Objective To study the value of the semiquantitative-parameter analysis of wash out index of time-intensity curve (Swash-out) in evaluating the therapeutic effect of neoadjuvant chemotherapy for locally advanced breast cancer (LABC).Methods Fifty-nine women with LABC underwent dynamic contrast enhancedt MRI examination before chemotherapy,after the 2nd cycle and the 4th cycle of chemotherapy.All patients were divided into major histological response group (MHR) and non-major histological response group (NMHR) according to the final pathologic response.Swash-out and the variancetrends of Swash-out before NAC,after the 2nd cycle of NAC and after the 4th cycle of NAC were compared in each group and between the two groups.According to the gold standard of Miller & Payne criterion,Receiver operating characteristic curve (ROC) analysis was performed to evaluate the predicting effect of Swash-out for NAC response,and to compare it with Semi-quantitative TIC curve indicators Smax (steepest slope) and PPE (peak percent enhancement).Results Fifty-nine patients of LABC patients were divided into a MHR group of 34 patients and a NMHR group of 25 patients.Swash before NAC of MHR group was-16.99 (-56.72-41.20),Swash-out after the 2nd cycle of NAC was 5.66(-69.45-53.08),Swash-out after 4th cycle of NAC was 15.95 (-7.80-54.23).Swash-out before NAC of NMHR group was-23.08 (-64.24-34.39),Swash-out after the 2nd cycle of NAC of NMHR group was-23.01 (-52.72-28.70),Swash-out after 4th cycle of NAC of NMHR group was-11.45 (-50.49-50.93).Swash-out variance rate of MHR group after the 2nd and the 4th cycle of NAC were-1.18 (-31.32-60.86) and 1.50 (-86.27-3.61),respectively.Swash-out variance rate of NMHR group after the 2nd and the 4th cycle of NAC were-0.28(-3.24-9.46) and 0.27 (-5.34-3.11),respectively.Swash-out was not significantly different between the two groups before NAC (Z =-0.97,P ＞0.05).Swash-out and Swash-out variance rate of MHR group after the 2nd cycle of NAC were significant higher than that of NMHR group (Z =-3.97 and-3.02,P ＜0.01).Swash-out and Swash-out variance rate of MHR group after the 4th cycle of NAC were significant higher than that of NMHR group (Z =-3.96 and-3.16,P ＜ 0.01).Area under curve (Az) after the 2nd and the 4th cycle of NAC were 0.805 and 0.804,respectively,and no significant difference was found between them (Z =0.019,P ＞0.05).Diagnostic cut-off points were-8.670 for the 2nd cycle of NAC and 4.105 for the 4th cycle of NAC.Diagnostic sensitivity was 79.42％,specificity was 76.00％ and Youden index was 0.554,for after the 2nd and the 4th cycle of NAC.Conclusion Swash-out of TIC curve before NAC cannot predict the response of NAC,Swash-out of TIC curve after the 2nd cycle of NAC and after the 4th cycle of NAC are efficient in predicting the response of NAC.

Objective To develop the method for simultaneous identification and quantification of morphine,6-monoactylmorphine,codeine,heroin,ketamine,3,4-Methylenedioxyamphetamine (MDA),3,4-methylenedioxymethamphetamine (MDMA),amphetamine and methylamphetamine in human plasma.Methods UPLC-MS/MS was adopted to analyze plasma with protein precipitated using 10％ trichloroacetic acid.Plasma samples were separated on ACQUITY UPLC HSS C18 column with aqueous solution (0.01％ formic acid)-methanol (0.01％ formic acid) as the mobile phase,and at a flow rate of 0.3 mL·min-1.The multiple reaction monitoring (MRM) mode was performed combined with the positive ion mode for quantification in four sections.Results The retention time,the characteristic fragment ions,and the relative abundance ratio of the molecular ion peak could be used to identify these nine compounds sensitively.The liner calibration curve of morphine,codeine,heroin,ketamine,6-monoacetylmorphine,MDA,MDMA,amphetamine and methamphetamine were obtained in the concentration range of 5.00×10-3～5.00 (r=0.9934),1.00× 10-2～ 10.00 (r=0.9905),1.00× 10-2～ 10.00 (r=0.9929),2.50× 10-3 ～2.50 (r=0.9960),5.00× 10-3 ～ 5.00 (r=0.9925),5.00× 10-4 ～ 5.00 (r=0.9910),5.00× 10-4 ～ 5.00 (r=0.9924),5.00× 10-4 ～ 5.00 (r=0.9920) and 5.00×10-4～5.00 μg·mL-1 (r=0.9900) respectively.The lowest detection limit was 1.00,1.00,1.00,0.50,0.50,0.10,0.10,0.10 and 0.10 ng· mL-1 respectively.The relative standard deviation (RSD) of the inter-day and intra-day were less than 18％.The relative recovery was more than 60％,and the RSD were less than 15％.Conclusion The method is accurate,sensitive and suitable for identification and quantification of 9 drugs of opiates,ketamine and amphetamines in human plasma.

