1. Expression of human Oct4 and cell penetrating peptide fusion protein
Academic Journal of Second Military Medical University 2010;31(5):489-493
Objective: To express the fusion protein of hOct4 and cell penetrating peptides using prokaryotic expression systems, and to optimize its expression methods and observe the membrane penetrating ability of the fusion proteins. Methods: The pET-based prokaryotic expression system was constructed by genetic engineering, and the fusion plasmid was transferred into E. coli BL21(DE3) and Rosetta2(DE3). The protein was purified by Ni affinity chromatography and identified by Western blotting analysis. The penetrating ability of the Rhodamine-labelled fusion protein was investigated using BJ cells. Results: We successfully constructed pET21a(+)-hOct4-11R-His and pET21a(+)-EGFP-11R-His vectors. Fusion proteins hOct4-11-His and EGFP-11R-His were generated by transfering the plasmids into E. coli. The fusion protein was verified by Western blotting analysis and was detected in BJ cells. Conclusion: We have successfully generated EGFP-11R-His and hOct4-11R-His fusion proteins, and the proteins can effectively enter the BJ cells and locate around the nuclei.
2.Rapid Assessment of Critical Quality Attributes of Chinese Materia Medica:Visualization Technology of Near Infrared Chemical Imaging
Luwei ZHOU ; Zhisheng WU ; Xinyuan SHI ; Yanling PEI ; Yanjiang QIAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(12):2568-2574
Near infrared chemical imaging (NIR-CI) is an emerging technology for rapidly analyzing the critical quality attribute of Chinese materia medica (CMM). It integrates NIR spectroscopy with chemical imaging. In this paper, it provided a systematic introduction to NIR-CI, such as the core part of instrument, the reliability, transformation, analysis and application of high-dimensional data acquisition. In addition, current studies of NIR-CI application in pharmaceutical field were analyzed. Finally, future opportunities and challenges of NIR -CI applications in the quality control of CMM preparation were prospected.
3.Immunity against Schistosoma japonicum Induced by Phage Display Peptides Mimicking Antigenic Epitopes of Trichinella spiralis
Dongming ZHOU ; Xinyuan YI ; Xianfang ZENG ; Min WANG ; Mcreynold LARRY
Chinese Journal of Parasitology and Parasitic Diseases 1997;0(05):-
Objective To obtain the short peptides mimicking antigenic epitopes of Trichinella spiralis ( T\^s\^ ), and explore their cross protective immunity against Schistosoma japonicum ( S\^j. ) in mice. Methods IgG antibodies were purified from sera of mice infected with T\^s\^ . The purified IgG was used to immunoscreen a phage random peptide library of 7 amino\|acid residues displayed as a fusion to protein of filamentous phage. Positive clones were obtained by affinity selection, the reactivity of each clone binding to specific IgG was detected by ELISA. Kunming mice were immunized subcutaneously three times with mixed phage clones. The mice were sacrificed 45 days after challenge. The worms and the liver eggs were counted. Results After three rounds of panning, the relevant phages had been enriched approximately 150 times in production as compared to those from the first round. Of 24 phage clones randomly selected from the third round biopanning, 21 clones were shown to actually bind to the specific IgG. As compared with the control group, the worm and the liver egg reduction rates in vaccination group were 42\^8% and 66\^3% ( P
4.Expression of IL-8 in pulpal tissue injured by maxillofacial firearm trauma
Jun LIU ; Zhizhong CAO ; Zhonghua ZHOU ; Xinyuan WANG ;
Academic Journal of Second Military Medical University 1999;0(12):-
Objective: To provide theoretical basis and experimental evidence for diagnosis and therapy of dental pulp indirect injuries by gunshot wounds. Methods: The levels of IL 8 in different stages were studied using immunohistochemical method to evaluate its functions in inflammation process of dental pulps. Results: IL 8 expression was detected by immunohistochemical assay in postwound conditions. The results showed that positive staining of IL 8 was observed predominantly on infiltrating inflammatory cells, vascular endothelial cells and some fibroblasts, which was consistent with the recruitment and activation of inflammatory cells, and with the severity duration of pulpal tissue injury and response. Conclusion: These results indicate that IL 8 is involved in regulating physiologic responses of dental pulp injuries in maxillofacial trauma by a projectile. [
5.Optimization of Bio-Solid Bidirectional Fermentation Conditions of Trametes Robiniophia Murr . for Rhubarb
Li ZHOU ; Cencong QI ; Pengfei GAO ; Rongrong ZHOU ; Wenyi ZHANG ; Xinyuan SHI
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(11):2500-2505
This study was aimed to optimize bio-solid bidirectional fermentation conditions of Trametes robiniophia Murr. for rhubarb. Sulphuric acid-phenol colorimetry, magnesium-methanol colorimetry, and HPLC were used in the
content determination of polysaccharide, total anthraquinone, and 4 free anthraquinones. The drying rate and con-sumption rate were combined as indicators for the optimization of technical parameters such as medicinal dosage, temperature and amount of water. The results showed that when using 500 mL conical flask, the best fermentation conditions were medicinal dosage of 10 g, the temperature of 34℃, adding water of 120%. It was concluded that bidirectional fermentation of rhubarb increased the content of free anthraquinones. Among them, the content of chrysophanol with anti-oxidation effect increased significantly. The decreasing of combined anthraquinone can relieve the severe laxative effect of rhubarb.
