Construction of Eukaryotic Vector Expressing shRNA of Lycopene Cyclase Gene carRA

( Ye Li ),( Lihua Jin ),( Qing Wei ),( Xiaojie Wang ),( Bo Li ),( Yuning Zhang ),( Liqing Zhang ) 조선대학교 공학기술연구원 2012 공학기술논문지 Vol.5 No.2

To construct eukaryotic vector expressing shRNA (short hairpin RNA) of carRA. Methods: Designing three different shRNA targeting the coding sequence of the car RA, the mU6-carRA shRNA was constructed by inserting the designed shRNA to the eukaryotic expression vector mU6pro. The constructed eukaryotic vector expressing shRNA of carRA was transformed into DH5 strain. Finally, the recombinant plasmid identified by restriction endonuclease was used for sequence analysis. Results: It was verified by partial nucleotide sequencing and restriction endonuclease digestion that the constructed eukaryotic vector expressing shRNA of carRA was correct. Conclusion: The results of study lay the foundation for further studying on its inhibitive effect on cycling of lycopene.

Measurement and Correlation of Solubility of Solanesol in Single and Binary Solvent Systems

( Ye Li ),( Xueling Du ),( Jingshun Chen ) 조선대학교 공학기술연구원 2010 공학기술논문지 Vol.3 No.3

The aim of this study was to investigate the solubilities of solanesol in three single solvent and three binary solvent systems at different temperatures ranging from 268.15 K to 303.15 K. The single solvent systems included methanol, ethanol, and acetonitrile. The binary solvent systems included methanol-ethanol, methanol-acetic ester, and ethanol-water. Two empirical equations were proposed to correlate with the experimental solubilities and the calculated results were well consistent with experimental data. The results may provide scientific references for the separation process of solanesol.

A New Model for Predicting the Retention of Stationary Phase in Countercurrent Chromatography

( Ye Li ),( Lishui Sun ),( Xueling Du ) 조선대학교 공학기술연구원 2010 공학기술논문지 Vol.3 No.4

The retention of stationary phase (Sf) is a crucial factor in countercurrent chromatography (CCC), which is used for the derivation of the column efficiency, peak resolution, and solute retention. Both operation conditions (flow-rate of mobile phase and rotation speed) and physical properties of two-phase systems (density difference and viscosity) exert important effects on Sf and have been studied to improve the efficiency in various CCC separations. In this study, an artificial neural network (ANN) was used to simultaneously predict the effects of operation conditions and physical properties of two-phase systems on the retention of stationary phase in CCC. The agreement between the predicted and the literature data, which contained the retention of stationary phase for 16 two-phase systems under different operation condition with 194 data points, was excellent with the total average absolute deviation (AAD) of 1.86% and the maximum deviation of 8.79%. By comparing the results predicted by ANN with the results calculated by the mathematical model proposed in literature, it was found that more accurate predictions were achieved by means of the ANN.

Clonging and Expression of Heparinase III Gene from Flavobacterium heparinum

( Ye Li ),( Donghai Su ),( Lihua Jin ),( Jingjun Wu ),( Xinhui Xing ) 조선대학교 공학기술연구원 2012 공학기술논문지 Vol.5 No.4

To enhance solubility of heparinase III when expressed in recombinant E. coli, the heparinase III gene (HepC) of 1980 bp from Flavobacterium heparinum was amplified by the PCR method, and then it was in inserted in the prokaryotic expression vector pMAL-c2x with the gene of maltose binding protein (MBP) which was identified by colony PCR, restriction enzyme digestion and gene sequencing. The results showed that the fusion strategy by using maltose binding protein (MBP) was effective to enhance solubility when induced at low temperatures, such as 15°C. By shake flask cultivation, the specific enzyme activity of MBP-hepC fusion protein could reach about 46.41 IU/mg which was higher than previous reported results. By one-­step affinity purification with amylose resin, the MBP-hepC fusion protein was easily purified to more than 95% purifty and the enzyme characteristics were studied. The optimum reaction pH value of MEP-hepC was 7.3. The optimum reaction temperature was 42-48°C, which was different from the native heparinase III from Flavobacterium heparinum. This enzymatic reaction to heparine followed Michaelis-Menten equation. The all above realize the process intergration of the enzyme production, separation, purification and applications in LMWH production for reducing the enzyme cost. The study laid the foundation for industrial production of low molecular weight heparin.

