Genome mining of actinomycin shunt products from Kitasatospora sp. YINM00002.

Actinomycins are known for their anti-tumor, antibacterial and antiviral activities, and in particular for the ability of actinomycin D as a clinical drug to treat a variety of cancers. In our ongoing work to obtain novel natural products from endophytic actinomycetes derived from traditional Chinese herbs, we identified the potential to produce actinomycins in YINM00002, a Kitasatospora strain derived from Polygonatum kingianum. According to genome mining, we isolated actinomycins D and V (1 and 2) and small amounts of 4-methyl-3-hydroxyanthranilic acid (4-MHA) derivates (3 and 4) from strain fermentation broth. The presence of actinrhater A (3) and actinrhater B (4) reveals a mysterious shunt pathway in the early stages of actinomycin D biosynthesis. Our study provides a fresh perspective for further discovery and modification of novel actinomycins.

High glucose-increased miR-200c contributes to cellular senescence and DNA damage in neural stem cells.

BACKGROUND: Maternal diabetes increases the risk for neural tube defects (NTDs). It is unclear if miRNAs, senescence, and DNA damage are involved in this process. In this study, we used neural stem cells as an in vitro proxy of embryonic neuroepithelium to investigate whether high glucose triggers neural stem cell senescence and DNA damage by upregulating miR-200c, which may be responsible for NTDs. METHODS: C17.2 neural stem cells were cultured with normal glucose (5 mM) or high glucose (≥16.7 mM) at different doses and time points for detecting miR-200c levels, markers of senescence and DNA damage. Neural stem cells were exposed to antioxidant SOD1 mimetic Tempol and high glucose for 48 h to test roles of oxidative stress on the miR-200c, senescence, and DNA damage levels. An miR-200c mimic and an inhibitor were transfected into neural stem cells to increase or decrease miR-200c activities. RESULTS: High glucose upregulated miR-200c in neural stem cells. A time course study of the effect of high glucose revealed that miR-200c initially increased at 12 h and reached its zenith at 18 h. Tempol reduced miR-200c levels caused by high glucose. High glucose induced markers of senescence and DNA damage in neural stem cells. Tempol abolished high glucose-induced markers of senescence and DNA damage. The miR-200c inhibitor suppressed high glucose-induced markers of senescence and DNA damage. Treatment with miR-200c mimic imitates high glucose-induced markers of senescence and DNA damage. CONCLUSIONS: We show that high glucose increases miR-200c, which contributes to cellular senescence and DNA damage in neural stem cells and provides a potential pathway for maternal diabetes-induced neural tube defects.

Long-term metal fume exposure assessment of workers in a shipbuilding factory.

This study aims to assess the metal fume exposure of welders and to determine exposure rates for similar exposure groups in a shipyard through the use of Near-field/Far-field (NF/FF) mathematical model and Bayesian decision analysis (BDA) technique. Emission rates of various metal fumes (i.e., total chromium (Cr), iron (Fe), lead (Pb), manganese (Mn), and nickel (Ni)) were experimentally determined for the gas metal arc welding and flux cored arc welding processes, which are commonly used in shipyards. Then the NF/FF field model which used the emission rates were further validated by welding simulation experiment, and together with long-term operation condition data obtained from the investigated shipyard, the predicted long-term exposure concentrations of workers was established and used as the prior distribution in the BDA. Along with the field monitoring metal fume concentrations which served as the likelihood distribution, the posterior decision distributions in the BDA were determined and used to assess workers' long-term metal exposures. Results show that the predicted exposure concentrations (Cp) and the field worker's exposure concentrations (Cm) were statistically correlated, and the high R2 (= 0.81-0.94) indicates that the proposed surrogate predicting method by the NF and FF model was adequate for predicting metal fume concentrations. The consistency in both prior and likelihood distributions suggests the resultant posterior would be more feasible to assess workers' long-term exposures. Welders' Fe, Mn and Pb exposures were found to exceed their corresponding action levels with a high probability (= 54%), indicating preventive measures should be taken immediately. The proposed approach provides a universal solution for conducting exposure assessment with usual limited number of personal exposure data.

