Microbiology rapid on-site evaluation: a better method for Mucoid Pseudomonas Aeruginosa diagnosis in bronchiectasic patients.

OBJECTIVE: The aim of this study is to find an accurate and fast method to diagnose the pathogen of bronchiectasis. PATIENTS AND METHODS: Ten bronchiectasic patients diagnosed with Mucoid Pseudomonas Aeruginosa (MPA) in the past two years were analyzed. Accuracy and time were compared between microbiology rapid on-site evaluation (M-ROSE) and sputum bacterial culture. RESULTS: The accuracy rate of M-ROSE in the patients is 100% consistent with bacterial culture results. The average time of M-ROSE is about 4.3 min, which is over 1000 times shorter than that of sputum bacterial culture. CONCLUSIONS: M-ROSE may be a better method for etiological diagnosis of MPA.

Tobacco control in China and the road to Healthy China 2030.

Healthy China 2030 aims to reduce the adult smoking rate from 27.7% in 2015 to 20% by 2030. Achieving this goal requires a review of the tobacco control measures introduced in China to date, the gaps that remain and the opportunities ahead. In 2008, the World Health Organization introduced six measures to reduce demand for tobacco called MPOWER. The progress China has made in implementing these measure varies: 1) monitor tobacco use and prevention policies. The surveillance on tobacco use has been rigorous, but the monitoring and evaluation of tobacco control policies needs to be strengthened; 2) protect people from tobacco use: pushes for national tobacco control legislation have stalled, but 18 subnational legislations have passed; 3) offer help to quit tobacco use. The accessibility and quality of cessation services needs to be improved; 4) warn about the dangers of tobacco. While there are no pictorial health warnings, tobacco control advocates have launched a series of anti-smoking media campaigns to inform the public; 5) enforce bans on tobacco advertising, promotion, and sponsorship. Legal loopholes and poor enforcement remain challenges; 6) raise taxes on tobacco: cigarettes in China are relatively cheap and increasingly affordable, which demonstrates the need for further tobacco tax increases indexed to inflation and income. China maintains a tobacco monopoly that interferes with tobacco control efforts and fails to regulate tobacco products from the public health perspective. Effective MPOWER measures, which depend upon the removal of tobacco industry interference from policymaking, are key to achieving the goal set by Healthy China 2030.

Rectal Temperature of Corpse and Estimation of Postmortem Interval.

ABSTRACT: Measurement of corpse temperature is mainly used for estimation of early postmortem interval, and rectal temperature is often used as a representative of body's core temperature in actual work because it is simple, quick and non-invasive. At present, the rectal temperature postmortem interval estimation method internationally accepted and widely used is HENSSGE's nomogram method, while many domestic scholars also deduced their own regression equations through a large number of case data. Estimation of postmortem interval based on rectal temperature still needs further study. The nomogram method needs to be optimized and extended, and quantification of its influencing factors needs to be dealt with more scientifically. There is still a lack of consensus on the probability and duration of the temperature plateau. There is no clear understanding of the probability and extent of the change in initial temperature caused by various causes. New methods and ideas enrich methodological research, but it still lacks systemicity and practicality. This article reviews the researches on estimation of postmortem interval based on rectal temperature in order to summarize the current situation of previous researches and seek new breakthrough points. Because the decline of body temperature can be easily influenced by many factors in vitro and vivo, and the influencing factors in different regions vary greatly, regionalization research and application may be a practical exploration to improve the accuracy of postmortem interval determination.

Enhanced hydrogen adsorption on graphene by manganese and manganese vanadium alloy decoration.

In this work, two kinds of novel manganese decorated (G + Mn) and manganese-vanadium co-decorated (G + MnV) graphene composites are synthesized by in situ wet chemical reduction, and their hydrogen storage properties and microstructures are characterized by Sievert-type adsorption apparatus, BET, SEM, TEM/STEM, EDX and EELS. Compared with pristine graphene, Mn decoration marginally increases the hydrogen adsorption capacity of graphene at room temperature and 4 MPa hydrogen pressure from 0.25 wt% to 0.36 wt%. On the other hand, the co-decoration of Mn and V increases the room temperature hydrogen storage capacity of graphene significantly to 1.81 wt% under 4 MPa hydrogen pressure, which is 1.56 wt% higher than the capacity of pristine graphene. The microstructures and valence states of the decorated Mn and Mn-V nanoparticles are investigated by TEM, EDX and EELS analyses, and strong interactions between the decorated nanoparticles and graphene are observed. Based on the results from structural analyses, potential enhancement mechanisms are suggested in terms of the catalytic effects of nanoparticles on graphene hydrogen adsorption. Given the relatively low cost of Mn and V metals compared to noble metals such as Pd, Pt and Au, these results demonstrate a low cost and effective way to significantly enhance the room temperature hydrogen adsorption properties of graphene for potential hydrogen storage applications.

