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To determine a dengue fever outbreak in Yiwu city, Zhejiang Province in 2009 and to trace the origin of the pathogen.The dengue virus IgM, IgG antibodies and viral nucleic acid were detected and virus was isolated using 40 serum samples from the suspected patients. The viral RNA of the isolated virus strains was extracted and the E gene was amplified by RT-PCR. The amplicons were sequenced and the phylogenetic and homological analyses were also constructed.Among 40 serum samples from dengue fever suspected patients, 17 were positive from for dengue IgM (42.5%); 4 were IgG positive (10.0%); 34 samples were dengue virus RNA positive (85.0%), 28 dengue virus type 3 (D3) strains were isolated (70.0%). The complete coding region of envelope genes (E) from 13 D3 strains was all 1479 nt without any insertion or deletion, which encoded with 493 amino acids (aa). E gene from the 13 D3 strains from Zhejiang in 2009 (D3/ZJ/2009) was 100.0% identical. The strain from Saudi Arabia shared the highest similarity with the D3 strain, 99.3% and 100.0% of their E genes and deduced amino acids were identical, respectively, whereas they were 93.4% and 97.4% between D3/ZJ/2009 strain and its prototype strain (D3/H87/1956), and 93.6% and 97.4% between D3/ZJ/2009 and a D3 strain isolated in Guangxi Province in 1980. The phylogenetic tree of E genes also indicated that D3/ZJ/2009 had maximum similarity with the D3/Saudi Arabia/2004. They all belonged to the D3/GIII branch, which was originated from Indian Subcontinent.The outbreak of dengue fever in Zhejiang in 2009 was caused by type 3 dengue virus III genotype. The virus was most likely originated from Saudi Arabia.

to evaluate the diagnostic accuracy of ultrasound-guided core needle biopsy of breast tumors.six hundred and sixty-seven cases of core needle biopsy of breast encountered during the period from January, 2004 to June, 2007 were retrieved from the archival file and retrospectively reviewed. The core needle biopsy diagnoses were correlated with the histologic findings of the subsequent surgical excision specimens. The discrepancies were further analyzed.three hundred and eighty-two patients had core needle biopsy diagnosis followed by local excision, breast conservation surgery or mastectomy. Two hundred and eighty-one cases were confirmed to have malignancy in the surgical specimens. Review of the corresponding core needle biopsies showed 4 false-negative cases, no false-positive cases, 28 cases with underestimation and 2 cases with overestimation. The false-negative rate was 1.4% (4/281). The rate of underestimation for ductal carcinoma-in-situ was 6/11. The diagnostic accuracy of core needle biopsy was 94.7% (266/281).in order to improve the diagnostic accuracy of core needle biopsy of breast tumors, recognition of the limitation of the procedure, application of immunohistochemistry and awareness of potentially rare entities are important.

The purpose of this study was to evaluate the importance of diffusion-weighted magnetic resonance imaging (DW-MRI) apparent diffusion coefficient (ADC) values to predict treatment response to neoadjuvant chemotherapy (NCT) in patients with locally advanced breast cancer (LABC). Thirty-two patients with LABC underwent 2–4 cylces of NCT (docetaxel and epirubicin). The DW-MRI scans were performed within one week prior to chemotherapy and after the first course of treatment, respectively. Accordingly, tumor volumes, changes in tumor ADC values, and their degree of correlation were analyzed. The overall response (OR) was observed in 62.5% (95% CI, 45.7–79.3%) of patients after 2 cycles of NCT. The clinical complete response (CR) rate and pathological CR (pCR) rate were 15.6 and 9.4%, respectively. The stable disease (SD) rate was 34.4% (11 patients), and progressive disease (PD) was observed in only one patient (3.1%). After the first cycle of NCT, the ADC values in the CR + PR group significantly increased (P

To screen specifically expressed genes in the development of nerve, muscle, and body axis of amphioxus, Branchiostoma belcheri tsingtauenese.A subtractive cDNA library was constructed from the 12-hour amphioxus neurula cDNA after subtractively hybridized with the 6-hour amphioxus gastrula cDNA. The total RNA was extracted from the 12-hour neurula and 6-hour gastrula, then reverse transcribed into cDNA. The 12-hour neurula cDNA was designated as the experimental group (the tester) and the 6-hour gastrula cDNA as the control group (the driver). The differentially expressed sequences were exponentially amplified using suppression PCR. Background was subtracted and differentially expressed sequences were further enriched. The PCR products were ligated to the T Vector. After transformation of the recombinant plasmid carrying inserted amphioxus cDNA into E.coli host cells, the cDNA library was constructed successfully.Two hundred randomly chosen positive clones were sequenced and some of neurula-specifically expressed genes were obtained.SSH is an effective method for searching differentially expressed genes. The subtractive cDNA library we generated provides a tool for further study of regulatory mechanisms of amphioxus early embryonic development.

Microwave cavity is the core part to the physics package of hydrogen maser. The most suitable mode to achieve hydrogen maser self-oscillation was considered and dimensions of the cavity with 1.42G cavity frequency in TE011mode were calculated. The calculated result of the cavity was confirmed by FEM simulation. Then the factors which may affect the stability of Hydrogen maser were analysed. Finally, the maser oscillation signal was obtained and the stability of the signal was tested.

Equivalent sliding mode control based on RBF neural network uses the traditional gradient descent algorithm to achieve the control function. Because of local minima, training is slow and so on. The algorithm has slow convergence, poor adaptability problems. This paper presents a RBF network based on variable learning rate of W which can be used to equivalent sliding mode control system. Experimental results of the simulation show that the new algorithm has fast convergence and tracking precision. It can effectively avoid the interference caused by unknown divergence, and have a good control of reliability.

