Full length transcriptomes analysis of cold-resistance of Apis cerana in Changbai Mountain during overwintering period.

Apis cerana in Changbai Mountain is an ecological type of Apis cerana, which is an excellent breeding material with cold-resistant developed by long-term natural selection under the ecological conditions. However, the physiological and molecular mechanisms of Changbai Mountain population under cold stress are still unclear. In this study, the Nanopore sequencing was carried out for the transcriptome of Apis cerana in Changbai Mountain in the coldest period of overwintering, which will provide a reference to the cold-resistant mechanism. We determined 5,941 complete ORF sequences, 1,193 lncRNAs, 619 TFs, 10,866 SSRs and functional annotations of 11,599 new transcripts. Our results showed that the myosin family and the C2H2 zinc finger protein transcription factor family possibly have significant impacts on the response mechanism of cold stress during overwintering. In addition, the cold environment alters genes expression profiles in honeybees via different AS and APA mechanisms. These altered genes in Hippo, Foxo, and MARK pathways help them counter the stress of cold in overwinter period. Our results might provide clues about the response of eastern honeybees to extreme cold, and reflect the possible genetic basis of physiological changes.

Transcriptome analysis of sika deer in China.

Sika deer is of great commercial value because their antlers are used in tonics and alternative medicine and their meat is healthy and delicious. The goal of this study was to generate transcript sequences from sika deer for functional genomic analyses and to identify the transcripts that demonstrate tissue-specific, age-dependent differential expression patterns. These sequences could enhance our understanding of the molecular mechanisms underlying sika deer growth and development. In the present study, we performed de novo transcriptome assembly and profiling analysis across ten tissue types and four developmental stages (juvenile, adolescent, adult, and aged) of sika deer, using Illumina paired-end tag (PET) sequencing technology. A total of 1,752,253 contigs with an average length of 799 bp were generated, from which 1,348,618 unigenes with an average length of 590 bp were defined. Approximately 33.2 % of these (447,931 unigenes) were then annotated in public protein databases. Many sika deer tissue-specific, age-dependent unigenes were identified. The testes have the largest number of tissue-enriched unigenes, and some of them were prone to develop new functions for other tissues. Additionally, our transcriptome revealed that the juvenile-adolescent transition was the most complex and important stage of the sika deer life cycle. The present work represents the first multiple tissue transcriptome analysis of sika deer across four developmental stages. The generated data not only provide a functional genomics resource for future biological research on sika deer but also guide the selection and manipulation of genes controlling growth and development.

Prevalence, risk factors and molecular characterization of Enterocytozoon bieneusi in raccoon dogs (Nyctereutes procyonoides) in five provinces of Northern China.

The disease microsporidiosis is found worldwide and is mainly caused by Enterocytozoon bieneusi. E. bieneusi can infect a wide range of hosts; however, information regarding the prevalence and genotyping of E. bieneusi infection in raccoon dogs (Nyctereutes procyonoides) is limited. Therefore, in 2015, we examined 305 faecel samples from 80 farmed raccoon dogs in Jilin Province, from 54 in Hebei Province, from 72 in Liaoning Province, from 29 in Shandong Province, and from 40 in Heilongjiang Province. The overall prevalence of E. bieneusi infection in farmed raccoon dogs was 22.30%. Logistic regression analysis suggests that age, gender and region of raccoon dogs were highly related to the prevalence of E. bieneusi infection. Moreover, six E. bieneusi internal transcribed spacer (ITS) region sequences, including four known genotypes, namely D, CHN-DC1, NCF2, and CHN-F1, and two novel genotypes (NCR1 and NCR2), were identified in the present study. The present study firstly indicated the existence of E. bieneusi genotypes NCF2, NCR1, NCR2and CHN-F1 in infected raccoon dogs in Northern China. Integrated control strategies should be implemented to limit E. bieneusi infection in farmed raccoon dogs, and to prevent transmission of this disease to other animals and humans.

[Eenie, Meenie, Miney, Moe, who is responsible for the antibody-dependent enhancement of Aleutian mink disease parvovirus infection?].

