The metabolic slowdown caused by the deletion of pspA accelerates protein aggregation during stationary phase facilitating antibiotic persistence.

Entering a dormant state is a prevailing mechanism used by bacterial cells to transiently evade antibiotic attacks and become persisters. The dynamic progression of bacterial dormancy depths driven by protein aggregation has been found to be critical for antibiotic persistence in recent years. However, our current understanding of the endogenous genes that affects dormancy depth remains limited. Here, we discovered a novel role of phage shock protein A (pspA) gene in modulating bacterial dormancy depth. Deletion of pspA of Escherichia coli resulted in increased bacterial dormancy depths and prolonged lag times for resuscitation during the stationary phase. ∆pspA exhibited a higher persister ratio compared to the wild type when challenged with various antibiotics. Microscopic images revealed that ∆pspA showed accelerated formation of protein aggresomes, which were collections of endogenous protein aggregates. Time-lapse imaging established the positive correlation between protein aggregation and antibiotic persistence of ∆pspA at the single-cell level. To investigate the molecular mechanism underlying accelerated protein aggregation, we performed transcriptome profiling and found the increased abundance of chaperons and a general metabolic slowdown in the absence of pspA. Consistent with the transcriptomic results, the ∆pspA strain showed a decreased cellular ATP level, which could be rescued by glucose supplementation. Then, we verified that replenishment of cellular ATP levels by adding glucose could inhibit protein aggregation and reduce persister formation in ∆pspA. This study highlights the novel role of pspA in maintaining proteostasis, regulating dormancy depth, and affecting antibiotic persistence during stationary phase.

Emergence of Antifungal Resistant Subclades in the Global Predominant Phylogenetic Population of Candida albicans.

Candida albicans remains the most common species causing invasive candidiasis. In this study, we present the population structure of 551 global C. albicans strains. Of these, the antifungal susceptibilities of 370 strains were tested. Specifically, 66.6% of the azole-nonsusceptible (NS)/non-wild-type (NWT) strains that were tested belonged to Clade 1. A phylogenetic analysis, a principal components analysis, the population structure, and a loss of heterozygosity events revealed two nested subclades in Clade 1, namely, Clade 1-R and Clade 1-R-α, that exhibited higher azole-NS/NWT rates (75.0% and 100%, respectively). In contrast, 6.4% (21/326) of the non-Clade 1-R isolates were NS/NWT to at least 1 of 4 azoles. Notably, all of the Clade 1-R-α isolates were pan-azole-NS/NWT that carried unique A114S and Y257H double substitutions in Erg11p and had the overexpression of ABC-type efflux pumps introduced by the substitution A736V in transcript factor Tac1p. It is worth noting that the Clade 1-R and Clade 1-R-α isolates were from different cities that are distributed over a large geographic span. Our study demonstrated the presence of specific phylogenetic subclades that are associated with antifungal resistance among C. albicans Clade 1, which calls for public attention on the monitoring of the future spread of these clones. IMPORTANCE Invasive candidiasis is the most common human fungal disease among hospitalized patients, and Candida albicans is the predominant pathogen. Considering the large number of infected cases and the limited alternative therapies, the azole-resistance of C. albicans brings a huge clinical threat. Here, our study suggested that antifungal resistance in C. albicans could also be associated with phylogenetic lineages. Specifically, it was revealed that more than half of the azole-resistant C. albicans strains belonged to the same clade. Furthermore, two nested subclades of the clade exhibited extremely high azole-resistance. It is worth noting that the isolates of two subclades were from different cities that are distributed over a large geographic span in China. This indicates that the azole-resistant C. albicans subclades may develop into serious public health concerns.

Differential efficacy of water lily cultivars in phytoremediation of eutrophic water contaminated with phosphorus and nitrogen.

