[Investigate optimum conditions and determinate changes of ß-glucosidase activity in Scrophularia root under different drying conditions].

To explore the optimum conditions of ß-glucosidase activity in Scrophularia root by using pNPG method. The extraction conditions and reaction conditions (such as extraction liquid type, reaction system, reaction time, temperature, and substrate concentration) were screened by using monofactorial experiment and homogeneous design. Then the changes of ß-glucosidase activity in Scrophularia root were detected at the drying temperature of 40-100 ℃. The results showed that citric acid phosphate buffer had better extraction effect, and the maximum absorbance produced by enzymatic reaction was present at 50 ℃ environment after reaction for 30 min. Homogeneous design experiment determined that the optimal conditions were as follows: optimal extraction liquid pH 7.0; enzymatic reaction system pH 6.0; substrate concentration 20 mmol•L⁻¹. The change of enzyme activity was affected by drying temperature and water loss rate. In the drying temperature of 60-100 ℃, the enzyme activity was reduced rapidly with the increase in water loss rate, while the activity was seen even with 0% of water at 40 and 50 ℃. This study has laid the theoretical foundation for research of hydrolysis mechanism of iridoid glycosides and optimum drying process.

[A quantitative method for simultaneous assay of seven active ingredients with one marker in Scrophularia ningpoensis root].

To establish a new quantitative method for simultaneous determination of multi-components in Scrophularia root by using high performance liquid chromatography (HPLC) and validate its feasibilities．Meanwhile,using catalpol as one chemical reference substance to establish the relative correct factors and relative retention values of aucubin,harpagide,acteoside,angoroside C,harpagoside and cinnamic acid．Then using the quantitative analysis of multi-components by single-marker (QAMS)model,the six analytes can be quantitatively determined in Scrophularia root．The method was evaluated by comparison of the quantitative results to external standard method．No significant differences were observed between the quantitative results of the two methods．The obtained RCFs were credible．It is feasible and suitable to evaluate the quality of Scrophularia root.

Development of a chemiluminescence enzyme-linked immunosorbent assay for the simultaneous detection of imidaclothiz and thiacloprid in agricultural samples.

A novel and sensitive enhanced chemiluminescence enzyme-linked immunosorbent assay (ECL-ELISA) for the simultaneous analysis of imidaclothiz and thiacloprid is described. The concentrations of coating antibodies and enzyme tracers were optimised by three different ECL-ELISA formats. Under the optimised conditions, when anti-imidaclothiz polyclonal and anti-thiacloprid polyclonal antibodies were immobilised in a single well, single-enzyme tracer ECL-ELISA was used to screen the corresponding pesticide residue, and multi-enzyme tracer ECL-ELISA was used for the analysis of imidaclothiz and thiacloprid residue mixtures. The average recoveries of the two pesticides from spiked tomato, cabbage, and rice samples were in the range of 83.7-117%. Meanwhile, the results showed that the multi-enzyme tracer ECL-ELISA could be applied to the accurate analysis of different proportions of imidaclothiz and thiacloprid (1 : 4 to 4 : 1, m/m) when the gross residues ranged from 20 to 250 µg kg(-1). The results of the multi-enzyme tracer ECL-ELISA in the real tomato samples correlated well with those of high-performance liquid chromatography, with a correlation coefficient of 0.996. Therefore, this new strategy for developing immunoassays is suitable for the simultaneous quantitative detection of imidaclothiz and thiacloprid residues in agricultural samples.

Effects of heat-moisture, autoclaving, and microwave treatments on physicochemical properties of proso millet starch.

Proso millet starch was modified by heat-moisture treatment (HMT), autoclaving treatment (AT), and microwave treatment (MT). The effects of these treatments on the starch physicochemical, structural, and molecular properties were investigated. The amylose and resistant starch contents were increased by AT and MT, but only slightly by HMT. HMT and AT significantly increased the water-holding capacity, to 172.66% and 191.63%, respectively. X-ray diffractometry showed that the relative crystallinity of the HMT sample decreased by 20.88%, and the crystalline peaks disappeared from the AT and MT sample patterns. The thermal treatments decreased the proso millet starch molecular weight to 1.769 × 106, 7.886 × 105, and 3.411 × 104 g/mol, respectively. The thermal enthalpy decreased significantly in HMT. Modification significantly changed the pasting profiles of the native proso millet starch, and the peak viscosity, setback, and breakdown values decreased. These results clarify the mechanism of starch changes caused by thermal treatment.

