Molecular Mechanism of Different Rooting Capacity between Two Clones of Taxodium hybrid 'Zhongshanshan'.

The conifer Taxodium hybrid 'Zhongshanshan' (T. hybrid 'Zhongshanshan') is characterized by rapid growth, strong stress resistance, and high ornamental value and has significant potential for use in afforestation, landscaping, and wood production. The main method of propagating T. hybrid 'Zhongshanshan' is tender branch cutting, but the cutting rooting abilities of different T. hybrid 'Zhongshanshan' clones differ significantly. To explore the causes of rooting ability differences at a molecular level, we analyzed the transcriptome data of cutting base and root tissues of T. hybrid 'Zhongshanshan 149' with a rooting rate of less than 5% and T. hybrid 'Zhongshanshan 118' with rooting rate greater than 60%, at the developmental time points in this study. The results indicated that differentially expressed genes between the two clones were mainly associated with copper ion binding, peroxidase, and oxidoreductase activity, response to oxidative stress, phenylpropanoid and flavonoid biosynthesis, and plant hormone signal transduction, among others. The expression pattern of ThAP2 was different throughout the development of the adventitive roots of the two clone cuttings. Therefore, this gene was selected for further study. It was shown that ThAP2 was a nuclear-localized transcription factor and demonstrated a positive feedback effect on rooting in transgenic Nicotiana benthamiana cuttings. Thus, the results of this study explain the molecular mechanism of cutting rooting and provide candidate gene resources for developing genetic breeding strategies for optimizing superior clones of T. hybrid 'Zhongshanshan'.

Development of an UV-Resistant Multilayer Film with Enhanced Compatibility between Carboxymethyl Cellulose and Polylactic Acid via Incorporation of Tannin and Ferric Chloride.

Carboxymethyl cellulose (CMC) and polylactic acid (PLA) are recognized for their environmental friendliness. By merging them into a composite film, packaging solutions can be designed with good performance. Nonetheless, the inherent interface disparity between CMC and PLA poses a challenge, and there may be layer separation issues. This study introduces a straightforward approach to mitigate this challenge by incorporating tannin acid and ferric chloride in the fabrication of the CMC-PLA. The interlayer compatibility was improved by the in situ formation of a cohesive interface. The resulting CMC/TA-PLA/Fe multilayer film, devoid of any layer separation, exhibits exceptional mechanical strength, with a tensile strength exceeding 70 MPa, a high contact angle of 105°, and superior thermal stability. Furthermore, the CMC/TA-PLA/Fe film demonstrates remarkable efficacy in blocking ultraviolet light, effectively minimizing the discoloration of various wood surfaces exposed to UV aging.

Comparative chloroplast genomics of the genus Taxodium.

BACKGROUND: Chloroplast (cp) genome information would facilitate the development and utilization of Taxodium resources. However, cp genome characteristics of Taxodium were poorly understood. RESULTS: We determined the complete cp genome sequences of T. distichum, T. mucronatum, and T. ascendens. The cp genomes are 131,947 bp to 132,613 bp in length, encode 120 genes with the same order, and lack typical inverted repeat (IR) regions. The longest small IR, a 282 bp trnQ-containing IR, were involved in the formation of isomers. Comparative analysis of the 3 cp genomes showed that 91.57% of the indels resulted in the periodic variation of tandem repeat (TR) motifs and 72.46% single nucleotide polymorphisms (SNPs) located closely to TRs, suggesting a relationship between TRs and mutational dynamics. Eleven hypervariable regions were identified as candidates for DNA barcode development. Hypothetical cp open reading frame 1(Ycf1) was the only one gene that has an indel in coding DNA sequence, and the indel is composed of a long TR. When extended to cupressophytes, ycf1 genes have undergone a universal insertion of TRs accompanied by extreme length expansion. Meanwhile, ycf1 also located in rearrangement endpoints of cupressophyte cp genomes. All these characteristics highlight the important role of repeats in the evolution of cp genomes. CONCLUSIONS: This study added new evidence for the role of repeats in the dynamics mechanism of cp genome mutation and rearrangement. Moreover, the information of TRs and hypervariable regions would provide reliable molecular resources for future research focusing on the infrageneric taxa identification, phylogenetic resolution, population structure and biodiversity for the genus Taxodium and Cupressophytes.

