Genome-wide identification and expression pattern analysis of the Aux/IAA (auxin/indole-3-acetic acid) gene family in alfalfa (Medicago sativa) and the potential functions under drought stress.

BACKGROUND: Auxin/induced-3-acetic acid (Aux/IAA) is an important plant hormone that affects plant growth and resistance to abiotic stresses. Drought stress is a vital factor in reducing plant biomass yield and production quality. Alfalfa (Medicago sativa L.) is the most widely planted leguminous forage and one of the most economically valuable crops in the world. Aux/IAA is one of the early responsive gene families of auxin, playing a crucial role in response to drought stress. However, the characteristics of the Aux/IAA gene family in alfalfa and its potential function in response to drought stress are still unknown. RESULT: A total of 41 Aux/IAA gene members were identified in alfalfa genome. The physicochemical, peptide structure, secondary and tertiary structure analysis of proteins encoded by these genes revealed functional diversity of the MsIAA gene. A phylogenetic analysis classified the MsIAA genes into I-X classes in two subgroups. And according to the gene domain structure, these genes were classified into typical MsIAA and atypical MsIAA. Gene structure analysis showed that the MsIAA genes contained 1-4 related motifs, and except for the third chromosome without MsIAAs, they were all located on 7 chromosomes. The gene duplication analysis revealed that segmental duplication and tandem duplication greatly affected the amplification of the MsIAA genes. Analysis of the Ka/Ks ratio of duplicated MsAux/IAA genes suggested purification selection pressure was high and functional differences were limited. In addition, identification and classification of promoter cis-elements elucidated that MsIAA genes contained numerous elements associated to phytohormone response and abiotic stress response. The prediction protein-protein interaction network showed that there was a complex interaction between the MsAux/IAA genes. Gene expression profiles were tissue-specific, and MsAux/IAA had a broad response to both common abiotic stress (ABA, salt, drought and cold) and heavy metal stress (Al and Pb). Furthermore, the expression patterns analysis of 41 Aux/IAA genes by the quantitative reverse transcription polymerase chain reaction (qRT-PCR) showed that Aux/IAA genes can act as positive or negative factors to regulate the drought resistance in alfalfa. CONCLUSION: This study provides useful information for the alfalfa auxin signaling gene families and candidate evidence for further investigation on the role of Aux/IAA under drought stress. Future studies could further elucidate the functional mechanism of the MsIAA genes response to drought stress.

Genome-wide identification of B-box zinc finger (BBX) gene family in Medicago sativa and their roles in abiotic stress responses.

BACKGROUND: B-box (BBX) family is a class of zinc finger transcription factors (TFs) that play essential roles in regulating plant growth, development, as well as abiotic stress. However, no systematic analysis of BBX genes has yet been conducted in alfalfa (Medica go sativa L.), and their functions have not been elucidated up to now. RESULTS: In this study, 28 MsBBX genes were identified from the alfalfa genome, which were clustered into 4 subfamilies according to an evolutionary tree of BBX proteins. Exon-intron structure and conserved motif analysis reflected the evolutionary conservation of MsBBXs in alfalfa. Collinearity analysis showed that segmental duplication promoted the expansion of the MsBBX family. Analysis of cis-regulatory elements suggested that the MsBBX genes possessed many growth/development-, light-, phytohormone-, and abiotic stress-related elements. MsBBX genes were differentially expressed in leaves, flowers, pre-elongated stems, elongated stems, roots and nodules, and most MsBBXs were remarkably induced by drought, salt and various plant growth regulators (ABA, JA, and SA). Further functional verification demonstrated that overexpressing of the MsBBX11 gene clearly promoted salt tolerance in transgenic Arabidopsis by regulating growth and physiological processes of seedlings. CONCLUSIONS: This research provides insights into further functional research and regulatory mechanisms of MsBBX family genes under abiotic stress of alfalfa.

Metabolic influences on T cell in psoriasis: a literature review.

