1.Identifying the Epitope of Monoclonal Antibody with Phage-displayed Random Peptide Library
Wei SHI ; Hongfu XIE ; Jian LONG ; Xiang CHEN
Chinese Journal of Dermatology 1995;0(01):-
Objective To obtain the short peptides from phage-displayed random peptide library through screening the epitope of monoclonal antibody against tumor necrosis factor(TNF-?). Methods Anti-TNF-? was used to immunoscreen a phage random peptide library of 12 amino-acidresidues displayed as a fusion to protein Ⅲ of filamentous phage M13. The positive clones were obtained by three rounds of biopanning, and the reactivity of each clone binding to anti-TNF-? was examined by double-antibody sandwich ELISA and Dot-ELISA. Mixed positive phage clones were used to detect the serum from SLE patients and healthy persons by Dot-ELISA. Results The eluted phages were enriched nearly 100 fold through three rounds of biopanning, 7 phage clones from the third round biopanning were randomly selected and 5 clones of them could bind to the anti-TNF-?. The binding rate of mixed clones with SLE patients was significantly higher than that of healthy persons. Conclusion The phage display technique can be applied to study the anti-TNF-? antigenic peptides, and these epitopes provide the potential for developing immunodiagnostic reagents of vaccines.
2.Comparison of multi-slice spiral CT portography and endoscopy in the diagnosis of esophageal and gastric varices in liver cirrhosis
Xin HE ; Zhongkui HUANG ; Liling LONG ; Qijun WEI ; Xiaohong JIANG ; Pingping GUO ; Chao XIANG ; Jiang LAN
Chinese Journal of Radiology 2012;(12):1092-1095
Objective To study the correlation of multi-slice CT portography (MSCTP) and digestive endoscopy in the diagnosis and evaluation of esophageal and gastric varices (EGV) caused by cirrhosis.Methods A total of 92 patients with cirrhosis were enrolled in the prospective study.All the patients were examined by endoscopy and 64-slice spiral CT scan in 4 weeks.The types,grading of EGV were observed by endoscopy and MSCTP,and Kappa conformance test was applied with the endoscopic findings as gold standard.The sensitivity,specificity,consistency,and Youden index were evaluated for the diagnosis of sophageal and gastric varices by MSCTP.Results Sixty-five patients were diagnosed to have EGV by endoscopy and 27 were negative.The positive patients included 45 patients of GOV1,19 of GOV2 and 1 patient of IGV1.MSCTP diagnosed 67 cases of EGV and 25 patients of negative results.The positive patients included 46 of GOV1,18 of GOV2 and 3 of IGV1.Two patients of IGV1 varicose veins without positive findings on endoscopy were diagnosed by using MDCTP,which revealed isolated varicose veins under the gastric mucosa.There was high consistency between MSCTP and EGV in the diagnosis of EGV (Kappa =0.732,P < 0.01).The sensitivity of MSCTP was 93.8%,specificity was 77.8%,consistency was 89.1%,and Youden index 71.6%.There was high consistency between MSCTP and EGV in the classification of EGV (Kappa values were 0.743 and 0.763,P < 0.01).Conclusions There is high consistency between MSCTP and digestive endoscopic in the diagnosis and classification of EGV in cirrhosis.MSCTP is superior to endoscopy in the detection of gastric varices.
