SSR-based association mapping of salt tolerance in cotton (Gossypium hirsutum L.).

The identification of simple sequence repeat (SSR) markers associated with salt tolerance in cotton contributes to molecular assisted selection (MAS), which can improve the efficiency of traditional breeding. In this study, 134 samples of upland cotton cultivars were selected. The seedling emergence rates were tested under 0.3% NaCl stress. A total of 74 SSR markers were used to scan the genomes of these samples. To identify SSR markers associated with salt tolerance, an association analysis was performed between salt tolerance and SSR markers using TASSEL 2.1, based on the analysis of genetic structure using Structure 2.3.4. The results showed that the seedling emergence rates of 134 cultivars were significantly different, and 27 salt-sensitive and 10 salt-tolerant cultivars were identified. A total of 148 loci were found in 74 SSR markers involving 246 allelic variations, which ranged from 2 to 7 with an average of 3.32 per SSR marker. The gene diversity ranged from 0.0295 to 0.4959, with the average being 0.2897. The polymorphic information content ranged from0.0290 to 0.3729, with the average being 0.2381. This natural population was classified into two subgroups by Structure 2.3.4, containing 89 and 45 samples, respectively. Finally, eight SSR sites associated with salt tolerance ware found through an association analysis, with the rate of explanation ranging from 2.91 to 7.82% and an average of 4.32%. These results provide reference data for the use MAS for salt tolerance in cotton.

Assessing the interactions between the associations of common genetic variants on 2q35 and 16q12 with breast cancer risk.

UNLABELLED: Genome-wide association studies (GWAS) have identified 2q35 and 16q12 as breast cancer (BC) susceptibility loci. However, the association between the two polymorphisms and BC remains controversial and inconsistent. We therefore performed a more precise estimation of these relationships by meta-analysing the currently available evidence from the literature. The PubMed, Ovid, Medline and Web of Science databases were searched. Crude odds ratios (ORs) with 95% confidence intervals (CIs) were used to assess the strengths of the associations. Thirty studies, including 106,312 cases and 140,939 controls, were identified. Overall, significantly elevated breast cancer risk was associated with the A allele of 2q35 rs13387042 when all studies were pooled into the meta-analysis (OR 1.11, 95%CI 1.07-1.15). Additionally, the T allele of 16q12 rs3803662 was associated with significantly increased breast cancer risk (OR 1.20, 95%CI 1.16-1.24). When stratifying for ethnicity, significantly increased risks were found among Caucasians, Asians and mixed ethnicities for both rs13387042 and rs3803662. For rs13387042, an association was observed for both oestrogen receptor-positive (ER+) (OR 1.14, 95%CI 1.11-1.17) and ER-negative (ER-) disease (OR 1.05, 95%CI 1.01-1.09) and for progesterone receptor-positive (PR+) (OR 1.16, 95%CI 1.12-1.19) and PR-negative (PR-) disease (OR 1.07, 95%CI 1.03-1.12). Similarly, a stronger association was observed for rs3803662 with ER+ tumors (OR 1.23, 95%CI 1.13-1.32) compared with ER- tumors (OR 1.08, 95%CI 0.97-1.20), and the same condition occurred for the polymorphism with PR+ tumors (OR 1.26, 95%CI 1.02-1.55) versus with PR- tumors (OR 1.15, 95%CI 0.90-1.46). When stratified by BRCA mutation status, a stronger association was observed with BRCA2 carriers (OR 1.23, 95%CI 1.05-1.44) than BRCA1 carriers (OR 1.09, 95%CI 1.04-1.15). In conclusion, this meta-analysis demonstrated that the A allele of 2q35 rs13387042 and the T allele of 16q12 rs3803662 are risk factors associated with increased breast cancer susceptibility. KEYWORDS: rs13387042, rs3803662, oestrogen receptor, progesterone receptor, breast cancer, meta-analysis.

Single nucleotide polymorphisms in the mitochondrial displacement loop and age-at-onset of non-small cell lung cancer.

