PURPOSE: The aim of the present study was to investigate the prognostic value of tumor-infiltrated lymphocytes (TILs), especially the prognostic value of Foxp3+ regulatory T cells (Tregs), CD8+ CTLs and Tregs/CD8+ ratios in gastric cancer patients after R0 resection. PATIENTS AND METHODS: From 133 patients, CD4+, CD8+ and Foxp3+ TILs were assessed by immunohistochemistry in tissue microarrays and N1 regional lymph nodes sections containing gastric cancer. The prognostic effects of low- or high-density TIL subsets were evaluated by Cox regression and Kaplan-Meier analysis using median values as cutoff, while the effects of Foxp3+/CD8+ ratios were evaluated using the value determined by ROC cure analysis as cutoff. RESULTS: It was found that CD4+ and CD8+ TILs were not associated with overall survival (OS). In the tumor sites, higher Foxp3+ Tregs/CD8+ ratio was an independent factor for worse OS (multivariate analysis HR = 2.827, P = 0.037). The 1-year, 2-year and 3-year OS rates were 90, 77.5 and 70% for the group with intratumoral high Tregs/CD8+ ratio, compared with 100, 94.3 and 90.5% for the group with intratumoral low ratio. At the same time, the presence of intratumoral high Foxp3+ Tregs was also associated with worse OS (log rank test, P = 0.025); however, it was not an independent predictor and correlated with intratumoral Foxp3+ Tregs/CD8+ ratio (chi (2) test, P < 0.001). Although the infiltration of Foxp3+ Tregs in N1 regional lymph nodes was associated with lymph node metastasis (P = 0.028), it was not associated with prognosis (P = 0.458). CONCLUSIONS: Intratumoral high Foxp3+ Tregs/CD8+ ratio was an independent predictor for the prognosis of gastric cancer. It can be inferred that a combination of deletion of Tregs and stimulation of CD8+ effector T cells may be an effective immunotherapy to prolong survival after surgery.

A Gram-positive anaerobic pathogen, Clostridium perfringens, causes clostridial myonecrosis or gas gangrene in humans by producing numerous extracellular toxins and enzymes that act in concert to degrade host tissues. The ability of infectious bacteria to acquire sufficient iron during infection is essential for the pathogen to cause disease. In the C. perfringens strain 13 genome, a heme oxygenase gene homologue (CPE0214, hemO) was found and its role was examined. The purified recombinant HemO protein showed heme oxygenase activity that can convert heme to biliverdin. hemO transcription was induced in response to extracellular hemin in a dose-dependent manner. The global two-component VirR/VirS regulatory system and its secondary regulator VR-RNA had positive regulatory effects on the transcription of hemO. These data indicate that heme oxygenase may play important roles in iron acquisition and cellular metabolism, and that the VirR/VirS-VR-RNA system is also involved in the regulation of cellular iron homeostasis, which might be important for the survival of C. perfringens in a human host.

The pathogenesis of tuberculosis causing Mycobacterium bovis is largely due to its successful entry and survival in macrophages. Previous research indicated that mycobacteria-specific PE_PGRS genes code for cell surface proteins which may have role in mediating interactions with macrophages. In this study, we expressed PE_PGRS 62 gene in a non-pathogenic fast growing Mycobacterium smegmatis strain and found that the recombinant Mycobacterium smegmatis decreased macrophages livability in a dosage-dependent manner and time-dependent manner, compared with parental strain containing the vector only. To explore whether PE_PGRS 62 modulates the gene expression profile of macrophages, we stimulated macrophages by the M. smegmatis strain expressing PE_PGRS 62 as well as the control strains, followed by real-time RT-PCR assay for the mRNA expression level of IL-1beta, IL-6, and iNOS. The results showed that the expression of IL-1beta, IL-6 in macrophages were down-regulated by stimulation with the M. smegmatis strain expressing PE_PGRS 62 compared to the control strains (P < 0.05). In contrast, there were no measurable differences in the expression of iNOS. Overall, we demonstrated that PE_PGRS 62 protein altered the immune environment of the host cells, which suggest that the pathogenic PE_PGRS 62 protein altering the immune mechanism maybe involved in the pathogenesis of mycobacterial disease.

