The development and validation of a nomogram-based risk prediction model for mortality among older adults.

Objective: This research aims to construct and authenticate a comprehensive predictive model for all-cause mortality, based on a multifaceted array of risk factors. Methods: The derivation cohort for this study was the Chinese Longitudinal Healthy Longevity Survey (CLHLS), while the Healthy Ageing and Biomarkers Cohort Study (HABCS) and the China Health and Retirement Longitudinal Study (CHARLS) were used as validation cohorts. Risk factors were filtered using lasso regression, and predictive factors were determined using net reclassification improvement. Cox proportional hazards models were employed to establish the mortality risk prediction equations, and the model's fit was evaluated using a discrimination concordance index (C-index). To evaluate the internal consistency of discrimination and calibration, a 10x10 cross-validation technique was employed. Calibration plots were generated to compare predicted probabilities with observed probabilities. The prediction ability of the equations was demonstrated using nomogram. Results: The CLHLS (mean age 88.08, n = 37074) recorded 28158 deaths (179683 person-years) throughout the course of an 8-20 year follow-up period. Additionally, there were 1384 deaths in the HABCS (mean age 86.74, n = 2552), and 1221 deaths in the CHARLS (mean age 72.48, n = 4794). The final all-cause mortality model incorporated demographic characteristics like age, sex, and current marital status, as well as functional status indicators including cognitive function and activities of daily living. Additionally, lifestyle factors like past smoking condition and leisure activities including housework, television viewing or radio listening, and gardening work were included. The C-index for the derivation cohort was 0.728 (95% CI: 0.724-0.732), while the external validation results for the CHARS and HABCS cohorts were 0.761 (95% CI: 0.749-0.773) and 0.713 (95% CI: 0.697-0.729), respectively. Conclusion: This study introduces a reliable, validated, and acceptable mortality risk predictor for older adults in China. These predictive factors have potential applications in public health policy and clinical practice.

HSV-1 selectively packs the transcription factor Oct-1 into EVs to facilitate its infection.

HSV-1 hijacks the cellular vesicular secretion system and promotes the secretion of extracellular vesicles (EVs) from infected cells. This is believed to facilitate the maturation, secretion, intracellular transportation and immune evasion of the virus. Intriguingly, previous studies have shown that noninfectious EVs from HSV-1-infected cells exert antiviral effects on HSV-1 and have identified host restrictive factors, such as STING, CD63, and Sp100 packed in these lipid bilayer-enclosed vesicles. Octamer-binding transcription factor-1 (Oct-1) is shown here to be a pro-viral cargo in non-virion-containing EVs during HSV-1 infection and serves to facilitate virus dissemination. Specifically, during HSV-1 infection, the nuclear localized transcription factor Oct-1 displayed punctate cytosolic staining that frequently colocalized with VP16 and was increasingly secreted into the extracellular space. HSV-1 grown in cells bereft of Oct-1 (Oct-1 KO) was significantly less efficient at transcribing viral genes during the next round of infection. In fact, HSV-1 promoted increased exportation of Oct-1 in non-virion-containing EVs, but not the other VP16-induced complex (VIC) component HCF-1, and EV-associated Oct-1 was promptly imported into the nucleus of recipient cells to facilitate the next round of HSV-1 infection. Interestingly, we also found that EVs from HSV-1-infected cells primed cells for infection by another RNA virus, vesicular stomatitis virus. In summary, this investigation reports one of the first pro-viral host proteins packed into EVs during HSV-1 infection and underlines the heterogenetic nature and complexity of these noninfectious double-lipid particles.

Integrative analysis of the metabolome and transcriptome reveals the molecular regulatory mechanism of isoflavonoid biosynthesis in Ormosia henryi Prain.

Flavonoids are important components of many phytopharmaceuticals, however, most studies on flavonoids and isoflavonoids have been conducted on herbaceous plants of the family Leguminosae, such as soybean, and less attention has been paid to woody plants. To fill this gap, we characterized the metabolome and transcriptome of five plant organs of Ormosia henryi Prain (OHP), a woody Leguminosae plant with great pharmaceutical value. Our results indicate that OHP possesses a relatively high content of isoflavonoids as well as significant diversity, with greater diversity of isoflavonoids in the roots. Combined with transcriptome data, the pattern of isoflavonoid accumulation was found to be highly correlated with differential expression genes. Furthermore, the use of trait-WGCNA network analysis identified OhpCHSs as a probable hub enzyme that directs the downstream isoflavonoid synthesis pathway. Transcription factors, such as MYB26, MYB108, WRKY53, RAV1 and ZFP3, were found to be involved in the regulation of isoflavonoid biosynthesis in OHP. Our findings will be beneficial for the biosynthesis and utilization of woody isoflavonoids.