The associations between single nucleotide polymorphisms (SNPs) in the displacement loop (D-loop) of mitochondrial DNA (mtDNA) and cancer risk and disease outcome have been extensively analyzed. We investigated the association between age-at-onset and SNPs in the mitochondrial D-loop using a population-based series of non-small cell lung cancer (NSCLC) patients. The D-loop region of mtDNA from NSCLC patients was amplified and sequenced. The age-at-onset curve of NSCLC patients was calculated using the Kaplan-Meier method at each SNP site and values were compared using the log-rank test. The SNP sites of nucleotides 200G/A and 16362T/C were identified to determine their association with age-at-onset of NSCLC using the log-rank test. The nucleotide 207G/A was identified for its association with age-at-onset at a borderline significance level (P = 0.060). We found that genetic polymorphisms in the D-loop were predictive markers for age-at-onset in NSCLC patients. Accordingly, the analysis of genetic polymorphisms in the mitochondrial D-loop can be used to identify NSCLC patient subgroups at high risk of early onset.

SET8 expression is associated with overall survival in gastric cancer.

SET8, a member of the SET domain-containing methyl-transferase, has been implicated in various biological processes. In this study, SET8 was immunostained in 100 samples of gastric cancer tissues and semi-quantified using the HSCORE method to determine the predictive value of SET8 expression levels for gastric cancer outcome. The relationship between SET8 expression and the 5-year survival rate of gastric cancer patients was assessed. High expression of SET8 was associated with a shorter survival time in gastric cancer patients, and the level of SET8 expression was found to be an independent predictor of gastric cancer outcome (relative risk = 1.939; 95% confidence interval = 1.025-3.668; P = 0.042). Analysis of SET8 levels may help in the identification of patient subgroups that are at high risk for poor disease outcomes.

Analysis of upland cotton (Gossypium hirsutum) response to Verticillium dahliae inoculation by transcriptome sequencing.

Verticillium wilt is one of the main diseases in cotton (Gossypium hirsutum), severely reduces yield and fiber quality, and is difficult to be con-trolled effectively. At present, the molecular mechanism that confers resistance to this disease is unclear. Transcriptome sequencing is an important method to detect resistance genes, explore metabolic pathways, and study resistance mechanisms. In this study, the transcriptome of a disease-resistant inbred cot-ton line inoculated with Verticillium dahliae was sequenced. A total of 126,402 unigenes were obtained using de novo assembly and data analysis, 99,712 (78.88%) of which were annotated into the Nr, Nt, Swiss-Prot, KEGG, COG, and GO databases. The expression patterns of 16 candidate disease-resis-tance genes showed that some genes were upregulated soon after V. dahliae inoculation and others were upregulated later, which may indicate instanta-neous basal defense and lagged specific defense, respectively. We conducted a preliminary analysis of the transcriptome database, which will contribute to further research regarding the cloning of disease-resistance genes.

Characterization of a New Anastomosis Group (AG-W) of Binucleate Rhizoctonia, Causal Agent for Potato Stem Canker.

Two binucleate Rhizoctonia (BNR) isolates were recovered from potato cankered stems in Heilongjiang Province, northeastern China. Their cultural appearance on potato dextrose agar remained whitish as the cultures aged. White monilioid cells formed in the fluffy aerial hyphae, whereas no sclerotia appeared during the incubation. The two isolates could anastomose with each other, but they failed to anastomose with reference strains of BNR from AG-A to AG-Q, and AG-U. Analyses of restriction fragment length polymorphism (RFLP) of internal transcribed spacer of ribosomal DNA (rDNA-ITS) regions confirmed that these two isolates differed from the reference strains. The phylogenetic tree based on the sequences of rDNA-ITS regions showed that they were located in a distinct clade from other BNR AGs. These collective results suggested that the isolates recovered from potato in this study belonged to a new BNR AG designated as AG-W. Pathogenicity tests under glasshouse conditions revealed that both isolates were able to cause brown, dry, and slightly sunken lesions on potato subterranean stems. To our knowledge, this is the first report of the AG-W causing potato disease in China as well as worldwide.

