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In order to know about the rheological properties of typical biotite granitic gneiss in Xiaowan Hydropower Project, triaxial rheological experiments with biotite granitic gneiss were carried out on the rock servo-controlling rheological testing machine. As a result the typical complete rheological curve of the biotite granitic gneiss indicates that when the stress level is low, the rheological deformation of rock specimen is not obvious; when in high stress conditions, the rock mass rheological properties are very significant with large rheological deformation, including three typical rheological phases. The rheological deformation decreases with the increase of the confining pressure, and at the same time the rheological rate of rock specimen changes with the change of stress level. In low stress state, the corresponding rheological rate of the rock specimen incarnated as the rheological rate attenuation stage and the rheological rate uniform stage, while once the stress level exceeds the long-term strength of rock specimen, the rheological rate would first decreases and then keep at a constant value, and finally it will trend to follow the non-linear acceleration law. The rupture form of the low-intensity biotite granitic gneiss specimen is the main crack surface which causes the damage of rock specimen, and finally generated by the expansion and transfixion of the crack in the initial cavern flaw. The rheological deformation and rupture form of rock specimen in different confining pressures are not exactly the same.

Rheological experiments were carried out for sandstone and marble specimens from left bank high slope of Jingping First Stage Hydropower Project by using the rock servo-controlling rheology testing machine. Typical triaxial rheological curves under step loading and temperature curves in the process of rheological experiment were gained. BP neural network is improved by Levenberg-Marquardt algorithm. Improved neural network model for rock rheology is established in accordance with the rheology experimental results of rock specimen. The improved neural network model was used to forecast rock rheological experimental curves, and the result shows that the forecasted rock rheology curves are closely accorded with the experimental result. The improved neural network model takes into account the influence of loading history and temperature difference on the rock rheological deformation, and the forecasted result can reflect better the rheology deformation behavior of rock material.

Based on the results of triaxial unloading tests，the stress-strain characteristics and failure characteristics of rock mass have been discussed. Considering unloading degree as an important parameter，an elastic-brittle-plastic mechanical model has been established according to the unloading stress condition，which considers that unloading yield function varies linearly between Griffith with Hoek-Brown criterion after rock mass receives plastic deformation caused by unloading. Taking jinshajiang excavation slope of a hydropower station as engineering background, the three-dimensional continua fast Lagrangian analysis software FLAC3D is employed in the slope stability analysis under the excavation, and the distribution characteristics of the slop deformation is studied.

According to the specific geological and topographical conditions at slope of the Hydropower Station, considering the shape and size of simulation geometric model, the position relationship between the simulated area and buildings of terminal area and boundary conditions, the dynamic model which is fit to the geological structure are selected. Using the measured stress distribution as constrained conditions, 3D initial geostress field of research area are simulated using FLAC3D program. The result of simulation analysis shows that 3D initial geostress field of research area are accurately simulated based on good combination between the measured stress data and numeriacal simulation. So as to provide a reasonable basis for further condition simulation, such as excavation and unloading conditions. It has important significance for similar geological environment projects that the method is successfully applied to simulate initial geostress of this area.

Expression of the breast cancer-associated gene 1 (BRCA1) in sporadic breast cancers is usually reduced, yet the underlying mechanisms remains elusive. To identify factors that are responsible for reduced BRCA1 expression, we screened 92 known transcription factors for their ability to regulate expression of BRCA1. Among several potential regulators, the Gli-Krueppel-related transcription factor Yin Yang 1 (YY1) showed the most dramatic transactivation of the BRCA1 promoter. YY1 binds to the promoter of BRCA1, and its overexpression resulted in increased expression of BRCA1 and a number of BRCA1 downstream genes. We further showed that overexpression of YY1 in cancer cells inhibited cell proliferation, foci formation and tumor growth in nude mice. To assess the clinical relevance between YY1 and BRCA1, we studied expression of YY1 and BRCA1 from human breast cancer samples and tissue arrays, and detected a significant positive correlation between the level of YY1 and BRCA1 expression in these cancers. Taken together, these findings suggest that YY1 is a key regulator of BRCA1 expression and may be causally linked to the molecular etiology of human breast cancer.

During the hydraulic structure design, the stability of abutment slope is critical to the normal operation of hydraulic structure. Through the tests, the effects of different hydraulic pressure on the strength and deformation characteristics of sandstone were studied, and the variation characteristics of rock masses with joints under secondary loading after excavation damage were analyzed as well. Test results show that: 1. When the samples without immersion secondly load until complete failure, ultimate strain increases slightly than the first loading, of which the increasing value is 7.5%; elastic modulus has a relatively large reduction than the first, which reduces 22.8%; deformation modulus has a less reduction, which is about 3.3%; 2. When considering the softening action of different hydraulic pressure on samples, ultimate strain increases by about 23% than that without consideration of immersion effect; elastic modulus reduces about 53%; deformation modulus reduces about 39%; 3. With the increment of immersion pressure, sample strength decreases greater, so does the softening effect of water on rock samples. With the increment of pressure, the strength of rock with joints has a relatively large reduction, which is from 27% to 41%, and the increasing value of intact rock mass isn't great, which is only from 25% to 27%. Jointed rock masses are more sensitive to the softening effect of water, and mechanical properties weaken obviously after long-term immersion with unstable problem.

For geotechnical engineering, it has great significance to research the mechanical characteristics of rock mass under three dimensional stresses. Through triaxial compression failure test, the characteristics of stress-strain curve and deformation of red sandstone from Sichuan under different confining pressures has been analyzed. The results show that: with the increment of confining pressure, the failure mode of rock mass changes from brittle failure to ductile failure gradually, and an obvious yield platform appears near the peak strength of stress-strain curve; the elastic modulus, deformation modulus, peak strain and residual strain of rock sample increase with the increment of confining pressure, the elastic modulus and deformation modulus are not a fixed value, and the relation between deformation parameter and confining pressure can be fit through quadratic curve.

The molybdenum alloy sheets composite strengthened by silicon and lanthanum oxide were prepared by powder metallurgy technology with Mo-La2O3(0.3wt%) and Si(0, 0.1, 0.3wt%) powders and thermo-mechanically processing. The influences of silicon content on the microstructure and mechanical properties of the final molybdenum alloy sheets were tested and analysized. The results show that the addition of lanthanum oxide and silicon can refine the alloys grain size. The introduction of lanthanum oxide particles can increase the yield strength. Although the molybdenum alloys with 0.3wt% silicon have solid solution strengthening effect, the alloys with 0.1 wt% silicon exhibits obvious solid solution softening effect at room temperature. The strengthening mechanisms are quantitatively assessed, which well explain the increase or decrease in yield strength with respect to grain size, lanthanum oxide particle and silicon solid solution.