In Vitro Propagation and Genetic Uniformity Assessment of Manglietiastrum sinicum: A Critically Endangered Magnoliaceae Species.

Manglietiastrum sinicum Y.W. Law is a critically endangered species with great ornamental and commercial value, which urgently requires protection. We tested different combinations of basal media and plant growth regulators to determine (i) the optimal conditions for bud induction and proliferation of explants and (ii) optimal rooting conditions. RAPD- and ISSR-PCR were used to assess the genetic fidelity of regenerated plantlets. Murashige and Skoog medium (MS) supplemented with 0.5 mg/L 6-benzyladenine (BA) and 0.05 mg/L indole-3-butyric acid (IBA) is the optimal medium for bud induction (100% induction). MSM medium (a special basal medium for M. sinicum) was more suitable for the efficient proliferation and rooting of M. sinicum. Maximum bud proliferation rate (446.20%) was obtained on MSM, with 0.4 mg/L BA, 0.5 mg/L kinetin, and 0.06 mg/L IBA, while maximum root induction rate (88.89%) was obtained on MSM supplemented with 0.4 mg/L 1-naphthylacetic acid and 1.0 mg/L IBA with a 7-day initial darkness treatment. The rooted plantlets were transferred to a substrate containing peat soil, perlite, coconut chaff, and bark (volume ratio 2:1:1:1), with a resulting survival rate of 92.2%. RAPD and ISSR markers confirmed the genetic uniformity and stability of regenerated plants.

Case series: O-arm navigation assisted by the Wiltse approach improves the accuracy of pedicle screw placement in ankylosing spondylitis combined with thoracolumbar fractures.

Here we assessed the accuracy of O-arm navigation assisted by Wiltse approach to improve based pedicle screw insertion in ankylosing spondylitis combined with thoracolumbar fractures. We then compared it with the freehand pedicle screw insertion technique. The study sample included 32 patients with ankylosing spondylitis combined with thoracolumbar fractures. Pedicle screw reduction and internal fixation was performed under an O-arm navigation system assisted by a Wiltse approach-combined osteotomy ("navigation group," n = 17) and posterior pedicle screw reduction and internal fixation was performed using freehand technique combined osteotomy ("freehand group," n = 15). We then compared the operation time and bleeding volume between the 2 groups. The visual analog scale (VAS) and Oswestry disability index (ODI) were then used to evaluate the clinical efficacy and the kyphosis Cobb angle was used to evaluate the radiological efficacy before operation, 3 days after operation and after the last follow-up. All complications were noted when detected. Finally, classification of screw positions as proposed by Neo et al was used to evaluate the relationship of the position between the screw, the bone cortex, and the incidence of screw penetration. All patients were followed up for 18 to 36 months (i.e., 24.2 ±â€…3.5 months). The operation time and intraoperative bleeding volume of the navigation group were significantly shorter (lower) than those of the freehand group (P < .05). In addition, Both groups showed significantly decreased VAS, ODI, and Cobb angle 3 days after the operation and at the last follow-up when compared to values recorded pre-operation. However, we found no significant difference in VAS, ODI, and Cobb angle between the 2 groups (P > .05). We identified no complications (e.g., infection, VTE/PE, or nerve injury). Moreover, the pedicle screw placement position of the navigation group was better than that of the freehand group (P < .05), and the screw cortical penetration rate was lower than the freehand group (P < .05). During the process of posterior pedicle screw placement, O-arm navigation assisted by the Wiltse approach can significantly reduce operation time, minimize the amount of bleeding volume, and enhance the accuracy of pedicle screw implantation.

PML Body Component Sp100A Is a Cytosolic Responder to IFN and Activator of Antiviral ISGs.

