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.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
3.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.
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.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.
7.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.
8.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.
9.Improving Effect of Phenylethanoid Glycosides from Tibetan Medicine Phlomis younghusbandii on Rats with Acute High-altitude Cerebral Edema
Fei LUAN ; Maoxing LI ; Rong MA ; Baozhu ZHOU ; Xinyuan CAO ; Yi ZHAO ; Xianmin WANG
China Pharmacy 2015;(22):3075-3078,3079
OBJECTIVE:To investigate the improving effect of phenylethanoid glycosides (PhGCs) from Tibetan medicine Phlomis younghusbandii on rats with acute high-altitude cerebral edema. METHODS:60 Wistar rats were randomly divided into a normoxia control group (isometric sterile water for injection),a hypoxia model group (isometric sterile water for injection),a dexamethasone group(4 mg/kg),and three groups of PhGCs at high(400 mg/kg),middle(200 mg/kg)and low(50 mg/kg)dos-es,with 10 rats in each group. The rats were given drugs,ig,6 d before the establishment of models. On the 4th day of administra-tion,ig,the rats in all groups except the normoxia blank group were placed in a simulated 8 000 m altitude plateau environment for 72 h hypoxic exposure to establish the rat models of high-altitude cerebral edema. Following HE stain,the pathological changes in rats’brain tissues were observed under the light microscope. Dry-wet proportion method was used to determine the water con-tents in rats’brain. The content of MDA and the activities of SOD and GSH in rats’brain tissues were detected. Enzyme-linked im-munosorbent assay was adopted to determine the contents of IL-1β and TNF-α in rats’serum and brain tissues. RESULTS:Com-pared to the rats in the normoxia control group,those in the hypoxia model group showed obvious brain edema,and thickened lacu-nas around cells and vessels and inflammatory cell infiltration, higher water contents and MDA and weaker activities of SOD and GSH in brain,and higher contents of IL-1β and TNF-α in serum and brain tissues. There were statistically significances (P<0.01 or P<0.05). Compared to the rats in the hypoxia model group,those in the groups of PhGCs at high,middleand low dosages demonstrated less inflammatory cell infiltration and lower water contents in brain tissues,in which the groups of PhGCs at high and middle dosages demonstrated lower content of MDA and stronger activities of SOD and GSH in brain tissues, and lower contents of IL-1β and TNF-α in serum and brain tissues. There were statistically significances (P<0.01 or P<0.05). CONCLUSIONS:PhGCs can obviously alleviate the acute cerebral injury in rats which is caused by acute hypoxia and has im-provement effect to some degree on the rats with acute high-altitude cerebral edema.
10.Predictive Value of Microvessel Density and Blood Vessel Invasion in Hepatic Metastasis from Early-stage Rectal Cancer
Yongjian ZHOU ; Huishan LU ; Qin YE ; Guoxian GUAN ; Changming HUANG ; Chuan WANG ; Xinyuan WU ; Xiangfu ZHANG
Journal of China Medical University 2010;(3):205-208,217
Objective To explore the predictive value of microvessel density(MVD)and blood vessel invasion(BVI)in hepatic metastasis from early-stage rectal cancer.Methods MVD and BVI in the tumor tissue from 380 patients with stage I and II rectal cancer was determined by immunohistochemical S-P method with anti-CDIOS antibody and anti-CD34 antibody,respectively.Multinomial logistic regression was performed to analyze the predictive value of MVD and BVI in hepatic metastasis from early-stage rectal cancer.Results CD105 was expressed in newborn blood vessels,not in normal blood veseels.in the rectal cancer tissue.MVD was correlated with histological type and infiltration depth(P<0.05).Besides histological type and infiltration depth,BVI was also correlated with histological grade.Multivariate analysis revealed that histological type,tumor infiltration depth,BVI,adjuvant therapy,and MDV were independent predictors of hepatic metastasis from rectal cancer.The risk of hepatic metastasis in patients with postive expression of either MVD or BVI or both were significant higher than that in patients with low expression of MVD and those without BVI expression[hazard ratio(95%CI),4.210(2.182-11.214)].Conclusion BVI and MVD are independent predictors of hepatic metastasis from stage I and II rectal cancer.Combined detection of MVD and BVI may help to predict the clinical outcome of patients with early-stage rectal cancer.