Search

It has been proposed that thioredoxin reductase (TR) is a mediator that allows non-small cell lung cancer (NSCLC) to develop resistance to irradiation; however, little is known regarding the detailed mechanisms of action. Thus, ethaselen {1, 2-[bis (1,2-benzisoselenazolone-3 (2H)-ketone)] ethane, BBSKE}, a novel organoselenium TR inhibitor, is currently being investigated in a phase I clinical trial in China. However, its radiosensitizing effect remains unexplored. In this study, we found that the activity of TR increased dramatically in both A549 and H1299 cells after radiation, and moreover, could be inhibited by pretreatment with BBSKE (5 μmol/l). As a TR inhibitor, BBSKE enhanced the efficacy of radiation therapy both in vivo and in vitro without observable toxicity. BBSKE was found to suppress irradiation-induced NF-κB activation dramatically when using A549 cells stably transfected with NF-κB luciferase reporter. These results show the critical role of TR in the radioresistance of NSCLC and suggest that BBSKE is a potentially promising agent for the treatment of patients with NSCLC clinically.

We evaluated the combination treatment of ethaselen (BBSKE) as a thioredoxin reductase (TrxR) inhibitor plus cisplatin (CDDP) on the human colon adenocarcinoma cell line LoVo. Therapeutic effects ranging from nearly additive to clearly synergistic demonstrated an effective combination, i.e., the cytostatic dose of CDDP could be reduced without a loss in efficacy. To further investigate the cellular response mechanisms of these favorable outcomes, we analyzed the cell-cycle profiles, mRNA expression patterns, and protein levels of several key genes after incubation with BBSKE or CDDP separately and in combination. In appropriate conditions, CDDP induced arrest at the G2/M phase accompanied by the enhanced inhibitory phosphorylation of Cdk1 and the elevated protein expression of cyclin B1. BBSKE downregulated expression of cyclin D1 by increasing mRNA and protein levels of p21, and thus induced G1 phase arrest. BBSKE returned Cdk1 to an activated state, and reduced the protein level of cyclin B1 after incubation in combination with CDDP, which was consistent with the reduction in the percentage of cells in G2/M identified by flow cytometry. By regulating the G1 phase and reversing CDDP-induced G2/M phase arrest, BBSKE increases drug sensitivity of LoVo cells toward CDDP, and probably provides a meaningful anticancer strategy for further clinical studies.

A facile method based on HPLC/(-)ESI-MS(n) is established for the analysis of seven isomers of resveratrol dimers and three of their analogues in Xinjiang wine grapes. The structures of these compounds are positively or tentatively determined. Among them, three are tentatively identified as new compounds. MS(n) experiments on the [M-H](-) ions provide abundant structural information, especially regarding the relative abundance of the key product ions, m/z 333 and 369 (385 in compound 3), which can be utilised to distinguish whether or not the compound identified contains the scaffold of the isomer of a resveratrol dimer. The relative abundance of key product ions remains unchanged as collision energy varies from 0.60 to 0.95V. All the trans-, and cis-isomers could be identified by HPLC/DAD-UV spectra. The UV spectra of compounds 2 and 9 tentatively show cis and trans- configurations, respectively., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Aim: The aim of this study was to evaluate the effectiveness of low-volume zero-balanced ultrafiltration during cardiopulmonary bypass in heart valve replacement surgery., Methods: This was a randomized, double-blind, controlled study carried out in the operating room. Forty patients of ASA grade II-III, elected to undergo heart valve replacement surgery, were enrolled. All patients were randomly assigned to either a low-volume (35 mL/kg) zero-balanced ultrafiltration group (N.=20) or to a control group (N.=20). Blood propofol concentrations and entropy index were measured using cardiopulmonary bypass. Concentrations of plasma tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), interleukin-10 (IL-10), and cardiac troponin I were measured before and after the end of cardiopulmonary bypass and corrected according to hematocrit., Results: Blood levels of cardiac troponin I, TNF-alpha, IL-6, and IL-10 after surgery were all significantly lower in the ultrafiltration group than in the control group (P<0.05) after the end of bypass. Blood propofol concentrations decreased significantly in both groups during cardiopulmonary bypass and remained significantly lower in the ultrafiltration group than the control group. However, there was no significant difference between the two groups in the entropy index (P=0.5583)., Conclusion: Low-volume zero-balanced ultrafiltration performed during cardiopulmonary bypass surgery significantly decreased post-bypass levels of the cytokines, TNF-alpha, IL-6, IL-10, and postoperative cardiac troponin I. Blood propofol concentration was also decreased; however, the depth of anesthesia was not affected significantly.

Objective: To analyze the correlation between polymorphisms of the vitamin D receptor gene (Taq I and Fok I) and the aggressive periodontitis by FBAT method., Methods: 93 AgP nuclear families including 93 probands and their 155 relatives were recruited. The genotype frequency and polymorphism for VDR for the patients and their pedigree were detected by using polymerase chain reaction (PCR) combined with restriction fragment length polymorphism (RFLP)., Results: The frequency of allele T and t accounted for 94.6 % and 5.4 % in the total populations. No tt genotype were detected. The fathers of probands carried more allele t than the mothers(9.8% vs 1.6%, P=0.005). The frequency of allele F and f accounted for 57.1 % and 42.9 % in the total populations. The result of family based associated test (FBAT) including additive model, dominant model and recessive model showed that different alleles of Taq I and Fok I had no correlation with the onset of AgP(P>0.05)., Conclusion: This was the first time to detect the VDR gene polymorphisms in Chinese families. The result of FBAT analysis can not show the correlation between vitamin D receptor gene polymorphisms (Taq I and Fok I) and the onset of AgP.

