[Effects of tHMGR from three different plant sources on mevalonate (MVA) pathway flux in Saccharomyces cerevisiae].

In this study, we used bioinformatic tools to analyze the 3-hydroxy-3-methylglutaryl-CoA reductase(HMGR) genes from Glycyrrhiza uralensis, Artemisia annua, and Arabidopsis thaliana. The results indicated that GuHMGR and AaHMGR contained two transmembrane regions while AtHMGR had three transmembrane regions. GuHMGR, AaHMGR, and AtHMGR all had the active center for catalysis. Three truncated HMGR genes(tHMGRs) of G. uralensi, A. annua, and A. thaliana were respectively ligated to pYES3 vector to construct the recombinant plasmids pYES3-tGuHMGR,pYES3-tAaHMGR,and pYES3-tAtHMGR. Afterwards, the control plasmid pYES3 and the three plasmids and were respectively introduced into Saccharomyces cerevisiae Cen.pk2-1 D, which yielded strains Y0, Y1, Y2, and Y3, respectively. The content of squalene, lanosterol, and ergosterol in these strains was measured by GC-MS. The relative expression of tGuHMGR, tAaHMGR, and tAtHMGR in strains Y1, Y2, and Y3 was determined by quantitative real-time PCR. The results showed that the strain overexpressing tAaHMGR had the highest yield of squalene and the highest total yield of squalene, ergosterol, and lanosterol. The quantitative real-time PCR showed higher relative expression of tAaHMGR than tGuHMGR, consistent with the strain fermentation result. We selected a superior tHMGR by comparing the effects of different tHMGRs on the mevalonate(MVA) pathway flux in S. cerevisiae. The findings can provide a reference for the construction of S. cerevisiae strains with high yields of squalene and terpenoid precursors.

Manufacture and antibacterial characteristics of Eucommia ulmoides leaves vinegar.

In this work, the fermentation conditions and the antibacterial characteristics of Eucommia ulmoides leaves vinegar (EV) were studied. By single factor orthogonal test, it was found that under optimal fermentation conditions (bran addition 10%, sugar addition 8%, leaven addition 0.3% and acetic acid bacteria solution 12%), the acetic acid content and CA content of EV were 45.5 ± 2.8 mg/mL and 0.98 ± 0.08 mg/mL, respectively. Then, by the disc diffusion method, it was concluded that the antibacterial effect of EV was significantly higher than that of Eucommia ulmoides leaves enzymatic hydrolysate and Zhenjiang aromatic vinegar (P<0.05). An investigation into action mode of EV against Bacillus subtilis indicated that, under the combined action of CA and acetic acid, EV exerted its antibacterial effect by damaging bacterial cell wall and cell membrane, increasing the cell permeability which resulted in the structural lesions and release of cell components, thus led to cell death.

Dibasic Ammonium Phosphate Application Enhances Aromatic Compound Concentration in Bog Bilberry Syrup Wine.

A nitrogen deficiency always causes bog bilberry syrup wine to have a poor sensory feature. This study investigated the effect of nitrogen source addition on volatile compounds during bog bilberry syrup wine fermentation. The syrup was supplemented with 60, 90, 120 or 150 mg/L dibasic ammonium phosphate (DAP) before fermentation. Results showed that an increase of DAP amounts accelerated fermentation rate, increased alcohol content, and decreased sugar level. Total phenol and total flavonoid content were also enhanced with the increase of DAP amounts. A total of 91 volatile compounds were detected in the wine and their concentrations were significantly enhanced with the increase of DAP. Ethyl acetate, isoamyl acetate, phenethyl acetate, ethyl butanoate, ethyl hexanoate, ethyl octanoate, ethyl decanoate, isobutanol, isoamyl alcohol, levo-2,3-butanediol, 2-phenylethanol, meso-2,3-butanediol, isobutyric acid, hexanoic acid, and octanoic acid exhibited a significant increase of their odor activity value (OAV) with the increase of DAP amounts. Bog bilberry syrup wine possessed fruity, fatty, and caramel flavors as its major aroma, whereas a balsamic note was the least present. The increase of DAP amounts significantly improved the global aroma attributes, thereby indicating that DAP supplementation could promote wine fermentation performance and enhance the sensory quality of bog bilberry syrup wine.

