2.Optimization of synthetic pathway and fermentation process of yeast cell factories for production of oleanoic acid.
Dong WANG ; Bei-Bei WANG ; Yi LIU ; Ming-Yu SHI ; Dong-Guang XIAO ; Lu-Qi HUANG ; Zhu-Bo DAI ; Xue-Li ZHANG
China Journal of Chinese Materia Medica 2014;39(14):2640-2645
OBJECTIVETo optimize the synthetic pathway and fermentation process of yeast cell factories for production of oleanoic acid.
METHODUsing the DNA assembler method, one copy of Glycyrrhiza glabra beta-amyrin synthase (GgbAS), Medicago truncatula oleanolic acid synthase (MtOAS) and Arabidopsis thaliana cytochrome P450 reductase 1 (AtCPR1) genes were introduced into Saccharomyces cerevisiae strain BY-OA, resulting in strain BY-20A. YPD medium with different glucose concentration were then used to cultivate strain BY-2OA.
RESULTIncreasing gene copies of GgbAS, MtOAS and AtCPR1 resulted in increased beta-amyrin and oleanolic acid production. The strain BY-2OA produced 136.5 mg x L(-1) beta-amyrin and 92.5 mg x L(-1) oleanolic acid, which were 54% and 30% higher than the parent strain BY-OA. Finally, the titer of oleanolic acid increased to 165.7 mg x L(-1) when cultivated in YPD medium with 40 mg x L(-1) glucose.
CONCLUSIONProduction of oleanoic acid increased significantly in the yeast strain BY-2OA, which can provide the basis for creating an alternative way for production of oleanoic acid in place of extraction from plant sources.
Biomass ; Biotechnology ; methods ; Dose-Response Relationship, Drug ; Fermentation ; Glucose ; pharmacology ; Oleanolic Acid ; biosynthesis ; Saccharomyces cerevisiae ; cytology ; drug effects ; metabolism
3.Role of contrast-enhanced ultrasound in the differentiation of solid focal lesions of pancreas.
Xiao-Yan XIE ; Er-Jiao XU ; Hui-Xiong XU ; Zuo-Feng XU ; Guang-Jian LIU ; Yan-Ling ZHENG ; Jin-Yu LIANG ; Bei HUANG ; Ming-De LÜ
Acta Academiae Medicinae Sinicae 2008;30(1):35-39
OBJECTIVETo investigate the value of contrast-enhanced ultrasound in the differential diagnosis of solid focal lesions of pancreas (s-FLPs).
METHODSWe retrospectively analyzed the clinical data of 56 s-FLPs examined with contrast agent combined with low mechanical indicators contrast-enhanced ultrasound.
RESULTSThe surrounding pancreas parenchyma enhancement time, lesion enhancement time, and peak enhancement time between different groups of s-FLPs had no significant differences (P > 0.05), while the beginning to peak enhancement time showed significant difference (P < 0.05). When using the enhancement speed as a diagnostic indicator to differentiate pancreatic carcinoma from tumor-like pancreatitis, the sensitivity, specificity, and accuracy were 90.5%, 71.4%, and 85.7% for pancreatic carcinoma and 75.0%, 91.7%, and 89.3% for tumor-like pancreatitis. When using the enhancement pattern as a diagnosis indicator to differentiate pancreatic carcinoma from tumor-like pancreatitis, the sensitivity, specificity, and accuracy were 85.7%, 78.6%, and 83.9% for pancreatic carcinoma and 75.0%, 100%, and 94.6% for tumor-like pancreatitis. When different indicators were combined, enhancement pattern and enhancement speed showed the best diagnostic results; however, the Youden index was not improved.
CONCLUSIONSDifferent s-FLPs show different enhancement findings on contrast-enhanced ultrasound. The enhancement pattern and enhancement speed are the most useful diagnostic indicators.
Diagnosis, Differential ; Humans ; Pancreas ; diagnostic imaging ; Pancreatic Neoplasms ; diagnostic imaging ; Pancreatitis ; diagnostic imaging ; Retrospective Studies ; Ultrasonography
4.Cancer incidence and patient survival rates among the residents in the Pudong New Area of Shanghai between 2002 and 2006.