Nanoscaled AgO with high purity was synthesized by a chemical oxidation method. It can perform well as the cathode material in the proposed Li-AgO battery which coupled a Li metal anode and employed an aqueous//LISICON//nonaqueous electrolyte. The assembled Li-AgO cell demonstrated both a higher energy density and a superior power performance than its predecessors.

Xanthate was successfully grafted onto bentonite by a relatively simple solution reaction. The obtained xanthated bentonite (XBent) was characterized by FT-IR spectrophotometer, thermogravimetric analysis (TG), particle size analysis, x-ray diffraction (XRD) and scanning electron microscopy (SEM). XBent acting as a type of environmentally friendly adsorbent was applied to remove lead ions from aqueous solutions. The optimum conditions were as follows: [Pb(2 + )]=500 mg L(-1), [XBent]=2 g L(-1), pH = 5.0; oscillating 60 min under 200 rpm at 25 degrees C. The removal rate of lead was up to 99.9%. It was found that the lead(II) ions-XBent adsorption isotherm model fitted well to the Freundlich isotherm. The adsorption mechanism was also investigated by SEM and XRD, which concluded that lead ions were complexed or chelated with XBent. XBent appears to have potential to be used later in water treatment as a type of inorganic polymer reagent.

L-selectin is an adhesion molecule expressed by neutrophils that broadly directs their infiltration in to sites of inflammation. It is also present at relatively high levels in the serum of normal individuals. It is well established that L-selectin is efficiently shed from the surface of neutrophils upon their activation, a process that regulates its density and binding activity. Neutrophil programmed cell death is critical for the resolution of inflammation, and L-selectin downregulation is induced during this process as well. The mechanisms underpinning this latter process are much less understood, and were investigated in this study. Using a disintegrin and metalloprotease (ADAM)-17 radiation chimeric mice, we demonstrate for the first time that during early events of death receptor-mediated neutrophil apoptosis, L-selectin downregulation occurs primarily by ADAM17-mediated shedding. This was observed as well upon using shRNA to knock down ADAM17 expression in Jurkat cells, a well-studied cell line in terms of the molecular processes involved in the induction of apoptosis. These findings directly reveal that ADAM17 activity occurs during programmed cell death. Hence, the cleavage of particular ADAM17 substrates may be an additional component of the anti-inflammatory program initiated by apoptotic neutrophils. Of interest was that during later stages of induced leukocyte apoptosis, soluble L-selectin production occurred independent of ADAM17, as well as membrane events, such as blebbing and microparticle production. This process may provide an explanation for the lack of diminished serum L-selectin levels in ADAM17-null mice, and suggests a mechanism for the homeostatic maintenance of soluble L-selectin levels in the blood of healthy individuals.

A new support matrix inspired by honeycomb was developed for cell immobilization to control fungal morphology and enhance mass transfer in bioreactor for lactic acid production by Rhizopus oryzae. The immobilization matrix composed of asterisk-shaped fibrous matrices in a honeycomb configuration provided high surface areas for cell attachment and biofilm growth. More than 90% of inoculated spores were adsorbed onto the matrices within 6-8h and after 10h there was no suspended cell in the fermentation broth, indicating a 100% immobilization efficiency. Compared to free-cell fermentation, lactic acid production increased approximately 70% (49.5g/L vs. 29.3g/L) and fermentation time reduced 33% (48h vs. 72h) in shake-flasks with 80g/L initial glucose. The immobilized-cell fermentation was evaluated for its long-term performance in a bubble-column bioreactor operated in a repeated batch mode for nine cycles in 36days. The highest lactic acid production was 68.8g/L, corresponding to a volumetric productivity of 0.72g/Lh and 93.4% (w/w) lactic acid yield from consumed glucose. The overall yield and productivity were 77.6% and 0.57g/Lh, respectively. The fermentation can be improved by increasing aeration and mixing in the bubble-column bioreactor.