Aleutian mink disease parvovirus (AMDV) causes a persistent infection associated with immune complex disease, hypergammaglobulinemia, and high levels of antiviral antibodies. Despite the presence of an antibody, the virus is not cleared in vivo. Pre-existing antibodies may enhance viral infections, by Fc-receptor-mediated antibody-dependent enhancement (ADE), but the mechanism that underlies ADE has not been fully defined. Three models have been proposed, including: (1) interactions between antibody and FcR, complement C3 fragment and CR, or between C1q and C1qR, which promotes viral attachment to cells; (2) suppression of IFN-gamma-mediated host-cell antiviral gene expression by the upregulation of negative regulators of pathogen pattern recognition; and (3) the promotion of early IL-10 secretion. In addition, the role of cytokine IL-6 in ADE mediated disease development is discussed, to facilitate a better understanding of the pathogenesis of AMDV infection, as well as give insights into rational vaccine design approaches.

PP2A-AMPKα-HSF1 axis regulates the metal-inducible expression of HSPs and ROS clearance.

Metals such as cadmium and arsenic are ubiquitous toxicants that cause a variety of adverse health effects. Heat shock proteins (HSPs) response to metal-induced stress and protect cells from further damage. However, the intracellular signalling pathways responsible for activation of HSPs expression are not fully understood. Here, we demonstrate that protein phosphatase 2A (PP2A) regulates expression of HSP70 and HSP27 via dephosphorylation of an AMP-activated protein kinase α subunit (AMPKα) at Thr172. Dephosphorylated AMPKα phosphorylates heat shock factor 1 (HSF1) at Ser303, leading to significant transcriptional suppression of HSP70 and HSP27 in CdCl2- or NaAsO2-treated cells. Suppression of PP2A regulatory B56δ subunit resulted in the sustained phosphorylation of AMPKα upon CdCl2 treatment, subsequent reduction in expression of HSP70 and HSP27, and thereby dramatic reduction of reactive oxygen species (ROS) clearance. We further revealed that PP2A B56δ physically interacted with AMPKα, providing evidence that PP2A B56δ-AMPKα-HSF1 signalling pathway participated in regulating the inducible expression of HSPs and ROS clearance. Taken together, we identified a novel PP2A-dependent signalling pathway involved in regulation of HSPs expression in response to metal stress.

[The analysis of the active follow-up study of registered cancer patients between 2002 and 2005 in urban areas of Beijing].

OBJECTIVE: To evaluate the results of the active follow-up among registered cancer patients in 2002 - 2005 in urban areas of Beijing. METHODS: A number of 63 997 cancer patients diagnosed during 2002 - 2005 were selected from the surveillance database of Beijing Office for Cancer Prevention and Control. By matching the identity information of the patients with the death surveillance database built by the vital statistic department in Beijing, 29 223 patients were confirmed to be alive.1149 cases were removed from the study due to lack of exact key variables, such as address and telephone numbers. 28 074 patients were, at last, included in the active follow-up study. The investigators and the inspectors, who accepted standard training program, investigated each patient's status of census register and survival condition by phone calling, household interview and visits at local police station or residential committee. The loss ratio of follow-up and the constituent ratio of the withdrawal reasons were calculated. RESULTS: Among the 28 074 patients selected in active follow-up, 21 696 patients were followed successfully; 1453 of whom didn't have the census register of Beijing, which accounted for 6.70%. Out of the other 20 243 Beijing residents, 4715 patients (23.29%) were already dead and 84.22% (3971/4715) of them replenished the failure to report by passive follow-up. Among all the 4715 dead cases, 4405 (93.43%) patients were died from cancer. The follow-up study helped to replenish the vital statistics in different districts, the ratio ranged from 4.87% and 8.85%. 6378 patients were withdrawn from the study. The loss ratio was 22.72% (6378/28 074), and the total loss ratio was 12.03% ((6378 + 1149)/(63 997 - 1453)). Of these withdrawal cases, 3041 (47.68%) were lost to follow-up in that the investigators can't find the patients or the relatives of the patients according to the registered phone number or address information. The other reasons included: the patients removed to other areas (1199 cases, 18.80%), the patients and their family members were temporarily not at home (127 cases, 1.99%), the patients and their family members rejected to answer the interview (292 cases, 4.58%), and other reasons (1719 cases, 26.95%). CONCLUSION: The method of active follow-up towards registered cancer patients can replenish the missing information which could not be collected from passive follow-up procedure; and therefore effectively improve the quality of data in cancer registration.