Xiao-Pan LI ; Guang-Wen CAO ; Qiao SUN ; Chen YANG ; Bei YAN ; Mei-Yu ZHANG ; Yi-Fei FU ; Li-Ming YANG
Chinese Journal of Cancer 2013;32(9):512-519
With the growing threat of malignancy to health, it is necessary to analyze cancer incidence and patient survival rates among the residents in Pudong New Area of Shanghai to formulate better cancer prevention strategies. A total of 43,613 cancer patients diagnosed between 2002 and 2006 were recruited from the Pudong New Area Cancer Registry. The incidence, observed survival rate, and relative survival rate of patients grouped by sex, age, geographic area, and TNM stage were calculated using the Kaplan-Meier, life table, and Ederer II methods, respectively. Between 2002 and 2006, cancer incidence in Pudong New Area was 349.99 per 100,000 person-years, and the 10 most frequently diseased sites were the lung, stomach, colon and rectum, liver, breast, esophagus, pancreas, brain and central nervous system, thyroid, and bladder. For patients with cancers of the colon and rectum, breast, thyroid, brain and central nervous system, and bladder, the 5-year relative survival rate was greater than 40%, whereas patients with cancers of the liver and pancreas had a 5-year relative survival rate of less than 10%. The 1-year to 5-year survival rates for patients grouped by sex, age, geographic area, and TNM stage differed significantly (all P < 0.001). Our results indicate that cancer incidence and patient survival in Pudong New Area vary by tumor type, sex, age, geographic area, and TNM stage.
Adult
;
Age Factors
;
Aged
;
Breast Neoplasms
;
epidemiology
;
pathology
;
China
;
epidemiology
;
Colorectal Neoplasms
;
epidemiology
;
pathology
;
Female
;
Humans
;
Incidence
;
Liver Neoplasms
;
epidemiology
;
pathology
;
Lung Neoplasms
;
epidemiology
;
pathology
;
Male
;
Middle Aged
;
Neoplasm Staging
;
Neoplasms
;
epidemiology
;
pathology
;
Pancreatic Neoplasms
;
epidemiology
;
pathology
;
Rural Population
;
Sex Factors
;
Stomach Neoplasms
;
epidemiology
;
pathology
;
Survival Rate
;
Urban Population
5.Reliability of transthoracic echocardiography in estimating the size of Amplatzer septal occluder and guiding percutaneous closure of atrial septal defects.
Gui-shuang LI ; Guang-ming KONG ; Qiu-shang JI ; Ji-fu LI ; Yu-guo CHEN ; Bei-an YOU ; Yun ZHANG
Chinese Medical Journal 2008;121(11):973-976
BACKGROUNDIn China, transthoracic echocardiography (TTE) is popularly used for pre-intervention examination for atrial septal defect (ASD) and for guiding ASD closure. However, the ability to determine ASD size and the safety and efficacy of TTE for guiding ASD closure still has not been widely accepted. This study aimed to evaluate the efficacy and safety of TTE used before, during and after transcatheter ASD closure with Amplatzer septal occluders (ASO).
METHODSSixty-eight subjects (15 men and 53 women; mean age (33.7 +/- 17.3) years) were enrolled. TTE was used to measure the diameters and guide transcatheter closure of ASD. The ASD was examined by long-axis view, basal short-axis view, apical four-chamber view and the subcostal view to observe position, diameter and relation with neighbouring structures. The largest diameter was selected as the reference diameter. Patients were divided into 3 groups according to the ASD reference diameter: 22 subjects with ASD diameter 4 - 14 mm (group A); 21 subjects with ASD diameter 15 - 20 mm (group B); and 25 subjects with ASD diameter 21 - 33 mm (group C).