3.High-intensity pulsed electromagnetic field induces the differentiation of neural stem cells from neonatal rats into neurons in vitro
Ting JIANG ; Tao XU ; Wei XIANG ; Xinlan LONG ; Bingbing LI ; Tao PENG ; Quqin SUN
Chinese Journal of Physical Medicine and Rehabilitation 2014;36(10):740-744
Objective To Investigate the influence of high-intensity pulsed electromagnetic field (HIPEMF) on neural differentiation of neonatal rats neural stem cells in vitro.Methods Neural stem cells (NSCs) were isolated from the subventricular zone of 3-day-old neonatal rats and cultured with serum-free condition medium for 14 days.All the NSCs were then randomly classified into an experimental group which received the stimulation of HIPEMF (0.1 Hz,4 T,8 pulses) and a control group which received no special intervention.Differentiation of the culture was induced by addition of 10% fetal bovine serum on the first day after intervention,the morphological changes of cells were observed under the microscope at different time points.The NSCs adhered to the wall and differentiated for seven days,the immunofluorescence was employed to observed and calculate the ratio of differentiated cells with astrocyte marker GFAP or neuronal markers TUJ1.RT-PCR and western blotting were used to measure the expression levels of the differentiated cells based on gene and protein levels,respectively.Results Immunofluorescence staining showed the number of TUJI positive cells in the experimental group(33.4% ± 5.1%)was significantly more than the control group (26.5% ± 7.0%),while the number of GFAP positive cells was decreased(23.9% ± 5.0%) as compared with the control group(36.2% ± 2.2%).RT-PCR showed that the TUJ1 mRNA expression levels in the experimental group was (1.682 ± 0.086) times of the control group.Western blot showed that the expression of TUJ1 (0.729 ±0.061) in the experimental group was higher than in the control group (0.590 ± 0.157),while the expression of GFAP in the experimental group (0.566 ± 0.056) was less than in the control group(1.034 ± 0.051).Conclusions HIPEMF facilitates differentiation of neural stem cells to neurons,at the cost of reducing astrocytic differentiation.
4.The role of galactomannan detection in the diagnosis of invasive pulmonary aspergillosis in critically ill patients
Yan SHI ; Dawei LIU ; Yun LONG ; Ye LIU ; Xi RUI ; Xiang ZHOU ; Xiaoting WANG ; Wei DU
Chinese Journal of Internal Medicine 2009;48(3):225-230
Objective To evaluate the usefulness of serum galactemannan(GM) for the diagnosis of invasive pulmonary aspergillosis (IPA) in critically ill patients. Methods Study was conducted between February 2007 and July 2008. Included patients on admission ICU who suffer from suspected IPA. GM test and culture were collected 2 weekly. Chnical feature, mycological evidence and optical density index (ODI) were noted. Clinically invasive fungal infection(IFI) were defined proven, probable and possible. The patients were classified into neutropenia, non-neutropenia and treated with immunosuppressive agents, non-neutropenia and non-immunosuppressive agents. To compared of the sensitivity and specificity of GM in different patients. Results 94 patients were included, 4 patients were proven, 29 patients were probable, 34 patients were possible IFI, 27 patients were non-IPA. The positive rate of the GM was 31.9% (30/94). The sensitivity and specificity of GM in proven cases and probable cases are 66.7% and 92.6%. GM assay tended to become positive earlier than the culture 2-10(5.33±2.17)d. We found that differences in patient diagnosis and selection might account for the disparities seen for positive rate for the GM test. There was positive in three of the four patients with proven, the positive rate of GM was 65.5% for probable cases, for possible cases was 17.6%, for non-IPA cases was 7.4% (P=0.001). For patient with neutropenia , treated with immunosuppressive agents and without immunosuppressive agents, the positive rate of GM was 52.9%vs 41.7% vs 34. 6% (P=0.015) ;the sensitivity was 80.0% vs 70. 0% vs 53.8% (P=0.011), the ODI was 1.365 (0.582-6.736) vs 1. 123 (0. 623-6.868) vs 0.554 (0.522-0.823), P=0. 005, respectively. Conclusion These results show that GM test is useful for early diagnosis IPA in critically ill patients. Differences in patient selection and diagnosis might account for the disparities seen for positive rate and sensitivity for the GM test. It has been higher sensitivity and ODI in the patient treated by immunosuppressive agents.
