Integrative transcriptome, proteome, phosphoproteome and genetic mapping reveals new aspects in a fiberless mutant of cotton.

To investigate the molecular mechanisms of fiber initiation in cotton (Gossypium spp.), an integrated approach combining transcriptome, iTRAQ-based proteome and genetic mapping was taken to compare the ovules of the Xuzhou 142 wild type (WT) with its fuzzless-lintless (fl) mutant at -3 and 0 day post-anthesis. A total of 1,953 mRNAs, 187 proteins, and 131 phosphoproteins were differentially expressed (DE) between WT and fl, and the levels of transcripts and their encoded proteins and phosphoproteins were highly congruent. A functional analysis suggested that the abundance of proteins were mainly involved in amino sugar, nucleotide sugar and fatty acid metabolism, one carbon pool for folate metabolism and flavonoid biosynthesis. qRT-PCR, Western blotting, and enzymatic assays were performed to confirm the regulation of these transcripts and proteins. A molecular mapping located the lintless gene li3 in the fl mutant on chromosome 26 for the first time. A further in-silico physical mapping of DE genes with sequence variations between fl and WT identified one and four candidate genes in the li3 and n2 regions, respectively. Taken together, the transcript abundance, phosphorylation status of proteins at the fiber initiation stage and candidate genes have provided insights into regulatory processes underlying cotton fiber initiation.

[Genome-wide analysis of the LPAAT gene family in Gossypium raimondii and G. arboreum, and expression analysis of its orthologs in G. hirsutum].

Lysophosphatidic acid acyltransferase (LPAAT) which converts lysophosphatidic acid into phosphatidic acid is a key enzyme in biosynthesis pathway of lipid in plants. In this study, we identified 17 members of the LPAAT gene family from genomic data of G. raimondii-D5 and G. arboreum-A2. Analysis of gene structure, chromosome distribution and phylogenetic evolution of LPAAT genes in diploid Gossypium using bioinformatics approaches showed that these genes can be divided into distinct subfamilies based on the distance of their genetic relationship. Moreover, the gene structures were similar within LPAAT subfamily members. The amino acid sequences encoded by LPAAT family genes contained three conserved motifs, including ΦFPEGTR-G binding site and Φ-NHQS- ΦDΦΦ catalytic site. Phylogenetic analysis of LPAAT gene family demonstrated significant differences in evolution of LPAAT in different species. Finally, expression analysis of G. hirsutum ovules in different stages from RNA-seq and qRT-PCR data indicated that LPAAT gene may play a positive role in oil accumulation. Our studies facilitate understanding of the function of LPAAT gene family in Gossypium and selecting better LPAAT genes for further functional validation.

In vitro evaluation of antiviral activity of tea seed saponins against porcine reproductive and respiratory syndrome virus.

BACKGROUND: Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the major swine pathogens. This virus causes immune suppression and other secondary infections, leading to significant economic losses in the swine industry. Tea seed saponins (TS) are a natural extract from tea seeds with anti-cancer, anti-inflammatory and antiviral activity. In this study, we demonstrated that TS possessed anti-PRRSV activity. METHODS: MTT assay and trypan blue staining were used to evaluate the cytotoxicity and antiviral ability of TS in cell culture. Apoptosis was measured to assess the safety of TS on Marc-145 cells. Time-of-addition assay, entry inhibition assay and virucidal assay were used to assess the antiviral action of TS. The effect of TS on host cellular gene expression was analysed by real-time PCR. Absolute quantification RT-PCR and western blot were used to study the inhibitory effect of TS on PRRSV N gene and protein expression. RESULTS: Our results showed that 50% cytotoxic concentrations (CC50) and 50% effective concentration (EC50) of TS were 59.86 ±0.3841 µg/ml and 24.29 ±1.194 µg/ml, respectively. The maximum non-cytotoxic concentration of TS on Marc-145 cells was 30 µg/ml. TS inhibited PRRSV-induced cell apoptosis and effectively inhibited PRRSV replication by reducing the expression of host cellular gene PABP, and significantly inhibited virus N gene/protein expression. CONCLUSIONS: TS possessed anti-PRRSV activity in vitro and could serve as a potential antiviral drug for PRRSV prevention and control.

Severe burn injury in late pregnancy: a case report and literature review.

