Potential role and mechanism for high mobility group box1 in childhood chronic immune thrombocytopenia.

OBJECTIVE: High mobility group box1(HMGB1) can be used as PAMP or alarmins to stimulate the innate immune system; however, previous research on immune thrombocytopenic purpura (ITP) mainly focused on its adaptive immunity. The study aimed to determine whether HMGB1 is associated with chronic ITP (cITP) during childhood and investigate its role in innate immunity in childhood cITP. PATIENTS AND METHODS: We recruited 80 patients to measure the expression of HMGB1, IL-17, and IL-10; 55 patients were recruited to measure the expression of TLR2 and TLR4 in monocyte and CD1c+dendritic cells, and 30 volunteers were included as controls. We focused on the expression of the NLRP3 inflammasome during childhood cITP. Furthermore, the impact of HMGB1 on the NLRP3 inflammasome was explored. RESULTS: The expressions of HMGB1 and IL-17 increased in children with cITP, while that of IL-10 decreased; HMGB1 was correlated with the expression of IL-17 and IL-10. The expression of TLR2 in CD14++CD16+, CD14+CD16++ monocytes increased significantly in comparison with the controls; the contrary was observed regarding TLR4. The expression of NLRP3, IL-1ß, and IL-18 was significantly higher in CD14 and CD1c, respectively. As the concentration of HMGB1 increased, the expression of NLRP3, IL-1ß, and IL-18 increased in different degrees. CONCLUSIONS: HMGB1 could be used as an early warning alarm for childhood cITP and is involved in developing cITP. HMGB1 could affect the incidence and development of chronic childhood ITP via the NLRP3, TLR2/TLR4 pathways.

Relationship between brain activity and voiding patterns in healthy preterm neonates.

INTRODUCTION: It remains controversial as to whether the brain affects voiding control in preterm newborns. Constant bladder volume has previously been thought to induce bladder voiding in neonates, with no influence from the brain. Lately, there has been distinct evidence for an existing connection between the central nervous system and bladder voiding in preterm infants, as the voiding reflex arouses neonatal children. Video electroencephalography (EEG) is useful for recording bioelectrical activity of the cerebral cortex and exploring its relationship with voiding patterns in preterm neonates. OBJECTIVE: The objective was to investigate the relationship between voiding patterns and brain activity in healthy preterm neonates by using video-EEG. STUDY DESIGN: Forty-seven healthy preterm neonates (16 females) with a mean postconceptional age (PCA) of 34.1 ± 1.8 weeks were divided according to PCA into three groups: Group I (31-33 weeks, n = 13); Group II (33-35 weeks, n = 14); and Group III (35-37 weeks, n = 20). Video-EEG data from eight cortical regions were recorded from 08:00-12:00, along with 4-hour free voiding patterns and status at voiding (awake/sleep). RESULTS: In Group I, the voiding frequency (VF) was significantly higher and the voiding volume (VV) was significantly lower than in the other groups. There were no significant differences in bladder capacity (BC), bladder capacity/birth weight (BC/BW), postvoiding residual/bladder capacity (PVR/BC), or urinary flow rate (UFR) among the three groups. The Fp1-T3 and Fp2-T4 lead amplitudes significantly differed in Group I and Group II at 5 s before (pre-5), during, and after voiding (post-5). The Fp2-C4 total and theta band lead amplitudes significantly differed across all urination states among the groups. There were no significant differences in electroencephalography frequency among the groups in any urination state. DISCUSSION: There were no significant differences in BC, BC/BW, PVR/BC, or UFR among the three groups, indicating slow bladder function development in preterm neonates. In this study, the EEG amplitude changed in certain pairs of electrodes. These changes might indicate the degree of bladder sensor maturation along with an increasing PCA. This study further suggests that the brain changes in preterm neonates during quiet sleep voiding prominently occur in the right prefrontal cortex and central region. CONCLUSIONS: In preterm neonates, bladder voiding during quiet sleep was accompanied by cortical arousal that might have emanated from a lower center.

Rapid generation of Epstein-Barr virus-specific T cells for cellular therapy.

Lymphoproliferative disease after hematopoietic stem cell transplantation is a serious and life-threatening disease. CD8(+) T lymphocytes play a key role in the control of viral diseases and some tumors. Adoptive immunotherapy has been shown to be effective in controlling and preventing some virus-related diseases in transplant recipients. However, the ex vivo production of cells for adoptive transfer is labor-intensive and expensive. To simplify the culture procedures of Epstein-Barr virus (EBV)-specific cytotoxic T lymphocytes, we explored a novel method, a rapid expansion protocol, that only used recombinant human interleukin to initiate the growth of clinical-grade EBV-specific T cells. After restimulation with EBV peptides principally from latent antigens and immediate/early antigens, the CD8(+) cells obtained could produce cytokines that had cytotoxic activity against target cells. This method provides a new possibility for the treatment of life-threatening EBV-associated malignancies.