5.Effect of typeⅢsecretion system deficient mutant of E.coli O157∶H7 on attachment of HeLa cells
Yurui WANG ; Xiang LIAO ; Wei ZHOU ; Hongmei DAI ; Ting SONG ; Long LIANG
Military Medical Sciences 2014;(10):795-798
Objective To construct the typeⅢ secretion system (T3SS) deficient mutant of O157∶H7 EDL933, and to detect its ability of attachment after infection with HeLa cells.Methods The recombinant DNA fragments obtained kana-mycin resistant gene were constructed by overlap extension PCR and transferred into EDL933/pKD46 to replace escR gene by homologous recombination.Results The T3SS-deficient mutant (ΔescR) was successfully construted.Compared with the wild-type EDL933, the growth rate and attachment on HeLa cells of ΔescR were significantly decreased.Conclusion The deletion of escR gene, which encodes T3SS structural protein EscR, affects the attachment of EDL933, suggesting a better basis for future studies of T3SS.
6.Anti-mutated citrumnated vimentin antibodies in the diagnosis of rheumatoid arthritis
Hong ZHANG ; Xiang-Pei LI ; Guo-Sheng WANG ; Long QIANG ; Wei WANG ; Lei CHEN ;
Chinese Journal of Rheumatology 2003;0(07):-
Objective To determine the diagnostic value of anti-mutated citrullinated vimentin(anti- MCV)antibodies for rheumatoid arthritis(RA).Methods The anti-MCV were determined in 136 patients with RA,80 non-RA patients and 19 normal peoples.The diagnostic value of anti-MCV was assessed and compared with anti-CCP,AKA and RF.Results The sensitivity and specificity of anti-MCV in the 136 RA patients was 95.6% and 80.8% respectively,there was significanl difference between the test group and the control group(P
7.In vitro study of different Chinese herbs on the proliferation and COMP expression of chondrocyte
Yue-Long CAO ; Wei FENG ; Xiang WANG ; Yu XU ; Hong-Sheng ZHAN ; Yin-Yu SHI ;
Chinese Journal of Rheumatology 2003;0(08):-
Objective To investigate the effect of different Chinese herbs on cell proliferation and cartilage oligomeric matrix protein(COMP)expression in chondrocyte culture.Methods Chondrocytes isolated from rabbit knee cartilage were cultured for 3 generations with the density of 2?10~4/cm~2 and were verified by collagenⅡimmunohistochemical staining.Rabbit sera containing herbs were obtained after animals orally ad- ministrated herbs at the dosage equivalent to human.At 5% and 10% serum density,cells were cultured in the medium that contained liver-softening herbal compound sera.Subgroups setting at 1,3 and 5 hours after herb intervention were observed.Rabbit and bovine sera were control groups.Seven days after intervention,chon- drocytes proliferation was observed using the MTT assay kit.For the study of COMP expression,chondrocytes were isolated from human knee cartilage supematant.Superuatant COMP level was tested by enzyme-linked immunoabsorbent assays(ELISA)after directly adding compound and extract from liver-softening herbs to the culture at the final concentration of 10 mg/ml for 3 days.Results Liver-softening herbal compound group had significant effect on cell proliferation compared to control,of which,3-hour subgroup was more significant than 1-and 5-hour subgroups(P
8.The relationship between lumbar disc degeneration of lumbar spinal stenosis and the dural sac cross-sec-tional area by MRI measurement
Xiang LIU ; Feng WANG ; Xiaodong ZHAO ; Zhirui YAN ; Wei JI ; Jixing WANG ; Yaowu LONG
The Journal of Practical Medicine 2017;33(14):2332-2335
Objective To explore the relationship between lumbar disc degeneration (LDD) of lumbar spinal stenosis(LSS)and the dural sac cross-sectional area(DSCA)by MRI measurement. Methods 91 patients with central degenerative LSS were randomly selected and 91 age-and sex-matched people without LSS were select-ed as a control group. LDD was classified into five grades by MRI detection according to the method proposed by Pfirrmann and DSCA were measured. Results LDD was not associated with age in LSS. The proportion of severe degenerated disc in lower lumbar levels were higher than that of L2/3 in the two groups;DSCA in severe degenerat-ed disc group was significantly smaller than that in light degenerated group only in L2/3 and L3/4 in LSS. There were no statistical differences in every lumbar level in the control group. Conclusions LDD in L4/5 and L5/S1 of LSS is more severe than that of the normal people. DSCA and LDD are positively correlated in L2/3 and L3/4,but not in L4/5 and L5/S1 for LSS.
