Helicobacter pylori, a gram-negative bacterial pathogen that expresses a strong urease activity, is associated with the development of gastroduodenal disease. Urease B subunit, one of the two structural subunits of urease, was expressed in E. coli BL21 (DE3) strain. The objective of this study was to evaluate the effects of Helicobacter pylori urease B subunit on the immune responses in mice by subcutaneous immunization.
The mice were immunized and boosted with Helicobacter pylori urease B subunit antigen subcutaneously three times with 2-wk intervals between the immunizations and boosters. The mice in the control group were immunized with PBS. The adjuvant group received PBS containing complete/incomplete freund’s adjuvant identical to antigen group without Helicobacter pylori urease B subunit antigen. Four weeks after the final booster, all the mice were sacrificed. Blood was collected on d 0, 14, 28 and 56 before immunization, booster and sacrifice, respectively. Immediately after sacrifice, gastric liquid and spleen were collected for antibody and cytokine analyses.
Urease B subunit increased the concentrations of serum and gastric anti-urease B antigen specific IgG, and the levels of interleukin-4 and interferon-γ in splenocytes of the mice (P < 0.05).
This study demonstrated that recombinant urease B subunit can induce systemic and local immune responses in mice by subcutaneous immunization, which might be used as the effective component of vaccine against Helicobacter pylori.
Antibody; Cytokines; Helicobacter pylori urease B subunit; Mice
Due to language limitations, little is known about the reporting quality of randomized clinical trials (RCTs) on the treatment of coronary heart disease (CHD) with traditional Chinese medicine (TCM) in Chinese Journal of Integrated Traditional and Western Medicine (CJITWM).
In this study, we utilized the CONSORT 2010 statement to understand the reporting quality of RCTs on CHD with TCM from the CJITWM.
The China National Knowledge Infrastructure (CNKI) electronic database was searched for CJITWM RCTs on the treatment of CHD with TCM, published between Janurary 1, 2006 and December 31, 2011. We excluded articles reported as “animal studies,” “topic review,” “diagnostic test,” “editorials,” or “others.” The CONSORT checklist was applied to evaluate the reporting quality of all eligible articles by two independent authors after extensive discussion. Each item was graded as either “yes” or “no” depending on whether the authors had reported it or not.
We identified 21 articles meeting our inclusion criteria. The percentage of 11 of the 37 items was 4.8∼95.2%, 14 of the 37 items were reported in all included articles, while 12 items were not mentioned at all. The average reporting percentage for the “title and abstract” section was 52.4%, for the “introduction” section 100.0%, for the “methods” section 45.4%, for the “results” section 57.1%, for the “discussion” section 79.4%, and for the “other information” section 17.5%.
In general, the reviewed RCTs were not consistent with the CONSORT 2010 statement. Authors should adhere to the CONSORT statement in reporting RCTs; editorial departments may consider the CONSORT statement as a guideline and should instruct authors to write manuscripts, and reviewers to judge them according to CONSORT statutes.
In developed countries, prostate cancer (PCa) is the second most frequently diagnosed type of cancer and the third most common cause of cancer-related mortality in males. Compared with western countries, the morbidity rate of PCa in China is markedly lower, however, it is rising annually. The etiology of PCa is unclear, therefore, to investigate how a disintegrin and metalloprotease 10 (ADAM10) functions in PCa, ADAM10 mRNA and protein levels induced by tumor necrosis factor (TNF)-α were identified using polymerase chain reaction and flow cytometry, respectively. To investigate the mechanism of ADAM10 activity in PCa, specific inhibitors were used, and DNA transfection and RNA interference technology were employed to identify the interaction between ADAM10 and the Fas ligand (L). The results indicated that TNF-α induced ADAM10 expression in a time- and dose-dependent manner through the p38 mitogen-activated protein kinase/necrosis factor-κB signaling pathway. ADAM10 hydrolyzed FasL and contributed to apoptosis resistance of the tumor cells. These observations indicate a promising therapeutic modality for the treatment of apoptosis-resistant PCa, by targeting ADAM10 sheddase activity.