6.Optimization of Near Infrared Variable Selection Method Based on Multivariate Detection Limit
Yanfang PENG ; Xinyuan SHI ; Yang LI ; Luwei ZHOU ; Yanling PEI ; Guodong HUA ; Zhisheng WU ; Yanjiang QIAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(5):960-965
This study was aimed to optimize the near infrared (NIR) variable selection method based on multivariate detection limit (MDL). Using Qing-Kai-Ling (QKL) injection as object, three variable selection methods (interval par-tial least-squares, iPLS; backward interval partial least squares, BiPLS; moving window interval partial least squares, mwPLS) were used to establish the PLS models of baicalin in QKL injection, respectively. The prediction ability of different variable selection method was compared. MDL of all models were calculated in contrast to the MDL value of full spectra PLS model, to select optimal variable selection method. The results showed that different variable selec-tion methods had different prediction ability. Among them, iPLS had the best performance which determination coef-ficient of prediction (Rpre2) and the root mean square errors of prediction (SEP) were 0.996 5 and 602.3 μg·mL-1, re-spectively. All MDLs of different variable selection methods were reduced compared with the full spectra PLS model. The value of iPLS was the lowest comes to be 1.19 μg·mL-1. The results above indicated that the best variable se-lection method for baicalin in QKL injection was iPLS. MDL theory took the error of calibration and validation set and the leverage of external sample into account, which can comprehensively evaluate model detection performance compared to the classic chemical indicator parameters. This method was particularly suitable for the variable selec-tion method optimization of NIR quantitative model of low concentration sample such as Chinese herbal medicine.
7.The suppression of IL-24 on pancreatic cancer mediated by tumor-targeting oncolytic adenovirus in immune-competent mice
Bin XU ; Mingping QIAN ; Tingsong YANG ; Zhenshun SONG ; Xinyuan LIU ; Bo ZHOU
Chinese Journal of Hepatobiliary Surgery 2014;20(6):457-460
Objective To investigate whether IL-24 can inhibit the growth of pancreatic cancer in immune-competent mice,which could be mediated by tumor-targeting oncolytic adenovirus (ZD55-IL-24).Methods Crystal violet staining assay was used to evaluate the anti-proliferative influence of ZD55-IL-24 on Panc02.The tumor growth curve,the tumor formation curve,TUNEL assay,cytotoxic T lymphocyte (CTL) killing assay and serum cytokines detection were used to assess anti-tumor efficacy of ZD55-IL-24 for pancreatic cancer in immune-competent mice.Results ZD55-IL-24 significantly inhibited the growth of pancreatic cancer cells in vitro.Lower tumor formation and slower tumor growth were identified in ZD55-IL-24 group,compared with ZD55-EGFP group or PBS group.ZD55-IL-24 induced pancreatic cancer cells'apoptosis,and mediated anti-cancer immunity by increasing killing responses of CTL to cancer cells and up-regulating serum γ-IFN and IL-6 levels.Conclusion ZD55-IL-24 can induce anti-tumor immunity and suppress pancreatic cancer growth in immune-competent mice.
8.Correlation of sarcopenia and metabolic risk factors
Yu WANG ; Mingxia JIANG ; Qi XU ; Mengmeng WU ; Yinan ZHOU ; Xinyuan DONG ; Jinfeng ZHENG
Journal of Medical Postgraduates 2017;30(2):181-185
Objective Sarcopenia and metabolic syndrome share similar pathophysiological mechanisms. The aim of this study was to investigate the prevalence of sarcopenia among health examination population, and to analyze the relationship between sar-copenia and blood pressure, blood glucose, uric acid and lipids. Methods Physical examination data of 1191 healthy persons in the medical examination center of the hospital from Mar 2011 to Jun 2011 were collected. The weight, skeletal muscle, body fat, body mass index ( BMI) , waist circumference,body fat percentage, waist-hip ratio and visceral fat area were analyzed by human body compositionanalyzer and the prevalence of sarcopenia was observed. At the same time, triglyceride (TG), total cholesterol (TC), high density lipo-protein-cholesterol ( HDL-C ) , low density lipoprotein-cholesterol ( LDL-C) , uric acid and fasting blood glucose were also detected. Results The prevalence rate of sarcopenia of the subjects was 5.21%, and the highest incidence was found in ≥60 years group( 11.11%) . The prevalence rates of overweight and obesity were 33.8% and 10.2%, respectively. The prevalence of sarcopenia is grad-ually higher along with increasing BMI. The prevalence rates of sarcopenia of overweight and obesity subjects were 5.47% and 26.23%, respectively. Compared with the normal control group, the level of weight[(66.34±11.75)kg vs (76.71±12.84)kg ], BMI[(23.37± 3.13) vs (28.05±3.66)], body fat percentage[(25.33±6.06)% vs (36.76±4.47)%], waist circumference[(83.19±9.56)cm vs (95.45±13.74)cm] and visceral fat area[(88.96±29.74)cm2 vs (136.91±25.56)cm2] were higher in the sarcopenia group (P<0.05). Compared with the normal control group, the incidence of systolic blood pressure[(125.59±30.04)mmHg vs (139.39±19.79) mmHg], diastolic blood pressure[(75.82±11.95)mmHg vs (82.34±10.96)mmHg ] TG[(1.56±1.12)mmol/L vs (1.98±1.72)mmol/L] and uric acid[(313.75±83.07)mmol/L vs (335.55±96.07)mmol/L] were higher in the sarcopenia group (P<0.05). Compared with the normal subjects, the detectable rates of abnormal diastolic blood pressure, fasting blood glucose, uric acid, and LDL-C were increased in the sarcopenia, obesity and sarcopenia combined with obesity subjects (P<0.05). The odds ratio of abnormal systolic blood pressure, diastolic blood pressure, uric acid, and LDL-C increased in the sarcopenia, obesity and sarcopenia combined with obe-sity subjects using logistic regression analyses after correction of gender and age. Conclusion The sarcopenia may have some con-nection with metabolic risk factors. Early detection of sarcopenia can help to distinguish people predisposed to metabolic syndrome, and it has important significance for prevention of chronic disease.
