Objective To investigate the frequency of c-kit mutation and prognosis in t (8;21) acute myeloid leukemia (AML) patients with trisomy 4. Methods A total of 145 de novo t(8;21) AML patients from February 2005 to January 2013 were analyzed retrospectively. Detection of exons 8 and 17 mutation of c-kit by PCR and cytogenetic analysis by R-banding technologies were performed on bone marrow samples of all patients at diagnosis. Clinical data were collected and analyzed statistically. Results Among 145 t (8;21) AML patients, 12 cases (8.3 %) were trisomy 4, 91.7 % (11/12) of them were identified with c-kit mutation, which was significantly higher than that without trisomy 4 [26.3 % (35/133), P< 0.01]. The follow-up data showed that the patients with trisomy 4 were correlated with the lower overall survival (OS) rate (15 % vs 56 %, P< 0.01) and disease-free survival (DFS) rate (0 vs 51 %, P< 0.01) when compared with patients without trisomy 4. Furthermore, the subgroup of patients with both trisomy 4 and c-kit mutation had a worse OS and DFS (P< 0.05). Conclusions Trisomy 4 is associated with high frequency of c-kit mutation and demonstrates poor prognosis in t(8;21) AML patients. Trisomy 4 or it combined with c-kit gene mutation is the main influencing factor on the survival of the patients with t(8;21) AML.

Objective To explore the operative methods and clinical outcomes in emergency or sube- mergency repair for the complicated tissue defects in hand in first stage applying microsurgical technique. Methods From Jan.,2000 to Aug.,2005,49 emergency cases of complicated hand tissue defects were re- paired in the first stage with replantation,reconstruction,free flaps,combined finger reconstruction and flap transplantation,including 21 cases in mini tissue mass replantation or reconstruction,15 cases in replantation combined with free flap transplantation,8 cases in replantation and reconstruction combined with free flap transplantation,5 cases in combined multiple digits reconstruction with free flap transplantation.The free flap transplantation included the anterolateral femoral flap,the latissimus dorsi myocutaneous flap,the dorsalis pe- dis flap,the media pedis flap and the instep flap.Results All the flaps,the replanted and reconstruceted finger survived uneventfully except for one replanted finger necrosis.45 cases healed in the first stage and the other 4 cases healed in the second stage.During a follow-up from 6 months to 3 years postoperatively,a satis- factory appearance and function of the reconstructed hand was achieved.The excellent and good rate was 85.7% assessed with provisional functional assessment criterion for upper limbs issued by Chinese Society of Hand Surgery.Conclusion The emergency or subemergency repair for the complicated tissue defects in hand has the advantage of short-term treatment and desirable functional outcome.The emergency replantation and reconstruction combined with various flaps or tissue mass can be applied to repair tissue defect in hand in the first stage according to the position and area of the defect along with the technique level of the surgeon, having been proved to achieve desirable clinical outcomes.And the key points leading to a successful operation is the correct treatment for the raw surface of the defects,suitable choice for various flaps,logical design of combination pattern and prevention and timely treatment for vessel crisis.

Objective To investigate the effect of recombinant adenovirus mediatied human endostatin (rAD-GFP-ES)on rats with collagen typeⅡinduced arthritis(CIA),and explore the mechanism of inflamma- tion and cytokines inhibition on rats CIA.Methods The rAD-GFP-ES was amplified and purified.The model of rat CIA was induced by intradermal injection of typeⅡcollagen combined with complete Freund's adjuvant(CFA). On the second day after the injection,the therapeutic administration of rAD-GFP-ES(1?10~(11)pfu?kg~(-1)?week~(-1)?4 weeks)were performed to the rats.The mean arthritis index(AI)was scored every week since then.The relative concentrations of ES,IL-I?,TNF-?in sera collected at the fourth week were evaluated by western blotting. Results①The titer of the purified rAD-GFP-ES and rAD-GFP was 6.6?10~(12)pfu/ml and 4.8?10~(12)pfu/ml,re- spectively(A_(260nm)/A_(280nm)＞1.3).②The concentration of ES in sera of the group treated with rAD-GFP-ES was 2.4-lold higher compared to the normal group.③The mean arthritis index of the group treated with rAD-GFP- ES was much lower than that of the model group.The administration of rAD-GFP-ES could significantly de- creas the production of IL-1?and TNF-?in sera.Conclusions①The rAD-GFP-ES is efficiently expressed in vivo.②The rAD-GFP-ES has an inhibitory effect on the arthritis index of rat CIA.③IL-1?and TNF-?are involved in the pathogenesis of RA.The rAD-GFP-ES has an inhibitory effect on the expression of IL-1?and TNF-?in rat CIA.