Rock rheology is common in engineering practice, which has an effect on long-term security and stability of the project. In order to understand the rheological mechanical properties under the condition of unloading sand, automatic servo apparatus of triaxial creep of rock was used to have a rheological mechanical unloading triaxial test on sandstone, which got typical flow complete curves of rock. Rheological test results showed that rheological phenomena occured in sandstone, and its rheological properties were significant with large rheological deformation, especially in high-stress conditions, When the external load exceeded the long-term strength of rock samples, specimen deformation went through three typical rheological stages with time increasing, and ultimately speed up the flow breakdown. Based on the experimental results, analyzed and studied the rheological parameters that were obtained, and discussed unsteady character law of rheological parameters. defined rheological damage of rock in the accelerating flow process as the unloading capacity and time-related exponential function, and proposed corresponding damage evolution equation, brought damage evolution equation into Burgers rheological model, and obtained nonlinear unloading model of rheological damage of rock, used accelerating rheological test curve of sandstone samples under the condition of 15MPa confining pressure and 135MPa stress level to verify nonlinear unloading model of rheological damage of rock, and the results showed that the fitting results of rheological test data and the improved rheological model had good consistency, and the rheological model can reflect the three stages of nonlinear rheology of rock, which can reflect the rheological characteristics under the condition of unloading sandstone more accurately.

Most of the engineering rock masses contain a variety of different levels of geological tectonic joints and weak planes, which can weaken the rock strength. The rock masses containing joints have completely different mechanical properties with the intact ones. Through loading failure tests on the rock masses containing two intermittent joints of prefabricated of different spacing, the differences between jointed rock mass and intact one were studied. The research shows that: 1. Comparing with the intact rock mass, the stress-strain curve of jointed one has a relatively large fluctuation near the peak, it isn’t smooth, and there's a reduction in the stage of plastic flow after yielding; ultimate strength decreases obviously, joint depth has a great impact on strength, and there's no necessary link between ultimate strength of rock mass and joints spacing. 2. When the loading is failure, the elastic and deformation modulus of rock mass decrease obviously comparing with those of the intact rock mass, which tend small generally with the increment of joints spacing, however, they have a relatively complex relation and it isn't linear. 3. The failure characteristics of jointed rock mass are different from those of the intact rock mass, failure planes are relatively complex and no longer single shear or complementary shear ones, which presents that shear failures occur along the end of prefabricated joints with few extensional cracks; the spacing of prefabricated joints have a great impact on the failure pattern of rock mass. The research results can provide certain references for the mechanical parameters selection of jointed rock mass of engineering design and numerical analysis.

The brittle is crippling the application of bioceramic. The compound bioceramic is a new biomaterial being widely applied in medical treatments and its fracture toughness is an important mechanical behaviors. In this paper, we introduce the manufacturing method of the compound bioceramic and experiment facilities for its fracture toughness, investigate its probability distribution for the experimental data and conduct the test for fit. We conclude that the experimental data for the toughness fracture of the compound bioceramic obey the two-parameter Weibull distribution, introduce the analyzing method for the upper confidence limit curve and lower confidence limit curve and study the reliability and confidence level of the fracture toughness of the compound bioceramic.

Resveratrol (RSV) is a naturally occurring polyphenol that has been found to exert antioxidant, anti-inflammatory, and neuroprotective properties. However, how RSV exerts its beneficial health effects remains largely unknown. Here, we show that RSV inhibits insulin- and leucine-stimulated mTOR signaling in C2C12 fibroblasts via a Sirt1-independent mechanism. Treating C2C12 cells with RSV dramatically inhibited insulin-stimulated Akt, S6 kinase, and 4E-BP1 phosphorylation but had little effect on tyrosine phosphorylation of the insulin receptor and activation of the p44/42 MAPK signaling pathway. RSV treatment also partially blocked mTOR and S6 kinase phosphorylation in TSC1/2-deficient mouse embryonic fibroblasts, suggesting the presence of an inhibitory site downstream of TSC1/2. Knocking out PDK1 or suppressing AMP-activated protein kinase had little effect on leucine-stimulated mTOR signaling. On the other hand, RSV significantly increased the association between mTOR and its inhibitor, DEPTOR. Furthermore, the inhibitory effect of RSV on leucine-stimulated mTOR signaling was greatly reduced in cells in which the expression levels of DEPTOR were suppressed by RNAi. Taken together, our studies reveal that RSV inhibits leucine-stimulated mTORC1 activation by promoting mTOR/DEPTOR interaction and thus uncover a novel mechanism by which RSV negatively regulates mTOR activity.

Shot peening’s surface roughness is an important factor affecting the effect of shot-peening. The paper selects blasting pressure, scanning speed and target distance as affecting factors in the process parameters, the shot penning test which aims at 2A11 aluminum alloy materials through applying the premixed water jet, according to test data, the paper establishes mathematical model of shot peening’s surface roughness applying neural network, and applies this model to predict shot peening’s surface roughness. The results show that the training average error of this model is small, the predicted effect is good, it can meet the requirements of shot peening’s surface roughness prediction accuracy in the industrial production, it has greater practical value.

Global DNA hypomethylation at CpG islands coupled with local hypermethylation is a hallmark for breast cancer, yet the mechanism underlying this change remains elusive. In this study, we showed that DNMT1, which encodes a methylation maintenance enzyme, is a transcriptional target of BRCA1. BRCA1 binds to the promoter of the DNMT1 gene through a potential OCT1 site and the binding is required for maintaining a transcriptional active configuration of the promoter in both mouse and human cells. We further demonstrated that impaired function of BRCA1 leads to global DNA hypomethylation, loss of genomic imprinting, and an open chromatin configuration in several types of tissues examined in a BRCA1 mutant mouse model at premaligant stages. BRCA1 deficiency is also associated with significantly increased expression levels of several protooncogenes, including c-Fos, Ha-Ras, and c-Myc, with a higher expression in tumors, while premalignant mammary epithelial cells displayed an intermediate state between tumors and controls. In human clinical samples, reduced expression of BRCA1 correlates with decreased levels of DNMT1, and reduced methylation of CpG islands. Thus, BRCA1 prevents global DNA hypomethylation through positively regulating DNMT1 expression, and this provides one of mechanisms for BRCA1-associated breast cancer formation.