prostate cancer; a disintegrin and metalloproteinase 10; fas ligand; tumor necrosis factor-α; apoptosis
This paper presents the experimental results of cogasification of coal and biomass in an intermittent fluidized bed reactor, aiming to investigate the influences of operation parameters such as gasification temperature (T), steam to biomass mass ratio (SBMR), and biomass to coal mass ratio (BCMR) on hydrogen-rich (H2-rich) gas production. The results show that H2-rich gas free of N2 dilution is produced and the H2 yield is in the range of 18.25~68.13 g/kg. The increases of T, SBMR, and BCMR are all favorable for promoting the H2 production. Higher temperature contributes to higher CO and H2 contents, as well as H2 yield. The BCMR has a weak influence on gas composition, but the yield and content of H2 increase with BCMR, reaching a peak at the BCMR of 4. The H2 content and yield in the product gas increase with SBMR, whilst the content of CO increases first and then decreases correspondingly. At a typical case, the relative linear sensitivity coefficients of H2 production efficiency to T, SBMR, and BCMR were calculated. The results reveal that the order of the influence of the operation parameters on H2 production efficiency is T > SBMR > BCMR.
The title compound, C6HCl6N, lies on a mirror plane, the asymmetric unit conataining a half-molecule. Weak intramolecular C—H⋯Cl contacts are observed.
MicroRNAs (miRNAs) are a distinct class of small RNAs in plants that not only regulate biological processes but also regulate response to environmental stresses. The toxic heavy metal cadmium (Cd) induces expression of several miRNAs in rapeseed (Brassica napus), but it is not known on a genome-wide scale how the expression of miRNAs and their target genes, is regulated by Cd. In this study, four small RNA libraries and four degradome libraries were constructed from Cd-treated and non-Cd-treated roots and shoots of B. napus seedlings. Using high-throughput sequencing, the study identified 84 conserved and non-conserved miRNAs (belonging to 37 miRNA families) from Cd-treated and non-treated B. napus, including 19 miRNA members that were not identified before. Some of the miRNAs were validated by RNA gel blotting. Most of the identified miRNAs were found to be differentially expressed in roots/shoots or regulated by Cd exposure. The study simultaneously identified 802 targets for the 37 (24 conserved and 13 non-conserved) miRNA families, from which there are 200, 537, and 65 targets, belonging to categories I, II, and III, respectively. In category I alone, many novel targets for miRNAs were identified and shown to be involved in plant response to Cd.
Brassica napus; cadmium; degradome; deep sequencing; microRNAs; Brassica napus
In the title compound, C13H15NO4S, there are two independent but conformationally similar molecules in the asymmetric unit, both having an E conformation of the side-chain C=C group. Intramolecular N—H⋯O and O—H⋯O hydrogen-bonding interactions are present in both molecules. In the crystal, one of the molecule types is linked through intermolecular hydroxy–ketone O—H⋯O interactions, forming one-dimensional chains extending along , whereas the other molecule type shows no associations.
Introduction and hypothesis
The aim of the study was to assess the efficacy of low-frequency electrotherapy (LFE) for female patients with early-stage detrusor underactivity (DUA) due to neuromuscular deficiency.
A total of 102 female patients were divided randomly into four groups: LFE-NC (normal compliance), LFE-LC (low compliance), CON (control)-NC and CON-LC. Patients in the LFE-NC and LFE-LC groups received LFE, and those in the CON-NC and CON-LC groups received conservative treatment. Urodynamic evaluation was performed before and after treatment.
After treatment, 82 % of the LFE-NC regained detrusor contractility, whereas only 2 (8 %) of the CON-NC had normal detrusor contraction. None of LFE-LC or CON-LC regained detrusor contractility (p < 0.01). The per cent of LFE-NC who relied on catheterization for bladder emptying decreased by 43 % (p < 0.01). Those in the LFE-LC, CON-NC and CON-LC groups decreased by only 4, 12 or 0 % (p > 0.05).
LFE was more effective for DUA patients with normal compliance; these patients benefited from LFE, but DUA patients with low compliance did not.