Animals ; Carps ; genetics ; Cloning, Molecular ; Fish Proteins ; genetics ; Hydroxylation ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; Lakes

ObjectiveTo observe the influence of iodine on mRNA expression of iodide transporter (NIS),insulin-like growth factor Ⅰ (IGF- Ⅰ ) and transforming growth factor beta(TGF-β) in thyroid and mammary glands of lactating rats and to explore the role of NIS,IGF- Ⅰ and TGF-β mRNA in iodine uptake in thyroid and mammary glands of lactating rats.MethodsOne hundred and one Wistar rats(80 female and 21 male),weighting 8 - 100 g were selected.These female rats were randomly divided into five groups according to their body weight:control group(NI,normal feed,drank deionized water containing iodine 50 μg/L) ; low iodine group 1 and 2(LI-1,LI-2,low iodine feed,drank deionized water containing iodine 0 and 5 μg/L,respectively); high iodine group 1 and 2(HI-1,HI-2,normal feed,drank deionized water containing iodine 3000 and 10 000 μg/L,respectively),16 rats in each group.After feeding for 3 months,the female and male rates were mated 3:1.The female rats in each group were sacrificed at the fifth and tenth day after postpartum.Thyroid and mammary glands were taken.The mRNA levels of NIS,IGF- Ⅰ and TGF-β in thyroid and mammary glands of lactating rats were determined by real time quantitative PCR.ResultsThe fifth days after postartum,NIS,IGF- Ⅰ and TGF-β mRNA expression levels of thyroid and lactating mammary glands were different between groups,and the differences were statistically significant ( NIS:F =631.46,64.91,all P ＜ 0.01 ; IGF- Ⅰ:F =11.45,6.56,all P ＜ 0.01 ; TGF-β:F =291.83,304.53,all P ＜ 0.01).Compared with control group [NIS:0.0066 ± 0.0023, (0.1481 ± 0.0711 ) × 10-2; IGF- Ⅰ:0.0419 ± 0.0062,0.0542 ± 0.0044; TGF-β:0.1416 ± 0.0277,0.1670 ± 0.0499],regardless of thyroid or mammary gland,the NIS,IGF- Ⅰ and TGF-β mRNA expression of LI-1 [NIS:0.0447 ± 0.0110,(0.3030 ± 0.1831) × 10-2;IGF- Ⅰ:0.0662 ± 0.0078,0.0902 ± 0.008; IGF- Ⅰ:0.5514 ± 0.0508,0.6942 ± 0.0367],LI-2[NIS:0.0317 ±0.0081,(0.3017 ± 0.1601) × 10-2; IGF-I:0.0645 ± 0.0054,0.0894 ± 0.0093; TGF-β:0.5292 ± 0.0332,0.6704 ± 0.0277 ] was significantly increased (all P ＜ 0.01 ); the NIS mRNA expression of HI-1 [0.0043 ± 0.0011,(0.1233 ± 0.0954) × 10-2],HI-2[0.0037 ± 0.0017,(0.1058 ± 0.0854) × 10-2] was decreased(all P ＜ 0.05),while the expression of IGF-Ⅰ mRNA [0.0521 ± 0.0910,0.0715 ± 0.0026; 0.0516 ± 0.0078,0.0697 ± 0.0038] and TGF-β mRNA [0.2087 ± 0.0425,0.2361 ± 0.0425; 0.1971 ± 0.0237,0.2257 ± 0.0752 ] was increased (all P ＜ 0.05 ).The tenth days after postpartum,the mRNA expression levels of NIS,IGF- Ⅰ and TGF-β of thyroid and lactating mammary gland in rats were different between groups,and the differences were statistically significant (NIS:F =103.55,116.32,all P ＜ 0.01; IGF-Ⅰ:F =67.67,11.98,all P ＜ 0.01; TGF-β:F =74.30,381.30,all P ＜0.01 ).Compared with the control group[NIS:0.0069 ± 0.0011,(0.1337 ± 0.0599) × 10-2; IGF-Ⅰ:0.0390 ±0.0071,0.0534 ± 0.0056; TGF-β:0.1351 ± 0.0336,0.1534 ± 0.0320],the mRNA expression levels of NIS,IGF- Ⅰ and TGF-β of LI-1 [ NIS:0.0432 ± 0.0165,(0.2962 ± 0.0985 ) × 10-2; IGF- Ⅰ:0.0643 ± 0.0088,0.0873 ± 0.0055 ; TGF-β:0.5042 ± 0.0912,0.6408 ± 0.0420],LI-2[NIS:0.0287 ± 0.0111,(0.2873 ± 0.0862) × 10-2; IGF- Ⅰ:0.0621 ± 0.0094,0.0862 ± 0.0038; TGF-β:0.4893 ± 0.0504,0.6372 ± 0.0389] were significantly increased(all P ＜ 0.01 ); the NIS mRNA levels of HI-1 [ 0.0042 ± 0.0029,(0.1006 ± 0.0909) × 10-2],HI-2[0.0035 ± 0.0020,(0.0890 ± 0.0119) × 10-2] were decreased(all P＜ 0.05),while the expression of IGF-Ⅰ mRNA[0.0516 ± 0.0078,0.0668 ± 0.0071; 0.0508 ± 0.0089,0.0621 ± 0.0064] and TGF-β mRNA[0.2007 ± 0.0546,0.2175 ± 0.0370;0.1959 ± 0.0393,0.2097 ± 0.0425] were increased(all P ＜ 0.05 ).In thyroid and mammary glands,the comparisons of NIS,IGF,TGF-β mRNA expression of the fifth and tenth day after postartum,between each group were not statistically significant(all P ＜ 0.05).ConclusionsThere are regulatory mechanisms of thyroid and mammary glands of lactating rats in response to low or high iodine conditions.In low iodine,the expressions of NIS,IGF- Ⅰ and TGF-β mRNA in thyroid and mammary glands increase and iodide uptake ability is enhanced to meet the body needs.In high iodine,the expression of NIS mRNA decreases in thyroid and mammary glands.Due to the reduced ability of iodine uptake,iodine intake is reduced,thereby reducing the hazards of high iodine in filial rats.