In the title compound, C(9)H(7)ClN(2)O, the dihedral angle between the aromatic ring planes is 11.0 (2)°. In the crystal, inversion dimers linked by pairs of O-H⋯N hydrogen bonds generate R(2) (2)(8) loops.

Objective: To investigate the expression of human beta-defensin-2 (HBD-2) in laryngeal squamous cell carcinoma (LSCC), and reveal the immune significance of HBD-2 in LSCC., Methods: The expression of HBD-2 and the infiltration of CD1a(+) dendritic cells were verified by immunohistochemistry strept actividin-biotin complex (SABC) method in thirty-four cases of LSCC's paraffin sections, and the expression of HBD-2 mRNA was detected in eighteen cases of postoperative tumor specimens and five cases of normal paracarcinoma tissue using cDNA probe by in situ hybridization. Based on the type and characteristic of datas, SPSS13.0 program package was used for statistical calculation., Results: (1) The difference of expression of HBD-2 was significant among the grades of malignant cell differentiation (F = 6.809, P < 0.05), the expression of HBD-2 in well differentiated group was significantly increased compared with moderately and poorly differentiated group (P < 0.05), there wasn't significant difference with the clinical type, T status and lymphatic metastasis in the expression of HBD-2. (2) The difference of expression of HBD-2 mRNA among different grades of malignant cell differentiation was significant (F = 16.391, P < 0.05), less or no positive signal was observed in poorly differentiated group. (3) The infiltration of CD1a(+) dendritic cells had positive correlation with the expression of HBD-2 (r = 0.343, P < 0.05)., Conclusions: The expression of HBD-2 was related to the grade of malignant cell differentiation in LSCC, which was regulated at the level of transcription and translation. HBD-2 might play a role in anti-tumor immunity by chemo-attractiving and activating of immature dendritic cells.

The title compound, [Ni(C(2)Cl(3)O(2))(2)(C(5)H(5)N)(4)], was prepared by the reaction of pyridine and trichloro-acetatonickel(II) in ethanol solution at room temperature. The Ni(II) atom is located on a twofold rotation axis and has a slightly distorted octa-hedral coordination made up of four N atoms of the pyridine ligands and two O atoms of trichloro-acetate anions. The mol-ecular structure and packing are stabilized by intra- and inter-molecular C-H⋯O hydrogen-bonding inter-actions.

Background and Objective: Systemic levels of C-reactive protein and interleukin-6 have been reported to be elevated in patients with periodontitis compared with periodontally healthy individuals. Most studies included patients with chronic periodontitis and comprised predominantly Caucasians. The aim of this study was to determine the relative levels of C-reactive protein and interleukin-6 in plasma of patients with aggressive periodontitis in China and to examine the relationships between these two inflammatory mediators and clinical parameters, peripheral blood cells and protein variables., Material and Methods: Plasma samples were collected from 84 patients with aggressive periodontitis and from 65 control subjects. Periodontal examination consisted of taking probing depth and attachment loss measurements. The levels of plasma C-reactive protein and interleukin-6 were determined using enzyme-linked immunosorbent assays., Results: The levels of plasma C-reactive protein in patients with aggressive periodontitis were significantly higher than those in control subjects (1.87 vs. 0.52 mg/L). The level of plasma interleukin-6 in patients with aggressive periodontitis was 1.20 pg/mL, higher than that in control subjects (0.08 pg/mL). Multiple linear regression analysis showed that log C-reactive protein was significantly related to severe sites percentage and albumin following correction for age, gender, body mass index and smoking (p = 0.000, p = 0.008, respectively). Log interleukin-6 was found to be significantly correlated with periodontal diagnosis, leukocyte count and level of fasting blood glucose after adjusting for the confounders (p = 0.000, p = 0.009 and p = 0.013, respectively)., Conclusion: Patients with aggressive periodontitis have significantly elevated levels of plasma C-reactive protein and interleukin-6. These elevated inflammatory factors might potentially increase the risk for cardiovascular events and glucose dysregulation in relatively young individuals.

Background and Objective: The expression and activity of thioredoxin reductase (TrxR) are significantly higher in tumor tissues than in normal tissues, but the correlation of plasma TrxR activity to tumor growth has seldom been reported. This study was to evaluate the correlation of plasma TrxR activity to the growth of hepatocellular carcinoma (HCC) H22 cell xenografts in mice., Methods: H22 cells were injected subcutaneously into Kunming mice to establish HCC model. The mice were divided into control group, mice group, low dose (36 mg/kg), moderate dose (72 mg/kg) and high dose (216 mg/kg) ethaselen groups. The mice in control group and model group were given 0.5% sodium carboxymethyl cellulose (CMC-Na). Blood samples were drawn before tumor cell inoculation, when tumor volume reached 100 mm3, and at Days 1, 4 and 10 of treatment. TrxR activity in plasma and tumor tissues was detected by dithio-bis-nitrobenzoic acid (DTNB) method., Results: The inhibition rates of tumor growth were 59.5% in low dose ethaselen group, 74.3% in moderate dose ethaselen group, and 74.9% in high dose ethaselen group. Plasma TrxR activity was stable in control mice, and was correlated positively to tumor volume in tumor-bearing mice. At the end of treatment, the inhibition rates of TrxR activity in plasma and tumor were 59.2% and 15.3% in low dose ethaselen group, 68.4% and 25.8% in moderate dose ethaselen group, 68.9% and 29.8% in high dose ethaselen group., Conclusions: Plasma TrxR activity is correlated positively to tumor growth. Ethaselen can inhibit TrxR activity corresponding to tumor growth inhibition.

