Objective To investigate human enterovirus 71 (EV71) resistance to type Ⅰ interferon induced antiviral effect.Methods After type Ⅰ interferons (α,β ) were incubated with HeLa cells,recombinant type Ⅰ herpes simple virus ( HSV-1 ) with green fluorescent protein (GFP) was inoculated onto the HeLa cells.HSV-1 proliferation was observed by GFP expression and PCR.After EV71 was inoculated onto HeLa cells incubated with the same quantity of interferon,proliferation of EV71 were detected by RTPCR of 2A gene. Results Recombinant HSV-1 GFP expression and viral DNA replication obviously decreased in HeLa cells incubated with type Ⅰ interferon (α,β).However,EV71 effectively proliferated in the interferon irritated HeLa cell by RT-PCR.Conclusion HeLa cell irritated by type Ⅰ interferon (α,β) produced antiviral substance that inhibits HSV-1 proliferation.EV71 resisted the antiviral substance induced by type Ⅰ interferon and could significantly replicate in the HeLa cells.

In order to investigate the influence of cytokine combinations on proliferation and differentiation of human umbilical cord blood CD34(+) cells into megakaryocytes/platelets in vitro, the CD34(+) cells from human umbilical cord blood were amplified in serum-free medium StemSpan(SFEM) supplemented with several cytokine combinations by three-phase culture system. The effects of the cytokine combinations were compared. The results showed that at day 14 of the first culture phase, the CD34(+) cells cultured with cytokine combinations SCF + TPO + FL + IL-3 were amplified (11 000 ± 1 000) times, which were significantly higher than that of cells cultured with SCF + TPO + FL, but were not significantly different from that of cells cultured with SCF + TPO + IL-3 or SCF + TPO + FL + IL-3+ hydroxyl-corticosteroids. At day 7 of the second culture phase, the CD34(+) cells cultured with cytokine combination SCF + TPO + FL + IL-11 were amplified by (204666.7 ± 11718.9) times, which were significantly higher than that of cells cultured with SCF + TPO + FL + IL-3, but were not significantly different from that of cells cultured with SCF + TPO + FL + IL-11 + BMP4 + VEGF. At day 3 and day 6, the CD34(+) platelet-like cells accounted for about (39.8 ± 1.9)%, (39.7 ± 2.6)% and (25.5 ± 1.4)%, (23.1 ± 3.5)% cultured with SCF + TPO + FL + IL-11 and SCF + TPO + FL + IL-11 + BMP4 + VEGF, and significantly higher than that of the cells cultured with SCF + TPO + FL + IL-3. It is concluded that the cytokine combination of SCF + TPO + FL + IL-3 is most suitable cytokines combination for the amplification of CD34(+) hematopoietic progenitor cells. The cytokine combination of SCF + TPO + FL + IL-11 is preferred for the proliferation and differentiation of megakaryocytes, this study lays an experimental basis for investigating the proliferation and differentiation of CD34(+) into megakaryocytes/platelets in vitro.
Antigens, CD34 ; immunology ; Blood Platelets ; cytology ; Cell Differentiation ; Fetal Blood ; cytology ; immunology ; Humans ; Interleukin-11 ; pharmacology ; Interleukin-3 ; pharmacology ; Megakaryocytes ; cytology ; Stem Cell Factor ; pharmacology ; Thrombopoietin ; pharmacology

OBJECTIVETo investigate the toxicity of anti-CD25 monoclonal antibody (mAb) to rat cornea and its effects on the cytokines in the aqueous humour after penetrating keratoplasty (PKP), thereby evaluating the effect of anti-CD25 mAb in preventing corneal allograft rejection.

METHODSThe corneal toxicity of anti-CD25 mAb at 50, 100 and 200 microg administered via subconjunctival injection was evaluated in 12 SD rats by histological examination and transmission electron microscopy. Another 93 SD rats were randomized into 5 groups, and transplantation of corneal allograft from Wistar rats was performed in 4 groups with the other group as the normal control. The 4 allograft groups were treated with saline, 100 microg anti-CD25 mAb, 100 microg anti-CD25 mAb with 50 microg dexamethasone, and 50 microg dexamethasone, respectively. The graft rejection was observed, the aqueous humour levels of IFN-gamma and IL-4 were measured with ELISA, and IFN-gamma mRNA expressions in the grafts detected with RT-PCR.

