OBJECTIVETo explore the effects of down-regulated tryptase expression in mast cells on the synthesis and release of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) of vascular endothelial cells.

METHODSTryptase-siRNA (small-interfering RNA) vector was constructed to inhibit tryptase expression in P815 cells. The medium of P815 cells treated by the tryptase-siRNA (RNAi-P815 group) or pure vector (P815 group) was collected and used to culture bEnd.3 cells. The messenger RNAs (mRNAs) of IL-6 and TNF-alpha in bEnd.3 cells and their protein levels in the medium were measured by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively.

RESULTSIL-6 and TNF-alpha mRNAs in bEnd.3 cells cultured in RNAi-P815-conditioned medium decreased significantly compared to those in P815-conditioned medium. Consistently, IL-6 and TNF-alpha protein levels in the medium of bEnd.3 of RNAi-P815 group were lower than those of P815 group.

CONCLUSIONReduced tryptase expression significantly inhibited the synthesis and release of IL-6 and TNF-alpha in vascular endothelial cells. RNA interference targeting tryptase expression may be a new anti-inflammatory strategy for vascular diseases.

Animals ; Cell Line ; Culture Media, Conditioned ; Down-Regulation ; genetics ; Endothelial Cells ; enzymology ; secretion ; Gene Expression Regulation, Enzymologic ; Interleukin-6 ; genetics ; secretion ; Mice ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transgenes ; Tryptases ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; genetics ; secretion

BACKGROUNDRenal biopsy is necessary for diagnosing the pathological changes of primary nephrotic syndrome (NS). However, it is invasive, time-consuming and can not be performed frequent on the same patient. Thus, development of a non-invasive and rapid diagnostic method may improve clinical patient management.

METHODSProteomic tool magnetic bead-based matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MB-based MALDI TOF MS) was applied to serum to determine peptidome patterns that are characteristic of different pathological changes.

RESULTSSerum specimen from 114 patients with NS (62 were minimal change disease (MCD), 30 were membranous nephropathy (MN), and 22 were focal segmental glomerulosclerosis (FSGS)) and 60 normal individuals were analyzed using MB-based MALDI TOF MS. The peptidome pattern was generated by genetic algorithms using a training set of 31 MCD, 15 MN, 11 FSGS and 30 normal individuals and was validated by an independent testing set of the remaining samples. The serum peptidome pattern, based on a panel of 14 peaks, accurately recognized samples from MCD, MN, FSGS and healthy control with sensitivities of 93.5%, 86.7%, 63.6% and 90.0%, and specificities of 98.2%, 94.4%, 100% and 89.5%, respectively. Moreover, one peptide from peptidome pattern was identified by liquid chromatography tandem mass spectrometry (LC MS/MS) as fibrinogen A.

CONCLUSIONDetection of the serum peptidome pattern is a rapid, non-invasive, high-throughout, and reproducible method for identifying the pathological patterns of patients with nephrotic syndrome.

Adult ; Female ; Humans ; Male ; Middle Aged ; Nephrotic Syndrome ; blood ; Peptides ; blood ; Proteomics ; methods ; Reproducibility of Results ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods ; Young Adult

Objective To study the role of dendritic cells in the function of a recombinant protein TFPR 1 as an adjuvant .Methods Bone marrow cells were collected from four-to five-week-old male BALB/c mice under aseptic conditions, and cultured with complete RPMI 1640 containing rmGM-CSF and rmIL-4 for six days.TFPR1 was added on day 6, and cells were incubated for another 24 hours.LPS was used as positive control , while PBS as negative .The morphology of dendritic cells was observed under an optical microscope and laser confocal microscope , cell surface makers (CD40,CD80,CD86 and MHCⅡ)were detected with flow cytometry, and the cytokines in the supernatant were detected with ELISA.Results Compared with negative control ,dendritic cells incubated with TFPR1 for 24 hours were significantly different in morphology as was observed by optical and laser confocal microscopes , but were similar to positive control .Most of the dendritic cells treated with TFPR 1 showed less adherence and became round , whose podosomes became shorter , and even disappeared .Actin distribution changed from two poles of the cell to the membrane .CD40,CD80, CD86 and MHCⅡon the cell surface were up-regulated on stimulation by TFPR1,as was detected by FACS.These results showed that TFPR1 was capable of promoting dendritic cell maturation .ELISA showed dendritic cells treated with TFPR 1 secreted high levels of cytokines(IL-6, IL-8 and TNF-α).Conclusion TFPR1 is capable of promoting dendritic cell maturation , and activating cells to produce cytokines , indicating that dendritic cells can play an important role in the function of TFPR 1 as a novel ad-juvant .