The fumarase from the culture of Aspergillus wentii F-871 was partially by means of precipitation with protamine sulphate, ammonium sulfate fractionation and column chromatography on Sephadex G-200, and then the concentrated enzyme solution was freeze-dried. The comparative enzyme activity of the fumarase was increased by 31.70 times and reached 24.6 U/mg, and the recovery was 36.64% . The optimal pH and temperature was 8.0 and 30℃, respectively. The pH stability of fumarase ranged 6.0 - 8.5, and more than 90% of the enzyme activity remained after incubated at 35℃ for 30 min.

Purpose To explore the diagnostic value of double exponential model for pelvic lesions using 3.0T MRI for the diagnosis of pelvic lesion. Materials and Methods Fifty patients with pelvic lesions (30 benign cases and 20 malignant cases) underwent MR750-diffusion weighted imaging (DWI) scans, with b values of 0, 50, 300, 600, 800 and 1200 s/mm2, Functool-MADC software was used on AW 451 workstations for data processing, Slow ADC value, Fast ADC value, Standard ADC value, Fraction of fast ADC value were recorded and compared between benign and malignant lesions, and Standard ADC images were fused with axial T2 fat-suppressed images. Results Slow ADC values [(1.83±0.86)×10-3 mm2/s] and Standard ADC values [(1.79±0.78)×10-3 mm2/s] of benign lesions were larger than those of the malignant lesions [Slow ADC values:(1.05±0.31)×10-3 mm2/s;Standard ADC values:(1.13±0.39)×10-3 mm2/s] (t=3.90, 3.51;P<0.01), and the difference of Slow ADC value was largest between benign and malignant lesions. Slow ADC values of both benign and malignant lesions were significantly less than the Fast ADC values [benign:Slow ADC value=(1.83±0.86)×10-3 mm2/s, Fast ADC value=(16.95±8.63)×10-3 mm2/s; malignant: Slow ADC value=(1.05±0.31)×10-3 mm2/s, Fast ADC value=(15.12±9.90)×10-3 mm2/s] (t=-10.40,-6.29;P<0.01). Conclusion Double exponential decay model is capable of differentiating benign and malignant pelvic tumors, thus is of great significance for clinical preoperative diagnosis.

Objective To investigate the changes of serum cardiac troponin I(cTnI)and brain natriuretic peptide(BNP)in heart failure of children with pneumonia and their relationship with heart function.Methods Thirty healthy children aged from 5 months to 3 years old were randomly selected with 17 male and 13 female(healthy group).Thirty children with severe heart failure aged from 3 months to 2 years old were selected at the same time with 21 male and 9 female(heart failure group).Thirty children with ordinary pneumonia aged from 3 months to 3 years old were also sampled with 16 male and 14 female(ordinary pneumonia group).The peripheral bloods of 2-3 mL of all children were taken.The BNP level were measured by enzyme-linked immunosorbent assay and the cTnI level was determined by micro-particle enzyme immunoluminescent.Left ventricular ejection fraction(LVEF) and left ventricular fractional shor-tening(LVFS)were detected by echocardiography.SPSS 11.0 software was used to analyze the data.Results The levels of cTnI [(0.389?0.030) ?g/L] and BNP [(0.572?0.090) ?g/L] of heart failure group increased significantly compared with healthy and ordinary pneumonia group,while their LVEF and LVFS decreased significantly(Pa

The implementation of knowledge management (KM) in hospitals affects efficiency and outcomes of hospitals.However,few studies explored the implementation of KM in China.Twenty-two questions were designed concerning KM implementation status in over 50 hospitals.In order to understand the KM level and attitude to KM of the hospital's managers,a random sampling survey was conducted among 138 managers from 50 different scales of hospitals in 15 provinces of China.The survey showed that overall level of KM implementation in Chinese hospitals was still low and differed among different scales of hospitals (P＜0.05,or P＜0.01).In all the hospitals investigated,63.8％ did not implement KM yet,among which 46％ even had not planned for that.49.8％ of the hospitals investigated had no training program about KM ever and the main source of hospital staff to get knowledge was iuternet.It suggested that hospital managers should make much more efforts to get to know and understand theories on KM,so that hospital KM could be promoted more rapidly.

