Effect of RIM21 gene disruption on flocculation of lager yeast / 生物工程学报

Lager yeast is the most popular yeast strain used for beer production in China. The flocculation of yeast plays an important role in cell separation at the end of fermentation. Therefore, appropriately enhancing the flocculation capability of the lager yeast without affecting its fermentation performance would be desirable for beer industry. Our previous study showed that the defect of gene RIM21 might contribute to the enhanced flocculation capability of a lager yeast G03. To further investigate the role of the RIM21 gene in flocculation of strain G03, this study constructed a RIM21-deleted mutant strain G03-RIM21Δ through homologous recombination. Deletion of RIM21 improved the flocculation capability of strain G03 during wort fermentation at 11 °C without changing its fermentation performance significantly. The expression of FLO5, Lg-FLO1 and some other genes involved in cell wall integrity pathway were up-regulated in strain G03-RIM21Δ. In addition, the disruption of RIM21 enhanced resistance of yeast cells to cell wall inhibitors. These results provide a basis for elucidating the flocculation mechanism of lager yeast under low-temperature fermentation conditions.

Regulations of RLM1 gene affect the anti-autolytic ability of lager yeast / 生物工程学报

The autolysis of brewer's yeast seriously affects the quality of beer and the quality of yeast is considered as one of the key factors in beer brewing. Previous studies on brewer's yeast autolysis showed that RLM1 gene, an important transcription factor in cell integrity pathway, is closely related to the autolysis of yeast. In this study, RLM1 was knocked out and overexpressed in a haploid brewer's yeast. RLM1 disruption resulted in poor anti-autolysis performance of yeast, whereas overexpression of RLM1 contributed to the anti-autolytic ability of yeast. In addition, RLM1 gene knockout affected the osmotic stress resistance, cell wall damage resistance, nitrogen starvation resistance and temperature tolerance of yeast strain. The transcriptional level of GAS1 involved in cell wall assembly and DNA damage response was regulated along with the expression of RLM1, whereas other genes in CWI pathway did not show apparent regularity. RLM1 might mainly affect the expression of GAS1 so as to improve the stress resistance of lager yeast in harsh environment. The result from this study help further understand the mechanism of yeast autolysis and lay a foundation for breeding brewer's yeast strain with better anti-autolytic ability.

Progress in brewer's yeast cell wall stress response / 生物工程学报

Yeast cell wall plays an important role in the establishment and maintenance of cell morphology upon the cell wall stress. The cell wall of yeast consists of β-glucans, mannoproteins and chitin. The composition and structure remodel due to cell wall stress. Brewer's yeast cell wall exhibits stress response during long-term acclimation in order to adapt to environmental changes. This paper reviews the composition and structure of yeast cell wall and the molecular mechanisms of cell wall remodeling and signal pathway regulation.

Research progresses in microbial 1,3-1,4-β-glucanase: protein engineering and industrial applications / 生物工程学报

1,3-1,4-β-glucanase (E.C.3.2.1.73) is an important industrial enzyme which cleave β-glucans into oligosaccharides through strictly cutting the β-1,4 glycosidic bonds in 3-O-substituted glucopyranose units. Microbial 1,3-1,4-β-glucanase belongs to retaining glycosyl hydrolases of family 16 with a jellyroll β-sandwich fold structure. The present paper reviews the industrial application and protein engineering of microbial β-glucanases in the last decades and forecasts the research prospects of microbial β-glucanases.

Physiological changes of Saccharomyces cerevisiae during serial re-pitching: a review / 生物工程学报

The morphological structure and physiological indexes of Saccharomyces cerevisiae have changed during serial re-pitching due to the stress conditions in serial handlings and the cells become aging. It is of great significance to study the physiological changes of S. cerevisiae during serial re-pitching to understand the anti-aging effect of S. cerevisiae. In this paper, the changes of the physiological indexes during re-pitching of yeast are summarized, and based on the analysis of the previous works further research directions are proposed.

Study of Risk Factors and Early Diagnosis of Adnexal Torsion / 医学研究杂志

Objective To analysis the risk factors,clinical manifestations of adnexal torsion(AT) during pregnancy,providing evidence for the early diagnosis of AT in pregnancy.Methods Retrospective data were collected from 40 women who underwent surgery for AT during pregnancy between May 2006 and August 2016 at the first affiliated hospital of Wenzhou Medical University.In addition,94 women diagnosed with non-torsion adnexal during pregnancy were chosen to serve as control subjects.The risk factors and clinical characteristics of the two groups wcre compared.Results AT during pregnancy were likely to occurring within the 6th and 14th week of gestation.The rate of the adnexal masses with sizes between 5 and 8cm was significantly higher in torsion group(P ＜ 0.05).The rate of ovarian hyperstimulation syndrome (OHSS) was shown to be obviously elevated in the torsion group (P ＜ 0.05).The white blood cell counts,neutrophil ratio,the rate of lacking of intra-adnexal blood flow on color Doppler imaging were significantly elevated in torsion group (P ＜0.05).Conclusion The risk factors of AT during pregnancy may be occured within the 6th and 14th week of gestation,cyst sizes between 5 and 8cm and combined with OHSS.Adnexal tumors bearing higher risks for torsion and should be strongly considered for an aggressive strategy of management during pregnancy.Moreover,the combined detection of the white blood cell counts and neutrophil ratio as well as doppler imaging can contribute to the early diagnosis of AT during pregnancy.