SummaryUnder various conditions, mammals have the ability to maintain serum glucose concentration within a narrow range. SIRT1 plays an important role in regulating gluconeogenesis and fat metabolism; however, the underlying mechanisms remain elusive. Here, we show that SIRT1 forms a complex with FOXO3a and NRF1 on the SIRT6 promoter and positively regulates expression of SIRT6, which, in turn, negatively regulates glycolysis, triglyceride synthesis, and fat metabolism by deacetylating histone H3 lysine 9 in the promoter of many genes involved in these processes. Liver-specific deletion of SIRT6 in mice causes profound alterations in gene expression, leading to increased glycolysis, triglyceride synthesis, reduced β oxidation, and fatty liver formation. Human fatty liver samples exhibited significantly lower levels of SIRT6 than did normal controls. Thus, SIRT6 plays a critical role in fat metabolism and may serve as a therapeutic target for treating fatty liver disease, the most common cause of liver dysfunction in humans.

A new species, Scrophularia jinii (Scrophulariaceae), from Central China is described and illustrated. This new species was formerly misidentified as S. fargesii, from which it differs in many morphological characters. Moreover, it is distinct with all known Scrophularia species in its unique deeply double serrate leaf margin with 3–7 big teeth on each side. Molecular phylogenetic analysis further supports its species delimitation and suggests a close relationship with several Japanese and North American species.

Checkpoint kinase 1 (Chk1) is a key element in the DNA-damage response pathway that is required for maintaining genomic stability. To study the potential role of Chk1 in mammary tumorigenesis, we disrupted it using a Cre/loxP system. We showed that although Chk1 heterozygosity caused abnormal development of the mammary gland, it was not sufficient to induce tumorigenesis. Simultaneous deletion of one copy of p53 failed to rescue the developmental defects; however, it synergistically induced mammary tumor formation in Chk1(+/−); MMTV-Cre animals with a median time to tumor latency of about 10 months. Chk1 deficiency caused a preponderance of abnormalities, including prolongation, multipolarity, misalignment, mitotic catastrophe and loss of spindle checkpoint, that are accompanied by reduced expression of several cell cycle regulators, including Mad2. On the other hand, we also showed that Chk1 deficiency inhibited mammary tumor formation in mice carrying a homozygous deletion of p53, uncovering a complex relationship between Chk1 and p53. Furthermore, inhibition of Chk1 with a specific inhibitor, SB-218078, or acute deletion of Chk1 using small hairpin RNA killed mammary tumor cells effectively. These data show that Chk1 is critical for maintaining genome integrity and serves as a double-edged sword for cancer: although its inhibition kills cancer cells, it also triggers tumorigenesis when favorable mutations are accumulated for cell growth.

The tumor suppressor BRCA1 interacts with many proteins and undergoes multiple modifications on DNA damage. ATM, a key molecule of the DNA damage response, phosphorylates S1189 of BRCA1 after γ-irradiation. S1189 of BRCA1 is known as a unique ATM phosphorylation site in BRCA1 exon 11. To study the functions of ATM-dependent phosphorylation of BRCA1-S1189, we generated a mouse model carrying a mutation of S1152A (S1152 in mouse Brca1 corresponds to S1189 in human BRCA1) by gene targeting. Brca1S1152A/S1152A mice were born at the expected ratio, unlike that seen in previous studies of Brca1-null mice. However, 36% of Brca1S1152A/S1152A mice exhibited aging-like phenotypes including growth retardation, skin abnormalities, and delay of the mammary gland morphogenesis, with an increase in apoptosis. Mutant mice were hypersensitive to high doses of γ-irradiation, displaying shortened life span and reduction in intestinal villus size, associated with increased apoptosis. Aging-unaffected 18-month-old Brca1S1152A/S1152A female mice also showed mammary gland abnormalities with increased levels of cyclin D1 and phospho-ER-α, such as Brca1-Δ11 mutation. On low-dose γ-irradiation, they suffered a marked increase in tumor formation with an abnormal coat pattern. Furthermore, Brca1S1152A/S1152A embryonic fibroblasts failed to accumulate p53 on γ-irradiation with delayed phosphorylation of p53-S23. These observations indicate that ATM-mediated phosphorylation of S1189 is required for BRCA1 functions in the modulation of DNA damage response and in the suppression of tumor formation by regulating p53 and apoptosis. [Cancer Res 2009;69(24):9291–300]

Polypropylene synthetic paper releasing anion was prepared from polypropylene resin, anion additives, titanium dioxide, etc., by calendar forming method. The synthetic paper was tested by anion detector, SEM, AFM, etc. Tensile strength, elongation at break, right angle tear strength of the polypropylene synthetic paper reached the GB 13022 or QB/T1130 Standard. The synthetic paper was water and oil resistance, and released anions 10,530 cm −3 . It was environment-friendly, and a kind of good material for human's health.

Almost all complex multicellular organisms on earth utilize oxygen for the production of energy. This strategy carries the risk for damaging ROS to be generated and so these biochemical pathways must be highly regulated. Because of this, regulation of oxidative-phosphorylation is tightly coordinated with every aspect of cellular physiology, including stem cell regulation during embryonic development and in adult organisms. The protein-deacetylase, SIRT1, has received much attention because of its roles in oxygen metabolism, cellular stress response, aging, and has been investigated in various species and cell types including embryonic stem cells. However, there is a dearth of information on SIRT1 in adult stem cells, which have a pivotal role in adult aging processes. Here, we discuss the potential relationships between SIRT1 and the surface receptor protein, Notch, with stem cell self-renewal, asymmetric cell division, signaling and stem cell aging.

It is hypothesized that cancer stem cells arise either from normal stem cells or from progenitor cells that have gained the ability to self-renew. Here we determine whether mammary cancer stem cells can be isolated by using antibodies that have been used for the isolation of normal mammary stem cells. We show that BRCA1 mutant cancer cell lines contained a subpopulation of CD24+CD29+ or CD24+CD49f+ cells that exhibited increased proliferation and colony forming ability in vitro, and enhanced tumor-forming ability in vivo. The purified CD24+CD29+ cells could differentiate and reconstitute the heterogeneity found in parental cells when plated as a monolayer. Under low-attachment conditions, we detected “tumorspheres” only in the presence of double positive cells, which maintained their ability to self-renew. Furthermore, CD24+CD29+ cells could form tubular structures reminiscent of the mammary ductal tree when grown in three-dimensional cultures, implying that these cancer cells maintain some of the characteristics of the normal stem cells. Nevertheless, they could still drive tumor formation since as low as 500 double positive cells immediately after sorting from BRCA1 mutant primary tumors were able to form tumors with the same heterogeneity found in the original tumors. These data provide evidence that breast cancer stem cells originate from normal stem cells and advance our understanding of BRCA1-associated tumorigenesis with possible implications for future cancer treatment.