Background The 6986A > G polymorphism for CYP3A5 and the − 129T > C , 1236C > T , 2677G > T/A , and 3435C > T polymorphisms for MDR1 are considered the major genetic factors affecting a range of drugs' metabolism and transport. Simultaneous genotyping of these five polymorphisms would be useful for estimating the therapeutic effects of their related drugs. Subjects and methods We have described a SNaPshot assay that can simultaneously detect all the five polymorphisms based on multiplex PCR and minisequencing reaction. A total of 168 unrelated Chinese DNA samples were used to establish and evaluate the assay. Results The different genotypes of the five polymorphisms could be determined by peak retention time and colors. DNA sequencing was performed on samples randomly selected from each of the genotype groups detected by SNaPshot assay, and the results indicated 100% concordance. Conclusion The SNaPshot assay for the CYP3A5 and MDR1 five polymorphisms detection was accurate, automated, and cost-effective.

In this paper, the validity of the near-field equivalent method is studied to solve the problem of distortion in simulations. To analyze the validity, the condition of the principle of equivalence is reconsidered. The relationship between the principle of equivalence and the methods of near-field equivalence and far-field equivalence is studied, results show that the distance between the antenna and its platform is the key factor affecting the validity. To verify it, the 2D radiation patterns of the three-knife antenna above the curved platform are analyzed with full modeling simulation method and near-field equivalent method, respectively. Results indicate that the near-field equivalent method can still be used to simulate the radiation patterns of antennas above the platform, but the accuracy of this method is closely related to the distance between the antenna and its platform. That is, the longer the distance is, the more accurate the results are; when the distance is longer than 1/3 wavelength, high-precision results can be achieved with near-field equivalent method; However, when the distance is shorter than about 1/30 wavelength, this method is no longer appropriate for the simulation.

The ion channels located on the cell fine structures play an important role in the physiological functions of cell membrane. However, it is impossible to achieve precise positioning on the nanometer scale cellular microstructures by conventional patch-clamp technique, due to the 200 nm resolution limit of optical microscope. To solve this problem, we have established a high-resolution patch-clamp technique, which combined commercial scanning ion conductance microscopy (SICM) and patch-clamp recording through a nanopipette probe, based on SICM feedback control. MDCK cells were used as observation object to test the capability of the technique. Firstly, a feedback controlled SICM nanopipette (approximately 150 MOmega) non-contactly scanned over a selected area of living MDCK cells monolayer to obtain high-resolution topographic images of microvilli and tight-junction microstructures on the MDCK cells monolayer. Secondly, the same nanopipette was non-contactly moved and precisely positioned over the microvilli or tight-junction microstructure under SICM feedback control. Finally, the SICM feedback control was switched off, the nanopipette slowly contacted with the cell membrane to get a patch-clamp giga-ohm sealing in the cell-attached patch-clamp configuration, and then performed ion channel recording as a normal patch-clamp electrode. The ion channel recordings showed that ion channels of microvilli microstructure opened at pipette holding potential of -100, -60, -40, 0, +40, +60, +100 mV (n=11). However, the opening of ion channels of tight-junction microstructure was not detected at pipette holding potential of -100, -40, 0, +40, +100 mV (n=9). These results suggest that our high-resolution patch-clamp technique can achieve accurate nanopipette positioning and nanometer scale high-resolution patch-clamp recording, which may provide a powerful tool to study the spatial distribution and functions of ion channel in the nanometer scale microstructures of living biological samples.

Heparan sulfate proteoglycans (HSPGs) are involved in the regulation of cell growth, apoptosis and lipid metabolism in vitro. Syndecans are the primary form of HSPGs. Syndecan-1 is involved in the processes of cell growth, differentiation, adhesion, wound healing and inflammation. Additionally, as a sinusoidal transmembrane HSPG facing the plasma compartment, syndecan-1 is a promising target to be involved in lipoprotein physiology. We aimed to examine the possible correlation of syndecan-1 and lipid profile in type 2 diabetes mellitus. In this study, serum syndecan-1 was detected by ELISA, and potential correlations between syndecan-1 and triglyceride, cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, lipoprotein a, apolipoprotein (apo) B, apoA1 and apoB/apoA1 were analyzed. Forty-one patients with type 2 diabetes and 31 age-matched, non-diabetic healthy subjects (controls) were enrolled. Syndecan-1 in patients with diabetes (26.15 ± 2.42 ng/ml) was significantly higher than that of the controls (16.85 ± 1.98 ng/ml, t = −2.98, P = 0.005). Serum syndecan-1 level correlated negatively with apoA1 (r = −0.46, P = 0.003). Multiple regression analysis showed that apoA1 (b = −0.43, P = 0.003) was a predictor of serum syndecan-1 levels in subjects with type 2 diabetes.

Magnetic state selector is a key component in hydrogen atom maser. Its design and improvement play important role in the H maser's working parameters, volume and weight. This paper primarily introduces the principle of 6-pole and 4-pole magnetic selector. We also show the optimal size of team optics and their experiment test comparison. Nowadays, these have been successfully used in atomic hydrogen maser according to their characteristics.

Support from the Chinese Academy of Sciences and the General Armament Department, certain technology improvement and enhancement of atomic hydrogen clock was effectively carried out by Shanghai Astronomical Observatory under the promotion of BD, lunar exploration, deep space tracking projects. The atomic hydrogen clock's reliability and performance have been greatly improved. The developed atomic hydrogen clocks play an important role in some national scientific experiments and military projects. Performance evolution shows that the active hydrogen clock's stability is better than 2E-15/day, temperature coefficient is better than 5 E-15/°C.