Serum microRNA-381: A Potential Marker for Early Diagnosis of Gastric Cancer

Ye Li,Huihui Sun,Jie Guan,Tingting Ji,Xinwei Wang 연세대학교의과대학 2019 Yonsei medical journal Vol.60 No.8

Purpose: The purpose of this study was to explore the potential early diagnostic value of serum microRNA-381(miRNA-381) inpatients with gastric cancer (GC). Materials and Methods: Patients with advanced gastric cancer (AGC) and early gastric cancer (EGC), as well as healthy individuals,were enrolled in this study. Expression of miRNA-381 in serum was detected using real-time quantitative PCR. Electrochemiluminescenceanalysis was used to investigate the expression of classic tumor markers, including carbohydrate antigen (CA) 199,CA724, and carcinoembryonic antigen. Finally, receiver operating characteristic curve and Kaplan-Meier analysis were used todetermine the value of miRNA-381 in clinical diagnosis of GC. Results: miRNA-381 was differentially expressed among the study groups. AUC analysis showed that the sensitivity and specificityof serum miRNA-381 in the diagnosis of GC were superior to those of other tumor markers. Furthermore, low levels of miRNA-381 expression were positively correlated with lymph node metastasis and AGC. Finally, Kaplan-Meier survival analysisshowed that down-regulation of miRNA-381 was associated with lymph node metastasis and the development of GC. Conclusion: miRNA381, which was down-regulated in GC, might be a novel early diagnosis marker for patients with GC.

Treatment of Periodontal Bone Defects with Stem Cells from Inflammatory Dental Pulp Tissues in Miniature Swine

Ye Li,Xi Nan,Tian-Yu Zhong,Tong Li,Ang Li 한국조직공학과 재생의학회 2019 조직공학과 재생의학 Vol.16 No.2

BACKGROUND: Containing a certain proportion of mesenchymal stem cells, inflammatory dental tissue showed great tissue regeneration potential in recent years. However, whether it is applicable to promote tissue regeneration in vivo remains to be elucidated. Therefore, we evaluated the feasibility of stem cells from inflammatory dental pulp tissues (DPSCs-IPs) to reconstruct periodontal defects in miniature pigs. METHODS: The autologous pig DPSCs-IPs were first cultured, appraised and loaded onto b-tricalcium phosphate (b- TCP). The compounds were then engrafted into an artificially-created periodontal defect. Three months later, the extent of periodontal regeneration was evaluated. Clinical examination, radiological examination and immunohistochemical staining were used to assess periodontal regeneration. RESULTS: The data collectively showed that DPSCs-IPs from miniature pigs expressed moderate to high levels of STRO-1 and CD146 as well as low levels of CD34 and CD45. DPSCs-IPs have osteogentic, adipogenic and chondrogenic differentiation abilities. DPSCs-IPs were engrafted onto b-TCP and regenerated bone to repair periodontal defects by 3 months’ post-surgical reconstruction. CONCLUSION: Autologous DPSCs-IPs may be a feasible means of periodontal regeneration in miniature pigs.

Determination of Volatile Components from Grifola frondosa by HS-SPME and GC-MS

( Ye Li ),( Xiao Jie Wang ),( Li Hua Jin ) 조선대학교 공학기술연구원 2012 공학기술논문지 Vol.5 No.3

The head space-solid phase micro extraction-gas chromatograph-mass spectrometry HS-PME-GC-MS was used to analyze the volatile components in Grifola frondosa. The volatile components in Grifola frondosa are extracted by HS-SPME with dry pretreatment then detected by GC-MS. The GC-MS analysis results showed that 56 compositions were identified and accounted for 97.9% of the total peak areas. The major volatile components are esters (51.12%), alkanes (26.97%), aldehyde (7.33%) and terpene (4.90%), respectively. Therefore, HS-SPME-GC-M strategy is a useful and feasible method for the analysis of volatile components in Grifola frondosa.

Plasma CD36 and Incident Diabetes: A Case-Cohort Study in Danish Men and Women

Yeli Wang,Jingwen Zhu,Sarah Aroner,Kim Overvad,Tianxi Cai,Ming Yang,Anne Tjønneland,Aase Handberg,Majken K. Jensen 대한당뇨병학회 2020 Diabetes and Metabolism Journal Vol.44 No.1