Detrusor underactivity; Electrotherapy; Functional stimulation; Low frequency; Lower urinary tract dysfunction; Urodynamics
The title compound, C16H19NO5, which was synthesized from p-methoxycinnamic acid, has intramolecular O—H⋯O and N—H⋯O hydrogen-bonding interactions. In the crystal, molecules are linked by weak C—H⋯O hydrogen bonds and aromatic π–π stacking interactions [minimum ring centroid–centroid separation = 3.790 (1) Å].
To conserve scarce energetic resources during winter, seasonal breeders inhibit reproduction and other nonessential behavioral and physiological processes. Reproductive cessation is initiated in response to declining day lengths, a stimulus represented centrally as a long-duration melatonin signal. The melatonin signal is not decoded by the reproductive axis directly, but by an unidentified neurochemical system upstream of gonadotropin-releasing hormone (GnRH). The dorsomedial nucleus of the hypothalamus (DMH) has been implicated in seasonal changes in reproductive function in Syrian hamsters (Mesocricetus auratus), although the specific-cell phenotype decoding photoperiodic information remains unknown. RFamide-related peptide (RFRP; the mammalian homolog of the gonadotropin-inhibitory hormone (GnIH) gene identified in birds) has emerged as a potent inhibitory regulator of the reproductive axis and, significantly, its expression is localized to cell bodies of the DMH in rodents. In the present study, the authors explored the relationship between RFRP expression, photoperiod exposure, and reproductive condition/hormonal status. In male hamsters that respond to short days with reproductive inhibition, RFRP-ir and mRNA expression are markedly reduced relative to long-day animals. Replacement of testosterone in short-day animals did not affect this response, suggesting that alterations in RFRP expression are not a result of changing sex steroid concentrations. A subset of the hamster population that ignores day length cues and remains reproductively competent in short days (nonresponders) exhibits RFRP-ir expression comparable to long-day hamsters. Analysis of cell body and fiber density suggests a potential interplay between peptide production and release rate in differentially regulating the reproductive axis during early and late stages of reproductive regression. Together, the present findings indicate that photoperiod-induced suppression of reproduction is associated with changes in RFRP and mRNA expression, providing opportunity for further exploration on the role that RFRP plays in this process.
reproduction; gonad; melatonin; seasonal; photoperiod; DMH
To maximize reproductive success, organisms restrict breeding to optimal times of day or year, when internal physiology and external environmental conditions are suitable for parent and offspring survival. To appropriately coordinate reproductive activity, internal and external standing is communicated to the hypothalamo-pituitary-gonadal (HPG) axis via a coordinated balance of stimulatory and inhibitory neurochemical systems. The cumulative balance of these mediators ultimately drives the pattern of gonadotropin-releasing hormone (GnRH) secretion, a neurohormone that stimulates pituitary gonadotropin secretion. Until 2000, a complementary inhibitor of pituitary gonadotropin secretion had not been identified. At this time, Tsutsui and colleagues uncovered a novel, avian hypothalamic peptide capable of inhibiting gonadotropin secretion in cultured quail pituitary cells. We later examined the presence and functional role for the mammalian ortholog of GnIH, RFamide-related peptide (RFRP-3), in mammals, and confirmed a conserved role for this peptide across several rodent species. To date, a similar distribution and functional role for RFRP-3 have been observed across all mammals investigated, including humans. This overview summarizes the role that RFRP-3 plays in mammals and considers the implications and opportunities for further study by those interested in reproductive physiology and the neural control of sexual behavior and motivation.
photoperiod; ovulation; gonad; luteinizing; gonadotropin
To investigate the associations between the different breast cancer subtypes and survival in Chinese women with operable primary breast cancer.
A total of 1538 Chinese women with operable primary breast cancer were analyzed in this study, the median follow-up was 77 months. Estrogen receptor (ER), progesterone receptor (PR), and HER2 status were available for these patients.
