[Molecular epidemiology of Listeria monocytogenes isolated from ready-to-eat food in 2017 in China].

Objective: To analyze the molecular characteristics of Listeria monocytogenes strains from ready-to eat food in China. Methods: A total of 239 Listeria monocytogenes strains isolated from ready-to-eat food in 2017, all strains underwent whole-genome sequencing (WGS) , and comparisons uncovered population structure derived from lineages, clonal complex, serogroups, antimicrobial susceptibility and virulence, which were inferred in silico from the WGS data. Core genome multilocus sequence typing was used to subtype isolates. Results: All strains were categorized into three different lineages, lineage Ⅱ was the predominant types in food, and IIa was the main serogroups. CC8, CC101 and CC87 were the first three prevalent CCs among 23 detected CCs, accounting for 49.4%. Only 4.6% (11 isolates) of tested strains harbored antibiotic resistance genes, which were mostly trimethoprim genes (7 isolates, 2.9%). All strains were positive for LIPI-1, and only a part of strains harbored LIPI-3 and LIPI-4, accounting for 13.8% (33 isolates) and 14.2% (34 isolates), respectively. ST619 carried both LIPI-3 and LIPI-4. 51.5% (123 isolates) of strains carried SSI-1, and all CC121 strains harbored SSI-2. Different lineages, serogroups and CCs can be separated obviously through cgMLST analysis, and 24 sublineages were highly concordant with CCs. Conclusion: Ⅱa was the main serogroups in ready-to-eat food isolates in China; CC8, CC101 and CC87 were the prevalent CCs, and CC87 isolates was hypervirulent isolates, cgMLST method can be adopted for prospective foodborne disease surveillance and outbreaks detection.

Effects of culture conditions on spore types of Clonostachys rosea 67-1 in submerged fermentation.

UNLABELLED: The promising biocontrol isolate Clonostachys rosea 67-1 was investigated to clarify the effects of culture conditions on chlamydospore production in submerged fermentation. Culture conditions significantly affected both performance and types of C. rosea sporulation. C. rosea 67-1 was hard to generate chlamydospores under conventional conditions. However, the proportion of resistant spores increased to 17·4 and 15·5% in PD and rice meal media, respectively, in 8 days. Chlamydospore productivity was boosted (>threefold) with the addition of 50-200 mg l(-1) CuSO4 . The pH of the medium played a vital role in 67-1 sporulation. The percentage of chlamydospores decreased rapidly with increased pH (88·1% at pH 3·0 to 1·0% at pH 6·5). The optimal pH for conidia production was 6·0-6·5, at which chlamydospore forming was strongly inhibited. Regulating pH during fermentation contributed to improving output and proportion of resistant spores. When 67-1 was inoculated into broth with an initial pH of 6·5, followed by adjustment to pH 3·5 after 48 h, the number of chlamydospores reached 1·1 × 10(8) ml(-1). The impact of temperature and rotational speed was also analysed; an ultimate capacity of chlamydospores was achieved at 30°C and the speed above 120 rev min(-1) (P < 0·05). SIGNIFICANCE AND IMPACT OF THE STUDY: Clonostachys rosea is one of the most promising biocontrol agents in countering many plant fungal diseases. However, large-scale production and commercialization are hampered by the lack of understanding of the impacts of culture conditions on performance and types of C. rosea sporulation and subsequently inadequate research on the techniques for chlamydospore production. In addressing these concerns, this study provides a unique insight into the manipulation of C. rosea sporulation and chlamydospore fermentation of the biocontrol fungus.

The relationship among renal injury, changed activity of renal 1-alpha hydroxylase and bone loss in elderly rats with insulin resistance or Type 2 diabetes mellitus.

