1.Influence of surgery on the regulatory T cell levels in peripheral blood for patients with cavitary tuberculosis
Yin′e WU ; Wenguang PENG ; Qinghua WU ; Gaozhe ZHENG ; Shiyang LIN
Chinese Journal of Primary Medicine and Pharmacy 2014;(9):1292-1293
Objective To analyze the regulatory T cell ( CD4 +CD25 +FoxP3 +T) levels in peripheral blood of patients with cavitary tuberculosis before and after surgery .Methods We compared the proportion of CD4 +CD25high T cells and CD4 +CD25 +FoxP3 +T cells in 13 patients with cavity tuberculosis pre-and post-operatively and in 10 healthy control subjects with flow cytometry .Results (1) The proportion of CD4 +CD25high T cells and CD4 +CD25 +FoxP3 +T cells were significantly higher in patients with cavity tuberculosis and at 1-month postoperatively than that in healthy controls(P<0.01).The proportion of CD4 +CD25high T cells and CD4 +CD25 +FoxP3 +T cells had no statistical significance between the patients with cavity tuberculosis at 6-months postoperatively and healthy controls (P>0.05).(2)The proportion of CD4 +CD25high T cells and CD4 +CD25 +FoxP3 +T cells decreased significantly af-ter 6 months surgery than preoperatively in patients with cavity tuberculosis (P<0.01).(3)Pre-and post-operative proportions of CD4 +CD25high T cells and CD4 +CD25 +FoxP3 +T cells showed a positive correlation ( r=0.878,P<0.01 ) .Conclusion The proportion of the circulating T regulatory cells increases in patients with cavity tuberculosis , and decreases after surgery .Infection with M.tuberculosis may induce regulatory T cell-surface molecular changes.
4.Experimental Study of Vascular Tissue Engineering
Tai YIN ; Yingxin XU ; Shihe WU ; Jinjing WANG ; Lingling E ; Rong LI ; Ruixin LI ; Xizheng ZHANG
Chinese Journal of Rehabilitation Theory and Practice 2007;13(6):526-527
Objective To obtain the experimental data of vascular tissue engineering.MethodsThe vascular endothelial cells (VEC) and vascular smooth muscle cells (VSMCs) were acquired and cultured, and then seeded on vascular tissue engineering materials. The porous gelatin-chitosan scaffold with VSMCs was subcutaneously implanted, followed by the observation of the cell growth ten days later.ResultsThe two kinds of cells were successfully cultured and their morpholoical and immunohistochemical characteristics were consistent with vascular endothelial and VSMCs respectively. The VSMCs could grow extensively on the scaffold after the in vivo implantation. The scaffold were wrapped by the fibrous tissue ten days later after the in vitro implantation of VSMCs. The seed cells grew in the scaffold, and the vessel cavity seen in the center of the scaffold, was quite different from the normal vessel structure.ConclusionIt is feasible to implant the VSMCs with fibrin gels into the living body. The vessels reconstructed, though different from the normal structure, is similar to the embryo of the vessels.
5.Elementary study on construction of engineered blood vessel using chitosan-fibrin gel as scaffold
Yingxin XU ; Tai YIN ; Ruixin LI ; Shihe WU ; Jinjing WANG ; Lingling E ; Xizheng ZHANG ; Rong LI
Chinese Journal of Rehabilitation Theory and Practice 2005;11(8):623-625
ObjectiveTo investigate the feasibility of construction of engineered blood vessel using chitosan tube and fibrin gel as scaffold.MethodsVascular endothelial cells and smooth muscle cells were harvested from aortas of a rat, respectively. After expansion in vitro, vascular endothelial cells were seeded onto the inner surface of chitosan tube and smooth muscle cells mixed with fibrin gel seeded onto outer surface of the scaffold to construct engineered blood vessels. Inverted microscope, immunohistochemical staining and scanning electronic microscope were used to evaluate the construct.ResultsVascular endothelial cells formed monolayer and covered the inner surface of chitosan tube. Smooth muscle cells survived in the fibrin gel and grew in a 3-dimensional manner. ConclusionChitosan-fibrin gel may be potentially used as scaffold of engineered blood vessels.
6.Significance of CT in diagnosis of chronic suppurative otitis media.