Promyelocytic leukemia protein (PML) bodies are implicated in one of the key pathways in the establishment of antiviral status in response to interferon (IFN), yet the molecular mechanisms bridging the cross talk remain elusive. Herein, we report that a major constitutive component of the PML body, Sp100A, is ubiquitously located in the cytosol of various cell types and is an immediate responder to multiple extracellular stimuli, including virus infection, IFN, epidermal growth factor (EGF), glial cell-derived nerve factor (GDNF), etc., signaling through the phosphatidylinositol 3-kinase (PI3K) pathway. IFN-ß induces phosphorylation of Sp100A on Ser188, which fortifies the binding of Sp100A to pyruvate kinase 2 (PKM2) and facilitates its nuclear importation through the extracellular signal-regulated kinase 1/2 (ERK1/2)-PKM2-PIN1-importin axes. Blocking PI3K pathway signaling or interference with the ERK1/2-PKM2-PIN1-importin axes independently hampers nuclear translocation of Sp100A in response to IFN, reflecting a dual-regulation mechanism governing this event. In the nucleus, Sp100A is enriched in the promoter regions of essential antiviral interferon-stimulated genes (ISGs), such as those coding for IFI16, OAS2, and RIG-I, and activates their transcription. Importantly, nuclear importation of Sp100A, but not accumulation of a mutant Sp100A that failed to respond to IFN, during infection potently enhanced transcription of these antiviral ISGs and restricted virus propagation. These findings depict a novel IFN response mechanism by PML bodies in the cytosol and shed light on the complex sensing-regulatory network of PML bodies. IMPORTANCE PML bodies sit at the center stage of various important biological processes; however, the signal transduction networks of these macromolecular protein complexes remain enigmatic. The present study illustrates, in detail and for the first time, the course of signal receiving, processing, and implementation by PML bodies in response to IFN and virus infection. It shows that PML body constitutive component Sp100A was phosphorylated on Ser188 by IFN signaling through the PI3K pathway in the cytosol, cotranslocated into the nucleus with PKM2, enriched on the promoter regions of essential antiviral ISGs such as those coding for IFI16, RIG-I, OAS2, etc., and mediating their transcriptional activation.

Genomic characterization of two metagenome-assembled genomes of Tropheryma whipplei from China.

Whipple's disease is a rare chronic systemic disease that affects almost any organ system of the body caused by the intracellular bacterium Tropheryma whipplei, which is found ubiquitously in the environment. Sequencing of the T. whipplei genome has revealed that it has a reduced genome (0.93 Mbp), a characteristic shared with other intracellular bacteria. Until our research started, 19 T. whipplei strains had been sequenced from cultures originated in France, Canada, and Germany. The genome of T. whipplei bacterium has not been studied in Asia yet. Here, two metagenome-assembled genomes (MAGs) of T. whipplei from China were reconstructed through metagenomic next-generation sequencing (mNGS) and genome binning. We also provided genomic insights into the geographical role and genomic features by analyzing the whole genome. The whole-genome phylogenetic tree was constructed based on single-nucleotide polymorphism (SNP) distance calculations and then grouped by distance similarity. The phylogenetic tree shows inconsistencies with geographic origins, thus suggesting that the variations in geographical origins cannot explain the phylogenetic relationships among the 21 T. whipplei strains. The two Chinese strains were closely related to each other, and also found to be related to strains from Germany (T. whipplei TW08/27) and France (T. whipplei Bcu26 and T. whipplei Neuro1). Furthermore, the Average Nucleotide Identity (ANI) matrix also showed no association between geographic origins and genomic similarities. The pan-genome analysis revealed that T. whipplei has a closed pan-genome composed of big core-genomes and small accessory genomes, like other intracellular bacteria. By examining the genotypes of the sequenced strains, all 21 T. whipplei strains were found to be resistant to fluoroquinolones, due to the genetic mutations in genes gyrA, gyrB, parC, and parE. The 21 T. Whipplei strains shared the same virulence factors, except for the alpC gene, which existed in 7 out of the 21 T. whipplei strains. When comparing 21 entire T. whipplei pan-genomes from various nations, it was discovered that the bacterium also possessed a closed genome, which was a trait shared by intracellular pathogens.

Efficient regeneration of mature castanopsis hystrix from in vitro stem explants.