Excessive inputs of phosphorus (P) and nitrogen (N) trigger eutrophication of the water bodies, which promotes the undesirable growth of algal bloom and deterioration of the water quality, and aquatic biodiversity. Macrophytes provide an environmentally benign and economically viable paradigm for the ecological restoration of eutrophic aquatic ecosystems. Water lily (Nymphaea) is largely used as ornamental plant for landscaping, and it has been documented that water lily possesses the potentiality in mitigating polluted aquatic environments. In the present study, water lily cultivars Nymphaea Texas Dawn (NTD), Nymphaea Colorado (NC), Nymphaea Madame Wilfron Gonnère (NMWG), and Nymphaea Sunshine Princess (NSP) were investigated for their potency in alleviating the eutrophication. The concentrations of total P and total N were significantly higher in the leaves of NC and NSP compared with NTD and NMWG. Therefore, NC and NSP were selected for subsequent studies to decipher their recuperation efficacy on eutrophic waters at different growth stages. NC and NSP significantly reduced the concentrations of eutrophication indicators i.e., total P, NH4+-N, and chemical oxygen demand in different gradients of the simulated eutrophic water in a growth-dependent manner. On the contrary, NC and NSP triggered a significant increase in the concentration of dissolved oxygen particularly at the seedling stage. Notably, the concentrations of total P (shoot and root) and total N (root) were relatively higher in NSP than NC. The study thus revealed a growth-dependent differential efficacy of NSP and NC in mitigating the different eutrophic waters.

Effects of ethylene biosynthesis and signaling on oxidative stress and antioxidant defense system in Nelumbo nucifera G. under cadmium exposure.

Water contamination with cadmium (Cd) is a global environmental problem and its remediation becomes urgent. Phytoremediation using ornamental plants has attracted much attention for its advantages of cost-effectiveness and beautification of the environment. Nelumbo nucifera G. is a popular ornamental aquatic macrophyte with fast growth, large biomass, and high capacities for Cd accumulation and removal. However, information about Cd resistance and defense responses in N. nucifera is rather scarce, which restricts its large-scale utilization for phytoremediation. The phytohormone ethylene plays an important role in plant resistance to Cd stress, but the underlying mechanism remains unclear. In this study, we investigated morphophysiological responses of N. nucifera seedlings to Cd stress, and focused on the effects of ethylene on oxidative damage, Cd accumulation, and antioxidant defense system at the metabolic and transcript levels in leaves under Cd stress. Our results showed that Cd exposure led to leaf chlorosis and necrosis, coupled with an increase in contents of hydrogen peroxide, electrolyte leakage, and malondialdehyde, and decrease in chlorophyll content. Exogenous ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) application intensified Cd-induced stress responses and Cd accumulation, and increased ethylene production by inducing ACC synthase (ACS) gene NnACS. Such enhanced ethylene emission inhibited catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) activities, and modulated ascorbate (AsA) and glutathione (GSH) accumulation through transcriptional regulation of their respective metabolic genes. After ethylene action inhibitor silver thiosulfate (STS) supplementation, Cd-induced oxidative damage was abolished, and Cd content declined but still at a relatively high level. Blocking of ethylene perception by STS inhibited ethylene biosynthesis; enhanced CAT, APX, and GR activities and their transcript levels; increased AsA accumulation via inducing its biosynthetic genes; but reduced GSH content and NnGSH2 expression level. These results suggest that ethylene biosynthesis and signaling play an important role in N. nucifera response to Cd stress, and maintaining appropriate ethylene level and low ethylene sensitivity could improve its Cd tolerance via efficient antioxidant defenses.

Successful treatment of plantar warts using topical Zijinding, a traditional Chinese medicine preparation: A case series.