To evaluate the effects and toxicity of the neoadjuvant chemotherapy of docetaxel combined with epirubicin or pirarubicin on breast cancer, and to investigate the influencing factors of the response to neoadjuvant chemotherapy.160 patients with stage II/III breast cancer, all females, aged 47 (22-66), were treated with docetaxel plus epirubicin or pirarubicin with 3 weeks as a cycle. Two to six cycles of treatment were given before surgery. The clinical efficacy and toxicity of the treatment were evaluated, and the correlation between the influencing factors and the clinical parameters with treatment response was analyzed.The clinical response rate (RR) was 90% (144/160), the complete response (CR) rate was 26% (41/160), the partial response (PR) rate was 64% (103/160). The stable disease (SD) rate was 8% (13/160). The progress disease (PD) rate was 2% (3/160), the pathologically complete remission (pCR) rate was 7% (11/160), and the tumor-pathological complete response (tpCR) rate was 2% (1.3/160). Univariate analysis showed that the tumor size, clinical stage, triple negative phenotype might be the meaningful parameters influencing the clinical response. The patients with smaller tumor size, low stage tumor, and being triple-negative were more likely to achieve CR (P = 0.0371, 0.0013, and 0.0019 respectively). Age, histological grading, ER/PR ratio, Her-2 status did not significantly influence the early response. Multivariate analysis showed that the disease stage might be the meaningful factors for better response (P = 0.0030). The major toxic reactions of the therapy included neutropenia, alopecia, nausea, and vomiting.The combination neoadjuvant chemotherapy with docetaxel and epirubicin or pirarubicin is an effective method to treat breast cancer with tolerable toxicity. The meaningful parameter influencing the early response is clinical stage.

Four hydrogen masers have being put into operation at the earth station of Chinese positioning system for more than six years. We have made many physical and electronics improvements on these hydrogen masers for improving the long-term performance. The hydrogen maser's size is 110cm(high)*54cm(width)*73cm(depth). It weighs 180kg. By performance comparing with each other, we may choose the best one as a frequency reference for this system. A kind of cavity auto-tuning system was applied for improving of the long-term performance. Now another 13 H masers were prepared for the Compass-2 system. Performance evaluation showed the frequency stability is in the order of E-15 for time interval from 100s to 1day and the phase noise (SSB) is −150 dBc/100Hz. The environmental sensitivities are also given in this paper.

To study the synergistic mechanism between PH II -7 and doxorubicin against multi-drug resistant HL-60/ADR cells and its parent HL-60 cells.The anti-tumor activity of doxorubicin alone and combined with PH II -7 were measured by MTT assay. RNA was extracted from the cells treated with PH II -7 for different times or doses then the expression of MRP gene was measured by RT-PCR. Confocal laser scanning microscopy and FACS were used to detect the intracellular cumulation of doxorubicin in PH II -7 treated HL-60 and HL-60/ADR cells.PH II -7 has anti-tumor effect with IC50 of (0.83 +/- 0.08) micromol/L and (1.74 +/- 0.56) micromol/L for HL-60 and HL-60/ADR, respectively. It could potentiate the anti-tumor effect of doxorubicin with CDI of 0.7 and 0.43 for HL-60 and HL-60/ADR, respectively. PH II -7 and doxorubicin act synergistically in inhibiting the proliferation of HL-60 and HL-60/ADR cells and down-regulating the expression of MRP gene in a dose and time dependent manner. PH II -7 restored the intracellular cumulation of doxorubicin in HL-60/ADR cells to 55% of that in HL-60 cells.PH II -7 can significantly hasten the cytotoxicity of doxorubicin to HL-60 and HL-60/ADR cells through down-regulating the expression of MRP. The synergistic effect was more obvious in HL-60/ADR cells.

Three new alkaloids-griffithazanone A (1), griffithdione (2), and griffithinam (3)-were isolated from the roots of Goniothalamus griffithii, along with six known compounds, 4-methyl-2,9, 10-(2H)-1-azaanthracenetrione (4), velutinam, aristololactam BI, aristololactam BII, aristololactam AII, and norcepharanone B. Their structures were elucidated on the basis of spectral and chemical methods. The absolute configuration of griffithazanone A (1) was determined by the preparation of Mosher's esters.

To investigate and analyze the traumatic conditions of Longnan area of Gansu in Wenchuan earthquake, and to explore the measures coping with unexpected disasters.A sample survey including questionnaire survey, interviews and other methods was adopted to investigate the conditions of affected population of Longnan area in earthquake, especially on the traumatic conditions, the injured causes, treatment methods.In the sample survey of a total of 1652 earthquake casualties, the dead were 16 and the patients with extremities fractures were 750 (45.4%). Organized and timely rescue of the wounded could reduce the complications and the mortality .Strengthening the prevention and treatment of earthquake trauma can effectively decrease the death rate and disabled rate.

To investigate the contamination of Staphylococcus aureus isolates in ice cream by phenotypic typing and molecular typing.The Staphylococcus aureus isolates were separated from ice cream, filler, cutter, salves and material. The separated isolates were characterized by drug-resistance, staphylococcal enterotoxin (SEA-E), SE (A-E, G-J) genes and pulsed-field gel electrophoresis (PFGE) types.Two Staphylococcus aureus isolates were separated, one from ice cream, another from cutter. Their characteristics of drug-resistance, staphylococcal enterotoxin (SEA-E), SE (A-E,G-J) genes and PFGE type were the same.The two Staphylococcus aureus isolates were the same clone. The contaminated Staphylococcus aureus isolates could be traced to the contaminated cutters.

