Animals ; Embryonic Stem Cells ; transplantation ; Hematopoietic Stem Cell Mobilization ; Humans ; Myocardium ; cytology ; Stem Cell Transplantation

Objective To compare the effects of intra-sinus thrombolysis by using urokinase and heparin anticoagulation alone for cerebral venous sinus thrombosis (CVST).Methods Consecutive inpatients with CVST were enrolled retrospectively.Their demographic and clinical data were collected.The treatment mainly consists of intra-sinus thrombolysis by using urokinase and heparin anticoagulation alone.The outcomes were evaluated according to the modified Rankin scale (mRS) at 6 months.The mRS score ＜ 2 was defined as good outcome,and ≥2 was defined as poor outcome.Results A total of 36 patients were enrolled,including intra-sinus thrombolysis by using urokinase (n =18) and heparin anticoagulation alone (n =18).Twenty-one had good outcomes and 15 had poor outcomes.After treatment,the recanalization rate (94.4％ vs.66.7％ ;x2 =3.850,P=0.041) of the urokinase thrombolysis group and the good outcome rate at 6 months (72.2％ vs.44.4％ ; x2 =3.827,P =0.046) were significantly higher than those of the heparin anticoagulation group.The proportion of the patients receiving intra-sinus thrombolysis by using urokinase of the good outcome group was significantly higher than that of the heparin anticoagulation group (60.0％ vs.18.2 ％ ; x2 =5.360,P =0.021).Multivariate logistic regression analysis showed that the intrasinus thrombolysis by using urokinase was an independent protective factor for good outcomes in patients with CVST (odds ratio,1.085,95％ confidence interval 1.024-1.361; P=0.023),and the high coagulation state was its independent risk factor (odds ratio,0.185,95％ confidence interval 0.049-0.611;P=0.004).None of the patients occurred symptomatic cerebral hemorrhage.Conclusions Intra-sinus thrombolysis with urokinase may improve the outcomes for patients with CVST,and it is superior to the heparin anticoagulation alone.

AIM:To study the type, degree and axial distribution of low vision astigmatism in preschool children.METHODS:A group of 3-6 years old children were selected for astigmatism screening, and statistical analysis was performed on the detected 445 eyes of 308 people.RESULTS:With more than 0.50D astigmatism criteria, astigmatism examination of 308 people, accounting for 36.2%, of which 137 eyes astigmatism, astigmatism 171 monocular.The five types of astigmatism were compound hyperopia 40.7%, mixed 35.5%, compound myopia 8.5%, myopia 8.3%, simple hyperopia astigmatism degree 7.0%;69.0% were mild, 16.6% moderate, 14.4% severe.Astigmatism axial distribution was with the rule for 54.9%, against the rule 28.8%, oblique 16.6%.In binocular astigmatism eyes, axial symmetry was in 35.8%, asymmetry in 64.2%.CONCLUSION:The main type of astigmatism in preschool children are compound hyperopia and mixed astigmatism.Astigmatism degree is mainly mild.With the increase of age, the detection rate of moderate and high astigmatism increased.

In this study, a composite cell model for evaluation of idiosyncratic drug-induced liver injury (IDILI) was established in vitro from the perspective of immune inflammation. And this model was used to evaluate the risk of IDILI for 2,3,5,4'-tetrahydroxy-cis-stilbene-2-O-β-glucoside (Cis-SG) and 2,3,5,4'-tetrahydroxy-trans-stilbene-2-O-β-glucoside (Trans-SG). To determine the low, medium, and high dosage of Cis-SG and Trans-SG, CellTiter-Glo^® 3D Cell Viability Assay was used to detect the effects of Cis-SG and Trans-SG on cell viability of HepG2 cells in three dimensional (3D) culture, and MTT assay was used to detect the effects of Cis-SG and Trans-SG on cell viability of THP-1 derived macrophages. THP-1 derived macrophages were incubated by Cis-SG and Trans-SG directly or supernatants from HepG2 cells incubated with them. Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of interleukin-1β (IL-1β) in the supernatants of the THP-1 derived macrophages. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) were used to determine the expression of apoptosis-associated speck-like protein (ASC), Nod-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase-1 (caspase-1), and IL-1β in THP-1 derived macrophages. The results showed that there was no effect on the secretion of IL-1β in THP-1 derived macrophages incubated by Cis-SG and Trans-SG directly. However, the secretion of IL-1β, the protein and mRNA expression of ASC, NLRP3, caspase-1, and IL-1β significantly increased in THP-1 derived macrophages incubated by supernatants from HepG2 cells incubated with 1, 5, and 25 μmol·L^-1 Cis-SG or 25 μmol·L^-1 Trans-SG. In summary, the composite cell model for evaluation of IDILI established in vitro has been successfully applied in testing Cis-SG and Trans-SG. This composite cell model is helpful to evaluate and screen drugs with IDILI risk in vitro preliminarily, which provides methods for predicting and solving the idiosyncratic liver toxicity of drugs.

