Objective:To evaluate the benefit of autologous stem cell transplantation (ASCT) as a consolidation therapy in the survival of multiple myeloma (MM) patients at different risks. Methods:A total of 67 MM patients who received ASCT as consolida-tion therapy between August 2006 and July 2011 were enrolled in the retrospective study. The cases were divided into three risk groups on the basis of the International Staging System and fluorescence in situ hybridization. Another 67 patients who accepted consolidation chemotherapy at the same period were selected as case-paired controls matched in terms of age, sex, optimal response after induction, and risk stratifications. All the patients received bortezomib-and/or thalidomide-based induction therapies. Results:No statistical differ-ences in non-complete remission (nCR)/complete remission (CR) rate were observed between the ASCT and chemotherapy groups (44.8%vs. 37.3%, P=0.380) after the induction therapy. The progression-free survival (PFS) was longer in the ASCT group than in the chemotherapy group (32.4 months vs. 15.1 months, P<0.001). The overall survival (OS) was longer in the ASCT group than in the che-motherapy group (58.8 months vs. 42.1 months, P=0.009). both the PFS (median:30.5 months vs. 11.2 months, P<0.001) and the OS (median:85.5 months vs. 34 months, P=0.015) rates were significantly prolonged in the high-risk subgroup after ASCT. In the interme-diate-risk subgroup, neither PFS nor OS showed any significance after ASCT (P>0.05). In the low-risk subgroup, only PFS was extend-ed (median: 34.8 months vs. 17.6 months, P=0.012) after ASCT, without significant improvements in the OS (P>0.05). Conclusion:The MM patients obtained cytogenetic high-risk benefits mostly from ASCT consolidation after inductions based on novel agents.

BACKGROUND:HLA-DRB1 is related to the pathogenesis of alergic rhinitis. Construction ofHLA-DRB1 gene knockout animal models not only elucidates the pathogenesis of alergic rhinitis, but also provides a good way for the elucidation of the pathogenesis of alergic rhinitis-related diseases. OBJECTIVE:To establish the HLA-DRB1gene knockout animal models. METHODS:Homozygous, wild-type and heterozygous mice were obtained by inbreeding of the heterozygous mice. Confirmed by gene and protein identification, 24 female wild-type (H2-eb1+/+) mice and 12 H2-eb1-/-mice aged 8 weeks were selected according to the random number table. 12 H2-eb1+/+ mice and 12 H2-eb1-/- mice were sensitized with ovalbumin to establish the mouse models of alergic rhinitis. Another 12 mice were sensitized with PBS as comparison. RESULTS AND CONCLUSION:Compared with the control mice, serum levels of ovalbumin-specific IgE and interleukin-4 were significantly increased, while serum level ofγ-interferon was significantly decreased in the mouse models of alergic rhinitis. Serum levels of IgE and interleukin-4 were lower, while serumγ- interferon level was higher, inH2-eb1-/-gene knockout mice of alergic rhinitis than those in the H2-eb1+/+ gene knockout wild-type mice. These results suggest thatH2-eb1 gene may play an important role in regulating Th1/Th2 imbalance in the pathogenesis of alergic rhinitis.