The management of serious burn injuries during pregnancy is an unsolved clinical problem because of the low incidence of this disease. Although it has been documented that the effect of burns on fetal and maternal survival is detrimental, there have been conflicting reports among the different burn centers regarding the mortality of burned pregnant women and the management of burn patients during pregnancy. We report a case of severe burn in late pregnancy treated at our burn center. Additionally, we searched and summarized the literature concerning the management of pregnant patients to provide useful information for their treatment.

Sodium tanshinone IIA sulfonate inhibits the meq, ul49 and VP22 expression of Marek's disease virus.

BACKGROUND: Our previous studies have demonstrated that sodium tanshinone IIA sulfonate (STS), a natural compound derived from Salviae Miltiorrhizae Radix et Rhizoma (Danshen), could effectively inhibit Marek's disease virus (MDV) infection both in vitro and in vivo, but the underlying mechanisms remain unclear. The main objective of the study was to explore the effect of STS on the meq, ul49 and VP22 expression of MDV in vitro. METHODS: Quantitative real-time PCR (qRT-PCR) was used to analyse the effect of STS on meq and ul49 expression at both the DNA and messenger RNA (mRNA) level, and the effect of STS on VP22 was assessed by immunofluorescence assay and western blotting. RESULTS: The DNA and mRNA copy numbers of meq and ul49 significantly decreased in the groups treated with STS compared with MDV control (P<0.05), which indicated that STS could inhibit the expression of meq and ul49 at both the DNA and mRNA level. Moreover, the expression of VP22 encoded by ul49 was also significantly inhibited (P<0.05). CONCLUSIONS: STS possessed anti-MDV activity in chicken embryo fibroblasts. Its antiviral mechanisms may be ascribed to inactivating MDV directly, disturbing meq and ul49 replication and inhibiting the expression of VP22 encoded by ul49. These results suggested that STS is a promising natural compound to be further developed as an antiviral agent against MDV infection.

Epidemiological and genetic diversity of Staphylococcus aureus causing bloodstream infection in Shanghai, 2009-2011.

OBJECTIVES: Staphylococcus aureus or methicillin-resistant Staphylococcus aureus (MRSA) has been an important pathogen causing bloodstream infections. Our study aimed to investigate the epidemiological and genetic diversity of clinical S. aureus isolates from patients with bloodstream infection in four hospitals of Shanghai from 2009 to 2011. METHODS: A collection of S. aureus isolates causing bloodstream infection from four hospitals in the central part of Shanghai was carried out. Antimicrobial susceptibility testings of collected isolates were performed according to the Clinical and Laboratory Standards Institute (CLSI) guidelines, and spa-type, multi-locus sequence typing, agr type and toxin gene profiling were performed to explore the molecular diversity. Moreover, MRSA strains were also characterized by Staphylococcal cassette chromosome mec (SCCmec) typing. RESULTS: The drugs such as linezolid, teicoplanin and vancomycin were efficacious for treating S. aureus including MRSA bloodstream infection. Methicillin-sensitive Staphylococcus aureus (MSSA) strains displayed distinct diversity in molecular characterization and toxin genes, and three virulent MSSA strains encoding at least five toxins were detected. Five community-associated MRSA (CA-MRSA) strains were found, but the majority (88.7%) of MRSA strains belonged to two epidemic clones (ST239-MRSA- III and ST5-MRSA- II) with different toxin gene profiles among patients with bloodstream infection. CONCLUSIONS: Healthcare-associated MRSA (HA-MRSA) strains were still the main pathogen causing bloodstream infections in spite of the emergence of CA-MRSA strains in hospital setting.

[Multi-center clinical trial of FLAMIGEL (hydrogel dressing) for the treatment of residual burn wound].

