In order to construct plasmid of hypoxia-inducible factor-1alpha (HIF-1alpha), and transfect into human lung cancer cells A549, the change in sensitivity of lung cancer cells A549 to chemotherapy was observed. HIF-1alpha mRNA structure region was amplified by RT-PCR and inserted into plasmid pcDNA3. The expression plasmid pcDNA3/HIF-1alpha was transfected into A549 with Lipofec-tAMINE2000. The expression of HIF-1alpha protein was detected by Western blot. After A549 cells were transfected with HIF-1alpha prior to addition of 5-Fu, the growth activity was measured by growth curve, apoptosis was detected by flow cytometry at 48 h, and the levels of caspase3 and MDR-1 were determined by Western blot. The results showed that the constructed expression plasmid was analyzed with restriction enzymes and gel electrophoresis. Two DNA lanes at 2.55 kb and 5.4 kb respectively were found, which were consistent with that expected. The growth rate in 5-Fu group was significantly inhibited, and the apoptosis index and caspase3 activity were increased significantly as compared with control group. After HIF-1alpha being transfected into A549, the activity of MDR-1 was increased and the effect of 5-Fu was weakened. In conclusion, HIF-1alpha can promote chemoresistance by increasing the activation of MDRI and suppressing apoptosis during lung cancer cells A549 induced with 5-Fu.

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. [

To study the role and mechanisms of hypoxia-inducible factor-1alpha (HIF-1alpha) on the growth and tumorigenicity of lung cancer cells A549, the antisense oligonucleotide of HIF-1alpha was transfected to A549 cells. The effect of the antisense oligonucleotide on tumor growth in vitro and in vivo was evaluated by the growth rate suppression of A549 cells and subcutaneous implanted tumor in nude mice, and the effect on tumorigenicity was evaluated by the expression inhibition of angiogenic factors, the microvessel density (MVD) and vascular endothelial growth factor (VEGF) protein expression which were detected by immohistochemistry and western blot respectively. This study revealed that in vitro the growth rate of antisense oligonucleotide group was significantly decreased as compared with that of control group, sense oligonucleotide group and false-sense oligonucleotide group; in vivo the weight of implanted tumors in nude mice of antisense oligonucleotide group was 1.51 +/- 0.40 g, which was significantly lower than that of control group (2.79 +/- 0.33 g), sense oligonucleotide group (2.81 +/- 0.45 g) and false-sense oligonucleotide group (2.89 +/- 0.39 g) and the inhibitory rate was 47%. Both MVD and VEGF protein expression were significantly inhibited in antisense oligonucleotide group compared with those in other groups. These results indicated that antisense oligonucleotide of HIF-1alpha could inhibit lung cancer cells A549 growth in vitro and in vivo, and the mechanism may be due to the inhibition of vascular growth and VEGF protein expression.

Objective To compare the efficacy between microwave endometrial ablation(MEA) and abdominal hysterectomy(AH) in the treatment of abnormal uterine bleeding(AUB).Methods A total of 80 patients with AUB was divided into two groups,either receiving MEA(40 patients) or receiving AH(40 patients).The clinical efficacy,operation time,intraoperative blood loss,white blood cell(WBC) count,and neutrophil granulocyte rate(NGR) before and after operation were compared.Results The effective rate for abnormal uterine bleeding was not significantly different between MEA(97.5%,39/40) and AH(100%,40/40)(?2=0.000,P=1.000).As compared with AH,MEA was characterized by a shorter operation time(4.8?1.2 min vs 102.5?34.6 min;t=17.848,P=0.000) and less intraoperative blood loss(0 ml vs 138.3?40.6 ml;t=21.544,P=0.000).Conclusions Microwave endometrial ablation is a novel and efficient minimally invasive method for abnormal uterine bleeding.