Psoriasis is a systemic inflammatory disease that frequently coexists with various other conditions, such as essential hypertension, diabetes, metabolic syndrome, and inflammatory bowel disease. The association between these diseases may be attributed to shared inflammatory pathways and abnormal immunomodulatory mechanisms. Furthermore, metabolites also play a regulatory role in the function of different immune cells involved in psoriasis pathogenesis, particularly T lymphocytes. In this review, we have summarized the current research progress on T cell metabolism in psoriasis, encompassing the regulation of metabolites in glucose metabolism, lipid metabolism, amino acid metabolism, and other pathways within T cells affected by psoriasis. We will also explore the interaction and mechanism between psoriatic metabolites and immune cells. Moreover, we further discussed the research progress of metabolomics in psoriasis to gain a deeper understanding of its pathogenesis and identify potential new therapeutic targets through identification of metabolic biomarkers associated with this condition.

Achiral nanoparticle trapping and chiral nanoparticle separating with quasi-BIC metasurface.

Dielectric metasurfaces based on quasi-bound states in the continuum (quasi-BICs) are a promising approach for manipulating light-matter interactions. In this study, we numerically demonstrate the potential of silicon elliptical tetramer dielectric metasurfaces for achirality nanoparticle trapping and chiral nanoparticle separation. We first analyze a symmetric tetramer metasurface, which exhibits dual resonances (P1 and P2) with high electromagnetic field intensity enhancement and a high-quality factor (Q-factor). This metasurface can trap achiral nanoparticles with a maximum optical trapping force of 35 pN for 20 nm particles at an input intensity of 100 mW. We then investigate an asymmetric tetramer metasurface, which can identify and separate enantiomers under the excitation of left-handed circularly polarized (LCP) light. Results show that the chiral optical force can push one enantiomer towards regions of the quasi-BIC system while removing the other. In addition, the proposed asymmetric tetramer metasurface can provide multiple Fano resonances (ranging from R1 to R5) and high trap potential wells of up to 33 kBT. Our results demonstrate that the proposed all-dielectric metasurface has high performance in nanoparticle detection, with potential applications in biology, life science, and applied physics.

Genome-wide identification and expression analysis of the Auxin-Response factor (ARF) gene family in Medicago sativa under abiotic stress.

BACKGROUND: Alfalfa (Medicago sativa) is the most widely planted legume forage and one of the most economically valuable crops in the world. The periodic changes in its growth and development and abiotic stress determine its yield and economic benefits. Auxin controls many aspects of alfalfa growth by regulating gene expression, including organ differentiation and stress response. Auxin response factors (ARF) are transcription factors that play an essential role in auxin signal transduction and regulate the expression of auxin-responsive genes. However, the function of ARF transcription factors is unclear in autotetraploid-cultivated alfalfa. RESULT: A total of 81 ARF were identified in the alfalfa genome in this study. Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were analyzed, identifying that ARF genes are mainly involved in transcriptional regulation and plant hormone signal transduction pathways. Phylogenetic analysis showed that MsARF was divided into four clades: I, II, III, and IV, each containing 52, 13, 7, and 9 genes, respectively. The promoter region of the MsARF gene contained stress-related elements, such as ABRE, TC-rich repeats, MBS, LTR. Proteins encoded by 50 ARF genes were localized in the nucleus without guide peptides, signal peptides, or transmembrane structures, indicating that most MsARF genes are not secreted or transported but only function in the nucleus. Protein structure analysis revealed that the secondary and tertiary structures of the 81 MsARF genes varied. Chromosomal localization analysis showed 81 MsARF genes were unevenly distributed on 25 chromosomes, with the highest distribution on chromosome 5. Furthermore, 14 segmental duplications and two sets of tandem repeats were identified. Expression analysis indicated that the MsARF was differentially expressed in different tissues and under various abiotic stressors. The quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis showed that the expression profiles of 23 MsARF genes were specific to abiotic stresses such as drought, salt, high temperature, and low temperature, as well as tissue-specific and closely related to the duration of stress. CONCLUSION: This study identified MsARF in the cultivated alfalfa genome based on the autotetraploid level, which GO, KEGG analysis, phylogenetic analysis, sequence characteristics, and expression pattern analysis further confirmed. Together, these findings provide clues for further investigation of MsARF functional verification and molecular breeding of alfalfa. This study provides a novel approach to systematically identify and characterize ARF transcription factors in autotetraploid cultivated alfalfa, revealing 23 MsARF genes significantly involved in response to various stresses.

Cross-Breeding Improvement and Performance Analysis of Dominant Production Traits in Grazing-Type Alfalfa (Medicago sativa L.).

