Objective To investigate the medical effect of the ethanol extract of Acorus gramineus Sol.on arthritis of mice induced by collagen-Ⅱ,and explore the potential pharmacological mechanisms.Methods Arthritis mouse model was established by injection of admixture containing type Ⅱ collagen and Freund's complete adjuvant (FCA) in male BALB/c mice.Mice were divided into five groups:the normal control group (0.9% of sodium chloride solution),the model control group (0.9% of sodium chloride solution),tripterygium group (15 μg·kg-1 of tripterygium tablets), the high-dose of extract of Acorus gramineus Sol.group (60 mg·kg-1 extract of Acorus gramineus Sol.) and the low-dose of extract of Acorus gramineus Sol.group (15 mg·kg-1 extract of Acorus gramineus Sol.).Each group was administered once a day,lasting 21 days.During the experiment,ankles of all mice were measured at predetermined time.At the end of the experiment,blood of the mice was exsanguinated and centrifuged to get serum for measuring the levels of IL-1β,RF and TNF-α.Spleens of mice were dissected and weighed to calculate the spleen index.All arthritis ankles were dissected to make tissue section,and observed under microscope.Results Compared with the model control group,the perimeter of ankle joints of the high-dose of extract of Acorus gramineus Sol.group significantly changed 6 days after administration (P<0.05);That of the low-dose of extract of Acorus gramineus Sol.group significantly changed 12 days after administration (P<0.05);That of tripterygium group significantly changed 9 days after administration (P<0.05).As compared with the normal control group, the spleen index of the model control group was significantly different (P<0.01).As compared with the model control group,the spleen index of tripterygium group,high-dose and low-dose of extract of Acorus gramineus Sol.groups were significantly different (P<0.05).As compared with the normal control group,levels of IL-1β,RF and TNF-α of the model control group were significantly different (all P<0.01).As compared with the model control group,levels of IL-1β,RF and TNF-α of tripterygium group,high-dose and low-dose of extract of Acorus gramineus Sol.groups were significantly decreased.Conclusion Ethanol extracts of Acorus gramineus Sol.have significant therapeutic effect on arthritis mice.The anti-arthritic mechanism is associated with its ability to regulate levels of IL-1β,RF and TNF-α.

In order to improve the knowledge of clinicians in hypoxia of COPD during the night,the physiological changes of arterial oxygen and carbon dioxide during sleep in healthy population were investigated at first,and the features and state of oxygen deficiency during sleep in patients with COPD were analyzed.Furthermore,the possible mechanism,effect,predictor,diagnosis and therapy were explored.

The effect of cigarette smoke extract (CSE) on the proliferation of human airway epithelial cells and the possible mechanism was studied. After airway epithelial cells were treated with different concentrations of CSE for 24 h, the cell proliferation was measured by MTT and the distribution of different cell cycles by flow cytometry. The FAK expression level was detected by Western blot and the degree of tyrosine phosphorylation by immunoprecipitation. The results showed that CSE could inhibit the proliferation of human airway epithelial cells, arrest the epithelial cells in G1 phase of cell cycle, dramatically decrease the number of epithelial cells in S and G2 phases; Meanwhile CSE could decrease the expression level of FAK and the degree of its tyrosine phosphorylation. The above effects of CSE were concentration-dependent. The expression of FAK and the degree of its phosphorylation was positively correlated to the increased number of epithelial cells in G1 phase, and negatively to the number of epithelial cells in S and G2 phases. It was concluded that the mechanism by which CSE could inhibit the proliferation of human epithelial cells was contributed to the increased expression and activation of FAK.
Bronchi/*cytology ; Bronchi/metabolism ; Cell Cycle/drug effects ; Cell Proliferation ; Cells, Cultured ; Enzyme Activation ; Epithelial Cells/*cytology ; Epithelial Cells/enzymology ; Focal Adhesion Protein-Tyrosine Kinases/biosynthesis ; Focal Adhesion Protein-Tyrosine Kinases/*metabolism ; Phosphorylation ; Tobacco/adverse effects ; Tobacco Smoke Pollution/*adverse effects