Background and Objective: It is known that S100A8, a member of the S100 calcium-binding protein family, is associated with inflammatory diseases, including periodontitis. Our previous population-based study found an association between two polymorphisms, rs3795391 (A > G) and rs3806232 (A > G), in the upstream region of the S100A8 gene and aggressive periodontitis (AgP) in Chinese people. Based on those results, this investigation set out to analyze and corroborate whether the association also exists within families., Material and Methods: Two hundred and four subjects from 73 nuclear families were recruited. All probands and their relatives were diagnosed according to the 1999 classification of periodontal diseases. Anticoagulated peripheral blood samples were collected from all the subjects, and DNA was extracted. The two single nucleotide polymorphisms (SNPs; rs3795391 and rs3806232) were detected and analyzed by standard polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. Analysis of genotype/allele was performed by Family-Based Association Test (FBAT) software ( http://www.biostat.harvard.edu/~fbat/default.html)., Results: There was a statistically significant association of the SNP rs3795391 with AgP in the additive genetic model (chi(2) = 3.9836, d.f. = 1, p = 0.0459). Allele A showed significantly preferential transmission to the AgP affected individuals (Z = 1.996, p = 0.0459). The other SNP, rs3806232, showed no significant results in all models., Conclusions: This family-based association study supports the previous findings that SNP rs3795391 (A > G) of the S100A8 gene might contribute to AgP susceptibility. This is, to our knowledge, the first investigation about AgP using FBAT in genetic analysis.

Cancer is a complex disease involving multiple oncogenes with diverse actions. Inhibiting only one oncogene is unlikely to eliminate the malignancy of cancer cells. The goal of this study was to investigate whether synergistic effects can be achieved by combined silencing of two oncogenes, K-ras and Akt2, which are key players in the Ras/MAPK and PI3K/Akt signaling pathways. The pancreatic cancer cell line, Panc-1, was selected for these studies as it has elevated expression of K-ras and Akt2. Compared with inhibiting each oncogene alone, simultaneously silencing the two oncogenes with RNA interference (RNAi) more effectively inhibited Panc-1 cell proliferation and colony formation, induced a significantly higher percentage of apoptosis and resulted in greater inhibition of c-myc expression in vitro. Furthermore, when delivered by polyethyleneimine into Panc-1 tumors in nude mice, RNAi simultaneously targeting K-ras and Akt2 inhibited tumor growth more efficiently than RNAi targeting the individual oncogenes. Therefore, RNAi simultaneously silencing the oncogenes K-ras and Akt2 may offer potential opportunities for pancreatic cancer gene therapy.

Objective: To investigate the clinical, radiographic and pathological characteristics of intravascular lymphomatosis primarily manifested in the lung, without skin and central nervous system involvements., Methods: A case of T cell intravascular lymphomatosis presenting with fever and multiple pulmonary shadows on chest radiograph was described and 14 similar cases reported in the English literature were reviewed., Results: We described a case of T cell intravascular lymphomatosis, who was a 36 year old man, complained of fever and multiple pulmonary shadows on chest radiograph and admitted to Peking Union Medical College Hospital in march, 2008. Open lung biopsy showed features characteristic of intravascular lymphomatosis. He received CHOP chemotherapy, but died 20 days after diagnosis. Most cases of intravascular lymphomatosis primarily manifested in the lung occurred in older patients, ranging from 36 to 79 years of age (mean 59 years), with a male predominance (M : F = 11 : 4). The chief complaints were dyspnoea (10/15), cough (9/15), fever (5/15) and weight loss (5/15). Pulmonary function tests usually revealed some degree of decreased diffusing capacity. Eight cases had high serum lactate dehydrogenase levels. Chest CT scan showed multiple reticular or/and nodular density. Immunophenotypically, 10 cases were B cell lineage, 3 cases were T cell lineage. Six cases of B cell intravascular lymphomatosis were followed, of whom 4 and were alive, and 2 died of respiratory failure. Three cases of T cell intravascular lymphomatosis showed poor prognosis, both of whom died of respiratory failure., Conclusions: Intravascular lymphomatosis primarily manifested in the lung is a rare malignant disease. Its pulmonary symptoms were nonspecific, and pulmonary function tests and chest CT scan manifested as those of interstitial pneumonia or pulmonary infection. It can be pathologically diagnosed through transbronchial lung biopsy or surgical lung biopsy.

Objective: To assess the value of color Doppler flow imaging (CDFI) in monitoring vascular complications following orthotopic liver transplantation (OLT)., Methods: Seven hundred ninety-two patients who received OLT from April 2002 to December 2006 in the Organ Transplantation Center, General Hospital of Chinese People's Armed Police Forces, Beijing, and underwent CDFI examinations in different periods after OLT were enrolled in this study. Their vascular complications were monitored by CDFI and confirmed by angiography or spiral CT., Results: Of the 792 patients, 54 were diagnosed with vascular complications that occurred 1-360 days after their OLT operations. These complications occurred within 1-30 days, 31-60 days, 61-90 days, 91-180 days, 181-360 days, with the proportions of 46.30%, 22.22%, 14.81%, 9.26% and 7.41% respectively. The proportion of hepatic artery and portal vein complications and outflow occlusions were 61.11%, 35.19% and 3.70% respectively., Conclusion: Most vascular complications occurred within six months after the OLT operation. The continuous and careful monitoring by CDFI is beneficial in an early diagnosis of vascular complications after OLT.

B3LYP/6-31G(d) hybrid HF/DFT calculations were carried out to determine the structural and electronic properties of the endohedral complex of C60 with tetrahedrane C4H4. It was demonstrated that C4H4 was seated in the center of the C60 cage and existed in a molecular form inside the fullerene. The formation of this complex was endothermic with inclusion energy of 141.05 kcal/mol. C4H4 endohedral doping slightly perturbed the molecular orbitals of C60. The calculated HOMO-LUMO gaps, the electron affinity(EA) and the ionizational potential (IP) indicated that C4H4@C60 seemed to be more kinetically reactive than C60. The IR active modes and harmonic vibrational frequencies of C4H4@C60 were also discussed.