Castanopsis hystrix is one of the main timber trees grown in China. However, severe shortage of natural seeds and the difficulty of explant regeneration has limited seedling supply. As such, there is a need for research on asexual multiplication of C. hystrix. This study established a rapid propagation technology system for C. hystrix genotypes, including explant treatment, proliferation, and rooting. HZ (a modified MS medium) supplemented with 4.4 µM BA and 0.5 µM IBA was found to be the optimal medium for shoot sprouting. The maximum proliferation coefficient and the number of effective shoots was obtained on HZ medium supplemented with 2.6 µM BA and 1.0 µM IBA, were 3.00 and 5.63, respectively. A rooting rate of 83.33% was achieved using half-strength HZ medium supplemented with 3.2 µM NAA. Adding vitamin C (80 mg⋅l-1) for 7 days in a dark environment reduced the browning rate, while increasing the proliferation rate. Additionally, through cytological observation, we established how and where adventitious roots occur. The survival rate of transplanted plantlets was > 90%. This is the first report of an in vitro regeneration technique that uses stem segments of mature C. hystrix as explants.

Effect of body weight on bleeding events of aspirin in ischemic stroke or transient ischemic attack patients.

WHAT IS KNOWN AND OBJECTIVES: Bleeding is the most common adverse reaction to aspirin and can lead to drug discontinuation or even be life-threatening in the secondary prevention of stroke or transient ischemic attack. The aim of this study was to evaluate risk factors for bleeding adverse reaction of aspirin in ischemic stroke or transient ischemic attack. METHODS: This retrospective analysis included patients treated with aspirin (100 mg) as a secondary prevention for ischemic stroke or transient ischemic attack. The bleeding events that occurred during the first year were collected, including gastrointestinal, skin, nasal cavity, gum, and urinary tract bleeding events. Then, univariate and multivariate logistic regression analyses were used to identify independent factors associated with bleeding events of aspirin. RESULTS AND DISCUSSION: A total of 578 patients were enrolled in this study, and 58 patients developed bleeding during the first year of secondary prevention. Body weight and combination with selective serotonin reuptake inhibitors were found to be significant risk factors for overall bleeding (p = 0.025 and 0.012). Body weight below 60 kg was a risk factor for overall bleeding and gastrointestinal bleeding events. WHAT IS NEW AND CONCLUSION: Patients weighing less than 60 kg were at increased risk of bleeding with 100 mg aspirin for secondary prevention of cerebral infarction transient ischemic attack.

Sugar Metabolism and Transcriptome Analysis Reveal Key Sugar Transporters during Camellia oleifera Fruit Development.

Camellia oleifera is a widely planted woody oil crop with economic significance because it does not occupy cultivated land. The sugar-derived acetyl-CoA is the basic building block in fatty acid synthesis and oil synthesis in C. oleifera fruit; however, sugar metabolism in this species is uncharacterized. Herein, the changes in sugar content and metabolic enzyme activity and the transcriptomic changes during C. oleifera fruit development were determined in four developmental stages (CR6: young fruit formation; CR7: expansion; CR9: oil transformation; CR10: ripening). CR7 was the key period of sugar metabolism since it had the highest amount of soluble sugar, sucrose, and glucose with a high expression of genes related to sugar transport (four sucrose transporters (SUTs) or and one SWEET-like gene, also known as a sugar, will eventually be exported transporters) and metabolism. The significant positive correlation between their expression and sucrose content suggests that they may be the key genes responsible for sucrose transport and content maintenance. Significantly differentially expressed genes enriched in the starch and sucrose metabolism pathway were observed in the CR6 versus CR10 stages according to KEGG annotation. The 26 enriched candidate genes related to sucrose metabolism provide a molecular basis for further sugar metabolism studies in C. oleifera fruit.

In vitro Studies and Clinical Observations Imply a Synergistic Effect Between Epstein-Barr Virus and Dengue Virus Infection.