RESULTSanti-CD25 mAb at 50 or 100 microg did not show significant toxicity on the cornea, but at 200 microg, the mAb caused swelling of the corneal stromal cells and endothelial cells. After corneal allograft transplantation, a significant delay in allograft rejection was observed in the 3 groups with mAb or dexamethasone treatment as compared with that in saline group (P<0.05). IFN-gamma mRNA expression in the allograft on days 11 after PKP and in the aqueous humour on days 6 and 11 was markedly increased in saline group compared with that in the 3 treatment groups (P<0.05). The mean IL-4 level in the aqueous humour was significantly higher in the mAb group than in saline group (P<0.05), but markedly lower in anti-CD25 mAb+dexamethasone and dexamethasone groups than in anti-CD25 mAb group (P<0.05).

CONCLUSIONSAnti-CD25 mAb at 20 and 100 microg does not obviously affect the rat corneas. Anti-CD25 mAb inhibits IFN-gamma expression and promotes IL-4 the expression to reduce corneal allograft rejection, whereas anti-CD25 mAb with low-dose dexamethasone inhibits both IFN-gamma and IL-4 expressions to more effectively promote the graft survival.

Animals ; Antibodies, Monoclonal ; pharmacology ; Aqueous Humor ; drug effects ; metabolism ; Cytokines ; metabolism ; Female ; Graft Rejection ; prevention & control ; Interferon-gamma ; metabolism ; Interleukin-2 Receptor alpha Subunit ; immunology ; Interleukin-4 ; metabolism ; Keratoplasty, Penetrating ; methods ; Male ; Peptide Fragments ; metabolism ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar

OBJECTIVETo observe the therapeutic effect of tranquilizing and allaying excitement needling method on insomnia of heart and spleen deficiency and the effect of brain blood flow.

METHODSSixty cases were randomly divided into a tranquilizing and allaying excitement needling method group (observation group) and an eight confluence points selected group (control group), 30 cases in each group. The observation group was treated by acupuncture at Sishencong (EX-HN 1), Shenmen (HT 7), and Sanyinjiao (SP 6) with tranquilizing and allaying excitement needling method. The control group was treated by acupuncture at Shenmai (BL 62) and Zhaohai (KI 6). Their therapeutic effects and changes of brain blood flow were observed.

RESULTSThe total effective rate was 93.3% (28/30) in the observation group which was better than 83.3% (25/30) in the control group (P < 0.05). After treatment, the peak velocity of systolic (Vp) and diastolic blood flow velocity (Vd) of middle cerebral artery, basilar artery and vertebral artery were increased in the both groups (P < 0.01, P < 0.05), with more obvious increase in the observation group (P < 0.01, P < 0.05), except the Vd of left vertebral artery.

CONCLUSIONBoth the tranquilizing and allaying excitement needling method and the eight confluence points selected needling method can improve the clinical signs and symptoms. Meanwhile, the therapeutic effect of acupuncture is related with improvement of brain blood flow. However, the tranquilizing and allaying excitement needling method has better therapeutic effect on insomnia of heart and spleen deficiency.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Brain ; blood supply ; Female ; Heart ; physiopathology ; Humans ; Male ; Middle Aged ; Sleep Initiation and Maintenance Disorders ; physiopathology ; therapy ; Spleen ; physiopathology ; Young Adult

Acquired Immunodeficiency Syndrome ; epidemiology ; virology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Young Adult

This study was aimed to investigate a beneficial approach for resolving the deficiency of blood source, preventing the infection resulting from blood transfusion and overcoming the knotty match of patients with rare blood group by using massive expansion of erythroid cells from cord blood CD34(+) cells in vitro. The CD34(+) cells from human cord blood were cultured in serum-free medium supplemented with stem cell factor (SCF), interleukin-3 (IL-3) and erythropoietin (EPO) for 1 week, then expansion and differentiation of CD34(+) cells into erythroid cells were supported by co-culture with human mesenchymal stem cells (MSCs) derived from bone marrow for 2 weeks. The results indicated that after culture for 23 days, the expansion multiple of total cell number reached 2.52 x 10(5), and over 95% of these cells were erythroid cells as compared with less than 1% of myelomonocytic (CD14(+) or CD15(+)) cells and megakaryocytic (CD41(+)) cells. However, the culture system without MSC support was significantly disadvantaged both in expansion ability and ratio of erythroid cells when compared with MSC supporting system. It is concluded that the erythroid cells can be produced from CD34(+) cells in large scale by culturing in the system comprised of cytokine sets and MSC feeders, in which MSCs can support the proliferation and differentiation of erythroid cells.
Antigens, CD34 ; Cell Culture Techniques ; methods ; Cell Differentiation ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans

OBJECTIVETo analyse the molecular types of Listeria (L.) monocytogenes, and to construct the standard China L. monocytogenes pulsed-field gel electrophoresis (PFGE) subtyping database, using the international standardized L. monocytogenes-PFGE protocol.