Optimization of chloroplast microsatellite PCR conditions and primer screening for endangered Rheum officinale, Rheum palmatum, and Rheum tanguticum.

Chloroplast microsatellite primers were developed in order to provide more population genetic information of endangered Rheum officinale, R. palmatum, and R. tanguticum for conservation. The dried roots and rhizomes of these plants are important in traditional Chinese medicine. The results showed that the optimum concentrations of Mg(2+), Taq DNA polymerase, dNTPs, template DNA, and primers in a 25-µL reaction system were 2.0 mM, 1.0 U, 0.10 mM, 20 ng, and 0.8 µM, respectively. Fourteen of 53 primer combinations were chosen for their high clarity and repetition in three species, and their annealing temperatures ranged from 56 to 58°C. These primers and the optimized polymerase chain reaction system may provide a tool for understanding the demography and genetic variation of these endangered plants.

Fetal fibronectin detection for preterm birth prediction.

To study preterm birth prediction based on fetal fibronectin (fFN) in pregnant women, we randomly selected 124 patients. Vaginal posterior fornix secretions were analyzed using fFN quick test strips. Leucorrhea routine samples were collected to detect bacterial vaginosis, mycoplasma, and chlamydia. Delivery data at 7 days, 14 days, 34 weeks, and 37 weeks were documented and the sensitivity, specificity, positive predictive value, and negative predictive value were analyzed. Of the 124 cases, we found 2, 4, 10, and 18 cases of maternity within 7 days, 14 days, 34 weeks, and 37 weeks, respectively. The sensitivity, specificity, positive predictive value, and negative predictive value were as follows: 100, 77.8, 6.9, and 100% for 7 days; 75, 78.3, 10.3, and 98.9% for 14 days; 50.0, 78.9, 17.2, and 94.7% for 34 weeks; 33.3, 78.3, 20.7, and 87.4% for 37 weeks, respectively. Except for 18 preterm births, 23 cases were fFN-positive, 17 cases had lower genital tract infection. Eighty-three cases were fFN-negative, of which 18 cases had the lower genital tract infections. This difference was statistically significant (P < 0.05). Eighteen cases (14.5% of the pregnant women) had preterm birth. Ten cases delivered within 34 weeks. The negative predictive value and recent predictive value of fFN testing were higher; the positive predictive value was limited due to the impact of lower genital tract infection. The fFN-positive patients need timely clinical processing. During the pregnancy, monitoring of fFN changes and early detection of abnormalities help to reduce perinatal morbidity and mortality.

[Summary of tools for assessment of public health emergency response capability].

With the progress of globalization, the public health emergencies represented by major infectious diseases have become a major challenge for the public health management in China. The article briefly describes the emergency response capability assessment tools in China, and introduces two emergency response assessment tools with complete content structure and wide application in the world. Then the advantages and disadvantages of the tools are compared and discussed in order to provide reference for improvement of the assessment tools for public health emergency response capability in China.

Ubiquitins of Bombyx mori nucleopolyhedrovirus and Helicoverpa armigera nucleopolyhedrovirus show distinct subcellular localization in infected cells.

Ubiquitin (UB) is a conserved protein that regulates a number of processes in eukaryotic cells. Nearly all lepidopteran baculoviruses encode UB homologs showing a partial sequence identity with human UB (Hu-UB). In this study, the sequence, predicted 3D-structure and subcellular localization of UB homologs encoded by two different nucleopolyhedroviruses of Bombyx mori (BmNPV) and Helicoverpa armigera (HaNPV) were compared. UBs of BmNPV and HaNPV (Bm-UB, Ha-UB, respectively) shared only 73% of sequence identity of the different aa in relation to Hu-UB being localized in non-conserved parts, namely in two heterogeneous regions of aa 15-32 and aa 53-60. Interestingly, Bm-UB and Ha-UB share the same seven lysines except for an additional Lys54 in Bm-UB. However, in spite of the sequence heterogeneity, Bm-UB and Ha-UB have a similar predicted 3D-structure. A difference in their subcellular localization during virus growth in insect cell lines was found in the late stage of formation of occlusion-derived virus (ODV). In particular Bm-UB was localized mainly and evenly in the nucleus, while Ha-UB on the nuclear membrane. These data suggest that (i) UBs, besides being engaged in various cellular processes, have a role in specific processes of virus growth, and (ii) Bm-UB and Ha-UB may show certain different activities associated with the virus growth.