OBJECTIVE: To evaluate the effect of FLAMIGEL (hydrogel dressing) on the repair of residual burn wound. METHODS: Sixty burn patients with residual wounds hospitalized in 6 burn units from November 2011 to May 2012 were enrolled in the multi-center, randomized, and self-control clinical trial. Two residual wounds of each patient were divided into groups T (treated with FLAMIGEL) and C (treated with iodophor gauze) according to the random number table. On post treatment day (PTD) 7 and 14, wound healing rate was calculated, with the number of completely healed wound counted. The degree of pain patient felt during dressing change was evaluated using the visual analogue scale (VAS). The mean numbers of wounds with score equal to zero, more than zero and less than or equal to 3, more than 3 and less than or equal to 6, more than 6 and less than or equal to 10 were recorded respectively. Wound secretion or exudate samples were collected for bacterial culture, and the side effect was observed. Data were processed with repeated measure analysis of variance, t test, chi-square test, and nonparametric rank sum test. RESULTS: Wound healing rate of groups T, C on PTD 7 was respectively (67 ± 24)%, (45 ± 25)%, and it was respectively (92 ± 16)%, (72 ± 23)% on PTD 14. There was statistically significant difference in wound healing rate on PTD 7, 14 between group T and group C (F = 32.388, P < 0.01). Ten wounds in group T and four wounds in group C were healed completely on PTD 7, with no significant difference between them (χ(2) = 0, P > 0.05). Forty-two wounds in group T and seven wounds in group C healed completely on PTD 14, with statistically significant difference between them (χ(2) = 42.254, P < 0.01). Patients in group T felt mild pain during dressing change for 37 wounds, with VAS score higher than zero and lower than or equal to 3. Evident pain was observed in patients of group C during dressing change for 43 wounds, and it scored higher than 3 and less than or equal to 6 by VAS evaluation. There was statistically significant difference in mean number of wounds with different grade of VAS score between group T and group C (Z = -4.638, P < 0.01). Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, E. coli, Baumanii, and Staphylococcus epidermidis were all detected in both groups, but there was no statistical difference between group T and group C (χ(2) = 0.051, P > 0.05). No side effect was observed in either of the two groups during the whole trial. CONCLUSIONS: FLAMIGEL can accelerate the healing of residual burn wounds and obviously relieve painful sensation during dressing change.

Molecular epidemiologic analysis of Staphylococcus aureus isolated from four burn centers.

Staphylococcus aureus has been a major cause of hospital-acquired infections. Methicillin-resistant Staphylococcus aureus (MRSA) has emerged since 1980s as an epidemiologic problem in hospitals. This old pathogen brings a new challenge to all physicians and bacteriologists. Hence, effective measures of MRSA control are in critical need. S. aureus or MRSA is one of the leading causes of infection among burn centers, resulting in a number of poor outcomes and even death. The present study performed a molecular epidemiologic analysis of S. aureus isolated from four burn centers in the southeast of China. A total of 85 isolates were collected, and molecular characters were determined for further investigation. In this study, the prevalent clone of MRSA among four burn centers was found to be SCCmec III (spa-type t030, agr I), which is resistant to 4 kinds of antimicrobials including erythromycin, clindamycin, kanamycin and mupirocin. Discrepancy between mecA detection and conventional tests used for MRSA identification was observed unintentionally. Our data demonstrated that the overall prevalence rate of MRSA was 55.3%, and drugs such as sulfamethoxazole/trimethoprim, linezolid and fusidic acid are efficient antibiotic options for treating S. aureus or MRSA infections among four burn centers studied in present investigation.

[Analysis of the molecular epidemiology and distribution of pathogenic bacteria in burn wards of Ruijin Hospital from 2004 to 2006].

OBJECTIVE: To analyse the distribution, drug resistance and epidemiology of pathogenic bacteria in the burn wards of Ruijin Hospital. METHODS: Seventeen strains of Methicillin resistant staphylococcus aureus (MRSA), 52 strains of Pseudomonas aeruginosa (PA), and 11 strains of Acinetobacter baumannii (AB) isolated from the wound secretion, venous catheters, blood, urine and stool etc. were collected from burn patients hospitalized in our department from January 2004 to December 2006. The distribution and the drug resistance profile of bacteria were analyzed, and the homology analysis was performed by randomly amplified polymorphic DNA (RAPD). RESULTS: MRSA, PA and AB were the major strains in our burn wards in recent years, of which Staphylococcus aureus (SA) was the most dominant. During these 3 years, MRSA accounted for 77% (63/82), 85% (63/74), and 75% (74/99), respectively, for SA isolated in this period. MRSA was resistant to Amikacin, Gentamicin, Erythromycin, Clindamycin and Levofloxacin; PA was resistant to Amikacin, Gentamicin, Piperacillin, Ceftazidime, Cefoperazone, Aztreonam and Imipenem; AB was resistant to Amikacin, Gentamicin, Piperacillin, Ceftazidime, Imipenem and Ciprofloxacin. Three bacteria were found to belong to the same type in the RAPD homology analysis. CONCLUSIONS: There are many kind of multi-drug resistant pathogenic bacteria for nosocomial infection in our burn wards. To control the spread of infection due to above-mentioned 3 bacteria is the focus of nosocomial infection control.