It has been reported that the local introduction of a promoter-less DNA containing the complementary DNA (cDNA) sequence of a gene could induce gene-specific silencing in plants. The feasibility of this kind of silencing in human cancer cells is as yet unknown. The current study was designed to investigate the anti-tumor effects of a homologous promoterless K-ras cDNA system on pancreatic cancer. A full-length K-ras cDNA fragment was cloned into the promoterless plasmid puc19 to yield puc-K-ras. This construct was then transfected into pancreatic cancer cells. Our results demonstrated that the transfection of a promoterless K-ras cDNA resulted in a significant decrease in endogenous K-ras in a dose- and time-dependent manner and induced pancreatic cell apoptosis. Furthermore, stable puc-K-ras transfection decreased the endogenous protein level of K-ras and inhibited cell proliferation, clone formation and tumorigenicity in vivo. These findings indicate a promising application of this homologous promoterless cDNA silencing system in pancreatic cancer gene therapy.

The title compound, C(14)H(12)N(2)O(3), was prepared by reaction of 2-nitro-benzaldehyde with 4-methoxy-benzenamine at 377 K. The molecule has an E configuration, with a dihedral angle between the two benzene rings of 43.3 (5)°. An intermolecular C-H⋯O interaction links molecules in zigzag chains down the a axis.

Oxidative stress, especially peroxynitrite (ONOO(-))-mediated oxidative stress, plays a key role in diabetes. Mitochondria, as the generating source of ONOO(-), may also be the major damaging target of ONOO(-), which can cause a series of mitochondrial proteins nitration. Therefore, this study aimed to clarify the relationship between the nitration of entire mitochondrial proteins induced by ONOO(-) and liver mitochondrial structural damage in diabetes. Sprague-Dawley male rats were injected with streptozotocin to induce diabetes. After 10 weeks, transmission electron microscopy was used to observe the ultrastructure of liver mitochondria, and reverse transcription-polymerase chain reaction was used to detect liver inducible nitric oxide synthase (iNOS) mRNA expression. Nitrotyrosine (NT) content and distribution were detected with Western blot analysis and immunohistochemistry. In addition, some biochemical indicators were detected to represent oxidative stress and metabolic disorders. In diabetic rats, increasing levels of iNOS mRNA and NT content (P<.05) were observed, in accord with pathological alterations of the ultrastructure of liver mitochondria. Meanwhile, some alterations in biochemical indicators were observed in diabetes. Treatment with aminoguanidine could significantly attenuate these alterations (P<.01 or P<.05). In conclusion, the nitration of mitochondrial proteins induced by ONOO(-) may be responsible for structural damage to liver mitochondria, and aminoguanidine can reduce ONOO(-) generation and attenuate mitochondrial damage.

The title compound, C(13)H(10)BrNO, was prepared by reaction of 3-bromo-aniline with 2-hydroxy-benzaldehyde at 377 K. The mol-ecular structure and packing are stabilized by an intra-molecular O-H⋯N hydrogen-bond inter-action.

The crystal structure of the title compound, C(7)H(8)N(5)S(+)·Cl(-), is stabilized by inter-molecular N-H⋯Cl and N-H⋯S hydrogen-bond inter-actions.

The title compound, C(14)H(12)ClNO, was prepared by the reaction of 4-methoxy-aniline and 4-chloro-benzaldehyde in ethanol at 367 K. The mol-ecule is almost planar, with a dihedral angle between the two benzene rings of 9.1 (2)° and an r.m.s. deviation from the mean plane through all non-H atoms in the mol-ecule of 0.167 Å.

Objective: To study whether interleukin-1( IL-1 )genotype and/or alleles were associated with aggressive periodontitis (AgP) patients and their relatives., Methods: Sixty-six AgP nuclear families including 66 probands and their 114 relatives were recruited. All probands were diagnosed as AgP according to 1999 classification of periodontal diseases and conditions, the diagnoses of their relatives were using a system of assessment (including the current and retrospective diagnosis and reported case history). Anti-coagulated peripheral blood samples were collected from all of the 180 subjects, and DNA was extracted from each blood sample. Single nucleotide polymorphisms (SNPs) at IL-1A +4845 and IL-1B j 511 were analyzed by PCR-RFLP assay. The polymorphism of a variable number of tandem repeat (VNTR) in intron 2 of IL-1RN was detected by PCR amplification and fragment size analysis. The distributions of genotypes and/or alleles in all groups were analyzed by logistic regression analysis. Data were also analyzed using the family-based association test., Results: The frequency of all markers (at IL-1A +4845 and IL-1B -511 and VNTR) showed no association wih AgP in relatives (P>0.05). Tests of association by Family-Based Association Test showed no difference in the distribution of the allele 1 of IL-1B-511 under all three genetic models tested (Additive: Z=0.389, P=0.6969; Dominant: Z=0.089, P=0.9287; Recessive: Z=0.089, P=0.9287). The distribution of the allele 1 of IL- 1A+4845 also showed no significant difference under all three models (P>0.05). But when all AgP patients were stratified by the clinical subtype, the frequencies of A1A1 genotype and allele 1 at IL-1B-511 were significantly increased in the localized subtype AgP patients compared to unaffected (genotype: 43.8% vs 22.8%,OR=7.32,95% CI=1.84-29.11,P=0.005; allele 1: 71.9% vs 41.2%,OR=3.64,95% CI=1.55-8.58,P=0.002), while no significant difference was observed between the generalized subtype and the unaffected (genotype: P=0.88, allele: P=0.64)., Conclusion: The polymorphism of IL-1B-511 may have an effect on the susceptibility of localized AgP in Chinese population. It should be tested in larger family samples.

