Objective To explore the activation and apoptosis of peripheral blood lymphocytes(PBLs) in children with Henoch-Schonlein purpura(HSP) and the effects of triptoIide(TP) on them. Methods The changes of activation and apoptosis were observed on cultured PBLs in children with HSP and healthy controls ,and the effects of TP were compared respectively. Expression of CD3, CD25 and apoptosis rate of PBLs were assayed with flow cytometry. Results The percentage of CD3+ CD25+ cell was significantly higher (P

Objective To investigate the correlation between cytotoxic T lymphocyte associated antigen 4 (CTLA - 4 ) and primary nephrotic sysdrome(PNS) with the pathologic type of mesangial proliferative glomerulonephritis(MsPGN) of glucocorticoid(GC) resistance. Methods The polymerase chain reaction - restriction fragment length polymorphism( PCR - RFLP) analysis was used to investigate the genotype of position - 318 promoter of CTLA- 4 gene of 36 patient children and 30 healthy controls. Results The frequences of genotypes at position - 318 promoter of CTLA- 4 gene in patients were 38.9% for CC,61.1 % for TC and 0% for TT. The frequences of alleles at this position were 69.4% for C allele and 30.6% for T allele. The frequences of genotypes and alleles in children were not significantly different from those in controls. Conclusion CTLA-4 promoter ( -318)C/T dimorphism was not associated with PNS- MsPGN of GC resistance, which hinted that the polymorphism may be not inlolved in pathogenensis of PNS-MsPGN and the mechanism of GC resistance.

OBJECTIVETo assess the anti-tumor immune response to percutaneous cryoablation in patients with local prostate cancer.

METHODSWe treated 10 patients with local prostate cancer by percutaneous cryoablation, collected the blood samples before and 2 weeks after the treatment and isolated peripheral blood mononuclear cells (PBMCs). Protein lysates were made by biopsy from autologous prostate cancer or non-cancer tissues. The levels of serum TNF-alpha, IFN-gamma, IL4 and IL-10 were determined by enzyme-linked immunosorbent assay (ELISA) and the Th1/Th2 ratio was calculated by the IFN-gamma/IL-4 ratio. The number of IFN-gamma + T cells under the stimulation of different protein lysates was counted by enzyme link immunol spot (ELISPOT). And the cytolytic activity of cytotoxic T lymphocytes (CTL) was detected by LDH assay.

RESULTSCompared with pre-treatment, the levels of TNF-alpha and IFN-gamma, the Th1/ Th2 ratio and the number of IFN-gamma + T cells induced by tumor protein lysates in PBMCs were increased significantly after cryosurgery (P < 0.01), while the levels of IL4 and IL-10 decreased slightly, and the non-tumor protein lysates induced no obvious changes in the number of IFN-gamma T cells. The cytolytic activity of cytotoxic T lymphocytes against human prostate cancer cells LNCaP was markedly increased, but not that against renal cancer cells GRC-1. One case of recurrence was found during the 3-6 months follow-up.

CONCLUSIONPercutaneous cryoablation for prostate cancer could induce a tumor-specific immune response.

Aged ; Cryosurgery ; Humans ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-4 ; blood ; Male ; Middle Aged ; Prostatic Neoplasms ; immunology ; therapy ; T-Lymphocytes, Cytotoxic ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo reveal the relationship between the storage time of the bark of Magnolia officinalis and the content of phenols in it, and lay a theoretical foundation for the harvest, processing, management and storage.

METHODThe contents of magnolol and honokoiol in 15 bark samples, collected from the main producing areas in China, were determined in the time of freshly harvest and 3 and 10 years after respectively by HPLC method.

RESULTIt showed that within a certain period of time, bark storage was favorable to conversion and accumulation of phenols, that the content of magnolol tended to increase from year 0 to year 3, then followed by slight decrease with years on account of volatilization of phenols, but was still higher when the bark was stored for 10 years than that that when the bark was freshly harvested, and the content of honokoiol still tended to increase when the bark had been stored for 10 years.

CONCLUSIONThe phenols in bark of M. officinalis is quite stable and the bark can be stored for 10 years or longer.

Biphenyl Compounds ; analysis ; Drug Storage ; methods ; Drugs, Chinese Herbal ; chemistry ; Lignans ; analysis ; Magnolia ; chemistry ; Plant Bark ; chemistry ; Time Factors

BACKGROUNDThe hairpin cell-penetrating peptides (hCPPs) demonstrate an interesting characteristic of conditioned activation by molecules. We hypothesized that hCPPs have the potential to selectively deliver a paramagnetic gadolinium probe into the matrix metalloproteinase 2 (MMP-2) positive human ovary adenocarcinoma cell lines, SKOV-3.

