Irradiation resistance mechanism of the CoCrFeMnNi equiatomic high-entropy alloy.

When face-centered cubic (FCC) metals and alloys with low stacking fault energy (SFE) are irradiated by high-energy particles or deformed at high speed, stacking fault tetrahedra (SFTs), which are a type of vacancy cluster defect, are often formed. Therefore, SFTs were expected to form in the CoCrFeMnNi equiatomic high-entropy alloy (HEA). However, no SFT was observed in the CoCrFeMnNi HEA with high-speed plastic deformation even after annealing at 873 K. To elucidate this mechanism, the binding energy of vacancy clusters in the CoCrFeMnNi HEA was calculated based on first principles. The binding energy of the di-vacancy cluster was positive (average of 0.25 eV), while that of the tri-vacancy cluster was negative (average of - 0.44 eV), suggesting that the possibility of formation of a tri-vacancy cluster was low. The inability to form a cluster containing three vacancies is attributed to the excellent irradiation resistance of the CoCrFeMnNi HEA. However, if an extra vacancy is added to a tri-vacancy cluster (with negative binding energy), the binding energy of the subsequent tetra-vacancy cluster may become positive. This suggests that it is possible to form vacancy clusters in the CoCrFeMnNi HEA when high-energy ion or neutron irradiation causes cascade damage.

[Prevalence and risk factors of hypertension combined with diabetes in middle and elder population in Nan'an district of Chongqing].

Objective: To analyze the prevalence and risk factors of hypertension combined with diabetes in the middle to elder population in the Nan'an district of Chongqing, and to provide evidence for formulating relevant prevention and control strategies. Methods: Middle or elder adults were enrolled by a Stratified multistage cluster sampling method. Questionnaire survey and the related measurements were conducted. The epidemiology of hypertension combined with diabetes was analyzed descriptively, and the risk or protective factors were analyzed by logistic regression method. Results: A total of 24 792 people were surveyed, with 1 547 patients identified as having hypertension combined with diabetes. The overall prevalence rate appeared as 6.2%, of which 6.0% in males and 6.4% in females, respectively. The prevalence of hypertension combined with diabetes in the general population was increasing with age (χ(2)=343.766, P<0.001). Factors as age, education, smoking, marital status, exercise, BMI, triglyceride, total cholesterol, high density lipoprotein cholesterol and low density lipoprotein cholesterol were related to the prevalence of hypertension and diabetes. High density lipoprotein cholesterol appeared as a protective factor for hypertension combined with diabetes (OR=0.817, 95%CI: 0.715-0.934). Age, education, low density lipoprotein cholesterol and lack of exercise all appeared as risk factors for hypertension combined with diabetes (P<0.05), respectively. Conclusions: The prevalence rate of hypertension combined with diabetes in the middle or elder adults in Nan'an of Chongqing seemed high. Attention should be paid to the health status of people being elderly, overweight or obese, low cultural level, smoking, triglyceride abnormality, total cholesterol abnormality and high low density lipoprotein cholesterol, so as to reduce the risk on hypertension combined with diabetes.

Evidence for Secondary Flux Rope Generated by the Electron Kelvin-Helmholtz Instability in a Magnetic Reconnection Diffusion Region.

Secondary flux ropes are suggested to play important roles in energy dissipation and particle acceleration during magnetic reconnection. However, their generation mechanism is not fully understood. In this Letter, we present the first direct evidence that a secondary flux rope was generated due to the evolution of an electron vortex, which was driven by the electron Kelvin-Helmholtz instability in an ion diffusion region as observed by the Magnetospheric Multiscale mission. The subion scale (less than the ion inertial length) flux rope was embedded within the electron vortex, which contained a secondary electron diffusion region at the trailing edge of the flux rope. We propose that intense electron shear flow produced by reconnection generated the electron Kelvin-Helmholtz vortex, which induced a secondary reconnection in the exhaust of the primary X line and then led to the formation of the flux rope. This result strongly suggests that secondary electron Kelvin-Helmholtz instability is important for reconnection dynamics.

The expression of inhibitor of growth 4 is reduced in cervical cancer tissues.

