2.Study on hepatocyte cell infected in vitro by hepatitis C virus positive serum
Yan LIU ; Zibai QI ; Heqiu ZHANG ; Hemin LI ; Jinlian GU ; Yang YU ; Zhen YANG ; Kun CHEN
Chinese Journal of Microbiology and Immunology 2008;28(5):440-444
Objective To study the hepatocyte cells infected by hepatitis C virus (HCV) positive serum. Methods Human hepatocyte 7701 was incubated with HCV RNA-positive and HCV antibody(Ab) negative sera BP52. Then, the expression of HCV antigen and the presence of HCV-RNA in cell and supernatant were assayed by RT-PCR, sequence analysis, immunofluorescent staining, Western blot, confocal laser microscopy. The ultrastructural changes of infected cells were observed by electro-microscopy. Results Plus-strand RNA and minus-strand RNA were intermittently detected in cell and/or supernatant on day 7-45 after infection. Sequence analysis demonstrated that the positive DNA nucleic acids were identified with HCV 5′-non-coding region(NCR) sequence. HCV core and NS3 protein were expressed in cytoplasm of infected cells. After 2 or 3 weeks, obvious intracellular ultrastructural changes and virus-like particles were observed. Conclusion human hepatocyte 7701 could support replication of HCV in vitro, which could be a useful tool for setting up cell model of HCV infection and studying the mechanism of HCV infection.
3.Confocal laser endomicroscopy for differential diagnosis of gastric ulcer
Tao LI ; Yanqing LI ; Xiaomeng GU ; Tao YU ; Wenbo LI ; Rui JI ; Zhen HE
Chinese Journal of Digestion 2010;30(1):3-6
Objective To estimate the value of confocal laser endomicroscopy (CLE)for differential diagnosis of gastric ulcer. Methods Patients diagnosed as gastric ulcer by conventional endoscopic mode of CLE were enrolled. The eonfocal endoscopic mode of CLE was used to observe the borderline and/or the surface of the ulcers. Biopsy specimen was taken for histopathological examination and the result of CLE was compared with histopathological finding. Results A total of 12 769 CLE images were obtained from 150 sites of 43 patients. The sensitivity and specificity of CLE for phlegmonosis process were 83.54% and 85.92%, respectively, with positive predictive value of 86.84% and negative predictive value of 82.43%. Whereas the sensitivity and specificity of CLE for intraepithelial neoplasia were 53.57% and 88.52%, respectively, with positive predictive value of 51.72% and negative predictive value of 89.26%. As for diagnosis of gastric cancer, the sensitivity and specificity of CLE were 88.37% and 93.46%, respectively, with positive predictive value of 84.44 % and negative predictive value of 95.24%. The kappa values for agreement of histopathological findings and CLE for diagnosis of phlegmonosis process, intraepithelial neoplasia and gastric cancer were 0.69,0.42 and 0.81, respectively. Conclusions CLE can provide a real-time histopathological diagnosis of the gastrointestinal tract with high sensitivity, specificity, positive and negative predictive values. Targeted biopsy can be done under CLE and the detective rates of precancerous lesions and tumors will be increased.