RESULTSASD was occluded successfully in groups A and B. In group C, occlusion failed in 2 cases; 1 case remained with a 3-mm residual shunt sustained until 6-month follow-up. However, at 6-month follow-up, no case of thromboembolism, ASO dislocation or death occurred in the three groups. The diameter of ASD measured by TTE could accurately predict the ASO size that could successfully occlude the ASD, especially in patients with ASD < 20 mm. The ASD diameter measured by TTE correlated well with ASO size (r = 0.925, P < 0.001; r = 0.976, P < 0.001; r = 0.929, P < 0.001 respectively).
CONCLUSIONSASD diameter measured by TTE can accurately estimate the size of the ASO needed for successful closure of ASD. The larger the ASD, the much larger the ASO needed. TTE is a satisfactory guiding imaging tool for ASD closure.
Adolescent ; Adult ; Aged ; Balloon Occlusion ; instrumentation ; Child ; Child, Preschool ; Echocardiography ; methods ; Female ; Heart Septal Defects, Atrial ; diagnostic imaging ; therapy ; Humans ; Male ; Middle Aged
6.Transfer RNAs inhibit the growth of L929 cells in vitro.
Hong-Mei DING ; Guang YANG ; Hui-Cai CHENG ; Zhao-Hui LIU ; Guo-Jun CAO ; Nong-Le LIU ; Qiang ZHAO ; Ming FAN ; Bei-Fen SHEN ; Ning-Sheng SHAO
Chinese Journal of Applied Physiology 2008;24(3):349-352
AIMTo explore the effects of tRNA on the growth of mammalian cells.
METHODSL929, NIH3T3, MCF-7 and PC12 cells were seeded in 96 well culture plate individually, and incubated at 37 degrees C in 5% CO2 for 4 h, the tRNAs from different species were added to the culture media individually. After certain time of incubation, the viability of the cells was evaluated by the MTT methods. Sub-confluent L929 cells were incubated with 200 microg/ml ytRNA for different times, then the cells were pooled and analyzed with flow cytometry assay.
RESULTStRNA specifically inhibited the growth of L929 cells in a dose-dependent manner. The sizes of tRNA-treated cells showed larger sizes and longer processes than those of untreated cells. Flow cytometric analysis further showed that most of tRNA-treated cells were arrested in S phase of the cell cycle.
CONCLUSIONThe cell growth inhibitory effects of tRNAs were caused mainly by their degraded fragments. The results suggested that tRNA or its degraded fragments might play important roles in regulation of cell proliferation.
Animals ; Cell Cycle Checkpoints ; physiology ; Cell Line ; Cell Proliferation ; Fibroblasts ; cytology ; Flow Cytometry ; Mice ; RNA, Transfer ; physiology
7.Ultrasound-guided percutaneous composite thermal ablation technique in treatment of medium and large hepatocellular carcinoma.
Xiao-yu YIN ; Xiao-yan XIE ; Ming-de LÜ ; Jun-wei CHEN ; Hui-xiong XU ; Zuo-feng XU ; Guang-jian LIU ; Bei HUANG
Chinese Journal of Surgery 2004;42(17):1029-1032
OBJECTIVETo evaluate the clinical values of ultrasound-guided percutaneous composite thermal ablation technique in the treatment of medium and large hepatocellular carcinoma (HCC).
METHODSBy the composite technique of multiple-needle insertion and multi-point energy application, ultrasound-guided percutaneous radiofrequency ablation (RFA) or microwave ablation (MWA) were employed to treat 68 patients of HCC with tumor nodule > or = 3.1 cm in diameter. There was a total number of 73 tumor nodules, with a mean diameter of (4.5 +/- 1.7) cm (3.1-13.0 cm). Sixty-three patients had single tumor nodule, and the remaining 5 had 2 nodules. With a follow-up time of 1.9 - 67.6 months [(16.0 +/- 14.8) months], the local and long-term efficacy was analyzed.
RESULTSHepatic subcapsular hematoma were found in 1 patient. And skin burning in 1 patient 1. Complete ablation rate was 78.1% (57/73), local tumor progression was presented in 24.6% tumor nodules (14/57) and distant recurrence developed in 56.7% patients (38/67). The 1-, 3-, 5-years survival rate was 62.3%, 29.6% and 21.6%, respectively, with a median survival time of 18.6 months, (95% confidence interval: 10.9 months, 26.3 months). There were no remarkable differences in complete ablation rate, local tumor progression, distant recurrence rate and survival rate between RFA and MWA.