Luminal A (ER+ and/or PR+, HER2-) had a favorable disease-free survival (DFS) and overall survival (OS) compared with other subtypes in the entire cohort. Using the luminal A as a reference, among the patients with lymph node positive disease, HER2+ (ER-, PR-, HER2+) had the worst DFS (hazard ratio, HR=1.80, 95% CI 1.11 to 2.91, P=0.017) and luminal B (ER+ and/or PR+, HER2+) had the worst OS (HR=2.27, 95% CI 1.50 to 3.45, P<0.001); among the patients with lymph node negative disease, triple-negative (ER-, PR-, HER2-) had the worst DFS (HR=2.21, 95% CI 1.43 to 3.41, P<0.001), whereas no significant difference in DFS between HER2+ and luminal B or luminal A was observed.
As compared with luminal A, luminal B and HER2+ have the worst survival in patients with lymph node positive disease, but this is not the case in patients with lymph node negative disease; triple-negative subtype has a worse survival in both lymph node positive and lymph node negative patients.
Breast cancer; Subtypes; Disease-free survival; Overall survival
In the title compound, C4H3Cl2NS, the chloromethyl C and 2-position Cl atoms lie close to the mean plane of the thiazole ring [deviations = 0.0568 (2) and 0.0092 (1) Å, respectively]. No classical hydrogen bonds are found in the crystal structure.
An investigation was conducted to identify the distribution of mosquitoes and mosquito-borne arboviruses in the Qinghai-Tibet Plateau, China from July to August in 2007. A total of 8,147 mosquitoes representing six species from three genera (Aedes, Culex, and Anopheles) were collected in three locations (Geermu city, altitude of 2,780 m; Xining city, 2,200 m; Minhe county, 1,700 m). Six virus isolates were obtained including Tahyna virus (TAHV), Liaoning virus, and Culex pipiens pallens Densovirus. A serosurvey showed immunoglobulin G antibodies by immunofluorescence assay (IFA) against TAHV in residents of all three locations. The IFA-positive human samples were confirmed by 90% plaque-reduction neutralization tests (PRNT90) against TAHV with titers ranging from 1:20 to 1:10,240. In addition, TAHV seropositive cows, sheep, and swine were found in these locations. This investigation represents the first isolation of TAHV from Ae. (Och.) detritus and the first evidence of TAHV infection in residents and livestock in the Qinghai-Tibet Plateau.
There are four molecules in the asymmetric unit of the title compound, C16H17N3O4, in which the dihedral angles between the indole ring system and maleimide ring are 4.5 (3), 8.3 (3), 8.4 (2) and 10.4 (2)°. In the crystal, molecules are linked by numerous N—H⋯O and O—H⋯O hydrogen bonds, generating a three-dimensional network.
Within species, color morphs may enhance camouflage, improve communication and/or confer reproductive advantage. However, in the male cichlid Astatotilapia burtoni, body color may also signal a behavioral strategy. A. burtoni live in a lek-like social system in Lake Tanganyika, Africa where bright blue or yellow territorial (T) males (together ~ 10–30% of the population) are reproductively capable and defend territories containing food with a spawning site. In contrast, nonterritorial (NT) males are smaller, cryptically colored, shoal with females and have regressed gonads. Importantly, males switch between these social states depending on their success in aggressive encounters. Yellow and blue morphs were thought to be adaptations to particular habitats, but they co-exist both in nature and in the laboratory. Importantly, individual males can switch colors so we asked whether color influences behavioral and hormonal profiles. When pairing territorial males with opposite colored fish, yellow males became dominant over blue males significantly more frequently. Moreover, yellow T males had significantly higher levels of 11-ketotosterone than blue T males while only blue NT males had higher levels of the stress hormone cortisol compared to the other groups. Thus color differences alone predict dominance status and hormone profiles in T males. Since T males can and do change color, this suggests that A. burtoni may use color as a flexible behavioral strategy.