Objective To observe the ultrastructure of human oocytes at different maturity stages and morphologies before and after in vitro maturation. Design Case report. Setting University-based center for human reproduction. Patient(s) A 28-year-old patient with polycystic ovary syndrome undergoing IVM/IVF-ET treatment. Intervention(s) Human chorionic gonadotropin was given to the patient 36 hours before her oocytes were retrieved by transvaginal aspiration. Main Outcome Measure(s) Ultrastructure of human oocytes matured in vitro. Result(s) Obvious ultrastructural differences were found in oocytes at different maturity stages between those cultured in vitro and those matured in vivo. Degenerate structures were observed in abnormal oocytes. Granulosa cell apoptosis rates from abnormal oocytes were higher than those from normal oocytes. Cumulus cells surrounding morphologically abnormal oocytes had a significantly higher frequency of cell apoptosis, as compared with morphologically normal oocytes. Conclusion(s) Obvious ultrastructural differences were found between the oocytes matured in vitro and in vivo.

VRML is a very powerful language for displaying 3D objects and supporting interactivity, sophisticated lighting and cinematography. For reasons of processing speed and availability of 3D libraries, formats of VRML viewers are generally implemented in native C/C++ code. However, these languages may not be the most elegant languages to use for artificially intelligent control of 3D for dramatic narrative, believable speaking characters, dynamic cinematography and stage management. A good alternative is the Java language which provides very productive syntax and a body of developed application code for control and management of 3D scenes. The paper first presents an analysis of the problems of interfacing Java to 3D rendering systems, and some possible solutions in the context of interfacing Java to VRML. Finally we implement a Java interface which is defined independent to the underlying 3D representation and format. This solution makes it possible to abstract out the particular field types used in the 3D rendering platform. Therefore a Java control system using the interface presented here should be simple to port and integrate with any 3D rendering system using any 3D format.

Sensor positioning is a fundamental and crucial issue for wireless ad-hoc local area network operation and management. In this paper, the hierarchical maximum likelihood algorithm for determining the positions of nodes is presented in details. The numerical simulation of the hierarchical maximum likelihood algorithm shows that with no path-loss fluctuation, the localization error decreases with increasing grid resolution increment step eta; as the path-loss fluctuation increases, the advantages of using larger eta value become weaker; and with very big path-loss fluctuation, the performance of the pure maximum likelihood and the hierarchical maximum likelihood algorithm with different eta values becomes comparable.

To construct and express human 4-1BBL/anti-CD20 bispecific fusion protein and identify its biological activity.PCR and overlapping PCR were used to construct human 4-1BBL/anti-CD20 bispecific fusion protein. DNA sequencing was performed by the terminus of the fusion protein nucleotide.The product was purified by affinity chromatography and analyzed by Western blot and its antigen-binding activity was examined by FACS.The data of DNA sequence showed that human 4-1BBL/anti-CD20 bispecific fusion protein was correct. The fusion protein was recovered in high yield (up to 4 mg/L) after E-tag purification and predominantly(90%) as a dimer. The fusion protein could bind to Raji cells(CD20(+)) and A549 cells(4-1BB(+)), respectively.The human 4-1BBL/anti-CD20 bispecific fusion protein with high level expression was successfully obtained and could bind to Raji ceIls and A549 cells.

To evaluate the clinical efficacy and toxicity of combination neoadjuvant chemotherapy with docetaxel and pharmorubicin in the treatment of locally advanced breast cancer.From June 2005 to March 2007, 94 breast cancer patients who have been pathologically confirmed by core needle biopsy were treated with neoadjuvant chemotherapy before operation. Docetaxel 75 mg/m(2) plus pharmorubicin 80 mg/m(2) were administered as intravenous infusion on the first day of each 3-week cycle. Accepted 2 to 4 cycles of the treatment, the patients were underwent surgery after 12 to 16 days.The overall response rate (RR) was 80% (76/94). The complete clinical response rate (CR) was 22% (21/94). The partial response rate (PR) was 58% (55/94). The stable disease (SD) rate was 17% (16/94). The progress disease (PD) rate was 2% (2/94). The pathological complete rate (pCR) was 3% (3/94). The major toxic reactions of the therapy were neutropenia, alopecia, nausea and vomit. 76 patients (80%) suffered with grade 3 to 4 neutropenia. 4 patients suffered with grade 3 to 4 thrombocytopenia. 84 patients suffered with severe alopecia. 90 patients (95%) accepted supportive treatment of G-CSF. Septicemia and death were not occurred in this study.The combination neoadjuvant chemotherapy with docetaxel and pharmorubicin is an effective method to treat breast cancer and the toxicities are tolerable.

In our ongoing investigation of the bioactive constituents from plants, two new lignans, magnolignan A-2-O-beta-D-glucopyranoside and strebluslignanol were isolated from heartwood of Streblus asper, along with three known lignans, magnolignan A, magnolol, and magnaldehyde D. 1D and 2D NMR experiments, including COSY, HMQC, and HMBC, and other spectroscopic methods, including UV, IR, and MS were used for the determination of the structures and NMR assignments. Primary bioassays showed that magnolignan A-2-O-beta-D-glucopyranoside and strebluslignanol have medium cytotoxic activity against HEp-2 and HepG2 cells, with IC(50) of 13.3 microM, 46.4 microM and 10.1 microM, 21.7 microM, respectively.

