Arrhythmias, Cardiac ; physiopathology ; Child ; Electrocardiography ; Humans ; Male ; Tachycardia, Ventricular ; physiopathology

Abstract: Objective　To explore the antiviral effect of baricitinib in the SARS-CoV-2 infection and influence on cytokine levels. Methods　Calu-3 cells were infected with SARS-CoV-2 at MOI of 0.1, and the levels of inflammatory cytokines (IL-6, IL-8, TNF-α and IL-1β), interferon β (IFN-β) and interferon-stimulated gene, IFIT2 in the infected cells were analyzed by qRT-PCR methods. At the same time, Calu-3 cells were infected with SARS-CoV-2 (MOI=0.1) after being treated with baricitinib for 2 hours. Cells were collected at 0, 24, 36, and 48 hours, and analyzed for the mRNA of the above genes in the drug-treated and untreated groups. Results　The mRNA levels of IL-6, TNF-a, IL-1β, IFN-β and IFIT2 in Calu-3 infected by SARS-CoV-2 cells were increased significantly. These cytokines were increased by nearly 100-fold post-infection 48 h compared with the control (P<0.000 1), and continued to increase with the infection time (P<0.001 or P<0.000 1). The increase of IL-8 mRNA level was not as significant as IL-6, TNF-α, IL-8, IL-1β, but it also showed a 2-4 folds increase. Baricitinib does not affect the level of viral RNA in Calu-3 cells after SARS-CoV-2 infection (P>0.05). However, baricitinib can significantly inhibit the up-regulation of IL-6 and TNF-α levels induced by SARS-CoV-2 infection (5.25-fold and 3.90-fold down-regulation, respectively, P<0.01), and has little effect on the levels of IL-8 and IL-1β . In addition, the drug could also significantly down-regulate the increase in IFN-β and IFIT2 levels caused by viral infection (10.51-fold and 90.78-fold down-regulation, respectively, P<0.000 1). Conclusions　Baricitinib inhibits the release of inflammatory cytokines to some extent, but it drastically down-regulates the expression of interferons and interferon-stimulated genes (ISGs), and has limited antiviral effect on SARS-CoV-2. Considering that interferon signal pathways play important roles on viral infection, caution should be exercised when using baricitinib to treat COVID-19 patients.

Objective Data on the leukapheresis from 26 pediatric patients with hematologic or solid malignancies was retrospectively evaluated to screen predictive factors affecting the efficacy of peripheral blood stem cell(PBSC) collection from donors,as well as hematopoietic recovery in recipients.Methods We present our experience with 49 apheresis from 26 granulocyte-colory Stimulating factor mobilized donors and analyzed the correlations between the mobilization,the leukocyte count in the donor peripheral blood and the MNC and CD_(34)~+ cell yields in collecting products and the neutrophil and platelet recovery of recipients.Results The process of mobilization and apheresis were well tolerated by our pediatric donors.The median numbers for harvested MNCs and CD_(34)~+ cells were 4.5?10~8/kg and 1.9?10~6/kg of recipient body weight,respectively.Mobilizing dose positively affected the number of mononuclear ceus(MNC) but not CD_(34)~+ cells in the apheresis products.The CD_(34)~+ cell number in the apheresis product was influenced significantly by donor circulating MNC on the day of harvest and correlated with recipient′s engraftment after PBSC was reinfused.Conclusions The MNC yield was stable and met with the demand for autologous stem cell transplantation while the CD_(34)~+ cell number varies obviously from each donor.Since a rapid engraftment was associated with a high number of CD_(34)~+ cells collected,which was in turn predicted by the level of the pre-apheresis CD_(34)~+ cells in the peripheral blood of donors,it is necessary to monitor the donors′ CD_(34)~+ cell during mobilization to determine the optimal time for apheresis.J Appl Clin Pediatr,2006,21(3):148-150

Objective To assess the feasibility of ultrasmall superparamagnetic iron oxide (USPIO)-enhanced magnetic resonance imaging (MRI) for monitoring the phagocytic activity in the brain tissue of rats following focal cerebral ischemia-reperfusion (IR) injury. Methods Forty male SD rats were randomized into 5 groups, namely 2, 3, 5-triphenyltetrazolium chloride (TTC) staining group (n=4), sham-operated group (n=6), and 3 cerebral IR injury groups with reperfusion time of 24, 48, and 72 h (n=10). USPIO was intravenously injected after focal cerebral IR injury, and MRI was performed at 24, 48, and 72 h after the reperfusion. The rats were sacrificed at 24, 48 and 72 h, and frozen sections of the local brain tissues were prepared to observe the cell death with HE staining, iron particle distribution with Prussian blue staining and the activity of the macrophages by CD68 immunohistochemical staining and immunofluorescent labeling. Results The ischemic lesions were identified as hyperintense area on T2-weighted images (T2WI) after middle cerebral artery occlusion (MCAO). The accumulation of USPIO appeared as hyperintense areas on T1WI and hypointense area on T2WI. The maximum signal change was observed at 24 h on T1WI (1.60±0.28) and at 48 h on T2WI (0.92±0.17) (P<0.05), and at each of the time points, the enhancement was significantly greater on T1WI than on T2WI (P<0.05). No obvious signal changes were found in the control group. Prussian blue staining detected iron oxide particles in both the peripherals of the ischcmic region and the necrotic area. A similar distribution pattern of the macrophagcs or activated microglia was found by CD68 immunohistochemistry and immunofluorescent labeling. Conclusion USPIO-cnhanced MRI allows dynamic monitoring of the inflammatory reaction in the local brain tissues aftcr focal cerebral IR injury.