The effects of 1,4-cyclohexanediol on frozen ram spermatozoa.

In order to improve the quality of frozen spermatozoa of Yunnan semi-fine wool sheep, 1, 4-cyclohexanediol (1, 4-CHD) as a synthetic ice blocker was used for cryopreservation of ram spermatozoa in this study. Briefly, following collection by electric stimulation, equilibration at 5℃ following dilution with the freezing extender, and pre-freezing in liquid nitrogen vapor, the ram spermatozoa were preserved in liquid nitrogen for one month. In addition, the effects of osmolarity of the diluting extenders used for evaluation of frozen spermatozoa quality were also assessed. The results indicated addition of 1, 4-CHD could not increase the motility of ram spermatozoa after cryopreservation and thawing. With the elevation of the concentrations of 1, 4-CHD, the motility and moving velocity of frozen ram spermatozoa showed a steady decrease. Additionally, the presence of 1, 4-CHD cannot increase the percentage of frozen spermatozoa with intact acrosome and membrane. When the isotonic binding buffer was used to dilute the thawed spermatozoa, the percentage of cells labeled with propidium iodide (PI) after cryopreservation in the presence of 1, 4-CHD was significantly higher than that of spermatozoa frozen in the absence of 1, 4-CHD (P < 0.05). However, the percentage of frozen-thawed spermatozoa with exposed PS in the presence of 1, 4-CHD was significantly less than that of spermatozoa frozen in the absence of 1, 4-CHD (P < 0.01). When the basic extenders with an osmolarity of 404mOsm, 528mOsm, 648mOsm, or 853mOsm were used to dilute the frozen-thawed spermatozoa respectively, there is no significant difference between the four groups with respect to the moving velocity and membrane integrity (P > 0.05). In conclusion, the presence of 1, 4-CHD cannot improve the motility, moving velocity, acrosome staus, and membrane integrity of frozen ram spermatozoa. However, 1, 4-CHD may inhibit apoptosis caused by freezing and thawing.

Stability of lyophilized human platelets loaded with small molecule carbohydrates.

Long-term preservation of platelets is a great challenge for blood transfusion centers, due to the required narrow storage temperature arange (22 ± 2 degree C). Short shelf life and potential bacterial growth often lead to the shortage of high-quality platelets. Freeze-dried preservation is thus believed to be a potential solution for long-term platelet storage without losing the hemostasis function. Here we report a new platelet preservation method, which uses small molecule carbohydrates to extend storage time and to maintain platelet function. The activities of lyophilized platelets that were stabilized with small molecule carbohydrate (e.g., cell viability, mean platelet volume, activation characteristics, and aggregation kinetics) were maintained after storage of 30, 60, and 90 days at room temperature, 4 degree C, and -20 degree C. The recovery of freeze-dried platelets was 87 percent in comparison to fresh platelets. The mean platelet volume of rehydrated platelets increased (from 6.8 fl to 8.0 fl). About 40 percent of rehydrated platelets was in the early-activated stage (PCA-1 positive) and 30 percent was in the terminal-activated stage (CD62P positive). The cell viability was about 60 percent as measured with CMFDA vital probes. The aggregation rate of rehydrated platelets after 90-day storage was similar to fresh platelets stored at 22 degree C ± 2 degree C.

The Wistar Bonn Kobori rat, a unique animal model for autoimmune pancreatitis with extrapancreatic exocrinopathy.