BACKGROUND AND OBJECTIVE: Chronic kidney disease can lead to a decrease in active vitamin D [1,25-(OH)2D], which may be reversed by 1-alpha hydroxyvitamin D [1-alpha(OH)D]. Renal 1-alpha hydroxylase, expressed in renal tubular epithelial cells, is a key enzyme in the synthesis of 1,25-(OH)2D. 1,25- (OH)2D plays an important role in the regulation of calcium and phosphate metabolism, and its deficiency can result in osteoporosis. Type 2 diabetes mellitus (T2DM) and insulin resistance (IR) are associated with renal injury, decrease in 1,25-(OH)2D and bone loss. The study aimed to explore the relationship among renal injury, decrease in 1,25-(OH)2D and bone loss in the presence of IR or T2DM, as well as the role of renal 1-alpha hydroxylase in the process. MATERIALS AND METHODS: Fifty 18-month-old male Wistar rats were randomized into 5 groups: normal control group (Group N), IR group (Group I), T2DM group (Group D), No.1 treatment group (Group T1), and No.2 treatment group (Group T2), 10 in each group. High-fat diet was administered to induce IR, while high-fat diet and low-dose streptozotocin were jointly applied to induce T2DM. Rats in Groups T1 and T2 were treated with vitamin D and 1-alpha(OH)D, respectively. At week 12, IR was determined by the use of euglycemic insulin clamp technique for rats in each group, and then glucose infusion rate (GIR) was calculated. Meanwhile, urinary albumin (UA), serum 25-(OH)D and 1,25-(OH)2D levels were determined by radioimmunoassay. After the rats were sacrificed, bone mineral density (BMD) in femoral bone and lumbar vertebrae was measured by the use of dual energy X-ray absorption. RESULTS: The GIR in Group N was significantly higher than that of the other 4 groups (p<0.01). Compared with Groups N (p<0.01) or I (p<0.05), the UA levels in Groups D, T1, and T2 were obviously higher. The UA level in Group I was higher than that of Group N, but the difference was not significant (p>0.05). In Groups D and I, the UA levels showed a negative correlation with GIR. No significant difference was observed in the levels of 25-hydroxyvitamin D [25-(OH)D]. The levels of 1,25-(OH)2D in Groups D and T1 were markedly lower than that of Groups N or T2 (p<0.01). The 1,25-(OH)2D level in Group I was lower than that of Group N (p<0.05), but higher than that of Group D (p<0.01). The 1,25-(OH)2D level in Group T2 was nearly equivalent to that of Group N. In Groups D and I, the levels of 1,25-(OH)2D were negatively correlated with UA, and positively correlated with GIR. The BMD levels in lumbar vertebrae or femoral bone in Groups D and T1 were similar, but both were lower than that of Groups T2 (p<0.05) and N (p<0.01). The BMD levels were lower in Groups I and T2 compared with that of Group N (p<0.05), but higher than that of Groups D and T1 (p<0.05). The BMD levels in lumbar vertebrae or femoral bone in Groups I and D were positively correlated with GIR. The BMD level in lumbar vertebrae or femoral bone in Group D showed negative correlation with UA. CONCLUSION: In elderly rats with T2DM or IR, renal injury may cause decreased activity of renal 1-alpha hydroxylase, which may result in bone loss and disturbance in VD metabolism, mainly manifesting as a significant reduction in the 1,25-(OH)2D level.

The relationship between renal injury and change in vitamin D metabolism in aged rats with insulin resistance or type 2 diabetes mellitus.

Insulin resistance (IR), IR treated with vitamin D, IR treated with 1alpha-hydroxyvitamin D (1alpha-(OH)D), type 2 diabetes mellitus (T2DM), T2DM treated with vitamin D and T2DM treated with 1alpha-(OH)D were studied in animal models using aged Wistar rats. Glucose infusion rates and levels of urinary albumin (UA), serum 25-hydroxyvitamin D (25-(OH)D) and 1,25-dihydroxyvitamin D (1,25(OH)2D) were measured. T2DM rats had higher UA than IR or normal rats, and levels of 25-(OH)D in all models weresimilar. IR rats had higher 1,25-(OH)2D levels than T2DM rats, and had lower 1,25-(OH)2D levels than normal rats. Treating IR or T2DM rats with vitamin D had no effect on 25-(OH)D or 1,25-(OH)2D. Administration of 1alpha-(OH)D significantly increased 1,25-(OH)2D in IR rats to above-normal levels, and significantly increased 1,25-(OH)2D in T2DM rats to normal levels. In IR or T2DM, abnormal vitamin D metabolism is characterized by 1,25-(OH)2D deficiency and is related to renal injury.

[Progress in knowledge-based X-ray fluorescence spectrometry].

The review focuses on the expert systems and knowledge engineering in X-ray fluorescence spectrometry. It includes mainly a knowledge-controlled strategy combining the scan-based method with the fixed channel measurements, a XRF interpretation system of spectra with fuzzy logic and pattern recognition, and an expert system for qualitative interpretation of XRF spectra using a certainty factor. In the review, a series of the studies of exploring the knowledge engineering system in XRF are also included, which consists of four parts, i.e. spectra identification, pattern recognition with decision-making, quantitative determination combined with the theoretical alpha coefficients and neural networks, and XRF analysis without standards.