OBJECTIVETo establish a method for determining the content of alpha-lipoic acid in New Zealand rabbit plasma.

METHODSAlpha-lipoic acid in the plasma samples was purified by solid-phase extractor and analyzed on an HYPERSIL C18 column with isocratic mobile phase consisting of potassium dihydrogen phosphate-acetonitrile (50:50, v/v) at a flow rate of 1.0 ml/min and detection wavelength of 230 nm.

RESULTSThe standard curve was linear in the range of 5-100 microg/L (r=1) and the average recovery was 77.4%-82.1%. The relative standard deviations of intra-day and inter-day assay were within 1.5%-8.9%.

CONCLUSIONThe method is sensitive, accurate and simple for determining plasma alpha-lipoic acid levels in New Zealand rabbits.

Animals ; Chromatography, High Pressure Liquid ; methods ; Rabbits ; Thioctic Acid ; blood

A comprehensively comparison of the chemical profiles between sun-drying BJ (NBJ) and sulfur-fumigated BJ (SBJ) was conducted by HPLC analysis and the discrepant peaks were identified or tentatively assigned by HPLC-ESI-MSn. A total of 32 chemical components were used for qualitative comparison. Meanwhile, a quantitative comparison of BJwere conducted by HPLC analysis and determining seven compounds from 3 NBJ and 3 SBJ samples dramatic chemical changes were found. After sulfur fumigation, the contents of flavonoids glycosides and phenolic acids were remarkably reduced, but the contents of flavonoids aglycones were significantly increased. Multivariate statistics, including principle component analysis (PCA) and partial least squares discriminate analysis (PLS-DA) were used to investigate the potential damaging effect of sulfur-fumigating process. The PCA score plots showed six samples were clearly classified into the sun-drying and sulfur-fumigating groups. And according to VIP >1, the most important chemical markers were apigenin, luteolin and 3,5-dicaffeoylquninic acid which could be used to distinguish NBJ and SBJ samples. Combining the results of qualitative and quantitative analysis, it showed that the sulfur fumigation has a significant effect on BJ.
Chromatography, High Pressure Liquid ; Chrysanthemum ; chemistry ; Fumigation ; Least-Squares Analysis ; Principal Component Analysis ; Sulfur

To investigate the effects of wt-p53 gene on proliferation and differentiation of K562 cells and to explore the feasibility of wt-p53 in leukemia gene therapy, pC53-SN(3), containing wt-p53 cDNA, and temperature-sensitive p53 mutant pN53cG(Val135) which behaved like wt-p53 at 32.5 degrees C, were introduced into p53-null K562 cells respectively by lipofectin mediated DNA transfection. In the presence of G418, K-SN(3) and K-pN53cG clones expressing P53 protein were selected. The effects of exogenous wt-p53 gene on the proliferation and differentiation of K562 cells were studied by detection of cell growth curves, leukemic colony formation, cell cycle analysis and DNA fragmentation, TdT-mediated dUTP nick end labeling (TUNEL) and benzidine staining. The results showed: (1) The level of p53 mRNA in K-SN(3) cells was lower than that in K-pN53cG cells by RT-PCR. (2) K-SN(3) and K-pN53cG(32.5 degrees C) cells proliferated more slowly than the control K562 cells, and their colony formation was obviously suppressed. The cells in G(0)/G(1) phase increased, and the cells in S phase decreased. These features were more obvious in K-pN53cG(32.5 degrees C). (3) K-pN53cG(32.5 degrees C) showed the feature of apoptosis and K-SN(3) showed the characteristics of erythroid lineage differentiation. It was indicated that exogenous of wt-p53 was capable of inhibiting the proliferation of K562 cells and inducing apoptosis of the cells at higher p53 level and interestingly, inducing the cells differentiation on erythroid lineage at lower p53 level.

OBJECTIVETo assess the effect of RNA interference-mediated gene silencing of plasma membrane-related Ca(2+)-ATPase-1 (PMR1) gene on the insulin secretion in islet beta cells NIT-1 in vitro.

METHODSA small interfering RNA duplex (siPMR1) corresponding to the nucleotides 337-357 of mouse PMR1 cDNA was introduced into NIT-1 cells via liposomes. The gene silencing effect was assessed by RT-PCR, and the total insulin level in the transfected cells was measured by radioimmunoassay.