In thalassemia, deficient globin-chain production during erythropoiesis results in anemia. Thalassemia may be further complicated by iron overload (frequently exacerbated by blood transfusion), which induces numerous endocrine diseases, hepatic cirrhosis, cardiac failure and even death. Accumulation of iron in the absence of blood transfusions may result from inappropriate suppression of the iron-regulating peptide hepcidin by an erythropoietic mechanism. To test this hypothesis, we examined erythroblast transcriptome profiles from 15 healthy, nonthalassemic donors. Growth differentiation factor 15 (GDF15), a member of the transforming growth factor-beta superfamily, showed increased expression and secretion during erythroblast maturation. Healthy volunteers had mean GDF15 serum concentrations of 450 +/- 50 pg/ml. In comparison, individuals with beta-thalassemia syndromes had elevated GDF15 serum levels (mean 66,000 +/- 9,600 pg/ml; range 4,800-248,000 pg/ml; P < 0.05) that were positively correlated with the levels of soluble transferrin receptor, erythropoietin and ferritin. Serum from thalassemia patients suppressed hepcidin mRNA expression in primary human hepatocytes, and depletion of GDF15 reversed hepcidin suppression. These results suggest that GDF15 overexpression arising from an expanded erythroid compartment contributes to iron overload in thalassemia syndromes by inhibiting hepcidin expression.

To summarize and analyze neonatal screening results for congenital hypothyroidism (CH) in parts of Yunnan Province, China.A total of 236 218 newborns (121 463 males and 114 755 females) who were born in Zhaotong City, Qujing City, Lijiang City, and Diqing Tibetan Autonomous Prefecture of Yunnan Province, China, between July 2012 and April 2014 were screened for CH. The original blood smear was re-tested if the thyroid stimulating hormone (TSH) level in heel blood was ≥8 μIU/L in the initial screening. The newborns with positive TSH results were called back for further diagnosis by measuring blood TSH and free thyroxine (FT4) levels.Among 236 218 newborns, the pass rate of blood smears, re-acquisition rate of unqualified blood smears, and recall rate of suspected cases were 96.67%, 81.75%, and 73.02%, respectively. Sixty-six cases of CH were confirmed, among which 36 were male infants and 30 were female infants (P0.05). The incidence rate of CH was 1/3 579, which was significantly lower than the national average rate (1/2 034; P0.01). The gestational age of CH newborns was mostly between 37 to 42 weeks, and only 3% were born at a gestational age of42 weeks. Most of the CH newborns had normal birth weight. The CH newborns with a body length of50 cm accounted for 32%.The incidence of CH in Yunnan Province is lower than the national average. There are no specific clinical features in CH newborns. The neonatal screening in Yunnan Province needs further improvement.

Cholangiocellular carcinoma (CC), the second most common primary liver cancer, is associated with a poor prognosis. It has been shown that CCs harbor alterations of a number of tumor-suppressor genes and oncogenes, yet key regulators for tumorigenesis remain unknown. Here we have generated a mouse model that develops CC with high penetrance using liver-specific targeted disruption of tumor suppressors SMAD4 and PTEN. In the absence of SMAD4 and PTEN, hyperplastic foci emerge exclusively from bile ducts of mutant mice at 2 months of age and continue to grow, leading to tumor formation in all animals at 4-7 months of age. We show that CC formation follows a multistep progression of histopathological changes that are associated with significant alterations, including increased levels of phosphorylated AKT, FOXO1, GSK-3beta, mTOR, and ERK and increased nuclear levels of cyclin D1. We further demonstrate that SMAD4 and PTEN regulate each other through a novel feedback mechanism to maintain an expression balance and synergistically repress CC formation. Finally, our analysis of human CC detected PTEN inactivation in a majority of p-AKT-positive CCs, while about half also lost SMAD4 expression. These findings elucidate the relationship between SMAD4 and PTEN and extend our understanding of CC formation.

Wee1 kinase regulates the G2/M cell cycle checkpoint by phosphorylating and inactivating the mitotic cyclin-dependent kinase 1 (Cdk1). Loss of Wee1 in many systems, including yeast and drosophila, leads to premature mitotic entry. However, the developmental role of Wee1 in mammals remains unclear. In this study, we established Wee1 knockout mice by gene targeting. We found that Wee-/- embryos were defective in the G2/M cell cycle checkpoint induced by γ-irradiation and died of apoptosis before embryonic (E) day 3.5. To study the function of Wee1 further, we have developed MEF cells in which Wee1 is disrupted by a tamoxifen inducible Cre-LoxP approach. We found that acute deletion of Wee1 resulted in profound growth defects and cell death. Wee1 deficient cells displayed chromosome aneuploidy and DNA damage as revealed by γ-H2AX foci formation and Chk2 activation. Further studies revealed a conserved mechanism of Wee1 in regulating mitotic entry and the G2/M checkpoint compared with other lower organisms. These data provide in vivo evidence that mammalian Wee1 plays a critical role in maintaining genome integrity and is essential for embryonic survival at the pre-implantation stage of mouse development.

Cyclin-dependent kinase 2 (Cdk2) phosphorylates Thr320 of protein phosphatase 1alpha (PP1alpha) in late G(1), thereby inhibiting its activity. Phosphorylation-resistant PP1alphaT320A, acting as a constitutively active (CA) mutant, causes a late G(1) arrest by preventing the phosphorylation and inactivation of the retinoblastoma protein (pRb). Both PP1alpha-mediated G(1) arrest and PP1alpha phosphorylation in late G(1) require the presence of pRb, indicating that PP1alpha is a crucial regulator of the pRb pathway, which is almost invariably mutated in human cancer. These findings prompted us to investigate whether PP1alpha interferes with oncogenic transformation. The ability of NIH 3T3 cells to form foci after transformation with ras/cyclin D1 was significantly inhibited by co-transfection with PP1alphaT320A, but not PP1alpha. Likewise, cells expressing PP1alphaT320A or PP1alphaT320A fused to green fluorescent protein (GFP) were unable to form colonies in soft agar, regardless of whether PP1alpha constructs were co-transfected with ras/cyclin D1 or transfected into stably transformed cells. Overexpressed wild-type (Wt) PP1alpha and GFP-PP1alpha were phosphorylated in Thr320, most likely explaining its lack of effect. Expression of GFP-PP1alphaT320A was associated with caspase-cleaved pRb in Western blots (WB) and morphological signs of cell death. These findings demonstrate that PP1alpha activity can override oncogenic signaling by causing cell-cycle arrest and/or apoptosis rather than restoring contact inhibition or anchorage dependence.