OBJECTIVE: Inhibitor of growth 4 (ING4) is a candidate tumor suppressor which plays an important role in multiple processes including DNA repair, apoptosis, cell cycle control, tumor metastasis and angiogenesis. However, clinical data about the role of ING4 in the development and progression of cervical cancer are still limited. This study aimed to examine ING4 expression in cervical cancer and analyze its correlation with the progression of the malignancy. PATIENTS AND METHODS: RT-PCR, Western blot and immunohistochemistry analysis were performed to determine ING4 expression in 18 clinical specimens from cervical cancer patients. The correlation of ING4 expression with the clinical-pathological features of the patients was analyzed. Moreover, the correlation between ING4 and HPV E6/E7 transcription level in SiHa cells was analyzed. RESULTS: ING4 expression was decreased significantly at mRNA and protein levels in the tissues of cervical cancer compared with paracarcinoma tissues. Analysis of the subcellular localization of ING4 showed that ING4 expression was decreased in the nucleus of cervical cancer tissues. Ectopic expression of ING4 reduced the proliferation of SiHa cells, accompanied by decreased HPV E6/E7 transcription. CONCLUSIONS: ING4 expression is decreased in human cervical cancer tissues. Reconstitution of ING4 expression in cervical cancer cells is correlated with decreased HPV E6/E7 transcription. These data suggest that ING4 expression has diagnostic and prognostic significance for cervical cancer.

Exogenous expression of IL-1Ra and TGF-ß1 promotes in vivo repair in experimental rabbit osteoarthritis.

OBJECTIVES: Potential gene therapy application of single and co-expression of interleukin 1 (IL-1) receptor antagonist (IL-1Ra) and transforming growth factor-ß1 (TGF-ß1) to alter disease progression was investigated in an in-vivo rabbit model of osteoarthritis (OA). METHOD: Sixteen young adult rabbits were randomly and equally divided into four groups: blank control group, IL-1Ra transfection group, TGF-ß1 transfection group, and IL-1Ra/TGF-ß1 double transfection group. Histological examinations were performed to monitor disease progression after haematoxylin and eosin (H&E) staining of articular cartilage. Immunohistochemistry was used to detect IL-1Ra and TGF-ß1 in synovial membrane tissues. Exogenous IL-1Ra and TGF-ß1 content was assessed in joint lavage fluid using an enzyme-linked immunosorbent assay (ELISA). RESULTS: ELISA measurements from the joint lavage fluid showed high expressions of IL-1Ra and TGF-ß1 in the single and double transfection groups. Remarkably, concomitant reductions in IL-1ß and tumour necrosis factor alpha (TNF-α) levels were observed in these single and double transfection groups. Radioimmunoassay (RIA)-based detection showed that IL-1ß and TNF-α levels in the gene transfection groups were significantly lower compared to the blank control group, in parallel experiments. Importantly, injection of IL-1Ra and TGF-ß1 expressing cartilage cells into joints led to a significant inhibition of cartilage matrix degradation. Finally, IL-1Ra and TGF-ß1 expression in tissues correlated with disease reversal in the experimental group, with improved tissue architecture and collagen deposition. CONCLUSIONS: Our results reveal that both single- and double-gene transfection of IL-1Ra and TGF-ß1 promote extensive repair of damaged cartilage, and double transfections showed better recovery than single transfections, suggesting that co-expression of IL-1Ra and TGF-ß1 inhibits degeneration and improves repair of articular cartilage in OA.

Creep mechanisms of a new Ni-Co-base disc superalloy at an intermediate temperature.

The microstructures of a new Ni-Co-base disc superalloy, TMW-4M3, before and after the creep test at 725 °C/630 MPa have been systematically investigated by transmission electron microscopy (TEM). The crept microstructures were marked as three different deformation stages (I, II and III) corresponding to the gradually increased strain. At stage I, stacking fault (SF) shearing was the main deformation mechanism. The SF was extrinsic and lay on {111} plane. However, deformation microtwinning became the dominant mode at stage II and III. The average spacing of deformation twins decreased from 109 ± 15 nm at stage II to 76 ± 12 nm at stage III, whereas the twin thickness did not change significantly. The influence of stacking fault energy (SFE) of γ matrix on the deformation mechanism is discussed. It is suggested that lower SFE in TMW-4M3 is partly responsible for the enhanced creep resistance.

