Objective To investigate the expression and regulation of ASPP family mRNA in NSCLC cell lines.and study the diagnostic and therapeutic significances of apoptosis stimulating of ps3 expression in NSCLC Patients.Methods Real-time fluorescence quantitative PCR wag established and used to measure the expression Ievels of ASPP mRNA in NSCLC cell lines A549(wild-type p53)and NCI-H157 (mutant-type p53)with p53 different genetic background,and ASPP mRNA was also detected in NSCLC tissue,peripheral blood of 37 NSCLC patients.Western Bloting was used to examine the levels of ASPP1 and ASPP2 proteins of NSCLC cell Iines.Chemosensitivity of the cell lines to CDDP was analyzed by MTT assay,and detect the change of ASPP1 and ASPP2.Compared with the ASPPs mRNA expression between NSCLC patients and healthy controls.and monitoring ASPPs mRNA expression in patients with NSCLC chemotherapy.Results The slope rotes of standard curves were-3.249,-3.358 respectively.and correlation coefficients were more than 0.98.Amplification efficiency of standard curves were 1.156.1.028 respectively.The IC_(50) values of NCI-H157 and A549 cells were 3.70.10.48 μg/ml respectively.Mean ASPP1 ASPP2 mRNA levels were a statistically significant 2.66 folds and 6.98 folds higher in NCI-H157 cell compared to A549 cell The levels of ASPP1 mRNA in tumor tissues and peripheral blood were (4.27±0.57)×10~3,(2.49±0.32)×10~3 in patients with NSCLC respectively,and the levels of ASPP2 mRNA in tumor tissues and peripheral blood were(2.34±0.75)×10~3,(7.00±1.17)×10~3 in Patients with NSCLC respectively.The levels of ASPP1 and ASPP2 mRNA in tissues and peripheral blood were(1.32±0.21)×10~4,(1.46±0.31)×10~4 and(1.38±0.19)×10~4,(1.28±0.18)×10~4 in control groups respectively.The ASPP1,ASPP2 mRNA levels in patients with NSCLC were significantly lower than those of control groups(t=2.58,3.94,3.62,3.76,P＜0.05).Before chemotherapy the levels of ASPP1 and ASPP2 mRNA in patients with NSCLc were(2.34±0.56)×10~3 and(6.64±0.72)×10~3,and after 2 periods of postoperative chemotherapy the levels were(3.66±0.64)×10~3 and(9.42±0.44)×10~3,the expression was higher than those of ehemotherapy before(t=3.02,4.50,P＜0.05).Conclusions The expression of ASPP1.ASPP2 mRNA in p53 mutant-type cell lines is higher than that in p53 wild-type cell lines.The expression of ASPP maybe has close relationship with chemotherapeutic sensitivity of NSCLC.In NSCLC patient chemotherapy process,the change of ASPP1 and ASPP2 mRNA expression may be related with chemotherapy medicine function.Enhance the expression of ASPP1 and ASPP2 will be beneficial to the tumot treatment.

Objective To study the expression of interleukin-1β in aortic dissections and aneurysms. Methods Aortic specimens were obtained from patients with type Ⅰ thoracic aortic dissection (11 cases),ascending thoracic aortic aneurysms (10 cases),and healthy organ donors (7 cases).Expression of interleukin-1β,matrix metalloproteinase-9,and signal transduction factors phospho-p38 and phospho-JNK were detected by real time RT-PCR,Western blot,and immunohistochemistry,respectively.TUNEL staining was performed to detect apoptosis of media cells. Results Apoptosis in the media of thoracic aortic dissection and ascending thoracic aortic aneurysms was dramatically higher than control group.Expression of interleukin-1β gradually increased in an order of control group,thoracic aortic dissection to ascending thoracic aortic aneurysms ( P ＜ 0.01,respectively).Expression of matrix metalloproteinase-9significantly increased in the media of thoracic aortic dissection and ascending thoracic aortic aneurysms compared with control group (P ＜ 0.01,respectively).There were positive correlations between interleukin1 β and matrix metalloproteinase-9,interleukin-1β and phospho-p38 in thoracic aortic dissection ( P ＜ 0.01,respectively),interleukin-1β and apoptosis in ascending thoracic aortic aneurysms (P ＜ 0.01 ).Conclusions Interleukin-1β and interferon-γ might effect the formation of thoracic aortic dissection and ascending thoracic aortic aneurysms possibly through the up-regulation of matrix metalloproteinase-9 and apoptosis of media cells in humans.

