Expression of PPAR-γ in Lung Cancer and the Role of PPAR-γ in Apoptosis of Lung Cancer

Objective: To investigate the relationship between peroxisomeproliferators-activated receptor-gamma (PPAR-γ) and clinical pathology of lung cancer, and to studythe inhibitory effect of PPAR-γ activators on lung cancer growth and the mechanism of inducingapoptosis of lung cancer. Methods: Expression of PPAR-γ in 15 cases of non-cancerous lung tissuesand 64 cases of lung cancer tissues was detected by using immunohistochemistry. The average A valueswere measured by using image analysis. The expression of PPAR-γ in lung cancer cells was detectedby using RT-PCR. After being treated with PPAR-γ activators, apoptosis was detected by using flowcytometry, and meanwhile, the change in caspase-3 activity was detected by caspase-3 kits. Results:Expression levels of PPAR-γ in cancerous tissues were higher than those in non-cancerous lungtissues. In four types of lung cancer, the sequence of PPAR-γ expression from high to low levelswas small-cell lung cancer, squamous carcinom, large-cell lung cancer, adenocarcinoma in turns. Theexpression of PPAR-γ was correlated with differentiation and postoperative TNM staging of lungcancer tissues, but not with lymph node metastasis. PPAR-γ expressed in two lung cancer cell lines,which could induce apoptosis of lung cancer cells after being treated with PPAR-γ activators, andcaspase-3 activity in cells treated with PPAR-γ activators was significantly increased. Conclusion:PPAR-γ is correlated with clinical pathology and apoptosis of lung cancer closely, and activatedPPAR-γ can increase caspase-3 activity to induce cells apoptosis. PPAR-γ will be able to become anew target for therapy of lung cancer in the future.

Construction of Plant-based Expression Vector of Polyphosphate Kinase Gene

[Objective] The aim was to construct the fusion expression vector of polyphosphate kinase（PPK） and green fluorescent protein（GFP） genes.[Method] In this study,the primers were designed based on PPK gene sequence（L03719） of E.coli DH5α in Genbank.Genomic DNA of E.coli DH 5α was extracted as template for the amplification of PPK gene by PCR method.By using In-Fusion@ HD Cloning Kit,the PPK gene was directionally cloned into NcoI site of the pCAMBIA1302 vector.[Result] Sequencing results showed that the 2.0 kb long fragment of PPK gene was inserted into the plant-based expression vector pCAMBIA1302 in front of GFP gene.[Conclusion] The fusion expression vector of PPK and GFP genes were successfully constructed.

Gene Cloning and Expression of PKA Catalytic Subunit and Its Activity in Arabidopsis thaliana

Protein phosphorylation and dephosphorylation are the general means of regulation metabolism within a cell. A PKA catalytic subunit was found in Arabidopsis genome using Blast software. The cDNA was cloned by RT-PCR and sequencing result indicated a high degree of homology at protein level. The cDNA was subcloned into pET30a (+) and expressed in E. coli at different temperatures. The target protein was insoluble when induced at 37degreesC, while dissolvable if induced at 22degreesC with 0.01 mmol/L IPTG. Ni2+-NTA affinity chromatography was used to purify the target protein, which was shown to have cAMP-dependent protein kinase activity. Western blotting analysis indicated that stress treatments affected the expression of PKA catalytic subunit at protein level just to a small extent.

Development of a Transient Assay for Investigating the Activation of Pea Hsp70 Gene Promoter by Potyviral Cistrons

建立了一个瞬时表达系统以研究豌豆种传花叶病毒 (PSbMV ,一种马铃薯Y类病毒 )不同基因对豌豆(PisumsativumL .)Hsp70基因启动子激活能力的差异。构建了豌豆Hsp70基因启动子指导下的GUS基因的表达载体 ,同时还制备了 35S启动子指导下的PSbMVP1和P3基因的表达载体。以表达载体DNA包被的金粉做子弹 ,利用基因枪对豌豆离体叶片进行共轰击实验 ,结果表明 ,PSbMVP1和P3基因在激活豌豆Hsp70基因启动子的能力上存在明显差异。