Osteogenic growth peptide accelerates bone healing during distraction osteogenesis in rabbit tibia.

Distraction osteogenesis is a valuable treatment method that allows limb lengthening or reconstruction of large bone defects. However, its major disadvantage is the long period required for the consolidation of a distraction callus. Osteogenic growth peptide (OGP) stimulates endochondral bone formation in fracture callus, but its capacity to promote regenerate ossification during distraction osteogenesis has not been evaluated. This study investigated whether intravenously administered OGP accelerated bone healing during distraction osteogenesis in 36 male New Zealand White rabbits, randomized into two groups. The treatment group received OGP (200 ng/kg body weight) in phosphate-buffered saline (PBS), intravenously, each day; the control group received PBS alone. A 15-mm lengthening of the right lower leg was performed using the method of Ilizarov. Evidence from biomechanical, histological and radiographic evaluations demonstrated that systemic OGP treatment promoted optimal new bone formation during distraction osteogenesis in this rabbit model.

Upregulation of survivin during immortalization of nontransformed human fibroblasts transduced with telomerase reverse transcriptase.

These investigations demonstrate that expression of the inhibitor of apoptosis family member, survivin, is dramatically increased during immortalization of nontransformed human fibroblasts that were transduced with telomerase reverse transcriptase (hTERT). Expression of survivin in immortalized fibroblasts peaked during G(2)/M phase of the cell cycle. However, the upregulation of survivin was dissociated from the rate of proliferation and proportion of G(2)/M cells. Depletion of survivin from immortal fibroblasts increased sensitivity to stress-induced apoptosis and resulted in an accumulation of cells with 4N DNA content. Conversely, overexpression of survivin in mortal fibroblasts conferred resistance to apoptosis. In contrast, very low levels of survivin in proliferating parental fibroblasts had no bearing on sensitivity to apoptosis. The upregulation of survivin did not appear to be a direct consequence of hTERT transduction. However, repression of hTERT resulted in the rapid downregulation of survivin in telomerase-immortalized fibroblasts and tumor cell lines, but not in cells immortalized via an Alternative Lengthening of Telomeres mechanism. These results have important therapeutic implications, as telomerase and survivin are both broadly expressed in human cancers. Selection during the immortalization process for cells expressing high levels of survivin may account for the abundance of survivin in diverse tumor types.

Efficacy of c-erbB-2 antisense oligonucleotide transfection on uterine endometrial cancer HEC-1A cell lines.

OBJECTIVE: Antigene therapy targeting only one oncogene has made much progress although it still has some limitations. To explore the potential for antigene therapy in uterine endometrial cancer, we examined the in vitro inhibitory effects of liposmal anti-sense phosphorothioate oligonucleotides targeting c-erbB-2 in the human uterine endometrial cancer HEC-1A cell line. METHODS: 1) To detect c-erbB-2 protein expression on HEC-1A cell membranes by immunohisto- chemistry. 2) To assay cellular growth inhibition by MTT after transfecting 0.1-0.6 microM ASODN. 3) To observe cellular and ultra-structural changes under transmission electron microscope and to assay the cellular apoptotic rate by flow cytometry and c-erbB-2 mRNA, and protein expression by RT-PCR and Western blot after transfecting 0.3 microM ASODN. RESULTS: 1) c-erbB-2 protein expression was positive on HEC-1A cell membranes. 2) With the increase of the transfecting ASODN concentration from 0.1-0.6 microM, HEC-1A cellular growth inhibition was also enhanced. The results of MTT showed that when the transfecting concentration of ASODN was 0.3 microM, the HEC-1A cellular growth inhibition rate was 50% while when the transfecting concentration of ASODN was 0.6 microM, the HEC-1A cell growth inhibition rate was 75%. 3) When the concentration of transfecting ASODNs was 0.3 microM, there were obvious vacuolar degenerations in the plasma of HEC-1A cells, disappearance of organelle and nuclear structure and obvious shrinkage of nuclei under transmission electron microscope. The cellular apoptotic rate was 62.80%, while c-erbB-2 mRNA and protein expression were 47.18% and 33.60%, respectively, compared with those of the normal control cells. CONCLUSION: Transfecting c-erbB-2 ASODNs can obviously suppress the mRNA and protein expression in HEC-1A cells, cause cellular apoptosis and inhibit cell growth. It may be a more useful gene therapy for endometrial cancer.