BACKGROUND: The Mainland Chinese version of the Edinburgh Postnatal Depression Scale (EPDS) has been validated among antenatal women, subsequent validation among postnatal women is necessary. OBJECTIVES: This study investigated the prevalence of postnatal depression and the psychometric characteristics of the Mainland Chinese version of the EPDS among postnatal women. DESIGN: Two studies were used for validating the EPDS with specific aims. Study I established the psychometric properties of the EPDS by examining the convergent, discriminant and construct validity, internal consistency and stability of the scale. Study II established its sensitivity, specificity and the optimal cut-off score of the EPDS according the DSM-IV-TR criteria using the Structured Clinical Interview. SETTING: : Four regional public hospitals in Chengdu. PARTICIPANTS: : A convenience sample was composed of 300 and 342 postnatal women in study I and II, respectively. METHOD: A receiver operating characteristic (ROC) analysis was carried out to evaluate the global functioning of the scale. The Beck Depression Inventory (BDI), Dyadic Adjustment Scale (DAS) and standard SF-12 Health Survey (SF-12) were used to investigate the convergent, discriminant and construct validity. An exploratory factor analysis was used to investigate the structural validity of the scale. Confirmatory factor analysis (CFA) was tested the proposed factor module by Linear Structural Relations (LISREL). The Cronbach's alpha reliability coefficient, split-half reliability and test-retest reliability were used to examine the internal consistency and stability of the scale. RESULTS: The prevalence of postnatal depression was 4.7%. Structural validity revealed a three-factor structure for the EPDS and the CFA showed a good overall fit of this three-factor model. Convergent and construct validity was supported and discriminant validity suggested that the EPDS successfully discriminated among the nondepressed, mildly and clinically depressed groups. The area under curve (AUC) was 89.6% and the logistic estimate for the threshold score was 10.5 (sensitivity, 81.25%; specificity, 80.67%) for clinical depression. The split-half reliability of the EPDS was 0.74, Cronbach's alpha was 0.78 and test-retest reliability was 0.90. CONCLUSIONS: Our data confirm the validity of the Mainland Chinese version of EPDS in identifying postnatal depression among postnatal women in Chengdu by using >10 cut-off point. Because of its brevity and acceptability, it is recommended that the EPDS be used in routine postnatal screening.

Yes-associated protein (YAP), the nuclear effector of the Hippo pathway, is a key regulator of organ size and a candidate human oncogene. This study aimed to assess the clinical significance and biological functions of YAP in non-small-cell lung cancer (NSCLC). We investigated the expression of YAP in 92 cases of NSCLC tissue by immunohistochemistry and found that YAP was expressed in 66.3% (61/92) cases and predominantly presented in the nucleus. The expression of YAP in NSCLC was significantly correlated with p-TNM stage (P = 0.0037) and lymph node metastasis (P = 0.0093). Importantly, YAP expression was associated with short overall survival. Further study in NSCLC cell lines in which YAP was either overexpressed or depleted confirmed that YAP markedly promoted cell proliferation and invasion. These results indicate that YAP plays an important role in NSCLC and might be a useful therapeutic target of NSCLC. (Cancer Sci 2010).

Numerous proinflammatory cytokines, such as TNFalpha and IL-6, which are nuclear factor kappaB (NF-kappaB) target genes, have been shown to promote proliferation in endometriotic cells, and several other genes involved in promoting growth are also NF-kappaB target genes. The aim of this study was to investigate whether the functional insertion/deletion polymorphism (-94 insertion/deletion ATTG) in the promoter of nuclear factor kappaB gene (NFKB1) is associated with susceptibility to endometriosis. Polymerase chain reaction-polyacrylamide gel electrophoresis method was used to genotype the NFKB1 -94 insertion/deletion ATTG polymorphism in 206 women with endometriosis and 365 ethnicity-matched healthy control women. The genotyping method was confirmed by the DNA sequencing analysis. Genotype at the -94 insertion/deletion ATTG polymorphism in the NFKB1 promoter was in Hardy-Weinberg equilibrium in either case or control subjects. The frequency of the ATTG(2)/ATTG(2) genotype and ATTG(2) allele in the endometriosis was significantly higher than that of control subjects (59.7% vs. 37%, odds ratio = 3.069, p < 0.001 for ATTG(2)/ATTG(2) genotype; 75.2% vs. 59.7%, odds ratio = 2.049, p < 0.001 for ATTG(2) allele), indicating that the -94 insertion/deletion ATTG polymorphism in the NFKB1 promoter was associated with endometriosis. This study suggests that the functional -94 insertion/deletion ATTG polymorphism in the promoter of NFKB1 is associated with an increased risk for endometriosis.