[A genome-wide screen for promoter-specific sites of differential DNA methylation during human cell malignant transformation in vitro].

OBJECTIVE: To explore potential epigenetic biomarkers for toxic effects, tumor-related chemical prevention and biological monitor by a genome-wide screening for differential DNA methylation during human cell malignant transformation in vitro. METHODS: The two in vitro cell transformation models included B(a)P-induced human bronchial epithelial cell introduced by H-Ras (HBER) cell transformation and simian vacuolating virus 40 small T antigen induced (SV40 ST-induced) HBER cell transformation. Methylated genes were collected by methylated DNA immunoprecipitation and whole genome amplification (MeDIP-WGA) at three time points during cell transformation which represented different transformation stage. Then, CpG island microarray was used to screen differentially methylated genes. The mRNA levels of hypermethylated genes were also observed by RT-PCR. RESULTS: The CpG island microarray showed that the number of hypermethylated genes in HBER, HBERNT, HBERT cells were 733, 661 and 738 respectively.83 genes were hypermethylated in pre-transformed cell and transformed cell. Moreover, 25 of 83 genes were also hypermethylated in SV40 ST-transformed cell (HBERST). We further confirmed that the mRNA expression of six of these 25 genes, namely family with sequence similarity 178, member A (FAM178A), retinoic acid receptor responder (tazarotene induced) (RARRES1), ubiquitin specific peptidase 28 (USP28), Scm-like with four mbt domains 2 (SFMBT2), family with sequence similarity 59, member A (FAM59A) and nuclear receptor subfamily 4, group A, member 3 (NR4A3) were suppressed during B(a)P-induced transformation. CONCLUSION: The abnormal hypermethylation of specific genes was a common event in the two kinds of human cell transformation models, which shed light on the study for chemical exposure monitor and tumor-related epigenetic biomarkers.

[Incidence trends and pathological characteristics of lung cancer in urban Beijing during period of 1998 - 2007].

OBJECTIVE: To describe the incidence trends and pathological characteristics of lung cancer in urban Beijing, China. METHODS: A total of 32 845 medical records of the residents diagnosed as lung cancer in urban Beijing from 1998 to 2007 were retrieved through the cancer registry system of Beijing Cancer Registry. Crude incidence rate, age-specific incidence rate, adjusted incidence rate by world standardized population, annual percentage change (APC) and histological categorized incidence rate by world standardized population were calculated in order to compare the differences of the incidence trends in different time periods, or among different gender and age groups. RESULTS: A total of 32 845 newly diagnosed lung cancer patients between 1998 and 2007 were included in our study. The crude incidence rate was 47.81/100 000 (32 845/68 704 429), increasing by 38.80% from 39.30/100 000 in 1998 to 54.55/100 000 in 2007 with APC at 3.35% in urban Beijing (Z = 9.984, P < 0.001). While it changed to 28.95/100 000 with an APC at 0.27% (Z = 0.846, P = 0.422) when adjusted by world standardized population. For male, the crude incidence rate was 58.28/100 000 (20 342/34 906 580, adjusted rate at 37.03/100 000, APC at 0.38%, Z = 1.008, P = 0.343); while for female, the crude incidence rate was 36.99/100 000 (12 503/33 797 849, adjusted rate at 21.48/100 000, APC at 0.14%, Z = 0.431, P = 0.678). 17 920 lung cancer patients being diagnosed according to histological evidence, accounted for 54.56%. The respective proportion of the patients with histological diagnosis was 43.14% (1095/2538) in 1998 and 65.55% (2641/4029) in 2007, with a 51.95% increase (χ(2) = 859.152, P < 0.001) in decade. In terms of subtypes of lung cancer, the proportion of squamous cell carcinoma decreased annually, from 30.41% (333/1095) in 1998 to 24.16% (638/2641) in 2007; while the proportion of adenocarcinoma increased from 42.83% (469/1095) to 46.80% (1236/2641). As a result, the squamous cell carcinoma to adenocarcinoma ratio declined from 0.71 (333/469) to 0.52 (638/1236) (χ(2) = 50.214, P < 0.001). For women, the ratio declined more significantly and the proportion of the squamous cell carcinoma and adenocarcinoma were 14.77% (925/6262) and 60.83% (3809/6262), respectively in the period between 1998 and 2007. CONCLUSION: No significant change was found in the incidence trend of lung cancer after the incidence rate adjusted by world standard population, but the proportion of the subtypes of lung cancer categorized by histological evaluation changed apparently.

