The effects of tetramethylpyrazine ( TMPZ ) on the contraction of isolated guinea pig trachea induced by leukotrienes,histamine,prostagladin F2a , acetylcholine were observed. It was found that TMPZ exerts an inhibitory effect on the contraction induced by these asthmogenic mediators. It is non-competitive antagonist.

In this study, the effect of prophylactic anti-inflammation on the development of smoke-induced emphysema was investigated. Young male guinea-pigs aged 1.5-2 months (weighing 198.3+/-26.9 g) were randomly divided into 4 groups: group A (cigarette smoke exposure only), group B (cigarette smoke exposure plus pentoxifylline-rich (PTX, 10 mg/d) forage feeding), group C (cigarette smoke exposure plus intermittent cortical steroid injection (Triamcinolone acetonide, 3 mg, i.m., every three weeks) and control group (group D: animals with sham smoke exposure, raised under the same conditions). Animals in group A, B and C were exposed to smoke of cigarettes for 1 to 1.5 h twice a day, 5 days a week. All animals were killed at the 16th week and followed by morphometrical analysis of the midsagittal sectioned lung slices. Smoke exposure of 16 weeks resulted in visible emphysematous development in Group A but not in Group B and C. It was evidenced by the indicator of air-space size, mean linear intercept (Lm): 120.6+/-16.0 microm in Group A; 89.8+/-9.2 microm in Group B and 102.4+/-17.7 microm in Group C. The average Lm in either group B or group C was shorter than that in Group A (ANOVA and Newman-Keuls test, F=8.80, P=0.0002) but comparable to that (94.8+/-13.2 microm) in group D (P>0.05). It is concluded that long-term prophylactic anti-inflammation inhibits pulmonary emphysema induced by cigarette smoking in the guinea pigs.
Anti-Inflammatory Agents/*pharmacology ; Pentoxifylline/pharmacology ; Pulmonary Emphysema/etiology ; Pulmonary Emphysema/pathology ; Pulmonary Emphysema/*prevention & control ; Random Allocation ; Smoking/*adverse effects ; Triamcinolone Acetonide/*pharmacology

The effects of exogenous p16(ink4a) gene on biological behaviors of human lung cancer cell line with homozygous deletion of p16(ink4a) gene were investigated. Exogenous p16(ink4a) gene was transfected by lipofectin into human lung cell line A549, in which p16(ink4a) gene was homozygously deleted. The expression of p16(ink4a) mRNA and protein was detected by RT-PCR and immunocyto-chemistry, respectively. The changes in the behaviors of the transfected cell lines in vitro and in vivo were observed. In the transfected cell line A549, the exogenous p16(ink4a) gene could be stably expressed. The growth of A549 cells transfected with p16(ink4a) gene was obviously slowed down. Flow cytometry revealed that transfection of the exogenous p16(ink4a) gene resulted in A549 cell lines arrest in G1 phase of cell cycle. The tumorigenicity of these transfected cells in nude mice could be inhibited, and the tumor growth of nude mice was significantly suppressed. It was concluded that exogenous p16(ink4a) gene may be stably expressed in human lung cancer cell line A549. The expression of the introduced p16(ink4a) could block lung cancer cells to entry into S phase of cell cycle and inhibit tumor malignant growth both in vitro and in vivo.

To examine the relationship between the levels of the serum vascular endothelial growth factor (VEGF) and the micrometastasis of peripheral blood in patients with non-small cell lung cancer (NSCLC), 108 NSCLC patients, including 40 patients with benign lung diseases and 30 healthy controls, were investigated. The serum VEGF levels were detected by ELISA and CK19 mRNA in peripheral blood by reverse transcriptase-polymerase chain reaction (RT-PCR). In NSCLC group, the serum VEGF levels and the positive rate of CK19 mRNA in peripheral blood were 479.8+/-268.5 pg/mL and 66.7%, which were significantly higher than those of the other two groups respectively (P<0.01), and both of them were increased significantly with the progression of clinical stage of the tumors (P<0.01). Serum VEGF levels as well as the positive rate of CK19 mRNA in different pathological types of lung cancer had no significant differences (P>0.05). Serum VEGF levels in the patients positive for CK19 mRNA was 561.7+/-325.6 pg/mL. It is significantly higher than that in the negative patients (P<0.01). There existed a significant correlation between serum VEGF levels and expression of CK19 mRNA in peripheral blood in NSCLC patients (P<0.001). The detection of serum VEGF levels and CK19 mRNA in peripheral blood is helpful in judging the condition and the prognosis of NSCLC patients, and serum VEGF levels and CK19 mRNA are independent of the pathological types of lung cancer. The micrometastasis in peripheral blood of NSCLC patients is significantly associated with serum VEGF levels.

Objective To test the reliability and validity of ICF Core Sets for Chinese COPD patients.Methods Fifty-two COPD patients were measured with ICF Core Sets for COPD patients and SF-36. For reliability test, the internal consistency was analyzed and expressed by Cronbach α coefficients and split-half reliability. For va-lidity test, the content validity and criterion validity were analyzed and expressed with Spearman rank correlation coef-ficients. Results For body function, body structure and activity and participation, there were good internal consis-tency (Cronbach α coefficients 0. 698～0.957). For environmental factors Crnnbach α coefficient and split-half reli-ability did not exist. Most items of body function, activity and participation and body structure possessed good content validity. There was concurrent validity for ICF components of body function, body structure and activity and participa-tion with SF-36, FEV1/FVC, COPD grade and health self-assessment. The environmental factors demonstrated poor reliability and validity. Conclusion The ICF Core Sets for COPD patients showed good reliability and validity. It is a good comprehensive functional measurement scale for COPD patients, but it is necessary to test the generality of this result, and some items need to be adjusted.