Rhizome-rooted Medicago sativa L. "Qingshui" is an excellent germplasm for establishing grazing and ecological grasslands but inferior in yield, in which both high production and ecological values can be achieved by cross-breeding. We have obtained valuable rhizome-rooted hybrid strains (RSA-01, RSA-02, and RSA-03) by crossing of Qingshui and the high-yielding Medicago sativa L. "WL168." In this study, the Qingshui plants with low production performance were crossed for improvement, and progenies with better production and higher quality than those of Qingshui were selected. The results reveal that the branch number, crude protein (CP) content, and relative feed value (RFV) of RSA-01; the stem thickness, CP content, and ether extract (EE) content of RSA-02; and the plant height, stem thickness, branch number, and dry hay yield of RSA-03 were higher than those of Qingshui. Except for the leaf/stem ratio and plant height of RSA-01, leaf/stem ratio of RSA-02, and plant height of RSA-03, the coefficient of variation (CV) of yield traits of the hybrid strains was lower than those of Qingshui, ranging from 0.1% to 4.28%. In addition to the lignin and acid detergent fiber content of RSA-01 as well as EE content of RSA-02 and RSA-03, the CV of the nutritional traits of the hybrid strains was low, ranging from 0.60% to 3.43%. The tested samples were ranked as follows based on yield performance and nutritional values: WL168 > RSA - 03 > RSA - 01 > RSA - 02 > Qingshui and RSA - 01 > WL168 > RSA - 03 > Qingshui > RSA - 02, respectively. Compared with parental Qingshui, RSA-01, RSA-02, and RSA-03 show better yield performance; meanwhile, RSA-01 and RSA-03 had higher nutritional traits. RSA-01 shows heterosis in branch number, CP content, and RFV; RSA-02 shows heterosis in stem thickness and RSA-03 in plant height, stem thickness, branch number, fresh yield, dry hay yield, and CP content. Notably, the low production performance of Qingshui was improved after crossing it with WL168, substantially resulting in an abundant rhizome-rooted germplasm resource for the establishment of grazing grasslands.

Identification and expression analysis of the MADS-box genes of Kentucky bluegrass during inflorescence development.

MADS-box genes play vital roles in multiple biological processes of plants growth and development, especially inflorescence development. In the present study, a comprehensive investigation into the identification and classification of MADS-box genes in Kentucky bluegrass (Poa pratensis) has not been reported. Here, based on the transcriptome of inflorescence, we identified 44 PpMADS-box genes, and gave an overview of the physicochemical properties, phylogeny, protein structures, and potential functions of the proteins encoded by these genes through various bioinformatics software for the first time. Analysis of physicochemical properties revealed that most PpMADS-box were alkaline proteins and possessed similar conserved motifs. Additionally, it was demonstrated that 33 PpMADS-box proteins without signal peptide, leading peptide, transmembrane structure and located in the nucleus were not transported or secreted, so directly played transcriptional regulatory roles in the nucleus. Then, peptide sequences BLAST search and analysis of phylogenetic relationships with MADS-box proteins of P. pratensis, Arabidopsis thaliana, and Oryza sativa were performed. It was found that 44 PpMADS-box proteins were separated into 33 MIKC-type (3 BS, 1 AGL17, 8 AP3/P2, 3 AP1, 5 SEP, 6 SOC and 7 AG genes, respectvely) and 11 type I-type, which include 7 Mγ and 4 Mα. Furthermore, the relative expression levels of the selected 12 genes (MADS3, 15, 16, 17, 18, 20, 24, 27, 30, 36, 38 and 40) at the booting stage, pre-anthesis, anthesis, post-anthesis, and seed filling stage of inflorescences, as well as leaves and roots of the corresponding stages of inflorescences were analyzed, showing that most PpMADS-box genes were highly expressed mainly in young leaves and later inflorescences, and had complex patters in roots. Morever, except for PpMADS30 being highly expressed in the leaves, others were significantly highly expressed in inflorescence and/ or roots, demonstrating PpMADS-box genes also regulate leaves and roots development in plant. This study provides valuable insights into the MADS-box family genes in Kentucky bluegrass and its potential functional characteristics, expression pattern, and evolution in floral organogenesis and even reproduction development. @media print { .ms-editor-squiggler { display:none !important; } } .ms-editor-squiggler { all: initial; display: block !important; height: 0px !important; width: 0px !important; }. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01216-1.