Advances in anti-staling brewer's yeast / 生物工程学报

Brewer's yeast is crucial in beer fermentation, mainly beer flavor diversity and stability. Beer flavor stability is one of the most influential beer quality aspects, and screening or breeding brewer's yeast with enhanced anti-staling capacity will be an effective solution. In recent decades, with the progress of genetic engineering and detailed description of brewer's yeast genome, great efforts have been made to improve brewer's yeast. This review highlights recent advances in classical and genetic engineering improvement of yeasts to produce more antioxidant compounds or less beer aging substances and precursors. Therein, improvement targets, evaluation methods and development strategies of anti-staling brewer's yeast are also discussed. Furthermore, hotspot and future trend of anti-staling yeast strain development are also proposed.

Human papillomavirus genotype in cervical intraepithelial neoplasia grade Ⅱ/Ⅲ and cervical carcinoma:a comparative study / 实用医学杂志

Objective To explore the distributions of genotypes of HPV infection in CIN and cervical carcinoma. Methods Cervical exfoliated cells were collected from 365 patients with abnormal cervical histology , and subjected to genotyping assay. Results The most prevalent HPV types were 16, 18,52, 58 and 33. The prevalence ratio of HPV 33,52,58 was signi cantly lower in squamous cell carcinoma. Multiple infections decreased from CIN II/III to cervical cancer. Conclusion Besides HPV 16/18, the 52/58/33 subtypes are also important in the development of cervical cancer.

The risk factors analysis of the incidence rate of uroschesis in patients with cervical cancer after extensive hysterectomy / 中国医师进修杂志

Objective To investigate the risk factors of uroschesis in patients with cervical cancer after extensive hysterectomy.Methods The clinical data of 92 patients with cervical cancer who had undergone extensive hysterectomy plus pelvic lymph node dissection from August 2008 to June 2011 were analyzed retrospectively.The incidence rate and related risk factors of uroschesis after operation were analyzed.Results Thirty-one of 92 cases had postoperative uroschesis.The incidence rate of uroschesis was 33.70％.Univariate analysis showed that age,intraoperative blood loss,time of indwelling urinary catheter and urinary tract infection had significant correlation with postoperative uroschesis (P ＜0.05).While operative time,clinical stage,pathological type,preoperative adjuvant therapy,postoperative wound infection or fat liquefaction had no obvious correlation with postoperative uroschesis (P＞ 0.05).Multivariate Logistic regression analysis showed that age,time of indwelling urinary catheter and urinary tract infection were risk factors of postoperative uroschesis.Conclusions Age,time of indwelling urinary catheter and urinary tract infection are independent factors of postoperative uroschesis.The key method to reduce the incidence rate of postoperative uroschesis is to shorten the continuous catheterization time with comprehensive measure,to prevent and control urinary tract infection in early time.

Improvement of thermostability of beta-1,3-1,4-glucanase from Bacillus amyloliquefaciens BS5582 through in vitro evolution / 生物工程学报

In vitro evolution methods are often used to modify protein with improved characteristics. We developed a directed evolution protocol to enhance the thermostability of the beta-1,3-1,4-glucanase. The thermostability of the enzyme was significantly improved after two rounds of directed evolution. Three variants with higher thermostability were obtained. The mutant enzymes were further analyzed by their melting temperature, halftime and kinetic parameters. Comparing to intact enzyme, the T50 of mutant enzymes 2-JF-01, 2-JF-02 and 2-JF-03 were increased by 2.2 degrees C, 5.5 degrees C and 3.5 degrees C, respectively, the halftime (t1/2, 60 degrees C) of mutant enzymes 2-JF-01, 2-JF-02 and 2-JF-03 were shortened by 4,13 and 17 min, respectively, the V(max) of mutant enzymes were decreased by 8.3%, 2.6% and 10.6%, respectively, while K(m) of mutant enzymes were nearly unchanged. Sequence analysis revealed seven single amino acid mutant happened among three mutant enzymes, such as 2-JF-01 (N36S, G213R), 2-JF-02 (C86R, S115I, N150G) and 2-JF-03 (E156V, K105R). Homology-modeling showed that five of seven substituted amino acids were located on the surface of or in hole of protein. 42.8% of substituted amino acids were arginine, which indicated that arginine may play a role in the improvement of the thermostability of the beta-1,3-1,4-glucanase.This study provide some intresting results of the structural basis of the thermostability of beta-1,3-1,4-glucanase,and provide some new point of view in modifying enzyme for future industrial use.