A 69-year-old woman with angina had a lesion in the left lower lobe on chest film. Angiography revealed coronary artery disease in three vessels. Combined off pump coronary artery bypass grafting (CABG) and left lower lobectomy were performed through median sternotomy. This approach avoids complications due to staged operations and cardiopulmonary bypass (CPB). This report shows that simultaneous off pump CABG and pulmonary operations can be performed safely in patients with coronary artery disease (CAD) associated with lung cancer.
lung cancer; coronary artery disease; lung resection; off-pump coronary artery bypass grafting
Quantitative real-time polymerase chain reactions (qRT-PCR) have become the method of choice for rapid, sensitive, quantitative comparison of RNA transcript abundance. Useful data from this method depend on fitting data to theoretical curves that allow computation of mRNA levels. Calculating accurate mRNA levels requires important parameters such as reaction efficiency and the fractional cycle number at threshold (CT) to be used; however, many algorithms currently in use estimate these important parameters. Here we describe an objective method for quantifying qRT-PCR results using calculations based on the kinetics of individual PCR reactions without the need of the standard curve, independent of any assumptions or subjective judgments which allow direct calculation of efficiency and CT. We use a four-parameter logistic model to fit the raw fluorescence data as a function of PCR cycles to identify the exponential phase of the reaction. Next, we use a three-parameter simple exponent model to fit the exponential phase using an iterative nonlinear regression algorithm. Within the exponential portion of the curve, our technique automatically identifies candidate regression values using the P-value of regression and then uses a weighted average to compute a final efficiency for quantification. For CT determination, we chose the first positive second derivative maximum from the logistic model. This algorithm provides an objective and noise-resistant method for quantification of qRT-PCR results that is independent of the specific equipment used to perform PCR reactions.
quantitative polymerase chain reaction; four-parameter logistic model; three-parameter simple exponent model; noise-resistant algorithm
Circadian rhythms in behavior and physiology are orchestrated by a master biological clock located in the suprachiasmatic nucleus (SCN). Circadian oscillations are a cellular property, with ‘clock’ genes and their protein products forming transcription-translation feedback loops that maintain 24-hour rhythmicity. Although the expression of clock genes is thought to be ubiquitous, the function of local, extra-SCN timing mechanisms remains elusive. We hypothesized that extra-SCN clock genes control local temporal sensitivity to upstream modulatory signals, allowing system-specific processes to be carried out during individual, optimal times of day. To test this possibility, we examined changes in the sensitivity of immortalized GnRH neurons, GT1-7 cells, to timed stimulation by two key neuropeptides thought to trigger ovulation on the afternoon of proestrus, kisspeptin and vasoactive intestinal polypeptide (VIP). We noted a prominent daily rhythm of clock gene expression in this cell line. GT1-7 cells also exhibited daily changes in cellular peptide expression and GnRH secretion in response to kisspeptin and VIP stimulation. These responses occurred without changes in GnRH transcription. These findings are consistent with the notion that GnRH cells are capable of intrinsic circadian cycles that may be fundamental for coordinating daily changes in sensitivity to signals impacting the reproductive axis.
Gonadotropin-releasing hormone; Kisspeptin; Vasoactive intestinal polypeptide; Reproduction
Circadian rhythms in behavior and physiology are orchestrated by a master biological clock located in the suprachiasmatic nucleus (SCN). Circadian oscillations are a cellular property, with ‘clock’ genes and their protein products forming transcription-translation feedback loops that maintain 24-hour rhythmicity. Although the expression of clock genes is thought to be ubiquitous, the function of local, extra-SCN timing mechanisms remains elusive. We hypothesized that extra-SCN clock genes control local temporal sensitivity to upstream modulatory signals, allowing system-specific processes to be carried out during individual, optimal times of day. To test this possibility, we examined changes in the sensitivity of immortalized GnRH neurons, GT1–7 cells, to timed stimulation by two key neuropeptides thought to trigger ovulation on the afternoon of proestrus, kisspeptin and vasoactive intestinal polypeptide (VIP). We noted a prominent daily rhythm of clock gene expression in this cell line. GT1–7 cells also exhibited daily changes in cellular peptide expression and GnRH secretion in response to kisspeptin and VIP stimulation. These responses occurred without changes in GnRH transcription. These findings are consistent with the notion that GnRH cells are capable of intrinsic circadian cycles that may be fundamental for coordinating daily changes in sensitivity to signals impacting the reproductive axis.