Hsa_circ_0097922 promotes tamoxifen resistance and cell malignant behaviour of breast cancer cells by regulating ACTN4 expression via miR-876-3p.

An increasing number of findings have verified the critical roles of circular RNAs (circRNAs) in human cancers, and chemotherapy resistance is a poor prognostic factor for breast cancer (BC). This study is designed to explore the function of hsa_circ_0097922 in the tamoxifen resistance of breast cancer. Hsa_circ_0097922, microRNA-876-3p (miR-876-3p), and alpha-actinin 4 (ACTN4) level were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Cell survival, proliferation, apoptosis, migration and invasion were detected by Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), and flow cytometry, wound healing and Transwell assays. Protein levels of proliferating cell nuclear antigen (PCNA), B-cell lymphoma-2 (Bcl-2), cleaved caspase 3, matrix metalloproteinase 9 (MMP9), and ACTN4 were determined using western blot assay. Using bioinformatics software, the binding between miR-876-3p and hsa_circ_0097922 or ACTN4 was predicted, followed by confirmation by RNA immunoprecipitation (RIP) and RNA pull-down assays. A xenograft tumour model in vivo analysed the biological role of hsa_circ_0097922 on BC tumour growth and drug resistance. Hsa_circ_0097922 and ACTN4 were increased, and miR-876-3p was decreased in tamoxifen resistance BC cells. Moreover, hsa_circ_0097922 knockdown can block BC cell malignant behaviour and tamoxifen resistance in vitro. Mechanically, hsa_circ_0097922 acted as a sponge of miR-876-3p to regulate ACTN4 expression. Hsa_circ_0097922 silencing increased the drug sensitivity of BC in vivo. Hsa_circ_0097922 might regulate BC cell malignant behaviour and tamoxifen resistance partly by regulating the miR-876-3p/ACTN4 axis, hinting at a promising therapeutic target for the BC treatment.

Rigid ureteroscopic lithotripsy in the lateral decubitus position for upper urinary tract stones.

BACKGROUND: The current study aimed to assess a novel ureteroscopic technique developed for treating upper urinary calculi based on a specially designed lateral decubitus body position that could avoid stone loss by adjusting to the effects of gravity. METHODS: This retrospective study examined patients with upper urinary calculi who were surgically treated from November 2008 to January 2020, using a new body position and a rigid ureteroscope. Clinical outcomes, stone-free rates, operative times and complications were evaluated, and factors that could influence treatment success were determined. RESULTS: In total, 1080 patients were included, and 1145 operations were performed. The maximum calculus diameters were 11.22 ± 5.01 mm. Operative times were 48.60 ± 27.44 min. A total of 1042 cases were successfully treated, with a stone-free rate of 91.00%. Multivariate analysis showed that female sex (OR = 2.135, 95% CI 1.332-3.422, P = 0.002), thin scope standby (OR = 1.643, 95% CI 1.074-2.514, P = 0.022), laser lithotripsy (OR = 5.087, 95% CI 2.400-10.785, P = 0.000) and stone size (OR = 0.946, 95% CI 0.912-0.981, P = 0.003) were independently associated with stone-free outcomes. In total, 2 ureteral perforations, 2 ureteric avulsions and 4 urosepsis cases were observed, but were all cured without sequelae. CONCLUSIONS: Ureteroscopic lithotripsy in the lateral decubitus position is a safe and effective technique for treating upper urinary tract calculi, especially upper ureteral calculi.

Analysis on characteristics of female gametophyte and functional identification of genes related to inflorescences development of Kentucky bluegrass.

