Lonicera flos and Cnicus japonicus extracts improved egg quality partly by modulating antioxidant status, inflammatory-related cytokines and shell matrix protein expression of oviduct in laying hens.

This study was conducted to investigate the effects of Lonicera flos and Cnicus japonicus extracts (LCE) on the laying performance, egg quality, morphology, antioxidant status, inflammatory-related cytokines, and shell matrix protein expression of oviduct in laying hens. A total of 1,728 Roman Pink laying hens aged 73-wk-old were randomly assigned into 4 groups (18 replicates/group, 24 layers/replicate) fed basal diets supplemented with 0, 300, 500, and 1,000 mg of LCE per kg of diet, respectively. The trial lasted for 11 wk, including 2-wk adjustment period and 9-wk testing period. The results indicated that laying hens fed diets supplemented with LCE linearly increased egg weight, yolk color and shell thickness at wk 78 and albumen height, Haugh unit and shell thickness at wk 83 (P < 0.05). At wk 78, LCE groups linearly affected the hydrogen peroxide content in magnum (P < 0.05) and 300 mg/kg LCE groups had the highest catalase activity in isthmus (P < 0.05). At wk 83, LCE groups linearly reduced (P < 0.05) hydrogen peroxide content in the magnum and isthmus and malondialdehyde content in the uterus whereas increased catalase activity in isthmus (P < 0.05). Furthermore, LCE levels quadratically affected glutathione peroxidase activity in isthmus at wk 83 (P < 0.05). At wk 78, the mRNA expressions of inducible nitric oxide synthase and interferon-γ in isthmus and ovalbumin and ovocleidin-116 in uterus had linear effects in response to LCE levels (P < 0.05) and 1,000 mg/kg LCE group had the lowest mRNA expression of interleukin-6 in magnum (P < 0.05). At wk 83, LCE supplementation linearly decreased the mRNA expression of interleukin-1ß, interferon-γ and tumor necrosis factor-α in magnum and tumor necrosis factor-α and inducible nitric oxide synthase in uterus (P < 0.05). It is concluded that LCE improved egg quality partly by modulating antioxidant status, inflammatory-related cytokines and shell matrix protein expression of oviduct in laying hens.

Antibiofilm activity against Staphylococcus aureus and content analysis of Taraxacum Officinale phenolic extract.

Taraxacum Officinale, commonly called dandelion, is herbaceous perennial belonging to the family of Asteraceae, having good antibacterial effects which are related to its phenolic substances. In this study, the effect of phenolic contents as well as the antibiofilm activity against Staphylo- coccus aureus of phenolic extract from T. Officinale were evaluated in vitro. With 70% metha- nol-water (v/v) as a solvent, the dandelion was extracted by ultrasonic assisted extraction method. Subsequent identification and quantification of phenol in extract was carried out using High Performance Liquid Chromatography (HPLC). The minimum inhibitory concentration and anti- bacterial kinetic curve of dandelion phenolic extract were analyzed by spectrophotometry. Changes in extracellular alkaline phosphatase (AKP) contents, electrical conductivity, intracellular protein contents, and DNA of S. aureus after the action of dandelion phenolic extract were determined to study its effect on the permeability of S. aureus cell wall and cell membrane. The results showed that chlorogenic acid (1.34 mg/g) was present in higher concentration, followed by lute- olin (1.08 mg/g), ferulic acid (0.22 mg/g), caffeic acid (0.21 mg/g), and rutin (0.19 mg/g) in the dandelion phenolic extract. The minimum inhibitory concentration (MIC) of dandelion phenolic extract against S. aureus was 12.5 mg/mL. The antibacterial kinetic curve analysis showed that the inhibitory effect of dandelion phenolic extract on S. aureus was mainly in the exponential growth phase. After applying the dandelion phenolic extract, the growth of S. aureus was signifi- cantly inhibited entering into the decay phase early. Furthermore, after the action of dandelion, the extracellular AKP contents of S. aureus, the electrical conductivity and the extracellular protein contents were all increased. The phenolic extract also affected the normal reproduction of S. aureus. These results suggest that dandelion has an inhibitory effect on S. aureus, and the mechanism of its action was to destroy the integrity of the cell walls and cell membranes.