To investigate the cell death pattern of sinusoidal endothelial cells (SECs) caused by ethanol and the effects of vascular endothelial growth factor (VEGF) on this cell death, as well as the underlying mechanism involving Ets-1 and Caspase-8.SECs were isolated from male Wistar rats and cultured in medium containing ethanol (25 - 100 mmol/L). VEGF (20 - 30 ng/ml) was added into the medium to be co-incubated for up to 6 h. Apoptosis was detected by terminal deoxynucleotidyl transferase (TdT)-mediated d-uridine triphosphate (dUTP)-biotin nick end labeling (TUNEL) technique. The protein expression of Ets-1, prototype of anti-apoptotic Ets family, was determined by Western blotting and the Caspase-8 was measured by FLICE/caspase-8 colorimetric protease assay kit.Three days after culture, the SECs showed spindle-like shapes and nearly confluent, however, the cells tended to shrink and die during the time course of ethanol incubation under phase contrast microscope. The control SECs contained only a few percent of TUNEL-positive cells; however, the TUNEL-positive cells started to increase 2 hours after the addition of ethanol (100 mmol/L), and about 75% of the cells were TUNEL-positive 6 hours after ethanol incubation (P0.05) under fluorescent microscope. Again, TUNEL-positive cells increased 6 hours after ethanol (25 - 100 mmol/L) incubation in a dose dependent manner (P0.05). Six hours after VEGF (20 - 30 ng/ml) was added into the medium with ethanol (100 mmol/L) the percentage of TUNEL positive SECs decreased in a dose dependent manner (P0.05), the level of ethanol-induced apoptotic cells in the presence of VEGF (30 ng/ml) being around 71% 6 hours after ethanol incubation alone. The Ets-1 protein level of the SECs decreased 6 hours after ethanol (100 mmol/L) incubation, which was prevented almost completely by VEGF (30 ng/ml). The Caspase-8 activity level was significantly increased to 44.9 +/- 14.3 2 hours after ethanol (100 mmol/L) incubation, and decrease to 30.4 +/- 2.0 and 25.2 +/- 2.2 respectively after the addition of VEGF (20 - 30 ng/ml) (both P0.05).VEGF prevents the apoptosis of primary cultured SECs induced by ethanol, through at least in part, inhibition of ethanol-induced down-regulation of Ets-1 protein expression and ethanol-induced up-regulation of Casepase-8 activity in SECs.

The differences in metabolism in Escherichia coli DH5alpha and its acetate-tolerant mutant DA19 were analyzed based on the activity of key enzymes involved in central metabolism when both strains were continuously cultured in nitrogen source-limited defined media. The activity of glucose-6-phosphate dehydrogenase (G6PDH) and isocitrate dehydrogenase (ICDH) in DA19 increased as compared with those in DH5alpha, while acetate kinase (ACK) and phosphofructosekinase (PFK) decreased. These indicated that more carbon flux of DA19 entered the phosphopentose pathway (PPP) and less entered the glycolytic (EMP) pathway and acetic acid production (Ack-Pta) pathway. Therefore, the differences in activity of key enzyme coincided with increased cell yield based on consumed glucose (Y(X/G)) and decreased production of acetic acid and pyruvate of DA19. G6PDH and ICDH in DH5alpha were up-regulated by addition of adenine, while ACK and PFK were down-regulated. On the other hand, adenine had little effect on those in DA19. The enzymes except PFK in both strains were down-regulated by sodium acetate, especially the activity of ICDH in DH5alpha. These results suggested changed flux of central metabolic pathways were also consistent with the changes of growth properties and byproducts formation.

To study the anti-tumor active constituents from the rhizome of Paris polyphylla var. yunnanensis.The compounds were isolated by column chromatography with silica gel, and purified by Sephadex LH-20 columu chromafography and reverse-phase preparative, HPLC. The structures were identified by spectroscopic methods. The anti-tumor experiment in vitro, the MTT, was used to screen constituents of P. polyphalla var. yunnanensis.Six compounds were obtained and their structures were identified as diosgenin-3-O-alpha-L-arabinofuranosyl (1--4) -beta-D-glycopyranoside (1), pennogenin-3-O-alpha-L-arabino-furanosyl (1--4)-beta-D-glycopyranoside (2), isorhamn etin-3-O-beta-D-glycopyranoside (3), ethyl-alpha-D-fructofuranoside (4), pennogenin-3-O-alpha-L-rhamnopyranosyl (1--4)-[alpha-L-rhamnopyranosyl (1--2)]-beta-D-glycopyranoside (5) and pennogenin-3-O-alpha-L-rhamnopyranosyl (1--4)-alpha-L-rhamnopyranosyl (1--4)-[alpha-L-rhamnopyranosyl (1--2)]-beta-D-glycopyranoside (6).Compounds 1-4 were isolated from this plant for the first time, compounds 3 and 4 was firstly isolated from genus Paris, compound 5 was firstly isolated from the rhizome of this plant. The pharmacological results showed that compounds 1-3, 5 and 6 showed certain inhibition, especially, the activities of compound 5 and 6 were the most significant.

H(2)O(2), plasma membrane H(+)-ATPase (PM H(+)-ATPase) and salicylic acid (SA) play important roles in sensing external stimulation and activating defense responses in plants. However, it remains uncertain whether they are involved and interrelated in response to heat acclimation. Experiments were performed by pharmacological methods, and the relationship and the connection between endogenous H(2)O(2), free SA and PM H(+)-ATPase were investigated in pea plants (Pisum sativum L.) during heat acclimation. The results showed that an accumulation peaks of H(2)O(2), free SA and PM H(+)-ATPase, were detected during heat acclimation at 37 degrees C for 2 h and H(2)O(2) burst appeared before SA accumulation that followed by increase of PM H(+)-ATPase activity (Fig.1). Pretreatments with either scavengers of active oxygen species (dimethyl sulfoxide and ascorbic acid) or antioxidant (reduced glutathione) inhibited the increases in both H(2)O(2) and free SA contents as a part of heat acclimation (Fig.2). Additionally, changes in activity of plasma membrane NADPH oxidase paralleled with H(2)O(2) level during heat acclimation (Figs.1 and 3), implicating that H(2)O(2) might be generated by plasma membrane NADPH oxidase. Moreover, pretreatments with either diphenylene iodonium (DPI), a suicide substrate inhibitor of plasma membrane NADPH oxidase, or dimethylthiourea (DMTU), a quencher of H(2)O(2), could block the increase in free SA content and activity of plasma membrane NADPH oxidase as a part of heat acclimation (Fig.4). According to the assay described above, it is suggested that both H(2)O(2) and PM H(+)-ATPase participate in SA signaling that leads to the development of thermotolerance in pea plant, and H(2)O(2) functions upstream and PM H(+)-ATPase functions downstream of the SA signal. Also, the regulation mechanism of PM H(+)-ATPase activity was investigated, which showed that during heat acclimation, increase of PM H(+)-ATPase activity was independent of PM H(+)-ATPase amount and the enzyme activity may be modulated at post-translational level that may involve in reversible protein phosphorylation (Fig.5).