4.The impact of respiratory motion and active breathing control on the displacement of target area in patients with gastric cancer treated with post-operative radiotherapy
Xiaoli YU ; Zhen ZHANG ; Weilie GU ; Weigang HU ; Ji ZHU ; Gang CAI ; Guichao LI ; Shaoqin HE
Chinese Journal of Radiation Oncology 2010;19(2):131-134
Objective To assess the impact of respiratory motion on the displacement of target area and to analyze the discrimination between free breathing and active breathing control (ABC) in patients with gastric cancer treated with post-operative radiotherapy. Methods From January 2005 to November 2006, 22 patients with post-operatively confirmed gastric cancer were enrolled in this study. All diseases were T_3/ N +, staging Ⅱ - Ⅳ. Patients were CT scanned and treated by radiation with the use of ABC. Image J software was used in image processing, motion measurement and data analysis. Surgical clips were implanted as fiducial marks in the tumor bed and lymphatic drainage area. The motion range of each clip was measured in the resultant-projection image. Motions of the clips in superior-inferior (S-I), right-left (R-L) and anterior-posterior (A-P) directions were determined from fluoroscopy movies obtained in the treatment position. Results The motion ranges in S-I, R-L and A-P directions were 11.1 mam, 1.9 mm and 2.5 mm (F = 85.15, P = 0. 000) under free breathing, with 2.2 mm, 1.1 mm and 1.7 nun under ABC (F = 17.64, P = 0. 000), and the reduction of motion ranges was significant in both S-I and A-P directions (t = 4.36, P = 0. 000;t = 3.73,P = 0.000). When compared with under free-breathing, the motion ranges under ABC were kept unchanged in the same breathing phase of the same treatment fraction, while significant increased in different breathing phase in all three directions (t = - 4.36, P = 0. 000; t = - 3.52, P = 0.000; t =-3.79, P = 0. 000), with a numerical value of 3.7 mm, 1.6 mm and 2.8 mm, respectively (F = 19.46, P = 0. 000) . With ABC between different treatment fractions , the maximum displacements were 2.7 mm, 1.7 mm and 2.5 mm for the centre of the clip cluster (F =4.07,P =0. 019), and were 4.6 mm, 3.1 mm and 4.2 mm for the clips (F =5.17 ,P =0.007). The motion ranges were significant increased in all the three directions (t = - 4.09, P=0.000 ; t =-4.46, P = 0.000 ; t = - 3.45, P =0.000). Conclusions In the irradiation of post-operative gastric cancer, the maximum displacement of organ motions induced by respiration is in S-1 direction and the minimum in R-L direction under free breathing. The use of ABC can reduce the motions significantly in S-I and A-P directions, and the same changes exist in both inter-and intra-fraction treatment.
5.System of Intein-mediated PHB Purified Human Antimicrobial Peptide LL-37
Yin-Xia GU ; Xue-Zhang ZHOU ; Zhen-Wei SONG ; Yu-Jiong WANG ;
China Biotechnology 2006;0(11):-
The new intein-mediated PHB purify protein system is a high expression, automatic cutting, for purification, low-cost protein purification system,it is conducive to large-scale protein purification.Choose human antibacterial peptide LL-37 as the purification objects,which is poison to prokaryotic cell.We construct intein-mediated PHB purified human antimicrobial peptide LL-37 system through genetic engineering technology and use this system to purify LL-37. The results show that this system can highly express LL-37 fusion protein and purifiy the product as same size with expectations.
6.Studies on library information service model:A visualization analysis
Jingli LI ; Yu XIE ; Tianmin ZHEN ; Jingliang GU ; Fang ZHAO ; Zhiyan HAN ; Kuimeng SONG
Chinese Journal of Medical Library and Information Science 2015;(4):27-31
The distribution of authors, institutions that published papers, and key words in CNKI-covered papers on library information service model was analyzed, which showed that there are a number of high production authors in China engaged in studies on library information service model but the number of cooperative studies is few, aca-demic libraries are active in performing related studies but public libraries seldom participate in such studies, the related studies are focused on the application of information technologies and the personal demands of users.
7.A novel full-length gene of human ribosomal protein L14.22 related to human glioma.
Zhen-yu QI ; Guo-zhen HUI ; Yao LI ; Zong-xiang ZHOU ; Shao-hua GU ; Yi XIE
Chinese Medical Journal 2006;119(16):1353-1358
BACKGROUNDThis study was undertaken to obtain differentially expressed genes related to human glioma by cDNA microarray and the characterization of a novel full-length gene.
METHODSTotal RNA was extracted form human glioma and normal brain tissue, and mRNA was used as a probe. The results of hybridization procedure were scanned with the computer system. The gene named 507E08 cone was subsequently analyzed by northern blot, bioinformatic approach, and protein expression.