Polyphenolic Compositions and Chromatic Characteristics of Bog Bilberry Syrup Wines.

Phenolic compounds determine the color quality of fruit wines. In this study, the phenolic compound content and composition, color characteristics and changes during 6 months of bottle aging were studied in wines fermented with bog bilberry syrup under three different pHs. The total anthocyanins and total phenols were around 15.12-16.23 mg/L and 475.82 to 486.50 mg GAE/L in fresh wines and declined 22%-31% and about 11% in bottle aged wines, respectively. In fresh wines, eight anthocyanins, six phenolic aids and 14 flavonols, but no flavon-3-ols were identified; Malvidin-3-O-glucoside, petunidin-3-O-glucoside and delphinium-3-O-glucoside were the predominant pigments; Chlorogentic acid was the most abundant phenolic acid, and quercetin-3-O-galactoside and myricetin-3-O-galactoside accounted for nearly 90% of the total flavonols. During 6 months of bottle storage, the amounts of all the monomeric anthocyanins and phenolic acids were reduced dramatically, while the glycosidyl flavonols remained constant or were less reduced and their corresponding aglycones increased a lot. The effects of aging on blueberry wine color were described as the loss of color intensity with a dramatic change in color hue, from initial red-purple up to final red-brick nuances, while the pH of the fermentation matrix was negatively related to the color stability of aged wine.

[Comparison of histopathologic changes and expression of biomarkers in breast carcinoma before and after neoadjuvant chemotherapy].

OBJECTIVE: To investigate the relationship between the pathologic responses and histologic type, grade, the expression of ER, PR and HER2 and their changes in breast carcinoma before and after neoadjuvant chemotherapy (NAC). METHODS: Two-hundred and nine cases of breast cancer with NAC were analyzed and clinical, pathologic data were evaluated based on the Miller and Payne ( MP) grading system. The expression of ER, PR and HER2 in the cancers before and after NAC were detected by immunohistochemistry (MaxVision method). SPSS 15.0 software was used to conduct statistical analysis. RESULTS: (1) Pathologic responses to the NAC were graded as MP1 (14 cases), MP2 (35 cases), MP3 (106 cases), MP4 (36 cases) and MP5 (18 cases); (2) The expression of ER in core needle biopsy had related negatively to the pathologic response (chi2 = 33.083, P = 0.001). However, the histologic type, grade, ER and PR status, and HER2 expression in surgically-removed specimens had not related to the pathologic response (P>0.05); (3) After NAC, the pathologic type and grade changed in 6. 8% (9/132) and 34.9% (30/86) of the cases, and the rates of changes in the expression of ER, PR and HER2 were 42.4% (75/177), 55.4% (98/177) and 26.6% (46/173) , respectively. Only the expression of HER2 had significant difference between before and after neoadjuvant chemotherapy (P = 0.049). The changes in other data had no relationship with the pathologic response (P>0.05). CONCLUSIONS: Analysis of core needle biopsy can provide important information to predict the pathologic responses to the NAC. The pathologic appearance, grade, ER, PR and HER2 in breast carcinoma may change after NAC. It is necessary to examine the histologic type, grade and the expression of ER, PR and HER2 after NAC once more.

[Differences between clinical response and pathologic response of breast cancer after neoadjuvant chemotherapy].