Overexpression of Aurora-A kinase promotes tumor cell proliferation and inhibits apoptosis in esophageal squamous cell carcinoma cell line

Attrora-A kinase, a serine/threonine protein kinase, is a potential oncogene. Amplification and overexpression of Aurora-A have been found in several types of human tumors, including esophageal squamous cell carcinoma （ESCC）. It has been demonstrated that cells overexpressing Attrora-A are more resistant to cisplatin-induced apoptosis. However, the molecular mechanisms mediating these effects remain largely unknown. In this report, we showed that overexpression of Attrora-A through stable transfection of pEGFP-Aurora-A in human ESCC KYSE150 cells significantly promoted cell proliferation and inhibited cisplatin- or UV irradiation-induced apoptosis. Cleavages of caspase-3 and poly （ADPribose） polymerase （PARP） in Attrora-A overexpressing cells were substantially reduced after cisplatin or UV treatment. Furthermore, we found that silencing of endogenous Aurora-A kinase with siRNA substantially enhanced sensitivity to cisplatin- or UV-induced apoptosis in human ESCC EC9706 cells. In parallel, overexpression of Aurora-A potently upregulated the expression of Bcl-2. Moreover, the knockdown of Bcl-2 by siRNA abrogated the Aurora-A＇s effect on inhibiting apoptosis. Taken together, these data provide evidence that Aurora-A overexpression promoting cell proliferation and inhibiting apoptosis, suggesting a novel mechanism that is closely related to malignant phenotype and anti-cancer drugs resistance of ESCC cells.

Micro-Pulse Lidar Measurements of Aerosol Vertical Structure over the Loess Plateau

Knowledge of the vertical distribution of aerosols in the free troposphere is important for estimating their impact on climate. In this study, direct observations of the vertical distribution of aerosols in the free troposphere are made using surface Micro-Pulse Lidar (MPL) measurements. The MPL measurements were made at the Loess Plateau (35.95°N, 104.1°E), which is near the major dust source regions of the Taklimakan and Gobi deserts. The vertical distribution of the MPL backscattering suggested that nondust aerosols floated from ground level to an altitude of approximately 9 km around the source regions. Early morning hours are characterized by a shallow aerosol layer of a few hundred meters thick. As the day progresses, strong convective eddies transport the aero- sols vertically to more than 1500 m.

Comparison of Visibility Measurements over Horizontal Path by Micro-pulsed Lidar and Visibility Meter

As a powerful tool to scan the atmosphere, the I idar can derive visibility values by directly collecting the backscattering laser light from the atmosphere. Simultaneous measurements of atmospheric visibility by Micro-pulsed lidar （MPL） and a commercial visibility meter （VM） NQ-1 have been performed to evaluate the feasibility of the MPL system designed by the Ocean Remote Sensing Laboratory （ORSL） of the Ocean University of China （OUC） from October 21 2005 to November 21 2005 in the Shilaoren Sightseeing Garden on the Qingdao coast. All the 880 data samples obtained by the two instruments have high correlation coefficients （up to 0.86）, which indicates it is feasible to utilize MPL to measure atmospheric visibility.

Field Detection of Chlorophyll-a Concentration in the Sea Surface Layer by an Airborne Oceanographic Lidar

An airborne oceanographic lidar, with a frequency-tripled Q-switched Nd： YAG laser of 355 nm, has been designed to measure chlorophyll-a （Chl-a） concentration in the sea surface layer by the Ocean Remote Sensing Institute, OUC. The field experiment was carried out in the bay which is located south of the Liaodong Peninsula on the 10th of September 2005. After the flight, the raw data were processed and analyzed by the fluorescence-to-Raman ratio method with seawater attenuation coefficients calculated from signal profiles. The results of Chl-a concentration sea water were also compared with those of Chl-a concentration by measurements by lidar are shown. The measurements in clear a Moderate Resolution Imaging Spectroradiometer （MODIS）.

