Glycerol is the main byproduct in ethanol production by Saccharomyces cerevisiae. In order to improve ethanol yield and the substrate conversion, a cassette about 4.5 kb for gene homologous recombination, gpd2Δ::PGK1(PT)-POS5-HyBR, was constructed and transformed into the haploid strain S. cerevisiae S1 (MATa) to replace the GPD2 gene by POS5 gene. The NADH kinase gene POS5 was successfully over expressed in the recombinant strain S. cerevisiae S3. Comparing with the parent strain, the recombinant strain S. cerevisiae S3 exhibited an 8% increase in ethanol production and a 33.64% decrease in glycerol production in the conical flask fermentation with an initiatory glucose concentration of 150 g/L. Overexpression of NADH kinase gene seems effective in reducing glycerol production and increasing ethanol yield.
Ethanol ; chemistry ; Fermentation ; Glycerol ; chemistry ; Industrial Microbiology ; Mitochondrial Proteins ; genetics ; metabolism ; Phosphotransferases (Alcohol Group Acceptor) ; genetics ; metabolism ; Saccharomyces cerevisiae ; genetics ; metabolism ; Saccharomyces cerevisiae Proteins ; genetics ; metabolism

OBJECTIVE: To develop a method of quantitative analysis of multi-components by single marker (QAMS) for simultaneous determination of daidzein, daidzin, genistein and genistin in Semen Sojae Praeparatum.

Based on collections and researches of Pesudostellaria heterophylla germplasm resources from different areas of China, by using Shibing SB-4 provenance as materials, the new variety "Shitai No.1" was bred by mass selection, small plot variety comparative test, regional variety comparative test and field trial planting. Compared with "Qian taizishen No.1" and P. heterophylla land races. The disease and lodging resistance, root yield, polysaccharide content and the first grade rate of "Shitai No.1" have obvious advantages. In addition, it is relatively stable of yield in "Shitai No.1" in different places. It is demonstrated that "Shitai No.1" is a fine variety that adapt to the producing areas of P. heterophylla in Guizhou province, it is worthy to be promoted.

OBJECTIVE：To estab lish fingerprint of Duzhong butiansu pill s，analyze its chemical pattern recognition ，and determine the contents of 7 components in Duzhong butiansu pills ，so as to provide reference for the quality control of the preparation. METHODS ：HPLC method was adopted. The determination was performed on Pntulips BP-C 18 Plus column with 0.2％ phosphoric acid water-acetonitrile as mobile phase （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was set at 330 nm，and column temperature was 35 ℃. The sample size was 20 μL. With paeonol as the reference，the HPLC fingerprints of 12 batches of Duzhong butiansu pills （S1-S12） were established with Similarity Evaluation System for TCM Chromatographic Fingerprint （2012 edition）； common peaks were determined and the similarity was evaluated. The chromatographic peaks were identified by comparing with the reference substance. SPSS 21.0 and SIMCA 13.0 software were used for cluster analysis and principal component analysis ，and 22 common peaks were evaluated. The contents of the identified components in 12 batches of samples was determined by the above HPLC method. RESULTS ：A total of 22 common peaks were identified in the HPLC fingerprint of 12 batches of Duzhong butiansu pills ，and the similarity was no loss than 0.960. There were 7 chemical components identified ，which were gallic acid （peak 1），chlorogenic acid （peak 3），liquiritoside（peak 6），hyperoside （peak 7），verbascoside（peak 8），icariin（peak 14）and paeonol （peak 15）. Among the 12 batches of samples ，S1，S3-S5，S7， S9 and S 11 were classified as one category ，S2，S10 and S 124Y091 were clustered into one category ，S6 was one category and S was one category. The 22 common peaks were divided into three principal components. The characteristic value （15.130） and contribution rate （68.775％） of principal component 1 were the largest ，and the score coefficients of peak 3（0.305）and peak 4（0.298）were the highest. Among 12 batches of samples，the cont ents of above 7 components were 18.196 231.951 3，0.000 6-0.049 4，0.234 8-0.415 9，0.039 5-0.079 1，0.053 5-0.249 3，0.000 5-0.000 8，0.646 4-1.146 9 mg/g，respectively. CONCLUSIONS：HPLC fingerprint of Duzhong butiansu pills is established successfully. Twelve batches of samples are clustered into 4 category. Peak 3（chlorogenic acid ）and peak 4（unknown）may be the important factors causing the difference of samples. The content of gallic acid is the highest among the 7 components.