Identification of candidate alternative lengthening of telomeres genes by methionine restriction and RNA interference.

Telomerase-negative cancer cells can maintain their telomeres by a recombination-mediated alternative lengthening of telomeres (ALT) process. We reported previously that sequestration of MRE11/RAD50/NBS1 complexes represses ALT-mediated telomere length maintenance, and suppresses formation of ALT-associated promyelocytic leukemia (PML) bodies (APBs). APBs are PML bodies containing telomeric DNA and telomere-binding proteins, and are observed only in a small fraction of cells within asynchronously dividing ALT-positive cell populations. Here, we report that methionine restriction caused a reversible arrest in G0/G1 phase of the cell cycle and reversible induction of APB formation in most cells within an ALT-positive population. We combined methionine restriction with RNA interference to test whether the following proteins are required for APB formation: PML body-associated proteins, PML and Sp100; telomere-associated proteins, TRF1, TRF2, TIN2 and RAP1; and DNA repair proteins, MRE11, RAD50, NBS1 and 53BP1. APB formation was not decreased by depletion of Sp100 (as reported previously) or of 53BP1, although 53BP1 partially colocalizes with APBs. Depletion of the other proteins suppressed APB formation. Because of the close linkage between ALT-mediated telomere maintenance and ability to form APBs, the eight proteins identified by this screen as being required for APB formation are also likely to be required for the ALT mechanism.

Expression of interleukin-17 in ischemic brain tissue.

Ischemic brain injury is acute local inflammation, leading to accumulation of pro-inflammatory cytokines. Cytokines influence the recruitment of leucocytes and play a key role in the inflammatory injury processes. Recently, a number of studies have demonstrated a close relationship between brain ischemia and cytokines. Interleukin-17 (IL-17) is a newly identified T-cell-specific cytokine. In this study, we evaluated the source and the action of IL-17 over the course of cerebral ischemia in rats (Sprague-Dawley) and humans. The levels of IL-17 in the ischemic hemisphere of the human brain, which was removed at necropsy, were assayed immunohistochemically. In rats, permanent middle cerebral artery occlusion (pMCAO) was obtained by inserting nylon monofilament into the right external carotid artery, occluding the right middle cerebral artery. The expression of IL-17 mRNA in rat was assayed using oligoprobe in situ hybridization. IL-17 production by neuroglial cells was assayed by double-staining using antibody glial fibrillary acidic protein (GFAP) and antibody IL-17. Levels of IL-17 were elevated in the ischemic hemispheres of human brain compared with the opposite normal hemispheres and peaked at days 3-5 after brain ischemia. The IL-17-positive cells were found in the ischemic lesion region. IL-17 mRNA was also elevated in ischemic hemispheres of pMCAO-operated rats, which were slightly elevated after 1 h and peaked at 6 days. IL-17 and GFAP double-stained were extensive in rat ischemic hemisphere. The ischemia-induced IL-17 expression in human brain reported here for the first time was very similar to that in rat model except that the peak was slightly earlier. We found for the first time that IL-17 was involved in an intense inflammatory reaction of brain ischemic injury in human. In pMCAO-operated rats, our findings suggest that IL-17 is produced by the neuroglial cells in the brain region undergoing ischemic insult. We suggest that in additional to T cells the neuroglial cell may be another cellular origin of IL-17 in later progression of brain ischemia.

Insulin binding monitored by fluorescence correlation spectroscopy.