BRCA1 and BRCA2 mutations are responsible for most familial breast carcinomas. Recent reports carried out in non-cancerous mouse BRCA1- or BRCA2-deficient embryonic stem (ES) cells, and hamster BRCA2-deficient cells have demonstrated that the targeted inhibition of poly(ADP-ribose) polymerase (PARP-1) kills BRCA mutant cells with high specificity. Although these studies bring hope for BRCA mutation carriers, the effectiveness of PARP-1 inhibitors for breast cancer remains elusive. Here we present the first in vivo demonstration of PARP-1 activity in BRCA1-deficient mammary tumors and describe the effects of PARP-1 inhibitors (AG14361, NU1025, and 3-aminobenzamide) on BRCA1-deficient ES cells, mouse and human breast cancer cells. AG14361 was highly selective for BRCA1-/- ES cells; however, NU1025 and 3-aminobenzamide were relatively non-selective. In allografts of naïve ES BRCA1-/- cells there was either partial or complete remission of tumors. However, in allografts of mouse, BRCA1-/- mammary tumors, there was no tumor regression or remission although a partial inhibition of tumor growth was observed in both the BRCA1-/- and BRCA1+/+ allografts. In human tumor cells, PARP-1 inhibitors showed no difference in vitro in limiting the growth of mammary tumors irrespective of their BRCA1 status. These results suggest that PARP-1 inhibitors may non-specifically inhibit the growth of mammary tumors.

In mammalian spermatogenesis, the X and Y chromosomes are transcriptionally silenced during the pachytene stage of meiotic prophase (meiotic sex chromosome inactivation, MSCI), forming a condensed chromatin domain termed the sex or XY body [1–3]. The nucleosomal core histone H2AX is phosphorylated within the XY chromatin domain just prior to MSCI, and it has been hypothesized that this triggers the chromatin condensation and transcriptional repression [4, 5]. Here, we show that the kinase ATR localizes to XY chromatin at the onset of MSCI and that this localization is disrupted in mice with a mutant form of the tumor suppressor protein BRCA1. In the mutant pachytene cells, ATR is usually present at nonsex chromosomal sites, where it colocalizes with aberrant sites of H2AX phosphorylation; in these cells, there is MSCI failure. In rare pachytene cells, ATR does locate to XY chromatin, H2AX is then phosphorylated, a sex body forms, and MSCI ensues. These observations highlight an important role for BRCA1 in recruiting the kinase ATR to XY chromatin at the onset of MSCI and provide compelling evidence that it is ATR that phosphorylates H2AX and triggers MSCI.

BRCA1-associated breast cancer exhibits significantly higher levels of chromosomal abnormalities than sporadic breast cancers. However, the molecular mechanisms regarding the roles of BRCA1 in maintaining genome integrity remain elusive. By using a mouse model deficient for Brca1 full-length isoform ( Brca1 Δ 11/ Δ 11 ), we found that Brca1 Δ 11/ Δ 11 cells displayed decreased expression of a number of genes that are involved in the spindle checkpoint, including Mad2, which is a key component of spindle checkpoint that inhibits anaphase-promoting complex. We showed that Brca1 Δ 11/ Δ 11 cells failed to arrest at metaphase in the presence of nocodazole and underwent apoptosis because of activation of p53. Consistently, reconstitution of Mad2 in Brca1 Δ 11/ Δ 11 cells partially restored the spindle checkpoint and attenuated apoptosis. By using UBR60 cells, which carry tetracycline-regulated expression of BRCA1, we demonstrated that BRCA1 binds to transcription factor OCT-1 and up-regulates the transcription of MAD2. Furthermore, we showed that the induction of BRCA1 to endogenous MAD2 or transfected MAD2 luciferase reporter in UBR60 cells was completely inhibited by acute suppression of BRCA1 by RNA interference. These data reveal a role of BRCA1 in maintaining genome integrity by interplaying with p53 and genes that are involved in the spindle checkpoint and apoptosis.

Accumulating epidemiological studies have implicated a strong link between age associated metabolic diseases and cancer, though direct and irrefutable evidence is missing. In this review, we discuss the connection between Warburg effects and tumorigenesis, as well as adaptive responses to environment such as circadian rhythms on molecular pathways involved in metabolism. We also review the central role of the sirtuin family of proteins in physiological modulation of cellular processes and age-associated metabolic diseases. We also provide a macroscopic view of how the circadian rhythm affects metabolism and may be involved in cell metabolism reprogramming and cancer pathogenesis. The aberrations in metabolism and the circadian system may lead to age-associated diseases directly or through intermediates. These intermediates may be either mutated or reprogrammed, thus becoming responsible for chromatin modification and oncogene transcription. Integration of circadian rhythm and metabolic reprogramming in the holistic understanding of metabolic diseases and cancer may provide additional insights into human diseases.

Premise of the study: To elucidate the population genetics and phylogeography of Scrophularia incisa, microsatellite primers were developed. We also applied these microsatellite markers to its closely related species S. dentata and S. kiriloviana. Methods and Results: Using the compound microsatellite marker technique, 12 microsatellite primers were identified in S. incisa. The number of alleles ranged from 14 to 26 when assessed in 78 individuals from four populations. With high cross-species transferability, these primers also amplified in S. dentata and S. kiriloviana. Conclusions: These results indicate that these microsatellite markers are adequate for detecting and characterizing population genetic structure in the Chinese species of sect. Tomiophyllum at fine and range-wide geographical scales.