AIM: To study the effect of cigarette smoke extract (CSE) on adhesion and migration of human airway epithelial cells and it's mechanism. METHODS: After 24 h culture, the airway epithelial cells were treated with different concentrations of CSE. The rate of cell attachment and the velocity of cell migration were measured. The expression of FIP200 mRNA and protein were analyzed by RT-PCR and Western blotting. RESULTS: CSE inhibited the rate of cell attachment and the velocity of cell migration. Meanwhile CSE increased the expression level of FIP200 mRNA and FIP200 protein. The effects of CSE became more evident with increased concentration of CSE. Expression of FIP200 mRNA and FIP200 protein were positively correlated to the decreased rate of cell attachment and the velocity of cell migration. CONCLUSION: CSE inhibits the rate of cell attachment, the velocity of cell migration and increases the expression of FIP200.

To examine the deletion and point mutation of WWOX (WW domain containing oxidoreductase) exons 6-8 in human non-small cell lung cancer and their possible relationship with pathological stages, tumor tissues and the corresponding normal tissues were obtained from 44 Chinese patients who had undergone surgery for non-small cell lung cancer. RNA was extracted from each sample and deletion and mutation of WWOX exons 6-8 were analyzed by RT-PCR and DNA sequencing. Our results showed that 28 of 44 (63.6%) lung cancer samples showed loss of WWOX exons 6-8 transcript and the deletion was detected in only 3 of 44 (6.8%) corresponding adjacent normal tissues (P < 0.05). The transcript sequencing analyses of the 16 lung cancer samples without transcript loss of WWOX exons 6-8 revealed no difference from the sequence of GenBank. Moreover, the deletion of WWOX exons 6-8 was significantly higher in the smokers when compared with the non-smokers. It is also higher in the men and squamous carcinomas than in women and adenocarcinomas (P < 0. 05). The deletion, however, was not found to be associated with pathological stages of the tumors. Our study documented a high incidence of deletion of WWOX exons 6-8 in non-small cell lung cancer in Chinese patients and suggested that the frequent loss of WWOX exons 6-8 might play an important role in the tumorigenesis of non-small cell lung cancer in Chinese. WWOX exons 6-8 may serves as a candidate molecular target of smoking carcinogenesis, and point mutation is not a predominant way of alteration of WWOX exons 6-8.
Carcinoma, Non-Small-Cell Lung/*genetics ; Exons/genetics ; Gene Deletion ; Loss of Heterozygosity ; Lung Neoplasms/*genetics ; Oxidoreductases/*genetics ; Point Mutation ; Sequence Analysis, DNA ; Tumor Suppressor Proteins

A mammalian expression plasmid pcDNA3. 1-hCC10 was constructed and identified, then CC10 protein expression in A549 lung cancer cell line was detected. A 273 bp cDNA fragment was amplified from the total RNA of normal lung tissue by using RT-PCR and cloned into expression plasmid cDNA3. 1, and the recombinant plasmid was identified by employing double digestion restriction enzymes Hind III and BamH I and the cDNA sequence was assayed by the Sanger dideoxy-mediated chain termination method. The segment was then transfected into the A549 lung cancer cell line. The protein expression of CC10 was detected by immunofluorescence and Western blot. Our results showed that the cDNA fragment included the entire coding region (273 bp). The recombinant eukaryotic cell expression vector of pcDNA3. 1-hCC10 was successfully constructed, and the sequence of the insert was identical to the published sequence. A549 cells line transfected with the pcDNA3. 1-hCC10 expressed high level of CC10 protein. The recombinant plasmid cDNA3. 1-hCC10 may serve as an effective tool for the study of tumorogenesis and tumor treatment.
Adenocarcinoma/*metabolism ; Adenocarcinoma/pathology ; Cell Line, Tumor ; Genetic Vectors ; Lung Neoplasms/*metabolism ; Lung Neoplasms/pathology ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/genetics ; Transfection ; Uteroglobin/biosynthesis ; Uteroglobin/*genetics