RESULTSTransfection with siPMR1 resulted in obviously reduced PMR1 expression and increased insulin secretion in NIT-1 cells.

CONCLUSIONThe synthesized siPMR1 can significantly silence the expression of PMR1 and promote the secretion of insulin in the islet cells in vitro, which shed light on further studies of RNAi-based therapy of diabetes.

Animals ; Calcium-Transporting ATPases ; deficiency ; genetics ; Cell Line ; Gene Expression Regulation ; Insulin ; secretion ; Insulin-Secreting Cells ; metabolism ; secretion ; Mice ; RNA Interference ; RNA, Messenger ; genetics ; metabolism

OBJECTIVETo explore the influence of Candida albicans (Ca) on the motility and ultrastructure of human spermatozoa and its possible mechanism.

METHODSSemen samples obtained from 10 healthy volunteers by masturbation were prepared by the swim-up technique and sperm density to 40 x 10(6)/ml. The samples were then inoculated at 37 degrees C with different concentrations of a uropathogenic strain of Ca isolated from an outpatient, with initial fungi/spermatozoa ratios varying among 1:1 (Group A), 1:10 (Group B), 1:100 (Group C), 1:1000(Group D), and 1:10,000 (Group E). And Group F containing Ham's F-10 only was found as the negative control. Motion parameters were analysed by computer-aided sperm analyzer (CASA) at 0 hour, 1 hour, 2 hours and 4 hours respectively. Modalities of spermatozoa and possible adherence and/or agglutination were observed under the light microscope. Finally, all the samples were studied by transmission electron microscopy.

RESULTSDistinct adhesion of spermatozoa to Ca and agglutination were noticed. In all the motion parameters, progressive motility was affected most and dependent upon incubation time and bacterial concentration. Progressive motility showed a significant difference between Group A and the control (P < 0.01). With the prolongation of incubation time, other parameters were showing more and more differences. Analysis by electron microscopy revealed multiple ultrastructural damages.

CONCLUSIONCa significantly inhibits human sperm motility and decreases sperm viability in vitro. Its mechanism is possibly related to Ca's adhesion to human spermatozoa and the impairment inflicted by Ca to sperm ultrastructure.

Candida albicans ; isolation & purification ; physiology ; Candidiasis, Vulvovaginal ; microbiology ; Female ; Humans ; In Vitro Techniques ; Male ; Sperm Motility ; Spermatozoa ; physiology ; ultrastructure

OBJECTIVETo investigate the expression changes of intrinsic cytokines TGF-beta(1) and TNF-alpha, telomerase activity and bcl-2 during ongoing apoptosis of HL-60 and K562 cells induced by p53.

METHODSpN53cG (Val135), a temperature sensitive p53 mutant, which behaved like wild type p53 (wt-p53) at 32.5 degrees C, were introduced into p53-null HL-60 and K562 cells respectively by lipofectin. In the presence of G418, HL-60-pN53cG and K562 pN53cG clones expressing p53 protein were selected. The ongoing expression of intrinsic cytokines (TGF-beta(1) and TNF-alpha), bcl-2 oncogene and hTERT mRNA during the apoptosis of HL-60 and K562 cells induced by p53 and the effects of exogenous p53 gene, TGF-beta(1) and TNF-alpha antisense PS-ODNS on the apoptosis of HL-60 and K562 cells and the expression of bcl-2 were studied by RT-PCR, quantitative RT-PCR, DNA fragmentation, TdT-mediated dUTP nick end labeling (TUNEL) and flow cytometery. The levels of secreted TGF-beta(1) and telomerase activity were detected by ELISA and PCR-ELISA, respectively.

RESULTS(1) The expressions of intrinsic TGF-beta(1) and TNF-alpha mRNA were up-regulated, while that of bcl-2 and hTERT down-regulated. The levels of TGF-beta(1) in the supernatant of HL-60 and K562 cells were increased, and the level of telomerase activity decreased. (2) Antisense PS-ODNS of TGF-beta(1) and TNF-alpha could obviously inhibit the p53 inducing cell apoptosis, and restore bcl-2 mRNA and protein to pre-treated level.

CONCLUSIONSExogenous p53 induces leukemia cell apoptosis via up-regulating the expression of intrinsic TGF-beta(1) and TNF-alpha and down-regulating the expression of hTERT and bcl-2.

Apoptosis ; drug effects ; genetics ; DNA, Antisense ; pharmacology ; DNA-Binding Proteins ; Gene Expression Regulation, Neoplastic ; HL-60 Cells ; Humans ; K562 Cells ; Leukemia ; genetics ; pathology ; Mutation ; Plasmids ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Telomerase ; genetics ; Transfection ; methods ; Transforming Growth Factor beta ; genetics ; Transforming Growth Factor beta1 ; Tumor Necrosis Factor-alpha ; genetics ; Tumor Suppressor Protein p53 ; genetics ; physiology