. Numbers of circulating EPCs decreased significantly in the KD models (groups A and B) compared to controls ((0.017±0.008)% vs. (0.028±0.007)%, P <0.05 and (0.016±0.007)% vs. (0.028±0.007)%, P <0.05). Proliferative, adhesive and migratory properties of EPCs were markedly impaired in groups A and B.

Conclusion  Coronary artery lesions in KD occur as a consequence of impaired vascular injury repair, resulting from excess consumption of EPCs together with a functional impairment of bone marrow EPCs and their precursors.

     

Anatomical study of the anterolateral and posteromedial bundles of the posterior cruciate ligament for double-bundle reconstruction using the quadruple bone-tunnel technique

Background  Several techniques have been described for posterior cruciate ligament (PCL) reconstruction. However, double-bundle PCL reconstruction using the quadruple bone-tunnel technique has been seldom reported. The current study investigated this technique, focusing on the anatomy of the femoral and tibial insertions of the anterolateral (AL) and posteromedial (PM) bundles of the PCL.

Methods  Twenty-two fresh, healthy adult cadaveric knees were dissected and measured. The PCL was divided into the AL bundle and PM bundle at the insertion footprint. The insertion footprints of the AL and PM bundles, their location, size, and the clock positions were measured and described.

Results  On the femur, the clock position of the footprint of the AL bundle was 11:21±0:23 (left) or 0:39±0:23 (right), and the PM bundle was 9:50±0:18 (left) or 2:10±0:18 (right), with the knee flexed at 90 degrees. The distances from the center of the femoral insertions of the AL and PM bundles to the anterior cartilage margins of the medial femoral condyle were (7.79±1.22) mm and (8.36±1.63) mm, respectively. On the tibia, the vertical distances from the center of the tibial insertions of the AL and PM bundles to the tibial articular surface were (3.25±1.20) mm and (6.91±1.57) mm, respectively.

Conclusions  These results have led to a better definition of the anatomy of the AL and PM bundle footprint of the PCL. The technique of double-bundle PCL reconstruction using quadruple bone-tunnel is feasible. Application of these data during PCL reconstruction using the quadruple bone-tunnel technique may help optimize knee stability.

     

Effect of chronic intermittent hypoxia on theophylline metabolism in mouse liver

Background Chronic intermittent hypoxia (CIH) has been associated with abnormalities in the liver,which is the most important organ for drug metabolism.This study aimed to investigate the effect of CIH...     

Specific regulator of eosinophil apoptosis: Siglec-8—new hope for bronchial asthma treatment

Objective  It is known that Siglec-8 is selectively expressed on human eosinophils at a high level and mediates eosinophil apoptosis when crosslinked with its antibody. The aim of our review is to elucidate the molecular and biological characteristic of Siglec-8 and then discuss the function and possible mechanisms of Siglec-8 in eosinophils. Thereby, we will expand our understanding to the regulation of eosinophil apoptosis, and provide important clues to the treatment of asthma and other hyper-eosinophilic diseases.

Data sources  Most articles were identified by searching of PubMed online resources using the key term Siglecs.

Study selection  Mainly original milestone articles and critical reviews written by major pioneer investigators in the field were selected.

Results  Siglec-8 is selectively expressed on human eosinophil and can specifically induce eosinophil apoptosis.

Conclusion  The restricted expression of Siglec-8 on human eosinophil and the rapid progress in understanding its role as cell signaling and activation of death receptors have made it an attractive target for treatment of asthma and other hyper-eosinophilic diseases.

     

Azithromycin inhibits neutrophil accumulation in airways by affecting interleukin-17 downstream signals

Background  Azithromycin can reduce neutrophil accumulation in neutrophilic pulmonary diseases. However, the precise mechanism behind this action remains unknown. Our experiment assessed whether azithromycin inhibits neutrophil accumulation in the airways by affecting interleukin-17 (IL-17) downstream signals.

Methods  Mice were pretreated with azithromycin before murine IL-17A (mIL-17) stimulation. After the mIL-17 stimulation, the levels of six neutrophil-mobilizing cytokines were determined by enzyme-linked immunosorbent assay (ELISA) tests in bronchoalveolar lavage (BAL) fluid; IL-6, CXC chemokine ligand-1 (CXCL-1), CXCL-5, macrophage inflammatory protein-2 (MIP-2), granulocyte colony-stimulating factor (G-CSF), and granulocyte macrophage colony-stimulating factor (GM-CSF). The number of neutrophils in BAL fluid were evaluated by cytospin preparations.