Objective To analyze the VP1 and VP4 genetic region of enterovirus 71 (EV71)isolated from severe cases and mild cases with hanD-foot-mouth disease (HFMD) in Shanghai in 2011.Methods Five EV71 strains isolated from severe cases and five EV71 strains from mild cases in 2011 were included.Reverse transcription-polymerase chain reaction (RT-PCR) method was used to amplify and sequence the VP1 and VP4 genes of EV71,and then the sequencing results were compared with those of A,B,C genotype reference EV71 strains from GenBank by nucleotide alignment,amino acid alignment and phylogenetic tree analyses.Results The homogeneity between EV71 strains from severe cases and mild cases was 96.0％-98.1％ and 93.7％-99.5％ for VP1 and VP4 nucleotide sequences,respectively.The VP1 nucleotide sequences of 5 strains isolated from severe cases and 5 strains from mild cases in Shanghai shared 86.9％-98.2％ and 87.4％ 98.5％ identity with genotype C,respectively,while the homogeneity of VP4 nucleotide sequence was 85.5％-100.0％and 84.5％-99.5％,respectively.In addition,compared with the Fuyang EV71 strains (representative of C4 subtype),the strains from mild and severe cases shared homogeneity of 97.0％-98.2％ and 97.9％-98.5％ for VP1 gene,respectively,96.1％-100.0％ and 97.1％-99.5％ for VP4 gene,respectively.Among 3 strains isolated from severe cases,mutations at the residue 282 in the VP1 protein (N→S) and residue 7 in the VP4 protein (T→A) were discovered simultaneously.Conclusions The 10 EV71 strains isolated from severe and mild cases in Shanghai belong to subgenogroup C4.Among 3 strains isolated from severe cases,mutations at the residue 282 in the VP1 protein (N→S) and residue 7 in the VP4 protein(T→A) are discovered simultaneously.

Objective To evaluate the survival ability of enterovirus 71 (EV71) on different surface and under different climate.Methods Each 1 × 105 tissue culture infective dose 50 (TCID50)EV71 was added on different aseptic surface of plastic,rubber,cloth and wood,respectively.Then these materials were put into biotron (artificial climatic chamber) which could simulate different temperature and moisture.The viruses were recovered after a definite time and then inoculated into Vero cell.The cytopathic effect (CPE) was observed everyday to survey the survival ability of EV71 on different medium surface.Results The recovery rates of EV71 on medium surface ranged from 89 ％-93 ％.The survival time of EV71 on medium surface varied under different climatic conditions.The longest survival time of the virus was observed under the condition of 20 ℃ as the temperature and 80％ as the humidity.After 24 hours of incubation,the infectious titer on plastic surface reduced about 4 lg.After 72 hours of incubation,the infectious titer reduced at least 3.89 lg on cloth and wood surface.Conclusions Temperature and humidity can affect the survival time of EV71 on medium surface,which is longer in the condition of low temperature and high humidity.The survival ability of EV71 on natural cloth and wood surface is better than that on synthetic plastic surface.

OBJECTIVETo explore the influence of 1,4-benzoquinone (1,4-BQ) on proliferation of human bone marrow haematopoietic stem cells (hBM-HSCs) and human bone marrow mesenchymal stem cells (hBM-MSCs).

METHODSThe bone marrow samples were collected from a healthy donor. Methylcellulose semi-solid culture medium was used to culture the mononuclear cells of bone marrow in different culture systems. Colony-forming unit (CFU) assay was utilized to evaluate the proliferation of hBM-HSCs exposed to 1,4-BQ at the doses of 10, 25, 50 and 100 µmol/L and to observe the influence of 1,4-BQ on the Colony-forming unit-erythroid (CFU-E)/Burst-forming unit-erythroid (BFU-E), Colony-forming unit-granulocyte, macrophage (CFU-GM), Colony-forming unit-granulocyte, erythroid, macrophage, megakaryocyte (CFU-GEMM) in hBM-MSCs. MTT assay was used to detect the proliferation of hBM-MSCs exposed to 1,4-BQ at the doses of 1, 5, 10, 25, 50, 100, 200, 500 and 1000 µmol/L for 24 h, respectively, after hBM-MSCs were isolated, cultured and expanded.