[Analysis of cases with maxillofacial arteriovenous malformation treated with balloon temporary block and percutaneous puncture sclerotherapy].

To analysis the treatment effectiveness of maxillofacial arteriovenous malformation (AVM) by means of temporary balloon block and percutaneous puncture sclerotherapy. The treatment outcomes of 20 patients with AVM of maxillofacial deformity, aged (26.8±3.0) 18-36 years, from Dec 2012 to Dec 2019 were collected and retrospectively analyzed. All patients had been diagnosed AVM by digital substraction angiography (DSA). All the cases were treated by using balloons to temporary block responsible artery of the AVM in order to change the high-flow malformations into low-flow venous malformation. Then injection of polyethylene foam cinnamyl alcohol was conducted via percutaneous puncturing for hardening the vascular malformation. Comparing the symptoms and imaging changes before treatment with those 1-6 months after treatment according to Achauer curative effect evaluation standard. Totally 60% (12/20) of the cases were type Ⅱa AVM, and their treatments were effective. However, the treatment effects of type Ⅲb and type Ⅳ patients were relatively poor. The clinical effective rate was 85% (17/20), and the clinical curative rate was 50% (10/20). No skin necrosis, organ damage and other serious complications were observed for the 20 patients. The AVM patients with a main supply artery treated by using a balloon artery temporary block to make high flow malformations into low-flow and followed by percutaneous puncture sclerotherapy, could get good curative effect. It seemed that this approach was safe and effective.

Characterization of ORF127 of Helicoverpa armigera nucleopolyhedrovirus.

It has been shown that Ha127 in the genome of Helicoverpa armigera nucleopolyhedrovirus (HaNPV) has homologs in some other baculoviruses and encodes a putative protein of 192 aa. In this study, a sequence analysis showed the transcription initiation site in Ha127 gene at nts 188 upstream of the translation initiation codon ATG and a potential leucine zipper motif at aa 34-55 in the corresponding protein. Ha127 transcripts were detected in HaNPV-infected HzAM1 cells at 18-72 hrs post infection ( p.i.) by RT-PCR, while the corresponding protein was found at 24-72 hrs p.i. by Western blot analysis suggesting that Ha127 is a late gene product. The size of detected Ha127 protein was about 28 K, a larger value than the predicted 22.6 K indicating a post-translational modification. Immunofluorescence assay of HzAM1 cells infected with HaNPV and Ha127-EGFP expression showed that Ha127 protein was localized in the nucleus. In summary, these data suggested that Ha127 was a functional ORF that might play a role in the nucleus during the late or very late gene expression.

[The correlation study between PM2.5 and the life quality in patients with allergic rhinitis].

Objective:This study aims to research the relationship betweeen allergic rhinitis (AR) patients life quality and the PM2.5 concentration. Method:Fifty-two patients with clinically diagnosed AR were enrolled in this study. Patients were asked to fill in the questionnaire about the quality of life of rhinoconjunctivitis (RQLQ) continuously. The concentrations of PM2.5 in their living environment were continuously tested for one month (31 days) and SPSS 19.0 software was used to analyze data through descriptive statistical method, Spearman correlation analysis and nonparametric test. P<0.05 was considered statistically significant. Result:There were significant association between PM2.5 and nasal symptoms (r=0.121, P<0.01), daily activities (r=0.146, P<0.01) and practical problems (r=0.099, P<0.01). However, sleep (r=0.059, P=0.051), non-hay fever symptoms (r=0.042, P=0.169), emotion (r=0.042, P=0.168), eye symptoms (r=0.087, P=0.274) and PM2.5 had no statistical significance. AR patients have faced notable differences in genders. The scores of activities, non hay fever symptoms and emotions also showed the difference. Female AR patients have demonstrated the statistical significance with the concentration of PM2.5 among the activity, non hay fever symptoms, practical problems, nasal symptoms and emotions. While the male AR patients existed a statistical significance in the concentration of PM2.5 only between the activity and nasal symptoms. Conclusion:PM2.5 concentration is negatively associated with the life quality of AR patients. The higher concentration of PM2.5, the lower AR patients quality of life they got.