Objective To study whether focal adhesion kinase (FAK) promotes human pulmonary artery smooth cells (HPASMCs) proliferation.Methods Cultured HPASMCs stimulated by fibronectin (40 mg/L) were passively transfected with sense -FAK oligonucleotides(ODNs), FAK activity was measured by immunoprecipitation and expression of FAK protein was detected by Western blots.Meanwhile, the change of cell proliferation was measured by MTT and 3H-TdR absorbation experiment. Results The change of FAK activity and FAK protein content was dose and time dependent at diferential concentration and time passively transfected with sense-FAK ODNs in Cultured HPASMCs. At the same time, sense-FAK ODNs prompoted HPASMCs proliferation and 3H-TdR absorbation.Conclusion FAK can faciliate HPASMCs proliferation,which may play an important function in pulmonary artery hypertension development.

To study the effect of Erigeron breviscapini on chronic hypoxic pulmonary hypertension and mechanism，the hypoxic rats were injected intraperitoneally with Erigeron breviscapini. The mPAP，the content of serum nitric oxide (NO) and endothelin-1 (ET-1) were measured 5，14 and 28 days after intraperitoneal injection of Erigeron breviscapini in hypoxia respectively，and were compared with those in the single hypoxic group. The results showed that the contents of serum mPAP and ET-1 were higher，and serum NO was lower in single hypoxic group than in the normal rats (both P<0.01). The contents of serum mPAP and ET-1 were lower and NO was higher in the Erigeron breviscapini-treated group than in the single hypoxic group (P<0.01). It was suggested that Erigeron breviscapini could reduce the hypoxic pulmonary hypertension by increasing NO and decreasing ET-1 of serum. Erigeron breviscapini could be used to treat hypoxic pulmonary hypertension.

AIM: To investigate the role of potassium channels in the regulation of intracellular free calcium concentration (_i) of pulmonary artery smooth muscle cells (PASMCs) in rats. METHODS: The fluorescence Ca 2+ indicator Fura-2/AM was used to observe _i of rat PASMCs in normal and chronic hypoxic condition. The influences of potassium channels on PASMCs proliferation were assessed by MTT assay. RESULTS: 1. In normoxic condition,_i was (156.91?8.60) nmol/L,and in hypoxic condition,_i was (294.01?16.81) nmol/L. 2. In normoxic condition,the voltage-dependent K +-channel antagonist 4-aminopyridine (4AP),but not the Ca 2+ -activated K +-channel antagonist tetraethylammonium (TEA) and the ATP-sensitive K +-channel antagonist glibenclamide (Glib) increased _i. 3. In hypoxic condition,4AP and TEA caused the rise in _i [(422.41?24.28) nmol/L,(380.84?11.02) nmol/L,respectively],but Glib had no effect on _i. 4. MTT assay showed that 4AP increased the value of absorbing light degree ( A value) in normoxic and hypoxic condition (0.582?0.062,0.873?0.043,respectively, P

AIM: To investigate the effect of sodium nitroprusside (SNP) on activation of nuclear factor ?B. METHODS: The techniques of culture of human T lymphocytes, Western blot and RT-PCR were applied. The effects of NO donor sodium nitroprusside (SNP) at different concentrations on mRNA and protein expression of I?B ? in human T lymphocytes at 30 min or 120 min after stimulating with phytohaemagglutinin (PHA-P) were observed. RESULTS: SNP at middle or high concentrations reduced the degradation of I?B ? protein 30 min after stimulating with PHA-P, and increased the re-expression of I?B ? mRNA 120 min after stimulating with PHA-P significantly. CONCLUSION: The mechanism of inhibitory effect of SNP at middle or high concentrations may be due to the decrease in degradation and the increase in re-synthesis of I?B ?. The regulatory mechanism of SNP at low concentration may not be through I?B ?.