The optimum condition of shaking-flask p roducing enzyme were the tempe rature 26℃,initial pH 6 4,fermentation period 19 hours,medium volume 15mL m e dium/300mL Flask.soymilk-clotting enzyme was obtained from ammonium sulfate p r ecipitation.The optimum temperature and pH for the soymilk-clotting activity wa s 70℃and 5 8.The enzyme was easy to lose activity in acid or alkaline circumst a nce.About 60% of the original activity remained after 1 hour at 60℃.Ca 2+ ,Fe 2+ , Mg 2+ ,Na +increased the clotting activity,whereas Zn 2+ ,Al 3+ ca use inhibition.

The alkaline lipase gene of Penicillium expansum (PEL) was coloned into the yeast integrative plasmid pPIC3.5K, which was then transformed into His4 mutant yeast GS115. Recombinant Pichia strains were obtained by minimal olive oil-methanol plates screening and confirmed by PCR. The expression producus of PEL gene was analysis by SDS-PAGE and olive oil plate, the result indicated that PEL gene was functionally overexpressed in Pichia pastoris and up to 95% of the secreted protein. Recombinant lipase had a molecular mass of 28kD, showing a range similar to that of PEL, could hydrolyze olive oil and formed clear halos in the olive oil plates. Four different strategies (different media, pH, glycerol and methanol concentration) were applied to optimize the cultivation conditions, the activity of lipase was up to 260 u/mL under the optimal cultivation conditions. It is pointed out that the absence of the expensive biotin and yeast nitrogen base in the medium increased the lipase production. The possible reason of this result is absence of yeast nitrogen base increased the medium pH during cultivation, and PEL shows a higher stability at this condition. The lipase activity of the supernatant from the culture grown at pH 7 was higher than the one from the culture in the same medium at pH 6.0 is due to the pH stability of PEL too. The results also showed that the methanol and glycerol concentration had a marked effect on the production of lipase.
Enzyme Stability ; Fungal Proteins ; genetics ; metabolism ; Genetic Vectors ; genetics ; Hydrogen-Ion Concentration ; Lipase ; genetics ; metabolism ; Models, Genetic ; Penicillium ; enzymology ; genetics ; Pichia ; genetics ; metabolism ; Plasmids ; genetics

In the present study, the genome shuffling was used to improve lipase production of Penicillium expansum. A lipase producing mutant strain-Penicillium expansum FS8486 and a wild type of Aspergillus Tamarii FS-132 isolated from soil of a volcano in Xinjiang were used as the parental strains. After two rounds of genome shuffling, several elite daughter strains were screened. The lipase activity in one of the daughter strains was increased 317% over the starting strain FS8486. Comparisons of the morphology, RAPD (Random Amplification of Polymorphic DNA) polymorphism and the fatty acid compositions between the daughter and the parental strains suggested that the filial generation were generated by genome shuffling. In this study, the genome shuffling used successfully first time in eukaryotic microorganism and increases the production of the desired metabolite in short time, the study will be useful to spread the genome shuffling in eukaryotic microbial breeding.
Aspergillus ; genetics ; DNA Shuffling ; methods ; Genetic Enhancement ; methods ; Genome, Fungal ; genetics ; Lipase ; biosynthesis ; genetics ; Penicillium ; enzymology ; genetics ; Random Amplified Polymorphic DNA Technique

OBJECTIVETo evaluate the impact of specific siRNA on survivin gene in transfected leukemia cells.

METHODThe small interfering RNA (siRNA) targeted survivin mRNA was synthesized in vitro and was transfected into K562 cell by Hiperfect into human leukemia cell line K562, which has high survivin expression level. The level of survivin mRNA expression was determined by quantitative reverse transcription polymerase chain reaction (RT-PCR) with SYBR GREEN I. The apoptosis index of cytotrophoblasts were determined and analyzed by FCM (Annexin V-FITC/PI staining methods). The cell proliferation was examined by MTT at 48 h and 72 h after transfection.

RESULTThe level of mRNA expression was significantly inhibited by the siRNA 48 h and 72 h after transfection, the suppression rate of survivin mRNA separately reached 85.21%, 94.35% mensurated by quantitative RT-PCR with SYBR GREEN I, cell proliferation was inhibited significantly by 45.02% and 50.88%, respectively, the apoptotic rate detected by Annexin V-FITC assay reached 12.28%and 21.55%, respectively.

CONCLUSIONThe chemosynthesized siRNA targeting survivin could significantly down-regulate survivin mRNA. Survivin siRNA was able to inhibit the proliferation of leukemia cell line K562. Survivin may become a new target for leukemia gene therapy.

Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Gene Silencing ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; K562 Cells ; RNA, Small Interfering ; pharmacology ; Transfection