METHODS118 L. monocytogenes strains isolated from 8 provinces or municipalities of China were subtyped according to L. monocytogenes-PFGE protocol.

RESULTS118 strains of L. monocytogenes were divided into 39 subtypes. In the 39 subtypes, 37 strains (31.36%) were GX6A160004 pattern, showing it was the predominant Lm subtype in China.

CONCLUSIONData from molecular typing suggested that the predominant Lm strains were distributed in different regions of China. PulseNet China-Lm database was constructed which was valuable for the molecular subtyping-based surveillance of L. monocytogenes.

China ; Electrophoresis, Gel, Pulsed-Field ; Food Microbiology ; Listeria monocytogenes ; classification ; genetics ; isolation & purification ; Phylogeny

Animals ; Bronchoalveolar Lavage Fluid ; Cigarette Smoking ; Interleukin-17/genetics* ; Lung ; MAP Kinase Signaling System ; Mice ; Mice, Inbred C57BL ; Pulmonary Disease, Chronic Obstructive

The polymorphisms of thyroid stimulating hormone receptor (TSHR) intron 1 rs179247 and rs12101255 have been found to be associated with Graves' disease (GD) in genetic studies. In the present study, we conducted a meta-analysis to examine this association. Two reviewers systematically searched eligible studies in PubMed, Web of Science, Embase and China Biomedical Literature Database (CBM). A meta-analysis on the association between GD and TSHR intron 1 rs179247 or rs12101255 was performed. The odd ratios (OR) were estimated with 95% confidence interval (CI). Meta package in R was used for the analyses. Seven articles (13 studies) published between 2009 and 2014, involving 5754 GD patients and 5768 controls, were analyzed. The polymorphism of rs179247 was found to be associated with an increased GD risk in the allele analysis (A vs. G: OR=1.40, 95% CI=1.33-1.48) and all genetic models (AA vs. GG: OR=1.94, 95% CI=1.73-2.19; AA+AG vs. GG: OR=1.57, 95% CI=1.41-1.74; AA vs. AG+GG: OR=1.54, 95% CI=1.43-1.66). The site rs12101255 also conferred a risk of GD in the allele analysis (T vs. C: OR=1.50, 95% CI=1.40-1.60) and all genetic models (TT vs. CC: OR=2.22, 95% CI=1.92-2.57; TT+TC vs. CC: OR=1.66, 95% CI=1.50-1.83; TT vs. TC+CC: OR=1.74, 95% CI=1.53-1.98). Analysis of the relationship between rs179247 and Graves' ophthalmopathy (GO) showed no statistically significant correlation (A vs. G: OR=1.02, 95% CI=0.97-1.07). Publication bias was not significant. In conclusion, GD is associated with polymorphisms of TSHR intron 1 rs179247 and rs12101255. There is no association between rs179247 SNPs and GO.
China ; Female ; Genetic Association Studies ; Genetic Predisposition to Disease ; Graves Disease ; genetics ; pathology ; Graves Ophthalmopathy ; genetics ; pathology ; Humans ; Introns ; genetics ; Male ; Polymorphism, Single Nucleotide ; Receptors, Thyrotropin ; genetics ; Risk Factors

ZNF804A rs1344706 has been identified as one of the risk genes for schizophrenia. However, the neural mechanisms underlying this association are unknown. Given that ZNF804A upregulates the expression of COMT, we hypothesized that ZNF804A may influence brain activity by interacting with COMT. Here, we genotyped ZNF804A rs1344706 and COMT rs4680 in 218 healthy Chinese participants. Amplitudes of low-frequency fluctuations (ALFFs) were applied to analyze the main and interaction effects of ZNF804A rs1344706 and COMT rs4680. The ALFFs of the bilateral dorsolateral prefrontal cortex showed a significant ZNF804A rs1344706 × COMT rs4680 interaction, manifesting as a U-shaped modulation, presumably by dopamine signaling. Significant main effects were also found. These findings suggest that ZNF804A affects the resting-state functional activation by interacting with COMT, and may improve our understanding of the neurobiological effects of ZNF804A and its association with schizophrenia.