Lin-e WANG ; Yi-feng GU ; Ya-qin WU ; Qi-xin ZHUANG ; Yan LIN ; Shan-kai YIN
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2007;42(7):494-498
OBJECTIVETo investigate the methods of preoperative diagnosis and differentiation of different pathological tissue in middle ear and mastoid.
METHODSThe temporal bone lamellar CT findings in 106 patients with chronic suppurative otitis media (including cholesteatoma) were retrospectively analyzed. The CT value of pathological tissue were measured for 183 times and were compared with the surgical findings and postoperative pathological findings to definitude the CT value range of different pathological tissue. Sixty patients taken from 106 patients at random were analyzed and made the diagnosis again by the same doctor team according to the CT value of the different pathological tissue and surrounding histoclasia resulted by pathological tissue. The diagnose accordance rate was compared with the routine diagnose report from radiology department. The predetective diagnosis was made in 10 patients with chronic suppurative otitis media according to clinical manifestation (pathological changes of tympanic membrane, nature of otorrhea, character of hearing), temporal bone lamellar CT finding (CT value of pathological tissue, surrounding histoclasia) to validate the value of this study for preoperative diagnosis and differentiation of different pathological tissue in middle ear and mastoid.
RESULTSThe CT value of cholesteatoma, granulation tissue, cholesteatoma combined with granulation tissue, effusion, calcified tissue, thickened and polypoid membrane was respectively (46.6 +/- 10.3) Hu, (26.6 +/-7.4) Hu, (42.1 +/- 11.4) Hu, (- 24.6 +/- 9.2) Hu, (223.6 +/- 63.7) Hu, (23.8 +/- 8.5) Hu. The diagnose accordance rate in 60 patients who were analyzed and made diagnosis again according to the CT value of the different pathological tissue and surrounding histoclasia resulted by pathological tissue raised from 68. 3% to 81.7% ( P < 0.05) . The predetective diagnose accordance rate reached at 90% according to clinical manifestation, temporal bone lamellar CT.
CONCLUSIONSIt was not reliable to diagnose and differentially diagnose different pathological tissue in middle ear and mastoid only by the CT value, however, the CT value could still be considered to be a very significant information. The accurate rates of diagnosis and differentiation of different pathological tissue in middle ear and mastoid obviously raised by synthetically analyzing various kinds of pathological tissues in middle ear and mastoid according to clinical manifestation, temporal bone lamellar CT finding.
Adolescent ; Adult ; Aged ; Child ; Chronic Disease ; Female ; Humans ; Male ; Mastoid ; diagnostic imaging ; Middle Aged ; Otitis Media, Suppurative ; diagnostic imaging ; Retrospective Studies ; Temporal Bone ; diagnostic imaging ; Tomography, X-Ray Computed ; Young Adult
8.Ultrastructural characteristics of SARS associated virus in infected cells.
Cui-e WANG ; Yu-chuan LI ; Xiao-hong WU ; Jun-tian CAO ; Ge YAN ; Jin-feng LI ; Bing-yin SI ; Man YU ; E-de QIN ; Qing-yu ZHU
Chinese Journal of Pathology 2003;32(3):209-211
OBJECTIVEElectron microscopical study of infected cells to identify the pathogenic agent of SARS.
METHODSVero E6 cells infected with lung autopsy samples or nasopharyngeal swabs from SARS patients of Beijing and Guangzhou were inoculated. The supernatant and cultured cells exhibiting identifiable cytopathic effect (CPE) were prepared for electron microscopic study.
RESULTSExamination of CPE cells on thin-section revealed characteristic coronavirus particles within the cisternae of endoplasmic reticulum, Golgi apparatus, vesicles and extracellular space. They were mainly spherical or oval in shape, annular or dense, about 80 nm in diameter. Negative-stain electron microscopy identified coronavirus particles in culture supernatant, 80 - 120 nm in diameter, with club-shaped surface projections. Elongated, rod-, kidney- or other irregular shaped virons with the size of 100 - 200 nm by 60 - 90 nm were also found in the cultured cells infected with the lung samples from the Guangdong patients. Infectious virons entered cells by endocytosis or membrane fusion and released through a budding process.
CONCLUSIONThese data indicate a novel coronavirus as the causative agent of SARS. Most viral particles showed typical characteristics of coronavirus. The potential role of special shape viruses is expected to be further investigated.
