OBJECTIVETo study the effect of hypoxia on cofilin activation in intestinal epithelial cells and its relation with distribution of tight junction protein zonula occludens 1 (ZO-1).

METHODSThe human intestinal epithelial cell line Caco-2 was used to reproduce monolayer cells. The monolayer-cell specimens were divided into control group (no treatment), hypoxic group ( exposed to hypoxia), and normoxic group (exposed to normoxia) according to the random number table. Western blotting was used to detect the protein expressions of cofilin and phosphorylatedl cofilin (p-cofilin) of cells in normoxic group and hypoxic group exposed to normoxia or hypoxia for 1, 2, 6, 12, and 24 h and control group, with 9 samples in control group and 9 samples at each time point in the other two groups. The other monolayer-cell specimens were divided into hypoxic group (exposed to hypoxia) and control group (no treatment) according to the random number table. Cells in hypoxic group exposed to hypoxia for 1, 2, 6, 12, and 24 h and control group were obtained. Morphology and distribution of F-actin was observd with laser scanning confocal microscopy, the ratio of F-actin to G-actin was determined by fluorescence method, and distribution of ZO-l and cellular morphology were observed with laser scanning confocal microscopy. The sample number of last 3 experiments was respectively 3, 6, and 3 in both hypoxic group (at each time point) and control group. Data were processed with paired ttest, analysis of variance of repeated measurement, and LSD-t test.

RESULTSThe protein expressions of cofilin and p-cofilin of cells between normoxic group exposed to normoxia for 1 to 24 h and control group showed no significant changes (with values from -0.385 to 1.701, t(p-cofilin)values from 0. 040 to 1.538, P values above 0.05). There were no obvious differences in protein expressions of en filmn of cells between hypoxic group exposed to hypoxia for 1 to 24 h and control group ( with values from 1.032 to 2.390, P values above 0.05). Compared with that in control group, the protein expressions of p-cofilin of cells were greatly reduced in hypoxic group exposed to hypoxia for 1 to 24 h (with values from 4.563 to 22.678, P values below 0.01), especially exposed to hypoxia for 24 h. The protein expressions of cofilin of cells between normoxic group and hypoxic group at each time point were close ( with t values from -0.904 to 1.433, P values above 0.05). In hypoxic group, the protein expressions of p-cofilin of cells exposed to hypoxia for 1, 2, 6, 12, and 24 h were 0.87 +/- 08, 0.780 .05, 0.89 +/- 0.07, 0.68+0. 07, and 0.57 +/- 0.06, respectively, significantly lower than those in normoxic group (0.90 +/- 0.07, 0.97 +/- 0.06, 1.00 +/- 0.06, 1.00 +/- 0.05, and 0.99 +/- 0.05, with t values from 3.193 to 16.434, P values below 0.01). In control group, F-actin in the cytoplasm was abundant, most of it was in bunches. The trend of F-actin was disorderly in hypoxic group from being exposed to hypoxia for 1 h, shortened in length or even dissipated. The ratios of F-actin to G-actin of cells in hypoxic group exposed to hypoxia for 12 and 24 h (0.89 +/- 0.12 and 0.84 +/- 0.19) were obviously decreased as compared with that in control group (1. 00, with t values respectively 3. 622 and 3. 577, P values below 0.01). There were no obvious differences in the ratios of F-actin to G-actin of cells between hypoxic group exposed to hypoxia for 1, 2, and 6 h and control group ( with values from 0.447 to 1.526, P values above 0.05). In control group, cells were compact in arrangement, and ZO-1 was distributed continuously along the cytomnembrane. From being exposed to hypoxia for 2 h, cells became irregular in shape in hypoxic group. ZO-1 was distributed in discontinuous fashion along the cytomembrane with breakage in hypoxic group exposed to hypoxia for 24 h.

CONCLUSIONSHypoxia may cause the disorder of dynamic balance between F-actin and G-actin by inducing cofilin activation, which in turn leads to the changes in distribution of tight junction protein ZO-1 in intestinal epithelial cells.