The analysis of upgrade pathologic diagnosis after cervical operation of LEEP(A Report of 101 cases) / 中国医师杂志

Objective To investigate the reasons of upgrade pathologic diagnosis after cervical operation of LEEP and the principal of treatment.Methods 101 cases of clinical data with upgrade pathological diagnosis after LEEP in our hospital during september 2005 to May 2008 were analyzed retrospectively.Results 13 cases were diagnosed as cervical intraepithelial neoplasia(CIN)Ⅰ-Ⅲ while they were considered as chronic cervicitis pre-operation.60 cases of upgrade CIN and 10 cases of in-situ cervical cancer and 10 cases of early infiltration cervical cancer and 8 cases of infiltration cervical cancer were diagnosed while they were considers as CIN Ⅰ-Ⅲ before LEEP.Among the 101 cases.34 cases were re-treated.10 cases diagnosis were the same pathological diagnosis as before,14 cases diagnosis were downgrade and 10 cases were negative.There was no upgrade pathologic diagnosis.The cutting edges were negative.And lymphatic metastasis took place in 2 cases with infiltrating cervical cancer.Conclusion The malignant ratio of patients with upgrade pathology was high.We should pay attention to the patients with upgrade pathological diagnosis after LEEP.The possible treatment should be given to them according to their age,demands of breeding,chnieal stages and types of pathologic stages.

Expression of phosphatidylinositol 3-kinase/protein kinase B signaling pathway in cervical cancer and clinical significance / 中国医师进修杂志

Objective To determine the expression of phosphatidyhnositol 3-kinase (PI3K), phosphorylated protein kinase B (P-AKT) and protein kinase B (AKT) in cervical cancer and explore the correlations with the genesis and development of cervical cancer. Methods Western blot was used to detect the expression of PI3K, P-AKT and AKT proteins in 66 cases of cervical tissues, including 33 cervical cancers, 23 cervical intraepithelial cancer, 10 normal cervical tissues, and analyzed the clinical significances according to clinical informations. Results In cervical cancers, cervical intraepithelial cancer and normal cervical tissues, the expression of PI3K protein were 1.3880±0.0435, 0.5330±0.0939, 0.2427±0.0888 and had significant differences among them, but there was no significant difference in the ratio of P-AKT and AKT, which were 5.8702±0.0543, 5.0755±0.0888, 3.8353±0.0056. The ratios of P-AKT and AKT had association with histological type, the ratios in squamous cancer of the cervix and in adenocancer and denosquamous cancer were 6.7823±0.7745 and 0.7621±0.0709, and the ratios were much higher in squamous cancer of the cervix than adenocancer and denosquamous cancer. Conclusions Overexpression of PI3K protein is involved in the occurrence of human cervical cancer. The ratio of P-AKT and AKT plays a more important role in squamous cancer of the cervix.

Clinical application of human serum kallikrein 6 for the diagnosis and monitor of epithelial ovarian cancer / 中国医师杂志

Objective The aim of this study was to investigate the clinical value of human serum Kallikrein 6 for the diagnosis and monitor of pithelial ovarian cancer. Methods Serum levels of KLK6 were analyzed with ELISA in 30 cases of epithelial ovarian carcinoma, 20 cases of benign ovarian tumor and 30 cases of healthy women. In the meantime, serum CAi25 was determined with chemiluminescence. Furthermore, serum levels of KLK6 and CA125 were also detected in 12 case of epithelial ovarian carcinoma with the same methods one week and the 3rd month postoperation of follow-up. Results Serum levels of KLK6 in epithelial ovarian carcinoma was higher than that in benign ovarian tumor and healthy women (P < 0.05). KLK6 also showed positive correlation with clinical stage, cytological grade, pelvic lymph node metastasis, recurrent or dead disease (P < 0. 05). On the contrary, KLK6 showed no significant correlation with pathological types (P >0. 05). After surgery of follow-up, KLK6 and CA125 were significantly decreased in 12 case of epithelial ovarian carcinoma (P < 0. 05). Furthermore, the total sensitivity and specificity of KLK6 in the diagnosis of epithelial ovarian carcinoma was 73.3% and 85.0% respectively, followed by the sensitivity to be 50. 0% and 88. 9% for the diagnosis of stage Ⅰ-Ⅱand Ⅲ-Ⅳ disease. Conclusion Our resuits showed KLK6 may be one of the reliable indexes for the diagnosis and monitor of ovarian cancer.