Upregulation of microRNA-129-5p inhibits cell invasion, migration and tumor angiogenesis by inhibiting ZIC2 via downregulation of the Hedgehog signaling pathway in cervical cancer.

Recently, some studies have placed additional research focus on microRNAs (miRNAs) in a bid to discover novel therapeutic approaches for cervical cancer (CC), which is one of the most common female reproductive tract malignancies with high rates of morbidity and mortality. Hence, the aim of the present study was to evaluate the ability of miR-129-5p to influence cell angiogenesis, invasion and migration by targeting ZIC2 through the Hedgehog signaling pathway in CC. Both CC and adjacent normal tissues were extracted from 87 eligible participating patients with CC. Measurements of the levels of miR-129-5p, mRNA and protein levels of ZIC2, sonic Hedgehog (Shh), Gli1, and Gli2 and levels of CXCL1, VEGF and Ang2 were determined accordingly. An angiogenesis assay was performed to evaluate cell angiogenesis in vitro, while a scratch test and transwell assay were adopted for cell invasion and migration determination. Lastly, tumor formation within nude mice was performed in order to analyze angiogenesis and tumor growth among the nude mice in vivo. The findings revealed that upregulation of miR-129-5p resulted in the decrease in the mRNA and protein levels of ZIC2, Shh, Gli1, Gli2, as well as reduced levels of CXCL1, VEGF and Ang2. Moreover, up-regulation of miR-129-5p was determined to inhibit CC cell angiogenesis ability in vitro, in addition to the processes of cell migration, and invasion. Finally, up-regulation of miR-129-5p was observed to inhibit the tumor growth and angiogenesis ability of nude mice in vivo. The results of the present study provided evidence suggesting that overexpressed miR-129-5p prevents angiogenesis and inhibits cell migration and invasion by means of negatively targeting ZIC2 through suppression of the Hedgehog signaling pathway in CC. Thus, highlighting the promise of miR-129-5p as a novel target for treating CC is promising.

Strategies for imputing missing covariates in accelerated failure time models.

Missing covariates often occur in biomedical studies with survival outcomes. Multiple imputation via chained equations (MICE) is a semi-parametric and flexible approach that imputes multivariate data by a series of conditional models, one for each incomplete variable. When applying MICE, practitioners tend to specify the conditional models in a simple fashion largely dictated by the software, which could lead to suboptimal results. Practical guidelines for specifying appropriate conditional models in MICE are lacking. Motivated by a study of time to hip fractures in the Women's Health Initiative Observational Study using accelerated failure time models, we propose and experiment with some rationales leading to appropriate MICE specifications. This strategy starts with specifying a joint model for the variables involved. We first derive the conditional distribution of each variable under the joint model, then approximate these conditional distributions to the extent which can be characterized by commonly used regression models. We propose to fit separate models to impute incomplete variables by the failure status, which is key to generating appropriate MICE specifications for survival outcomes. The proposed strategy can be conveniently implemented with all available imputation software that uses fully conditional specifications. Our simulation results show that some commonly used simple MICE specifications can produce suboptimal results, while those based on the proposed strategy appear to perform well and be robust toward model misspecifications. Hence, we warn against a mechanical use of MICE and suggest careful modeling of the conditional distributions of variables to ensure proper performance.

[Levels of T-Lymphocyte Subsets, IL-17, IL-35 and IFN-γ in Peripheral Blood and Their Clinical Significance in Patients with Multiple Myeloma].