Protein phosphatase 1 (PP1) plays important roles in many different aspects of cellular activities including cell cycle control. One important function of PP1 is to activate the retinoblastoma protein pRB. Here we show that pRB is one of PP1's downstream targets during apoptosis. When HL-60 cells synchronized at the G1/S boundary were treated with pro-apoptotic cytosine arabinoside (araC), PP1alpha protein increased twofold and PP1 activity about 30% within 1 h. This was followed by pRB dephosphorylation, pRB cleavage by caspases, DNA fragmentation, the appearance of cells with

Our study examines the effect of apoptosis on prothymosin alpha, an abundant, nuclear protein intimately involved with proliferation of all mammalian cells. When HeLa cells were treated with actinomycin D, with etoposide, or with staurosporine following synchronization with hydroxyurea, they underwent apoptosis based on several specific criteria, including fragmentation of DNA and activation of specific caspases. Similarly treated NIH3T3 cells arrested and displayed no indicators of apoptosis. In HeLa, but not in NIH3T3 cells, prothymosin alpha levels declined precipitously and a truncated version of the protein was formed. The following observations implicate caspase activity: (1) The truncated polypeptide arose only in the treated HeLa cell cultures. (2) The appearance of the truncated polypeptide coincided with the activation of caspase 3 and the cleavage of poly(ADP-ribose) polymerase, a known caspase substrate. (3) Carbobenzoxy-DEVD-fluoromethylketone, a cell-permeable caspase 3 inhibitor, blocked cleavage and degradation of prothymosin alpha. (4) The same inhibitor, when added to mixed extracts of apoptotic and normal cells, prevented cleavage of intact prothymosin alpha. (5) Recombinant caspase 3 and, to a much lesser extent, caspase 7 truncated purified prothymosin alpha. (6) In HeLa cells, cleavage occurred at three overlapping caspase 3-like sites with the consensus sequence D-X-X-D and released 10 to 14 residues from the carboxyl terminus, including the core nuclear localization signal. Two immediate consequences of the cleavage were observed: truncated prothymosin alpha was no longer confined to the nucleus and it was deficient in phosphate. These data suggest that the disabling of prothymosin alpha is a significant event in apoptosis. J. Cell. Physiol. 182:256-268, 2000. Published 2000 Wiley-Liss, Inc.

Prothymosin alpha gene expression accompanies growth of all mammalian cells. The protein, which is abundant, exceedingly acidic, and localized to the nucleus, is further distinguished by the presence of clustered phosphorylated glutamic acid residues (Trumbore et al., 1997, J Biol Chem 272:26394-26404). These glutamyl phosphates are energy rich and unstable in vivo and in vitro (Wang et al., 1997, J Biol Chem 272:26405-26412). To understand the function of prothymosin alpha in greater detail, the turnover of its phosphates was examined in metabolically manipulated cells. Phosphate half-lives in growing, mock transfected, and vector-transfected COS cells were compared with the half-life in cells transfected with the prothymosin alpha gene to determine the fate of the predominantly ectopic phosphorylated protein. The values obtained--72-75 min in cells with normal levels of the protein, but 118 min in cells with surplus prothymosin alpha--led us to conclude that underutilized phosphates persist whereas functioning phosphates disperse. Cell-cycle-specific differences in the half-lives were observed in NIH3T3 cells: 72 min while cycling, 83 or 89 min during arrest in or progression through S phase, but 174 min during M-phase arrest. In the presence of actinomycin D, the value was about 145 min regardless of whether cells were quiescent or growing. In these experiments, reduced utilization of prothymosin alpha's glutamyl phosphates, signaled by an increase in their half-lives, accompanied the attenuation or abolition of transcription. Our data suggest that prothymosin alpha fuels an energy-requiring step in the production, processing, or export of RNA.

Objective To observe the influence of transcutaneous CO2 emission (TCE) at points on the meridian or beside the meridian after different stimulations like acupuncture De-qi, sham acupuncture, study the correlation between acupuncture De-qi and local energy metabolism through TCE.

SummaryDNA damage elicits a cellular signaling response that initiates cell cycle arrest and DNA repair. Here, we find that DNA damage triggers a critical block in glutamine metabolism, which is required for proper DNA damage responses. This block requires the mitochondrial SIRT4, which is induced by numerous genotoxic agents and represses the metabolism of glutamine into tricarboxylic acid cycle. SIRT4 loss leads to both increased glutamine-dependent proliferation and stress-induced genomic instability, resulting in tumorigenic phenotypes. Moreover, SIRT4 knockout mice spontaneously develop lung tumors. Our data uncover SIRT4 as an important component of the DNA damage response pathway that orchestrates a metabolic block in glutamine metabolism, cell cycle arrest, and tumor suppression.

The proposed relativistic hadronic cascade model has been used to analyse two-pion Bose-Einstein correlation in central collision of Si + Au at AGS energy. Theπ−π− correlation function calculated according to the space-time and four momentum distributions ofπ− at freeze-out agrees quite well with that of E802 experimental onse. We also have calculated the root mean squared length of theπ− source above and compared it with the conventional fitted radius ofπ− source. It is mentioned that due to the non-Gaussian features in position and momentum distributions of the source and the dependence of the location upon the momentum especially, to regard the conventional fitted radius as the spatial extent of the source is dangerous. Instead, it might be more reliable to predict the spatial extent of the source with the root mean squared length.

Patient-specific induced pluripotent stem cells (iPSCs) represent a potential source for developing novel drugand cell- therapies. Although increasing numbers of disease-specific iPSCs have been generated, there has been limited progress in iPSC-based drug screening/discovery for liver diseases, and the low gene targeting efficiency in human iPSCs warrants further improvement. Using iPSC lines from patients with alpha-1 antitrypsin (AAT) deficiency, for which there is currently no drug- or gene- therapy available, we established a platform to discover new drug candidates and to correct disease-causing mutation with a high efficiency. A high-throughput format screening assay based on our hepatic differentiation protocol was implemented to facilitate automated quantification of cellular AAT accumulation using a 96-well immunofluorescence reader. To expedite the eventual application of lead compounds to patients, we conducted drug screening utilizing our established library of clinical compounds, the Johns Hopkins Drug Library, with extensive safety profiles. Through a blind large-scale drug screening, five clinical drugs were identified to reduce AAT accumulation in diverse patient iPSC-derived hepatocyte-like cells. In addition, using the recently developed transcription activator-like effector nuclease (TALEN) technology, we achieved high gene targeting efficiency in AAT-deficiency patient iPSCs with 25–33% of the clones demonstrating simultaneous targeting at both diseased alleles. The hepatocyte-like cells derived from the gene-corrected iPSCs were functional without the mutant AAT accumulation. This highly efficient and cost-effective targeting technology will broadly benefit both basic and translational applications. Conclusions: Our results demonstrated the feasibility of effective large-scale drug screening using an iPSC-based disease model and highly robust gene targeting in human iPSCs; both of which are critical for translating the iPSC technology into novel therapies for untreatable diseases.