Results  (1) Azithromycin pretreatment significantly inhibited both the release of three neutrophil-mobilizing cytokines (MIP-2, CXCL-5 and GM-CSF) and the accumulation of neutrophils in airways caused by mIL-17 stimulation. (2) The levels of three neutrophil-mobilizing cytokines (IL-6, MIP-2 and GM-CSF) were positively correlated with the numbers of neutrophil in BAL fluid.

Conclusions  Azithromycin can inhibit neutrophil accumulation in the airways by affecting IL-17 downstream signals. This finding suggests that macrolide antibiotic application might be useful in prevention of neutrophilic pulmonary diseases characterized by high levels of IL-17.

     

Investigation of long-term implantation of BuMA stent in a porcine coronary model

Background  Stent-based delivery of sirolimus has been shown to reduce neointimal hyperplasia significantly. However, the long-term effect of the polymer is thought to initiate and sustain an inflammatory response and contribute to the occurrence of late complications. Our study aimed to evaluate the efficacy and safety of the BuMA biodegradable drug-coated sirolimus-eluting stent (BSES) for inhibiting neointimal hyperplasia in a porcine coronary model.

Methods  Four types of stents were implanted at random in different coronary arteries of the same pig: BSES (n=24), bare metal stent (BMS) (n=24), biodegradable polymer coated stent without drug (PCS) (n=24) and only poly (n-butyl methacrylate) base layer coated stent (EGS) (n=23). In total, 26 animals underwent successful random placement of 95 oversized stents in the coronary arteries. Coronary angiography was performed after 28 days, 90 days and 240 days of stent implantation. After 14 days, 28 days, 90 days and 240 days, 6 animals at each timepoint were sacrificed for histomorphologic analysis.

Results  The 28-day, 90-day and 240-day results of quantitative coronary angiography (QCA) showed reduction in luminal loss (LL) in the BSES group when compared with the BMS group;  (0.20±0.35) mm vs. (0.82±0.51) mm (P=0.035), (0.20±0.30) mm vs. (0.93±0.51) mm (P=0.013), and (0.18±0.16) mm vs. (0.19±0.24) mm (P=0.889), respectively. By 28-day, 90-day and 240-day histomorphomeric analysis results, there was also a corresponding significant reduction in neointimal tissue proliferation with similar injury scores of BSES compared with the BMS control; average neointimal area (0.90±0.49) mm² vs. (2.16±1.29) mm² (P=0.049), (1.53±0.84) mm² vs. (3.41±1.55) mm² (P=0.026), and (2.43±0.95) mm² vs. (3.12±1.16) mm² (P=0.228), respectively. High magnification histomorphologic examination revealed similar inflammation scores and endothelialization scores in both the BSES and BMS groups.

Conclusions  The BuMA biodegradable drug-coated sirolimus-eluting stents can significantly reduce neointimal hyperplasia and in-stent restenosis. Re-endothelialization of the BuMA stent is as good as that of the BMS in the porcine coronary model due to the reduced inflammation response to the BuMA stent.

     

Rho-associated coiled kinase inhibitor Y-27632 promotes neuronal-like differentiation of adult human adipose tissue-derived stem cells

Background  Y-27632 is a specific inhibitor of Rho-associated coiled kinase (ROCK) and has been shown to promote the survival and induce the differentiation of a variety of cells types. However, the effects of Y-27632 on adult human adipose tissue-derived stem cells (ADSCs) are unclear. This study aimed to investigate the effects of Y-27632 on the neuronal-like differentiation of ADSCs.

Methods  ADSCs were isolated from women undergoing plastic surgery and cultured. ADSCs were treated with different doses of Y-27632 and observed morphological changes under microscope. The expression of nestin, neuron specific enolase (NSE) and microtubule-associated protein-2 (MAP-2) in ADSCs treated with Y-27632 was detected by immunocytochemistry and Western blotting analysis.

Results  Y-27632 had the potency to induce neuronal-like differentiation in ADSCs in a dose-dependent manner. Moreover, the differentiation induced by Y-27632 was recovered upon drug withdraw. ADSCs treated with Y-27632 expressed neuronal markers such as NSE, MAP-2 and nestin while untreated ADSCs did not express these markers.