Observation of nocturnal low-level wind shear and particulate matter in urban Beijing using a Doppler wind lidar

A field experiment of nocturnal mountain wind and corresponding particulate matter （PM） evolution under weak synoptic forcing at five sites within urban Beijing was conducted using a moving Doppler wind lidar and a fixed tower. Clear wind shear and zero-horizontal-wind zones at 40-320 m above the ground with a delay of 1.5 h were found at two sites between 20 km from north to south urban Beijing. The wind speed and height of the low-level jet at the north urban Beijing site were greater than those at the east urban Beijing site. The average horizontal distribution of low-level PM at 240 m was similar to the ground-level PM at night. The PM2.s （aerodynamic diameter ≤2.5 μm） accumulation center showed no abrupt changes with a shift in wind direction until the northerly wind jet arrived.

Correlation between expression of gastrin, somatostatin and cell apoptosis regulation gene bcl-2/bax in large intestine carcinoma

AIM: To explore the correlation between expression of somatostatin (SS), gastrin (GAS) and cell apoptosis regulation gene bcl-2/bax in large intestine carcinoma.METHODS: Sixty-two large intestine cancer tissue samples were randomly and retrospectively selected from patients with large intestine carcinoma. Immunohistochemical staining for bcl-2, bax, GAS, SS was performed according to the standard streptavidin-biotin-peroxidase (S-P) method.According to the semi-quantitative integral evaluation, SS and GAS were divided into three groups as follows. Scores1-3 were defined as the low expression group, 4-8 as the intermediate expression group, 9-16 as the high expression group. Bax and bcl-2 protein expressions in different GAS and SS expression groups of large intestine carcinoma were assessed.RESULTS: The positive expression rate of bax had a prominent difference between SS and GAS high, intermediate and low expression groups (P＜0.05, x2ss = 9.246; P＜0.05,x2GAS = 6.981). The positive expression rate of bax in SS high (80.0%, 8/10) and intermediate (76.5%, 13/17)expression groups was higher than that in low expression group (40.0%, 14/35) (P＜0.05, x2high vs low = 5.242; P＜0.05,x2middle vs low = 6.097). The positive expression rate of bax in GAS high expression group (27.3%, 3/8) was lower than that in low expression group (69.4%, 25/36) (P＜0.05,x2 = 4.594). However, bax expression in GAS intermediate expression group (46.7%, 7/15) was lower than that in low expression group, but not statistically significant. The positive expression rate of bcl-2 had a prominent difference between SS and GAS high, intermediate and low expression groups (P＜0.05, x2ss = 7.178; P＜0.05, x2GAS = 13.831). The positive expression rate of bcl-2 in GAS high (90.9%, 10/11)and intermediate (86.7%, 13/15) expression groups was higher than that in low expression group (44.4%, 16/36)(P＜0.05,x2high vs low = 5.600; P＜0.05, x2 middle vs low = 7.695).However, the positive expression rate of bcl-2 in SS high (40.0%, 4/10) and intermediate (47.1%, 8/9) expression groups was lower than that in low expression group (77.1%, 27/35)(P＜0.05, x2 high vs low = 4.710; P＜0.05, x2 middle vs low = 4.706).There was a significant positive correlation between the integral ratio of GAS to SS and the integral of bcl-2 (P＜0.01,r=0.340). However, there was a negative correlation between the integral ratio of GAS to the SS and bax the integral of (P＜0.05, r = -0.299).CONCLUSION: The regulation and control of gastrin,somatostatin in cell apoptosis of large intestine carcinoma may be directly related to the abnormal expression of bcl-2, bax.