[Prognostic study of 229 follicular lymphoma patients treated with rituximab combined with chemotherapy].

Objective: To explore the clinical characteristics of follicular lymphoma (FL) in the era of rituximab combined with chemotherapy and the prognostic significance of the follicular lymphoma international prognostic index (FLIPI), follicular lymphoma international prognostic index 2 (FLIPI2), international prognostic index (IPI), revised international prognostic index (R-IPI), National Comprehensive Cancer Network international prognostic index (NCCN-IPI) among Chinese patients. Methods: 229 FL patients who were treated initially with rituximab combined with CHOP-like (cyclophosphamide, doxorubicin, vincristine and prednisone) chemotherapy from November 2008 to April 2018 were analyzed retrospectively and all were scored by the above clinical index. Univariate and multivariate survival analysis were performed on 201 patients who completed the treatment and were followed regularly. Results: In the univariate survival analysis, age>60 years, hemoglobin<120 g/L, elevated serumß(2)- macroglobulin, involvement of bone marrow and elevated CRP were the risk prognostic factors for overall survival (OS) and progression free survival (PFS). Moreover, the analysis of the OS and PFS between rituximab (R) maintenance (RM) group and non-maintenance (non-RM) group showed that the OS and PFS of RM group were better than those of non-RM. In the multivariate analysis of OS, hemoglobin<120 g/L, involvement of bone marrow, elevated CRP and non-RM were independent prognostic factors. In the multivariate analysis of PFS, hemoglobin<120 g/L, CRP and non-RM were independent prognostic factors. When FLIPI2 was included in the multivariate analysis, CRP and FLIPI2 were independent prognostic factors in both OS and PFS, and non-RM was independent prognostic factors in PFS. Conclusion: FLIPI2 is the better risk stratification in FL patients in the era of rituximab.

[Prognostic significance of NCCN-International Prognostic Index (NCCN-IPI) for patients with peripheral T-cell lymphoma treated with CHOP-based chemotherapy].

Objective: To validate the prognostic value of NCCN-International Prognostic Index (NCCN-IPI) for patients with peripheral T-cell lymphoma (PTCL) treated with CHOP-based chemotherapy. Methods: A retrospective analysis in 162 PTCL patients who were initially diagnosed and treated in Rui Jin Hospital from January 2003 to May 2013 was conducted. Baseline characteristics were collected, and survival analysis was performed according to the IPI and NCCN-IPI model. Results: The estimated 5-year overall survival (OS) rate and progression free survival (PFS) rate were 33% and 20%, with median OS and PFS of 17.0 months and 9.2 months, respectively. Multivariate analysis indicated ECOG score (PFS: HR=2.418, 95%CI 1.535-3.809, P<0.001; OS: HR=2.347, 95%CI 1.435-3.839, P= 0.001) , specific extra-nodal sites (PFS: HR=1.800, 95%CI 1.216-2.665, P=0.003; OS: HR=1.608, 95% CI 1.054-2.454, P=0.027) and pathology type (PFS: HR=0.424, 95% CI 0.184-0.975, P=0.043; OS: HR=0.276, 95% CI 0.087-0.877, P=0.029) were independent prognostic factors of OS and PFS for the patients with PTCL. The survival rates of low risk patients based on NCCI-IPI were remarkably higher than the counterparts based on IPI (5-year OS 74% vs 54%, χ(2)=5.041, P=0.025, 5-year PFS 50% vs 38%, χ(2)= 5.295, P=0.021) . NCCN-IPI was outstanding to identify the subgroup of low risk patients with PTCL, who may benefit from conventional chemotherapy such as CHOP or CHOP-like regimen. Conclusion: NCCN-IPI is more powerful for low risk PTCL patients and a strong supplement for IPI.