OBJECTIVE: To explore the levels of T-lymphocyte subsets and IL-17, IL-35, IFN-γ in peripheral blood of patients with multiple myeloma(MM) and their clinical significance. METHODS: A total of 86 MM patients in our hospital from January 2014 to January 2017 were enrolled in MM group and 30 healthy persons were enrolled in control group, the CD4+/CD8+ T cells ratio, CD4+CD25high/+ CD127low/- Treg level in peripheral blood were detected by flow cytometer. The levels of IL-17, IL-35 and IFN-γ in peripheral serum were detected by ELISA, and the differences of detected indicators between different groups were compared. RESULTS: Compared with control group, the proportion of CD4+ T cells and CD4+/CD8+ T cells ratio decreased, the proportion of CD8+ T cells and Treg increased in MM group. The differences of T lymphocyte subsets level between group III stage of MM and control group were statistically significant (P<0.05). With enhancing of clinical stages, Treg level showed a increasing trend, especially in III stage (P<0.05), the serum level of IL-17 as followed in turn: III stage>II stage>I stage>control, the serum level of IL-35 and IFN-γ as followed in turn: control>I stage>II stage>III stage (P<0.05). In terms of disease status, the propurtion of Treg cells as fllowed in turn: disease progression stage>stable stage>control (P<0.05), the serum level of IL-17 as followed in turn also: disease progression stage>stable stage (P<0.05), while the serum level of IL-35 and IFN-γ as followed in turn: control>disease table stage>progression stage (P<0.05). CONCLUSION: The abnomal level of T-lymphocyte subsets, Treg, IL-17, IL-35 and IFN-γ are related with progression and prognosis of MM patients.

Research on differentially expressed genes related to substance and energy metabolism between healthy volunteers and splenasthenic syndrome patients with chronic superficial gastritis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To analyze the metabolic states of the lipids, protein, carbohydrate, and nucleic acid for chronic superficial gastritis patients of splenasthenic syndrome (SS), and to explore the pathogenesis mechanism of SS based on substance and energy metabolisms.</p><p><b>METHODS</b>During June 2004 to March 2005, recruited were four chronic superficial gastritis patients of SS who visited at the First Hospital of Guangzhou University of Chinese Medicine and Guangdong Provincial Hospital of Traditional Chinese Medicine. Four healthy volunteers were recruited from Guangzhou University of Chinese Medicine. Their gastric mucosa was extracted to perform experiments of DNA microarray. The dual-channel DNA microarray data were mined and bioinformatics analyzed by BRB ArrayTools and IPA software.</p><p><b>RESULTS</b>Fifteen genes were involved in substance and energy metabolisms in 20 differentially expressed genes, accounting for 75%.Among these genes, one gene was up-regulated, 14 genes down-regulated, and 11 genes were enzyme gene. Differentially expressed genes related to lipid metabolism included ACAA2 and CYP20A1, manifested as fatty acid catabolism and cholesterol transformation. Genes related to protein metabolism included ALDH9A1, ASL, ASS1, PCY-OX1L, RPS28, UBE2D2, UBXN1, B3GNT1, GCNT1, and PPP1R3C, manifested as decreased amino acid metabolism that may affect the biologic processes such as autonomic nerve, urea cycle, etc., reduced protein synthesis, increased ubiquitination of fault fold proteins, and decreased post-translated modification of glycosylation and dephosphorylation. Genes related to carbohydrate metabolism included PPP1R3C, B3GNT1, and GCNT1, manifested as decreased glycogen and glycan syntheses. Genes related to nucleic acid metabolism included RMI1, SMARCD3, and PARP1, manifested as degraded DNA duplication and transcription, and increased DNA damage repair.</p><p><b>CONCLUSIONS</b>The metabolisms of the lipids, protein, carbohydrate, and nucleic acid in chronic superficial gastritis patients of SS obviously decreased, manifested mainly as down-regulated enzyme gene expression. We inferred that these might be one of the vital pathogenesis mechanisms for nutrition dysmetabolism of SS.</p>

[Study on gene differential expressions of substance and energy metabolism in chronic superficial gastritis patients of Pi deficiency syndrome and of pi-wei hygropyrexia syndrome].