Dengue virus (DENV) infection can lead to a complex spectrum of clinical outcomes, ranging from asymptomatic infection to life-threatening severe dengue. The reasons for thus drastically varying manifestations of the disease remain an enigma. Herein, we reported an original discovery of the synergistic effect between preexisting Epstein-Barr virus (EBV) infection and DENV superinfection in vitro and of a strong correlation of these two viruses in the clinical samples from dengue patients. We showed that (I) DENV-2 infection of an EBV-positive cell line (EBV + Akata cell) reactivated EBV, and it could be blocked by wortmannin treatment. (II) Examination of human peripheral blood mononuclear cell (PBMC) samples from dengue patients revealed significantly elevated cell-associated EBV DNA copy number at the time of hospitalization vs. at the time of disease recovery in most individuals. (III) EBV infection promoted DENV propagation in both EBV-hosting B cells and indirectly in THP-1 cells, supported by the following evidence: (A) EBV + Akata cells were more permissive to DENV-2 infection compared with Akata cells harboring no EBV virus (EBV- Akata cells). (B) Low-molecular weight fraction secreted from EBV + Akata cells could enhance DENV-2 propagation in monocytic THP-1 cells. (C) While reactivation of EBV in EBV + Akata cells further increased DENV-2 yield from this cell line, pharmacological inhibition of EBV replication by acyclovir had the opposite effect. To our knowledge, this is the first investigation demonstrating a positive correlation between EBV and DENV in vitro and in human biospecimens.

Efficient Tissue Culture Protocol for Magnolia lucida (Magnoliaceae) and Confirmation of Genetic Stability of the Regenerated Plants.

Magnolia lucida (Magnoliaceae) is classified as an endangered species by the International Union for Conservation of Nature. It has high commercial value owing to its attractive tree shape and flowers. We adopted an excellent genotype of M. lucida as the parent material and established a mini-cut orchard through grafting to provide trunk shoots explants over the long-term. Optimal sterilization was achieved using a combination of 75% ethanol for 30 s, one percent benzalkonium bromide for five minutes, and 0.1% mercuric chloride for five minutes. Modified Murashige and Skoog medium (ML) was the optimal medium for the growth of M. lucida. Addition of one mg/L of 6-benzyl adenine (BA) and 0.05 mg/L of α-naphthaleneacetic acid (NAA) to the medium increased the shoot induction rate to 95.56%, and the ML medium containing 0.4 mg/L BA and 0.04 mg/L NAA achieved the maximum multiplication rate (284.56%). Dark treatment for seven days, followed by continuous light treatment could better resolve the challenge of difficult rooting in M. lucida plants. Using random amplified polymorphic DNA and inter simple sequence repeat markers, we confirmed the genetic uniformity and stability of the regenerated plants. Our protocol should be helpful for the propagation and conservation of this endangered plant.

A risky output of variation orders in renewable energy projects: Identification, assessment and validation.

Renewable energy projects (REPs) are constructed to contribute to the eradication of global warming and supporting a sustainable aspect of economies. However, due to their entrepreneurial nature, such REPs are quite prone to severe project variations. Mostly, REPs deviate from their allocated plans, which lead them to the time and cost overrun outcomes alongside numerous other hindering impacts. The current research was intended to understand the critical impacts of project variations during the building activities of REPs. Utilizing a sample of 253 field experts engaged with such REPs at Pakistan, we employed structural equation modeling (SEM) to analyze and confirm our propositions. It was found that cost-related effects were the most significant outcomes in the REPs and their severity was higher than other variation consequences of time-overrun effects, productivity-related effects, risk-related effects, and reputation related effects. Nevertheless, an unexpected outcome was observed regarding the variations, which do not show their negative effects on reputation related aspects in the REPs of Pakistan. Considering its critical findings, this research is a useful tool for project executives that they can formulate certain techniques to tackle such variations and their consequences in REPs. This research likewise aimed to add and extend pertinent literature for REPs by following the outcomes of the current research concerning the impacts of project variations.

An efficient micropropagation protocol for an endangered ornamental tree species (Magnolia sirindhorniae Noot. & Chalermglin) and assessment of genetic uniformity through DNA markers.

Magnolia sirindhorniae Noot. & Chalermglin is an endangered species with high ornamental and commercial value that needs to be urgently protected and judiciously commercialized. In this study, a protocol for efficient regeneration of this species is standardized. The lateral buds of the M. sirindhorniae plant were used as an explant. Half-strength Murashige and Skoog (MS) medium supplemented with 2.0 mg/L 6-benzyladenine (BA), 0.1 mg/L α-naphthaleneacetic acid (NAA), and 2.0 mg/L gibberellic acid (GA3) was found to be the optimal medium for shoot induction. The maximum shoot multiplication rate (310%) was obtained on Douglas-fir cotyledon revised medium (DCR) fortified with 0.2 mg/L BA, 0.01 mg/L NAA, and additives. The half-strength DCR medium supplemented with 0.5 mg/L NAA and 0.5 mg/L indole-3-butyric acid (IBA) supported the maximum rate (85.0%) of in vitro root induction. After a simple acclimatization process, the survival rate of plantlets in a substrate mixture of sterile perlite and peat soil (1:3; v/v) was 90.2%. DNA markers were used for assessment of genetic uniformity, confirming the genetic uniformity and stability of regenerated plants of M. sirindhorniae. Thus, the described protocol can safely be applied for large scale propagation of this imperative plant.