Endometrial carcinoma is one of the most common cancers in women. A limited number of endometrial carcinoma cell lines are available for studies of signal transduction pathways and experimental therapeutics in vitro. However, these cell lines have not been comprehensively characterized. In this study, we used genome-wide microarray-based comparative genomic hybridization (aCGH) technology to characterize five of the more commonly used endometrial cancer cell lines. We detected DNA copy-number gains in chromosomal regions 2q, 3p, 3q, 5q, 7p, 17q, and19q in all five cell lines. Other common sites of copy-number gains, which were detected in four of five cell lines, included segments of chromosomes 1, 6, 8, 9, 11, 12, and 16. In all five cell lines, we found DNA copy-number losses in regions 3p, 10p, 10q, 11q, 11p, 14q, 15q, 18p, and 21q. Other common sites of genetic aberrations included segments of chromosomes 1, 2, 4, 5, 6, 16, 20, and 22. The genes involved in the copy-number alterations included the oncogenes PIK3CA (3q26.3), K-ras (12p12.1), R-ras (19q13.3-qter), Raf-1 (3p25), EGFR (7p12), Akt1 (14q32.32), and Akt2 (19q13.1-q13.2). A pathway analysis showed that genes in the PI3K and Wnt pathways are commonly affected. Our characterization of genomic alterations in these five commonly used endometrial cancer cell lines provides valuable genomic information for research that focuses on these key oncogenic pathways in endometrial cancer.

From the anti-TB active fractions of the inner stem bark of Oplopanax horridus, two new heterocyclic nerolidol derivatives, 3,10-epoxy-3,7,11-trimethyldodeca-1,6-dien-11-ol, named neroplomacrol (1), and rel-(3S,6R,7S,10R)-7,10-epoxy-3,7,11-trimethyldodec-1-ene-3,6,11-triol, named neroplofurol (2), were isolated together with oplopandiol (3), falcarindiol (4), and sesamin (5). Extensive spectroscopic analysis revealed that 1 possesses a novel 3,10-oxanonacyclic ring system. Computer-iterated full spin system analysis led to the generation of (1)H NMR fingerprints that will facilitate future dereplication of analogues by providing characteristic spin-spin coupling patterns. The full spin analysis of 5 revealed asymmetric coupling patterns among the chemically equivalent spins, thus confirming the magnetic asymmetry of 5. It was further demonstrated that (1)H NMR fingerprints and MS data enable dereplication of isolates at a submilligram levels including their relative configuration. Countercurrent concentration of the anti-TB activity of the ethnobotanical O. horridus versus the Mycobacterium tuberculosis Erdman strain led to polyynes 3 and 4 as main anti-TB active principles. Their synergistic behavior is linked to a complex fraction containing the new nerolidiol sesquiterpenes, 1 and 2, as phytochemical marker compounds.