An Integrated Transcriptome and Proteome Analysis Reveals Putative Regulators of Adventitious Root Formation in Taxodium 'Zhongshanshan'.

Adventitious root (AR) formation from cuttings is the primary manner for the commercial vegetative propagation of trees. Cuttings is also the main method for the vegetative reproduction of Taxodium 'Zhongshanshan', while knowledge of the molecular mechanisms regulating the processes is limited. Here, we used mRNA sequencing and an isobaric tag for relative and absolute quantitation-based quantitative proteomic (iTRAQ) analysis to measure changes in gene and protein expression levels during AR formation in Taxodium 'Zhongshanshan'. Three comparison groups were established to represent the three developmental stages in the AR formation process. At the transcript level, 4743 genes showed an expression difference in the comparison groups as detected by RNA sequencing. At the protein level, 4005 proteins differed in their relative abundance levels, as indicated by the quantitative proteomic analysis. A comparison of the transcriptome and proteome data revealed regulatory aspects of metabolism during AR formation and development. In summary, hormonal signal transduction is different at different developmental stages during AR formation. Other factors related to carbohydrate and energy metabolism and protein degradation and some transcription factor activity levels, were also correlated with AR formation. Studying the identified genes and proteins will provide further insights into the molecular mechanisms controlling AR formation.

High-density genetic map construction and quantitative trait loci identification for growth traits in (Taxodium distichum var. distichum × T. mucronatum) × T. mucronatum.

BACKGROUND: 'Zhongshanshan' is the general designation for the superior interspecific hybrid clones of Taxodium species, which is widely grown for economic and ecological purposes in southern China. Growth is the priority objective in 'Zhongshanshan' tree improvement. A high-density linkage map is vital to efficiently identify key quantitative trait loci (QTLs) that affect growth. RESULTS: In total, 403.16 Gb of data, containing 2016,336 paired-end reads, was obtained after preprocessing. The average sequencing depth was 28.49 in T. distichum var. distichum, 25.18 in T. mucronatum, and 11.12 in each progeny. In total, 524,662 high-quality SLAFs were detected, of which 249,619 were polymorphic, and 6166 of the polymorphic markers met the requirements for use in constructing a genetic map. The final map harbored 6156 SLAF markers on 11 linkage groups, and was 1137.86 cM in length, with an average distance of 0.18 cM between adjacent markers. Separate QTL analyses of traits in different years by CIM detected 7 QTLs. While combining multiple-year data, 13 QTLs were detected by ICIM. 5 QTLs were repeatedly detected by the two methods, and among them, 3 significant QTLs (q6-2, q4-2 and q2-1) were detected in at least two traits. Bioinformatic analysis discoveried a gene annotated as a leucine-rich repeat receptor-like kinase gene within q4-2. CONCLUSIONS: This map is the most saturated one constructed in a Taxodiaceae species to date, and would provide useful information for future comparative mapping, genome assembly, and marker-assisted selection.

Estimating lead-time bias in lung cancer diagnosis of patients with previous cancers.

Surprisingly, survival from a diagnosis of lung cancer has been found to be longer for those who experienced a previous cancer than for those with no previous cancer. A possible explanation is lead-time bias, which, by advancing the time of diagnosis, apparently extends survival among those with a previous cancer even when they enjoy no real clinical advantage. We propose a discrete parametric model to jointly describe survival in a no-previous-cancer group (where, by definition, lead-time bias cannot exist) and in a previous-cancer group (where lead-time bias is possible). We model the lead time with a negative binomial distribution and the post-lead-time survival with a linear spline on the logit hazard scale, which allows for survival to differ between groups even in the absence of bias; we denote our model Logit-Spline/Negative Binomial. We fit Logit-Spline/Negative Binomial to a propensity-score matched subset of the Surveillance, Epidemiology, and End Results-Medicare linked data set, conducting sensitivity analyses to assess the effects of key assumptions. With lung cancer-specific death as the end point, the estimated mean lead time is roughly 11 months for stage I&II patients; with overall survival, it is roughly 3.4 months in stage I&II. For patients with higher-stage lung cancers, the mean lead time is 1 month or less for both outcomes. Accounting for lead-time bias reduces the survival advantage of the previous-cancer group when one exists, but it does not nullify it in all cases.