9.The expression of nicotinamide phosphoribosyl transferase and vascular endothelial growth factor-A in gastric carcinoma and their clinical significance.
Hou-long LONG ; Xiang-ming CHE ; Tie-qiang BI ; Hai-jun LI ; Jun-song LIU ; Duo-wei LI
Chinese Journal of Surgery 2012;50(9):839-842
OBJECTIVESTo study the expression of nicotinamide phosphoribosyl transferase (Nampt) and vascular endothelial growth factor-A (VEGF-A) in gastric carcinoma and investigate their correlations to clinicopathologic features and prognostic significance.
METHODSThe proteins of Nampt and VEGF-A in 68 specimens of gastric carcinoma and 59 specimens normal gastric tissue were detected by immunohistochemistry during January 2000 to December 2004, and the 68 patients were followed up.
RESULTSNampt protein was detected in the cytoplasm of both tissues, and Nampt in gastric carcinoma (13 ± 5) were significantly higher than that in normal gastric tissue (6 ± 3) (t = 7.46, P < 0.01). The expression of Nampt was correlated to invasive depth (F = 4.693, P = 0.034), lymph node metastasis (F = 4.027, P = 0.049), clinical TNM stage (F = 9.979, P = 0.002), but not to gender, age, tumor location, tumor size, differentiation (P > 0.05). The expression of Nampt is correlated with survival of patients that underwent surgical resection for gastric cancer. The survival rate of patients in negative of Nampt was very higher than that of the positive patients, and its co-expression with VEGF-A showed a trend towards poorer survival. The positive correlation was found between the expression of Nampt and VEGF-A in gastric carcinoma (r = 0.293, P = 0.015).
CONCLUSIONSThe expression of Nampt is positively correlated to that of VEGF-A in gastric carcinoma. The correlation between the expression of Nampt and VEGF-A in gastric carcinoma plays an important role cooperatively in carcinogenesis, development and metastasis of gastric carcinoma.
Adult ; Aged ; Aged, 80 and over ; Cytokines ; metabolism ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Nicotinamide Phosphoribosyltransferase ; metabolism ; Prognosis ; Stomach Neoplasms ; metabolism ; pathology ; Survival Rate ; Vascular Endothelial Growth Factor A ; metabolism
10.Dental caries in ancient Chinese in Xia Dynasty.
Wei WANG ; Xiang-long ZENG ; Wu LIU
Chinese Journal of Stomatology 2008;43(5):308-310
OBJECTIVETo study the ancient teeth helped us to acquaint with the evolution of oral diseases.
METHODSTeeth and jaws excavated in Erlitou, Henan province and Youyao, Shanxi province were observed clinically with care. The total specimens included 56 individuals (38 males and 18 females, including 894 permanent teeth and 86 dental arches). The dental caries were observed. The incidence of dental caries was calculated.
RESULTSThe rate of dental caries was 6.9% in 894 observed teeth. The prevalence was 39% among individuals.
CONCLUSIONSThe rate of dental caries in ancient Chinese in Xia Dynasty was lower than that in modem people. There were more caries in elderly people while no sex difference was found in Chinese in Xia Dynasty, which was similar to the situation in modern society. Cervical caries was more common than occlusal caries. The second molars and the third molars were frequently involved, which was different from the phenomenon nowadays.
Adult ; Archaeology ; Asian Continental Ancestry Group ; Dental Caries ; epidemiology ; Female ; Humans ; Male ; Prevalence