OBJECTIVETo establish a capillary electrophoresis-mass spectrometry(CE-MS) method for the analysis of nineteen organonitrogen pesticides in Paeoniae Radix Alba.

METHODSCE-MS analysis was performed on a 70 cm X 50 μm fused-silica capillary. The optimal buffer was composed of 1 % formic acid and 15 % methanol(V/V, pH 2.2). The temperature of capillary was controlled at 25 degree. The separation voltage was +20 kV. The optimal MS parameters were as follows: ESI-MS analysis was performed in the positive mode; 90 % methanol containing 0.2 % formic acid with a flow rate of 8 μl·min(-1) was selected as the sheath liquid; the flow rate and temperature of drying gas were 6 L·min(-1) and 250 degree, respectively; The nebulizing gas pressure was set at 5 psig; The optimal values of fragmentor and ESI voltage were 100 V and 5 000 V, respectively.

RESULTSThe nineteen pesticides had good linearity over the testing ranges. The average recoveries were in the range of 80.1 %-108.4 % with RSDs less than 20 % (except ethoxyquin and spiroxamine, those of which were 29.2 % and 22.3 % at 0.01 mg.kg(-1) concentration level). The LODs of nineteen pesticides were 0.503 ≊10.1 μg.kg(-1).

CONCLUSIONThe method can be used effectively to analyze the nineteen organonitrogen pesticides residue in Paeoniae Radix Alba.

Electrophoresis, Capillary ; methods ; Mass Spectrometry ; methods ; Paeonia ; chemistry ; Pesticide Residues ; analysis

Abstract:By using PVX derived vector pGR107, the effect of BYDV-MP nuclear localization signal on the movement of PVX was studied. BYDV-MP was cloned into pGR107 using GFP as an indicator. BYDV-MP was then shown to induce the systemic infection and exacerbate the symptom of PVX through infecting Nicotiana benthamiana. When the PVX gene encoding 25kD protein, which functioned as a systematic movemnet protein,was deleted and the above experiment was repeated, the result showed that BYDV-MP could compensate the systemic movement of PVX. A serial mutants with substitutions on the fifth, sixth and seventh amino acids of BYDV-MP nuclear localization signal was further constructed. It was found that the mutants at the fifth, sixth amino acids in BYDV-MP nuclear localization signal could only delay or weaken systemic movement of PVX whereas the mutant at seventh amino acid could entirely inhibit systemic movement of PVX.
Amino Acid Sequence ; Green Fluorescent Proteins ; genetics ; Luteovirus ; physiology ; Molecular Sequence Data ; Nuclear Localization Signals ; chemistry ; physiology ; Plant Viral Movement Proteins ; physiology ; Potexvirus ; genetics ; physiology

OBJECTIVETo discuss the optimal immunization dose by observing the immunoprotective effects of different doses of recombinant Schistosoma japonicum (Chinese strain) signaling protein 14-3-3 (rSj14-3-3).