Revisiting a longstanding clinical trial exclusion criterion: impact of prior cancer in early-stage lung cancer.

BACKGROUND: Early-stage lung cancer represents a key focus of numerous multicenter clinical trials, but common exclusion criteria such as a prior cancer diagnosis may limit enrollment. We examined the prevalence and prognostic impact of a prior cancer diagnosis among patients with early-stage lung cancer. METHODS: We identified patients>65 years of age with early-stage lung cancer diagnosed 1996-2009 in the Surveillance, Epidemiology, and End Results-Medicare linked database. Prior cancers were characterized by type, stage, and timing with respect to the lung cancer diagnosis. All-cause and lung cancer specific-survival rates were compared between patients with and without prior cancer using Cox regression analyses and propensity scores. RESULTS: Among 42,910 patients with early-stage lung cancer, one-fifth (21%) had a prior cancer. The most common prior cancers were prostate (21%), breast (18%), gastrointestinal (17%), and other genitourinary (15%). Most prior cancers were localized, and 61% were diagnosed within 5 years of the lung cancer diagnosis. There was no difference in all-cause survival between patients with and without prior cancer (hazard ratio [HR] 1.01; P=0.52). Lung cancer specific survival was improved among patients with prior cancer (HR 0.79; P<0.001). CONCLUSIONS: A prior cancer history may exclude a substantial proportion of patients with early-stage lung cancer from enrollment in clinical trials. Without adverse effect on clinical outcomes, inclusion of patients age >65 years with prior cancer in clinical trials should be considered to improve study accrual, completion rates, and generalizability.

Who Is Food Insecure? Implications for Targeted Recruitment and Outreach, National Health and Nutrition Examination Survey, 2005-2010.

INTRODUCTION: Food insecurity is negatively associated with health; however, health needs may differ among people participating in food assistance programs. Our objectives were to characterize differences in health among people receiving different types of food assistance and summarize strategies for targeted recruitment and outreach of various food insecure populations. METHODS: We examined health status, behaviors, and health care access associated with food insecurity and receipt of food assistance among US adults aged 20 years or older using data from participants (N = 16,934) of the National Health and Nutrition Examination Survey from 2005 through 2010. RESULTS: Food insecurity affected 19.3% of US adults (95% confidence interval, 17.9%-20.7%). People who were food insecure reported poorer health and less health care access than those who were food secure (P < .001 for all). Among those who were food insecure, 58.0% received no assistance, 20.3% received only Supplemental Nutrition Assistance Program (SNAP) benefits, 9.7% received only food bank assistance, and 12.0% received both SNAP and food bank assistance. We observed an inverse relationship between receipt of food assistance and health and health behaviors among the food insecure. Receipt of both (SNAP and food bank assistance) was associated with the poorest health; receiving no assistance was associated with the best health. For example, functional limitations were twice as prevalent among people receiving both types of food assistance than among those receiving none. CONCLUSION: Receipt of food assistance is an overlooked factor associated with health and has the potential to shape future chronic disease prevention efforts among the food insecure.

Residential racial segregation and mortality among black, white, and Hispanic urban breast cancer patients in Texas, 1995 to 2009.

BACKGROUND: The authors investigated whether residential segregation (the degree to which racial/ethnic groups live separately from one another in a geographic area) 1) was associated with mortality among urban women with breast cancer, 2) explained racial/ethnic disparities in mortality, and 3) whether its association with mortality varied by race/ethnicity. METHODS: Using Texas Cancer Registry data, all-cause mortality and breast-cancer mortality were examined among 109,749 urban black, Hispanic, and white women aged ≥50 years who were diagnosed with breast cancer from 1995 to 2009. Racial (black) segregation and ethnic (Hispanic) segregation of patient's neighborhoods were measured and were compared with the larger metropolitan statistical area using the location quotient measure. Shared frailty Cox proportional hazard models were used to nest patients within residential neighborhoods (census tract) and were controlled for race/ethnicity, age, diagnosis year, tumor stage, grade, histology, neighborhood poverty, and county-level mammography availability. RESULTS: Greater black segregation and Hispanic segregation were adversely associated with cause-specific mortality and all-cause mortality. For example, in adjusted models, Hispanic segregation was associated with cause-specific mortality (adjusted hazard ratio, 1.24; 95% confidence interval, 1.05-1.46). Compared with whites, blacks had higher mortality for both outcomes, whereas Hispanics demonstrated equivalent (cause-specific) or lower (all-cause) mortality. Segregation did not explain racial/ethnic disparities in mortality. Within each race/ethnicity strata, segregation was either adversely associated with mortality or was not significant. CONCLUSIONS: Among urban women with breast cancer in Texas, segregation has an independent, adverse association with mortality, and the effect of segregation varies by patient race/ethnicity. The novel application of a small-area measure of relative racial segregation should be examined in other cancer types with documented racial/ethnic disparities across varied geographic areas.