Objective To investigate the effects of HMGB1 small interference RNA (siRNA) on retinal vascular endothelial cells.Methods siRNA was used to inhibit the expression of HMGB1,followed by the application of CCK8 assay,Hochest33342 staining and flow cytometry to observe the effects of HMGB1 siRNA on retinal vascular endothelial cells in high glucose environment.Meanwhile,the expression of proteins related to apoptosis was detected by Western blot.Results The transfection of HMGB1 siRNA down-regulated the protein expression level of HMGB1 by 73％ in siRNA group compared with normal control (NC) group (P ＜ 0.05),and the protein expression level of HMGB1 in siRNA group was decreased by 75％ compared with scr-siRNA group (P ＜ 0.05).There was no significant difference between NC group and scrsiRNA group (P ＞ 0.05).The total apoptotic rate of NC group,high-glucose group,scrsiRNA group and siRNA group was (0.40 ± 0.03)％,(49.80 ± 3.50)％,(47.60 ±1.98) ％ and (23.60 ± 2.40) ％ by flow cytometry.Compared with NC group,the apoptotic rates of high-glucose group,scr-siRNA group and siRNA group were increased (all P ＜ 0.05).Compared with scr-siRNA group,the apoptotic rate of HRECs in siRNA group was reduced,with significant statistical difference (P ＜ 0.05).There was no significant difference in the rate of cell apoptosis between scr-siRNA group and high-glucose group (P ＞ 0.05).Compared with the NC group,the protein expression level of cleavedcaspase3 protein in high-glucose group and scr-siRNA group were increased by (233 ±10) ％ and (266 ± 22) ％,respectively (both P ＜ 0.05);compared with scr-siRNA group,the protein expression level of cleaved-caspase 3 in siRNA group was reduced by (43 ±3) ％ (P ＜ 0.05);and there was no significant difference in the protein expression of cleaved-caspase 3 in high-glucose group and scr-siRNA group (P ＞ 0.05).Compared with the NC group,the protein expression of B-cell lymphoma-2 (Bcl-2) in high-glucose group and scr-siRNA group was decreased by (32 ± 2) ％ and (29 ± 3) ％,accordingly (both P ＜ 0.05);compared with scr-siRNA group,the protein expression level of Bcl-2 in siRNA group was increased by (42 ± 2) ％ (P ＜ 0.05);and there was no significant difference in the protein expression of Bcl-2 in high-glucose group and scr-siRNA group (P ＞ 0.05).Conclusion HMGB1 siRNA can reduce the apoptosis of retinal vascular endothelial cells in high glucose environment by inhibiting the activation of cleavedcaspase3 and increasing the expression of Bcl-2.

OBJECTIVETo observe the proliferation state of transplanted cells in acute myocardial infarction (AMI) rats, and the endothelial progenitor cells (EPCs) preconditioned by salvianolic acid B in different ratios with the bone mesenchymal stem cells (BMSCs).

METHODSThe cultivation and purification of EPCs were performed by density-gradient centrifugation and plastic adherence method. Two types of cells were identified by immunocytochemical method (CD34, CD133, and CD44). The rat model of AMI was prepared by ligation of left anterior descending artery. The EPCs were pre-treated with the optimal concentration of salvianolic acid B (8 microg/ mL). They were mixed with BMSCs in different proportions (EPCs/BMSCs in the ratio of 1:1, 2:1, 4:1, and 8:1, respectively). BMSCs and EPCs were injected into the myocardial infarction area. The infarcted area was determined by the N-BT staining and hematoxylin-eosin staining. The expression of Ki-67 was detected by immunohistochemical assay.

RESULTSCompared with the model group (19.60% +/- 3.23%), the myocardial infarction area of each implanted group obviously decreased (P < 0.05). Of them, the decrease was most obvious in the 4:1 group (11.37% +/- 2.18%) and the 8:1 group (9.23% +/- 2.35%, P < 0.05). Compared with the model group (cell/high magnification, 5.17 +/- 2.31), the Ki-67 positive cell number of each implanted groups significantly increased (P < 0.05). Of them, the Ki-67 positive cell number was obviously higher in the 8:1 group (15.00 +/- 3.16, P < 0.05).

CONCLUSIONSEPCs pretreated by salvianolic acid B combined with BMSCs could reduce the myocardial infarcted area, improve the proliferation of BMSCs in the peripheral infarction and local ischemia. Besides, along with the increase of the implant proportion of EPCs, the infarct area was gradually reduced, and the proliferative expression was gradually enhanced.

Animals ; Benzofurans ; pharmacology ; Bone Marrow Cells ; cytology ; drug effects ; Cell Proliferation ; Endothelial Cells ; cytology ; drug effects ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Myocardial Infarction ; metabolism ; pathology ; Rats ; Rats, Wistar ; Transplantation Conditioning

OBJECTIVETo investigate the role of AT2 receptors in the secretion of tumor necrosis factor alpha (alpha-TNF) and interleukin 1 beta (IL1 beta) in adult rat cardiac fibroblasts.