The inflorescence is composed of spikes, and the spike is the carrier of grass seed formation and development, so the development status of inflorescence implies grass seed yield and quality. So far, the systematic analysis of inflorescence development of Kentucky bluegrass has not been reported. The development process of the female gametophyte of wild germplasm materials of Kentucky bluegrass in Gannan, Gansu Province of China (KB-GN), was observed. Based on this, the key developmental stages of inflorescence in KB-GN were divided into premeiosis (GPreM), meiosis (GM), postmeiosis (GPostM), and anthesis (GA), and four stages of inflorescence were selected to analyze the transcriptome expression profile. We found that its sexual reproduction formed a polygonum-type embryo sac. Transcriptome analysis showed that 4256, 1125, 1699, and 3127 genes were highly expressed in GPreM, GM, GPostM, and GA, respectively. And a large number of transcription factors (TFs) such as MADS-box, MYB and NAC, AP2, C2H2, FAR1, B3, bHLH, WRKY, and TCP were highly expressed throughout the inflorescence development stages. KEGG enrichment and MapMan analysis showed that genes involved in plant hormone metabolism were also highly expressed at the entire stages of inflorescence development. However, a few TFs belong to stage-specific genes, such as TRAF proteins with unknown function in plants was screened firstly, which was specifically and highly expressed in the GPreM, indicating that TRAF may regulate the preparatory events of meiosis or be essential for the development of megaspore mother cell (MMC). The expression patterns of 15 MADS-box genes were analyzed by qRT-PCR, and the expression results were consistent with that of the transcriptome. The study on the inflorescence development of KB-GN will be great significant works and contribution to illustrate the basic mechanism of grass seeds formation and development.

Molecular genotyping of Candida albicans isolated from different sites may trace the etiological infection routes: Observation in China.

BACKGROUND: Invasive candidiasis is a growing concern worldwide, especially in immunocompromised patients, including ICU patients. OBJECTIVES: As Candida albicans is the leading cause of candidaemia, it is important to investigate the evolution of C. albicans in patients with candidaemia. METHODS: We analysed 238 strains of C. albicans isolated from different body sites. Antifungal susceptibility testing, CAI loci genotyping and multilocus sequence typing (MLST) of all isolates were performed. The relationships among the total isolates that differed in sequence at only one of the seven housekeeping gene loci were analysed using eBURST. RESULTS: Multilocus sequence typing analysis in 238 isolates by combining seven housekeeping alleles revealed 175 diploid sequence types, in which 84 were newly identified. eBURST analysis for these data recognised 19 clonal complexes (CCs) and 79 singletons. Besides, seventy-three CAI genotypes were identified. Blood isolates showed maximum genotypes (49), and the dominant genotypes were CAI 17-21 and CAI 21-21. Oral isolates possessed 25 CAI genotypes, and the dominant genotypes were CAI 17-21 and CAI 21-21 as well. Since isolates with CAI allele numbers <30 showed easier transmission, CAI 17-21 and CAI 21-21 were the most frequently transmitted. Finally, the CAI genotypes were classified into six groups. CONCLUSIONS: This work revealed the oral and blood strains isolated from the patients with candidaemia in ICU shared the identical dominant CAI genotypes. Our data expanded the C. albicans MLST database and helped with understanding the evolution and spread of invasive candidiasis.

Fixed Cutaneous Sporotrichosis Due to Sporothrix globosa.

PURPOSE: This report describes a case of a skin sporotrichosis infection and the steps taken to identify an effective antifungal treatment. PATIENTS AND METHODS: A 50-year-old woman from Jilin province, China, presented complaining of a small mass that had been on her right upper eyelid for two years. A skin biopsy was taken and submitted for bacterial and mycological assessment. Bacterial culture from the lesion was negative, but a fungal culture was positive. In vitro susceptibility test was performed to assess its susceptibility to antifungal drugs. RESULTS: The skin biopsy showed infectious granuloma. Fungal culture was identified as Sporothrix globosa based on both the morphological features and confirmation by the molecular method; it was resistant to many kinds of antifungal drugs, including amphotericin B, voriconazole, fluconazole, and caspofungin. However, it was relatively sensitive to itraconazole. The patient was prescribed 0.2 g itraconazole to be taken twice per day. One month later, she had almost completely recovered from her symptoms. The treatment lasted for 3 months and her liver function and renal function were normal at the endpoint. CONCLUSION: Itraconazole was an effective treatment in this case of a multidrug-resistant sporotrichosis caused by S. globosa.

Comprehensive assessment of the association between XPC rs2228000 and cancer susceptibility based on 26835 cancer cases and 37069 controls.