METHODSSj14-3-3 gene was amplified by reverse transcriptase PCR (RT-PCR), subcloned into prokaryotic expression vector pET28a, then transformed into E.coli to express by inducing. Purified rSj14-3-3 was prepared through SDS polyacrylamide gel electrophoresis (SDS-PAGE), electroelution, dialysis, then BALB/c mice were divided into 5 groups and immunized in rSj14-3-3 protein followed by challenging infection (the 1st, 2nd, and 3rd groups were immunized in 50 microg, 100 microg and 300 microg antigen, respectively. The 4th, 5th groups were immunized in Freund's adjuvant and normal saline controls). After 6 weeks of challenging infection, the mice were killed and the worm and egg reduction rates were calculated. And the mice sera in different time were taken to examine the specific anti-Sj14-3-3 IgG.

RESULTSrSj14-3-3 protein was expressed successfully. After immunizing and challenging, worm reduction was found to be 28.20% in the 1st group, 43.10% in the 2nd group, 40.00% in the 3rd group, respectively. Number of eggs in liver tissue was reduced by 41.80%, 57.50%, 55.70%, respectively. Compared the results of the tested groups to the controls, the differences were of significance by t-test (worm reduction rate: t = 6.8 in the 1st group, t = 8.7 in the 2nd group, t = 7.3 in the 3rd group, P < 0.01 in all tested groups. Egg reduction rate at the group's number above: t = 11.23, t = 11.54, t = 7.99, P < 0.01 in all tested groups). As compared the results between the tested groups by chi(2), the differences were of significance between the 1st and the 2nd groups (worm reduction rate: chi(2) = 8.96, P < 0.05; egg reduction rate: chi(2) = 15.69, P < 0.05), between the 1st and the 3rd groups, the differences were also of significance (worm reduction rate: chi(2) = 6.52, P < 0.05; egg reduction rate: chi(2) = 12.52, P < 0.05). The difference was not of significance between the 2nd and the 3rd groups (worm reduction rate: chi(2) = 1.20, P > 0.05; egg reduction rate: chi(2) = 0.93, P > 0.05). In all tested groups, total anti-Sj14-3-3 specific IgG rose markedly. IgG(1) and IgG(2a) subtypes were high, but IgG(2b) and IgG(3) were near the background in four subtypes tested.

CONCLUSIONImmunoprotection of rSj14-3-3 should have some relations with immunization dose, and the protection obtained from immunizing mice by using 100 microg antigen was the best.

14-3-3 Proteins ; administration & dosage ; immunology ; Animals ; Antibodies, Helminth ; immunology ; Antibody Formation ; Antigens, Helminth ; blood ; Female ; Helminth Proteins ; immunology ; Immunoglobulin G ; blood ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; Schistosoma japonicum ; genetics ; immunology ; Signal Transduction ; Vaccination

OBJECTIVEA reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for rapid visual detection of HIV-1.

METHODSRT-LAMP primers were designed according to conservative sequences of HIV-1 gag gene, and their sensitivity and specificity were evaluated by the established RT-LAMP protocol with the addition of the hydroxynaphthol blue (HNB) dye prior to amplification. The performance of RT-LAMP on clinical samples was compared with real-time reverse transcription PCR(qRT-PCR).

RESULTSThe RT-LAMP assay showed a high specificity, and its detection limit was 1000 copies RNA per tube. The sensitivity and specificity of this method using 43 clinical samples were 94.6% and 100%, respectively,in comparison with those of qRT-PCR.

CONCLUSIONRT-LAMP assay using hydroxynaphthol blue dye does not need expensive instruments, and offer an alternative for the rapid detection of HIV-1 with the potential to be applied in field diagnosis.

HIV-1 ; isolation & purification ; Naphthalenesulfonates ; Nucleic Acid Amplification Techniques ; methods ; Real-Time Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction ; Reverse Transcription ; Sensitivity and Specificity

Laser-induced breakdown spectroscopy (LIBS) was used for analysis of the distribution of S,Mn,Fe,Cr,Mo,Si,Al in a 34CrNiMo6 steel sample cut from a main shaft of wind driven generator.The MnS inclusion area in each ablation craters cover zone was extracted in the way of comparing the metallograph captured by optical microscopy before and after LIBS scanning ablation.The statistic relation between MnS inclusion area and signal intensity of S and Mn was analyzed.The result showed that the abnormal signal of S and Mn occurred at the same position with the existence of MnS inclusion,and their signal intensity showed linear relationship.The abnormal signal of S and Mn were triggered mainly by MnS inclusion.The statistic result also showed linear relationship between signal intensity and MnS inclusion area both for S and Mn.It was possible to determine the inclusion type,size and distribution by analyzing abnormal signal.A simplified ablation model was established to calculate the relation of S and Mn content to MnS inclusion area.The arithmetic result showed a linear relation between the content and MnS inclusion area both for S and Mn.The calculation confirmed the linear relationship between signal intensity and inclusion area observed in experiment.The linear relationship could be interfered by macro-segregation,micro-segregation,deviation in measuring inclusion area,and inclusion spatter in pre-ablation.