In the title compound, C(20)H(17)N(3), there are four mol-ecules in the asymmetric unit. The dihedral angles between the indolizine ring system and the pyridyl ring are 6.6 (2), 7.4 (1), 4.0 (1) and 10.1 (4) in the four molecules. There are no further important differences between the independent molecules. In each molecule, there is an intra-molecular C-H⋯N hydrogen bond. The whole structure is stabilized by N-H⋯π and C-H⋯π inter-actions.

In the title compound, C(12)H(12)N(2)O(2)S, there are S-H⋯N and N-H⋯O hydrogen-bond inter-actions. The N-H⋯O hydrogen bond is bifurcated, with the hydrogen being simultaneously donated to two equivalent O atoms, forming one intra- and one inter-molecular N-H⋯O bond with an R(1) (2)(4) motif. The motif of the S-H⋯N hydrogen bond is R(2) (2)(12).

Chromosomal coexpression domains are found in a number of different genomes under various developmental conditions. The size of these domains and the number of genes they contain vary. Here, we define local coexpression domains as adjacent genes where all possible pair-wise correlations of expression data are higher than 0.7. In rice, such local coexpression domains range from predominantly two genes, up to 4, and make up approximately 5% of the genomic neighboring genes, when examining different expression platforms from the public domain. The genes in local coexpression domains do not fall in the same ontology category significantly more than neighboring genes that are not coexpressed. Duplication, orientation or the distance between the genes does not solely explain coexpression. The regulation of coexpression is therefore thought to be regulated at the level of chromatin structure. The characteristics of the local coexpression domains in rice are strikingly similar to such domains in the Arabidopsis genome. Yet, no microsynteny between local coexpression domains in Arabidopsis and rice could be identified. Although the rice genome is not yet as extensively annotated as the Arabidopsis genome, the lack of conservation of local coexpression domains may indicate that such domains have not played a major role in the evolution of genome structure or in genome conservation.

Objective: To investigate whether introducing promoterless DNA containing the cDNA sequence of green fluorescent protein (GFP) induces gene-specific silencing in human pancreatic cancer cell line harboring genomically integrated GFP gene., Methods: Using G418 selection and fluorescent separation we established a highly purified monoclonal pancreatic cancer cell line, recombinant PANC-1, which had a steady level of GFP expression. GFP cDNA was amplified by PCR from plasmid pEGFP-C1 and ligated to the promoterless plasmid PUC19. PUC-GFP was then transfected into monoclone cells along or cotransfected with pEGFP-C1 to panc-1 cells. Each had PUC plasmid transfected group as their control to eliminate possibility of plasmid toxicity and GFP small interferent RNA (siGFP) transfected group as positive control. Western blot, flow cytometry and phase contrast fluorescence microscopy were used to detect the changes of GFP expression., Results: The data showed that (1) PUC-GFP inhibited the GFP expression in monoclonal cell line in a dosage dependent manner. The inhibitory effect of 3 microg PUC-GFP did not show significant difference with siGFP. (2) The significant repression appeared on the fourth day after transfecting monoclonal cells with 3 microg PUC-GFP. By the end of the sixth day, GFP expression in PUC-GFP group and siGFP group remained at a low level. (3) Cotransfecting PUC-GFP with pEGFP-C1 plasmids into PANC-1 cells showed a decreased transfection efficiency when compared with transfecting pEGFP-C1 alone. Higher PUC-GFP vs pEGFP-C1 corresponded with lower transfection efficency. (4) When adding new pEGFP-C1 plasmid to cells after inhibition appeared, the GFP expression recovered., Conclusion: Transfection of the promoterless DNA fragment containing full-length cDNA effectively induces a gene-specific silencing in mammalian cells.

Ab initio Hartree-Fock (HF) and density functional theory (DFT) calculations were carried out to determine the structural and electronic properties of the highest epoxygenated fullerenes C(60)O(30). For comparison, other fullerene oxides C(60)O(29), C(60)O(3), C(60)O(2) and C(60)O were also studied. The highly symmetrical I(h) structure of the parent C(60) is reserved in C(60)O(30) and C(60)O(30) was calculated to be a nonpolar molecule. It was demonstrated that C(60)O(30) should be more stable than other C(60) oxides such as C(60)O(29), C(60)O(3), C(60)O(2) and C(60)O. Compared with C(60), it is less possible for C(60)O(30) to accept or donate electrons from the reduced EAs and enhanced IPs. The IR active modes and harmonic vibrational frequencies of C(60)O(30) were also discussed.

Tetramethylpyrazine (TMP), an effective component of traditional Chinese medicine Chuanxiong, is commonly used to resolve embolism. Its possible therapeutic effect against atherosclerosis has received considerable attention recently. Angiotensin II (Ang II) is highly implicated in the proliferation of vascular smooth muscle cells (VSMCs), resulting in atherosclerosis. The mechanisms of TMP in the proliferation of VSMCs induced by Ang II remain to be defined. The present study was aimed to study the effect of TMP on Ang II-induced VSMC proliferation through detection of nuclear factor-kappaB (NF-kappaB) activity and bone morphogenetic protein-2 (BMP-2) expression. Primary cultured rat aortic smooth muscle cells were divided into the control group, Ang II group, Ang II + TMP group and TMP group. Cells in each group were harvested at different time points (15, 30 and 60 min for detection of NF-kappaB activity; 6, 12 and 24 h for measurement of BMP-2 expression). NF-kappaB activation was identified as nuclear staining by immunohistochemistry. BMP-2 expression was observed through Western blot, immunohistochemistry and in situ hybridization. The results showed that: (1) Ang II stimulated the activation of NF-kappaB. Translocation of NF-kappaB p65 subunit from cytoplasm to nucleus appeared as early as 15 min, peaked at 30 min (P<0.01) and declined after 1 h. (2) TMP inhibited Ang II-induced NF-kappaB activation (P<0.01). (3) Ang II increased BMP-2 expression at 6 h but declined it significantly at 12 and 24 h (P<0.01). (4) BMP-2 expression was also kept at high level at 6 h in Ang II + TMP group but maintained at the normal level at 12 and 24 h. (5) There was no significant difference in NF-kappaB activation and BMP-2 expression between the control group and TMP group. These results indicate that TMP inhibits Ang II-induced VSMC proliferation through repression of NF-kappaB activation and BMP-2 reduction, and BMP-2 expression is independent of the NF-kappaB pathway. In conclusion, TMP has therapeutic potential for the treatment of atherosclerosis.