The male Wistar Bonn/Kobori (WBN/Kob) rat is known to be a unique animal model for chronic pancreatitis with widely distributed fibrosis and degeneration of parenchyma because of the infiltration of lymphocytes. In this report, we show that female (but not male) rats develop dacryoadenitis at 3 months of age, and that both male and female WBN/Kob rats develop sialoadenitis, thyroiditis, sclerotic cholangitis and tubulointerstitial nephritis over 18 months of age. The infiltration of CD8+ cells and the deposits of tissue-specific IgG2b were observed in the injured pancreas and lachrymal glands. Furthermore, the number of regulatory T cells (defined as CD4+ Forkhead box P3+ cells) decreased in the periphery of both male and female WBN/Kob rats, suggesting that the onset of these diseases is attributable, at least, to the failure in the maintenance of peripheral immune tolerance. These features show clearly that WBN/Kob rats are a useful animal model for autoimmune pancreatitis and Sjøgren-like syndrome or multi-focal fibrosclerosis in humans. We also show that these autoimmune diseases can be prevented by a newly devised strategy of bone marrow transplantation (BMT) in which bone marrow cells are injected directly into the bone marrow cavity: intrabone marrow-BMT.

Sera from patients with toxic epidermal necrolysis contain autoantibodies to periplakin.

BACKGROUND: The pathophysiological mechanism of toxic epidermal necrolysis (TEN) with extensive bullae that is induced suddenly by drugs is not well understood. The individual patterns and distribution of the widespread mucocutaneous reactions of TEN often show striking similarities with those of paraneoplastic pemphigus (PNP), which is known to involve autoantibodies (aAbs) to members of the plakin family. OBJECTIVES: To investigate the existence of circulating aAbs to periplakin in the sera of patients with TEN. METHODS: The presence of circulating aAbs to periplakin was examined using immunoblotting, immunoabsorption and indirect immunofluorescence (IF) analyses. Recombinant protein expression was used to determine the interaction between periplakin and aAbs in the sera of patients with TEN. RESULTS: Indirect IF studies revealed circulating aAbs in the intercellular area in the epidermis. Interestingly, on rat bladder the staining pattern of the IgG deposits was similar to that observed in patients with PNP. Immunoblotting analysis of the epidermal extracts was used to identify the aAbs in the sera of patients with TEN. These contained circulating aAbs to a 190-kDa protein corresponding to periplakin. Recombinant periplakin and domains of periplakin were prepared in order to confirm the existence of aAbs to periplakin. Immunoblotting with these proteins demonstrated that the sera from patients with TEN reacted with each domain as well as with the full-length periplakin. CONCLUSIONS: We found that circulating aAbs in the sera of patients with TEN target periplakin. These aAbs might play a role in the pathogenesis of TEN as a humoral autoimmune mechanism.

Receptor of activated C kinase 1 (RACK1) is necessary for the 20-hydroxyecdysone-induced expression of the transcription factor CHR3 in the spruce budworm Choristoneura fumiferana.

To initiate moulting and metamorphosis, 20-hydroxyecdysone (20E) binds to its nuclear receptors and the ligand-receptor complex then mediates changes in gene expression. Phosphorylation of the receptors is required for their function. The intracellular signal transduction pathway that is involved in receptor phosphorylation remains elusive. This study provides evidence that the receptor of activated C kinase 1 (RACK1) and protein kinase C (PKC) signal transduction cascade is involved in the 20E-induced expression of the moult-associated transcription factor CHR3. A cDNA clone encoding a receptor of activated C kinase 1 was isolated from Choristoneura fumiferana (CfRACK1). This single copy gene coded a 36 kDa protein and was expressed ubiquitously in all of the developmental stages and the tissues tested, including the midgut, epidermis, fat body, head, Malpighian tubules, ovary and testis of larvae. High levels of the transcripts were also detected in a midgut-derived CF-203 cell line. We noticed that the green fluorescence protein-fused CfRACK1 protein was distributed in the cytosol surrounding the nuclei in stably transformed cells. Interference of CfRACK1 mRNA suppressed the 20E-induced expression of the transcription factor CHR3. Dequalinium-14; 1,1'-decamethylenebis-4-aminoquinaldinium diiodide (DECA), an inhibitor of RACK1 binding to protein kinase C, blocked the 20E-induced expression of CHR3 and accumulation of the ecdysone receptor (EcR) in the nuclei. All of these data together suggest that 20E-induced expression of CHR3 may involve phosphorylation of the ecdysone receptor component through the PKC/RACK1 signal transduction cascade, which facilitates the import of the receptor into the nuclei of cells.