METHODSAdult rat cardiac fibroblasts in in vitro culture were divided into control, Ang II, AngII + Losartan, and AngII + PD123319 groups with corresponding treatments. Radioimmunoassay was used to determine alpha-TNF and IL1 beta levels in the supernatant of the treated cardiac fibroblasts.

RESULTSAng II treatment resulted in significantly increased alpha-TNF and IL1 beta levels. Compared with AngII group, IL1 beta level was decreased by 69.1% and 78.7% and alpha-TNF by 58.7% and 65.9% after blocking AT1 and AT2 receptors, respectively.

CONCLUSIONAT2 receptors are involved in alpha-TNF and IL1 beta secretions in cardiac fibroblasts.

Angiotensin II ; pharmacology ; Animals ; Fibroblasts ; drug effects ; metabolism ; secretion ; Gene Expression Regulation ; drug effects ; Interleukin-1beta ; secretion ; Male ; Myocardium ; cytology ; Rats ; Rats, Sprague-Dawley ; Receptor, Angiotensin, Type 2 ; metabolism ; Tumor Necrosis Factor-alpha ; secretion

AIM To investigate the effect of Shenfu Injection on atherosclerosis (AS) models of high-fat apolipoprotein E-deficient (ApoE-/-) mice,and to explore its anti-atherosclerosis mechanism through the observation of oxidative stress (OS) variation.METHODS C57 mice were used as controls.ApoE-/-mice fed with 20-week high fat diet were randomly divided into model group,Shenfu group for subsequent 4-week continuous corresponding intervention,after which the mice had their blood lipid levels measured,their levels of MPO and NOX4 identified by ELISA,and their T-SOD activity determined by hydroxylamine method,their MDA level detected by TBA,their plaque formation observation achieved by HE staining of aortic gross and red O of all the aorta,and their Nrf2 mRNA expression detected by real time qPCR method.RESULTS Compared with the control group,the model group manifested with increased contents of TG,TC,LDL,decreased HLD;decreased activity of SOD,increased contents of MPO,NOX4 and MDA,and down-regulated expression of aortic Nrf2 and Keap1 mRNA.Compared with the model group,Shenfu Injection group was observed with no obvious blood lipid level change,but a reduction of plaque area,and an effective inhibition on OS as revealed by improved levels of Nrf2 and Keap1 mRNA.CONCLUSION Shenfu Injection can activate Nrf2 and interfer the relevant enzymes,thus prevents the atherosclerosis progression through OS reduction.

Objective To study the influence on the tumor after percutaneous intra-tumor injection of ~(32)P-GMS in liver cancer as well as its suitable dose.Methods 24 New Zealand rabbits were used to establish the animal model of VX-2 liver cancer,and divided into A,B,C and D groups with individually 37,74,111 and 148 MBq of ~(32)P-GMS being injected,respectively;and then pathological changes of tumor were observed by light and electron microscope respectively.Result The dose of ~(32)P-GMS was obviously correlated with the radioactivity damage of tumor cells.In the A and B groups,the tumor cells were not observed to disappear completely after injection of ~(32)P-GMS,but in C group,tumor cells were almost completely disappeared and surrounded by a lot of connective tissue.Although the tumor cells were found to disappear completely in D group,normal liver tissues were also involved.Conclusion Percutaneous intra-tumor injection of ~(32)P-GMS with suitable dose that may induce the tumor tissue to be maximally damaged and may also provide some significances to prevent the tumor metastasis.

This study was aimed to investigate the expression of plasma miR-223 in pediatric acute lymphoblastic leukemia (ALL) in different treatment time point. A total of 64 pediatric ALL samples were selected from patients treated in Beijing Children's Hospital from May 2005 to January 2012, including 30 samples at new diagnosis (ND), 30 samples at complete remission (CR) and 4 samples at relapse. Without RNA extraction, the miR-223 levels in plasma were directly detected by a reverse-transcription quantitative real-time PCR assay. The results indicated that the expression of plasma miR-223 in pediatric ALL was lower at ND but elevated after CR. The miR-223 expression in plasma of relapse patients didn't show significant difference probably due to a few cases of relapse. The miR-223 levels in plasma had not displayed significant difference between TEL-AML1 positive patients and no fusion gene B lineage ALL patients either at ND or at CR. It is concluded that the plasma miR-223 decreases at ND and increases in CR of children with ALL. miR-223 may act as an anti-oncogene and may be taken as a potential predictive biomarker for evaluating the therapeutic effect of leukemia.
Child ; Child, Preschool ; Female ; Humans ; Male ; MicroRNAs ; blood ; genetics ; Plasma ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods

OBJECTIVETo establish a diagnostic model of protein fingerprint pattern in the cerebrospinal fluid (CSF) for non-small-cell lung cancer (NSCLC) patients with brain metastases.