Therapeutic efficacy of atypical antipsychotic drugs by targeting multiple stress-related metabolic pathways.

Schizophrenia (SZ) is considered to be a multifactorial brain disorder with defects involving many biochemical pathways. Patients with SZ show variable responses to current pharmacological treatments of SZ because of the heterogeneity of this disorder. Stress has a significant role in the pathophysiological pathways and therapeutic responses of SZ. Atypical antipsychotic drugs (AAPDs) can modulate the stress response of the hypothalamic-pituitary-adrenal (HPA) axis and exert therapeutic effects on stress by targeting the prefrontal cortex (PFC) and hippocampus. To evaluate the effects of AAPDs (such as clozapine, risperidone and aripiprazole) on stress, we compared neurochemical profile variations in the PFC and hippocampus between rat models of chronic unpredictable mild stress (CUMS) for HPA axis activation and of long-term dexamethasone exposure (LTDE) for HPA axis inhibition, using an ultraperformance liquid chromatography-mass spectrometry (UPLC-MS/MS)-based metabolomic approach and a multicriteria assessment. We identified a number of stress-induced biomarkers comprising creatine, choline, inosine, hypoxanthine, uric acid, allantoic acid, lysophosphatidylcholines (LysoPCs), phosphatidylethanolamines (PEs), corticosterone and progesterone. Specifically, pathway enrichment and correlation analyses suggested that stress induces oxidative damage by disturbing the creatine-phosphocreatine circuit and purine pathway, leading to excessive membrane breakdown. Moreover, our data suggested that the AAPDs tested partially restore stress-induced deficits by increasing the levels of creatine, progesterone and PEs. Thus, the present findings provide a theoretical basis for the hypothesis that a combined therapy using adenosine triphosphate fuel, antioxidants and omega-3 fatty acids as supplements may have synergistic effects on the therapeutic outcome following AAPD treatment.

Physiological Responses of Aphanizomenon flos-aquae Under the Stress of Sagittaria sagittifolia Extract.

The algal growth and physiological characters of Aphanizomenon flos-aquae were studied under the stress of Sagittaria sagittifolia extract. The results showed that the growth of A. flos-aquae was significantly inhibited by S. sagittifolia extract. The exopolysaccharide (EPS), total soluble protein, intracellular phosphorus (o-PO4-P) contents and malondialdehyde (MDA) contents in A. flos-aquae cells increased significantly. These results suggested that A. flos-aquae can adapt to stress by increasing its normal metabolic activity. The algal cellular antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), were triggered to different degrees when exposed to S. sagittifolia extract. The MDA contents and activities of SOD, CAT and POD in algal cells suggested that oxidative damage induced by S. sagittifolia extract via the oxidation of ROS (O2·-) might be an important factor responsible for the inhibition of the growth of A. flos-aquae. In addition, SOD may be an important site for the inhibition of S. sagittifolia extract on A. flos-aquae cells. These results indicate that S. sagittifolia may be a good candidate for controlling A. flos-aquae blooms.

[Efficacy of additional two cycles of rituximab administration for patients with diffuse large B-cell lymphoma in first remission].

Objective: To analyze the efficacy of additional two cycles of rituximab administration for Chinese patients with diffuse large B-cell lymphoma (DLBCL) in first complete remission (CR) after six cycles of standard 21-day rituximab plus cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP21). Methods: Retrospective analysis was performed in 351 patients with DLBCL diagnosed from March 2003 to March 2012. International Prognosis Index (IPI), Revised (R)-IPI and National Comprehensive Cancer Network (NCCN)-IPI were calculated for each patient. Patients were divided into GCB and non-GCB subtype according to Han's Classification. Progression-free survival (PFS) and overall survival (OS) were analyzed using Kaplan-Meier methods. Results: 282 (80.3%) patients achieved CR and 132 (46.8%) of 282 cases received additional two rituximab therapy. The other 150 (53.2%) patients entered into observation on the intention of the patients. No significant difference was observed in baseline characteristics between the two groups. 3-year estimated PFS for additional rituximab group and observation group were 80.0% and 78.1% (P=0.334), while 3-year estimated OS were 89.7% vs. 86.1% (P=0.452). By subgroup analysis, prolonged PFS were observed in R-IPI low-risk and NCCN-IPI low-risk patients after additional two rituximab cycles. Conclusion: For patients with DLBCL in first remission after standard six cycles of R-CHOP21 regimen, additional two cycles of rituximab maintenance did not significantly improve the general prognosis, but low-risk subgroups of R-IPI and NCCN-IPI could benefit from this regimen.