An 1,3-oxybenzylglycine based compound 2 (BMS-687453) was discovered to be a potent and selective peroxisome proliferator activated receptor (PPAR) alpha agonist, with an EC(50) of 10 nM for human PPARalpha and approximately 410-fold selectivity vs human PPARgamma in PPAR-GAL4 transactivation assays. Similar potencies and selectivity were also observed in the full length receptor co-transfection assays. Compound 2 has negligible cross-reactivity against a panel of human nuclear hormone receptors including PPARdelta. Compound 2 demonstrated an excellent pharmacological and safety profile in preclinical studies and thus was chosen as a development candidate for the treatment of atherosclerosis and dyslipidemia. The X-ray cocrystal structures of the early lead compound 12 and compound 2 in complex with PPARalpha ligand binding domain (LBD) were determined. The role of the crystal structure of compound 12 with PPARalpha in the development of the SAR that ultimately resulted in the discovery of compound 2 is discussed.

OBJECTIVE: To study the flavonoids and their antioxidant activities of Artemisia annua. METHODS: Isolation and purification were carried out by silica gel and Sephadex LH-20 column chromatographies. Compounds were identified by physicochemical properties and spectral analysis, then their antioxidant activities were evaluated by ORAC assay. RESULTS: Five flavonoids were isolated from the plant. Their structures were identified as 5-hydroxy-3,7,4'-trimethoxyflavone(1), 5-hydroxy-6,7,3',4'-tetramethoxyflavonol(2), blumeatin(3), 5, 4'-dihydroxy-3,7,3'-trimethoxyflavone(4) and quercetin(5), respectively. Compounds 1-5 could slow up the attenuation rate of the fluorenscence induced by AAPH. CONCLUSION: Compounds 1-3 are isolated from the plant for the first time. Compounds 1-5 all possess the potential antioxidant activities.

BACKGROUND:: Velopharyngeal inadequacy (VPI), which has a significant negative impact on speech intelligibility and resonance quality, may be caused by physiological inadequacy. The current study aimed to investigate the maximal velar and pharyngeal motions and levator muscle shortening in the children with repaired cleft palate and different speech outcomes as well as children without cleft palate by using magnetic resonance imaging techniques without general anesthesia. METHODS:: Three groups of sex- and age-matched children were recruited: children with repaired cleft palate and adequate velopharyngeal function condition (VPC), children with repaired cleft palate and VPI, and the normal controls (noncleft). The children were trained to perform sustained /a:/, /i:/, /ts:/, and /m:/, while keeping the head still during magnetic resonance imaging scan. The maximal velar elevation and stretch, pharyngeal medial constriction, velopharyngeal ratio (VP ratio), and levator muscle shortening ratio were measured and compared across the 3 groups. RESULTS:: The VPI group showed the least maximal velar stretch, lowest maximal velar height, smallest maximal pharyngeal constriction, and lowest maximal VP ratio among the 3 groups. The VPI and VPC groups differed significantly in velar and pharyngeal mobility. The effective VP ratio at rest has a strong correlation with that during sustained phonation across the 3 groups. The maximal velar stretch ratio correlates to the maximal pharyngeal constriction ratio strongly in the VPI group only. CONCLUSIONS:: The VPI group had significantly reduced velar and pharyngeal mobility during speech compared with the VPC and noncleft groups. The possible physiological causes of VPI after primary palatal repair were discussed.

Objective: Recent genome-wide association studies have identified multiple novel loci associated with obesity in Europeans. We hypothesized that these genetic variants may be associated with obesity and type 2 diabetes (T2D) in Chinese. Research Design and Methods: We examined 14 associated single-nucleotide polymorphisms at 12 loci (NEGR1, SEC16B, TMEM18, ETV5/DGKG, GNPDA2, LIN7C/BDNF, MTCH2, BCDIN3D/FAIM2, SH2B1, FTO, MC4R, and KCTD15) in 605 healthy adults, 1087 healthy adolescents and 6013 T2D patients from Hong Kong. Results: The European at-risk alleles at five loci including GNPDA2, BCDIN3D/FAIM2, SH2B1, FTO, and KCTD15 were significantly associated with increased body mass index (BMI), waist circumference (4.5 x 10(-8) < P < 0.024), and/or obesity risk (odds ratio 1.14-1.22, 2.0 x 10(-5) < P < 0.002) in our Chinese populations. The former four loci as well as LIN7C/BDNF were also modestly associated with T2D risk (odds ratio 1.09-1.22, 0.008 < P < 0.041), but the associations were lost after adjustment for BMI, suggesting their roles in T2D risk are mediated through modulation of adiposity. Joint effect analyses of the five adiposity loci revealed an increase of about 0.29 kg/m(2) in BMI with each additional copy of at-risk allele (Ptrend = 4.2 x 10(-12)). Conclusions: Our findings support the important contribution of GNPDA2, BCDIN3D/FAIM2, SH2B1, FTO, and KCTD15 in the regulation of adiposity, which in turn affects T2D risk in Chinese.

