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.

Air Pollutants, Occupational ; chemistry ; Indium ; analysis ; Mass Spectrometry ; methods ; Occupational Exposure ; Spectrum Analysis ; methods ; Workplace

Chromatography, Gas ; Humans ; Hydrocarbons, Brominated ; urine

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.

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.

Cross-Sectional Studies ; Dust ; Humans ; Occupational Exposure ; adverse effects ; Pneumoconiosis ; epidemiology ; Prevalence

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

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

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.

OBJECTIVETo evaluate the clinicopathologic features of gastrointestinal stromal tumor (GIST) with synchronous carcinoma and the treatment principle.

METHODSNineteen cases of GIST with synchronous carcinoma were collected from 113 cases of GIST from 2002 to 2008. The clinicopathologic features were studied and the expression of CD117, CD34, smooth muscle actin and S-100 protein were detected by immunohistochemistry using EliVision method. The expression of proliferation marker Ki-67 was also studied. GIST with synchronous carcinoma and those without carcinoma were compared.

RESULTSNineteen cases (16.8%) of GIST with synchronous carcinoma were found, including 11 males and 8 females (male to female ratio 1.38: 1.00). The age of the patients ranged from 43 to 66 years (median age 57 years). Five of 19 cases were located in the inferior segment of esophagus and 14 were in the gastric wall. The diameter ranged from 0.6 to 3.8 cm [mean (1.91 ± 0.92) cm]. Three of 19 cases showed low grade dysplasia, and there was no dysplasia in the remaining 16 cases. The number of mitosis ranged from 0 to 4/50 HPF [mean (0.74 ± 1.07)/50 HPF]. The Ki-67 proliferative index (number of Ki-67 positive cell/HPF) ranged from 0 to 7.72% [mean (2.51 ± 2.20)%]. The synchronous carcinomas included two esophageal carcinomas and 17 gastric cancers.In contrast, patients of GIST without carcinoma included 52 males and 42 females (male to female ratio 1.24: 1.00). The age of patients ranged from 43 to 71 years (median age 55 years). Seventy-nine of the 94 cases were located in the stomach, 10 were in the intestine and 5 were in the esophagus. The diameter ranged from 2.4 to 15.5 cm [mean (5.42 ± 6.17) cm].Seventy-nine of the 94 cases showed variable degrees of dysplasia, and 12 cases were of high malignant potential. The number of mitosis ranged from 0 to 53/50 HPF [average (3.78 ± 10.22)/50 HPF]. The Ki-67 proliferative index ranged from 0 to 37.54% [mean (6.78 ± 12.45)%]. Comparing these two groups, the male to female ratio of GIST with synchronous carcinoma was higher than that of GIST without carcinoma. The average diameter of GIST with synchronous carcinoma was smaller than of those without carcinoma. The number of mitosis and Ki-67 proliferative index of GIST with synchronous carcinoma were significantly lower than those without carcinoma (t' = 2.809, P < 0.05; t' = 3.095, P < 0.05, respectively).

CONCLUSIONSSixteen point eight percent of GIST may be associated with synchronous carcinoma. There are no special clinical symptoms in most of GIST with synchronous carcinoma, as these GIST are usually incidental findings. The Ki-67 proliferative index of GIST with synchronous carcinoma is significantly lower than that of GIST without synchronous carcinoma. Most GIST with synchronous carcinoma can be treated by the standard treatment for the accompanying carcinoma, and do not require specific additional treatments.

Adenocarcinoma ; metabolism ; pathology ; therapy ; Adenocarcinoma, Mucinous ; metabolism ; pathology ; therapy ; Adult ; Aged ; Antigens, CD34 ; metabolism ; Carcinoma, Signet Ring Cell ; metabolism ; pathology ; therapy ; Carcinoma, Squamous Cell ; metabolism ; pathology ; therapy ; Chemotherapy, Adjuvant ; Esophageal Neoplasms ; metabolism ; pathology ; therapy ; Esophagectomy ; Female ; Follow-Up Studies ; Gastrectomy ; Gastrointestinal Stromal Tumors ; metabolism ; pathology ; therapy ; Humans ; Ki-67 Antigen ; metabolism ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasms, Multiple Primary ; metabolism ; pathology ; therapy ; Proto-Oncogene Proteins c-kit ; metabolism ; Radiotherapy, Adjuvant ; Stomach Neoplasms ; metabolism ; pathology ; therapy