Pyrazinamide resistance in Mycobacterium tuberculosis arises after rifampicin and fluoroquinolone resistance.

BACKGROUND: Multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains of Mycobacterium tuberculosis (TB) constitute a major public health concern. OBJECTIVE: To determine the timing of pncA mutations that confer pyrazinamide (PZA) resistance in relation to mutations conferring resistance to isoniazid (INH) and rifampicin (RMP). DESIGN: Isolates from two major urban centres--Paris (101 strains) and Shanghai (171 strains)--were investigated for the association of pncA mutations with resistance to drugs other than PZA. RESULTS: The proportion of pncA mutations found in INH-monoresistant strains was not increased. CONCLUSION: pncA mutations associated with PZA resistance were found almost exclusively in MDR-TB strains, underlining the importance of determining PZA resistance when treating MDR- or XDR-TB.

Microstructure assessment of the thalamus in Wilson's disease using diffusion tensor imaging.

AIM: To assess diffusion changes of the thalamus in Wilson's disease using diffusion tensor imaging (DTI). MATERIALS AND METHODS: Fifteen patients with Wilson's disease and an abnormal signal in the thalamus (designated as group 1) and 18 patients with Wilson's disease with a normal-appearing thalamus (designated as group 2) at conventional magnetic resonance imaging (MRI) were recruited. Fifteen age-matched and sex-matched healthy volunteers were also enrolled as the control group (designated as group 3). The fractional anisotropy (FA), primary eigenvalue (λ1), second eigenvalue (λ2), and third eigenvalue (λ3) of the thalamus were measured and the differences were compared. RESULTS: The FA values of the thalamus were different in the three groups (group 1: 0.36 ± 0.02; group 2: 0.38 ± 0.02; group 3: 0.43 ± 0.02; F = 54.51, p < 0.001). A statistically significant difference was observed between group 1 and group 2 (p = 0.003), group 1 and group 3 (p = 0.001), and group 2 and group 3 (p < 0.001). The λ1, λ2, and λ3 values of the thalamus were different in the three groups (1.11 ± 0.06 mm(2)/s, 1.11 ± 0.06 mm(2)/s, and 1.10 ± 0.04 mm(2)/s of λ1 in group 1, group 2, and group 3, respectively; 0.82 ± 0.08 mm(2)/s, 0.78 ± 0.05 mm(2)/s, and 0.72 ± 0.02 mm(2)/s of λ2 in group 1, group 2, and group 3, respectively; 0.52 ± 0.05 mm(2)/s, 0.49 ± 0.06 mm(2)/s, and 0.42 ± 0.06 mm(2)/s of λ3 in group 1, group 2, and group 3, respectively; F = 1.65, p = 0.203 of λ1; F = 10.55, p < 0.001 of λ2; F = 4.21, p = 0.021 of λ3; respectively). A statistically significant difference in the λ2 value was observed between group 1 and group 3 (p < 0.001) and group 2 and group 3 (p = 0.005). A statistically significant difference in the λ3 value was also observed between group 1 and group 3 (p = 0.007). No significant difference in the λ1 value was noted between each of the two groups. CONCLUSIONS: Damage of the thalamus in Wilson's disease patients can be detected using DTI. DTI may provide information regarding thalamus damage in patients with Wilson's disease before abnormal signals on conventional MRI.

High levels of vitamin E affect retinol binding protein but not CYP26A1 in liver and hepatocytes from laying hens.