Chemical Comparison of White Ginseng before and after Extrusion by UHPLC-Q-Orbitrap-MS/MS and Multivariate Statistical Analysis.

Ultrahigh-performance liquid chromatography Quadrupole-Orbitrap tandem mass spectrometry (UHPLC-Q-Orbitrap-MS/MS) was used to compare the composition of ginsenosides in white ginseng (WG) and extruded white ginseng (EWG). A total of 45 saponins, including original neutral ginsenosides, malonyl-ginsenosides, and chemical transformation of ginsenosides, were successfully identified in both WG and EWG. Multivariate statistical analyses including supervised orthogonal partial least squared discrimination analysis (OPLS-DA) and hierarchical clustering analysis (HCA) were used to analyze components of white ginseng before and after extrusion. As a result, three ginsenosides (malonyl (M)-Rb1, M-Rb2, and M-Rc) were found to be increased in WG, while three ginsenosides (Rb2, Rc, and Rg1) were elevated in EWG. In the OPLS-DA S-plot, the different compositions of ginsenoside that were distinguished between WG and EWG were screened out. Experimental results indicate that the UHPLC-Q-Orbitrap-MS/MS is a useful tool to characterize variations of ginsenosides in WG and EWG.

Association of mononucleotide polymorphisms of angiotensinogen gene at promoter region with antihypertensive response to angiotensin receptor blockers in hypertensive Chinese.

INTRODUCTION:: This study aimed to investigate whether mononucleotide polymorphisms of the angiotensinogen gene at promoter were associated with the blood-pressure-lowering response to telmisartan treatment. MATERIALS AND METHODS:: After a two-week single-blind placebo run-in period, 148 patients with mild-to-moderate primary hypertension received monotherapy with 80 mg/day of telmisartan and then were followed up for eight weeks. The -6A/G and -20A/C polymorphisms of the angiotensinogen gene at promoter were determined through polymerase chain reaction and restriction fragment length polymorphsim analysis. The relationship between these polymorphisms and changes in blood pressure was observed and evaluated after eight weeks of treatment. RESULTS:: There were no significant differences between -6A/G, -20A/C polymorphisms of the angiotensinogen gene and blood pressure reductions after treatment, p>0.05. CONCLUSION:: It is suggested that angiotensinogen-6 A/G and angiotensinogen-20 A/C polymorphisms were not associated with the antihypertensive response to telmisartan treatment in Chinese patients with hypertension.

[Prognostic value of microvessel density and angiogenesis-related molecules in epithelial ovarian cancer].

OBJECTIVE: To evaluate the correlations of microvessel density (MVD), vascular endothelial growth factor (VEGF), thrombospodin1 (TSP1) and p53 protein with prognosis in epithelial ovarian cancer. METHODS: Samples from 57 patients with primary epithelial ovarian cancer were examined by immunohistochemical staining using anti-VEGF, anti-TSP1, anti-p53 and anti-CD34 antibodies. The correlation of MVD, expression of VEGF, TSP1 and p53 protein with postoperative recurrence and overall survival were analyzed retrospectively. RESULTS: VEGF, TSP1 and p53 protein was positively detected in 40 (70.2%), 27 (47.4%) and 35 (61.4%) of those patients, respectively. The mean MVD in this series was 30.3 +/- 8.5. High MVD, positive VEGF expression and negative TSP1 expression were positively correlated with postoperative recurrence. Univariate analysis showed that patients with high MVD, positive expression of VEGF and p53 had shorter median overall survival time than those with lower MVD, negative expression of VEGF and p53 (P = 0.0187, P = 0.010 and P = 0.005, respectively), while TSP1 expression was revealed as a protective factor for prognosis. Patients with positive expression of TSP1 had longer median overall survival time than those with negative TSP1 expression (P = 0.042). Multivariate analysis showed that MVD and p53 expression were two independent prognostic factors in epithelial ovarian cancer (P = 0.018 and P = 0.009, respectively). CONCLUSION: VEGF, TSP1 and p53 protein may play an important role in the angiogenesis of epithelial ovarian cancer. High MVD level and p53 protein expression are two independent poor prognostic factors.