OBJECTIVETo probe blood serum Ab-IgG characteristics of severe acute respiratory syndrome (SARS) patients in Guangzhou and investigate the related factors.

METHODSThe serum of such population diagnosed as SARS convalescent patients, non-SARS patients, family consanguineous contraction persons, wild animal and vegetable salesman and community common people was collected. The lab detective method of ELISA was adopted for these serum samples. And the epidemic investigations for the SARS patients were also carried out.

RESULTSOf these populations, the detective rate of Ab-IgG for the clinic diagnosed SARS patients, which was 53.7%; That for the wild animal salesman and community common people were 16.7% and 0.9%, respectively. Among the clinic diagnosed SARS patients, the positive antibody detective rate was 90.4% for those which had specific contact history or infectivity, which was higher than that for other population. Among the specific contact history or infectivity cases, the antibody positive rate for the young and the old was lower than that for the adult. Meanwhile the difference did not exist among other cases. The antibody positive rate was identical between the male and the female. And the antibody detective rate was decreased by the month.

CONCLUSIONAs a whole SARS-CoV Ab-IgG detective rate for the clinic diagnosed SARS patients was 53.7% only. The reasons for that mainly lie in the wrong clinic diagnosis besides these factors such as age, hormone use and reagent and so on. The combination of lab detection results and epidemic investigation was propitious to the diagnosis veracity. It was impossible for the sub-clinic infection of SARS-CoV virus. The importance in the virus transmitting course need to be further studied.

Adult ; Antibodies, Viral ; blood ; Child ; China ; epidemiology ; Female ; Humans ; Immunoglobulin G ; blood ; Male ; Middle Aged ; SARS Virus ; immunology ; Seroepidemiologic Studies ; Severe Acute Respiratory Syndrome ; epidemiology ; immunology

To characterize long-term nonprogressors (LTNPs) and viremia controllers (VCs), infected with HIV-1 through contaminated blood donation or transfusion between 1992 and 1996 in Henan, China. LTNPs and VCs were defined by CD4+T lymphocyte (CD4) count and viral load (VL). Of 29,294 patients infected with HIV-1 via contaminated blood donation or transfusion that had conducted for more than 20 years, 92 were LTNPs/VCs. There were 70 LTNPs (0.24%), 43 VCs (0.15%), and 48 LTNPs+VCs- (0.16%). VCs had a significantly lower CD4 nadir, compared to LTNPs and LTNPs+VCs-, and no significant differences for the highest VL and HIV-1 DNA. Cases P4 and P5 were LTNPs, while their VL reached approximately 4.3 log copies/mL. P6 was a VC, but with CD4 < 500 cells/μL constantly. Data from the LTNPs/VCs cohort provided valuable information, future research is needed.

Parkinson's disease (PD) is the second most common neurodegenerative disease, but none of the current treatments for PD can halt the progress of the disease due to the limited understanding of the pathogenesis. In PD development, the communication between the brain and the gastrointestinal system influenced by gut microbiota is known as microbiota-gut-brain axis. However, the explicit mechanisms of microbiota dysbiosis in PD development have not been well elucidated yet. FLZ, a novel squamosamide derivative, has been proved to be effective in many PD models and is undergoing the phase I clinical trial to treat PD in China. Moreover, our previous pharmacokinetic study revealed that gut microbiota could regulate the absorption of FLZ