To evaluate the impact of hypocaloric and hypo-nitrogen parenteral nutrition (PN) on infective complication rate, postoperative hospital stay and treatment cost in postoperative period.120 patients with gastrointestinal tumors with the Nutrition Risk Screening (NRS) score of 3 or 4 undergoing radical gastrectomy in 5 hospitals were randomly assigned into 2 equal groups: control group, receiving PN with 30 (28 - 32) kcal x kg(-1) x d(-1) and nitrogen 0.20 (0.19 - 0.21) g x kg(-1) x d(-1) in regular "3 liter bag", and study group receiving calorie of 18 (range 16 - 20) kcal x kg(-1) x d(-1) and nitrogen of 0.10 (0.09 - 0.11) g x kg(-1) x d(-1) with triple chamber bag. PN support was infused continuously for at least six postoperative days through peripheral vein or peripherally inserted central catheter. The differences between these two groups in blood glucose level, infectious complication, phlebitis, systemic inflammatory response syndrome (SIRS), and duration of hospital stay after operation, and treatment cost. All data were evaluated by both intention to treat (ITT) analysis and per protocol (PP) analysis.There were no significant differences in the clinical baseline and operative types between the two groups. ITT analysis showed that the occurrence of hyperglycemia in postoperative period in the control group was 43.3%, significantly much higher than that in the study group (6.6%, P = 0.000). The infectious complication rate of the study group was 3.3%, significantly lower than that of the control group (16.6%, P = 0.0149), the phlebitis rate of the study group was 0.0%, significantly lower than that of the control group (18.3%, P = 0.0005). The SIRS rate of the study group was 25.0%, significantly lower than that of the control group (45.0%, P = 0.0216). PP analysis showed that the postoperative duration of hospital stay of the control group was 14.1 days +/- 5.8 days, significantly longer than that of the study group (12.4 days +/- 4.0 days, P = 0.047), the total PN cost of the study group was 3411.6 +/- 181.1 Yuan RMB, significantly higher than that of the control group (2945 +/- 162 Yuan RMB, P = 0.000); but the total post-operative cost of treatment of the control group was 13156 +/- 3282 Yuan RMB, significantly higher than that of the study group (11 642 +/- 3019 Yuan RMB, P = 0.010); and the time for compounding of the study group was 5.0 min +/- 1.7 min, significantly shorter than that of the control group (15.4 min +/- 3.7 min, P = 0.000).Hypocaloric and hypo-nitrogen PN in postoperative days 1 - 6 in patients with scores 3 or 4 decreases the rates of hyperglycemia, infectious complications, phlebitis, and SIRS, shortens the postoperative hospital stay, and lowers the cost of treatment comparing with conventional PN. The use of triple chamber bag shortens the compounding time of PN.

Batch cultures of Escherichia coli DH5alpha and DA19 in carbon source limited defined media and continuous cultures in nitrogen source limited defined media were performed to compare the metabolism in both strains. DH5alpha showed improved growth and less acetic acid production after supplementation of adenine, but adenine had little effects on the metabolism of DA19. The proteomic analysis showed that expression of purH was down regulated in DA19 by addition of adenine while was unchanged in DH5alpha, suggesting that the de novo synthesis ability of purine nucleotide in DH5a was poorer than that in DA19. Supplementation of adenine resulted in enhanced expression of gnd and yddS in DH5a, and thus improved the growth.

To evaluate the efficacy, side-effects and quality of life in the advanced colorectal cancer patients treated by irinotecan plus fuorouracil and leucovorin with thalidomide or without thalidomide.Eligible patients were randomly assigned to the treatment group and control group in a 1:1 ratio. In the treatment group, 32 evaluable patients were treated with irinotecan 180 mg/m2 i. v. on day 2, fuorouracil 400 mg/m2 bolus on day 1, 2 at a dose of 1200 mg/m2 civ. for 43 hours; leucovorin 200 mg/m2 i. v. on day 1, 2; thalidomide 300 mg, orally on day 1 - 14, two weeks as a cycle. In the control group, the regimen was the same as in the treatment group except oral intake of thalidomide.The response rate was 28.1% in the treatment group vs. 15.2% in the control group (P = 0.2034) with a median TTP of 3.8 months vs. 2. 5 months (P = 0.1312). Furthermore, there was no statistically difference either between two groups regarding to adverse effects.Irinotecan plus fuorouracil and leucovorin without oral intake of thalidomide is as effective and tolerable as irinotecan plus fuorouracil and leucovorin combined with oral thalidomide for advanced colorectal cancer.

To determine whether cysteinyl leukotriene receptors (CysLT1 and CysLT2) are upregulated in acute neuronal injury after focal cerebral ischemia in mice, and to confirm CysLT1 receptor localization.After permanent focal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO), neurological deficits and neuron loss were determined at various time points within 48 h. The mRNA expressions of CysLT1 and CysLT2 receptors in the brain were analyzed by RT-PCR. CysLT1 receptor localization was detected by double immunofluorescence.Neurological deficits and neuron loss were found 6, 24 and 48 h after MCAO. The mRNA expressions of both CysLT1 and CysLT2 receptors were upregulated in the ischemic hemisphere 1, 24, and 48 h after MCAO with peaks at 24 h. The CysLT1 receptor was selectively localized in neurons 24 h after MCAO.CysLT1 and CysLT2 receptors are upregulated in acute neuronal injury after focal cerebral ischemia, and the CysLT1 receptor is localized in neurons after ischemia.