[Correlation of preoperative serum vascular endothelial growth factor level with CA125 level in patients with epithelial ovarian cancer and its prognostic value].

OBJECTIVE: To analyze the correlation of preoperative serum vascular endothelial growth factor (VEGF) level with serum CA125 level in patients with epithelial ovarian cancer (EOC), and to evaluate the prognostic value of preoperative serum VEGF in these patients. METHODS: Forty-one patients with EOC were included as study group, while 20 healthy women were selected as control group. Enzyme-linked immunosorbent assay (ELISA) and chemiluminescence assay were used to measure serum VEGF and CA125 level respectively. The correlations of serum VEGF with CA125 level, postoperative recurrence rate and survival time were analyzed retrospectively. RESULTS: Serum VEGF levels in patients with EOC were higher than those in healthy women, with the median of 415 and 165 ng/L, range 110-2120 and 100-735 ng/L respectively (P<0.01). No correlation was found between preoperative serum VEGF and CA125 level (Spearman test, P=0.989). High preoperative serum VEGF was positively correlated with postoperative recurrence. Serum VEGF level in patients with postoperative recurrence was higher than that in patients without recurrence, with the median of 490 and 315 ng/L respectively (P=0.035). Univariate analysis showed that higher serum level was reversely correlated with shorter survival. Median overall survival time in patients with higher serum VEGF level and lower serum VEGF level was 18 months and >35 months respectively (P=0.010). Multivariate Cox model analysis showed that high VEGF level was an independent factor for the prognosis of EOC (P=0.042). CONCLUSION: Preoperative serum VEGF level is not correlated with CA125 concentration in patients with EOC, and it is an independent risk factor for prognosis.

[Study on quality standard of Poria cocos].

OBJECTIVE: To perfect the standard of Poria cocos (Schw.). METHODS: TLC and HPLC fingerprint were used to identify triterpenoids from P. cocos. The content of pachymic acid and dehydropachymic acid were determined by HPLC. RESULTS: Pachymic acid showed a good linear relationship at a range of 1.52 - 7.60 microg/ml, r = 0.9994, the average recovery was 96.90%, RSD = 1.09%. Dehydropachymic acid had a good linear relationship at a range of 0.35 - 1.75 microg/ml, r = 0.9991, the average recovery was 97.86%, RSD = 1.52%. CONCLUSION: The methods are special and accurate, and can be used for the quality control of P. cocos.

Effects of cornstarch on the gel properties of black bean protein isolate.

The effects of common starch (CS) and high amylopectin starch (HAS) from corn on the properties of heat induced black bean protein isolate (BBPI) gels prepared by heating at 95°C for 30 min were investigated by using dynamic oscillatory rheometer, texture analyzer, and scanning electron microscopy (SEM). Compared with BBPI alone, the presence of cornstarch (1-4%, wt/vol) could improve storage modulus (G') and textural properties of BBPI (10%, wt/vol) gels. The mixed system of BBPI and 4% (wt/vol) HAS exhibited the highest G' and formed the gel faster and more easily, which resulted in firmer and more elastic gel than BBPI-CS at all starch concentrations. It was possible that HAS had lower pasting temperature and higher viscosity than CS, which was beneficial to the formation of BBPI gel network and strengthened the stability of network structure. Moreover, it might also be related to the synergistic effect between protein and starch. The CS and HAS existed in the BBPI gel network could bind water, leading to the increase in the water-holding capacity (WHC) of mixed gels, especially 4% (wt/vol) HAS, which was related to homogeneous and compact microstructure with small pores.

[Deletion of p15 and pl6 genes and overexpression of STK15 gene in primary hepatocellular carcinoma].