Polo-like kinase 1 expression is a prognostic factor in human colon cancer

AIM：To clarify the expression patterns and prognostic implications of the mitotic regulator Polo-like kinase 1 （PLK1） in colon cancer. METHODS： Expression of PLK1 was investigated by immunohistochemistry （158 cases） and immunoblotting in tissue of colon adenomas and adenocarcinomas. PLK1 expression patterns were correlated with clinicopathological parameters and patient prognosis. In addition, expression of PLK1 was evaluated by immunoblot and PCR in colon carcinoma cell lines, and coexpression of PLK1 with the proliferation marker Ki-67 was investigated. RESULTS： Weak PLK1 expression was observed in normal colon mucosa and adenomas. In contrast, 66.7% of carcinomas showed strong expression of PLK1. Overexpression of PLK1 correlated positively with Dukes stage （P〈0.001）, tumor stage （P = 0.001） and nodal status （P〈0.05）. Additionally, PLK1 expression was a prognostic marker in univariate survival analysis （P〈0.01） and had independent prognostic significance （RR = 3.3, P = 0.02） in patients with Iocoregional disease. Expression of PLK1 mRNA and protein was detected in all cell lines investigated. Coexpression of PLK1 and Ki-67 was observed in the majority of colon cancer cells, but a considerable proportion of cells showed PLK1 positivity without Ki-67 expression. CONCLUSION： PLK1 is a new prognostic marker for colon carcinoma patients and may be involved in tumorigenesis and progression of colon cancer. Strategies focusing on PLK1 inhibitionin vivo might therefore represent a promising new therapeutic approach for this tumor entity. 2005 The WIG Press and Elsevier Inc. All rights reserved

Laser diode drive method with narrow-width and high-peak current for multi-line LIDAR

Light detection and ranging (LIDAR) based on time of flight (TOF) method is widely used in many fields related to distance measurement. LIDAR generally uses laser diode (LD) to emit the pulsed laser with high peak power and short duration to ensure a large distance measurement range and eye safety. To achieve this goal, we propose a pulsed LD drive method producing the drive current with high peak and narrow pulse width. We analyze the key issues and related theories of the drive current generation based on this method and design an LD driver. A model of drive current generation is established and the influence of operating frequency on drive current is discussed. The LD driver is simulated by software and verified by experiments. The working frequency of the driver changes from 20 kHz to 100 kHz and the charging voltage is set at 130 V. The current produced by this driver has a duration of 8.8 ns and a peak of about 35 A, and the peak output optical power of the LD exceeds 75 W.

Accuracy assessment of mine walls＇ surface models derived from terrestrial laser scanning

The monitoring of highwall slopes at open-pit mines is an important task to ensure safe mining. For this reason, several techniques such as total station, radar, terrestrial Light Detection and Ranging （LIDAR） can be employed for surface measurement. The objective of this study is to investigate mesh algorithms, which can be used to interpolate 3D models of pit walls. Experiments were carried out at Coc Sau open-pit mine at Quang Ninh province of Vietnam, and at experimental mine of Akademia Gtrniczo-Hutnicza University of Science and Technology in Cracow, Poland. First, 3D point cloud data for the study area was acquired by using terrestrial LIDAR, then was used to generate mesh surfaces using three algorithms--Delaunay 2.5D XY Plane, Delaunay 2.5D Best Fitting Plane, and Mesh from Points. After that, the results were rectified and optimized. Subsequently, the optimized meshes were used for generation of non-uniform rational basis spline （NURBS） surfaces. Then, the NURBS surface accuracy was assessed. The results showed that the average distance between surface and point cloud was within range of 5.6-5.8 mm with deviation of 6.2-6.8 mm, depending on the used mesh. Additionally, the quality of surfaces depends on the quality of input data set and the algorithm used to generate mesh network, and the accuracy of computed NURBS surfaces fitting into pointset was 4-5 times lower than that of optimized mesh fitting. However, the accuracy of the final product allows determining displacements on the level of centimeters.

Estimating underground mine ventilation friction factors from low density 3D data acquired by a moving LiDAR

Ventilation system analysis for underground mines has remained mostly unchanged since the Atkinson method was made popular by Mc Elroy in 1935. Data available to ventilation technicians and engineers is typically limited to the quantity of air moving through any given heading. Because computer-aided modelling, simulation, and ventilation system design tools have improved, it is now important to ensure that developed models have the most accurate information possible. This paper presents a new technique for estimating underground drift friction factors that works by processing 3 D point cloud data obtained by using a mobile Li DAR. Presented are field results that compare the proposed approach with previously published algorithms, as well as with manually acquired measurements.

