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 ?.

AIM: To study the role of hypoxia-inducible factor-1alpha(HIF-1?) on lung cancer cells A549 growth in vitro and in vivo. METHODS: To observe the growth rate of A549 cells after HIF-1? transfected, A549 cells (1?10~6/mouse) were inoculated subcutaneously into 20 nude mice, which were randomly divided into two groups: the control group (group A, n=10), the HIF-1? transfected group (group B, n=10). The weights of subcutaneous tumor were detected. The resected specimens were made into paraffin-embedded sections. The proliferating cell nuclear antigen (PCNA) was identified by immunohistochemistry(ISH). The expressions of HIF-1?? apoptosis-related protein survivin and bcl-2 were analyzed by Western blot. RESULTS: The growth rates of the HIF-1? transfected lung cancer cells A549 were significantly increased, and more importantly, the HIF-1? transfected lung cancer cells A549 was able to enhance lung cancer growth in nude mice(P

AIM: To investigate the effects of NF-?B decoy oligodeoxynucleotides (ODNs) on apoptosis in lung cancer cell A549. METHODS: The treatments of lung cancer cells (A549) were divided into three groups: group A (control group); group B (decoy ODN group) and group C (scramble decoy ODN group). FITC-labeled NF-?B decoy ODNs was transfected into A549 with LipofectAMINE TM 2000. The activation was observed by electrophoretic mobility shift assays (EMSA). The proliferation was observed by growth curve. The apoptosis of cells were observed by flow cytometry and TdT mediated dUTP-biotin Nick End Labeling (TUNEL). The expression of Bcl-2 and Fas were observed by Western blot. RESULTS: After FITC-labeled decoy ODNs was transfected for 1 hour, the decoy ODNs was detected in the nuclei of A549 cells. EMSA performed the depression of the NF-?B binding to the nucleus. The growth curve showed the inhibition of the A549 cell growth and the percentage of apoptosis was increased compare with control group by flow cytometry and TUNEL. The amount of apoptosis inhibitor (Bcl-2) in group A and group C were 2.0 times and 2.1 times more than that in group B, respectively. The level of apoptosis accelerator (Fas) in group B were 2.6 times and 2.3 times more than that in group A and group C, respectively via Western blot. CONCLUSION: The NF-?B decoy ODNs accelerate the apoptosis of lung cancer cell A549 and the mechanism may be due to its inhibiting the expression of Bcl-2 and increasing the level of Fas. [

The morphological variations and congenital malformations of the auditoryossicles are not so rare as previously considered.The congenital anomalies of themhave sometimes been documented in the periodicals of the otorhinolaryngology,buttheir variations reported are scanty and incomplete.With dissecting microscope,weobserved 588 ossicles(200 mallei,224 incudis 164 stapes)from 120 full term foe-tuses without congenital defect except one case of anencephaly.We found that theauditory ossicles vary greatly in their form,length,size,angulation,curvature orthickness,etc.Those are variations of anterior fovea of caput mallei,anteriorcurvature of manubrium mallei,form of lateral margin of manubrium mallei,formof crus breve of incus,form of crus stapedis and patterns of basis stapedis.In 240 ears,five cases of congenital malformations of ossicular chain(2.1%)were discovered and listed as follows:1.One case of congenital stapes footplate fixation,2.One case of ring form stapes detached from the basis stapedis,3.Two cases of columella stapes,4.One case of triple fusion of ossicles by osteoid tissue.Embryology of auditory ossicles available for understanding the variability anddeformity was briefly reviewed.According to our investigations we come to the conclusion that the stapesis probably the most frequently involved in the morphological variation andmalformation.

The arteries of auditory ossicles of 40 ears from 20 full term feotuses weredemonstrated by injection of liquid latex containing a small amount of Chinese inkthrough common carotid arteries.We found that the malleus and incus possess thenutrient arteries as well as the mucosal arteries,whereas the stapes gets its bloodsupply from the mucosal arteries only.The anterior tympanic artery is the principal source of blood supply of the mal-leus and the incus.It enters the middle ear through the petrotympanic fissure andramifies into five branches:malleolar artery,incudal artery,superior branch,posteriorbranch and chorda tympani branch.The malleolar and incudal arteries are nutrientarteries.The vascular network in mucosa over the manubrium mallei is supplied bythe branches of the deep auricular and stylomastoid arteries over the tympanic mem-brane.The mucosal arteries of the long crus of the incus is supplied by the smallvessels given off by incudal artery before entering the nutrient foramen,the finevessels from the arteries around the chorda tympani and the vessels passing to it fromstapes.The blood supply of the stapes is derived from the vessels located in two majorareas:one from the facial canal and the other from the promontory.The arteries tothe stapes from the promontory vascular plexus are the artery of the head of thestapes,the artery of the posterior crus and the artery of the anterior crus.The for-mer two vessels have not been reported previously.In the facial canal there are thestylomastoid artery and the superficial petrosal artery.The arterial supply of incudostapedial joint and the distal portion of the incuscomes from the vessels passing to them from the stapes rather than from the incudalsource.From above account,it would appear that the head of the malleus and thebody and short crus of the incus derived from first branchial cartilage are mainlysupplied by the anterior tympanic artery,and the remainder of the auditory ossiclesderived from second branchial cartilage are supplied by the stylomastoid artery.

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

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

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.