Phytochemical investigation of chemical constituents of the CHCl3 fraction of 75% alcohol extract from the leaves of Cyclocarya paliurus has resulted in the isolation of five compounds, alpha-boswellic (I), beta-boswellic (II), Oleanolic acid (III), 4-hydroxyl-3-methoxyl-benzoic acid (IV) and 3,6,3',5'-tetramenthox-5,7,4'-trihydroxy-flavonol (V). Their structures were elucidated on the basis of spectroscopic analyses, especially NMR. Compunds I, II, IV and V were obtained from this plant for the first time.

The application of the reverse engineering and rapid prototyping technologies in the design of respiratory masks is introduced in this paper. Practice indicates that the technologies can reduce the cost and save the time in product developments.

In this study, we have identified 876 polymorphism sites in 145 complete or partial genomes of SARS-CoV available in the NCBI GenBank. One hundred and seventy-four of these sites existed in two or more SARS-CoV genome sequences. According to the sequence polymorphism, all SARS-CoVs can be divided into three groups: (I) group 1, animal-origin viruses (such as SARS-CoV SZ1, SZ3, SZ13 and SZ16); (II) group 2, all viruses with clinical origin during first epidemic; and (III) group 3, SARS-CoV GD03T0013. According to 10 special loci, group 2 again can be divided into genotypes C and T, which can be further divided into sub-genotypes C1–C4 and T1–T4. Positive Darwinian selections were identified between any pair of these three groups. Genotype C gives neutral selection. Genotype T, however, shows negative selection. By comparing the death rates of SARS patients in the different regions, it was found that the death rate caused by the viruses of the genotype C was lower than that of the genotype T. SARS-CoVs might originate from an unknown ancestor.

In wireless sensor networks (WSNs), sensor location plays a critical role in many applications. Aiming at that all the nodes in sensor networks need to be located, the paper presents an assisting localization method which locates the sensor nodes with respect to the outside reference signal sources and can achieve efficient location discovery without using beacons. We give the localization models based on TDOA, TOA and AOA. Despite the measurement errors, the method provides sufficient location information and accuracy to support basic network functions. Furthermore the method based on AOA estimation model does not increase the complexity or cost of construction of the sensor nodes. Extensive simulation experiment results show that the proposed method can locate the sensor nodes effectively

To determine the expressions of cysteinyl leukotriene receptors, CysLT1 and CysLT2, in airway eosinophilic inflammation of OVA-induced asthmatic mice and the modulation by montelukast, a CysLT1 receptor antagonist.Asthma model was induced by chronic exposure to ovalbumin (OVA) in C57BL/6 mice. The eosinophils in bronchoalveolar lavage (BAL) fluid and lung tissues were counted, IL-5 level in BAL fluid was measured, and CysLT1 and CysLT2 receptor mRNA expressions were detected by semi-quantitative RT-PCR.Montelukast (6 mg/kg, once per day for 20 d) significantly suppressed the increased eosinophils in BAL fluid and lung tissue, and increased IL-5 level in BAL fluid in OVA challenged mice. OVA challenge increased CysLT1 but decreased CysLT2 receptor mRNA expression. Montelukast inhibited the increased CysLT1 but not the reduced CysLT2 expression after OVA challenge.CysLT receptors are modulated immunologically, and montelukast inhibits up-regulation of CysLT1 receptor and airway eosinophilic inflammation in asthmatic mice.

SARS-CoV is the causative agent of severe acute respiratory syndrome (SARS) which has been associated with outbreaks of SARS in Guangdong, Hong Kong and Beijing of China, and other regions worldwide. SARS-CoV from human has shown some variations but its origin is still unknown. The genotyping and phylogeny of SARS-CoV were analyzed and reported in this paper.Full or partial genomes of 44 SARS-CoV strains were collected from GenBank. The genotype, single nucleotide polymorphism and phylogeny of these SARS-CoV strains were analyzed by molecular biological, bioinformatic and epidemiological methods.There were 188 point mutations in the 33 virus full genomes with the counts of mutation mounting to 297. Further analysis was carried out among 36 of 188 loci with more than two times of mutation. All the 36 mutation loci occurred in coding sequences and 22 loci were non-synonymous. The gene mutation rates of replicase 1AB, S2 domain of spike glycoprotein and nucleocapsid protein were lower (0.079% - 0.103%). There were 4 mutation loci in S1 domain of spike glycoprotein. The gene mutation rate of ORF10 was the highest (3.333%) with 4 mutation loci in this small domain (120 bp) and 3 of 4 loci related to deletion mutation. By bioinformatics processing and analysis, the nucleotides at 7 loci of genome (T:T:A:G:T:C:T/C:G:G:A:C:T:C) can classify SARS-CoV into two types. Therefore a novel definition is put forward that according to these 7 loci of mutation, 40 strains of SARS-CoV in GenBank can be grouped into two genotypes, T:T:A:G:T:C:T and C:G:G:A:C:T:C, and named as SARS-CoV Yexin genotype and Xiaohong genotype. The two genotypes can be further divided into some sub-genotypes. These genotypes can also be approved by phylogenetic tree of three levels of 44 loci of mutation, spike glycoprotein gene and complete genome sequence. Compared to various strains among SARS-CoV Yexin genotype and Xiaohong genotype, GD01 strain of Yexin genotype is more closely related to SARS-CoV like-virus from animals.The results mentioned above suggest that SARS-CoV is responding to host immunological pressures and experiencing variation which provide clues, information and evidence of molecular biology for the clinical pathology, vaccine developing and epidemic investigation.