AP4M1 is abnormally expressed in oxygen-glucose deprived hippocampal neurons.

AP4M1 mutations have been suggested to be associated with autosomal recessive cerebral palsy syndrome. But the pathogenic mechanism remains uncertain. The purpose of this study is to investigate whether and how AP4M1 expression is changed in injured neurons. Primary cultured hippocampal neurons were prepared for this experiment. They were subjected to oxygen-glucose deprivation (OGD) leading to apoptosis, mimicking brain ischemia. Neuron-specific enolase (NSE) was labeled immunofluorescently to confirm that the purity of neuron was higher than 90%. Real-time PCR and western blotting were performed to measure the gene expression. AP4M1 was labeled with MAP2 or Tau-1 to observe the distribution. We found that the AP4M1 protein levels immediately after the procedure were similar between the OGD group and the sham group. However, down-regulation was observed 12h after the reperfusion, and became more notable at 24h. The real-time PCR showed similar results, except that the down-regulation of mRNA was able to be detected immediately after the OGD. Immunofluorescent labeling revealed AP4M1 distributed in the dendrites of normal neurons, but it redistributed to the axons after the OGD procedure. In conclusion, AP4M1 is not only down-regulated at both the mRNA and protein levels, but also redistributed from dendrites to axons in oxygen-glucose deprived hippocampal neurons.

Effect of small interfering RNA targeting wild-type FLT3 in acute myeloid leukaemia cells in vitro and in vivo.

This study investigated the effect of using small interfering RNA (siRNA) to silence the wild-type FMS-like tyrosine kinase 3 (FLT3) gene in acute myeloid leukaemia (AML) cells, in vitro and in vivo. FLT3 siRNA was introduced into the human AML cell line, THP1, and into a THP1 xenograft tumour model in BALB/c nude mice. FLT3 siRNA effectively reduced both the mRNA and the protein levels of FLT3, arrested cells in G(0)/G(1) phase, inhibited THP1 cell proliferation and increased apoptosis. Intraperitoneal injection of FLT3 siRNA suppressed tumour growth in BALB/c nude mice. FLT3 siRNA treatment also reduced cyclin D1 and Bcl-2 protein levels, and increased the nuclear level of silencing mediator for retinoic acid and thyroid hormone receptors protein both in vitro and in vivo. These data suggest that FLT3 siRNA is a strong inhibitor of FLT3 expression in vitro and in vivo, and may provide a new therapeutic target for AML.

Seasonal variation of the particle size distribution of n-alkanes and polycyclic aromatic hydrocarbons (PAHs) in urban aerosol of Guangzhou, China.

Seasonal aerosol samples have been collected by Andersen Hi-Vol pumping system equipped with a five stage cascade impactor and a backup filter (size range: 10-7.2 microm, 7.2-3.0 microm, 3.0-1.5 microm, 1.5-0.95 microm, 0.95-0.49 microm,

Development of a compound-specific carbon isotope analysis method for atmospheric formaldehyde via NaHSO3 and cysteamine derivatization.

A novel method has been developed for the compound-specific carbon isotope analysis of atmospheric formaldehyde using gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). The method allows the determination of the delta13C value for atmospheric formaldehyde at nanogram levels with higher precision and lower detection limit. In the present work, atmospheric formaldehyde was collected using NaHSO3-coated Sep-Pak silica gel cartridges, washed out by water, then derivatized by cysteamine of known delta13C value, and the delta13C value of its derivative (thiazolidine) determined by GC/C/IRMS. Finally, the delta13C value of atmospheric formaldehyde could be calculated by a simple mass balance equation between formaldehyde, cysteamine, and thiazolidine. Using three formaldehydes with different delta13C values, calibration experiments were carried out over large ranges of formaldehyde concentrations. The carbon isotope analysis method achieved excellent reproducibility and high accuracy. There was no carbon isotopic fractionation throughout the derivatization processes. The differences in the carbon isotopic compositions of thiazolidine between the measured and predicted values were always <0.5 per thousand, within the specifications of the GC/C/IRMS system. The present method was also compared with the previous 2,4-dinitrophenylhydrazine derivatization method, and this method could be performed with lower analytical error and detection limit. Using this method, four 6-h ambient atmospheric formaldehyde samples were consecutively collected from 8 to 9 March 2005. The results showed that the delta13C values of atmospheric formaldehyde were different during the daytime and nighttime. This method proved suitable for the routine operation and may provide additional insight on sources and sinks of atmospheric formaldehyde.

[The determination of porphyrin carbon isotope composition by gas chromatography-isotope ratio monitoring mass spectrometry technique].