Inhibition of Bcr serine kinase by tyrosine phosphorylation.

The first exon of the BCR gene encodes a new serine/threonine protein kinase. Abnormal fusion of the BCR and ABL genes, resulting from the formation of the Philadelphia chromosome (Ph), is the hallmark of Ph-positive leukemia. We have previously demonstrated that the Bcr protein is tyrosine phosphorylated within first-exon sequences by the Bcr-Abl oncoprotein. Here we report that in addition to tyrose 177 (Y-177), Y-360 and Y283 are phosphorylated in Bcr-Abl proteins in vitro. Moreover, Bcr tyrosine 360 is phosphorylated in vivo within both Bcr-Abl and Bcr. Bcr mutant Y177F had a greatly reduced ability to transphosphorylate casein and histone H1, whereas Bcr mutants Y177F and Y283F had wild-type activities. In contrast, the Y360F mutation had little effect on Bcr's autophosphorylation activity. Tyrosine-phosphorylated Bcr, phosphorylated in vitro by Bcr-Abl, was greatly inhibited in its serine/threonine kinase activity, impairing both auto- and transkinase activities of Bcr. Similarly, the isolation of Bcr from cells expressing Bcr-Abl under conditions that preserve phosphotyrosine residues also reduced Bcr's kinase activity. These results indicate that tyrosine 360 of Bcr is critical for the transphosphorylation activity of Bcr and that in Ph-positive leukemia, Bcr serine/threonine kinase activity is seriously impaired.

Rotavirus infection of the oropharynx and respiratory tract in young children.

Oropharyngeal aspirates were obtained from 89 infants hospitalized with respiratory illnesses accompanied or not by diarrhea and 33 control patients without the diseases. Rotavirus was detected from 25 of these patients by immunocytology, isolation of the virus in cultures of MA104 cells, or both. None of the control patients gave a positive result. The infection involves squamous cells and globlet cells probably originating from the oropharynx, and ciliated columnar epithelial cells from the respiratory tract. The virus from 2 specimens was propagated by repeatedly passaging in the cultures and found to have characteristic morphology of rotavirus. The electrophoretic patterns of the viral RNA extracted from them are closely similar to those obtained with the rotavirus genome extracted from the stool of the same patients. Repeated stool specimens were also obtained, and sera were paired from some of these subjects. All but one of the patients who gave a positive virology for their aspirates also showed a significant rise in the titres of common group A rotavirus antibody, neutralizing antibody against one or more of serotypes of rotavirus, or both. Patients who excreted rotavirus in their stools were younger and had significantly lower titres of rotavirus antibodies in their acute sera, than those who shedded the virus in the oropharynx but did not excrete the virus in repeated stool specimens. The prevalence of rotavirus in the oropharyngeal aspirates from these patients surpassed that of adenovirus, respiratory syncytial virus, influenza virus, and herpes simplex virus combined.

Development of neutralizing antibodies and group A common antibodies against natural infections with human rotavirus.

We determined the levels of group A common and neutralizing antibodies against human rotavirus in paired serum specimens obtained from 38 infants within 12 days of the onset of diarrhea. Thirty of the infants excreted rotavirus in stools, and eight did not. Nine patients (30%) with rotavirus diarrhea and seven patients (88%) with diarrhea due to other causes had detectable levels (greater than or equal to 1: 80) of immunoglobulin (IgG) common antibodies in acute-phase sera. All the patients with rotavirus diarrhea showed at least fourfold rises in titers of IgG or IgM common antibodies or both, while only two control patients showed significant rises in either IgG or IgM common antibodies in their convalescent-phase sera. Of the 19 patients excreting "short" electropherotypes of rotavirus, 18 showed at least fourfold rises in titers of neutralizing antibodies against serotype 2 human rotavirus but not against serotype 1, 3, or 4. Nine of the ten patients excreting "long" electropherotypes showed significant rises in neutralizing antibodies against serotype 3, and the other patient showed a significant rise in neutralizing antibodies against serotype 1. One patient excreted long and short electropherotypes simultaneously, and he also showed a significant rise in neutralizing antibodies against serotype 2 and 3 viruses. The control patients with diarrhea did not show significant changes in titers of antibodies against any of the serotypes. These results demonstrated that the neutralizing antibody response within 2 weeks after clinical onset is specific for the infecting serotype of rotavirus.