National trends in incidence rates of hospitalization for stroke in children with sickle cell disease.

BACKGROUND: The success of primary stroke prevention for children with sickle cell disease (SCD) throughout the United States is unknown. Therefore, we aimed to generate national incidence rates of hospitalization for stroke in children with sickle cell disease (SCD) before and after publication of the Stroke Prevention Trial in Sickle Cell Anemia (STOP trial) in 1998. PROCEDURE: We performed a retrospective trend analysis of the 1993-2009 Nationwide Inpatient Sample and Kids' Inpatient Databases. Hospitalizations for SCD patients 0-18 years old with stroke were identified by ICD-9CM code. The primary outcome, the trend in annual incidence rate of hospitalization for stroke in children with SCD, was analyzed by linear regression. Incidence rates of hospitalization for stroke before and after 1998 were compared by the Wilcoxon rank-sum test. RESULTS: From 1993 to 2009, 2,024 hospitalizations were identified for stroke. Using the mean annual incidence rate of hospitalization for stroke from 1993 to 1998 as the baseline, the rate decreased from 1993 to 2009 (point estimate = -0.022/100 patient years [95% CI, -0.039, -0.005], P = 0.027). The mean annual incidence rate of hospitalization stroke decreased by 45% from 0.51 per 100 patient years in 1993-1998 to 0.28 per 100 patient years in 1999-2009 (P = 0.008). Total hospital days and charges attributed to stroke also decreased by 45% and 24%, respectively. CONCLUSIONS: After publication of the STOP trial and hydroxyurea licensure in 1998, the incidence of hospitalization for stroke in children with SCD decreased across the United States, suggesting that primary stroke prevention has been effective nationwide, but opportunity for improvement remains.

Language-specific noun bias: evidence from bilingual children.

Most evidence concerning cross-linguistic variation in noun bias, the preponderance of nouns in early expressive lexicons (Gentner, 1982), has come from comparisons of monolingual children acquiring different languages. Such designs are susceptible to a number of potential confounders, including group differences in developmental level and sociodemographic characteristics. The aim of this study was to quantify noun bias in bilingual Mandarin-English toddlers whose expressive lexicons in each language contained 50-300 words. Parents of fifty children (1;10-2;6) reported separately on their English and Mandarin expressive lexicons. The mean percentage of Mandarin nouns (38%) was significantly lower than the percentage of English nouns (54%) and was robust to analyses of twelve potential covariates. Analyses of the most frequently reported words suggested that lexical reduplication could be considered as a potential influence on vocabulary composition in future studies. Results suggest that characteristics of the input significantly shape early lexicons.

ADH Gene Cloning and Identification of Flooding-Responsive Genes in Taxodium distichum (L.) Rich.

