Down-regulation of Hsp90 could change cell cycle distribution and increase drug sensitivity of tumor cells

AIM:To construct Hsp90 antisense RNA eukaryotic expression vector, transfect it into SGC7901 and SGC7901/VCR of MDR-type human gastric cancer cell lines, HCC7402 of human hepatic cancer and Ec109 of human esophageal cancer cell lines, and to study the cell cycle distribution of the gene transected cells and their response to chemotherapeutic drugs.METHODS:A 1.03kb cDNA sequence of Hsp90beta was obtained from the primary plasmid phHSP90 by EcoR I and BamH I nuclease digestion and was cloned to the EcoR I and BamH I site of the pcDNA by T4DNA ligase and an antisense orientation of Hsp90beta expression vector was constructed. The constructs were transfected with lipofectamine and positive clones were selected with G418. The expression of RNA was determined with dot blotting and RNase protection assay, and the expression of Hsp90 protein determined with western blot. Cell cycle distribution of the transfectants was analyzed with flow cytometry, and the drug sensitivity of the transfectants to Adriamycin (ADR), vincrinstine (VCR), mitomycin (MMC) and cyclophosphamide (CTX) with MTT and intracellular drug concentration of the transfectants was determined with flow cytometry.RESULTS:In EcoR I and BamH I restriction analysis, the size and the direction of the cloned sequence of Hsp90beta remained what had been designed and the gene constructs were named pcDNA-Hsp90.AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 cell clones all expressed Hsp90 anti-sense RNA. The expression of Hsp90 was down-regulated in AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 cell clones. Cell cycle distribution was changed differently. In AH-SGC7901/VCR and AH-Ec109 cells, G(1) phase cells were increased; S phase and G(2) phase cells were decreased as compared with their parental cell lines. In AH-SGC7901 cell, G(1)phase cells were decreased, G(2) phase cells increased and S phase cells were not changed, and in AH-HCC7402 cells G(1), S and G(2) phase cells remained unchanged as compared with their parental cell lines. The sensitivity of AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 to chemotherapeutic drugs, the sensitivity of AH-SGC7901/VCR to ADR, VCR, MMC and CTX the sensitivity of AH-HCC7402 to ADR and VCR, and the sensitivity of Ec109 to ADR, VCR and CTX all increased as compared with their parental cell lines. The mean fluorescence intensity of ADR in AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 was also significantly elevated (P 【 0.05).CONCLUSION: Down-regulation of Hsp90 could change cell cycle distribution and increase the drug sensitivity of tumor cells.

Devonian Sequence Stratigraphy and Sea Level Change Cycles in South China

The Devonian is well developed in South China and has drawn a great attention from the geologists both at domestic and abroad. On the basis of study on the sequence stratigraphy in more than 10 sections in Guizhou, Guangxi, Longmenshan and Southern Qinling within South China we have identified 21 sequences and T-R cycles that correspond to third-order sea level changes. These sea level change cycles were controlled by autorhythm(Pragian,Eifelian), allorhythm(Lochkovian,Emsian,Givetian) and coupling rhythm mechanisms (frasian, Famennian and F-F boundary).

Modulating effects of survivin antisense oligonucleotide on changes of apoptosis and cell cycle of human hepatocellular carcinoma cell line SMMC-7721

To investigate the modulating effects of survivn antisense oligonucletode (ASODN) on the cell cycle and apoptosis of human hepatocellular carcinoma (HCC) cell line SMMC-7721 and explore its mechanism.Methods Survivin ASODN was transfected into SMMC-7721 cells mediated by DOTAP liposomal reagent.Electron microscopy,flow cytometry and RT-PCR were used to detect the changes in cell ultrastructure,apoptosis,cell cycle and the expression of cyclinB1 mRNA,respectively.Results After transfection of survivin ASODN,the expression of cyclinB1 mRNA in the cells significantly increased and increase in G2-M arrest and apoptosis appeared.Meanwhile,the cell ultrastructure had apoptotic changes such as chromatin condensation and apoptotic body formation.Conclusion Survivin ASODN can induce the expression of cyclinB1 that may result in G2-M arrest.Consequently,apoptosis is triggered.Survivin ASODN transfection might be an improtant new treatment for HCC.14 refs,2 figs,1 tab.

Three-step modernization of the ocean:Modeling of carbon cycles and the revolution of ecological systems in the Ediacaran/Cambrian periods

Important ecological changes of the Earth （oxidization of the atmosphere and the ocean） increase in nutrient supply due to the break-up of the super continent （Rodinia） and the appearance of multi-cellular organisms （macroscopic algae and metazoan） took place in the Ediacaran period, priming the Cambrian explosion. The strong perturbations in carbon cycles in the ocean are recorded as excursions in carbonate and organic carbon isotope ratio （δ13Ccarb and δ13Corg） from the Ediacaran through early Cambrian periods. The Ediacaran-early Cambrian sediment records of δ13Ccarb and δ13Corg, obtained from the drill-core samples in Three Gorges in South China, are compared with the results of numerical simulation of a sim- ple one-zone model of the carbon cycle of the ocean, which has two reservoirs （i.e., dissolved organic carbon （DOC） and dissolved inorganic carbon （DIC）. The fluxes from the reservoirs are assumed to be proportional to the mass of the carbon reservoirs. We constructed a model, referred to here as the Best Fit Model （BFM）, which reproduce δ13Ccarb and δ13Corg records in the Ediacaran-early Cambrian period noted above. BFM reveals that the Shuram excursion is related to three major changes in the carbon cycle or the global ecological system of the Earth： （1） an increase in the coefficient of remineralization by a factor of ca. 100, possibly corresponding to a change in the dominant metabolism from anaerobic respiration to aerobic respiration, （2） an increase of carbon fractionation index from 25‰, to 33‰, possibly corresponding to the change in the primary producer from rock-living cyanobacteria to free-living macro algae, and （3） an in- crease in the coefficient of the organic carbon burial by a factor of ca. 100, possibly corresponding to the onset of a biological pump driven by the flourishing metazoan and zooplankton. The former two changes took place at the start of the Shuram excursion, while the third occurred at the end of the Shuram excursion. The other two excursions are explained by the tentative decrease in primary production due to cold periods, which correspond to the Gaskiers （ca. 580 Ma） and Bikonor （ca. 542 Ma） glaciations.