OBJECTIVE: To investigate the association of p15 and pl6 genes deletion and STKI5 gene overexpression in primary hepatocellular carcinoma (PHC). METHODS: The carcinoma tissue and the adjacent normal tissue were taken from 30 PHC patients during operations who had had neither chemotherapy nor radiotherapy preoperatively. DNA was extracted from the tissues and PCR was used to determine the homozygous deletion of p15 exon2 (pl5E2) and pl6 exon 2 (pl6E2). RNA was extracted, cDNA was synthesized by RT-PCR, and the expression of STKI5 gene was tested by PCR. Beta-actin was used as an internal control. Average density value (ADV) of STK15 gene and that of beta-actin gene were determined in both carcinoma tissue and the adjacent normal tissue. RESULTS: The rate of p15E2 deletion was 13.3% (4/30) and the rate of p16E2 deletion was 16.7% (5/30) in the carcinoma tissue. The p15E2 and pl6E2 co-deletion rate was 6.7% (2/30). In 19 of the 30 cases (63.3%) the expression of STK15 gene in carcinoma tissue was higher than that in the adjacent normal tissue. The ratio of ADV of STK15 gene to ADV of beta-actin gene (1.53+/-0.31) in the carcinoma tissue was significantly higher than that (0.91+/-0.25) in the paired adjacent normal tissue (t = 2.86). CONCLUSION: The homozygous deletion of p15E2 and p16E2 and overexpression of STKI5 gene may play a role in the oncogenesis and malignant progression of PHC.

[Dyuamical studies on metabolic chemistry of lignans from seeds of Arctium lappa].

OBJECTIVE: To study the metabolic chemistry and pharmaco-dynamics characters of ligan from seeds of Arctium lappa. METHOD: HPLC method was used in the study. The analysis was carried out on C18 column. The mobile phase was CH3CN-0.05% H3PO4 (36:64) with flow-rate at 0.6 mL x min(-1) and wave-length of 210 nm. The column temperature was kept at 25 degrees C. RESULT: The results indicated that the ligan was detected in plasma and the main organs 5 min after po. The main metabolic production in plasma was arctigenin. In addition, arctigenin and an unknown product were found in metabolic production in the organs. CONCLUSION: The method was stable,simple and reproducible. It can be used to determine the metabolic product of the ligan. The metabolic chemistry of ligan in plasma was obviously different from that in the main organs.

[Effect of hypoglycosylated E-cadherins on proliferation and invasiveness of tongue squamous cell carcinoma].

PURPOSE: To investigate the effect of N-glycosylated E-cadherins on proliferation and invasiveness of tongue squamous cell carcinoma CAL 27 cells, and to study the effect of hypoglycosylation of E-cadherins on the stability of adherens junctions (AJs) mediated by E-cadherins. METHODS: Human tongue squamous cell carcinoma CAL 27 cells were respectively transfected with the plasmids encoding either hypoglycosylated or wild-type E-cadherins with FLAG as tags. Western blot of FLAG was used to detect the expression of exogenous E-cadherins in the transfected cells after 48 hours. The cell proliferation counting, monolayer cell scratch wound healing and in vitro cell invasiveness assays were performed to evaluate the proliferation and invasiveness of cells. Immunoprecipitation experiments were used to assay the quantity of α-catenins, ß-catenins, γ-catenins and vinculins linked with the cytosolic tails of E-cadherins. All statistical analysis were performed using SPSS 17.0 software package. RESULTS: The expression of exogenous E-cadherins was detected in transfected CAL 27 cells by Western blot. Cell proliferation counting assay revealed that hypoglycosylated E-cadherins significantly inhibited proliferation of CAL 27 cells, compared with wild-type E-cadherins. The monolayer cell scratch wound healing and in vitro cell invasiveness assays indicated that hypoglycosylated E-cadherins also significantly restrained both invasiveness of CAL 27 cells, compared with wild-type E-cadherins. The immunoprecipitation experiments demonstrated that hypoglycosylated E-cadherins exhibited an increased association with α-catenins, ß-catenins, and γ-catenins and vinculins. CONCLUSIONS: Compared with wild-type E-cadherins, hypoglycosylated E-cadherins can significantly suppress proliferation and invasiveness of tongue squamous cell carcinoma CAL 27 cells, and the mechanism was that hypoglycosylated E-cadherins can induce more stable AJs than wild-type E-cadherin. Supported by Scientific Research Foundation for the Returned Oversea Chinese Scholars, State Ministry of Education of China and Shandong Provincial Award Foundation for Excellent Middle- and Young-Aged Scientists (BS2010YY021).