Objectives In the present study, we examined available articles from online databases to comprehensively investigate the effect of the XPC (xeroderma pigmentosum complementation group C) rs2228000 polymorphism on the risk of different types of clinical cancer. Methods We conducted a group of overall and subgroup pooling analyses after retrieving the data from four databases (updated till September 2019). The P-value of association, OR (odds ratios), and 95% CI (confidence interval) were calculated. Results We selected a total of 71 eligible studies with 26835 cancer cases and 37069 controls from the 1186 retrieved articles. There is an enhanced susceptibility for bladder cancer cases under T vs. C [P=0.004; OR (95% CI) = 1.25 (1.07, 1.45)], TT vs. CC [P=0.001; 1.68 (1.25, 2.26)], CT+TT vs. CC [P=0.016; 1.26 (1.04, 1.53)], and TT vs. CC+ CT [P=0.001; 1.49 (1.18, 1.90)] compared with negative controls. Additionally, there is an increased risk of breast cancer under T vs. C, TT vs. CC and TT vs. CC+ CT (P<0.05, OR > 1). Nevertheless, there is a decreased risk of gastric cancer cases in China under T vs. C [P=0.020; 0.92 (0.85, 0.99)], CT vs. CC [P=0.001, 0.83 (0.73, 0.93)], and CT+TT vs. CC [P=0.003, 0.84 (0.76, 0.94)]. Conclusions The TT genotype of XPC rs2228000 may be linked to an increased risk of bladder and breast cancer, whereas the CT genotype is likely to be associated with reduced susceptibility to gastric cancer in the Chinese population.

Elementary screening of lymph node metastatic-related genes in gastric cancer based on the co-expression network of messenger RNA, microRNA and long non-coding RNA.

Gastric cancer (GC) is the fifth most common cancer and the third leading cause of cancer-related deaths worldwide. The high mortality might be attributed to delay in detection and is closely related to lymph node metastasis. Therefore, it is of great importance to explore the mechanism of lymph node metastasis and find strategies to block GC metastasis. Messenger RNA (mRNA), microRNA (miRNA) and long non-coding RNA (lncRNA) expression data and clinical data were downloaded from The Cancer Genome Atlas (TCGA) database. A total of 908 differentially expressed factors with variance >0.5 including 542 genes, 42 miRNA, and 324 lncRNA were screened using significant analysis microarray algorithm, and interaction networks were constructed using these differentially expressed factors. Furthermore, we conducted functional modules analysis in the network, and found that yellow and turquoise modules could separate samples efficiently. The groups classified in the yellow and turquoise modules had a significant difference in survival time, which was verified in another independent GC mRNA dataset (GSE62254). The results suggested that differentially expressed factors in the yellow and turquoise modules may participate in lymph node metastasis of GC and could be applied as potential biomarkers or therapeutic targets for GC.

Elementary screening of lymph node metastatic-related genes in gastric cancer based on the co-expression network of messenger RNA, microRNA and long non-coding RNA

Gastric cancer (GC) is the fifth most common cancer and the third leading cause of cancer-related deaths worldwide. The high mortality might be attributed to delay in detection and is closely related to lymph node metastasis. Therefore, it is of great importance to explore the mechanism of lymph node metastasis and find strategies to block GC metastasis. Messenger RNA (mRNA), microRNA (miRNA) and long non-coding RNA (lncRNA) expression data and clinical data were downloaded from The Cancer Genome Atlas (TCGA) database. A total of 908 differentially expressed factors with variance >0.5 including 542 genes, 42 miRNA, and 324 lncRNA were screened using significant analysis microarray algorithm, and interaction networks were constructed using these differentially expressed factors. Furthermore, we conducted functional modules analysis in the network, and found that yellow and turquoise modules could separate samples efficiently. The groups classified in the yellow and turquoise modules had a significant difference in survival time, which was verified in another independent GC mRNA dataset (GSE62254). The results suggested that differentially expressed factors in the yellow and turquoise modules may participate in lymph node metastasis of GC and could be applied as potential biomarkers or therapeutic targets for GC.

The A395T mutation in ERG11 gene confers fluconazole resistance in Candida tropicalis causing candidemia.