RESULTSFifteen differentially expressed genes were obtained from human glioma by hybridization and scanning for four times. Northern blot analysis confirmed that the 507E08 clone was low expressed in human brain tissue and over expressed in human glioma tissues. The analysis of BLASTn and BLASTx showed that the 507E08 clone was a novel full-length gene, which codes 203 amino acid of protein and is called human ribosomal protein 14.22 gene. The nucleotide sequence had been submitted to the GenBank with the accession number of AF329277. After expression in E. coli., protein yielded a major band of apparent molecular mass 22 kDa on an SDS-PAGE gel.
CONCLUSIONScDNA microarray technology can be successfully used to identify differentially expressed genes. The novel full-length gene of human ribosomal protein 13.22 may be correlated with the development of human glioma.
Amino Acid Sequence ; Base Sequence ; Blotting, Northern ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Glioma ; genetics ; pathology ; Humans ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; genetics ; metabolism ; Recombinant Proteins ; isolation & purification ; metabolism ; Ribosomal Proteins ; genetics ; metabolism ; Sequence Analysis, DNA
8.The expression of 11 cancer/testis (CT) antigen genes in esophageal carcinoma.
Zhen LIANG ; Zhen-Yuan SUN ; Yan-Hua YUAN ; Yan HAN ; Yu WANG ; Jin GU ; Li-Jian ZHANG
Chinese Journal of Oncology 2005;27(9):534-537
OBJECTIVETo investigate the expression status of 11 different cancer/testis (CT) antigen genes in esophageal carcinoma.
METHODSEsophageal carcinoma tissue and adjacent normal esophageal mucosa taken from 35 esophageal carcinoma patients were assayed for the expression of 11 different CT antigen genes by RT-PCR techniques.
RESULTSOf the 11 CT antigen genes analyzed, none of them was expressed in normal esophageal mucosa. MAGE-3 was found to be the most frequently expressed in esophageal carcinoma tissues (62.9%), followed, in the order of expression frequency, by MAGE4 (31.4%), LAGE-1 (28.6%), MAGE-1 (25.7%), CT10 (20.0%), NY-ESO-1 (20.0%), CT7 (5.7%) and SCP1 (2.9%). No expression of SSX-1, SSX-2 and SSX-4 was found. Among the 35 cases, 28 (80.0%) expressed at least one CT antigen gene, 21 (60.0%) expressed more than 2 CT antigen genes, and 4 of the 21 (19.0%) expressed more than 4 CT antigens, which accounted for 11.4% of total number of patients (4/35). No CT antigen expression was found in the tumor tissue in 7 cases, including 5 cases in stage II and 1 case each in stage I and IV, respectively. Of the 11 CT genes examined, expression of 5 genes (NY-ESO-1, LAGE-1, MAGE-1, MAGE-3 and MAGE-4) was correlated with tumor progression. SCP-1 and CT10 expression was found more frequently in early stage patients. With progression of the disease, the frequency of co-expression of multiple CT antigen genes was significantly increased reaching 28.6% in stage III patients.
CONCLUSIONOf the 11 different CT antigen genes examined by RT-PCR in esophageal carcinoma, 8 genes were detected in various frequencies in 28 of the 35 esophageal cancer patients studied. They are candidate tumor-associated antigens in the preparation of tumor vaccines for immunotherapy in esophageal cancer patients.
Adult ; Aged ; Antigens, Neoplasm ; biosynthesis ; genetics ; Base Sequence ; Esophageal Neoplasms ; genetics ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Neoplasm Proteins ; biosynthesis ; genetics
9.Population pharmacokinetics of high-dose methotrexate administered in 329 children with acute lymphoblastic leukemia.