This study was conducted to determine the influence of vitamin E on the retinol binding protein (RBP) and cytochrome p450 family 26 subfamily A polypeptide 1 (CYP26A1), which are specific transporters and catabolic enzymes of vitamin A, respectively. In the in vivo experiments, a total of 450 laying hens was fed 5 levels of vitamin E (0, 20, 80, 320, and 1,280 IU/kg of feed) supplementation. For the in vitro assays, hepatocytes from laying hens were cultured in 4 levels of α-tocopherol (0, 10, 50, and 100 µM). High dietary vitamin E increased the concentration of vitamin A in liver (P < 0.05). The RBP and its mRNA expression in liver and hepatocytes were markedly inhibited by dietary vitamin E (320 and 1,280 IU/kg) and α-tocopherol (100 µM) in culture medium (P < 0.05). However, CYP26A1 and its mRNA expression were not affected by vitamin E in both liver and hepatocytes (P > 0.05). The results indicate that excessive vitamin E could increase the concentration of vitamin A in liver by inhibiting RBP synthesis in hepatocytes.

Wnt5a promotes differentiation of human dental papilla cells.

AIM: To investigate the role of Wnt5a in the process of differentiation of human dental papilla cells (HDPCs). METHODOLOGY: Recombinant adenovirus encoding full-length Wnt5a cDNA was constructed to investigate the biological role of Wnt5a on the differentiation of HDPCs. The effect of Wnt5a on HDPCs differentiation was determined by ALP activity assay, ALP staining and mineral induction assay. Mineralization-related gene expressions were assessed by RT-PCR. RESULTS: Immunostaining revealed Wnt5a expression in the odontoblast layer and dental papilla tissue. Over-expression of Wnt5a by transfecting HDPCs with an Wnt5a-carrying construct increased ALPase activity and the formation of mineralized nodules of HDPCs. RT-PCR analysis showed that the expressions of mineralization-related genes, such as bone sialoprotein, collagen type I, osteonectin, osteopontin (OCN), dentine matrix protein-1 were up-regulated by Wnt5a. CONCLUSIONS: Wnt5a promoted differentiation of HDPCs.

Inclusion phenomena of clove oil with alpha-, beta-, gamma- and heptakis (2,6-di-O-methyl)-beta-cyclodextrin.

Inclusion interactions of alpha-, beta-, gamma- and heptakis (2,6-di-O-methyl)-beta-cyclodextrin (DMbeta-CD) as hosts with clove oil (an impure eugenol, I-Eug) as guest in aqueous solution were investigated by fluorescence emission spectra. The binding constants of different hosts to I-Eug in aqueous solution decreased in the order: gamma- > beta- > DMbeta- > alpha-CD. Two solid supramolecular inclusion complexes, I-Eug-beta-CD and I-Eug-gamma-CD, were prepared and characterised by nuclear magnetic resonance, powder X-ray diffraction, infrared spectroscopy, and thermogravimetric analysis. All the results proved the formation of I-Eug-CD. The inclusion differences between I-Eug and pure eugenol were discussed. The relative contents of the main component eugenol (Eug), second component (eugenol acetate, Eua) and others in I-Eug were found to be fairly different before and after being included by beta-CD, according to the data obtained from high performance liquid chromatography. This could be a practical method to extract the effective components (Eug and Eua) from I-Eug.

The effect of herbal medicine baicalin on pharmacokinetics of rosuvastatin, substrate of organic anion-transporting polypeptide 1B1.