Smilax Linnaeus (1753: 1028) is the only genus of Smilacaceae, comprising ca. 210 species, which are lianas, shrubs, and herbs (Cameron & Fu, 2006; Qi et al. 2013a). The genus shows a near-cosmopolitan distribution (Qi et al. 2013a). In the large genus, Smilax sect. Heterosmilax (Kunth 1850: 270) Judd (1998: 158), formerly treated as an independent genus Heterosmilax Kunth in Smilacaceae, has 12 species and is distributed only in Asian and Malesian regions (Judd 1998, Qi et al. 2013b). All species of the section have slender stem without pricks, cordate leaves, and tendrils in bronze color. Especially, flowers with connate perigone are one of the most important characteristics to distinguish the section from the other sections (Koyama 1984, Qi et al. 2013a).

Based on principal component analysis (PCA), this paper proposed a new method for studying the current situation of the basic public service in HeBei province. The authors evaluated the basic public service development level of the province by establishing a comprehensive evaluation index system. The system employed the combination of PCA and factor analysis. Numerical analysis was carried out using the quantized data based on the system. Analysis results showed that the level of the basic public services is below the national average level, especially in the medical services, health and basic education services.

SummaryMembers of sirtuin family regulate multiple critical biological processes, yet their role in carcinogenesis remains controversial. To investigate the physiological functions of SIRT2 in development and tumorigenesis, we disrupted Sirt2 in mice. We demonstrated that SIRT2 regulates the anaphase-promoting complex/cyclosome activity through deacetylation of its coactivators, APCCDH1 and CDC20. SIRT2 deficiency caused increased levels of mitotic regulators, including Aurora-A and -B that direct centrosome amplification, aneuploidy, and mitotic cell death. Sirt2-deficient mice develop gender-specific tumorigenesis, with females primarily developing mammary tumors, and males developing more hepatocellular carcinoma (HCC). Human breast cancers and HCC samples exhibited reduced SIRT2 levels compared with normal tissues. These data demonstrate that SIRT2 is a tumor suppressor through its role in regulating mitosis and genome integrity.

BRCA1, a well-known tumor suppressor with multiple interacting partners, is predicted to have diverse biological functions. However, so far its only well-established role is in the repair of damaged DNA and cell cycle regulation. In this regard, the etiopathological study of low-penetrant variants of BRCA1 provides an opportunity to uncover its other physiologically important functions. Using this rationale, we studied the R1699Q variant of BRCA1, a potentially moderate-risk variant, and found that it does not impair DNA damage repair but abrogates the repression of microRNA-155 (miR-155), a bona fide oncomir. Mechanistically, we found that BRCA1 epigenetically represses miR-155 expression via its association with HDAC2, which deacetylates histones H2A and H3 on the miR-155 promoter. We show that overexpression of miR-155 accelerates whereas the knockdown of miR-155 attenuates the growth of tumor cell lines in vivo. Our findings demonstrate a new mode of tumor suppression by BRCA1 and suggest that miR-155 is a potential therapeutic target for BRCA1-deficient tumors.

Insulin resistance is a major risk factor for type 2 diabetes mellitus. The protein encoded by the sirtuin 1 (Sirt1) gene, which is a mouse homolog of yeast Sir2, is implicated in the regulation of glucose metabolism and insulin sensitivity; however, the underlying mechanism remains elusive. Here, using mice with a liver-specific null mutation of Sirt1, we have identified a signaling pathway involving Sirt1, Rictor (a component of mTOR complex 2 [mTorc2]), Akt, and Foxo1 that regulates gluconeogenesis. We found that Sirt1 positively regulates transcription of the gene encoding Rictor, triggering a cascade of phosphorylation of Akt at S473 and Foxo1 at S253 and resulting in decreased transcription of the gluconeogenic genes glucose-6-phosphatase (G6pase) and phosphoenolpyruvate carboxykinase (Pepck). Liver-specific Sirt1 deficiency caused hepatic glucose overproduction, chronic hyperglycemia, and increased ROS production. This oxidative stress disrupted mTorc2 and impaired mTorc2/Akt signaling in other insulin-sensitive organs, leading to insulin resistance that could be largely reversed with antioxidant treatment. These data delineate a pathway through which Sirt1 maintains insulin sensitivity and suggest that treatment with antioxidants might provide protection against progressive insulin resistance in older human populations.

SIRT1 is a founding member of a sirtuin family of 7 proteins and histone deacetylases. It is involved in cellular resistance to stress, metabolism, differentiation, aging, and tumor suppression. SIRT1−/− mice demonstrate embryonic and postnatal development defects. We examined hematopoietic and endothelial cell differentiation of SIRT1−/− mouse embryonic stem cells (ESCs) in vitro, and hematopoietic progenitors in SIRT1+/++/−, and −/− mice. SIRT1−/− ESCs formed fewer mature blast cell colonies. Replated SIRT1−/− blast colony-forming cells demonstrated defective hematopoietic potential. Endothelial cell production was unaltered, but there were defects in formation of a primitive vascular network from SIRT1−/−-derived embryoid bodies. Development of primitive and definitive progenitors derived from SIRT1−/− ESCs were also delayed and/or defective. Differentiation delay/defects were associated with delayed capacity to switch off Oct4, Nanog and Fgf5 expression, decreased β-H1 globin, β-major globin, and Scl gene expression, and reduced activation of Erk1/2. Ectopic expression of SIRT1 rescued SIRT1−/− ESC differentiation deficiencies. SIRT1−/− yolk sacs manifested fewer primitive erythroid precursors. SIRT1−/− and SIRT1+/− adult marrow had decreased numbers and cycling of hematopoietic progenitors, effects more apparent at 5%, than at 20%, oxygen tension, and these progenitors survived less well in vitro under conditions of delayed growth factor addition. This suggests a role for SIRT1 in ESC differentiation and mouse hematopoiesis.

Glucose homeostasis in mammals is mainly regulated by insulin signaling. It was previously shown that SIRT6 mutant mice die before 4 weeks of age, displaying profound abnormalities, including low insulin, hypoglycemia, and premature aging. To investigate mechanisms underlying the pleiotropic phenotypes associated with SIRT6 deficiency, we generated mice carrying targeted disruption of SIRT6. We found that 60% of SIRT6(-/-) animals had very low levels of blood glucose and died shortly after weaning. The remaining animals, which have relatively higher concentrations of glucose, survived the early post-weaning lethality, but most died within one year of age. Significantly, feeding the mice with glucose-containing water increased blood glucose and rescued 83% of mutant mice, suggesting that the hypoglycemia is a major cause for the lethality. We showed that SIRT6 deficiency results in more abundant membrane association of glucose transporters 1 and 4, which enhances glucose uptake. We further demonstrated that SIRT6 negatively regulates AKT phosphorylation at Ser-473 and Thr-308 through inhibition of multiple upstream molecules, including insulin receptor, IRS1, and IRS2. The absence of SIRT6, consequently, enhances insulin signaling and activation of AKT, leading to hypoglycemia. These data uncover an essential role of SIRT6 in modulating glucose metabolism through mediating insulin sensitivity.