OBJECTIVE: To analyze the metabolic levels of energy and substance in chronic superficial gastritis (CSG) patients of Pi deficiency syndrome (PDS) and of Pi-Wei hygropyrexia syndrome (PWHS), including lipid, protein, nucleic acid, carbohydrate, trace element, and energy metabolism, and to study the pathogenesis mechanism of PDS from substance and energy metabolisms. METHODS: Recruited were 8 CSG patients who visited at First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine and Guangdong Provincial Hospital of Traditional Chinese Medicine from June 2004 to March 2005, including 4 patients of PDS and 4 of PWHS. Their gastric mucosae were used for experiments of DNA microarray. The dual-channel DNA microarray data were bioinformatically analyzed by BRB ArrayTools and IPA Software. RESULTS: Obtained were fifty-six differentially expressed genes involved in substance and energy metabolisms with the expression fold more than 2, including 11 genes up-regulated and 45 genes down-regulated. Of them, genes correlated to lipid metabolism included CRLS1, LRP11, FUT9, GPCPD1, PIGL, SULT1A4, B3GNT1, ST8SIA4, and ACADVL, mainly involved in the metabolic processes of fatty acid, cholesterol, phospholipids, and glycolipid. Genes correlated to protein metabolism included ASRGL1, AARSD1, EBNA1BP2, PUM2, MRPL52, C120RF65, PSMB8, PSME2, UBA7, RNF11, FBXO44, ZFYVE26, CHMP2A, SSR4, SNX4, RAB3B, RABL2A, GOLGA2, KDELR1, PHPT1, ACPP, PTPRF, CRKL, HDAC7, ADPRHL2, B3GNT1, ST8SIA4, DDOST, and FUT9, mainly involved in the biosynthesis processes of protein, ubiquitination, targeted transport and post-translation modification. Genes correlated to nucleic acid metabolism included DFFB, FLJ35220, TOP2A, SF3A3, CREB3, CRTC2, NR1D2, MED6, GTF2IRD1, C1ORF83, ZNF773, and ZMYND11, mainly involved in DNA replication and repair, transcription regulation. Genes correlated to carbohydrate metabolism included AGL, B3GNT1, FUT9, ST8SIA4, SULT1A4, DDOST, and PIGL, mainly involved in glucogen degradation and glycoconjugate biosynthesis. Genes correlated to trace element metabolism included COMMD1, SLC39A6, FTL, CHRFAM7A, SCGN, and S100A6, mainly involved in ion metabolisms of copper, zinc, ferri, and calcium. Genes correlated to energy metabolism included AK3 and COX7B, mainly involved in mitochondria structure and oxidative phosphorylation processes. CONCLUSION: The metabolic levels of energy and substance including lipid, protein, nucleic acid, carbohydrate, and trace element were obviously reduced in patients of PDS, which might be an important pathogenesis mechanism for its occurrence.

Gene expression profiling in gastric mucosa from Helicobacter pylori-infected and uninfected patients undergoing chronic superficial gastritis.

Helicobacter pylori infection reprograms host gene expression and influences various cellular processes, which have been investigated by cDNA microarray using in vitro culture cells and in vivo gastric biopsies from patients of the Chronic Abdominal Complaint. To further explore the effects of H. pylori infection on host gene expression, we have collected the gastric antral mucosa samples from 6 untreated patients with gastroscopic and pathologic confirmation of chronic superficial gastritis. Among them three patients were infected by H. pylori and the other three patients were not. These samples were analyzed by a microarray chip which contains 14,112 cloned cDNAs, and microarray data were analyzed via BRB ArrayTools software and Ingenuity Pathways Analysis (IPA) website. The results showed 34 genes of 38 differentially expressed genes regulated by H. pylori infection had been annotated. The annotated genes were involved in protein metabolism, inflammatory and immunological reaction, signal transduction, gene transcription, trace element metabolism, and so on. The 82% of these genes (28/34) were categorized in three molecular interaction networks involved in gene expression, cancer progress, antigen presentation and inflammatory response. The expression data of the array hybridization was confirmed by quantitative real-time PCR assays. Taken together, these data indicated that H. pylori infection could alter cellular gene expression processes, escape host defense mechanism, increase inflammatory and immune responses, activate NF-κB and Wnt/ß-catenin signaling pathway, disturb metal ion homeostasis, and induce carcinogenesis. All of these might help to explain H. pylori pathogenic mechanism and the gastroduodenal pathogenesis induced by H. pylori infection.