METHODSThe CSF samples were obtained from 29 NSCLC patients with brain metastasis, 23 non-tumor patients and 10 early-stage NSCLC patients without brain metastases for analysis of the protein expression profiles using surface-enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS). The data were then analyzed by Biomarker Wizard software, and the tree analysis patterns were generated using the decision-tree model in Biomarker Patterns software. The diagnostic model was tested for its clinical application.

RESULTSFive protein peaks were identified showing differential expression between patients with brain metastases and those without brain metastases. Combination of the 3 protein peaks (m/z: 8698.00, 1215.32 and 1245.70) could discriminate these two samples with a sensitivity of 100.00% (29/29) and a specificity of 100.00% (23/23). Five proteins were differentially expressed between the NSCLC patients with brain metastases and the non-tumor patients. With one protein peak (m/z: 6050.00), these two samples could be discriminated with a sensitivity of 90.00% (9/10) and a specificity of 78.26% (18/23).

CONCLUSIONThe established diagnostic model of CSF protein fingerprint pattern provides high sensitivity and specificity in the diagnosis of NSCLC with brain metastasis.

Adult ; Aged ; Brain Neoplasms ; cerebrospinal fluid ; diagnosis ; secondary ; Carcinoma, Non-Small-Cell Lung ; cerebrospinal fluid ; diagnosis ; pathology ; secondary ; Cerebrospinal Fluid Proteins ; genetics ; Decision Trees ; Early Detection of Cancer ; Female ; Gene Expression Profiling ; Humans ; Lung Neoplasms ; cerebrospinal fluid ; diagnosis ; pathology ; Male ; Middle Aged ; Peptide Mapping ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVERespiratory syncystial virus (RSV) is the most common cause of lower respiratory infections in infants worldwide. There is no reliable vaccine or antiviral drug against RSV at present. RNA interference (RNAi) technology is a potent method to degrade expression of the cognate mRNA. In order to inhibit the replication of RSV at gene level, the effects of specific RNAi against M2-1 gene of RSV on inhibition of viral replication in cell culture system was observed in this study.

METHODSRSV M2-1 gene, which plays a key role in RSV transcription, was chosen in this study and was used as target gene and recombinant plasmid pshRNA7816 targeting the mRNA of RSV M2-1 gene coding sequence was constructed. The pshRNA7816 was transfected into Hep2 cells. The effects of the pshRNA7816 on changes of cytopathogenic effect (CPE) of Hep2 cell induced by RSV infection were observed microscopically. Viral plaque forming assay and MTT assay were used to detect the viral titer change and protective function of the pshRNA7816 on RSV infected Hep2 cell.

RESULTSThe recombinant RNAi plasmid pshRNA7816 which targets the mRNA of RSV M2-1 gene was successfully constructed. The pshRNA7816 significantly reduced CPE of RSV infected Hep2 cells, reduced the viral titer of RSV in the cells (P < 0.001). The pshRNA7816 raised the survival rate of RSV infected Hep2 cells (P < 0.001). Non-specific pshRNA plasmid did not show anti-RSV effects (P > 0.05).

CONCLUSIONThe recombinant pshRNA7816 plasmid which targeted the mRNA of RSV M2-1 gene showed a significant and specific anti-RSV effect.

Hep G2 Cells ; Humans ; Plasmids ; biosynthesis ; RNA Interference ; RNA, Small Interfering ; biosynthesis ; RNA, Viral ; genetics ; Respiratory Syncytial Virus Vaccines ; biosynthesis ; Respiratory Syncytial Virus, Human ; drug effects ; physiology ; Viral Proteins ; genetics ; Virus Replication ; drug effects