Objective To explore whether the low-intensity pulsed ultrasound (LIPU) could induce apoptosis on SMMC-7721 cells and to explore the underlying mechanism.Methods The SMMC-7721 cells were randomly divided into 4 groups:a blank control group,which was subject to sham exposure to ultrasound,and 3 ultrasound intervention groups exposed to ultrasound at intensities of 0.5,1.3 and 2.0 W/cm2,respectively.Then they were incubated for 6 h.The cell apoptosis,necrosis and changes of cell cycles were measured using the flow cytometry.The transmission electron microscope (TEM) was used to observe microstructural changes in the cells.The agarose gel electrophoresis (AGE) was used to examine the DNA fragmentation,and Western-blotting was employed to assess the protein expression of caspase-3.Results The average cell apoptosis rate of the 3 intervention groups were 4.66％,8.99％ and 32.41％,respectively.The percentage of cells in G2 phase increased significantly and those in G1 phase decreased significantly in the 3 intervention groups compared to the blank control group at the same time points.In the intervention groups,significant cell apoptosis was observed under TEM,and DNA ladders was seen in AGE,with DNA fragments appearing obviously when cells were incubated for 6 h and 9 h after ultrasound exposure.In intervention groups subject to 1.3 and 2.0 W/cm2 ultrasound exposure,the protein expression of caspase-3 was significantly higher than that of the control group.Conclusion LIPU can inhibit the proliferation and induce apoptosis of SMMC-7721 cells with a dose-dependent feature.The possible mechanism underlying the LIPU-induced cell apoptosis might be related to the activation of the mitochondria pathway,and especially the caspase-3 protein.

Background and purpose:Low intensity ultrasound (LIUS) can kill cancer cells and promote their apoptosis. However, it is still unknown how it affects the migration and invasion of tumor cells. This study aimed to explore the effect of LIUS on human hepatocellular line MHCC97H in migration and metastasis and the possible mechanismin vitro.Methods:According to the intensity of ultrasonic irradiation, 4 experimental groups were established: control group (0 W/cm2), 0.5 W/cm2, 1.0 W/cm2 and 1.5 W/cm2group. The migration and invasion ability of hepatocellular cells was detected by scratch assay and Transwell migration and invasion assay after the irradiation of LIUS. The changes of cytoskeleton after irradiation were observed by microscope and F-actin green lfuorescence staining. The expressions of MMP-2 and MMP-9 were examined by real-time lfuorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot.Results:Low intensity ultrasound (≤1.5 W/cm2) promoted the migration and invasion of hepatocellular line MHCC97H. Scratch assay and Transwell assay showed much more cells under irradiation migrated through membrane than untreated. It was found that morphology of liver cancer cells changed after LIUS irradiation using optical microscope and lfuorescence microscope. The results of RTFQ-PCR and Western blot showed upregulation of MMP-2 expression by LIUS in MHCC97H and high expression of MMP-9 mRNA. Conclusion:Low intensity ultrasound may promote the migration and invasion of MHCC97H through changing cytoskeleton and upregulating protein expression of MMP-2.

Objective:To explore any effect of combining intermittent theta-burst stimulation (iTBS) of the cerebellum with physiotherapy on the balance function and gait of stroke survivors.Methods:Thirty-two hemiplegic stroke survivors were divided at random into a treatment group and a control group, each of 16. Both groups received conventional physical therapy. Before their physiotherapy sessions the treatment group received iTBS treatment of the cerebellar hemisphere contralateral to the affected cerebral hemisphere, while the control group was given pseudo-stimulation on the same site. The iTBS was given once a day for 200s each time, 6 times a week for 3 weeks consecutively. Before and after the treatment, as well as 3 weeks later, both groups′ balance was evaluated using the Berg Balance Scale (BBS). Their ability to shift their center of gravity, total length of their shaking trajectory, and maximum shaking diameter were also quantified. Walking ability was assessed using 10m walk test (10MWT) times and the Tinetti Gait Assessment Scale (POMA-G). Lower limb motor function was quantified using the relevant Fugl-Meyer assessment (FMA-LE) and the subjects′ ability in the activities of daily living was measured with the Barthel index (BI).Results:After the 3 weeks of treatment and at the follow-up the average BBS score of the treatment group had improved significantly more than the control group′s average, as had its total track length and maximum shake diameter. The average POMA-G, FMA-LE and BI scores of the treatment group were also significantly better.Conclusions:Combining iTBS with physiotherapy can improve the balance and gait of stroke survivors more effectively than physiotherapy alone.