CONCLUSIONSBy the composite ablation technique, ultrasound-guided percutaneous thermal ablation was effective in treating medium and large unresectable HCC. And the advantage is minimal invasiveness. RFA and MWA had no marked differences in terms of therapeutic efficacy.
Adult ; Aged ; Carcinoma, Hepatocellular ; diagnostic imaging ; mortality ; surgery ; Catheter Ablation ; methods ; Female ; Follow-Up Studies ; Humans ; Liver Neoplasms ; diagnostic imaging ; mortality ; surgery ; Male ; Middle Aged ; Survival Rate ; Treatment Outcome ; Ultrasonography, Interventional
8.Research on screening and identification of proteins interacting with ataxin-3.
Lu SHEN ; Jian-guang TANG ; Bei-sha TANG ; Hong JIANG ; Guo-hua ZHAO ; Kun XIA ; Yu-hu ZHANG ; Fang CAI ; Li-ming TAN ; Qian PAN
Chinese Journal of Medical Genetics 2005;22(3):242-247
OBJECTIVEThis study sought to isolate and identify the proteins that interact with ataxin-3, to confirm the interacted domain, and to provide new clues for exploring the function of ataxin-3 and the pathogenesis of spinocerebellar ataxia type 3 and Machado-Joseph disease (SCA3/MJD).
METHODSYeast two-hybrid screen (MATCHMAKER GAL4 Two-Hybrid System 3) and regular molecular biologic techniques were undertaken to screen human brain cDNA library with mutant ataxin-3 bait. Two baits from both normal and mutant C-terminus of ataxin-3 were created by subcloned methods to determine which domain of ataxin-3 interacts with the putative associated proteins and to find out optimal candidate proteins that interact with C-terminus of ataxin-3. Confocal microscope was used to observe whether ataxin-3 co-localized with the obtained interacting proteins in mammalian cells.
RESULTSFive novel ataxin-3 interacting proteins were obtained, among which were three known proteins, namely human rhodopsin guanosine diphosphate dissociation inhibitor alpha, small ubiquitin-like modifier 1, and human neuronal amiloride-sensitive cation channel 2; the other two were unknown. Interacting domain analysis revealed that an unknown protein interacted with the C-terminus near the polyglutamine tract of ataxin-3, the other four all interacted with the N-terminus. In the nucleus of SH-SY5Y cell, small ubiquitin-like modifier 1 co-localized with the wild-type ataxin-3 and with the intranuclear aggregates formed by the mutant ataxin-3.
CONCLUSIONAn unknown protein probably interacting with C-terminus of ataxin-3 is firstly discovered, and the initiative findings suggest first that the interaction of small ubiquitin-like modifier 1 with N-terminus of ataxin-3 and the relevant sumoylation probably participate in the post-translation modifying of ataxin-3 and in the pathogenesis of SCA3/MJD.
Acid Sensing Ion Channels ; Ataxin-3 ; Cell Line, Tumor ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Confocal ; Mutation ; Nerve Tissue Proteins ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; Plasmids ; genetics ; Protein Binding ; Recombinant Fusion Proteins ; genetics ; metabolism ; Repressor Proteins ; genetics ; metabolism ; SUMO-1 Protein ; genetics ; metabolism ; Sodium Channels ; genetics ; metabolism ; Transfection ; Two-Hybrid System Techniques
9.Chronic obstructive pulmonary disease assessment test score correlated with dyspnea score in a large sample of Chinese patients.
Qing-tao ZHOU ; Jing-jing MEI ; Bei HE ; Shao-guang HUANG ; Yi SHI ; Fu-qiang WEN ; Ming-wu ZHAO
Chinese Medical Journal 2013;126(1):11-15
BACKGROUNDThe chronic obstructive pulmonary disease assessment test (CAT) is an easy to use health-related quality of life questionnaire, the modified Medical Research Council (mMRC) dyspnea scale is a classic dyspnea scale which is widely used, while the correlation between them is still not clear. This study investigated the use of the Chinese translation of CAT in chronic obstructive pulmonary disease patients and its correlation with the mMRC dyspnea scale.