Expression and significance of Livin, Survivln in CIN and cervical carcinoma / 中国医师杂志

suitable tumor markers and new targets for gene therapy of cervical carcinoma.

Hormone Therapy on Patients with Amenorrhea Caused by Tripterygium Wilfordii Polyglycoside / 医学研究杂志

Objective To discuss the clinical characteristics and treatment of amenorrhea caused by Tripterygium Wilfordii polyglycoside (TWP).Methods The sexual hormone levels of 25 outpatients of The First Affliated Hospital Of Wenzhou Medical College who suffered from secondary amenorrhea after the use of TWP were examined before and after the hormone therapy.In addition,the clinical presentations were observed.Results (1)The E2 level went down lower than 73.2pmol/L while FSH went up higher than 40U/L in all the 25 patients.Among them,24 patients were confirmed of the diagnosis of premature ovarian failure.⑵HT is an effective way for 20 patients.The total efficiency of the therapy was 80%.⑶The E2 secreted by ovary was higher (63.19?10.81)pmol/l vs. (365.45?161.60) pmol/L and FSH(110.72?21.52)U/L vs. 11.46?17.70)U/Lsecreted by pituitary was lower after therapy than before.⑷After the second period of therapy, the clinical perimenopausal symptoms of all the patients were improved to different degrees.Conclusion TWP may result in the disorders of ovarian functions, but and HT in time may be an effective therapy.

Thoughts on Cultivation of Clinical Teaching in Resident Doctors / 中华医学教育探索杂志

Resident doctors participating in clinical teaching is a disputable topic.We analyze the current situation and difficulties in clinical teaching and propose the concept of RATS(residents as teachers) program in china aiming to strengthen the training of clinical teaching skills in residents and improving culture quality of resident doctors.And we also suggest the establishment of "clinical teaching data warehouses for resident doctors"to keep pace with requirements of our resident training program in new times.

Use of human papilloma virus type 6bL_1 virus-like particles to detect sera antibodies in patients with condylomata acuminata / 中华检验医学杂志

Objective To detect the serum antibody to HPV6 in patients with condylomata acuminata (CA) by use of human papilloma virus(HPV)6bL 1 virus-like particles (VLPS), and to assess the significance of detecting HPV infection. Methods HPV6bL 1 VLPS were produced by a HPV6bL 1 recombinant baculovirus in insect cells,purified by CsCl density gradients centrifugation,and used for ELISA to screen 136 human serum samples in three groups. Results The prevalence of antibody to HPV6bL 1 VLPs was much higher in patients with codylomata acuminata (75%Ab+;mean abs 0.111?0.094) than that in patients with cervical cancer (14.3%Ab+;mean Abs 0.029?0.022)and that in control groups (2.9%Ab+;mean Abs 0.012?0.024). The differences among three groups are significant(P

Relationship of MGB-2 expression and chemotherapy response and prognosis in human ovarian carcinoma / 中国癌症杂志

Background and purpose:In order to look for the new biomarker with higher specificity and sensitivity, we investigated the relationship between expression of MGB-2 and chemotherapy response, and tried to find out whether MGB-2 may sever as a prognostic factor for the patients with ovarian carcinoma. Methods:The expression of MGB-2 was detected by flow-cytometry technology in ovarian carcinoma tissue. We analyzed the correlations among MGB-2 expression and clinical pathological factors, therapeutic effect and prognosis of the patients with ovarian carcinoma.Results:Positive detection of MGB-2 was 78%.(66/885). For the patients whose serum CA125 level did not drop to normal level after two courses of chemotherapy, the MGB-2 positive rate was significantly higher than in those whose serum CA125 level decreased markedly ( 85% vs. 62%, P=0.018). The positive rate was also higher in patients with recurrent disease than in those without recurrence (83% vs. 67%, P=0.097). In univariate survival analysis, the expression of MGB-2 was significantly associated with median survival time (P=0.021).Conclusions:MGB-2 expression is correlated with chemotherapy response and poor prognostic for the patients with ovarian carcinoma.

Study on expression and clinical significance of uPAR and VEGF-C in ovarian cancer / 中国癌症杂志

Background and purpose:urokinase-typeplasminoogen activator receptor(uPAR) plays important roles in terms of fibrolysis in tissue,cell adhering prolifration and signal transduction uPAR can the detected in both cancer and matrix cells.It has been reorted that uPAR is associated with invasion and metastasis of cancer cells.This paper is to study the effects of the gene expression of uPAR and VEGF-C on onco-genesis and progression of ovarian cancer.Methods:The expression of uPAR gene protein and VEGF-C were examined immunohistochemically in 58 cases of ovarian serous cystadenocarcinoma,10 cases of borderline serous cystadenomas and 13 cases of serous cystadenocarcinoma.The relationships between the results and pathological features were studied.Results:① The expressions of uPAR in benign and malignant tumors were different significantly(P