METHODShCPPs were synthesized and labeled with 1,4,7,10-tetraazacyclododecane-N,N',N'',N''' tetraacetic acid gadolinium (III) (Gd-DOTA) and fluorescein isothiocyanate (FITC) by f-moc strategy using a standard solid phase peptide synthesis protocol. MMP-2 expression and activity were demonstrated by reverse transcriptase polymerase chain reaction (RT-PCR) and zymography. Internalization and location of hCPPs in SKOV-3 cells were observed by fluorescein imaging and flow cytometery. Selective delivery of Gd-DOTA in SKOV-3 cells was observed by magnetic resonance imaging (MRI) and transmission electron microscopy (TEM).

RESULTSThe uptake of hCPPs by SKOV-3 cells depended on the activity of MMP-2. T1WI signals of SKOV-3 cells treated with Gd-DOTA-hCPPs suggested the uptake of Gd-DOTA-hCPPs increased in a time- (r = 0.990, P < 0.01) and concentration-dependent manner (r = 0.964, P < 0.001), but was inhibited by a MMP-2 inhibitor. Electron-dense particles observed in the cytoplasm and nucleus by transmission electron microscopy proved the intracellular penetration of gadolinium.

CONCLUSIONShCPPs can be used as an effective vector for an MRI molecular probe to assess the activity of MMP-2.

Cell Line, Tumor ; Cell-Penetrating Peptides ; adverse effects ; chemical synthesis ; chemistry ; metabolism ; Flow Cytometry ; Heterocyclic Compounds ; adverse effects ; chemical synthesis ; chemistry ; metabolism ; Humans ; Magnetic Resonance Imaging ; Matrix Metalloproteinase 2 ; chemistry ; metabolism ; Microscopy, Electron, Transmission ; Organometallic Compounds ; adverse effects ; chemical synthesis ; chemistry ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

BACKGROUNDThe cellular plasma membrane represents a natural barrier to many exogenous molecules including magnetic resonance (MR) contrast agent. Cell penetrating peptide (CPP) is used to internalize proteins, peptides, and radionuclide. This study was undertaken to assess the value of a new intracellular MR contrast medium, CPP labeled diethylenetriamine pentaacetic acid gadolinium (Gd-DTPA) in molecular imaging in vitro.

METHODSFluorescein-5-isothiocyanate (FITC) and Gd-DTPA respectively labeled with CPP (FITC-CPP, Gd-DTPA-CPP) were synthesized by the solid-phase method. Human hepatic cancer cell line-HepG2 was respectively stained by FITC-CPP and FITC to observe the uptake and intracellular distribution. HepG2 was respectively incubated with 100 nmol/ml Gd-DTPA-CPP for 0, 10, 30, 60 minutes, and imaged by MR for studying the relationship between the incubation time and T(1)WI signal. The cytotoxicity to NIH3T3 fibroblasts cells was measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide reduction assay (MTT).

RESULTSThe molecular weights of CPP labeled imaging agents, which were determined by MALDI mass spectrometry (FITC-CPP MW = 2163.34, Gd-DTPA-CPP MW = 2285.99), were similar to the calculated molecular weights. Confocal microscopy suggested HepG2 translocated FITC-CPP in cytoplasm and nucleus independent with the incubation temperature. MR images showed HepG2 uptaken Gd-DTPA-CPP had a higher T(1) weighted imaging (T(1)WI) signal, and that the T(1)WI signal intensity was increasing in a time-dependent manner (r = 0.972, P = 0.001), while the signal intensity between the cells incubated by Gd-DTPA for 60 minutes and the controlled cells was not significantly different (P = 0.225). By MTT, all concentrations from 50 nmol/ml to 200 nmol/ml had no significant (F = 0.006, P = 1.000) effect on cell viability of mouse NIH3T3 fibroblasts, compared with the control group.

CONCLUSIONSThe newly constructed CPP based on polyarginine can translocate cells by carrying FITC and MR contrast agent Gd-DTPA, and the intracellular concentrations are readily detectable by MR imaging, suggesting a new way for MR molecular imaging.

Cell Line, Tumor ; Cell Membrane Permeability ; Contrast Media ; Fluorescein-5-isothiocyanate ; Gadolinium DTPA ; Humans ; Magnetic Resonance Imaging ; methods ; Peptides ; metabolism

OBJECTIVETo elucidate the roles of Toll-like receptor 3 (TLR3) on dendritic cells (DCs) in HBV infection.

METHODSPeripheral blood mononuclear cells (PBMCs) were isolated from 48 healthy volunteers (HV) and 50 chronically HBV-infected patients (CH). DCs were induced and proliferated in a culture medium with rhGM-CSF and rhIL-4. We stimulated DCs with poly I:C and then TLR3, HLA-DR, and CD86, and CD1a expressions were examined by flow cytometry at 0 h, 12 h, 24 h and 48 h. The mRNA expressions of TLR3 were quantified by real-time PCR.