Chun-Yan ZHANG ; Jian GU ; Yu-Zhen LI ; Wei LU
Journal of Experimental Hematology 2008;16(1):106-110
The aim of this study was to investigate the population pharmacokinetic characteristics of high-dose methotrexate (HDMTX) administered in children with acute lymphoblastic leukemia (ALL) for individualizing methotrexate dose regimen with routine plasma concentration data. The data of 329 children with acute lymphoblastic leukemia administering HDMTX in hospital were collected from 2003 to 2006. Population pharmacokinetics model, parameters, inter-and intra-individual variation were estimated by nonlinear mixed effect modes (NONMEM) software. The effects of the covariates including sex, age, body weight, alkalization and hydration volumes as well as biochemical parameters on the clearance and apparent volume of distribution of methotrexate were explored and the final model was established. The results showed that according to one compartment open pharmacokinetic model with first-order absorption and first-order elimination used in the modeling procedure, the factor that significantly influenced the clearance and apparent volume of distribution of methotrexate was weight (p<0.05). The final model in this study was: CL=7.0 x [1 + 0.0218 x (WT-31.1)] x EXP (eta(CL)) (L/h); V=32.8 x [1 + 0.0288 x (WT-31.1)] x EXP (eta(V)) (L), where 31.1 (kg) was the average body weight of the population. The inter individual variation (CV) of CL and V were 19.4% and 31.0% respectively. It is concluded that the population pharmacokinetic model of HDMTX in children with ALL is established. The obtained final model and population pharmacokinetics parameters of methotrexate may be useful for individualization treatment.
Antimetabolites, Antineoplastic
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adverse effects
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pharmacokinetics
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Antineoplastic Combined Chemotherapy Protocols
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therapeutic use
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Child
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Child, Preschool
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Dose-Response Relationship, Drug
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Female
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Humans
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Male
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Methotrexate
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administration & dosage
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pharmacokinetics
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Models, Biological
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Nonlinear Dynamics
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Precursor Cell Lymphoblastic Leukemia-Lymphoma
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drug therapy
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metabolism
10.The value of confocal laser endomicroscopy for diagnosis of helicobacter pylori infection
Peng WANG ; Rui JI ; Tao YU ; Xiaomeng GU ; Xiuli ZUO ; Chengjun ZHOU ; Changqing LI ; Zhen LI ; Yanqing LI
Chinese Journal of Digestion 2010;30(10):754-758
Objective To investigate the value of fluorescin-aided confocal laser endomicroscopy (CLE) for diagnosis of helicobacter pylori (Hp) infection. Methods From June 2009 to November 2009, patients undergone gastric endoscope examination with upper gastrointestinal symptoms (upper abdominal discomfort, abdominal distension, satiation, acid reflux and eructation) or screened for gastric cancer were enrolled. The gastric mucosa CLE image data of twenty diagnosed Hp positive patients and 10 Hp negative patients was analyzed retrospectively. By comparing with histological image of targeted biopsy tissue, the CLE diagnostic criteria for Hp infection were established. In the prospective study, CLE diagnose result was compared with Hp tested result. The consistency of CLE diagnostic criteria in different observers was also analyzed. The CLE image data with histopathology result were compared accordingly. Results Total 72 patients were enrolled in the prospective study,of 34 Hp positive patients, 31 patients were correctly diagnosed by CLE. The accuracy, sensitivity and specificity of CLE diagnosis were 88.9%, 91.2 and 86.8% respectively. CLE image displaying fluorescin leakage and cell shedding was the highest specificity for Hp infection diagnosis, (97.4 %);fluorescin leakage plus gastric pits distortion and cell edema was the highest sensitivity (88. 2%). The consistency of CLE diagnostic criteria in different observers was high (Kappa value 0. 72, 0.87). The CLE image of Hp infection was highly correlated with inflammation activity (P<0. 001). Conclusion CLE can accurately distinguish normal mucosa from Hp infected mucosa at the cellular level. The diagnostic value for Hp infection was reliable.