RESULTSThe results of CFU assay indicated that numbers of CFU-E/BFU-E, CFU-GM and CFU-GEMM in 25, 50 and 100 µmol/L groups significantly decreased, as compared with control group (P < 0.05). However, no significant difference was found between the 10 µmol/L group and the control group. The results of MTT assay showed that the cellular viability of hBM-MSCs exposed to 1,4-BQ at the doses of 50 ∼ 200 µmol/L for 24 h significantly decreased in a dose-depended manner. When the exposure dose was higher than 200 µmol/L, the cellular viability of hBM-MSCs was lower than 5% which was significantly lower than that of control group (P < 0.05). When the exposure dose was lower than 25 µmol/L, there was no significant difference of cellular viability between exposure group and control group (P > 0.05).

CONCLUSIONThe results of the present study demonstrated that 1,4-BQ could inhibit the colony forming of hBM-HSCs and the relative viability of hBM-MSCs in vitro. The hematotoxicity induced by 1,4-BQ may be related to inhibiting the proliferation capacity of hBM-HSCs.

Benzoquinones ; toxicity ; Bone Marrow Cells ; cytology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Erythroid Precursor Cells ; Granulocyte-Macrophage Progenitor Cells ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology

Objective Using the mercury gravity manometer as reference, the accuracy of automated sphygmomanometers in blood pressure (BP) measurement in maintenance hemodialysis (MHD) patients was evaluated. Methods 86 MHD patients were selected. Brachial artery BP was measured by Omron HEM-907 portable automatic electric monitor and mercury gravity manometer simultaneously at the ann without fistula. BP measurements were repeated for 3～4 times for each patient during dialysis. The two BP measurement methods were compared according to ANSI/AAMI SP10:2002' Manual, electronic, or automated sphygmomanometers', which stated that the mean difference between the tested method and the reference method should not be larger than ± 5 mmHg, and the standard deviation (SD) of the difference should not be larger than ± 8 mmHg. Results The mean difference of systolic blood pressure between Omron HEM-907 portable monitor and mercury gravity manometer was - 1.6 mmHg with a SD of 5.48 mmHg. The mean difference of diastolic blood pressure between the two methods was - 2.68 mmHg with a SD of 5.02 mmHg. Conclusions Omron HEM-907 portable monitor was reliable for BP measurements in MHD patients, and could replace mercury gravity manometer in BP monitoring during dialysis session. The introduction of automatic BP monitoring during dialysis session will greatly reduce nurses' labor load.

In order to confirm the role that the 49th amino acid residue plays in enzymatic inactivity of Glutamine 49 phospholipase A2(Gln49-PLA2),site-directed mutagenesis of its 49th amino acid gene codon was conducted using PCR.Aspartic acid 49 phospholipase A2(Asp49-PLA2-Q49D-PLA2),the mutant of Gln49-PLA2 was expressed in E.coli with pET32a+ vector.The fusion protein,expressed as inclusion body,after being denatured,was on-column refolded and purified by immobilized metal affinity chromatography(IMAC),and then cleaved by Factor Xa.The mature Q49D-PLA2 mutant was obtained by Hitrap SP cation exchange and Superdex 75 gel filtration chromatography,with the recovery rate of 1.3%,and the specific activity of the mature Q49D-PLA2 mutant was 72 U/mg.It has been demonstrated that the 49th glutamine amino acid residue is the main reason in enzymatic inactivity of Gln49-PLA2 and the results are helpful for denatured protein refolding,especially in rich disulfide bonds conditions.

5.0mg/L were randomly divided into LMWH treatment group and low molecular dextran treatment group with 20 patients in each group.The patients in LMWH group were treated with 0.3ml LMWH subcutaneous injection in abdominal wall in every 12h for 1-4 d.The patients in low molecular dextran group were treated with 500ml low molecular dextran plus 20ml danshen root,intervenous drop infusion for 1-7d.Results The D-Dimer blood serum level in the gestational late period was significantly higher than that of nongravida group(P