Gonadotropin-releasing hormone; Kisspeptin; Vasoactive intestinal polypeptide; Reproduction
Gonadotropin releasing hormone 1 (GnRH1) causes the release of gonadotropins from the pituitary to control reproduction. Here we report that two heterogeneous nuclear ribonucleoproteins (hnRNP-A/B and hnRNP-G) bind to the GnRH-I upstream promoter region in a cichlid fish, Astatotilapia burtoni. We identified these binding proteins using a newly developed homology based method of mass spectrometric peptide mapping. We show that both hnRNP-A/B and hnRNP-G co-localize with GnRH1 in the pre-optic area of the hypothalamus in the brain. We also demonstrated that these ribonucleoproteins exhibit similar binding capacity in vivo, using immortalized mouse GT1-7 cells where overexpression of either hnRNP-A/B or hnRNP-G significantly down-regulate GnRH1 mRNA levels in GT1-7 cells, suggesting that both act as repressors in GnRH1 transcriptional regulation.
Gonadotropin releasing hormone 1 (GnRH1); heterogeneous nuclear ribonucleoprotein A/B (hnRNP-A/B); heterogeneous nuclear ribonucleoprotein G (hnRNP-G); Transcriptional regulation; Mass spectrometric peptide mapping; Hypothalamic-pituitary-gonadal (HPG) axis
In the molecule of the title compound, C9H9NOS, the seven-membered ring has a twist conformation. In the crystal structure, intermolecular N—H⋯O hydrogen bonds link the molecules into centrosymmetric dimers.
Retinal development occurs in mice between embryonic day E11.5 and post-natal day P8 as uncommitted neuroblasts assume retinal cell fates. The genetic pathways regulating retinal development are being identified but little is understood about the global networks that link these pathways together or the complexity of the expressed gene set required to form the retina. At E14.5, the retina contains mostly uncommitted neuroblasts and newly differentiated neurons. Here we report a sequence analysis of an E14.5 retinal cDNA library. To date, we have archived 15 268 ESTs and have annotated 9035, which represent 5288 genes. The fraction of singly occurring ESTs as a function of total EST accrual suggests that the total number of expressed genes in the library could approach 27 000. The 9035 ESTs were categorized by their known or putative functions. Representation of the genes involved in eye development was significantly higher in the retinal clone set compared with the NIA mouse 15K cDNA clone set. Screening with a microarray containing 864 cDNA clones using wild-type and brn-3b (–/–) retinal cDNA probes revealed a potential regulatory linkage between the transcription factor Brn-3b and expression of GAP-43, a protein associated with axon growth. The retinal EST database will be a valuable platform for gene expression profiling and a new source for gene discovery.
The Rhs elements are complex genetic composites widely spread among Escherichia coli isolates. One of their components, a 3.7-kb, GC-rich core, maintains a single open reading frame that extends the full length of the core and then 400 to 600 bp beyond into an AT-rich region. Whereas Rhs cores are homologous, core extensions from different elements are dissimilar. Two new Rhs elements from strains of the ECOR reference collection have been characterized. RhsG (from strain ECOR-11) maps to min 5.3, and RhsH (from strain ECOR-45) maps to min 32.8, where it lies in tandem with RhsE. Comparison of strain K-12 to ECOR-11 indicates that RhsG was once present in but has been largely deleted from an ancestor of K-12. Phylogenetic analysis shows that the cores from eight known elements fall into three subfamilies, RhsA-B-C-F, RhsD-E, and RhsG-H. Cores from different subfamilies diverge 22 to 29%. Analysis of substitutions that distinguish between subfamilies shows that the origin of the ancestral core as well as the process of subfamily separation occurred in a GC-rich background. Furthermore, each subfamily independently passed from the GC-rich background to a less GC-rich background such as E. coli. A new example of core-extension shuffling provides the first example of exchange between cores of different subfamilies. A novel component of RhsE and RhsG, vgr, encodes a large protein distinguished by 18 to 19 repetitions of a Val-Gly dipeptide occurring with a eight-residue periodicity.