The aim of this study was to explore potential herb-drug interaction between baicalin and rosuvastatin, a typical substrate for organic anion-transporting polypeptide 1B1 (OATP1B1) related to different OATP1B1 haplotype groups. Eighteen unrelated healthy volunteers who were CYP2C9*1/*1 with different OATP1B1 haplotypes (six OATP1B1*1b/*1b, six OATP1B1*1b/*15, and six OATP1B1*15/*15) were selected to participate in this study. Rosuvastatin (20 mg orally) pharmacokinetics after coadministration of placebo and 50-mg baicalin tablets (three times daily orally for 14 days) were measured for up to 72 h by liquid chromatography-mass spectrometry in a two-phase randomized crossover study. After baicalin treatment, the area under the plasma concentration-time curve (AUC)(0-72) and AUC(0-infinity) of rosuvastatin decreased by 47.0+/-11.0% (P=0.001) and 41.9+/-7.19% (P=0.001) in OATP1B1*1b/*1b, 21.0+/-20.6% (P=0.035) and 23.9+/-8.66% (P=0.004) in OATP1B1*1b/*15, and 9.20+/-11.6% (P=0.077) and 1.76+/-4.89% (P=0.36) in OATP1B1*15/*15, respectively. Moreover, decreases of both AUC(0-72) and AUC(0-infinity) of rosuvastatin among different haplotype groups were significantly different (P=0.002 and <0.001). Baicalin reduces plasma concentrations of rosuvastatin in an OATP1B1 haplotype-dependent manner.

Efficacy of hyperbaric oxygen on survival of random pattern skin flap in diabetic rats.

OBJECTIVES: This study was designed to determine if hyperbaric oxygen improved random pattern skin flap survival in diabetic rats. METHODS: Cranially-based, 4 x 10 cm dorsal skin flaps were raised in 38 diabetic rats induced by streptozocin (STZ). The animals were randomly divided equally into two groups. Group A was a control group observed in the room air and Group B was the experimental group, which received hyperbaric oxygen (HBO2) therapy. The HBO2 regimen consisted of 90 minutes of treatment with 100% O2 at 2.5 ATA (atmosphere absolute ATA) per day for 7 consecutive days. On the 7th postoperative day, we measured the necrotic flap area and the new growth number of capillary vessel and the granulation tissue thickness. RESULTS: The percentage of necrosis flap area for group A was 50.5 +/- 10.5%; for group B it was 38.5 +/- 9.3%. The reduction in necrosis flap area was highly significant (p < 0.01) compared with controls. Also, new-growth capillary vessel and granulation tissue thickness were statistically different between the two groups. CONCLUSIONS: The findings of this study demonstrated beneficial effects of HBO2 in improving diabetic rat dorsal skin flap survival.

Tetramethylpyrazine production from glucose by a newly isolated Bacillus mutant.

2,3,5,6-Tetramethylpyrazine (TTMP) was produced using a newly isolated Bacillus mutant. Culture medium optimization studies showed that soytone, an enzyme-hydrolysate of soybean meal, with the supplementation of vitamins, can fully replace yeast extract plus peptone in supporting TTMP production from glucose. In a 5-l fermenter, using the optimized medium which contained 20% glucose, 5% soytone, 3% (NH(4))(2)HPO(4), and vitamin supplements, fermentations were carried out with stirring at 700 rpm, air flow at 1.0 vvm, controlled pH at 7.0, and temperature at 37 degrees C. TTMP reached 4.33 g l(-1) after 64.6 h cultivation. A product recovery method was described, which involved evaporation, crystallization, and lyophilization. The product purity was 99.88%, determined by GC with the normalization method. The main impurities were 2,3,5-trimethylpyrazine (0.09%) and 2-ethyl-3,5,6-trimethylpyrazine (0.02%), which were identified by GC/MS. (13)C NMR determination also gave a consistent result. Natural and high purity of the product and the utilization of cheap green renewable materials make this process promising to compete with TTMP chemical synthetic methods.

In vivo modification of Na(+),K(+)-ATPase activity in Drosophila.