Objective: To explore the factors influencing the clinical therapeutic effect of micro-invasion catgut embedding therapy., Methods: The authors attain preliminary understanding from their long term clinical studies on micro-invasion catgut embedding therapy, especially, exploration and recognition, in combination with clinical practice., Conclusion: Theories of channels, and reinforcing-reducing manipulation of embedding thrust, the catgut-embedded depth, selection of the thrust, cycle of treatment, diet regulation and selection of indications, etc. are key factors of influencing therapeutic effect of the micro-invasion catgut embedding therapy.

Two new labdane diterpenes, 14(R),15-dihydroxy-8(17),12(E)-labdadien-19-oic acid (1) and 16-methyl-12,15-epoxy-8(17),13-labdadien-19-oic acid (2), together with four known compounds, were isolated from the seeds of Platycladus orientalis. Their structures were established by spectroscopic methods. The stereochemistry of compound 1 was defined by X-ray crystallographic analysis.

Aim: To investigate the effect of small interfering RNAs targeting mutant K-ras on the growth of pancreatic carcinoma cell lines in vitro and in vivo., Materials and Methods: We cloned targeting sequence spanning codon 12 of mutant K-ras into the pSilencer-hygro plasmid, yielding two recombinant vectors with one base different. Both human pancreatic carcinoma cell lines were transfected by these two recombinant vectors. The transfected PC-7 cells were injected subcutaneously into nude mice to observe its tumorigenicity. RT-PCR and Western blot analysis were carried out to test the expression of K-ras in all of the transfected cell lines. Growth curves assay were performed to test the abilities of cells proliferation. Anti-K-ras therapy of PC-7 and Panc-1 in subcutaneous mice models were performed by intratumor injection of polyethylenimine/siRNAs complex., Results: The expressions of K-ras in PC-7 cells and Panc-1 cells were significantly inhibited by corresponding small interfering RNAs. The expression of K-ras was particularly inactivated by siRNA without any base mismatch to its homologous mRNA, while this oncogene with central base mismatch could not be inhibited as effectively as that of the former. The growth of PC-7 cells and Panc-1 cells transfected by corresponding mutant K-ras targeted siRNAs were significantly suppressed when compared with controls (p<0.05). The transfected PC-7 cells lost tumorigenic ability. Four weeks treatment of Xenograft of pancreatic carcinoma (PC-7 and Panc-1) in nude mice with Polyethylenimine-encapsulated mutant K-ras targeted siRNAs (20 microg/mouse twice weekly) were effective in reducing tumor growth, when compared with controls (p<0.05)., Conclusion: The central base may play a key role in the process of RNA interference. The mutant point and its vicinity of 19 nucleotides in K-ras may be the effective targeting sequence for RNA interference. Targeting mutant-k-ras therapy of pancreatic carcinoma may be a clinically applicable therapeutic modality.

Objective: To investigate the roles of K-ras gene in the tumorigenesis of ovarian serous borderline and malignant tumors., Methods: Fifty one tissue samples of ovarian serous tumors, including 18 conventional serous borderline tumors, 11 micropapillary serous borderline tumors, 12 invasive micropapillary serous carcinomas, and 10 conventional serous carcinomas were investigated for the presence of K-ras mutation. DNA was extracted after microdissection of the tumor tissue, the exon 1 of K-ras gene was amplified by PCR, and the presence of mutation at the codons 12 and 13 was evaluated by direct sequencing analysis., Results: GGT to GTT mutation at codon 12 of the K-ras gene was found in one conventional serous borderline tumors, resulting in valine to glycine substitution. All other 50 cases showed no K-ras mutation. All tumors had a wild-type codon 13., Conclusions: Mutations of K-ras at codons 12 and 13 in ovarian serous tumors are very rare in this series of patients, suggesting a difference present between the Chinese and Caucasian populations. K-ras mutations may play a less important role in the tumorigenesis of ovarian serous tumor of the Chinese patients.

In both the monocot rice and the dicot Arabidopsis, highly expressed genes have more and longer introns and a larger primary transcript than genes expressed at a low level: higher expressed genes tend to be less compact than lower expressed genes. In animal genomes, it is the other way round. Although the length differences in plant genes are much smaller than in animals, these findings indicate that plant genes are in this respect different from animal genes. Explanations for the relationship between gene configuration and gene expression in animals might be (or might have been) less important in plants. We speculate that selection, if any, on genome configuration has taken a different turn after the divergence of plants and animals.

Background: White Spot Syndrome Virus, a member of the virus family Nimaviridae, is a large dsDNA virus infecting shrimp and other crustacean species. Although limited information is available on the mode of transcription, previous data suggest that WSSV gene expression occurs in a coordinated and cascaded fashion. To search in silico for conserved promoter motifs (i) the abundance of all 4 through 8 nucleotide motifs in the upstream sequences of WSSV genes relative to the complete genome was determined, and (ii) a MEME search was performed in the upstream sequences of either early or late WSSV genes, as assigned by microarray analysis. Both methods were validated by alignments of empirically determined 5' ends of various WSSV mRNAs., Results: The collective information shows that the upstream region of early WSSV genes, containing a TATA box and an initiator, is similar to Drosophila RNA polymerase II core promoter sequences, suggesting utilization of the cellular transcription machinery for generating early transcripts. The alignment of the 5' ends of known well-established late genes, including all major structural protein genes, identified a degenerate motif (ATNAC) which could be involved in WSSV late transcription. For these genes, only one contained a functional TATA box. However, almost half of the WSSV late genes, as previously assigned by microarray analysis, did contain a TATA box in their upstream region., Conclusion: The data may suggest the presence of two separate classes of late WSSV genes, one exploiting the cellular RNA polymerase II system for mRNA synthesis and the other generating messengers by a new virus-induced transcription mechanism.