The porphyrin carbon isotope composition can be used to explore the precursor of porphyrin, oil-oil and oil-source rock correction and calculation of paleo P CO2. The conventional method is limited because of its time consuming and large sample size (several mg of individual porphyrin) required. Therefore, it hampers the application of porphyrin carbon isotope composition into the chemistry and geoscience. The present paper describes a quantification method to prepare bis-(tert-butyldimethylsiloxy) silicon (IV) [(TBDMSO)2Si(IV)] porphyrin which is sufficiently volatile at 300 degrees C and can be used for GC-IRMS analysis. The analysis of carbon isotope composition of aetio I as the form of free base, nickel, demetalization derivative, silicon(IV) and (TBDMSO)2Si(IV) have shown that aetio I porphyrin has no obvious isotope fractionation in the whole synthesis procedure for (TBDMSO)2Si(IV) porphyrin. The carbon isotope study on the porphyrin mixtures of aetio I and OEP indicates that isotope exchange between porphyrins during the synthesis of (TBDMSO)2Si(IV) porphyrin is absent. The method can be applied to the determination of porphyrin carbon isotope compositions. The advantages of the method are time saving, less sample size and lower standard deviation.

[A study on the contents of fat and fatty acids in five shellfishes].

The contents of fat and fatty acids in Callista erycina (Linnaeus), Paphia (Paratapes) undulata (Born), Meretrix meretrix (Linnaeus), Chlamys farreri (Jones et Preston) and Patinopecten yessoensis (Jay) were studied. Fat was extracted with Bligh & Dyer (B&D) method. The lipid classes were transesterified with potassium hydroxide in methanol. Fatty acid methyl esters (FAMEs) were assayed with GC-MS and polar capillary column (HP-INNOWax 30 m x 0.25 mm i.d. x 0.25 micron). GC injector temperature was 220 degrees C. The column temperature was programmed from 150 degrees C (1 min) to 200 degrees C at 10 degrees C/min and then from 200 degrees C to 250 degrees C at 2 degrees C/min. FAMEs were identified by MS library, and part by their standards. Total identified fatty acids were over 99% for all samples. Fat contents of them were all over 1% by wet samples. And ratios between omega-3PUFA and omega-6PUFA were above 2 by and large. Patinopecten yessoensis (Jay) contains more fat and the valuable fatty acids, EPA and DHA. It is suitable to use it as the source of EPA and DHA.

Fractionation and composition of colloidal and suspended particulate materials in rivers.

The association of pollutants (nutrients, heavy metals and organic compounds) with colloidal and suspended particle matter (SPM) plays a dominant role in determining their transport, fate, biogeochemistry, bioavailability and toxicity in natural waters. A scheme for the fractionation and composition of colloidal and SPM from river waters has been tested. All four separation methods, i.e. sieving, continuous flow centrifugation, tangential flow filtration, sedimentation field-flow fractionation, were for the first time used to separate five size particulate fractions from river. Significant (gram) amounts of colloidal material (<1 microm) in three size ranges, nominally 1-0.2, 0.2-0.006 and 0.006-0.003 microm were obtained. The separation scheme was able to process large samples (100 l), within reasonable times (1 day) and the apparatus was portable. The aquatic colloid size was also characterized with high resolution by using sedimentation field-flow fractionation technique. The mass-based particle size distribution for the water sample showed a broad size distribution between 0.05 and 0.4 microm with the maximum around 0.14 microm. There was a systematic increase in the content of organic carbon (estimated by loss on ignition), Mg, Ca, Na, Cu and Zn with decreasing particle size, highlighting the importance of the colloidal (<1 microm) fraction. It was concluded that the colloidal Cu and Zn concentrations in rivers might be much higher than those reported before.

[Stability of fatty acids in stored oyster].

The contents of twenty-three kinds of fatty acids in the lyophilized oyster that stored for 0, 15, 30, 45, 60, 75, 90 days were studied. Oyster fat was extracted from the powder by means of SFE. The extraction was performed for 40 min at a pressure of 37 MPa and a temperature of 50 degrees C with supercritical carbon dioxide containing 8% (V/V) ethanol at a flow-rate of 2 mL/min as liquid carbon dioxide, and the recovery of oyster fat by extraction was over 99%. After being extracted, fat was esterified with potassium hydroxide and methanol. Methyl esters of fatty acid were separated and determined by 30 m x 0.25 mm i.d. x 0.25 micron HP-INNOWax capillary column and MS detector. The injector temperature was 220 degrees C. The column temperature was programmed from 150 degrees C (1 min) to 200 degrees C at 10 degrees C/min and then from 200 degrees C to 250 degrees C at 2 degrees C/min. Twenty-three peaks were identified with gas chromatography/mass spectrometry, and quantified with area normalization method. The variations of contents of them were shown. During storage, the contents of saturated fatty acids and mono-unsaturated fatty acids were getting higher, and those of polyunsaturated fatty acids were getting lower. The decrease of them was gradual, and there was no special period of stability. And the stability of fatty acids in oyster related to the degree of unsaturation of them. The higher the unsaturation the lower the stable it was. After being stored for 90 days, the content of EPA decreased from 16.94% to 5.43% and that of DHA from 9.25% to 2.86%.