As a flooding-tolerant tree species, Taxodium distichum has been utilized in afforestation projects and proven to have important value in flooding areas. Alcohol dehydrogenase (ADH), which participates in ethanol fermentation, is essential for tolerance to the anaerobic conditions caused by flooding. In a comprehensive analysis of the ADH gene family in T. distichum, TdADHs were cloned on the basis of whole-genome sequencing, and then bioinformatic analysis, subcellular localization, and gene expression level analysis under flooding were conducted. The results show that the putative protein sequences of 15 cloned genes contained seven TdADHs and eight TdADH-like genes (one Class III ADH included) that were divided into five clades. All the sequences had an ADH_N domain, and except for TdADH-likeE2, all the other genes had an ADH_zinc_N domain. Moreover, the TdADHs in clades A, B, C, and D had a similar motif composition. Additionally, the number of TdADH amino acids ranged from 277 to 403, with an average of 370.13. Subcellular localization showed that, except for TdADH-likeD3, which was not expressed in the nucleus, the other genes were predominantly expressed in both the nucleus and cytosol. TdADH-likeC2 was significantly upregulated in all three organs (roots, stems, and leaves), and TdADHA3 was also highly upregulated under 24 h flooding treatment; the two genes might play key roles in ethanol fermentation and flooding tolerance. These findings offer a comprehensive understanding of TdADHs and could provide a foundation for the molecular breeding of T. distichum and current research on the molecular mechanisms driving flooding tolerance.

Novel molecular markers for Taxodium breeding from the chloroplast genomes of four artificial Taxodium hybrids.

Taxodium "Zhongshanshan" are a group of intraspecific Taxodium hybrids with superparental dominance and high ecological and economic value in southern China. Identifying the parentage of hybrids, especially the male parent, is critically important for genetic studies. However, the large nuclear genomes of members of the genus Taxodium pose a major challenge for the development of molecular markers. Here, we developed novel molecular markers by conducting a comparative analysis of the chloroplast genomes of four artificial Taxodium hybrids and their parents. The lengths of the whole chloroplast genome ranged from 131,942 to 132,128 bp, and the total guanine (GC) content of the chloroplast genomes ranged from 34.6% to 35.81%. A total of 120 unique genes were identified, including 83 protein-coding genes, 33 transfer RNAs, and four ribosomal RNAs. There were 69-71 simple sequence repeats were detected in the four hybrids. Phylogenetic analysis revealed that these hybrids clustered with their paternal parents. Similar findings were obtained by analysis of the GC content of protein-coding genes. Molecular markers were developed using the highly variable regions of the chloroplast genomes, and polymerase chain reaction (PCR) assays revealed that these markers were effective for identifying the male parents of these hybrids. Our findings indicate for the first time that the chloroplast genomes of Taxodium are paternally inherited. Generally, these molecular markers could facilitate breeding and genetic studies of Taxodium.

Modestly increased use of colonoscopy when copayments are waived.

BACKGROUND & AIMS: Colorectal cancer (CRC) screening with colonoscopy often requires expensive copayments from patients. The 2010 Patient Protection and Affordable Care Act mandated elimination of copayments for CRC screening, including colonoscopy, but little is known about the effects of copayment elimination on use. The University of Texas employee, retiree, and dependent health plan instituted and promoted a waiver of copayments for screening colonoscopies in fiscal year (FY) 2009; we examined the effects of removing cost sharing on colonoscopy use. METHODS: We conducted a retrospective cohort study of 59,855 beneficiaries of the University of Texas employee, retiree, and dependent health plan, associated with 16 University of Texas health and nonhealth campuses, ages 50-64 years at any point in FYs 2002-2009 (267,191 person-years of follow-up evaluation). The primary outcome was colonoscopy incidence among individuals with no prior colonoscopy. We compared the age- and sex-standardized incidence ratios for colonoscopy in FY 2009 (after the copayment waiver) with the expected incidence for FY 2009, based on secular trends from years before the waiver. RESULTS: The annual incidence of colonoscopy increased to 9.5% after the copayment was waived, compared with an expected incidence of 8.0% (standardized incidence ratio, 1.18; 95% confidence interval, 1.14-1.23; P < .001). After adjusting for age, sex, and beneficiary status, the copayment waiver remained significantly associated with greater use of colonoscopy, with an adjusted hazard ratio of 1.19 (95% confidence interval, 1.12-1.26). CONCLUSIONS: Waiving copayments for colonoscopy screening results in a statistically significant, but modest (1.5%), increase in use. Additional strategies beyond removing financial disincentives are needed to increase use of CRC screening.

Hospitalization for invasive pneumococcal disease in a national sample of children with sickle cell disease before and after PCV7 licensure.