Performance of tPSA and f/tPSA for prostate cancer in Chinese. A systematic review and meta-analysis.

Performing a systematic review and meta-analysis to evaluate the diagnostic performance of the total prostate-specific antigen (tPSA) tests for diagnosis of prostate cancer among Chinese. Literatures related to diagnosis of prostate cancer by tPSA and the ratio of free to total PSA (f/tPSA) being retrieved in five electronic databases. Sensitivity, specificity, positive likelihood ratio, negative likelihood ratio and diagnostic odds ratio (DOR) were pooled using random effects models. Summary receiver operating characteristic curves (SROC) used to summarize overall test performance. In total 10 studies meeting the inclusion criteria, 2256 patients were included. Among them 423 were patients of prostate cancer compared to 1833 were patients of benign prostate hyperplasia (BPH). The pooled sensitivity and specificity of tPSA>4.0 ng/ml, tPSA>10.0 ng/ml and f/tPSA<0.15 as threshold for the diagnosis of prostate cancer were heterogeneity. The areas under SROC (AUC) were 80, 85 and 87%, respectively (P<0.05). The pooled DOR were 8.44(95%CI: 4.45 approximately 16.00), 9.94(95%CI: 4.15 approximately 23.77) and 13.75(95%CI: 6.35 approximately 29.76), respectively (P<0.05). chi(2) test used for testing the heterogeneity. tPSA and f/tPSA made an important performance for diagnosis of prostate cancer in decade among Chinese. The use of f/tPSA<0.15 as threshold maintain a high prognostic accuracy for prostate cancer.

Aspergillus fumigatus CY018, an endophytic fungus in Cynodon dactylon as a versatile producer of new and bioactive metabolites.

Aspergillus fumigatus CY018 was recognized as an endophytic fungus for the first time in the leaf of Cynodon dactylon. By bioassay-guided fractionation, the EtOAc extract of a solid-matrix steady culture of this fungus afforded two new metabolites, named asperfumoid (1) and asperfumin (2), together with six known bioactive compounds including monomethylsulochrin, fumigaclavine C, fumitremorgin C, physcion, helvolic acid and 5alpha,8alpha-epidioxy-ergosta-6,22-diene-3beta-ol as well as other four known compounds ergosta-4,22-diene-3beta-ol, ergosterol, cyclo(Ala-Leu) and cyclo(Ala-Ile). Through detailed spectroscopic analyses including HRESI-MS, homo- and hetero-nuclear correlation NMR experiments (HMQC, COSY, NOESY and HMBC), the structures of asperfumoid and asperfumin were established to be spiro-(3-hydroxyl-2,6-dimethoxyl-2,5-diene-4-cyclohexone-(1,3')-5'-methoxyl-7'-methyl-(1'H, 2'H, 4'H)-quinoline-2',4'-dione) and 5-hydroxyl-2-(6-hydroxyl-2-methoxyl-4-methylbenzoyl)-3,6-dimethoxyl-benzoic methyl ester, respectively. All of the 12 isolates were subjected to in vitro bioactive assays against three human pathogenic fungi Candida albicans, Tricophyton rubrum and Aspergillus niger. As a result, asperfumoid, fumigaclavine C, fumitremorgin C, physcion and helvolic acid were shown to inhibit C. albicans with MICs of 75.0, 31.5, 62.5, 125.0 and 31.5 microg/mL, respectively.