OBJECTIVE: to investigate the pathologic basis of the difference between clinical response and pathologic response of breast carcinoma after neoadjuvant chemotherapy. METHODS: two hundred and nine cases of breast cancer with neoadjuvant therapy were analyzed and clinical data were collected from June, 2005 to December, 2007. All patients had core needle biopsy taken before neoadjuvant chemotherapy and were operated within 4 weeks after neoadjuvant chemotherapy. Clinical examination, X-ray of breast and/or B ultrasonography of primary breast focus were taken before and after neoadjuvant chemotherapy. Clinical responses of breast primary focus were evaluated according to RECIST (response evaluation criteria in solid tumors) version 1.1.Pathologic responses of breast primary focus were evaluated according to Miller and Payne (MP) grading system. SPSS 15.0 software was used to statistical analysis. RESULTS: (1) Clinical responses basing on clinical examination showed complete response, partial response, stable disease and progressive response, in 33, 124, 41 and 11 cases respectively. (2) Eighty-seven cases had X-ray of breast taken before and after neoadjuvant chemotherapy. Clinical response basing on X-ray, showed complete response, partial response and stable disease in 8, 42 and 37 cases respectively. (3) Pathologic responses of breast primary focus were as MP1 (14 cases), MP2 (35 cases), MP3 (106 cases), MP4 (36 cases) and MP5 (18 cases). (4) The clinical response basing on clinical examination were related to the pathologic response (χ(2) = 33.668, P = 0.001); and the clinical response basing on X-ray of breast were also related to the pathologic response (χ(2) = 22.404, P = 0.004). (5) The pathologic basis of the difference between the pathologic response and the clinical response basing on X-ray of breast were: embolism of carcinoma, mucinous carcinoma, intraductal carcinoma with ossifying-type calcification, nodular fibrosis and others. CONCLUSIONS: the clinical response may be related to the pathologic response. The difference between the two may be caused by pathologic changes. Some benign and malignant pathologic changes may contribute to the under-estimation of clinical response over pathologic response; whereas embolism of carcinoma may contribute to the over-estimation of clinical response over pathologic response.

[A comparison of localization by ductoscopy-guided wire and conventional methods in terminal duct excision for women with pathological nipple discharge].

OBJECTIVE: To evaluate and compare localization by ductoscopy-guided wire with localization by conventional methods in the terminal duct excision for women with pathological nipple discharge. METHODS: Breast terminal duct excision were performed in 174 consecutive patients with intraductal lesions diagnosed by mammary ductoscopy. Sixty-eight of those underwent ductoscopy-guided wire localization for more accurate ductal excision. The patients received mammary ductoscopy and a hooked wire was anchored at the intraductal lesions under endoscopic surveillance just before the operation. Then a biopsy resection of wire-guided terminal duct and frozen section were done. Tbe other 106 patients received terminal duct excision under localization with conventional methods without ductoscopy either by puncturing a needle or injection of blue dye through the duct with pathological discharge. RESULTS: Of the 68 patients with ductoscopy-guided duct excision, 64 had intraductal papillomas and 4 duct carcinoma in situ proved by pathology. All the lesions in these 68 patients were completely resected during biopsy without extra extended resection, and the concordance rate of the pathological result with ductoscopic diagnosis was 100.0%. None of them developed a postoperative breast distortion. In the conventional method localization group, there were 96 intraductal papilloma, 6 duct carcinoma in situ and 4 adenosis. Only 77.4% of the lesions were excised in the primary biopsy, and 22.6% needed extended resection. The concordance rate of the pathological diagnosis with ductoscopic diagnosis was 96.2%. Twenty-six patients had a deformed breast postoperatively. CONCLUSION: Ductoscopy-guided wire localization is superior to the conventional localization method in the surgical terminal duct excision for women with spontaneous nipple discharge. It is not only helpful for more accurate localization and resection as well as pathologic sampling, but also is minimally invasive. Further studies are still required and this method may deserve to be popularized.

[The quantified analysis of fresh mutton tenderness using PLS methods and Fourier transform near-infrared spectroscopy].

Ninety eight representative fresh mutton samples from Neimeng, Ningxia, Gansu, Xinjiang province were selected for this study, the nondestructive measurement of the fresh mutton tenderness by Fourier transform near infrared (FT-NIR) spectroscopy was discussed. Partial least squares(PLS) algorithm was used to build the model between the shear force value of the fresh mutton tenderness measured by the texture machine and the FT-NIR spectra. The influence of different processing method of spectra, factors and wave regions on the determination coefficients (r2), root mean square error of cross validation (RMSECV) and root mean square error of prediction (RMSEP) was studied. The result showed that the shear force value of ninety eight representative fresh mutton samples was 1.673-6.631 kg, and the shear force value above 75% samples was 2-5 kg, almost covering the fresh mutton tenderness of our country's sheep, the r2 of the calibration could reach 86.2% and the RMSECV was up to 0. 445 in the wave number range 11 995-5 446 cm(-1) and 4 601-4 246 cm(-1) with vector normalization when the PLS factors was ten. The correlation coefficient(R), RMSEP and average bias between value measured by the texture machine and predicted value of model based on validation samples were 0.87, 0.524 and 0.385 respectively. The result indicates that FT-NIR spectroscopy is capable of predicting tenderness value of fresh mutton.