Actin Depolymerizing Factors ; Actins ; Blotting, Western ; Caco-2 Cells ; drug effects ; physiology ; Epithelial Cells ; cytology ; drug effects ; Humans ; Hypoxia ; metabolism ; Intestinal Mucosa ; drug effects ; metabolism ; pathology ; Intestines ; Oxygen ; pharmacology ; Tight Junctions ; drug effects ; metabolism ; Zonula Occludens-1 Protein ; metabolism

Biomedical Research ; China ; Preventive Medicine ; Research Support as Topic

Biomedical Research ; China ; Preventive Medicine ; Research Support as Topic

Humans ; Hypertension, Pulmonary ; metabolism ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; metabolism

Objective To evaluate the efficacy and toxicity of weekly docetaxel and cisplatin in previously untreated patients with advanced non-small-cell lung carcinoma. Methods Between January 2002 and December 2003 ,34 patients with pathologically comfirmed advanced non-small-cell lung carcinoma who had not received treatment were enrolled. The mean age was under 66 years. The patients received intravenous infusions of docetaxel(25 mg/m2,dayl ,8,15) with dexamethasone premedication and cisplatin(25 mg/m2,dayl ,8,15) ,followed by a week of rest. The remedies which were less than 6 regimens lasted to disease progression or severe toxicity. Therapeutic effect was evaluated by CT scan every two courses . The patients were followed up for 24 months. Descriptive statistics and SPSSIO. 0 software were used to analyse the results. Results 34 patients finished 90 courses. The mean was 2. 6 courses. All patients were followed up. Two patients achieved complete responses, ten patients achieved partial responses, ten patients achieved stable disease. An objective response rate of 35. 29% (95% confidence interval 19. 25%-51. 33% )was obtained. Patients life quality was significantly improved. The median time to progression was 4. 1 months, and median overall survival was 11 months. The 1-year survival rate was 47. 06% , the 2-year survival rate was 11.76% . Toxicities were mild. Grade 3 to 4 neutropenia (11.76%), anemia (5.88%), hyponatremia (5.88%), alopecie (17.64%) and nausea/vomiting (5. 88% ) were observed. Conclusion Weekly Cisplatin plus docetaxel is an effective and well-tolerated regimen in chemo-naive patients with advanced NSCLC. Well-designed clinical trials should be conducted.

Objective To investigate the association between arsenic(+3 oxidation state) methyltransferase (AS3MT) genetic polymorphism and susceptibility to endemic arsenism.Methods Polymerase chain reactionrestriction fragment length polymorphism-single strand conformation polymorphism(PCR-RFLP-SSCP) technology was performed to detect mutations of AS3MT gene intron 8 and exon 9 in genome DNA of the 79 cases and 110 controls.PCR products with abnormal band forms were further sequenced to find the types and sites of mutation.Chi-square test and multivariate Logistic analyses were conducted.Results The incidence of the 9149 base mutation(A→C) in AS3MT gene intron 8(AS3MT-9149) in case group(19.0％,15/79) was lower than that in control group (23.6％,26/110).The incidence of the codon 287 mutation(ATG→AT/CG) in AS3MT gene exon 9(AS3MT-287)in case group(10.1％,8/79) was lower than that in control group (11.8％,13/110).However,statistical analysis indicated no significant difference in both mutations between two groups[AS3MT-9149:odds ratio(OR) =0.59,95％ confidence interval(CI):0.26-1.31,P =0.195; AS3MT-287:OR =0.85,95％ CI:0.32-230,P =0.751].Conclusions There are no significant association between the genetic polymorphisms of AS3MT-9149,AS3MT-287 and the susceptibility to endemic arsenism.Similarly,due to small sample amount,we can not exclude the possibility that these gene polymorphisms are related to susceptibility to endemic arsenism.

Objective To investigate rheumatic disease manifestations in patients with myelodysplastic syndromes and explore possible causes.Methods Nine myelodysplastic syndromes(MDS)inpatients with rheumatic disease manifestations were reviewed retrospectively.Results Nine patients with the diagnosis of MDS had active rheumatic disease manifestations with various degree of hypergammaglobulinemia and positive autoantibodies.Two patients had rheumatoid arthritis(RA)and RA+anti-phospholipid syndrome(APS),four systemic lupus erythematosus(SLE)and SLE+polymyositis(PM),one patients had adult onset Still's disease, one with acute arthritis and cutaneous vassulitis,one had lupus-like manifestations including polyserositis,al- buminuria,hypocomplementemia and positive Commb's test.Conclusion The rheumatic disease manifesta- tions in patients with myelodysplastic syndromes are characterized by various active rheumatic disease manifes- tation.Immunological abnormalities and ineffective hematopoiesis are persistent.Immunologic abnormalities in MDS such as impaired function of T and B cells,hypergammaglobulinemia,positive autoantibodies,may be the causes of various rheumatic manifestations.We suggest that there is a significant association between myeludys- plastic syndrome and rheumatic diseases.