Background: Membrane CD36 is a fatty acid transporter implicated in the pathogenesis of metabolic disease. We aimed to evaluate the association between plasma CD36 levels and diabetes risk and to examine if the association was independent of adiposity among Danish population. Methods: We conducted a case-cohort study nested within the Danish Diet, Cancer and Health study among participants free of cardiovascular disease, diabetes and cancer and with blood samples and anthropometric measurements (height, weight, waist circumference, and body fat percentage) at baseline (1993 to 1997). CD36 levels were measured in 647 incident diabetes cases that occurred before December 2011 and a total of 3,515 case-cohort participants (236 cases overlap). Results: Higher plasma CD36 levels were associated with higher diabetes risk after adjusting for age, sex and other lifestyle factors. The hazard ratio (HR) comparing high versus low tertile of plasma CD36 levels was 1.36 (95% confidence interval [CI], 1.00 to 1.86). However, the association lost its significance after further adjustment for different adiposity indices such as body mass index (HR, 1.23; 95% CI, 0.87 to 1.73), waist circumference (HR, 1.21; 95% CI, 0.88 to 1.68) or body fat percentage (HR, 1.20; 95% CI, 0.86 to 1.66). Moreover, raised plasma CD36 levels were moderately associated with diabetes risk among lean participants, but the association was not present among overweight/obese individuals. Conclusion: Higher plasma CD36 levels were associated with higher diabetes risk, but the association was not independent of adiposity. In this Danish population, the association of CD36 with diabetes risk could be either mediated or confounded by adiposity.

Plasma Fetuin-A Levels and Risk of Type 2 Diabetes Mellitus in A Chinese Population: A Nested Case-Control Study

Yeli Wang,Woon-Puay Koh,Majken K. Jensen,Jian-Min Yuan,An Pan 대한당뇨병학회 2019 Diabetes and Metabolism Journal Vol.43 No.4

Background: Fetuin-A is a hepatokine that involved in the pathogenesis of insulin resistance. Previous epidemiological studies have found a positive association between blood fetuin-A and type 2 diabetes mellitus (T2DM) risk among Caucasians and African Americans. We aimed to investigate the prospective relationship between fetuin-A and T2DM in an Asian population for the first time. Methods: A nested case-control study was established within a prospective cohort of Chinese living in Singapore. At blood collection (1999 to 2004), all participants were free of diagnosed T2DM and aged 50 to 79 years. At subsequent follow-up (2006 to 2010), 558 people reported to have T2DM and were classified as incident cases, and 558 controls were randomly chosen from the participants who did not develop T2DM to match with cases on age, sex, dialect group, and date of blood collection. Plasma fetuin-A levels were measured retrospectively in cases and controls using samples collected at baseline. Conditional logistic regression models were used to compute the odds ratio (OR) and 95% confidence interval (CI). Restricted cubic spline analysis was used to examine a potential non-linear association between fetuin-A levels and T2DM risk. Results: Compared with those in the lowest fetuin-A quintile, participants in the highest quintile had a two-fold increased risk of developing T2DM (OR, 2.06; 95% CI, 1.21 to 3.51). A non-linear association was observed (P nonlinearity=0.005), where the association between fetuin-A levels and T2DM risk plateaued at plasma concentrations around 830 μg/mL. Conclusion: There is a positive association between plasma fetuin-A levels and risk of developing T2DM in this Chinese population.

Multiple Biomarkers Improved Prediction for the Risk of Type 2 Diabetes Mellitus in Singapore Chinese Men and Women

Yeli Wang,Woon-Puay Koh,Xueling Sim,Jian-Min Yuan,An Pan 대한당뇨병학회 2020 Diabetes and Metabolism Journal Vol.44 No.2

Background: Multiple biomarkers have performed well in predicting type 2 diabetes mellitus (T2DM) risk in Western populations. However, evidence is scarce among Asian populations. Methods: Plasma triglyceride-to-high density lipoprotein (TG-to-HDL) ratio, alanine transaminase (ALT), high-sensitivity Creactive protein (hs-CRP), ferritin, adiponectin, fetuin-A, and retinol-binding protein 4 were measured in 485 T2DM cases and 485 age-and-sex matched controls nested within the prospective Singapore Chinese Health Study cohort. Participants were free of T2DM at blood collection (1999 to 2004), and T2DM cases were identified at the subsequent follow-up interviews (2006 to 2010). A weighted biomarker score was created based on the strengths of associations between these biomarkers and T2DM risks. The predictive utility of the biomarker score was assessed by the area under receiver operating characteristics curve (AUC). Results: The biomarker score that comprised of four biomarkers (TG-to-HDL ratio, ALT, ferritin, and adiponectin) was positively associated with T2DM risk (P trend <0.001). Compared to the lowest quartile of the score, the odds ratio was 12.0 (95% confidence interval [CI], 5.43 to 26.6) for those in the highest quartile. Adding the biomarker score to a base model that included smoking, history of hypertension, body mass index, and levels of random glucose and insulin improved AUC significantly from 0.81 (95% CI, 0.78 to 0.83) to 0.83 (95% CI, 0.81 to 0.86; P=0.002). When substituting the random glucose levels with glycosylated hemoglobin in the base model, adding the biomarker score improved AUC from 0.85 (95% CI, 0.83 to 0.88) to 0.86 (95% CI, 0.84 to 0.89; P=0.032). Conclusion: A composite score of blood biomarkers improved T2DM risk prediction among Chinese.