OBJECTIVE: To estimate national hospitalization rates for invasive pneumococcal disease (IPD) in children with sickle cell disease (SCD) before and after the 2000 licensure of the heptavalent pneumococcal conjugate vaccine (PCV7). PROCEDURE: We performed a retrospective trend analysis of the 1994-2007 Nationwide Inpatient Sample databases. Hospitalizations involving children with SCD and IPD were identified by ICD-9CM code. The primary outcomes, the annual hospitalization rate for IPD in children with SCD and the proportion of hospitalizations for IPD per 100 total SCD hospitalizations, were analyzed using multivariable linear regression and contingency analysis, respectively. RESULTS: A total of 1,242 hospitalizations for IPD in SCD patients were identified from 1994-2007, with a mortality rate of 2.4%. The national mean annual rate of IPD hospitalization decreased by 65%, from 131.8 cases/year from 1994 to 2000 to 45.5 cases/year from 2001 to 2007 (P = 0.001). The national proportion of hospitalizations for IPD per 100 total SCD hospitalizations decreased from 0.4 to 0.15 (P < 0.0001) over the same interval. Following PCV7 licensure, the mean annual cumulative hospital days and cumulative hospital charges decreased nationally by 53% and 36%, respectively. CONCLUSION: In a national sample, PCV7 licensure is temporally associated with a nearly threefold reduction in IPD hospitalizations in children with SCD.

Andrographolide Inhibits Proliferation and Promotes Apoptosis in Bladder Cancer Cells by Interfering with NF- κ B and PI3K/AKT Signaling In Vitro and In Vivo.

OBJECTIVE: To explore the influences of andrographolide (Andro) on bladder cancer cell lines and a tumor xenograft mouse model bearing 5637 cells. METHODS: For in vitro experiments, T24 cells were stimulated with Andro (0-40 µmol/L) and 5637 cells were stimulated with Andro (0 to 80 µmol/L). Cell growth, migration, and infiltration were assessed using cell counting kit-8, colony formation, wound healing, and transwell assays. Apoptosis rate was examined using flow cytometry. In in vivo study, the antitumor effect of Andro (10 mg/kg) was evaluated by 5637 tumor-bearing mice, and levels of nuclear factor κ B (NF- κ B) and phosphoinositide 3-kinase/AKT related-proteins were determined by immunoblotting. RESULTS: Andro suppressed growth, migration, and infiltraion of bladder cancer cells (P⩽0.05 or P⩽0.01). Additionally, Andro induced intrinsic mitochondria-dependent apoptosis in bladder cancer cell lines. Furthermore, Andro inhibited bladder cancer growth in mice (P⩽0.01). The expression of p65, p-AKT were suppressed by Andro treatment in vitro and in vivo (P⩽0.05 or P⩽0.01). CONCLUSIONS: Andrographolide inhibits proliferation and promotes apoptosis in bladder cancer cells by interfering with NF- κ B and PI3K/AKT signaling in vitro and in vivo.

Medicinal plants and natural compounds against acyclovir-resistant HSV infections.

Herpes simplex virus (HSV), an alphaherpesvirus, is highly prevalent in the human population and is known to cause oral and genital herpes and various complications. Represented by acyclovir (ACV), nucleoside analogs have been the main clinical treatment against HSV infection thus far. However, due to prolonged and excessive use, HSV has developed ACV-resistant strains. Therefore, effective treatment against ACV-resistant HSV strains is urgently needed. In this review, we summarized the plant extracts and natural compounds that inhibited ACV-resistant HSV infection and their mechanism of action.

The complete chloroplast genome sequence of Taxodium ascendens × T. mucronatum hybrid (Cupressaceae).

Taxodium 'Zhongshanshan 401' is an important economic plant with ornamental and ecological values, and has been widely planted in southeastern China. In this study, the complete chloroplast (cp) genome of T. 'Zhongshanshan 401' was sequenced and illustrated to add the more genetic information. The entire cp genome of T. 'Zhongshanshan 401' was 132,037 bp in length with 35.3% overall GC content. The cp genome contained 120 genes, including 83 protein-coding genes, 33 tRNA genes, and four rRNA genes. Fifteen genes contain two exons and two contains three exons. Phylogenetic analysis based on whole cp genome sequences showed that T. 'Zhongshanshan 401' was more closely related to T. mucronatum.