Clinical application of retinoic acids (RAs) and demethylation agents has proven to be effective in treating certain myeloid leukemia patients. However, the target genes that mediate these antileukemia activities are still poorly understood. In this study, we identified olfactomedin 4 (OLFM4), a myeloid-lineage–specific gene from the olfactomedin family, as a novel target gene for RAs and the demethylation agent, 5-aza-2′-deoxycytidine. We demonstrated that the retinoic acid receptor alpha/retinoic X receptor alpha heterodimer binds to a retinoic acid response-element (DR5) site in the OLFM4 promoter and mediates all-trans-retinoic acid (ATRA)–induced transactivation of the OLFM4 gene. OLFM4 overexpression in HL-60 cells led to growth inhibition, differentiation, and apoptosis, and potentiated ATRA induction of these effects. Conversely, down-regulation of endogenous OLFM4 in acute myeloid leukemia-193 cells compromised ATRA-induced growth inhibition, differentiation, and apoptosis. Overexpression of OLFM4 in HL-60 cells inhibited constitutive and ATRA-induced phosphorylation of the eukaryote initiation factor 4E-binding protein 1 (4E-BP1), whereas down-regulation of OLFM4 protein in acute myeloid leukemia-193 cells increased 4E-BP1 phosphorylation, suggesting that OLFM4 is a potent upstream inhibitor of 4E-BP1 phosphorylation/deactivation. Thus, our study demonstrates that OLFM4 plays an important role in myeloid leukemia cellular functions and induction of OLFM4-mediated effects may contribute to the therapeutic value of ATRA.

Olfactomedin 4 (OLFM4) is a glycoprotein that has been found to be up-regulated in inflammatory bowel diseases and Helicobacter pylori infected patients. However, its role in biological processes such as inflammation or other immune response is not known. In this study, we generated OLFM4 KO mice to investigate potential role(s) of OLFM4 in gastric mucosal responses to H. pylori infection. H. pylori colonization in the gastric mucosa of OLFM4 KO mice was significantly lower compared with WT littermates. The reduced bacterial load was associated with enhanced infiltration of inflammatory cells in gastric mucosa. Production and expression of proinflammatory cytokines/chemokines such as IL-1β, IL-5, IL-12 p70, and MIP-1α was increased in OLFM4 KO mice compared with infected controls. Furthermore, we found that OLFM4 is a target gene of NF--κB pathway and has a negative feedback effect on NF-κB activation induced by H. pylori infection through a direct association with nucleotide oligomerization domain-1 (NOD1) and -2 (NOD2). Together these observations indicate that OLFM4 exerts considerable influence on the host defense against H. pylori infection acting through NOD1 and NOD2 mediated NF-κB activation and subsequent cytokines and chemokines production, which in turn inhibit host immune response and contribute to persistence of H. pylori colonization.

While a Hall-Petch-type dependence is known for deformation twinning (DT) in Cu and other metals of conventional grain sizes $(Dg1\text{ }\ensuremath{\mu}\text{m})$, with $D$ decreasing into the nanocrystalline regime, the propensity for DT turns around to increase, exhibiting an inverse grain size dependence. This trend is inversed again at still smaller grain sizes, returning to the behavior of increased difficulty in DT with $D$ going further down. This double-inverse behavior with respect to the normal Hall-Petch $D$ dependence is demonstrated here for nanocrystalline Cu films, deformed in tension at room temperature and slow strain rates. The nonmonotonic $D$ dependence of DT is explained by modeling the competing grain size effects on the emission of the first partial dislocation and the plane-to-plane promotion of partial dislocation slip afterwards.

By using Hoechst 33342, rabbit anti calmodulin antibody, FITC-labeled goat anti rabbit IgG and SR101 (sulfo rhodamine 101) simultaneously to stain individual normal and transformed cells, the microspectrophotometric analysis demonstrated that 3 markers which represented the nucleus, calmodulin and total protein respectively, could be recognized in individualj cells without interference. The phase of the cell cycle was determined by DNA content (Hoechst 33342). We found that in transformed cells (NIH3T3) tsRSV-LA90, cultured at 33°C and transformed C3H10T1/2 cells), the ratio of calmodulin to total protein (based on the phases of cell cycle)was higher than that in normal cells (NIH3T3 tsRSV−LA90 cells, cultured at 39°C and C3H10T1/2 cells)in every cell cycle phase. This ratio increased obviously only from G1 to S phase in either normal or transformed cells. The results showed that calmodulin really increased during the transformation, and its increase was specific. In the meantime when cells proceeded from G1 to S, the intracellular calmodulin content also increased specifically.

To investigate conduction of acupuncture effect and the influence of mechanical pressing meridians on it.In twenty healthy volunteers (8 males and 12 females, aged 29 years on average), by using a Frewil Q. F. apparatus, transcutaneous CO2 emission (TCE) was measured before, during and after acupuncture at Neiguan (PC 6), respectively. The locations being measured were Quze (PC 3) and four points including two non-acupoints along the Pricardium Meridian (Point A and B) and two non-acupoints apart from the Pericardium Meridian (Point C and D). Point A was located at the lower one fourth of the line connecting Tianquan (PC 2) and Quze (PC 3), and Point B located at the middle of the line connecting Ximen (PC 4) and Quze (PC 3). Point C was located 2 cm lateral on the ulnar side to Point A, and Point D located 2 cm lateral on the ulnar side to Point B. The above results were compared with those obtained after mechanical pressure of 1 kg (area 1 cm X 2 cm) applied on Ximen (PC 4) and Daling (PC 7), respectively, to observe their effect on TCE.During and after acupuncture at Neiguan (PC 6) without mechanical pressure, TCE at Quze (PC 3), point A and B increased significantly (P0.05), but not at point C and D (P0.05). While applying mechanical pressure at Ximen (PC 4), no significant increase was found on the three points during and after acupuncture. When applying pressure at Daling (PC 7), significant increases of TCE were also found at Quze (PC 3) and point A during acupuncture (P0.05), and a significant increase of TCE was found at point B after acupuncture with pressure keeping (P0.05).Acupuncture can enhance energy metabolism along meridians, and this effect can be blocked by mechanical pressure.