INTRODUCTION: Plantar warts are associated with high transmissibility and morbidity. Among the available therapeutic options, none is uniformly effective or virucidal. Salicylic acid is the first-line therapy but approximately one-third of lesions could not resolve and become recalcitrant despite repeated treatment. Cryotherapy is widely accessible with low cost but may be complicated by pain, blister formation, hemorrhage, infection, excessive granulation tissue formation, and hyper-/hypo-pigmentation. Hence, alternative treatment modalities are essential. METHODS: Three patients with debilitating plantar warts refractory or intolerant to cryotherapy were treated with a course of Zijinding (a traditional Chinese medicine preparation) paste prepared with white vinegar. RESULTS: All three patients showed excellent clinical response with Zijinding application with evolution of lesions to scabs and subsequently healthy skin within 1.5 to 5 months of treatment. Treatment was well tolerated and had no significant side effects with excellent compliance recorded for all three patients. There was no relapse for at least 10 months after stopping the treatment. CONCLUSION: Topical Zijinding could be a promising alternative modality for the treatment of plantar warts. Further clinical trials on the comparison of Zijinding and other treatment modalities of plantar warts are warranted. Further studies are required to investigate the mechanism of action of Zijinding and to isolate the active ingredient.

Traditional Chinese medicine Dingkun Pill facilitates uterine receptivity for implantation in mice†.

Dingkun Pill (DK) is one of the representative traditional Chinese medicines, which has been used in the treatment of gynecological diseases for hundreds of years. Accumulative observations and evidence have shown the beneficial effects of DK, including enhancing the function of hypothalamic-pituitary-ovarian axis. However, the underlying mechanisms remain elusive. In this study, the effects of DK on uterine receptivity and implantation were explored by a series of studies with different mouse models. The results showed that DK can advance the time of implantation by influencing the expression of estrogen target genes to facilitate embryo implantation. DK was efficient to activate embryo implantation at the presence of suboptimal estrogen in delayed implantation mouse model. Our further study revealed that the improvement of DK on receptivity establishment is attributed to the differential regulation of DK on implantation-associated genes. This study provides previously unappreciated molecular mechanism of DK in embryo implantation and benefits the potential clinical application of DK in human reproduction improvement.

Genome-wide analysis of the Solanum tuberosum (potato) trehalose-6-phosphate synthase (TPS) gene family: evolution and differential expression during development and stress.

BACKGROUND: Trehalose-6-phosphate synthase (TPS) serves important functions in plant desiccation tolerance and response to environmental stimuli. At present, a comprehensive analysis, i.e. functional classification, molecular evolution, and expression patterns of this gene family are still lacking in Solanum tuberosum (potato). RESULTS: In this study, a comprehensive analysis of the TPS gene family was conducted in potato. A total of eight putative potato TPS genes (StTPSs) were identified by searching the latest potato genome sequence. The amino acid identity among eight StTPSs varied from 59.91 to 89.54%. Analysis of dN/dS ratios suggested that regions in the TPP (trehalose-6-phosphate phosphatase) domains evolved faster than the TPS domains. Although the sequence of the eight StTPSs showed high similarity (2571-2796 bp), their gene length is highly differentiated (3189-8406 bp). Many of the regulatory elements possibly related to phytohormones, abiotic stress and development were identified in different TPS genes. Based on the phylogenetic tree constructed using TPS genes of potato, and four other Solanaceae plants, TPS genes could be categorized into 6 distinct groups. Analysis revealed that purifying selection most likely played a major role during the evolution of this family. Amino acid changes detected in specific branches of the phylogenetic tree suggests relaxed constraints might have contributed to functional divergence among groups. Moreover, StTPSs were found to exhibit tissue and treatment specific expression patterns upon analysis of transcriptome data, and performing qRT-PCR. CONCLUSIONS: This study provides a reference for genome-wide identification of the potato TPS gene family and sets a framework for further functional studies of this important gene family in development and stress response.

Epidemiology of candidemia and antifungal susceptibility in invasive Candida species in the Asia-Pacific region.

In the Asia-Pacific region, Candida albicans is the predominant Candida species causing invasive candidiasis/candidemia in Australia, Japan, Korea, Hong Kong, Malaysia, Singapore and Thailand whereas C. tropicalis is the most frequently encountered Candida species in Pakistan and India. Invasive isolates of C. albicans, C. parapsilosis complex and C. tropicalis remain highly susceptible to fluconazole (>90% susceptible). Fluconazole resistance (6.8-15%), isolates with the non-wild-type phenotype for itraconazole susceptibility (3.9-10%) and voriconazole (5-17.8%), and echinocandin resistance (2.1-2.2% in anidulafungin and 2.2% in micafungin) among invasive C. glabrata complex isolates are increasing in prevalence. Moreover, not all isolates of C. tropicalis have been shown to be susceptible to fluconazole (nonsusceptible rate, 5.7-11.6% in China) or voriconazole (nonsusceptible rate, 5.7-9.6% in China).