Objective To establish a network laboratory for blood cell analysis and better calibrate haematology analyzers in local lab.Methods According to GB/T 15481《General requirements for the competence testing and calibration laboratories》(idt ISO/IEC 17025),we established a network laboratory providing traceability for blood cell analysis.Complete blood count was traced to Calibration Laboratory in NCCL;The secondary standard haematology analyzer with the same model and calibrator with same lot number were used for verification for a long period.Fresh blood from healthy people was used to calibrate haematology analyzers.Results Gradually we have improved our laboratory quality management system, precision as well as accuracy,which was satisfactory.The unified blood sample was adopted to calibrate different equipments in our hospital and showed consistence when compared with calibration analyzer.The correlation coefficient of all tests is more than 0.99.The relative deviation of WBC,RBC,HCT,HGB and PLT are within?7%,?3.5%,?4%,?3% and?15%,respectively.Conclusions Secondary standard systems provides good comparable results with calibration laboratory.Its tracing mode and quality control scheme could ensure the traceability and accuracy of completed blood count.Furthermore,using elective fresh blood from healthy people,the comparable results from different analyzers were achievable.

Objective To evaluate the feasibilities and advantages of different concentrations of sodium alginate (SA) solutions as a submucosal injection solution for endoscopic submucosal dissection (ESD). Methods In vitro study, different concentrations of sodium alginate solutions and normal saline were injected into submucosal of resected porcine esophagus and stomach respectively, then observe and measure the heights of each injection induced mucosal elevations, and their changes over time. In vivo study, the mimic ESD were conducted in healthy pigs to evaluate the mucosal elevation effect and other assistant effects of sodium alginate as a submucosal injection solution. Results The elevation heights of the experiment groups injected with SA solutions were much higher than the control group injected with normal saline. Specially, the elevation created by 1 % SA in porcine esophagus was significantly higher than that of normal saline (P < 0.01) and the elevation created by 3 % SA was significantly higher than that of normal saline in porcine stomach (P < 0.001). In the mimic ESD experiment, mucosal elevation with clear margin occurred immediately after injection with SA solution. And the durable submucosal fluid cushion created by SA protected deeper tissues while facilitating ESD procedure. Conclusion The elevation heights created by SA solutions were greater and more durable than that created by normal saline, which were crucial for ESD. The viscosity property enabled SA to form a stable protective cushion and prevent bleeding by squeezing tissue around the wound, which may decrease perforation and bleeding rate during ESD procedure. Therefore, sodium alginate can be an ideal clinical submucosal injection solution.

Objective To investigate the long-term anticoagulation starus and relation of INR and complica-tion,best anticoagulation range and clinical significance in patients after mechanical heart valve replacement. Meth-ods The data of blood clotting test series(containing PT,PT% ,PTR,INR,Fbg,APTT) for patients after 6 months of mechanical heart valve replacement were collected, then the cases were divided into 4 groups according to their INR levels( group Ⅰ : INR < 1.5; group Ⅱ : INR 1.5 ～ 2.0 ; group Ⅲ : INR 2.1 ～ 2.5 ; group Ⅳ : INR > 2.5 ). Results Group Ⅰ contained 28 person-times, with their dosage of warfarin for (3.61 ± 1.44 ) mg, INR 1.38 ± 0. 12, core-bral embolism occurred for 4 person-times,with a inception rate of 14%. Group Ⅱ contained 92 person-times,with their dosage of warfarin for (3.5±1.37)mg,INR 1.65±0. 14,hemorrhinia occurred for 2 person-times,with a in-ception rate of 2%. Group Ⅲ contained 80 person-times, with their dosage of warfarin for (3.18±1.63 )mg, INR 2.23±0.19 ,bematuria occurred for 2 person-times, bemorrhinia occurred for 2 person-times, with a total inception rate of 5% ; Group Ⅳ contained 16 person-times, with their dosage of warfarin for ( 2.32 ± 1. 23 ) mg, INR 2.80± 0.19, hemorrhinia occurred for 2 person-times and hemoptysis occurred for 1 person-times, with a total inception rate of 18.7%. The person-times in group Ⅱ and Ⅲ accounted for 79.6% and the complication rate was the lowest when INR was between 1.5-2.5. Conclusions INR levels between 1.5-2.5 is the most secure, which is the best antico-agu]ation range in our region. Since the anticoagulation strength dynamically changes, the long-term anticoagulation status in patients after mechanical heart valve replacement should be deeply concerned.