Animals ; Cercopithecus aethiops ; Humans ; Microscopy, Electron ; SARS Virus ; ultrastructure ; Severe Acute Respiratory Syndrome ; virology ; Vero Cells
9.CT-guided thoracic biopsy: evaluating diagnostic yield and complications.
Stanley E K LOH ; Donald D F WU ; Sudhakar K VENKATESH ; Cheng Kang ONG ; Eugene LIU ; Kar Yin SETO ; Anil GOPINATHAN ; Lenny K A TAN
Annals of the Academy of Medicine, Singapore 2013;42(6):285-290
INTRODUCTIONThis study retrospectively evaluated CT-guided thoracic biopsies for diagnostic yield, accuracy and complications.
MATERIALS AND METHODSA retrospective analysis of 384 patients (mean age 62.7 years; male/female = 251/133) who underwent 399 CT-guided thoracic biopsies were performed for evaluating diagnostic yield, accuracy and complications. Correlations between patients age, procedure factors (biopsy-needle size, number of passes, lesion-size, lesion-depth and traversed lung-length) and complications such as pneumothorax, haemothorax and haemoptysis were evaluated. A comparison between fine needle aspiration (FNA) group and core ± FNA group for diagnostic yield and complications was also performed.
RESULTSFNA was performed in 349 patients and core ± FNA in 50 patients. The biopsy samples were adequate in 91.9% and the diagnostic accuracy for malignant lesions was 96.8% with 95.7% sensitivity and 100% specificity. Pneumothorax (detected on CT) occurred in 139 cases (34.8%) and only 12 (3.0%) required insertion of an intercostals drain. Mild haemoptysis occurred in 13 patients (3.2%) and small haemothoraces in 2 patients. Pneumothorax occurrence was significantly associated with the traversed lung-length (>3mm), lesion-size (≤33 mm) and lesion-depth (≥60mm) (P <0.05). Haemoptysis occurrence was also significantly associated with traversed lunglength (>3mm) and lesion-size (≤33 mm) (P <0.05). There was no significant difference between diagnostic yield and complication rate between FNA and core ± FNA groups.
CONCLUSIONCT-guided thoracic biopsy is a safe procedure with high diagnostic yield and low risk of significant complications. Traversed lung-length and smaller lesion size are associated with occurrence of pneumothorax and haemoptysis.
Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Image-Guided Biopsy ; adverse effects ; methods ; Lung Neoplasms ; diagnosis ; Male ; Middle Aged ; Postoperative Complications ; Reproducibility of Results ; Retrospective Studies ; Tomography, X-Ray Computed
10.Construction of a lentiviral vector carrying short?hairpin RNA targeting PAX6 and its effect on proliferation of glioma U251 cells in vitro.
Xiao-Hong LIAO ; Wei-Lan YIN ; Fang WANG ; Li-Xiang WU ; Bai-Sheng HUANG
Journal of Southern Medical University 2017;37(12):1603-1608
OBJECTIVETo construct a lentiviral vector for delivering short hairpin RNA (shRNA) targeting PAX6 and investigate its effect on the proliferation of glioma U251 cells in vitro.
METHODSTwo small interfering RNA sequences targeting PAX6 gene were designed based on the reported sequence of PAX6 and annealed to form a double?stranded chain, which was inserted into a lentiviral vector to construct the recombinant lentiviral vector shRNA?PAX6. The recombinant vector was infected into U251 cells, and the expression of PAX6 mRNA and protein in the cells was detected by real?time PCR and Western blotting, respectively. The changes in the proliferation of U251 cells after the infection was assessed using MTT assay.
RESULTSDouble enzyme digestion of the lentiviral vector pLKD?CMV?G&NR?U6?shRNA yielded an 8208?bp fragment, and colony PCR and sequencing analysis confirmed successful construction of the lentiviral vector shRNA?PAX6. Infection of the cells with shRNA?PAX6 caused a significant reduction of the expressions of PAX6 mRNA and protein (P<0.05) and resulted in obviously increased proliferation of U251 cells (P<0.05).
CONCLUSIONWe successfully constructed the recombinant vector shRNA?PAX6 for silencing PAX6 gene. PAX6 gene silencing results in increased proliferation of U251 cells in vitro.