9.Study on the Applicability of NIR Combined with LS-SVM Used for the Quality Monitoring of Medicated Leaven Fermentation
Cencong QI ; Zhaozhou LIN ; Rongrong ZHOU ; Wenyi ZHANG ; Pengfei GAO ; Xinyuan SHI
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;17(3):643-647
This article investigated the applicability of the near infrared spectroscopy (NIR) technology, combined with the least squares support vector machines (LS SVM) used for the quality monitoring of medicated leaven fermentation. First, near infrared spectra of 67 medicated leaven samples were obtained by near infrared spectroscopy system in the wavelength range of 400 2 500 nm, and then the protease and amylase activity were measured by Folin phenol method and DNS method. Thereafter, the LS SVM was employed to calibrate models. The Rc and Rp of protease in near infrared model were 0.975 and 0.938, respectively; The RMSEC and RMSEP were 5.297 and 9.795, respectively. The Rc and Rp of amylase in near infrared model were 0.987 and 0.973, respectively; The RMSEC and RMSEP were 7.215 and 6.864, respectively. This model has good prediction ability and is suitable for quality monitoring in medicated leaven fermentation process. The research achievement could lay a certain foundation for the near infrared spectral analysis technology applied in the field of traditional fermentation processing.
10.Primary clinical study of hippocampal-sparing prophylactic cranial irradiation in limited-stage small-cell lung cancer
Xin DONG ; Zongmei ZHOU ; Junjie MIAO ; Xinyuan CHEN ; Zhihui HU ; Peng HUANG ; Yin ZHANG
Chinese Journal of Radiation Oncology 2015;24(2):131-136
Objective To preliminarily observe the clinical efficacy of hippocampal-sparing prophylactic cranial irradiation (HS-PCI) using helical tomotherapy (HT) in patients with limited-stage small-cell lung cancer (LS-SCLC) after chemoradiotherapy,and compare HT with intensity-modulated radiotherapy (IMRT) and volumetric modulated arc therapy (VMAT) in dose distribution.Methods From April to June,2014,six patients with LS-SCLC who had achieved a complete remission after chemoradiotherapy were assigned to HS-PCI using HT within a month after brain metastasis was ruled out using brain magnetic resonance imaging (MRI).After fusing CT images and MRI images,the hippocampus was contoured in the fusion images and hippocampal avoidance regions were created using a volumetric expansion of 3 mm around the hippocampus.A dose of 25 Gy in 10 fractions to 95% of planning target volume (PTV) was prescribed in HT,IMRT,and VMAT.The clinical efficacy,adverse reactions,neurocognitive function,and brain metastasis were evaluated for HT.The dose distribution in PTV and hippocampus were compared between HT,IMRT,and VMAT.Results There were one patient with abdominal wall and abdominal lymph node metastases,one patient with local recurrence,and no patient with brain metastasis during the observation period.The numbers of patients with grade 1 and grade 2 headache,dizziness,and hair loss reactions were 3 and 1,3 and 1,and 4 and 2,respectively.There were no significant differences in the average score of the Mini-Mental State Examination before treatment and at 3 and 6 months after treatment (29.7,29.2,and 29.3 ; P =0.083,0.317,and 0.157).The mean dose to the hippocampus was 16.85 Gy for IMRT and 17.59 Gy for VMAT.For HT,the mean doses to the hippocampus and avoidance regions were reduced to 5.26 Gy and 6.21 Gy,respectively.The prescribed dose for HT was reduced by 79% and 71% compared with IMRT and VMAT,respectively.The average coverage rate of the prescribed dose was 94.48% for HT.Conclusions HT achieves promising dose distribution and target coverage in sparing of the hippocampus.Moreover,HT dose not increase the incidence of adverse reactions.The change in neurocognitive function needs to be further studied with longterm observation and large-scale sampling.