[Protective effect of lytic cocktail (inhibition of stress) on lung injury in severe burn rat].

OBJECTIVE: To investigate the protective effect of inhibition of stress (lytic cocktail) on lung injury in severe burn rats at early stage. METHODS: Sprague-Dawley rats inflicted with 30% TBSA full-thickness burn were randomly divided into A group (n = 36, fluid resuscitation with administration of lytic cocktail), B group (n = 36, fluid resuscitation only). Lung function was evaluated by partial pressure of oxygen (PaO2) in arterial blood and histopathologic changes on 3, 5, 7, 10 post burn day (PBD). The levels of malonyldialdehyde (MDA), myeloperoxidase (MPO), tumor necrosis factor-alpha(TNF-alpha) and interferon-gamma (IFN-gamma) in lung tissue were measured at the same time points. RESULTS: The PaO2 level in A group on 3 PBD (12.58 +/- 0.41 kPa) was significantly higher than that in B group (8.86 +/- 0.23 kPa, P < 0.01). Compared with those in B group, the levels of MDA and MPO were significantly decreased in A group at each time point (P < 0.05 or 0.01), the levels of TNF-alpha on 3, 5, 7 PBD (P < 0.05 or 0.01) and IFN-gamma on 5, 7, 14 PBD (P < 0.01) were also decreased in A group. Swollen lung mesenchyme was alleviated, infiltration of inflammatory cell was lessened in A group. CONCLUSION: Lytic cocktail combined with immediate fluid resuscitation can inhibit stess response, downregulate the expression of inflammatory factor, ameliorate lung function in severe burn rat at early stage.

[Analysis of methicillin resistant Staphylococcus aureus in burn ward of Ruijin hospital].

OBJECTIVE: To analyze the epidemiology of methicillin resistant Staphylococcus aureus (MRSA) in molecular level in burn centre of Shanghai Ruijin hospital. METHODS: The vicissitude of Staphylococcus aureus in the burn centre from 2003 to 2005 was analyzed with software WHONET5. Multiprimer random amplified polymorphic DNA(RAPD) was used to analyze the homology of 17 MRSA strains. RESULTS: RAPD analysis (primer ERIC2 and RAPD7) showed that all 17 MRSA strains were identical (Burn-A type). CONCLUSION: MRSA with same RAPD type is prevalent in our burn centre for many years, so emphasis should be laid on the anti-infection therapy and its cross infection control. Staphylococcus aureus;

[Influence of stress inhibition on invasive infection in burn wound during early postburn stage in severely burned rats].

OBJECTIVE: To investigate the influence of inhibition of stress on the survival rate, organ dysfunction and (Th)1/Th2 cytokine profiles of the rats with invasive infection in the wound at early postburn stage. METHODS: Sprague-Dawley rats inflicted with 30% TBSA full thickness burn were randomly divided into A (n = 36, with immediate resuscitation), B (n = 36, with immediate resuscitation and lytic cocktail administration). After subeschar injection of 0.1 ml Pseudomonas aeruginosa (10(8) CFU/ml) on 3rd postburn day, the subeschar bacterial quantitative analysis, the survival rate at 96 hours after bacteria injection, the parameters of organ dysfunction and the mRNA expression of IL-2, IL-4, IL-10 and IFN-gamma were determined by corresponding methods. RESULTS: The quantity of subeschar bacteria was larger than 1 x 10(5)/gram in both groups. The survival rate in B group (66.7 +/- 2.6)% was obviously higher than that in A group (33.3 +/- 1.7)%, (P < 0.01). Inflammatory infiltration and pathological changes in the internal organs in B group were alleviated obviously compared with A group. The expression of IL-2 mRNA in B group was significantly lower than that in A group before bacterial inoculation, but increased at 48 and 96 hours after bacterial inoculation, while it was lowered in A group at the same time points (P < 0.05). The expression of IFN-gamma mRNA in A group was significantly lower than that in B group (P < 0.01), while that of IL-4 and IL-10 mRNA in A group was evidently higher than that in B group (P < 0.05 approximately 0.01). CONCLUSION: Inhibition of the stress response during early postburn stage could be beneficial to the prevention of the bacterial invasion due to the changes in Th1/Th2 ratio.