1.Screening of proteins interacting with hepatitis C virus core protein from T7-phage display library
Ying HUANG ; Xuefei CAI ; Jun ZHANG ; Ailong HUANG
Journal of Third Military Medical University 2003;0(10):-
Objective To screen and identify proteins that interact with the hepatitis C virus core protein by means of T7-phage display system. Methods The hepatitis C virus core protein was expressed by prokaryotic expression and used as selected molecule to biopan the T7 human liver cDNA library. The selected positive clones were identified by DNA sequence and analyzed with BLAST program in GenBank. Results After BLAST in all positive clones, one protein--Smad interacting protein 1 (SIP1) was found to interact with the hepatitis C virus core protein. Conclusion T7-phage display system is a convenient, rapid and effective method for screening interacting proteins.
5.Safety and feasibility of low?energy X?ray radiotherapy in breast conserving surgery for early?stage breast cancer
Ying XIONG ; Wei ZHOU ; Linping HUANG ; Jun LIU
Chinese Journal of Radiation Oncology 2017;26(3):307-309
Objective To investigate the short?term complications and cosmetic outcomes of radiotherapy in breast conserving surgery for early?stage breast cancer. Methods A retrospective analysis was performed on clinical data from 30 patients with early?stage breast cancer from 2013 to 2015. All patients underwent breast conserving surgery combined with intraoperative low?energy X?ray radiotherapy. The prescribed dose was 20 Gy at the surface of the applicator. Local complications, radiation injury, and cosmetic outcomes were observed after surgery. Results No grade 3?4 adverse reactions were found in patients. In terms of short?term complications, four patients (13%) had seroma, two of whom needed suction treatment;three patients ( 10%) had grade 1?2 skin erythema. Half of the patients had excellent cosmetic outcomes. None of the patients had local recurrence or distant metastasis. Conclusions Intraoperative low?energy X?ray radiotherapy is safe and feasible in breast conserving surgery for breast cancer. It can be considered as an optional approach for tumor bed boost in some patients with early?stage low?risk breast cancer.
6.Analysis of different diagnostic methods for differentiating biliary atresia from hepatitis syndrome
Jun CUI ; Ying HUANG ; Dajia WANG ; Pengjun SU ; Zhibo ZHANG
International Journal of Pediatrics 2014;41(6):652-655
Objective To retrospectively evaluate different methods (laboratory tests,liver / gallbladder ultrasound and magnetic resonance cholangiopancreatography)in differentiating biliary atresia from infant hepatitis syndrome.Methods Seventy infants with cholestatic jaundice,50 cases of biliary atresia and 20 cases of infant hepatitis syndrome were studied prospectively from January 2010 to December 2012.All cases underwent abdominal ultrasound and magnetic resonance cholangiopancreatography.The accuracy,sensitivity,specificity and predictive values of these various methods were compared.Also the laboratory parameters were statistically analyzed and compared between groups.Results Patients with BA had significantly higher GGT values at presentation [(743.5 ± 564) IU/L] compared with infants with IHS [(198.8 ± 197.8) IU/L],showing statistically significant difference (P < 0.05).The sensitivity,specificity,and accuracy of the ultrasound in diagnosis of BA were 84.0% 、100.0% 、88.6% respectively(P < 0.05).The values for magnetic resonance cholangiopancreatography were 82.0% 、80.0% 、81.4% (P < 0.05).Conclusion Currently,the method of ultrasound is more reliable than MRCP for differentiating biliary atresia from infant hepatitis syndrome.
7.Immobilized Lipases Cooperates to Catalyze Transesterification Reaction of Lard
Ying HUANG ; Huan GAO ; Hai ZHEN ; Yun-Jun YAN ;
China Biotechnology 2006;0(01):-
The transesterification reaction conditions of lard with methyl acetate with combined use of immobilized lipases as catalysts were conducted. Initially, according to single factorial experiments, the studies on Lipozyme TL IM and Novozym 435 respectively catalyzed transesterification of lard showed that the optimal parameters of transesterification reaction were: the molar ratio of methyl acetate to oil of 14∶1, 40% enzyme added based on oil weight, temperature 50℃. Combined use of Lipozyme TL IM and Novozym 435 was proposed further to improve the catalytic performance by the response surface method (RSM). Herein, a 5-level-3-factor central composite rotated design was employed to evaluate the effects of lipase loading, the proportion of the two lipases and amount of methyl acetate. The optimum conditions were as followings: 40% lipase loading based on oil weight, 50%/50% the proportion of lipases (Novozym 435/Lipozyme TL IM), and the molar ratio of methyl acetate to oil of 14∶1. And under the optimal conditions, the highest biodiesel yield of 97.6% could be attained, which was higher than the biodiesel yield with each single one of the two lipases. The results suggested that the technics of combined use of certain immobilized lipases catalyzed transesterification reaction of lard for biodiesel production with methyl acetate as the acyl acceptor could raise the FAME yield and save the production cost.
8.Magnetic nanoparticles versus a commercial kit for apolipoprotein E gene polymorphism analysis
Ting WU ; Li YI ; Ying HUANG ; Jun WU
Chinese Journal of Tissue Engineering Research 2013;(38):6804-6810
BACKGROUND:Relative to blood samples, mouth swab samples are more beneficial for apolipoprotein E gene polymorphism analysis among large cohorts. However, agreement has not yet been reached about how to extract genomic DNA form mouth swab samples.