AIM/HYPOTHESIS: The characteristics of insulin binding to its receptors have been extensively studied by the radioligand binding assay. We used fluorescence correlation spectroscopy to determine the distribution of diffusion times and further novel data on the kinetics of insulin's binding to its receptor. METHODS: Cultured human renal tubular cells (HRTC) were incubated with tetramethyl rhodamine labelled insulin (Rh-Ins) for 60 min. Fluorescence intensity fluctuations and autocorrelation functions for Rh-Ins, free in the incubation medium and bound to the cell membrane, were studied at single-molecule detection sensitivity in a 0.2 fL confocal volume. RESULTS: Measurements at the cell membrane revealed Rh-Ins binding with at least two diffusion components (diffusion times tauD1 = 0.8 ms, tauD2 = 20 ms) and corresponding weight fractions of y1 = 0.43 and y2 = 0.42. Specificity of the binding was shown by the dislocation of bound Rh-Ins when excess unlabelled insulin was added. Scatchard analysis showed a nonlinear plot, revealing two binding processes with different affinities (Kass approximately 2 x 10(10) M(-1) and approximately 1 x 10(9) M(-1), respectively). CONCLUSION/INTERPRETATION: The fluorescence correlation spectroscopy results show two classes of binding sites with different affinities for insulin, or interactions between receptor sites consistent with negative cooperativity. This conclusion is in agreement with studies of insulin binding using radioligand binding assays. Because of its high sensitivity (single molecule detection), FCS, provides additional data allowing a more precise evaluation of the kinetics of ligand-receptor interactions at low expression levels in living cells.

Detection and differentiation of CAV-1 and CAV-2 by polymerase chain reaction.

Canine adenovirus type 1 (CAV-1) and type 2 (CAV-2) can be categorized in the laboratory by haemagglutination and neutralization tests, but they are difficult to differentiate from each other in specimens, especially when infection occurs in the digestive tract. The object of this study was to develop a simple method of detecting and differentiating them. One pair of common primers was designed and synthesized according to the sequences of the E3 and flanking regions and a polymerase chain reaction (PCR) assay was established using these two primers to amplify the virus-specific DNA fragment from clinical specimens as well as from cell cultures. After elecctrophoresis, under the same amplification conditions, 508 bp and 1030 bp PCR products were observed for CAV-1 and CAV-2, respectively. These were further shown to be adenovirus specific by dot hybridization and sequencing. As only one pair of primers was involved in the PCR procedure, it was faster and easier to perform than any of the other assays used for detecting canine adenovirus, making it applicable in the rapid confirmation of diagnosis and differentiation of the two types of canine adenoviruses.

Pharmacokinetics of nortriptyline and its 10-hydroxy metabolite in Chinese subjects of different CYP2D6 genotypes.

OBJECTIVES: To study the impact of the CYP2D6*10 allele on the disposition of nortriptyline in Chinese subjects. METHODS: A single dose of 25 mg nortriptyline was given orally to 15 healthy Chinese volunteers who were classified as extensive metabolizers after phenotyping with debrisoquin (INN, debrisoquine) and who were genotyped by allele-specific polymerase chain reaction. Five subjects were homozygous for CYP2D6*1, 5 subjects were homozygous for CYP2D6*10, and 5 subjects were heterozygous for these 2 alleles. Plasma concentrations of nortriptyline and its main metabolite 10-hydroxynortriptyline were measured by liquid chromatography-mass spectrometry, and the pharmacokinetics were studied during 168 hours after the dose. RESULTS: Subjects who were homozygous for CYP2D6*10 had significantly higher total areas under the plasma concentration-time curve (AUC), lower apparent oral clearances, and longer mean plasma half-life of nortriptyline than subjects in the CYP2D6*1/*1 and the heterozygous groups. For 10-hydroxynortriptyline, the AUC was lower and the plasma half-life was longer in subjects who were homozygous for CYP2D6*10 than in subjects in the other 2 groups. CONCLUSION: The CYP2D6*10 allele in Chinese subjects was associated with significantly higher plasma levels of nortriptyline compared with the CYP2D6*1 allele because of an impaired metabolism of nortriptyline to 10-hydroxynortriptyline, particularly in the subjects with the CYP2D6*10/*10 genotype. The results suggest that genotyping of CYP2D6 may be a useful tool in predicting the pharmacokinetics of nortriptyline.

[Effect of liuwei dihuang granules, pills and decoction on contents of cAMP, E2, T, Zn and Cu in plasma of renal yin deficient patients].

Experiments showed that in patients of renal yin deficiency who took orally Liuwei Dihuang granules for 4 weeks, the contents of cAMP, E2, Zn and Cu were found mankedly lower, but the contents of T mankedly higher than before using the drugs. This indicates that the granules are equally effective as pills and decoction.