Quinolone antibacterial agents are widely used in the clinic because of their high antibacterial activity, broad spectra and favorable pharmacokinetics. However, the adverse effects induced by quinolones, such as tendon/articular toxicity, central nervous system toxicity, photo toxicity and dysglycemia, have greatly restricted their therapeutic use. Differentiated embryo-chondrocyte expressed gene 1 (DEC1), an important transcription factor that has a basic helix-loop-helix domain and is ubiquitously expressed in both human embryonic and adult tissues, has a pivotal function in various biological phenomena, including neurogenesis, neuroregulation, chondrogenesis, cell growth, oncogenesis, immune balance and circandian rhythm. Recently, DEC1 has received increasing attention for its role in maintaining the homeostasis of metabolism and energy. Research has shown that DEC1 may play a vital role in metabolic disease. Although the mechanism of the adverse reactions caused by quinolones has not been clarified, the distribution of these serious adverse effects in tissues and organs is consistent with the expression of DEC1 in corresponding normal tissues. In the present paper, we review evidence showing that DEC1 may take part in the adverse effects induced by quinolone antibiotics. The investigation of the molecular details of the toxicity caused by quinolones may help overcome the shortcomings of the antibiotics and reveal new, useful therapeutic functions besides their antimicrobial effect.

In vitro assays identified the Golgi peripheral protein GRASP65 as a Golgi stacking factor that links adjacent Golgi cisternae by forming mitotically regulated trans-oligomers. These conclusions, however, require further confirmation in the cell. In this study, we demonstrated that the first 112 amino acids at the N-terminus (including the first PDZ domain, PDZ1) of the protein are sufficient for oligomerization. Systematic electron microscopic analysis showed that expression of non-regulatable GRASP65 mutants in HeLa cells enhanced Golgi stacking in interphase and inhibited Golgi fragmentation during mitosis. Depletion of GRASP65 by small interference RNA (siRNA) reduced the number of cisternae in the Golgi stacks; this reduction was rescued by expressing exogenous GRASP65. These results provided evidence and a molecular mechanism by which GRASP65 stacks Golgi cisternal membranes. Further experiments revealed that inhibition of mitotic Golgi disassembly by expressing non-regulatable GRASP65 mutants did not affect equal partitioning of the Golgi membranes into the daughter cells. However, it delayed mitotic entry and suppressed cell growth; this effect was diminished by dispersing the Golgi apparatus with Brefeldin A treatment prior to mitosis, suggesting that Golgi disassembly at the onset of mitosis plays a role in cell cycle progression.

Liquid droplet sliding is an approach for the transportation and manipulation of fluids in micro/nanofluidics related biosensor applications. Liquid droplet sliding is also instructive to evaluate the degree of boundary slip. In this study, liquid microdroplet sliding is studied with and without the presence of an electric field using atomic force microscopy on a polystyrene surface spin coated on a doped silicon wafer with a silicon oxide coating. Droplets with different diameters are slid with an atomic force microscope tip, and the sliding forces or lateral friction forces are quantitatively measured. Experimental results show that lateral friction force linearly increases with increasing droplet diameters, which can be explained by a droplet sliding model. When the electric field is applied, in addition to the decreased contact angle, diameters of the liquid droplets will increase. As a result, the electric field increases lateral friction force. To further investigate the impact of applied potential on lateral friction force, voltages from 0 to 80 V are applied to a droplet. Lateral friction force is found to increase with applied potential.