Manglietia pubipedunculata (Magnoliaceae), a new species from Yunnan, China.

A new species, Manglietia pubipedunculata Q. W. Zeng & X. M. Hu (Magnoliaceae) is described and illustrated from Yunnan, China. In addition to macromorphological examination, we comparatively studied on micromorphology of leaf epidermis, leaf structure, and epidermal cell on the sclerotesta. This new species is similar to M. kwangtungensis in terms of having dense pubescence, however, their pubescence are quite different. Manglietia pubipedunculata has appressed, compressed, shorter and sparser pubescence consisting of single or two cells. Moreover, it differs from M. kwangtungensis by showing shorter and thicker peduncles, longer styles, basal carpels covered with sparsely brown appressed pubescence, and more ovules per carpel. Furthermore, the new species has thinner leaves, brown and rugged surfaces on sclerotesta, and the alveolate cell pattern consisting of pentagon or hexagon cells with papilla on secondary cell wall under the observation by SEM. The phylogenetic analysis from two nuclear PHYA and LEAFY and chloroplast trnH-psbA sequences of 11 taxa reveals that M. pubipedunculata is a distinct species.

Evolution of interspecies unilateral incompatibility in the relatives of Arabidopsis thaliana.

The evolutionary concurrence of intraspecies self-incompatibility (SI) and explosive angiosperm radiation in the Cretaceous have led to the hypothesis that SI was one of the predominant drivers of rapid speciation in angiosperms. Interspecies unilateral incompatibility (UI) usually occurs when pollen from a self-compatible (SC) species is rejected by the pistils of a SI species, while the reciprocal pollination is compatible (UC). Although this SI × SC type UI is most prevalent and viewed as a prezygotic isolation barrier to promote incipient speciation of angiosperms, comparative evidence to support such a role is lacking. We show that SI × SI type UI in SI species pairs is also common in the well-characterized accessions representing the four major lineages of the Arabidopsis genus and is developmentally regulated. This allowed us to reveal a strong correlation between UI strength and species divergence in these representative accessions. In addition, analyses of a SC accession and the pseudo-self-compatible (psc) spontaneous mutant of Arabidopsis lyrata indicate that UI shares, at least, common pollen rejection pathway with SI. Furthermore, genetic and genomic analyses of SI × SI type UI in A. lyrata × A. arenosa species pair showed that two major-effect quantitative trait loci are the stigma and pollen-side determinant of UI, respectively, which could be involved in heterospecies pollen discrimination. By revealing a close link between UI and SI pathway, particularly between UI and species divergence in these representative accessions, our findings establish a connection between SI and speciation. Thus, the pre-existence of SI system would have facilitated the evolution of UI and accordingly promote speciation.

Anticancer effect of exogenous hydrogen sulfide in cisplatin­resistant A549/DDP cells.

Despite huge advances in lung cancer treatment, resistance to cisplatin­based chemotherapy remains one of the major obstacles, and the elucidation of cisplatin resistance remains challenging. As an important biological and pharmacological mediator, hydrogen sulfide (H2S) performs a variety of homeostatic functions related to cancer formation and development. However, the effects of H2S on cisplatin­resistance lung cancer remain largely unknown. In the present study, we investigated the anticancer effects and relevant mechanisms of NaHS (an exogenous donor of H2S) on A549/DDP cells (cisplatin­resistant). The intracellular H2S was first evaluated using a fluorescence probe in A549 (cisplatin­sensitive) and A549/DDP cells. We found that H2S production was markedly decreased in A549/DDP cells compared with that in A549 cells, accomplished by the downregulation of cystathionine ß­synthase (CBS), an endogenous H2S­producing enzyme. In view of these findings, we then observed the effects of NaHS treatment on A549/DDP cells. The results showed that NaHS exposure exhibited an inhibitory effect on cell viability and the IC50 of cisplatin in A549/DDP cells decreased markedly during NaHS treatment (800 µmol/l). In addition, our data revealed that NaHS treatment of A549/DDP cells resulted in the induction of apoptosis, cell cycle arrest and inhibition of cell migration and invasion. Finally, we demonstrated that the marked changes in the A549/DDP cell response to NaHS may be triggered by the activation of p53, and overexpression of p21, caspase­3, Bax and MMP­2, as well as the downregulation of Bcl­xL. The findings of the present study provide novel evidence that NaHS administration may represent a new strategy for the treatment of cisplatin­resistant lung cancer.