The mechanism of fluconazole resistance in Candida tropicalis is still unclear. Recently, we isolated a fluconazole-resistant strain of C. tropicalis from the blood specimen of a patient with candidemia in China. In vitro antifungal susceptibility of the isolate was determined by using CLSI M27-A3 and E-test methods. The sequence of ERG11 gene was then analyzed, and the three-dimensional model of Erg11p encoded by ERG11 gene was also investigated. The sequencing of ERG11 gene revealed the mutation of A395T in this fluconazole-resistant isolate of C. tropicalis, resulting in the Y132F substitution in Erg11p. Sequence alignment and three-dimensional model comparison of Erg11ps showed high similarity between fluconazole-susceptible isolates of C. tropicalis and Candida albicans. The comparison of the three-dimensional models of Erg11ps demonstrated that the position of the Y132F substitution in this isolate of C. tropicalis is identical to the isolate of C. albicans with fluconazole resistance resulting from Y132F substitution in Erg11p. Hence, we ascertain that the Y132F substitution of Erg11p caused by A395T mutation in ERG11 gene confers the fluconazole resistance in C. tropicalis.

New potential solutions for the chemolysis of urinary phosphate calculi determined by an in vitro study.

To find a more efficient solution for chemolysis of urinary calculi, several organic acids were chosen to form solutions by consulting the composition of a classic solution, Suby G. The solutions together with Renacidin, another classic solution, were designed to react with the 4 phosphate components of urinary stone. The processes were real-time measured and analysed by a focused beam reflectance measurement, and the efficiency factors were investigated and discussed in detail. The results show that several organic acids, e.g. hydroxyacetic acid, lactic acid and α-ketoglutaric acid, are more efficient than citric acid in dissolving urinary phosphate calculus. The new solutions containing the organic acids are promising for improving chemolysis treatment.

Antifungal activity of geldanamycin alone or in combination with fluconazole against Candida species.

A standardized broth microdilution method was used to test the antifungal activity of geldanamycin (GA), an inhibitor of heat shock protein 90 (Hsp90), alone or in combination with the antifungal agent fluconazole (FLC) against 32 clinical isolates of Candida spp. In addition, a disk diffusion test was also used to evaluate the antifungal effect of these two drugs against Candida spp. by measuring the inhibition zone diameters. We found that the range of minimal inhibitory concentrations (MICs) for GA alone against Candida spp. was 3.2-12.8 mg/L and the geometric mean of MICs was 6.54 mg/L. In addition, the combination of GA with FLC showed synergistic effects in vitro against 2 FLC-susceptible and 6 FLC-resistant isolates of C. albicans. As for the other isolates, indifference but no antagonism was observed. In the disk diffusion assay, the diameter of inhibition zones for FLC combined with GA against FLC-resistant C. albicans isolates was 30 mm, while no inhibition was observed with FLC alone. These results demonstrate that GA possesses antifungal activity against Candida spp., and the combination of GA with FLC shows in vitro synergistic activity against some C. albicans isolates, especially those resistant to FLC.

Epidemiologic and economic burden of influenza in the outpatient setting: a prospective study in a subtropical area of China.

OBJECTIVES: To understand the incidence of outpatient influenza cases in a subtropical area of China and the associated economic burden on patients' families. METHODS: A hospital-based prospective study was conducted in Zhuhai City during 2008-2009. All outpatient influenza-like illness (ILI) cases were identified in 28 sentinel hospitals. A representative sample of throat swabs from ILI cases were collected for virus isolation using Madin-Darby canine kidney cells. The incidence of outpatient influenza cases in Zhuhai was estimated on the basis of the number of influenza patients detected by the sentinel sites. A telephone survey on the direct costs associated with illness was conducted as a follow-up. RESULTS: The incidence of influenza was estimated to be 4.1 per 1,000 population in 2008 and 19.2 per 1,000 population in 2009. Children aged <5 years were the most-affected population, suffering from influenza at the highest rates (34.3 per 1,000 population in 2008 and 95.3 per 1,000 population in 2009). A high incidence of 29.2-40.9 per 1000 population was also seen in young people aged 5-24 years in 2009. ILI activity and influenza virus isolations adopted a consistent seasonal pattern, with a summer peak in July 2008 and the longest epidemic period lasting from July-December 2009. The medical costs per episode of influenza among urban patients were higher than those for rural patients. A total of $1.1 million in direct economic losses were estimated to be associated with outpatient influenza during 2008-2009 in Zhuhai community. CONCLUSIONS: Influenza attacks children aged <5 years in greater proportions than children in other age groups. Seasonal influenza 2008 and Pandemic influenza A (H1N1) 2009 had different epidemiological and etiological characteristics. Direct costs (mostly medical costs) impose an enormous burden on the patient family. Vaccination strategies for high-risk groups need to be further strengthened.