De novo transcriptome sequencing and discovery of genes related to copper tolerance in Paeonia ostii.

Paeonia ostii, an important ornamental and medicinal plant, grows normally on copper (Cu) mines with widespread Cu contamination of soils, and it has the ability to lower Cu contents in the Cu-contaminated soils. However, very little molecular information concerned with Cu resistance of P. ostii is available. In this study, high-throughput de novo transcriptome sequencing was carried out for P. ostii with and without Cu treatment using Illumina HiSeq 2000 platform. A total of 77,704 All-unigenes were obtained with a mean length of 710 bp. Of these unigenes, 47,461 were annotated with public databases based on sequence similarities. Comparative transcript profiling allowed the discovery of 4324 differentially expressed genes (DEGs), with 2207 up-regulated and 2117 down-regulated unigenes in Cu-treated library as compared to the control counterpart. Based on these DEGs, Gene Ontology (GO) enrichment analysis indicated Cu stress-relevant terms, such as 'membrane' and 'antioxidant activity'. Meanwhile, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis uncovered some important pathways, including 'biosynthesis of secondary metabolites' and 'metabolic pathways'. In addition, expression patterns of 12 selected DEGs derived from quantitative real-time polymerase chain reaction (qRT-PCR) were consistent with their transcript abundance changes obtained by transcriptomic analyses, suggesting that all the 12 genes were authentically involved in Cu tolerance in P. ostii. This is the first report to identify genes related to Cu stress responses in P. ostii, which could offer valuable information on the molecular mechanisms of Cu resistance, and provide a basis for further genomics research on this and related ornamental species for phytoremediation.

Clinical Characteristics, Laboratory Identification, and In Vitro Antifungal Susceptibility of Yarrowia (Candida) lipolytica Isolates Causing Fungemia: a Multicenter, Prospective Surveillance Study.

Our case series showed that uncomplicated Yarrowia lipolytica fungemia might be treated with catheter removal alone. The Vitek 2 YST identification (ID) card system, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), and internal transcribed spacer and 25S nuclear ribosomal DNA (nrDNA) gene sequencing provided reliable identification. All isolates had low MICs to voriconazole, echinocandins, and amphotericin B.

Development of fluconazole resistance in a series of Candida parapsilosis isolates from a persistent candidemia patient with prolonged antifungal therapy.

BACKGROUND: Candida parapsilosis was the most common species causing candidemia in the 2010 China Hospital Invasive Fungal Surveillance Net (CHIF-NET) database. Compared to Candida albicans, the description of azole resistance and mechanisms in C. parapsilosis is very limited. We report a patient with C. parapsilosis candidemia over several months, due to a probable intravascular source, who developed fluconazole resistance after prolonged treatment. CASE PRESENTATION: An 82 year-old male had a hospital admission of approximately 1.5 years duration. He was initially admitted with acute pancreatitis. Prior to succumbing to the illness, he developed candidemia and treated with three antifungal drugs for nearly 5 months, at suboptimal doses and without source control. Following treatment, 6 blood cultures were still positive for C. parapsilosis. The last 2 strains were resistant to fluconazole (MICs 32 µg/mL) and intermediate to voriconazole (MICs 0.5 µg/mL). Microsatellite multilocus analysis indicated that the 6 isolates from the patient belonged to a single genotype. The first 4 isolates were susceptible to fluconazole (MICs 2 µg/mL) and voriconazole (MICs 0.015-0.03 µg/mL), which were slightly higher than susceptible control strains from other patients. Overexpression of MDR1 genes were detected in the two resistant isolates, and this was associated with a homozygous mutation in MRR1 genes (T2957C /T2957C), with the amino acid exchange L986P. CONCLUSIONS: This case corroborates that the resistant C. parapsilosis isolates can emerge in the setting of complicated infections and the extensive use of antifungal agents, emphasizing the need for standardizing and improving the antifungal treatment as well as source control in the treatment of infection diseases.