Time trends of cancer incidence in urban beijing, 1998-2007.

OBJECTIVE: To report the time trends of cancer incidence in urban Beijing from 1998 to 2007. METHODS: All data were obtained from Beijing Cancer Registry from 1998-2007 in urban Beijing. Time trends of incidence rate of cancer were assessed by annual percentage change (APC) of age-standardized incidence rate (ASR) of world standard population during this 10-year period. RESULTS: For all 156851 cancer cases combined, the incidence rate rose in both males and females, with an APC of 2.23% and 3.74%, respectively. The incidence rate of upper gastrointestinal cancers declined significantly, with an APC of -2.83% in esophageal cancer and -1.37% in male gastric cancer, while lower gastrointestinal cancer, such as colorectal cancer, increased with an APC of 4.08%. The rates of kidney and bladder cancer increased with an APC of 7.93% and 5.57%, respectively. For women, the APC continued to rise in breast cancer (4.98%), ovary cancer (6.16%), cervix uteri cancer (11.74%), corpus uteri cancer (6.96%) and thyroid gland cancer (13.39%). The rate of prostate cancer among men increased with an APC of 8.58%; the rate of lymphoma and leukemia increased with APC of 5.48% and 5.44%, respectively. CONCLUSION: Because of population aging and urban modernization, the cancer burden in Beijing has become more and more severe and similar to the western developed countries.

[Screening and identification of differential serum proteins related to dermatitis medicamentosa-like of trichloroethylene].

OBJECTIVE: To screen and identify differential serum proteins which might be involved in dermatitis medicamentosa-like of trichloroethylene (DMLT). METHODS: Three groups of sera were collected from population exposed to trichloroethylene (TCE) (group I), patients suffering from DMLT (group II), and the healed cases (group III). After removing albumin and IgG in the three pools of sera, a comparative proteomic analysis was carried out. The images were analyzed using ImageMaster Platinum 2D 5.0 to screen the differentially expressed proteins. The protein spots were then subjected to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and tandem mass spectrometry sequencing of tryptic peptides for further identification. RESULTS: The depletion of albumin and IgG greatly increased the number of protein spots to 300 ± 12.Five differential spots were identified, which were complement component C4b, apolipoprotein A-I, apolipoprotein C-III apolipoprotein C-II and transthyretin. Compared with group I, the expression levels of complement component C4b in group III and apolipoprotein C-II in group II were up-regulated (1.352 88-fold, 1.512 14-fold, respectively); compared with group I, the expression levels of apolipoprotein A-I, apolipoprotein C-III and transthyretin in group II were down-regulated (1.601 17-fold, 1.034 49-fold, 1.313 35-fold, respectively). CONCLUSION: The findings of this study show that most of the identified differential proteins are closely related to immunity and liver dysfunction, which provides some evidence on elucidating the mechanisms and screening of biomarkers of TCE intoxication.

[Screening and identification of differentially expressed proteins between adult female and male worms of Schistosoma japonicum].

OBJECTIVE: To screen and identify differentially expressed proteins between adult female and male worms of Schistosoma japonicum(S.japonicum). METHODS: Two rabbits infected with the cercaria were perfused with saline in carotid, and approximately two hundred adult female and two hundred male worms of S.japonicum were collected. Approximately 300 microg soluble and hydrophobic proteins of adult female and male worms of S.japonicum were extracted and then the proteins were separated by two-dimensional gel electrophoresis respectively. The analysis using ImageMaster Platinum 2D 5.0 resulted in differentially expressed proteins between adult female and male worms, which were subjected to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and tandem mass spectrometry sequencing of tryptic peptides. RESULTS: There were (255 +/- 10) and (224 +/- 12) spots detected for soluble proteins and (200 +/- 11) and (132 +/- 8) spots for hydrophobic proteins from adult female and male worms respectively. Six differential proteins were identified, five up-regulated proteins in female worms were thioredoxin, putative ferritin-1 heavy chain, chain B in solution structure of the human ubiquitin-conjugating-enzyme-like protein Mms2-Ubiquitin Complex, heat shock protein 10, cytoplasmic fatty acid binding protein variant H; while only one up-regulated proteins in male worms was identified as 48 kDa histamine receptor subunit peptide 4. CONCLUSION: Several differentially expressed proteins between female and male worms of S. japonicum were recognized through screening and identifying differential proteins between female and male worms of S.japonicum.