A serine protease from suspension-cultured soybean cells.

A serine protease was purified from suspension-cultured soybean cells, by a combination of anion exchange, hydrophobic interaction and affinity chromatography. A 90,000 M(r) subunit, which could be photoaffinity labelled with 3H-diisopropylfluorophosphate (DFP), was identified by SDS-polyacrylamide gel electrophoresis. The enzyme had a broad pH optimum from 5.5 to 8.5, and was strongly inhibited by antipain, leupeptin, aminoethylbenzenesulphonyl fluoride (AEBSF) and DFP, but not by soybean trypsin inhibitor. It cleaved several peptide 4-methylcoumaryl-7-amide derivatives after arginine or lysine residues. Mass spectroscopic analysis of oligopeptide digestion products indicated that the preferred cleavage positions were between paired arginine residues, or C-terminal to single arginine residues, depending on the oligopeptide substrate. Partial amino acid sequences from the purified protein showed sequence identity to bacterial protease II and prolyl peptidase, although the enzyme lacked prolyl endopeptidase activity. We discuss the possible involvement of the protease in plant defense responses.

Effects of calcium and calmodulin inhibitors on the abnormal proliferation of lung fibroblast.

The effects of alveolar macrophage (Am phi) conditioned media from interstitial lung disease (ILD) patients on fibroblast (FB), and the role of calcium (Ca2+) blockers and calmodulin (CaM) inhibitors on the proliferation of lung FB were studied. We found that the AM phi conditioned media could stimulate FB cell proliferation and this effect could be abolished by Ca2+ blockers and CaM inhibitors. The results indicated that AM phi was in activated state in ILD and released some kinds of cytokines to stimulate the proliferation of FB, and Ca,2+ CaM were partially responsible for these actions.

[Effects of calcium blockers and calmodulin inhibitors on the proliferation of fibroblast in alveolar macrophage condition medium].

Alveolar macrophages (AM phi) play a key role in the process of fibrosis in many lung diseases. We investigated the effects of AM phi on the proliferation of fibroblast(FB) in various lung diseases and the influence of calcium and CaM inhibitors on those effects. AM phi conditioned media were prepared from 17 patients with interstitial lung diseases (ILD) and 13 patients with bronchogenic carcinoma (BC). In ILD group, conditioned media have stimulating effect on the proliferation of FB (P < 0.05), this effect was inhibited markedly by calcium channel blocker verapamil (2.5 x 10(-2)mg/ml, 2.5 x 10(-4)mg/ml), diltiazem (5 x 10(-2)mg/ml) and partially by CaM inhibitor chlorpromazine (2.5 x 10(-4)mg/ml). Whereas in BC group, only conditioned media from AM phi in the presence of stimulant have marked stimulating effect on the proliferation of FB, and this effect was markedly inhibited both by Ca++ channel blocker verapamil (2.5 x 10(-4)mg/ml), diltiazem (5 x 10(-4)mg/ml) and CaM inhibitor chlorpromazine (1.25 x 10(-2)mg/ml, 2.5 x 10(-4)mg/ml). We conclude that AM phi are activated in patients with ILD, but not in those with BC. The stimulating effect of cytokines secreted by activated AM phi can be inhibited by Ca blocker and CaM inhibitor. Using Ca++ and CaM blockers may be a new and promising approach in the treatment of ILD.

Search for the invisible decay of J/psi in psi(2S) --> pi(+)pi(-) J/psi.

Using psi(2S) --> pi(+)pi(-) J/psi events in a sample of 14.0 x 10(6) psi(2S) decays collected with the BES-II detector, a search for the decay of the J/psi to invisible final states is performed. No signal is found, and an upper limit at the 90% confidence level is determined to be 1.2 x 10(-2) for the ratio B(J/psi --> invisible)/B(J/psi-->mu(+)mu(-)). This is the first search for J/psi decays to invisible final states.