Objective To evaluate the treatment results and side-reactions of esophageal carcinoma with late course hyperfractionated radiotherapy ( LCHR) plus different chemotherapy. Methods A prospective research was carried out on 287 advanced stage esophageal carcinoma patients whom were randomized into there groups;LCHR + cisplatin +5-fluorouracil +leucovorin group( A group), LCHR +5-fluorouracil polyphase lipo- some group (B group) and LCHR group(C group). 115 patients were in A group and 107 patients in B group and 65 patients in C group. Radiotherapy method; there groups were treated by conventional fractionated radio- therapy during the first two-thirds of the whole course with 40 Cy,then followed by isocenter hyperfractionated radiotherapy to keep away of spinal marrow, 1. 3-1. 5Gy per time,2 times a day and 5 days a week. The total dose was 60-66 Gy in A, B group and 60-70 Gy in C group. The preventive dose was 50 Gy. Results There was no statistically difference among the there groups on the short-term curative effect The 1- ,2- ,3-year local control rate was 80% ,50.4% ,42.6% and 72.9% ,51.4% ,41. 1% and 63.1% ,38.5% ,30.8%

To establish a better method of primary culture for alveolar epithelial type II cells (AEC II) and to study its bionomics, alveolar epithelial type II cells were isolated by digestion with trypsin and collagenase, which were then purified by plated into culture flask coated with rat immunoglobulin G. The purified AEC II were identified by alkaline phosphatase staining, electron microscopy, immunocytochemical staining of pulmonary surfactant protein A (SPA). The SPA expression and transfection characteristics were compared with those of A549 cell line. The results showed that AEC II could be isolated by digestion with trysin and collagenase and purified by adhesive purification by using IgG, with a yield of about 2-3 x 10(7), and a purity of about 75%-84%. Cells could be quickly identified with AKP staining. AEC II were different from A549 cell line in terms of SPA expression and transfection characteristics. It is concluded that adhesive purification with IgG can improve the purity of AEC II, and AKP staining is simple in cell identification. AEC II can not be completely replaced by A549 cells in some studies because the differences between them, such as SPA expression.
Cell Culture Techniques/*methods ; Cell Separation/*methods ; Cells, Cultured ; Ecology ; Epithelial Cells/*cytology ; Immunoglobulin G/pharmacology ; Pulmonary Alveoli/*cytology ; Pulmonary Surfactant-Associated Protein A/biosynthesis ; Rats, Wistar

To explore the teaching model and effect of emergency care simulator(ECS) teaching in "Shock"during clinical bed side teaching,fifty-six students were randomly divided into two groups, receiving model and traditional teaching model respectively. After one semester,the students’examination results and general effect were evaluated. The students’ex-amination results,clinical theory and clinical skills in ECS teaching group were significantly better than traditional teaching group.

Objective To explore the influence of Hedysari radix concentrations fluid (HRCF) on learning and memory and its effect on cerebral monoamine neurotransmitters in senescence accelerated mice. Methods The senescence accelerated mice were randomly divided into the senescence accelerated mouse/resistance 1 (SAMR1) control group, senescence accelerated mouse/prone-8 (SAMP8) model group, HRCF treatment group and aricept treatment group. After 3 months of intragastrical treatment, the learning and memory ability changes and content of norepinephrine, dopamine, 5-hydroxytryptamine and 5-hydroxyindoleacetic acid were detected respectively using Morris water maze and high performance liquid chromatography. Results In the model group, the hidden platform test indicated that the latency to find the hidden platform was remarkably prolonged, and the content of norepinephrine, dopamine, 5-hydroxytryptamine as well as 5-hydroxyindoleacetic acid were significantly reduced (P<0.01). Compared with the model group, the latency of hidden platform test was remarkably shortened (P<0.05) and the content of norepinephrine, dopamine, 5-hydroxytryptamine as well as 5-hydroxyindoleacetic acid were obviously increased in HRCF treatment group (P<0.01). Conclusion HRCF could improve learning and memory ability of SAMP8 and the influence may related to increasing the content of cerebral monoamine neurotransmitters.

Objective To explore the angiographic characteristic of lower extremities of patients with diabetic foot. Methods The angiographic results of 35 lower extremities of diabetic patients, the characteristics of respective clinical stages and the stenosis degree at different arterial segments were undergone statistical analysis. Results (1) A lotal of 210 arterial segments were divided into the above-knee and the below-knee section. There was significant difference in stenosis degree between the two sections (P