Use of RNAi technology to confer enhanced resistance to BmNPV on transgenic silkworms.

dsRNA is a powerful tool for gene-specific silencing in plants and animals. In this study, we examined the use of gene silencing in generating transgenic silkworms resistant to the Bombyx mori nucleopolyhedrovirus (BmNPV). Using a transposon piggyBac system, we first generated BmN cells (rBmN-lef1), which carried artificial genes designed for expressing dsRNAs with sequences of the essential viral gene lef-1. NPV DNA microarray analysis revealed that the accumulation of lef-1 mRNA was successfully inhibited in rBmN-lef1 infected with BmNPV. The virus titer in the culture medium of rBmN-lef1 at 48 hr post-infection (h.p.i.) was 50% of that of the control cells. Moderate BmNPV-resistance caused by transgenesis of the artificial dsRNA-expressing gene was confirmed in the transgenic silkworms. Virus production was reduced in transgenic silkworms relative to controls up to 96 hrs after viral inoculation. Although complete protection was not achieved and the transgenic larvae ultimately died, this is the first report to show the use of RNAi in confering enhanced viral resistance on transgenic animals.

Induction of the white egg 3 mutant phenotype by injection of the double-stranded RNA of the silkworm white gene.

Injection of double-stranded RNA (dsRNA) corresponding to the silkworm white gene (Bmwh3) into preblastoderm eggs of the wild-type silkworm induced phenotypes similar to those observed with mutants of the white egg 3 locus (10-19.6). The induced phenotypes were characterized by the presence of white eggs and translucent larval skin. Northern analysis showed that the expression of the endogenous Bmwh3 gene in the injected embryos was distinctly depressed. Furthermore, the injection of the GFP dsRNA inhibited the expression of the GFP gene from a plasmid co-injected with the dsRNA but did not depress the expression of the Bmwh3 gene. These findings demonstrate that sequence-specific RNA interference occurred in the silkworm. We conclude from the results that the RNA interference can be applied as a tool for the analysis of the gene function in the lepidopteran insects.

Characterization of the kynurenine 3-monooxygenase gene corresponding to the white egg 1 mutant in the silkworm Bombyx mori.

Kynurenine 3-monooxygenase (KMO, EC 1.14.13.9), which catalyzes the oxidation of kynurenine to 3-hydroxykynurenine, is involved in the synthesis of ommochrome pigments in insects. A silkworm mutant, white egg 1 ( w-1), has been shown to be deficient in this enzyme activity. The mutant is characterized morphologically by its white eyes and the fact that the females lay white eggs. To analyze the relationship between the KMO gene and the mutation, we first determined the entire sequence of a full-length 2.0-kb cDNA and examined its expression pattern in the wild type. The cDNA sequence contains one ORF encoding a polypeptide of 456 amino acids, and transcripts were detected in the larval Malpighian tubules and the pupal ovaries, but not in other tissues. Southern analysis and nucleotide sequencing showed that the KMO gene is present in a single copy and consists of ten exons distributed over a 16-kb region. Comparison of the transcripts between the wild type and mutant silkworms showed that the wild type expressed a single transcript, whereas the mutant exhibited markedly reduced amounts of two transcripts with sizes of 2.0 kb and 1.8 kb. Nucleotide sequence analysis of these mutant transcripts indicated that sequences corresponding to the ninth and tenth exons were missing. Inverse PCR and Southern analysis of the mutant gene demonstrated that the corresponding genomic region was deleted in the w-1 mutant.

Isolation and expression of an ecdysteroid-inducible neutral endopeptidase 24.11-like gene in wing discs of Bombyx mori.

In the process of comparison of two cDNA libraries (W0, W2), we isolated a clone from the wing discs of Bombyx mori encoding a putative neutral endopeptidase 24.11-like gene. The predicted open reading frame encoded 772 amino acid residues, having about 53% identity with Drosophila GH07643, 36% with rat NEP, and 34% with rat ECE. This is the first NEP gene isolated in invertebrate. A 3.6-kb transcript was found to accumulate in the wing disc according to the increase of ecdysteroid titer during metamorphosis. Accumulation of the transcript was induced in wing discs with 20-hydroxyecdysone about 20h after incubation, which was inhibited by cycloheximide. This gene is ecdysone-inducible, appears to encode a functional protein, and may function during wing metamorphosis.

Important causes of hypoglycaemia in patients with diabetes on peritoneal dialysis.

