Adenoviridae ; genetics ; Cell Line ; Genetic Vectors ; Humans ; Nitric Oxide Synthase Type III ; genetics ; Virus Replication

Objective To prepare monoclonal antibody(McAb) against human tissue kallikrein (HK) and develop an ELISA kit allows for the in vitro quantitative determination of human tissue kallikrein in urine. Methods To generate a monoclonal antibody specific for TK, the synthetic TK peptide consisting of 12 amine acids(12P), was fused to keyhole limpet hemocyanin(KLH) and used for immunization. Using hybridoma screening, monoclonal secreting cell lines were identified and used to generate ascites in BALB/c mouse. Antibody was purified by affinity column chromatography. 12% SDS-PAGE and Western blot were used to visualize the purified antibody. This kit employs indirect competitive ELISA technique and BiotinAvidin System. 12P was fused to bovine serum albumin(BSA) and has been pre-coated onto a microplate at first. Standards and samples were added to the appropriate microplate wells with a biotin-conjugated McAb croplate well. A TMB substrate solution is added to each well. The enzyme-substrate reaction is terminating by the addition of a sulphuric acid solution and the color change is measured spectrotometrically at a wavelength of 450 nm. The concentration of tissue kallikrein in the samples is then determined by comparing the O.D. of the samples to the standard curve. Results 8 hybridoma cell lines secreting mAbs special to HK,SDS-PAGE and Western blot demonstrated successful preparing and purification of McAb( 100% ). The linearity of this ELISA kit is demonstrated(r =0. 990). The range of detection of the assay is 0.008 μg/ml to 0. 5 μg/ml. The assay remained stable, with no change in the values measured, over five cycles of freezing and thawing. Conclusion 8 McAbs against HK have been prepared successfully and possess high titer and specificity. The development of an ELISA kit for detecting HK can meet the needs of detection of HK in urine samples.

Objective To assess the agreement of different endoscopy grading or scoring systems for inflammatory bowel diseases(IBD)including ulcerative colitis(UC)and Crohn's disease(CD).Methods A standardized table was prepared based on the searches for endoscopic grading or scoring systems on Medline and Chinese Biomedical Database,the data of 80 patients with UC and 31 with CD.who underwent colonoscopy in Shanghai Renji hospital from June 2006 to February 2007,were evaluated with each system by two physicians independently.Data were analyzed with SPSS 13.0.Results Six endoscopic grading and scoring systems of UC and three of CD were included for evaluation.For the systems of UC and CD,Kendall's coefficients of concordance were 0.71(P<0.01)and 0.34(P<0.01),respectively.There was no significant differenee between every two systems for UC.Nonetheless.Spearman's correlation coefficient between Chinese Grading System of Crohn's Disease(CGSCD)and Crohn's Disease Endoscopic Index of Severity(CDEIS)was 0.32(P=0.08).Significant differences in frequencies were detected in endoscopic systems for UC by Kruskal Wallis test(P<0.01).Conclusion There is satisfactory concordance among the endoscopic grading and scoring systems of UC,while CGSCD needs further improvement.Furthermore,Jeroen elassifieation inclines to severe category,while modified Baron scale tends to be a mild one.

Humans ; Occupational Diseases ; diagnosis

BACKGROUND:Col agen and silk fibroin materials for construction of spinal cord scaffolds have been proven to repair or partial y repair damaged spinal cord nerve function. OBJECTIVE:To introduce partial characteristics of the col agen and silk fibroin and to review the recent progress and application as scaffolds in spinal cord tissue engineering. METHODS:A computer-based search of CNKI and PubMed databases (2003-01/2012-10) was performed for articles addressing the application of col agen and silk fibroin scaffolds in spinal cord injury with the keywords of“col agen, silk fibroin, scaffold, spinal cord injury”in Chinese and English, respectively. RESULTS AND CONCLUSION:Col agen has low antigenicity, good biocompatibility and biodegradability. Col agen and its degradation products can cause no inflammatory reactions in the body, but have the disadvantages of rapid degradation and poor mechanical properties. Silk fibroin has good biocompatibility and excellent mechanical properties, but its degradation is slow. The col agen and silk fibroin are compounded using an electrostatic spinning technology to improve the physical properties of the material on the basis of maintaining good biocompatibility. At present, fibroin or col agen materials in terms of nervous system repair have been studied, laying some foundation for spinal cord tissue engineering. Considering the similar characteristics and mechanics performance to the spinal cord tissue, col agen/silk fibroin composite materials are expected to become the ideal scaffold materials for spinal cord tissue engineering.