An investigation of hormesis of trichloroethylene in L-02 liver cells by differential proteomic analysis.

Hormesis is the dose-response pattern of the biological responses to toxic chemicals, characterized by low-dose stimulation and high-dose inhibition. Although it is known that some cell types exhibit an adaptive response to low levels of cytotoxic agents, its molecular mechanism is still unclear and it has yet to be established whether this is a universal phenomenon that occurs in all cell types in response to exposure to every chemical. Trichloroethylene (TCE) is an organic solvent widely used and is released into the atmosphere from industrial degreasing operations. Acute (short-term) and chronic (long-term) inhalation exposure to trichloroethylene can affect the human health. In order to elucidate a cell-survival adaptive response of L-02 liver cells exposed to low dose of TCE, CCK-8 assay was used to assess cytotoxicity, and examined the possible mechanisms of hormesis by proteomics technology. We found that exposure of L-02 liver cells to low level of TCE resulted in adaptation to further exposure to higher level, about 1,000 protein-spots were obtained by two-dimensional electrophoresis (2-DE) and five protein spots were identified by matrix-assisted laser desorption/ionization mass spectrometry and tandem mass spectrometry sequencing of tryptic peptides. Our results suggest that a relationship may exist between identified proteins and TCE-induced hormesis, which are very useful for further study of the mechanism and risk assessment of TCE.

[Effect of vasoactive intestinal peptide and epidermal growth factor on the adhesion of eosinophils to bronchial epithelial cells].

OBJECTIVE: To investigate the effect of intrapulmonary regulatory peptides on adhesion of eosinophils (EOS) to bronchial epithelial cells (BECs). METHODS: Two regulatory peptides, namely vasoactive intestinal peptide (VIP) and epidermal growth factor (EGF) were investigated. VIP and EGF were observed on the secretion of ILs and expression of intercellular adhesion molecule-1 (ICAM-1). RESULTS: VIP and EGF could decrease ILs secretion and ICAM-1 expression. CONCLUSION: VIP and EGF inhibited the adhesion of EOS to BEC in the inflammatory process to lighten the airway inflammation.

[Analysis of the incidence and survival of female breast cancer in Beijing during the last 20 years].

OBJECTIVE: To provide scientific evidence for breast cancer prevention and control through epidemiological analysis of the incidence, mortality and survival rate of female breast cancer in Beijing. METHODS: The registration data of females in Beijing urban area from 1982 to 2001 were retrospectively reviewed. The incidence, mortality and survival rate of female breast cancer were analyzed using routine and life table statistical methods. RESULTS: There was a trend of annual increase by an average of 4.6% and 4.9% in the Beijing urban incidence and world population standardized incidence of female breast cancer during the period of 1982 to 2001. The epidemiological features of Beijing urban female breast cancer showed: (1) The incidence curve of different age groups from 25 to 80 years elevated with two peaks at age of >or= 45 and >or= 70 years; (2) There was an elevation in each age group during the last 20 years; (3) The interception rate at age of 35 to 64 reached 95.3/100,000 population, which made the breast cancer become the number one cancer in female. The changes of survival rate showed: the 5-year observed survival rate (OSR) increased from 62.0% in 1982 - 1983 to 68.7% in 1987 - 1988, the relative survival rate (RSR) increased from 66.3% to 74.2%. The OSR and RSR in 1987 - 1988 were 60.3% and 65.1% at 10 years, and 57.7% and 61.3% 15 years, respectively. The mortality rate of breast cancer fluctuated at 8 to 10 per 10(5) population during the last 20 years. CONCLUSION: There is a trend of an annual increase in female breast cancer in Beijing. The 5-year survival is being improved gradually while the mortality rate remains stable. The results demonstrate that the "early prevention, early diagnosis and early treatment" principles for breast cancer is effective in Beijing.