Objectives: To explore the diagnostic value of color Doppler flow imaging (CDFI) in monitoring portal vein complications (PVC) following orthotopic liver transplantation (OLT)., Methods: Five hundred and four patients received OLT and had CDFI examinations before and after their transplantations. CDFI monitoring parameters before the operation included portal vein diameter, blood flow velocity and the presence of thrombi within the portal vein. Monitoring parameters after the operation included portal vein diameters of donor and receiver sides, and the diameter of the mouth of anastomosis, inner side of blood vessel echo, the direction of blood flow and its speed., Results: Of the 504 patients examined, the median velocity of the portal blood flow was 46.27 cm/s (range: 15.8 to 110.8) on the day of the operation. The blood flow speed of 358 cases (71.03%) was higher than 40 cm/s. Of the 358 patients, 347 (68.85%) had a blood flow speed lower than 40 cm/s one month after the operation. Sixty-four patients (12.70%) showed hepatofugal blood flow once, and only one case had a total hepatofugal blood flow. Thirteen patients (2.58%) had portal vein complications, including 4 cases with portal vein thrombosis and 9 with portal vein stenosis., Conclusion: Not all abnormal portal vein blood flow signals will lead to complications. It is worthwhile to monitor the portal blood flows. CDFI plays an important role in the diagnosis of portal vein complications after orthotopic liver transplantation.

Expression of genes in eukaryotic genomes is known to cluster, but cluster size is generally loosely defined and highly variable. We have here taken a very strict definition of cluster as sets of physically adjacent genes that are highly coexpressed and form so-called local coexpression domains. The Arabidopsis (Arabidopsis thaliana) genome was analyzed for the presence of such local coexpression domains to elucidate its functional characteristics. We used expression data sets that cover different experimental conditions, organs, tissues, and cells from the Massively Parallel Signature Sequencing repository and microarray data (Affymetrix) from a detailed root analysis. With these expression data, we identified 689 and 1,481 local coexpression domains, respectively, consisting of two to four genes with a pairwise Pearson's correlation coefficient larger than 0.7. This number is approximately 1- to 5-fold higher than the numbers expected by chance. A small (5%-10%) yet significant fraction of genes in the Arabidopsis genome is therefore organized into local coexpression domains. These local coexpression domains were distributed over the genome. Genes in such local domains were for the major part not categorized in the same functional category (GOslim). Neither tandemly duplicated genes nor shared promoter sequence nor gene distance explained the occurrence of coexpression of genes in such chromosomal domains. This indicates that other parameters in genes or gene positions are important to establish coexpression in local domains of Arabidopsis chromosomes.

We analyze a method for efficiently generating optical vortices by use of annular computer-generated holograms and a spatial light modulator. We found that there exists an optimal annular width by which the reconstructed vortex ring in the focal plane has the steepest gradient and the worthless subbright rings can be largely suppressed. We fitted a general formula for determining the value of this optimal annular width and propose a method for designing a multiring structure of optical vortices and specialized interferometric vortex patterns. Finally, we discuss the situation of a Gaussian beam as illuminated light and find that there exists an optimal beam waist that results in the best energy efficiency.

Structural and electronic properties of S-doped fullerene C58 were calculated systematically via Hartree-Fock self-consistent field (SCF) and density functional B3LYP levels of theory with 6-31G(d) basis set. The most stable C58S represents an open cage structure with a nine-member ring orifice, which provides a large hole for large atoms or small molecules to pass through into the cage. The most stable endohedral S@C58 has the S atom seated near the center of the C58 cage. The calculated highest occupied molecular orbital-lowest unoccupied molecular orbital energy gaps of the isomers lie in the range of 1.42-2.50 eV. The electron affinity and the ionization potential were also presented as an indicator of the kinetic stability. Our results may aid in the design of experimental methods for controlling the nature of fullerene cages (for example, doping, opening, and reclosing them).

We report a new optical vortex phase-shifting method for digital holography, in which an optical vortex mode is taken as the reference beam for holographic recording, and the required phase shifts are directly generated by rotating the vortex mode. In digital reconstruction, the complex amplitude of the object wave can be retrieved by use of the conventional phase shifting algorithm on condition that the digital illumination beam is replaced by an vortex beam with the same topological charge as the reference used. Both the theoretical analysis and experimental results demonstrate the feasibility of this approach.

Objective: To investigate the toxicity of alpha-terthienyl to the larvae of Aedes albopictus, its influencing factors and effect on the larva development., Methods: Under experimental ultraviolet-A (UVA), the number of dead,pupal or eclosive mosquito larvae was determined on the condition of different doses of alpha-terthienyl and different disposal time in the dark; the number of dead larvae was also determined under sunlight on the condition of different doses of alpha-terthienyl and different disposal time to water., Results: The LC50 of alpha-terthienyl to Aedes albopictus larvae was 2.37 microg/L under UVA. The best effect was shown when the larvae were incubated with alpha-terthienyl 3 h in dark. Alpha-terthienyl could significantly inhibit the larva development and the emergence of the pupae. Under strong sunlight, the larvae were quickly killed by high concentration alpha-terthienyl. The 24 hours effect of alpha-terthienyl was better when it was applied at 5 AM than that of at 10 AM and 1 PM., Conclusion: Alpha-terthienyl is an effective, practicable larvicide which prohibits the growth and development of the larvae of Aedes albopictus.