A systematic review of vinorelbine for the treatment of breast cancer.

The purpose of this study was to investigate the efficacy and safety profile of vinorelbine-based chemotherapy in different settings for the treatment of breast cancer. We performed a computerized search using combinations of the following keywords: "breast cancer", "breast neoplasms", "trial", "vinorelbine" and "navelbine". A total of 20 trials were included in this analysis, with a total of 5,080 patients accrued. Taxane was associated with enhanced overall survival (OS; p = 0.027) and response rate (RR; p = 0.037) as compared with vinorelbine in monotherapy, but did not show significantly favored progression-free survival (PFS; p = 0.136). Vinorelbine alone was equivalent to fluoropyrimidine treatment in RR (p = 0.79) for the treatment of metastatic breast cancer. For vinorelbine-combined regimens, the analysis showed that the vinorelbine group gave similar results as other regimens for OS (p = 0.849) and PFS (p = 0.143). The RR of vinorelbine-combined regimens was slightly better than that of the other regimens (OR, 1.17), but the difference was not statistically significant. In neoadjuvant setting, vinorelbine treatment was as active as AC (doxorubicin, cyclophosphamide) or DAC (doxorubicin, cyclophosphamide, docetaxel) regimens with respect to RR (p = 0.76) and pathologic complete response (pCR; p = 0.77), but showed lower occurrence of grade 3/4 adverse effects. The analysis also demonstrated that vinorelbine-containing therapy is effective as adjuvant, front-line or salvage therapy of metastatic breast cancer, even for patients who were previously treated with anthracyclines or taxanes.

[Effects of water stress on the growth of Lonicera japonica and quality of honeysuckle].

OBJECTIVE: Effects of water stress on the growth of Lonicera japonica and quality of honeysuckle were studied. METHODS: Different extent water stress treatment imposed to potted plants before the first florescence of honeysuckle, and the growth and related physiological indexes of Lonicera japonica were determined. RESULTS: Shoot grow slowly as well as the increases of specific weight of leaf and the decrease of the yield of honeysuckle under water stress. The contents of chlorophyl in the leaves treated water stress all increased, and the content of soluble suger had a rising stream. The content of soluble protein in leaves of plant imposed by minor stress treatment was obviously higher than that of CK and severe stress treatment. As the stress was hard, the content of proline in leaves increased significantly. With water stress severing, the content of chlorogenic acid in the flower bud decreased very significantly than CK. CONCLUSION: Water condition was a important factor affected the growth and development of shoots, leaves and flower buds as well as the quality of honeysuckle. In the cultivation of Loniccra japonica, irrigation timing and certain irrigation condition was necessary.

Study on atherosclerosis treated with theory of detoxification.

Starting with the contents, classification and pathogenic characteristics of the toxic pathogen and combining the modern medical research on the correlation of atherosclerosis with inflammation and immune reaction, authors have studied and expounded the interrelationship between the toxic pathogen and atherosclerosis. The toxic pathogen affecting the whole pathological process of atherosclerosis is a key factor for the disease to remain lingering and a cause of various cardiocerebrovascular diseases. Detoxification can be used to treat atherosclerosis so as to enhance the toxin-removing ability of the body and resist the damage to the body from the toxic pathogen.

[Effects of irrigation volume on growth and quality of Lonicera japonica].