Alcoholic Beverages ; adverse effects ; Animals ; Carcinoma, Hepatocellular ; metabolism ; Gene Expression Regulation, Neoplastic ; drug effects ; Immunohistochemistry ; Liver Neoplasms ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Tumor Suppressor Protein p53 ; metabolism

OBJECTIVETo evaluate the roles of p27(kip1), p16 gene protein and proliferating cell nuclear antigen expression in nasopharyngeal carcinoma (NPC).

METHODSThe EnVision immunohistochemical method was used to detect the expression of p27(kip1), p16 gene protein and PCNA in 66 cases of non-keratinized carcinoma (NKC) and 25 cases of non-tumor nasopharyngeal tissue.

RESULTS(1) The positive expression rates of p27(kip1), p16 gene protein were 65%, 68% in NKC respectively. There were significant differences between NKC and non-tumor group (P < 0.05). (2) The expression of p27(kip1), p16 protein correlated with cranial nerve encroaching and the 5-year survival rates of the patients (P < 0.05), but had no significant correlation to lymph node metastases and clinical staging (P > 0.05). The expression of PCNA was related to clinical staging and to the patient's 5-year survival rates (P < 0.05), but not to lymph node metastases and cranial nerve encroaching (P > 0.05). (3) The positive expression of p27(kip1), p16 gene protein and PCNA were correlated.

CONCLUSIONThe results suggest that immunological labeling of p27(kip1), p16 gene protein and PCNA might be used to determine the prognosis of NKC.

Adult ; Aged ; Cell Cycle Proteins ; analysis ; Cyclin-Dependent Kinase Inhibitor p16 ; analysis ; Cyclin-Dependent Kinase Inhibitor p27 ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; chemistry ; mortality ; pathology ; Prognosis ; Proliferating Cell Nuclear Antigen ; analysis ; Tumor Suppressor Proteins ; analysis

OBJECTIVETo discuss the value of P53mt and BCL-2 proteins in screening skin carcinoma due to arseniasis.

METHODSP53mt and BCL-2 proteins were detected by immunohistochemical staining. chi(2) test was used to analyze the difference of positive rate between two groups. Screening value of the two biomarkers was also evaluated through the analysis of relative indexes.

RESULTSPositive percentages of P53mt and BCL-2 in carcinoma group were 88.89% and 94.44% respectively, both were higher than those of 36.0% and 66.0% in non-carcinoma group (for P53mt, P < 0.01; for BCL-2, P < 0.05). ORs of P53mt and BCL-2 were 14.22 (2.93 - 68.97) and 8.76 (1.07 - 71.51), respectively. Youden's Index and specificity of P53mt were 0.529 and 64.0%, which were much higher than those of BCL-2. Serial tests improved the value of screening with Youden's Index 0.569, but parallel test lowered it to 0.244.

CONCLUSIONSP53mt and BCL-2 were practical biomarkers to screen skin carcinoma due to arseniasis, and the former was better than the latter. The value of screening can be improved by a series of tests.

Arsenic Poisoning ; complications ; Humans ; Immunohistochemistry ; Mass Screening ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Skin ; chemistry ; pathology ; Skin Neoplasms ; diagnosis ; etiology ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

Child ; Child, Preschool ; Female ; Humans ; Infant ; Lymphohistiocytosis, Hemophagocytic ; diagnosis ; drug therapy ; Male ; Practice Guidelines as Topic ; Prognosis

OBJECTIVETo investigate the relationship between the expressions of KAI1, nm23, ETS-1, vascular endothelial growth factor (VEGF) and microvascular density (MVD) and lymph node metastasis and prognosis in nasopharyngeal carcinoma (NPC).