Conclusion  Selective ROCK inhibitor Y-27632 could potentiate the neuronal-like differentiation of ADSCs, suggesting that Y-27632 could be utilized to induce the differentiation of ADSCs to neurons and facilitate the clinical application of ADSCs in tissue engineering.

     

Steady-state pharmacokinetics of zidovudine in Chinese HIV-infected patients

Background  The pharmacokinetics of zidovudine (AZT) are possibly influenced by weight, age, sex, liver and renal functions, severity of disease, and ethnicity. Currently, little information is available on the steady-state pharmacokinetics of AZT in Chinese HIV-infected patients. The current study aimed to characterize the steady-state pharmacokinetics of AZT in a Chinese set-up.

Methods  Eleven Chinese HIV-infected patients were involved in the steady-state pharmacokinetic study. In total, 300 mg of AZT, as a part of combination therapy, was given to patients, and serial blood samples were collected for 12 hours. The samples were measured by a high-performance liquid chromatography (HPLC) assay, and the results were analyzed by both the non-compartment model and the one-compartment model.

Results  The C_max of AZT in Chinese patients was higher than that in non-Asian patients. The half-life of AZT, analyzed by the non-compartment model (P=0.02), in male patients ((1.02±0.22) hours) was shorter than that of AZT in female patients ((1.55±0.29) hours). The AZT clearance, analyzed by the one-compartment model (P=0.045), in male patients ((262.60±28.13) L/h) was higher than that in female patients ((195.85±60.51) L/h).

Conclusion  The present study provides valuable information for the clinical practice of AZT-based highly active antiretroviral therapy in a Chinese set-up.

     

非肿瘤细胞凋亡引起肿瘤机体血清细胞色素C增高

Background  Neuroblastoma (NB) is one of the most common malignant solid tumors of childhood. It is still not clear whether the apoptosis of tumor cells or the non-tumor cells contributes to the increase of concentration of cytochrome c (Cyt c) in the serum of the cancer patients. The aim of this research was to identify the source of the Cyt c in the serum when the tumor grows up by subcutaneous inoculation of human NB cells into nude mice.

Methods  We subcutaneously inoculated human NB cells (KP-N-NS) into nude mice and collected the sera of tumor-bearing mice (n=14) and control mice (n=25) 4 weeks later in order to screen for and identify differentially expressed proteins in the serum. Differentially expressed proteins in the serum were screened by surface-enhanced laser desorption/ionization-time-of-flight (SELDI-TOF) mass spectrometry.

Results  The relative intensity of a protein having a mass-to-charge ratio (m/z) of 11 609 was 3338.37±3410.85 in the tumor group and 59.84±40.74 in the control group, indicating that the expression level of this protein in the tumor group was 55.8 times higher than that in the control group. Serum proteins were separated and purified by high-performance liquid chromatography (HPLC). Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was performed to produce peptide mass fingerprints (PMFs). Spectrum analysis and a database search revealed that the highly expressed protein (m/z=11 605.4) from the serum of tumor-bearing mice was the mouse Cyt c.

Conclusions  Increased concentration of Cyt c in the serum of tumor-bearing nude mice might be partially attributed to the secretion of this protein by non-tumor cells.

     

Reversal of multidrug resistance in renal cell carcinoma by short hairpin RNA targeting MDR1 gene

Background  Over-expression of P-glycoprotein (P-gp), encoded by the MDR1 gene, confers multidrug resistance (MDR) in renal cell carcinoma (RCC) and is a major reason for unsuccessful chemotherapy. This study aimed to determine the effct of RNA interference (RNAi) on the reversal of MDR in human RCC.

Methods  We designed and selected one short hairpin RNA (shRNA) targeting MDR1 gene, which is stably expressed from integrated plasmid and transfected by lentivirus fluid in human RCC A498 cell.

Results  The MDR1-targeted RNAi resulted in decreased MDR1 gene mRNA level (P <0.001), almost abolished P-gp expression and reversed MDR to different chemotherapy drugs in the RCC A498 cell line.

Conclusion  MDR could be reversed by RNAi in human RCC A498 cell line, which may be used for clinical application in future.