We have constructed and characterized transgenic Drosophila lines with modified Na(+),K(+)-ATPase activity. Using a temperature dependent promoter from the hsp70 gene to drive expression of wild-type alpha subunit cDNA, we can conditionally rescue bang-sensitive paralysis and ouabain sensitivity of a Drosophila Na(+),K(+)-ATPase alpha subunit hypomorphic mutant, 2206. In contrast, a mutant alpha subunit (alpha(D369N)) leads to increased bang-sensitive paralysis and ouabain sensitivity. We can also generate temperature dependent phenotypes in wild-type Drosophila using the same hsp70 controlled alpha transgenes. Ouabain sensitivity was as expected, however, both bang sensitive paralysis or locomotor phenotypes became more severe regardless of the type of alpha subunit transgene. Using the Gal4-UAS system we have limited expression of alpha transgenes to cell types that normally express a particular Drosophila Na(+),K(+)-ATPase beta (Nervana) subunit isoform (Nrv1 or 2). The Nrv1-Gal4 driver results in lethality while the Nrv2-Gal4 driver shows reduced viability, locomotor function and uncontrolled wing beating. These transgenic lines will be useful for disrupting function in a broad range of cell types.

[Identification of medicinal plant Xanthium L. produced in China].

OBJECTIVE: To gain a clear idea on the resources and pharmacognostic identification of medicinal plant Xanthium in China. METHOD: Identification of botanical origin, analysis of fruit shapes and properties, microscopic characteristics, TLC and UV. RESULT: Identification criteria have been worked out for Xanthium and its confused species. CONCLUSION: The resources of medicinal plant Xanthium may be appropriately expanded.

Characterization of Drosophila insulin receptor substrate.

Insulin receptor substrate (IRS) proteins are phosphorylated by multiple tyrosine kinases, including the insulin receptor. Phosphorylated IRS proteins bind to SH2 domain-containing proteins, thereby triggering downstream signaling pathways. The Drosophila insulin receptor (dIR) C-terminal extension contains potential binding sites for signaling molecules, suggesting that dIR might not require an IRS protein to accomplish its signaling functions. However, we obtained a cDNA encoding Drosophila IRS (dIRS), and we demonstrated expression of dIRS in a Drosophila cell line. Like mammalian IRS proteins, the N-terminal portion of dIRS contains a pleckstrin homology domain and a phosphotyrosine binding domain that binds to phosphotyrosine residues in both human and Drosophila insulin receptors. When coexpressed with dIRS in COS-7 cells, a chimeric receptor (the extracellular domain of human IR fused to the cytoplasmic domain of dIR) mediated insulin-stimulated tyrosine phosphorylation of dIRS. Mutating the juxtamembrane NPXY motif markedly reduced the ability of the receptor to phosphorylate dIRS. In contrast, the NPXY motifs in the C-terminal extension of dIR were required for stable association with dIRS. Coimmunoprecipitation experiments demonstrated insulin-dependent binding of dIRS to phosphatidylinositol 3-kinase and SHP2. However, we did not detect interactions with Grb2, SHC, or phospholipase C-gamma. Taken together with published genetic studies, these biochemical data support the hypothesis that dIRS functions directly downstream from the insulin receptor in Drosophila.

[Determination of tetrandrine in human plasma by high-performance liquid chromatography].

The paper reports the determination of tetrandrine by high performance liquid chromatography (HPLC). A C18 column was used and the mobile phase was methanol-water(80:20) containing 0.03% triethylamine, detective wave was at 282 nm, the internal standard was diazepam, the sample was extracted with ether. The linear range was from 0.289 microgram.ml-1 to 4.618 micrograms.ml-1, the average recovery rate was 95.8%. The coefficients of variation of within-day and day-to-day were less than 5%. This method is simple, rapid and accurate. It can be used in biomedical sample analysis of tetrandrine.

[A study of quality standards for fuyanke granule].

OBJECTIVE: To establish the quality standards for fuyanke(FYK) granule. METHODS: The phellodendron Chinese schneid, polygonum cuspidatum, sieb. et Zucc and glycyrrihza uralensis fisch in FYK granule were identified by thin-layer chromatography(TLC). The contents of berberine were determined by thin-layer chromatographic scanning(TLCs). RESULTS: The average recovery rate was 97.86%, and the relative standard deviation 1.96%. CONCLUSION: The method is simple, accurate and reliable.