Triassic Sequence Stratigraphy and Sea-Level Changes in Qomolongma Area, Southern Tibet, China: From Epicontinental Sea to Rift Basin

The Triassic in the Qomolongma area, southern Tibet, was deposited under an extensional tectonic setting from the Pangea supercontinent to continental rifting. From the Induan to Rhaetian, 12 depositional sequences (3rd order) have been recognized, which can be grouped into 5 sequence sets and in turn make up a well defined mesosequence (2nd order). Among the recognized marine transgressions, those at 250 Ma, 239 Ma, 231 Ma and 223 Ma respectively are particularly of significance and can be correlated widely across continents. The study shows that in Triassic the Qomolongma area experienced a sedimentary evolution from epicontinental sea to rift basin with the turning point at ca 228 Ma. During the early and middle epochs, the area was under epeiric sea, with carbonate ramp to mixed shoal environments predominant. In the late Carnian, the strong extension initiated listric faulting, thus resulting in rapid basement subsidence and the onset of a rift basin. From the late Norian to Rhaetian, it manifested as a rapid basin filling process in the area. Coupled with long term sea level fall, the excessive terrigenous influx led to the shift of environment from deep water prodelta to shore and finally to fluvial plain.

Cambrian Sequence Stratigraphy and Sea Level Cycles of North China Platform

The Cambrian of the North China platform consists chiefly of shallow water deposits and shows the sedimentary characters of an epicontinental sea basin. Controlled mainly by global sea level changes and sedimentary influx, the depositional sequences all exhibit as composite sequences. From bottom upward, 14 sequences (3rd order) are recognized, which may be grouped into 5 sequence sets and further into 2 mesosequences (2nd order). It is suggested herein that the Cambrian/Ordovician boundary may better be set at the MFS (maximum flooding surface) of the sequence OSq1, above which the conodont Cordylodus lindstroemi occurs. This position is about 40 m above the traditional Cambrian/Ordovician boundary and is within the Yeli Formation.

CYCLE OF CHANGE

While huge teams of bicyclists are no longer a common scene in China, long lines of cars can be seen everywhere. But does this mean that bicycles will cease to be a major mode of transport in the country?

Assessment of the Endothelial Function with Changed Volume of Brachial Artery by Menstrual Cycle

The endothelial function has been proven to be an important factor in the pathogenesis of atheroscle-rosis, hypertension and heart failure. The flow-mediated vasodilation (FMD) of the peripheral artery is an endothelium-dependent function. Brachial-artery ultrasound scanning is the popular method for evaluat-ing FMD.

Diurnal and spatial variations of soil NO_x fluxes in the northern steppe of China

NOx emissions from biogenic sources in soils play a significant role in the gaseous loss of soil nitrogen and consequent changes in tropospheric chemistry. In order to investigate the characteristics of NOx fluxes and factors influencing these fluxes in degraded sandy grasslands in northern China, diurnal and spatial variations of NOx fluxes were measured in situ. A dynamic flux chamber method was used at eight sites with various vegetation coverages and soil types in the northern steppe of China in the summer season of 2010.Fluxes of NOx from soils with plant covers were generally higher than those in the corresponding bare vegetation-free soils, indicating that the canopy plays an important role in the exchange of NOx between soil and air. The fluxes of NOx increased in the daytime,and decreased during the nighttime, with peak emissions occurring between 12：00 and14：00. The results of multiple linear regression analysis indicated that the diurnal variation of NOx fluxes was positively correlated with soil temperature（P 〈 0.05） and negatively with soil moisture content（P 〈 0.05）. Based on measurement over a season, the overall variation in NOx flux was lower than that of soil nitrogen contents, suggesting that the gaseous loss of N from the grasslands of northern China was not a significant contributor to the high C/N in the northern steppe of China. The concentration of NOx emitted from soils in the region did not exceed the 1-hr National Ambient Air Quality Standard（0.25 mg/m^3）.

Winter survival of microbial contaminants in soil:An in situ verification

The aim of the research was to evaluate, at site scale, the influence of freezing and freeze/thaw cycles on the survival of faecal coliforms and faecal enterococci in soil, in a climate change perspective. Before the winter period and during grazing, viable cells of faecal coliforms and faecal enterococci were detected only in the first 10 cm below ground, while,after the winter period and before the new seasonal grazing, a lower number of viable cells of both faecal indicators was detected only in some of the investigated soil profiles, and within the first 5 cm. Taking into consideration the results of specific investigations, we hypothesise that the non-uniform spatial distribution of grass roots within the studied soil can play an important role in influencing this phenomenon, while several abiotic factors do not play any significant role. Taking into account the local trend in the increase of air temperature, a different distribution of microbial pollution over time is expected in spring waters, in future climate scenarios. The progressive increase in air temperature will cause a progressive decrease in freeze/thaw cycles at higher altitudes, minimising cold shocks on microbial cells, and causing spring water pollution also during winter.