AIM:To investigate the effect of Polo-like kinase-1(Plk1) depletion on cell cycle progression and cell growth in lung cancer cells.METHODS:A recombinant plasmid containing antisense RNA targeting Plk1(pcDNA3-Plk1) was transfected into A549 cells by lipofectine.RT-PCR and Western blotting were used to examine Plk1 gene expression.Cell proliferation was evaluated by cell counting and BrdU labeling.Cell cycle distribution and apoptosis were examined by flow cytometry.Inhibition rate(IR) of vinorebline(NVB) was determined by MTT assay.RESULTS:After transfected with pcDNA3-Plk1 into A549 cells,the expression levels of Plk1 mRNA and protein were greatly decreased.Abnormal morphological changes of cells and growth inhibition were observed in pcDNA3-Plk1 transfected cells.The BrdU labeling index was significantly lower than that in control group(P

AIM: To localize the expression of forkhead/winged helix transcription factor(FOXP3) gene in different types of pathological lung tissues and explore its biological role in pathogenesis of human lung cancer.METHODS: By using RT-PCR and Western blotting,the expressions of FOXP3 mRNA and related protein in 153 samples including lung cancer(n=63),lung benign lesion(n=45) and normal lung tissues(n=45) were analyzed.RESULTS: The positive expressions of FOXP3 mRNA and its protein were observed in lung cancer and in benign lesion lung tissue samples with significant difference(P0.05).CONCLUSION: FOXP3 is a biomarker of CD4+CD25+ regulatory T cell.They are expressed both in lung cancer and benign lesion lung tissues,but not in normal lung tissue.The expression of FOXP3 is more intensive in cancer tissues than that in benign lesions.

To compare the difference in tumor immunity and autoimmunity elicited by adenovirus (Ad) encoding human or murine tyrosinase-related protein 2 (AdhTRP2 or AdmTRP2), and to find the most effective way to induce immunity by AdhTRP2 or AdmTRP2, C57BL/6 mice were immunized with AdhTRP2 or AdmTRP2 intramuscularly at different doses of 10(5), 10(6), 10(7) and 10(8) separately (10 mice for each dose). Two weeks after the immunization, in vivo CTL assay and intracellular staining (ICS) of IFN-gamma were carried out to analyze the dose-effect relationship. Tumor growth and vitiligo (as an sign of autoimmunity) were observed until 3 months after challenge with 10(5) B16F10 tumor cells. The results showed that Ad encoding AdmTrp2 induced weak tumor immune response. Similar immunization with AdhTrp-2 elicited stronger protective immunity. CTL activity and IFN-gamma-produced CD8+T cells were directly proportional to dose of AdhTrp2 or AdmTrp2. Moreover, AdhTrp2 group showed tumor rejection in 100% of challenged mice till the end of 3rd month while 60% of mice immunized with AdmTrp2 were protected against tumor. In the whole process of this experiment, no vitiligo was observed in mice immunized either with AdhTrp2 or AdmTrp2. It is concluded that anti-melanoma responses induced by genetic vaccination expressing xenoantigens breaks immune tolerance effectively and is able to elicit strong antigen-specific cytotoxic T cell response without vitiligo.
Adenoviridae/metabolism ; Antineoplastic Agents/*pharmacology ; Cell Line, Tumor ; Cytokines/metabolism ; Immune System ; Immune Tolerance ; Interferon-gamma/metabolism ; Intramolecular Oxidoreductases/*biosynthesis ; Intramolecular Oxidoreductases/*genetics ; Mice, Inbred C57BL ; T-Lymphocytes, Cytotoxic/*metabolism ; Vitiligo/*metabolism

AIM: To investigate whether RNA interference (RNAi) induced by small interference RNA (siRNA) could suppress Polo- like kinase- 1 (Plk 1 ) expression and its effects in A549 cells. METHODS: A recombinant plasmid containing siRNA targeting Plk1 ( psiRNA - hH1 - Plk1 ) was transfected into A549 cells with Lipofectamine 2000.Expressions of Plk1, cyclin B1 and p53 protein were detected by Western blotting. Cell proliferation was evaluated by direct cell counting, while cell cycle and apoptosis were examined by flow cytometry, and expression of α - tubulin was detected by immunofluorescence. RESULTS: The results demonstrated that sequence specific siRNA targeting Plk1 was capable of suppressing Plk1 expression, and reflecting in lower kinase activity in A549 cells. The level of Plk1 protein was reduced by at least 70％ after 48 h of psiRNA - hH1 - Plk1 treatment relative to controls. Expressions of cyclin B1 and p53 were increased greatly after Plk1 depletion, and cells showed absence of microtubule polymerization and spindle abnormalities in staining for α -tubulin. Growth inhibition, G2/M arrest and apoptosis were observed in psiRNA -hH1 -Plk1 transfected group. CONCLUSION: All these data suggest that siRNA targeted against human Plk1 may be a valuable tool in cancer therapy.

To investigate the prognostic effects of integrated traditional Chinese and Western medicine therapy without antibiotics in treatment of patients with severe acute pancreatitis (SAP).