OBJECTIVE: To study the effects of irrigation volume on the growth and quality of Lonicera japonica. METHOD: Different volume of irrigation water was applied to the potted L. japonica before the first florescence of honeysuckle in order to keep the relative moisture content of the soil as 100%, 80%, 50%, 30%, and observe the growth and development of shoot and leaves, yield and quality of honeysuckle. RESULT: As the of irrigation volume was reduced, the internodal elongation of shoot and the increase of the knot were restrained, and the yield of honeysuckle dropped. But the content of chlorogen acid in the buds treated by 80% irrigation volume was not affected, while that of the other treatment all decreased. The moisture contents of the leaves treated by 30% irrigation volume was significantly lower than that of the CK, while the specific leaf weight (SLW) increased significantly comparing to the CK. The content of chlorophyl in the leaves of 50% and 30% increased, while the ratio of chlorophyl A and B fell significantly. The content of dissoluble suger in the leaves of 80% and 50% irrigation volume was much higher than that in the CK. The results also showed that, the content of dissoluble protein in the leaves of 30% irrigation volume increased comparing with that in 50%, and the content of proline in leaveas of 30% irrigation volume increased significantly. CONCLUSION: Using less irrigation volume (keeping 80% of relative moisture content of the soil) does not affect the quality of honeysuckle, but decreased dry weight of honeysuckle. It is necessary to take irrigation management during cultivation of honeysuckle.

[Clinical study on drug-seperated moxibustion at Shenque (CV 8) for treatment of ulcerative colitis].

OBJECTIVE: To observe the therapeutic effect of drug-separated moxibustion at Shenque (CV 8) on ulcerative colitis (UC) and the influence on autoimmunity level, and study on the mechanism. METHODS: Sixty cases were randomly divided into a treatment group and a control group, 30 cases in each group. The treatment group were treated with drug-separated moxibustion at Shenque (CV 8) and the control group with oral administration of sulphasalazine and metronidazole tablets. The main symptoms, pathological changes of the intestinal mucosa and the total therapeutic effect in the two groups before and after treatment, and the changes of the content of blood serum immunoglobulin, peripheral blood T-cell subgroup and NK cell were observed. RESULTS: The markedly effective rate and the total effective rate in the treatment group were 60.0% and 86.7% respectively, much higher than the control group (P<0.05, P<0.01). IgG content in the treatment group after treatment significantly decreased with a significant difference as compared with the control group (P<0.05); there were no significant differences in IgA and IgM before and after the treatment (P>0.05). After treatment, peripheral blood T-cell subgroup and NK cell in the treatment group increased to a certain extent, which was correlated positively to the therapeutic effect. CONCLUSION: The drug-separated moxibustion at Shenque (CV 8) is a satisfactory method for treatment of ulcerative colitis, and it exerts therapeutic effect possibly through regulating immunological function of the organism.

[Clinical observation on medicine-separated moxibustion for treatment of primary dysmenorrhea and study on the mechanism].

OBJECTIVE: To observe clinical therapeutic effect of medicine-separated moxibustion on primary dysmenorrhea and study on the mechanism. METHODS: Ninety-six cases of primary dysmenorrhea were randomly divided into a treatment group and a control group, 48 cases in each group. They were treated respectively with medicine-separated moxibustion and Yueyueshu Perfusing Powder for 3 consecutive months. RESULTS: The markedly effective and cured rate and the total effective rate were 87.5% and 100.0% in the treatment group, and 29.2% and 83.8% in the control group with significant differences between the two groups (P < 0.01 and P < 0.05). After treatment, blood prosglanding F2 content in menorrhea blood and oxytocin level in plasma during menstruation decreased significantly as compared with those before treatment (P < 0.01). CONCLUSION: The medicine-separated moxibustion has a reliable therapeutic effect on primary dysmenorrhea and the therapeutic effect is exerted possibly by regulating abnormal prosglanding and oxytocin levels in the patient.

[Phenological analysis upon yield and quality of Flos Lonicerae].

Yield and quality of Flos Lonicerae varied in different phenological phase. Yield in the first flowering season was the highest. Yield in the last three flowering season decreased one by one. The dry weight of one thousand flower buds and chlorogenic acid content in the first flowering season were the highest during a year, then was the fourth, the second and the third were both the lower. The difference of above was related to such biologic factors as days of flower bud differentiation, types of flower branch, leaf area, chlorophyll content, and also to such climatic factors as air temperature, hours of sunshine and precipitation.