OBJECTIVE:To develop an appropriate method to extract genomic DNA form mouth swab samples, which are suitable for apolipoprotein E gene polymorphism analysis.
METHODS:Fifty mouth swab samples from patients with sporadic Alzheimer’s disease were col ected. Magnetic nanoparticles and PicoDNA trace nucleic acid extraction kit were used to extract genomic DNA form mouth swab samples. And the purity and concentration of the genomic DNA extracted by the two methods were analyzed. Then PCR amplifications and DNA electrophoresis were performed to confirm whether the genomic DNA was able to amplify desired DNA fragments. DNA sequencing was applied to analyze apolipoprotein E gene polymorphisms.
RESULTS AND CONCLUSION:Genomic DNA extracted by the two methods was of high purity. The concentration of genomic DNA extracted by magnetic nanoparticles was higher than by PicoDNA trace nucleic acid extraction kit, and the difference had statistical significance (P<0.05). Al the genomic DNA were able to performed PCR amplifications to obtain desired PCR products, but results of DNA electrophoresis showed that DNA fragments were more clear by nanoparticles method. The results of DNA sequencing were the same by the two methods. The distribution ofε2,ε3,ε4 genotypes of apolipoprotein E gene was 6%, 71%, 23%, respectively. Magnetic nanoparticles were better than PicoDNA trace nucleic acid extraction kit for extracting genomic DNA form mouth swab samples for apolipoprotein E gene polymorphism analysis.
9.Determination of Serum Concentration of Triptolide in Patients with Rheumatoid Arthritis and Its Pharmacokinetics Study
Ying LI ; Yongzhong WANG ; Huan LUO ; Chuanbing HUANG ; Jun ZHU
Chinese Journal of Information on Traditional Chinese Medicine 2014;(1):85-87
Objective To establish an UPLC method for determination of Triptolide in serum and explore its pharmacokinetics in patients with rheumatoid arthritis after oral administration of tripterygium glycosides tablet. Methods Three patients with rheumatoid arthritis were enrolled. Using Estazolam as internal standard, serum was extracted with acetic ether, and determination was performed on column of Waters Acquity C18 (2.1 mm×100 mm, 1.7 μm) with mobile phase consisted of acetonitrile-0.1% glacial acetic acid (30∶70) at the flow rate of 0.2 mL/min. The column temperature was 30 ℃, and the detection wavelength was set at 220 nm. The serum concentration of Triptolide was processed by DAS 2.1.1 computer program. Results Triptolide was well-separated from internal standard, and the retention time were about 4.9 min and 8.9 min, respectively. Linear range of Triptolide was 13.13-840.00 ng/mL. RSD of intra-day and inter-day were lower than 15%and the recoveries were 88.25%-99.33%. Pharmacokinetic parameters were as follows:Cmax was (159.97±42.43) ng/mL, Tmax was (1.33±0.58) h, T1/2βwas (7.51±2.26) h, and AUC0-12 h was (1131.12±89.20) mg?h/L, respectively. Conclusion Pharmacokinetics of Triptolide conformed to two compartment model. Triptolide can be quickly absorbed, and exists differences among individuals.
10.Ease effect of ginsenoside on different-intensity ionizing radiation damage to human hematopoietic stem cells
Ying HUANG ; Xiaoyan LIANG ; Chengjin LI ; Jun HU ; Liqian ZHOU
Chinese Journal of Tissue Engineering Research 2015;(1):124-129
BACKGROUND:Many domestic and foreign scholars and institutions are studying how to relieve radiation damage and to find the most suitable drug, while ginsenosides as the main pharmacological ingredient of ginseng show significant antioxidant effect. OBJECTIVE:To investigate the ease effect of ginsenosides on human hematopoietic stem cels under different intensities of ionizing radiations. METHODS: The CD34+ hematopoietic stem cels were isolated from the healthy cord blood. Then the cels were divided into normal group and ginsenoside-pretreated group, respectively, exposed under 1, 2, 5 Gy of X-ray irradiations for 24 hours. Cel viability was detected in irradiated hematopoietic stem cels by MTT assay. Apoptosis was estimated using the folowing assays: Annexin-V assay, caspase-3 mRNA and protein levels. The generation of reactive oxygen species was evaluated, in the presence or absence of ginsenoside in liquid cultures of CD34+ human hematopoietic stem cels irradiated with 1-, 2- and 5-Gy X-rays, using a flow cytometry assay. The Nrf-2 mRNA and protein levels were also studied by western blot analysis and RT-PCR, respectively. RESULTS AND CONCLUSION: Ionizing radiation at the therapeutic dose could decrease the viability of CD34+ cels and induce the cel apoptosis, and meanwhile, the activity of intracelular reactive oxygen species also showed a progressive increase that was correlated with the dose of ionizing radiation. However, ginsenoside pretreatment could relieve these above-mentioned effects. Ginsenoside inhibited the increase in caspase-3 activity induced by ionizing radiation, and additionaly, enhanced the mRNA and protein expressions of Nrf-2 in CD34+cels. In conclusion, ginsenoside protects CD34+ hematopoietic stem cels from radiation effects, which is probably correlated with anti-apoptosis and anti-oxidant roles of ginsenosides.