Size distributions and exposure concentrations of nanoparticles associated with the emissions of oil mists from fastener manufacturing processes.

The aims of the present study were set out to measure size distributions and estimate workers' exposure concentrations of oil mist nanoparticles in three selected workplaces of the forming, threading, and heat treating areas in a fastener manufacturing plant by using a modified electrical aerosol detector (MEAD). The results were further compared with those simultaneously obtained from a nanoparticle surface area monitor (NSAM) and a scanning mobility particle sizer (SMPS) for the validation purpose. Results show that oil mist nanoparticles in the three selected process areas were formed mainly through the evaporation and condensation processes. The measured size distributions of nanoparticles were consistently in the form of uni-modal. The estimated fraction of nanoparticles deposited on the alveolar (AV) region was consistently much higher than that on the head airway (HD) and tracheobronchial (TB) regions in both number and surface area concentration bases. However, a significant difference was found in the estimated fraction of nanoparticles deposited on each individual region while different exposure metrics were used. Comparable results were found between results obtained from both NSAM and MEAD. After normalization, no significant difference can be found between the results obtained from SMPS and MEAD. It is concluded that the obtained MEAD results are suitable for assessing oil mist nanoparticle exposures.

A comparison of multiple imputation and fully augmented weighted estimators for Cox regression with missing covariates.

Several approaches exist for handling missing covariates in the Cox proportional hazards model. The multiple imputation (MI) is relatively easy to implement with various software available and results in consistent estimates if the imputation model is correct. On the other hand, the fully augmented weighted estimators (FAWEs) recover a substantial proportion of the efficiency and have the doubly robust property. In this paper, we compare the FAWEs and the MI through a comprehensive simulation study. For the MI, we consider the multiple imputation by chained equation and focus on two imputation methods: Bayesian linear regression imputation and predictive mean matching. Simulation results show that the imputation methods can be rather sensitive to model misspecification and may have large bias when the censoring time depends on the missing covariates. In contrast, the FAWEs allow the censoring time to depend on the missing covariates and are remarkably robust as long as getting either the conditional expectations or the selection probability correct due to the doubly robust property. The comparison suggests that the FAWEs show the potential for being a competitive and attractive tool for tackling the analysis of survival data with missing covariates.

Using a modified electrical aerosol detector to predict nanoparticle exposures to different regions of the respiratory tract for workers in a carbon black manufacturing industry.

The present study was set out to characterize nanoparticle exposures in three selected workplaces of the packaging, warehouse, and pelletizing in a carbon black manufacturing plant using a newly developed modified electrical aerosol detector (MEAD). For confirmation purposes, the MEAD results were compared with those simultaneously obtained from a nanoparticle surface area monitor (NSAM) and a scanning mobility particle sizer (SMPS). We found that workplace background nanoparticle concentrations were mainly coming from the outdoor environment. Size distributions of nanoparticles for the three selected process areas during the work hours were consistently in the form of bimodel. Unlike nanoparticles of the second mode (simply contributed by the process emissions), particles of the first mode could be also contributed by the forklift exhaust or fugitive emissions of heaters. The percents of nanoparticles deposited on the alveolar (A) region were much higher than the other two regions of the head airway (H), tracheobronchial (TB) for all selected workplaces in both number and surface area concentrations. However, significant differences were found in percents of nanoparticles deposited on each of the three regions while different exposure metrics were adopted. Both NSAM and MEAD obtained quite comparable results. No significant difference can be found between the results obtained from SMPS and MEAD after being normalized. Considering the MEAD is less expensive, less bulky, and easy to use, our results further support the suitability of using MEAD in the field for nanoparticle exposure assessments.

Bioinformatics research on chronic superficial gastritis of Pi-deficiency syndrome by gene arrays.