AIM: To construct a recombined human AFP eukaryotic expression vector for the purpose of gene therapy and target therapy of hepatocellular carcinoma (HCC). METHODS: The full length AFP-cDNA of prokaryotic vector was digested, and subcloned to the multi-clony sites of the eukaryotic vector. The constructed vector was confirmed by enzymes digestion and electrophoresis, and the product expressed was detected by electrochemiluminescence and immunofluorescence methods. RESULTS: The full length AFP-cDNA successfully cloned to the eukaryotic vector through electrophoresis, 0.9723 IU/mL AFP antigen was detected in the supernatant of AFP-CHO by electrochemiluminescence method. Compared with the control groups, the differences were significant (P < 0.05). AFP antigen molecule was observed in the plasma of AFP-CHO by immunofluorescence staining. CONCLUSION: The recombined human AFP eukaryotic expression vector can express in CHO cell line. It provides experimental data for gene therapy and target therapy of hepatocellular carcinoma.

p28, a 28kD protein from toad (Bufo bufo gargarizans) oocytes, was identified by using p13 suc1 -agaroseaffinity chromatography. Sequence homology analysis of the full-length cDNA of p28(Gene Bank accessionnumber: AF 314091) indicated that it encodes a protein containing 224 amino-acids with about 55% iden-tities and more than 70% positives to human, rat or mouse UCH-L1, and contains homological functionaldomains of UCH family. Anti-p28 monoclonal antibody, on injecting into the oocytes, could inhibit theprogesterone-induced resumption of meiotic division in a dose-dependent manner. The recombinant pro-tein p28 showed similar SDS/PAGE behaviors to the native one, and promoted ubiquitin ethyl ester hydrolysis,a classical catalytic reaction for ubiquitin carboxyl terminal hydrolases (UCHs). The results in this paperreveal that a novel protein, p28, exists in the toad oocytes, is a UCH L1 homolog, was engaged in the processof progesterone-induced oocyte maturation possibly through an involvement in protein turnover anddegradation.Key words: p28, cDNA clone, recombinant expression, ubiquitin carboxyl terminal hydrolase, oocytematuration.

Background & Aims: In this study, we investigated the role of leptin and its receptors (Ob-R) in profibrogenic responses in the liver using Zucker (fa/fa) rats, a natural occurring Ob-R–deficient animal. Methods: Male Zucker (fa/fa) rats and their lean (+/?) littermates were given intraperitoneal injections of thioacetamide (TAA) (200 mg/kg body wt, 3 times/wk) for 4–8 weeks, and progression of hepatic fibrosis was evaluated. In vitro transactivation of hepatic stellate cells (HSCs) isolated from Zucker rats was evaluated by Western blotting and immunocytochemistry for α-smooth muscle actin and type I collagen. Further, a long-form Ob-R (Ob-Rb) in sinusoidal endothelial cells (SECs) and Kupffer cells was identified by reverse-transcription polymerase chain reaction. Moreover, transforming growth factor (TGF)-β1 messenger RNA in LSE cells, a human SEC-derived cell line, was measured by Northern blotting. Results: Although the normal liver does not produce leptin, activated HSCs produced leptin in vivo during fibrogenesis caused by TAA. In Zucker rats, TAA-induced hepatic fibrosis was prevented almost completely, whereas induction of TGF-β1 and activation of HSCs were abolished. It is less likely, however, that leptin plays an essential role in the activation of HSCs as a strong autocrine regulator, because HSCs isolated from Zucker rats undergo normal transactivation process in vitro. In contrast, SECs and Kupffer cells contain Ob-Rb, through which leptin up-regulates the expression of matrix remodeling genes including TGF-β1. Conclusions: Collectively, these findings indicated that leptin and its functional receptors (Ob-Rb) play a pivotal role in profibrogenic responses in the liver. GASTROENTEROLOGY 2002;122:1399-1410

113 Background: PIK3CAgene mutations are the most common activating mutations in human breast cancer (BC). Its association with the resistance to herceptin in HER2-positive BC was reported, but whether it’s related to the efficacy of neoadjuvant chemotherapy is unknown. Methods: We reviewed records of 108 patients with BC treated with epirubicin plus docetaxel neoadjuvant chemotherapy between June 2005 and April 2011. 92 patients’ clinical and pathologic objective response data were collected. FFPE tumor biopsies obtained at the time of diagnoses (and before treatment) were collected. EGFR, KRAS, BRAF, PIK3CA mutations and HER2, PTEN, EGFR mRNA expression were analyzed by liquidchip technology. Chi-square test and Fisher’s exact test were used in statistical analysis. A p value less than 0.05 was considered statistically significant. Results: No mutation was detected in EGFR, KRAS and BRAF. The mutation rate of PIK3CA was 35.1% (38/108), and mutation rates of E542K, E545K, H1047L and H1047R were 3.7, 10.2, 3.7 and 17.6% respectively. More PIK3CA mutations were detected in HER2 high expression tumors (p=0.018). No correlation was found between PIK3CA mutations and PTEN, EGFR expression. Clinical response (CR+PR, 63/92) was associated with PIK3CA mutations (p=0.002). However, PIK3CA mutations rate between pathologic complete release (pCR, 7/92) and non-pCR group was not significantly different (p=0.422). Conclusions: PIK3CA mutation may predict better clinical response in BC patients treated with epirubicin plus docetaxel neoadjuvant chemotherapy. More studies are needed to validate these results and the correlation between PIK3CA mutations and pCR.