Purpose: The goal of this study was to explore the relationship between feeding methods and deciduous caries., Methods: An investigation on children's feeding methods, caries prevalence and dmft was made, using 260 cases which were provided with whole primary teeth. Chi-square test was used for statistical analysis on feeding methods and caries prevalence, and variance analysis was used on feeding methods and dmft., Results: The results showed that the deciduous caries rate, mean dmft in breast feeding (BF) group were lower than those in mixed feeding (MF) group and artificial feeding (AF) group, there were significant differences between BF group and MF group,also between BF group and AF group. But there were no significant differences between MF group and AF group., Conclusion: It is concluded that proper breast feeding seems to be important for preventing deciduous caries.

A new simple approach basing on the histogram and genetic algorithm(GA) to efficiently detect QRS-T complexes of the ECG curve is described, so as to easily get the P-wave (when AF does not happen) or the f-wave (when AF happens). By means of signal processing techniques such as the power spectrum function, the auto-correlation function and cross-correlation function, two kinds of ECG signal when AF does or does not happen were successively analyzed, showing the evident differences between them.

Visual Evoked potentials (VEPs) are time-varying signals typically buried in relatively large background noise known as the electroencephalogram (EEG). An adaptive noise cancellation with neural-network-based fuzzy inference system was used and the NNFIS was carefully designed to model the VEP signal. An advantage of the method in this paper is that no reference signal is required. The NNFIS based on Takagi and Sugeno's fuzzy model has the advantage of being linear-in-parameter, which is able to closely fit any function mapping and can track the dynamic behavior of VEP in a real-time fashion. 4 sets of simulated data indicate that the proposed method is appropriate to estimate VEP. A total of 150 trials are processed to demonstrate the superior performance of the proposed method.

Aim: Rodent testes are generally more susceptible to cadmium (Cd)-induced toxicity than liver. To clarify the molecular mechanism of Cd-induced toxicity in testes, we compared metallothionein (MT) gene expression, MT protein accumulation, and Cd retention at different time in freshly isolated testicular interstitial cells and liver of rats treated with Cd., Methods: Adult male Sprague-Dawley rats weighing 250-280 g received a s.c injection of 4.0 micromol Cd/kg and were euthanized by CO(2) asphyxiation 1 h, 3 h, 6 h, or 24 h later. Tissue was sampled and testicular interstitial cells were isolated. There were three replicates per treatment and 3 animals per replicate for RNA analyses, others, three replicates per treatment and one animal per replicate. MT1 and MT2 mRNA levels were determined by semi-quantitative RT-PCR analysis followed by densitometry scanning, and MT was estimated by the enzyme-linked immunosorbent assay (ELISA) method. Cadmium content was determined by atomic absorption spectrophotometry. The same parametersd were also analyzed in the liver, since this tissue unquestionably accumulate MT., Results: The rat testis expressed MT1 and MT2, the major isoforms. We also found that untreated animals contained relatively high basal levels of both isoform mRNA, which were increased after Cd treatment in liver and peaked at 3 h, followed by a decline. In contrast, the mRNA levels in interstitial cells peaked at 6 h. Interestingly, the induction of MT1 mRNA was lower than MT2 mRNA in liver of rat treated with Cd, but it was opposite to interstitial cells. Cd exposure substantially increased hepatic MT (3.9-fold increase), but did not increase MT translation in interstitial cells., Conclusion: Cd-induced expression of MT isoforms is not only tissue dependent but also time-dependent. The inability to induce the metal-detoxicating MT-protein in response to Cd, may account for a higher susceptibility of testes to Cd toxicity and carcinogenesis compared to liver.

The rodent testes are generally more susceptible to cadmium (Cd)-induced toxicity than the liver. To clarify the molecular mechanism underlying tissue and cell differences in Cd sensitivity, we compared metallothionein (MT) gene expression, MT protein accumulation, and Cd retention under different times in freshly isolated testicular Sertoli and spermatogenic cells and liver of rats treated with Cd. Adult male Sprague-Dawley rats received a s.c. injection of 4.0 micromol Cd/kg and 1, 3, 6, or 24h later and untreated animals (0h) tissue were sampled and testicular Sertoli and spermatogenic cells isolated. MT1 and MT2 mRNA levels were determined by semi-quantitative RT-PCR analysis followed by densitometry scanning, and MT was estimated by the enzyme-linked immunosorbent assay (ELISA) method. Cadmium content was determined by atomic absorption spectrophotometry. Testicular lesions were not grossly or histologically observed in rats treated with 4.0 micromol Cd/kg. In the present study, we demonstrated that the rat testis indeed expressed MT1 and MT2, the major isoforms. We also found that untreated animals contained relatively high basal levels of both isoform mRNA, which were increased after Cd treatment in liver and peaked at 3h, followed by a decline, in contrast, the mRNA levels in Sertoli cells peaked at 6h. Interestingly, the induction of MT1 mRNA was lower than MT2 mRNA in Sertoli cells and liver of rats treated with Cd. However, the MT1 mRNA levels of spermatogenic cells decreased 0-3h after Cd treatment, followed by an increase; in contrast, MT2 mRNA levels increased 0-3h after Cd treatment, followed by a reduction, but induced extents of them are lower than those of Sertoli cells and liver. Cd exposure substantially increased hepatic MT, but did not increase MT translation in Sertoli and spermatogenic cells. These results indicate: (1) that Cd-induced MT mRNA expression is cell- and time-dependent; (2) that the inability to induce the metal-detoxicating MT protein in response to Cd, might account for higher susceptibility of testes to Cd toxicity and carcinogenesis relative to liver.