AIM: Diabetes is now the commonest cause of end-stage renal failure, so there are many diabetic patients receiving dialysis therapy. There are several important ways in which dialysis practice can impinge unfavourably on glucose control. This study focuses on the interaction between maltose-derived metabolites in a new peritoneal dialysis fluid and blood glucose measurements using reagent sticks that depend on the glucose dehydrogenase method. CASE REPORT: We report the cases of three patients, with insulin-treated diabetes and end-stage renal disease treated with peritoneal dialysis, who experienced symptomatic hypoglycaemia with inaccurate glucose readings on reagent strips when converted to icodextrin. CONCLUSION: Careful teamwork between diabetes and renal physicians and specialist nurses is highly desirable to achieve good glucose control in a group of patients at particular risk of microvascular and macrovascular complications.

The effect of T-2 toxin on IL-1beta and IL-6 secretion in human fetal chondrocytes.

The effects of T-2 toxin on IL-1beta and IL-6 secretion in human fetal chondrocytes in vitro were investigated. The evaluation is realised on primary monolayer culture of human fetal epiphyseal chondrocytes with or without PMA stimulation. The levels of supernatant IL-1beta and IL-6 were analyzed by ELISA. As compared with their respective controls, we observed a significant increase of IL-Ibeta and IL-6 in supernatants of chondrocytes cultivated for 24 h with T-2 at 8 ng/ml after PMA stimulation; in the absence of PMA, IL-Ibeta was increased alone after 48 h. The results demonstrated that T-2 toxin could superinduce IL-1beta and IL-6 secretion in chondrocytes. All these data suggested that superinduction of cytokines might be one of the key mechanisms of chondrocyte injuries by T-2 toxin.

Mass isolation of cuticle protein cDNAs from wing discs of Bombyx mori and their characterizations.

Multiple cloning of cuticle protein genes was performed by sequencing of cDNAs randomly selected from a cDNA library of wing discs just before pupation, and nine different cuticular protein genes were identified. Thirty-one clones of a cuticle protein gene were identified from the 1050 randomly sequenced clones; about 3% were cuticle protein genes in the W3-stage wing disc cDNA library. The sequence diversity of the deduced amino acid sequences of isolated Bombyx cuticle genes was examined along with the expression profiles. The deduced amino acid sequences of the nine cuticle protein genes contained a putative signal peptide at the N-terminal region and a very conserved hydrophilic region known as the R and R motif. The developmental expression of cuticle genes was classified into two types: pupation (five clones were expressed only around pupation) and pupation and mid-pupal (four clones were expressed around this stage). All the isolated genes were expressed in the head, thoracic, and abdominal regions of the epidermis at different levels around pupation, but no expression was observed in the epidermis at the fourth molting stage.

Transfusion practice in elective orthopaedic surgery.

. The transfusion requirements of 2233 patients who underwent total hip or knee joint arthroplasty procedures at nine Canadian hospitals during 1995-1996 were evaluated. Although 64% of patients were eligible for participation in an autologous blood donation (ABD) programme, only 8% predonated blood. Patients who were eligible for ABD were younger (62 years vs. 70 years) and had fewer medical illnesses (18% vs. 44%) than those who did not predonate. The rate of allogeneic transfusion was 9.0% (95% confidence interval 4.9-13.1%) in patients who predonated as compared with 24.1% (95% confidence interval 22.2-25.9%) in those who did not. Risk factors for the occurrence of an allogeneic transfusion were type of procedure (primary or revision hip arthroplasty), lower baseline haemoglobin, lower body weight, older age and presence of rheumatoid arthritis (P < 0.001). Only patients without risk factors were predicted to have a less than 10% risk of receiving an allogeneic transfusion. Use of preventive strategies was minimal. Two models designed to predict the occurrence of an allogeneic transfusion were evaluated. If allogeneic transfusion rates are to be reduced, eligible patients should be encouraged to participate in ABD programmes. For patients who are ineligible, other preventative strategies should be introduced.

Isolation and expression of the ecdysteroid-inducible angiotensin-converting enzyme-related gene in wing discs of Bombyx mori.

We isolated a clone encoding a putative angiotensin-converting enzyme-related gene from the wing disc cDNA library of the silkworm, Bombyx mori (refer to as BmAcer). The predicted open reading frame encoded 648 amino acids with about 50% identities with the Drosophila melanogaster angiotensin-converting enzyme Ance and Acer. Northern analysis identified a 2.2-kilobase mRNA which was abundant in wing discs two days after the beginning of wandering. An accumulation of the transcript was observed approximately 2 h after 20-hydroxyecdysone (20E) exposure in vitro and was blocked slightly by a protein synthetic inhibitor. These data suggest that the transcription of the BmAcer gene is directly 20E-inducible.