METHODSThe Envision immunohistochemical method was used to detect the expressions of KAI1, nm23, ETS-1 and VEGF in 50 cases of non-keratinizing carcinoma (NKC) with cervical lymph node metastasis, 30 cases of NKC without cervical lymph node metastasis at the primary diagnoses and 30 cases of non-tumor nasopharyngeal tissues (NP). The microvascular density was counted by immunostaining with CD34.

RESULTS(1) The expression rates of KAI1 and nm23 protein in NKC with cervical lymph node metastasis group and without cervical lymph node metastasis group and NP group increased successively , the difference being significant (P < 0.05); The expression rates of ETS-1 and VEGF protein in NKC with cervical lymph node metastasis group and without cervical lymph node metastasis group and NP group increased successively, the difference being significant (P < 0.05). (2) In 80 NKC cases, the MVD was respectively lower in KAI1 and nm23 protein positive groups than those in the negative groups (P < 0.05); the MVD was respectively higher in ETS-1 and VEGF protein positive groups than those in the negative groups (P < 0.05 ). (3) There was significant difference between the MVD, the number of NKC without cervical lymph node metastasis cases in the single expression of KAI1 or nm23 protein and in common expression of KAI1 and nm23 protein (P < 0.05), in the same as between the single expression of ETS-1 or VEGF protein and in common expressions of ETS-1 and VEGF protein (P < 0.05). (4) There was positive correlation between the expressions of KAI1 and nm23 protein (P < 0.01), as well as between the expressions of ETS-1 and VEGF protein (P < 0.01). (5) the 5-year survival rates of the patients correlated with cervical lymph node metastasis and the expressions of KAI1, nm23, ETS-1 and VEGF proteins in NKC (P < 0.05).

CONCLUSIONSThe expressions of KAI1, nm23, ETS-1 and VEGF proteins were highly related to MVD in NPC,cervical lymph node metastasis and prognosis. They might be considered to be reference indicator for evaluating the cervical lymph node metastasis and prognosis of NPC.

Adult ; Aged ; Female ; Humans ; Kangai-1 Protein ; metabolism ; Lymphatic Metastasis ; Male ; Middle Aged ; NM23 Nucleoside Diphosphate Kinases ; metabolism ; Nasopharyngeal Neoplasms ; blood supply ; metabolism ; pathology ; Prognosis ; Proto-Oncogene Protein c-ets-1 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Young Adult

OBJECTIVETo investigate the clinical significance of p16 protein non-expression and p16 gene inactivation by deletions and hypermethylation in nasopharyngeal carcinoma.

METHODSImmunohistochemical study for p16 protein was carried out in 90 cases of non-keratinizing carcinoma (NKC) of nasopharynx. P16 gene deletions and hypermethylation were also analyzed by polymerase chain reaction (PCR) and methylation-specific PCR in 23 randomly selected NKC cases.

RESULTSAmong the 90 NKC cases studied, 42 cases (46.7%) were negative for p16 protein. The non-expression rate of p16 protein also correlated with the 5-year survival rate. The non-expression rate was 60.0% in patients who died within 5 years, in contrast to 20.0% in those alive for over 5 years after diagnosis. The non-expression rate of p16 protein in cases with or without distant metastasis was 81.8% and 41.8% respectively (P < 0.05), while that in cases with or without local invasion into skull base was 41.7% and 48.5% respectively (P > 0.05). As for molecular analysis, deletion of p16 gene exon 2 was found in 10 of the 23 cases (43.4%) studied, while deletion of p16 gene exon 1 was not detected in these samples. Hypermethylation of p16 gene exon 1 was also noted in 2 of the 23 cases (8.7%). The overall mutation rate of these cases was 52.1%.

CONCLUSIONSThere is a high incidence of p16 protein non-expression, deletion of p16 gene exon 2 and hypermethylation of p16 gene exon 1 in NKC. P16 gene inactivation may thus play an important role in the pathogenesis of NKC, especially in terms of its metastatic potential.

CpG Islands ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; DNA Methylation ; DNA, Neoplasm ; genetics ; Exons ; genetics ; Gene Deletion ; Gene Expression Regulation, Neoplastic ; Genes, p16 ; Humans ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; pathology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Skull Base Neoplasms ; pathology ; Survival Rate