OBJECTIVE: To determine the bioinformatical characteristics of differential gene expression in patients with chronic superficial gastritis (CSG) with the Pi-deficiency syndrome (PDS) and those of the non-Pi-deficiency syndrome (non-PDS), i.e. patients of CSG with Pi-Wei dampnese-heat syndrome and healthy persons. METHODS: With the BRB-Array Tools software package, original data collection and bioinformatic: analysis of gene arrays were conducted in 6 CSG patients of PDS (CSG-PDS), 6 CSG patients of non-PDS (CSG-nPDS), and 6 healthy volunteers (Normal). RESULTS: Compared with non-PDS, the gene expressions: in PDS with regards to protein synthesis, energy metabolism, immune reaction and ionic transport tended to be down-regulated, while those concerning secretion, cytoskeleton and ubiquitinization were up-regulated dominantly. CONCLUSIONS: The two kinds of samples, CSG-PDS/Normal and CSG-PDS/CSG-nPDS, have their respective gene expression profiles with different characteristics. Gene expression profile has certain referential significance in syndrome classification.

Assessing inhalatory and dermal exposures and their resultant health-risks for workers exposed to polycyclic aromatic hydrocarbons (PAHs) contained in oil mists in a fastener manufacturing industry.

This study first assessed workers' inhalatory and dermal exposures to polycyclic aromatic hydrocarbons (PAHs) contained in oil mists. Then, their resultant lung cancer and skin cancer risks were estimated. Finally, control strategies were initiated from the health-risk management aspect. All threading workers in a fastener manufacturing plant were included. 16 inhalatory and 88 dermal PAH exposure samples were collected. Results show that the inhalatory gas phase total PAH exposure level (8.60 x 10(4) ng/m3) was much higher than that of particle phase (2.30 x 10(3) ng/m3). Workers' mean inhalatory exposure level (8.83 x 10(4) ng/m3) was lower, but its corresponding 1-sided upper 95% confidence level (UCL1,95% = 1.02 x 10(5) ng/m3) was higher than the time-weighted average permissible exposure level (PEL-TWA) regulated in Taiwan for PAHs (1.00 x 10(5) ng/m3). The mean whole body total PAHs dermal exposure levels was 5.44 x 10(6) ng/day and the top five exposed surface areas were lower arm, hand, upper arm, neck, and head/front. The estimated lifetime skin cancer risk (9.72 x 10(-3)) was lower than that of lung cancer risk (1.64 x 10(-2)), but both were higher than the significant risk level (10(-3)) defined by the US Supreme Court in 1980. The installation of a local exhaust ventilation system at the threading machine should be considered as the first priority measurement because both lung and skin cancer risks can be reduced simultaneously. If the personal protection equipment would be adopted in the future, both respiratory protection equipment and protective clothing should be used simultaneously.

[A survey on reproductive health related sexual behavior among middle school students in Luoyang city].

OBJECTIVE: To explore the appropriate ways and contents of reproductive health education for middle school students and to understand reproductive health related sexual behavior and influencing factors among middle school students. METHODS: Reproductive health related sexual behavior was evaluated among junior and senior middle school students in Luoyang by cluster sampling. The statistical software of SAS 8.1 was adopted for data analyses. Sexual behavior and influencing factors were analyzed by logistic regression. RESULTS: Critical sexual behaviors were found significantly higher in senior students, including masturbation, sexual fantasy and sexual intercourse than that in junior students (P < 0.05), and boys had higher prevalence than girls (P < 0.05). Results from multivariate logistic regression model analyses indicated that incidence rate of sexual behavior among those who ever having had experiences was higher than those who were inexperienced (OR = 2.62, 95%CI: 1.21 - 5.66). Incidence rate of sexual behavior was related to access of reproductive health and STD/AIDS knowledge (OR = 3.09, 95%CI: 1.43 - 6.51). In addition, incidence rate of sexual behavior was related to attitude and relation of amour between boys and girls (OR = 2.24, 95%CI: 1.32 - 3.75). CONCLUSION: Awareness on reproductive health knowledge among middle school students was not enough. Marginal sexual behaviors as masturbation and sexual fantasy had not been correctly and openly discussed to avoid inappropriate sexual activities.