The effect of C-terminal fragment of JNK2 on the stability of p53 and cell proliferation

The basal activity of JNK is low in normal growing cells and inactivated JNK targets p53 for ubiquitination. To elucidate if the C-terminal part of JNK is responsible for its binding to p53, the low background tet-off inducible NIH3T3 cell line was selected by luciferase reporter gene and a double stable C-JNK Aa (203-424) cell line was established. After withdrawing tetracycline, the C-JNK fragment expression was induced and cell growth was dramati- cally inhibited 24 h later. However, the expresion of p53 was found to be increased after the induction of C-JNK fragment, evaluated by transfecting p21waf-luciferase reporter genes. Our further studies showed that C-JNK fragment could form complex with p53 both in vivo and in vitro. Induction of C-JNK fragment in vivo can increase p53 stability by inhibiting p53 ubiquitination.

Gene Expression Controlled by Heat-Inducible Site-Specific Recombination in Tobacco

Cre site-specific recombinase-mediated DNA excision system was driven by the heat shock promoter Gmhsp17.5C. In this system, the DNA fragment with CaMV35S-GUS franked by two identical orientation loxp sites could be excised from the transgenic tobacco (Nicotiana tabacum L. cv. W38) by Cre expression under control of heat shock promoter. This transgenic system has been determined by quantitative PCR and showed Cre/lox mediated recombination efficiency. Results showed that 41% of DNA fragment with CaMV35S-GUS in the transgenic tobacco could be excised after a two-hour heat shock treatment. Based on several advantages of heat shock-inducible site-specific recombination system such as easy manipulation, sensitivity to heat shock and no background expression, it can be potentially used for inducible DNA manipulation in transgenic plant.

Correlation between the expressions of gastrin, somatostatin and cyclin and cyclin-depend kinase in colorectal cancer

AIM： To explore the correlation between the expressions of gastrin （GAS）, somatostatin （SS） and cyclin, cyclin- dependent kinase （CDK） in colorectal cancer, and to detect the specific regulatory sites where gastrointestinal hormone regulates cell proliferati6n. METHODS： Seventy-nine resected large intestine carcinomatous specimens were randomly selected. Immunohistochemical staining for GAS, SS, cyclin D1, cyclin E, cyclin A, cyclin B1, CDK2 and CDK4 was performed according to the standard streptavidinbiotin-peroxidase （S-P） method. According to the semiquantitative integral evaluation, SS and GAS were divided into high, middle and low groups. Cyclin D1, cyclin E, cyclin A, cydin B1, CDK2, CDK4 expressions in the three GAS and SS groups were assessed. RESULTS： The positive expression rate of cyclin D1 was significantly higher in high （78.6%, 11/14） and middle GAS groups （73.9%, 17/23） than in low GAS group （45.2%, 19/42） （P〈0.05, X^2 high vs low = 4.691; P〈0.05, X^2 middle vs low = 4.945）. The positive expression rate of cyclin A was significantly higher in high （100%, 14/14） and middle GAS groups （82.6%, 19/23） than in low GAS group （54.8%, 23/42） （P〈0.01, X2high vs low = 9.586; P〈0.05, X^2 middle vs low = 5.040）. The positive expression rate of CDK2 was significantly higher in high （92.9%, 13/14） and middle GAS groups （87.0%, 20/23） than in low GAS group （50.0%, 21142） （P〈0.01, X^2 high vs low = 8.086; P〈0.01,X^2 middle low = 8.715）. The positive expression rate of CDK4 was significantly higher in high （78.6%, 11/14）and middle GAS groups （78.3%, 18/23） than in low GAS group （42.9%, 18/42） （P〈0.05, X^2 high vs low= 5.364; P〈0.01, X^2 middle vs low = 7.539）. The positive expression rate of cyclin E was prominently higher in low SS group （53.3%, 24/45） than in high （9.1%, 1/11） and middle （21.7%, 5/23） SS groups （P〈0.05, X^2 high vs low = 5.325; P〈0.05, X^2 middle vs low = 6.212）. The positive expression rate of CDK2 was significantly higher in low SS group （77.8%, 35/45） than in high SS group （27.3%, 3/11） （P〈0.01, X^2 high vs low = 8.151）. There was a significant positive correlation between the integral ratio of GAS to SS and the semi-quantitative integral of cyclin D1, cyclin E, cyclin A, CDK2, CDK4 （P〈0.05, 0% = 0.252; P〈0.01, E^rs = 0.387; P〈0.01,A^rs = 0.466; P〈0.01, K2^rs = 0.519; P〈0.01, K4^rs = 0.434）. CONCLUSION： The regulation and control of gastrin, SS in colorectal cancer cell growth may be directly related to the abnormal expressions of cyclins D1, A, E, and CDK2, CDK4. The regulatory site of GAS in the cell cycle of colorectal carcinoma may be at the G2, S and G2 phases. The regulatory site of SS may be at the entrance of S phase.