Herbicides ; poisoning ; Humans ; Paraquat ; poisoning ; Poisoning ; metabolism ; pathology ; therapy

BACKGROUND:Neural stem cel s with self-proliferation and differentiation potential are the ideal seed cel s for central nervous tissue engineering. Although col agen and silk fibroin as biological scaffold materials have been widely used, both of them used alone have certain shortcomings. Is it possible to combine the two materials to build a novel neural tissue-engineered scaffold? What is the effect of this novel scaffold on the growth and differentiation of neural stem cel s? OBJECTIVE:To observe the growth and differentiation of neural stem cel s seeded onto the novel composite scaffold. METHODS:The rat embryonic neural stem cells were inoculated onto new composite scaffolds, and then, their growth and differentiation were observed by light microscopy and scanning electron microscopy. Neural stem cells were cultured in conventional suspension culture as control group. Cell counting kit-8 assay was used to detect viability of neural stem cells in the two groups. Three-dimensional composite scaffolds carrying neural stem cells were slic ed into paraffin sections to observe the growth and differentiation of neural stem cells by hematoxylin-eosin staining and immunofluorescence staining. RESULTS AND CONCLUSION:Neural stem cel s cultured on the new composite scaffold grew and differentiated wel , and interconnected synapses were observed. Cel counting kit-8 assay showed that neural stem cel s on the scaffold grew wel , and the cel viability was significantly higher in the composite scaffold group than that in the control group (P<0.05). Hematoxylin-eosin staining and immunofluorescence staining of paraffin sections further provided evidence for good growth and differentiation of neural stem cel s on the scaffold. These results indicate that the novel composite scaffold with good biocompatibility benefits the growth and differentiation of neural stem cel s, promising a favorable application prospect.

By updating their concepts of education,carrying out the reform of teaching content,teaching methods and means,strengthening the construction of teaching materials,creating a high standard teaching team,the author aims to implement the construction of quality course of clinical testing equipment and comprehensively improve the teaching quality.

Objective To investigate the clinical features of Crohn's disease(CD)and ulcerative colitis(UC)according to the Montreal classification.nethods The clinical data of 110 cases of CD or UC were reviewed.The age at diagnosis.location and clinical behavior were assessed with the Montreal criteria.Results CD patients diagnosed at an age younger than 16 years were rare(3.6%),the majority of the CD patients was diagnosed at 17-40 years old(65.5%).Although ileocolon lesions were most common in the patients diagnosed at 17-40 years old(37.3%),yet ileum lesions were a little more than those of other parts of digestive tract in the patients diagnosed after 40 years old(14.5%),the difference was not significant(P=0.054).Stricture frequently occurred(50.4%),especially when the lesions were located at ileum or ileocolon.Perforation rarely happened(5.3%).There was no significant difference between different location groups for clinical behaviors(P=0.096).The incidence of stricture or perforation was almost same among different age groups(P=0.984).UC patients mostly presented with mild or moderate symptoms even in the group with extensive lesion.UC patients with severe symptoms were rare(6.8%).There was no significant difference in severity between the groups with different extent of lesion (P=0.056).Conclusions The majority of CD patients was diagnosed at 17-40 years old.Stricture is much more than perforation,penetrating,occurring mostly at ileum.UC patients mostly present mild or moderate symptoms,no matter how extensive the lesion is.The extent of the lesion detected with coloscopic examination is not correlated to the severity of clinical manifestations.