RESULTSTLR3 expression on DCs before the poly I:C stimulation and afterwards on the 12 h, 24 h, and 48 h were 69.2%+/-20.4%, 76.0%+/-18.6%, 78.2%+/-19.5% and 85.5%+/-6.9% respectively in the CH group, and 70.8%+/-11.2%, 67.5%+/-20.9%, 86.3%+/-14.7%, 68.6%+/-16.9% in the HV group. The expressions of TLR3 were up-regulated significantly at 24 h and 48 h after stimulation with poly I:C in the HV group, and in the CH group they were not significantly increased at 24 h but obviously increased at 48 h. The mRNA expressions of TLR3 increased significantly at 12 h in the HV groups, and at 48 h in CH group. The rate of CD86 expressions increased after poly I:C stimulation, and the increased rates were 12.6%+/-9.8%, 23.8%+/-20.0%, 20.7%+/-14.3% in the CH group, and 31.0%+/-25.0%, 43.4%+/-24.7%, 44.6%+/-25.5% in the HV group at 12 h, 24 h and 48 h after poly I:C stimulation. There was a marked increase of the expression level of CD86 in the HV group. In contrast, the level was only slightly increased in the CH group (31.0% vs 12.6%). The differences between the two groups were significant at 24 h and 48 h. No significant differences were detected in HLA-DR and CD1a between the two groups.

CONCLUSIONSThe increase of expression level of TLR3 is slower in the CH group than that in the HV group. A marked increase of the expression level of CD86 is observed in the HV group. Our results suggest that abnormal expression of TLR3 and CD86 may relate to the persistence of HBV infection.

Adult ; B7-2 Antigen ; metabolism ; Dendritic Cells ; immunology ; metabolism ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; blood ; Humans ; Male ; Middle Aged ; Toll-Like Receptor 3 ; metabolism ; Young Adult

Objective To analyze the cause, prevention and treatment of complications related to endovascular embolotherapy for ruptured intracranial aneurysms. Methods Sixty-eight patients with ruptured intracranial aneurysms received early endovascular embolization. Intraoperatively, artery spasm was relieved by papaverine infusion through microcatheter or balloon dilatation; parent artery occlusion by coils was treated by anticoagulation or antiplatelet treatment; ruptured aneurysms were embolized using mixed heparin; arterial thrombosis after embolization was ameliorated by the micro-catheter infusion of r-tPA thrombolytic. Results In the 68 cases, 8 cases suffered from complications, accounting for 11.77%. Among them, 1 case of extensive spasm of middle cerebral artery developed cerebral infarction and mild hemiplegia; 2 cases of localized spasm were improved well without sequelae; 1 case in which partial coils entered middle cerebral artery achieved a satisfied therapeutic outcome without infarction; 1 case with cerebral infarction due to coil dropping and 1 case with aneurysm rupture during operation received secondary embolotherapy without adverse consequences; one week after operation, 1 case died from aneurysm re-rupture; 1 case had hemiplegia owing to massive cerebral infarction. Conclusions It will do much benefit to secure the success of the operation by displaying the location, shape and size of the aneurysms as well as the relationship with the parent artery. Moreover, the proper choice of coils and well mastering of operative skills can decrease the incidence of complications, and adequate and prompt treatment of intraoperative complications can improve the prognosis of the patients obviously.

Mesenchymal stem cells (MSC) have been widely used in tissue regeneration and treatment graft versus host disease (GVHD) and immune diseases due to their self-renewal, multi-differentiation and immunoregulatory potential. However, more and more scholars begin to put weight on the MSC -derived extracellular vesicles (MSC-EV) for its regulation of inflammation and immunity. MSC-EV can activate the relevant signal pathways and regulate the function and biological behaviors of cells via acting on target cells and mediating communication between cells. MSC-EV has important potential clinical applications for its powerful immunomodulatory and hematopoietic regulatory functions. It is considered as a potential therapeutic tool to treat autoimmune diseases and GVHD. This paper reviewed the immunomodulatory activity of MSC-EV as well as the research progress of MSC-EV in hematopoietic stem cell transplantation, and discussed its potential clinical applications in the future.
Cell Differentiation ; Extracellular Vesicles/transplantation* ; Graft vs Host Disease/metabolism* ; Hematopoietic Stem Cell Transplantation ; Humans ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stem Cells

Objective The effect of different flow shear stress gradient on the changes of arrangement and shape of endothelial cells was evaluated in order to investigate the effect of shear stress gradient on ECs morphology and function. Method A flow chamber system with gradient shear stress was established, in which the range of shear stress is from 15 dyn/cm2 to 6.6 dyn/cm2(1 dyn=10-5 N), and the shear stress gradient is 1.5 dyn/cm2 and 3 dyn/cm2 respectively. After ECs were subjected to the gradient shear stress for 6 hours, cell angle, cell width length ratio, as well as cell shape index of ECs under the different shear stress gradient were examined. Results The cell angles of ECs were straggling under both 1.5 dyn/cm2 and 3 dyn/cm2 shear stress gradient. The cell width length ratio and cell shape index of ECs were decreased under 1.5 dyn/cm2 shear stress gradient compared with that of 3 dyn/cm2 shear stress gradient. Conclusions The ECs show random orientation under the different shear stress gradient. The ECs are trending to stretch and elongate shape under smaller shear stress gradient, and to approach cycloid under larger shear stress gradient.