SELECTION OF BRILLOUIN SHIFT DISCRIMINATOR FOR BRILLOUIN LIDAR

For the measurement of vertical profiles of sound speed in the sea using laser excited Brillouin scattering, a high resolution measurement of Brillouin frequency shift is required. In this work, a molecular absorption cell was selected as the frequency shift discriminator and several kinds of absorption gases were tried. It was found that the strong line (#1095) of 127 I 2 at 18783.3297 cm -1 and two absorption lines of 129 I 2 located at the two sides of the #1095 line of 127 I 2 could be used as frequency shift discriminator to detect the changes of the Brillouin frequency shift. This selection is the best one within the range from 532.0131 nm to 532.5154 nm. But it is not perfect and there is a lot of work to do before its practical application.

Initial Results of Lidar Measured Middle Atmosphere Temperatures over Tibetan Plateau

During August 2013,a mobile Rayleigh lidar was deployed in Lhasa,Tibet(29.6°N,91.0°E) for making measurements of middle atmosphere densities and temperatures from 30 to 90 km.In this paper,the authors present the initial results from this scientific campaign,Middle Atmosphere Remote Mobile Observatory in Tibet(MARMOT),and compared the results to the MSIS-00(Mass Spectrometer and Incoherent Scatter) model.This work will advance our understanding of middle atmosphere dynamic processes,especially over the Tibetan Plateau area.

Gene expression profiling of gastric cancer by microarray combined with laser capture microdissection

AIM： To examine the gene expression profile of gastric cancer （GC） by combination of laser capture microdissection （LCM） and microarray and to correlate the profiling with histological subtypes. METHODS： Using LCM, pure cancer cells were procured from 45 cancerous tissues. After procurement of about 5 000 cells, total RNA was extracted and the quality of RNA was determined before further amplification and hybridization. One microgram of amplified RNA was converted to cDNA and hybridized to cDNA microarray. RESULTS： Among 45 cases, only 21 were qualified for their RNAs. A total of 62 arrays were performed. These included 42 arrays for cancer （21 cases with dyeswab duplication） and 20 arrays for non-tumorous cells （10 cases with dye-swab duplication） with universal reference. Analyzed data showed 504 genes were differentially expressed and could distinguish cancerous and non-cancerous groups with more than 99% accuracy. Of the 504 genes, trefoil factors 1, 2, and 3 were in the list and their expression patterns were consistent with previous reports. Immunohistochemical staining of trefoil factor 1 was also consistent with the array data. Analyses of the tumor group with these 504 genes showed that there were 3 subgroups of GC that did not correspond to any current classification system, including Lauren＇s dassification. CONCLUSION： By using LCM, linear amplification of RNA, and cDNA microarray, we have identified a panel of genes that have the power to discriminate between GC and non-cancer groups. The new molecular classification and the identified novel genes in gastric carcinogenesis deserve further investigations to elucidate their clinicopathological significance.