METHODSThe multicenter cross-sectional study was conducted in 329 hospitals throughout China from March 1 to April 30, 2010. Chronic obstructive pulmonary disease patients completed both the assessment test and the dyspnea scale during a single study visit.
RESULTSSix thousand, four hundred and thirty-seven patients were evaluated; 74.9% were male and the mean age was (64.9 ± 10.0) years. Median test scores in dyspnea grades 0 to 4 were 14, 16, 22, 26 and 32, respectively; these differences were statistically significant. The CAT score was moderately correlated with mMRC dyspnea grade (r = 0.579, P < 0.001). There was no significant difference in mean CAT score between males and females, and patients of high and low socioeconomic status. Primary analysis suggested that CAT scores were higher in older patients (>65 years) than in younger patients (≤ 65 years) and increased with duration of formal education, but these findings were repudiated by further analysis of subgroups according to mMRC dyspnea grade.
CONCLUSIONSThere was no obvious confounding factor influencing use of the CAT in Chinese patients. CAT scores were moderately correlated with the mMRC dyspnea scale.
Adult ; Aged ; Cross-Sectional Studies ; Dyspnea ; psychology ; Female ; Health Status ; Humans ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; psychology ; Quality of Life ; Severity of Illness Index ; Surveys and Questionnaires
10.Incidence and survival condition of primary liver cancer among residents in Pudong district of Shanghai, from 2002 to 2011
Li-Ming YANG ; Xiao-Pan LI ; Chen YANG ; Bei YAN ; Mei-Yu ZHANG ; Yi-Fei FU ; Qiao SUN ; Guang-Wen CAO
Chinese Journal of Epidemiology 2012;33(10):1016-1020
Objective To analyze the incidence and survival rates of primary liver cancer (PLC) among residents in Pudong district of Shanghai.Methods Permanent residents of Pudong district,Shanghai were recruited in the study between 2002 and 2011,among whom 7981 PLC cases were diagnosed and 7233 death cases were certified.Both morbidity and mortality of PLC were calculated and standardized (ASR) by the world population of 1966,in order to analyze the survival condition of PLC patients on gender or address,to explore the prognosis of surgical excision treatment.Results Among the incident cases of PLC between 2002 and 2011,5754 cases were males,with an average age of 60.27; while 2227 cases were females,with an average age of 68.26.The crude incidence rate for both sexes was 30.73 per 100 000 person-year,and was 44.30 per 100 000 person-year for males,higher than that in females,17.16 per 100 000 person-year.The difference showed statistical significance (u=3.46,P<0.01).The ASR for incidence rates were 25.72 per 100 000 person-year in males and 8.22 per 100 000 person-year in females,respectively,with difference statistically significant (u=3.00,P<0.01).There were 7233 PLC death cases in total,including 5151 males and 2082 females,with the crude mortality rate as 27.85 per 100 000 person-year.Rate for males was 39.65 per 100 000 person-year,significantly different from 16.04 per 100 000 person-year in females,statistically (u=3.16,P<0.01).The ASRs for mortality were 22.77 per 100 000 person-year in males and 7.39 per 100 000 person-year in females,with statistically significant difference (u=2.80,P<0.01).Both incidence and mortality rates increased apparently for males aged over 35 and females aged over 45.The 1-to 5-year survival rates of PLC were 19.88%,14.91%,12.19%,10.29% and 8.56%,respectively.There were statistically significant differences among urban,town and rural areas in the over-all survival rates(x2=8.711,P=0.013) ;The differences in 1-to 5-year survival rates between surgical and non-surgical groups showed statistically significant differences (u=10.8,6.93,4.75,3.3 and 2.61,respectively,P<0.01).Conclusion The incidence and mortality rates in males were significantly higher than in females and the over-all survival in rural area was significantly lower than in other areas of Pudong district,suggesting that surgical treatment could improve the prognosis of PLC.