Phylogeographic pattern suggests a general northeastward dispersal in the distribution of Machilus pauhoi in South China.

Machilus pauhoi Kanehira is an important timber species in China. A provenance trial was recently set up to evaluate the growth performance of trees from different localities, with the aim of designing seed transfer guidelines. Here, we tested twelve nuclear microsatellite markers derived from other species of the Lauraceae family and investigated population genetic structure in M. pauhoi. Both the number of observed alleles per locus (Na) and the polymorphic information content (PIC) significantly decreased against the latitude, but showed an insignificant decrease against the longitude. Heterozygosity (Ho) and gene diversity (h) exhibited a weak correlation with geographic location. Private alleles were present in multiple populations, and a moderate level of population genetic differentiation was detected (Gst = 0.1691). The joint pattern of genetic diversity (Na, PIC, Ho, and h) suggests that general northeastward dispersal led to the current distribution of M. pauhoi. Significant but weak effects of isolation-by-distance (IBD) occurred, implicating the mountain ranges as the major barrier to gene flow. Both STRUCTURE and hierarchical clustering analyses showed three distinct groups of populations related to the physical connectivity among mountain ranges. A priority in designing genetic conservation should be given to the populations at the southwest side of the species' distribution. This conservation strategy can also be combined with the pattern of adaptive genetic variation from the provenance trial for comprehensive genetic resource management of native M. pauhoi.

Serum is an indispensable factor in the maintenance of the biological characteristics of sweat gland cells.

The tolerance of sweat gland cells for in vitro amplification and subcultivation is low as they are somatic cells. The present study aimed to formulate an optimal medium for the culture of human eccrine sweat gland cells (HESGCs) and to establish a method for induction of HESGCs proliferation, whilst maintaining the characteristics of sweat gland cells. HESGCs cultured in sweat gland (SG):keratinocyte growth medium­2 (KGM­2) (1:1) medium had a higher proliferation rate and a stable morphology compared with cells cultured in SG and KGM­2 medium only. Reverse transcription­quantitative polymerase chain reaction indicated that cells cultured in the SG:KGM­2 (1:1) medium exhibited higher expression levels of α­smooth muscle actin, keratin (K)77, carcinoembryonic antigen, K8, K18, ectodysplasin A receptor, c­Myc, Kruppel­like factor 4 and octamer­binding transcription factor 4 compared with cells cultured in SG only or KGM­2 only medium. Three­dimensional culture analysis revealed that HESGCs cultured in SG:KGM­2 1:1 medium differentiated into sweat gland­like structures, whereas cells cultured in KGM­2 only medium underwent cornification. The present study also determined that the maintenance of the biological characteristics of HESGCs occurred due to the presence of fetal bovine serum (FBS). Cells cultured in medium without FBS differentiated into keratinocytes. Therefore, the SG:KGM­2 (1:1) medium may be a suitable culture medium for HESGCs. In conclusion, this mixed medium is a valuable compound and should be considered to be a potential supplemental medium for HESGCs.

Characterization of hemicelluloses from Neolamarckia cadamba (Rubiaceae) during xylogenesis.

In this study, hemicelluloses were isolated from the apical, middle and basal segments of N. cadamba using a 2N KOH extraction procedure. Chemical composition and structural characterization of the three hemicellulosic fractions obtained were comparatively investigated by a combination of HPLC, GPC, FTIR, 1H,13C, HSQC NMR and TGA techniques. According to the sugar analysis and spectral results, (4-O-methyl) glucuronoxylan was the primary hemicellulose identified in the samples with trace levels of mannan also present. All of the three samples showed spherical polymers in 0.005M sodium phosphate solution (containing 0.02N NaCl). Xylan in the middle and basal segment stems had higher molecular weights. Our findings show that during xylogenesis in N. cadamba, changes are observed in xylan content and molecular weight.