[HbA1c comparison and diagnostic efficacy analysis of multi center different glycosylated hemoglobin detection systems].

Objective: Compare and analyze the results of the domestic Lanyi AH600 glycated hemoglobin analyzer and other different detection systems to understand the comparability of the detection results of different detectors, and establish the best cut point of Lanyi AH600 determination of haemoglobin A1c (HbA1c) in the diagnosis of diabetes. Methods: Multi center cohort study was adopted. The clinical laboratory departments of 18 medical institutions independently collected test samples from their respective hospitals from March to April 2022, and independently completed comparative analysis of the evaluated instrument (Lanyi AH600) and the reference instrument HbA1c. The reference instruments include four different brands of glycosylated hemoglobin meters, including Arkray, Bio-Rad, DOSOH, and Huizhong. Scatter plot was used to calculate the correlation between the results of different detection systems, and the regression equation was calculated. The consistency analysis between the results of different detection systems was evaluated by Bland Altman method. Consistency judgment principles: (1) When the 95% limits of agreement (95% LoA) of the measurement difference was within 0.4% HbA1c and the measurement score was≥80 points, the comparison consistency was good; (2) When the measurement difference of 95% LoA exceeded 0.4% HbA1c, and the measurement score was≥80 points, the comparison consistency was relatively good; (3) The measurement score was less than 80 points, the comparison consistency was poor. The difference between the results of different detection systems was tested by paired sample T test or Wilcoxon paired sign rank sum test; The best cut-off point of diabetes was analyzed by receiver operating characteristic curve (ROC). Results: The correlation coefficient R2 of results between Lanyi AH600 and the reference instrument in 16 hospitals is≥0.99; The Bland Altman consistency analysis showed that the difference of 95% LoA in Nanjing Maternity and Child Health Care Hospital in Jiangsu Province (reference instrument: Arkray HA8180) was -0.486%-0.325%, and the measurement score was 94.6 points (473/500); The difference of 95% LoA in the Tibetan Traditional Medical Hospital of TAR (reference instrument: Bio-Rad Variant II) was -0.727%-0.612%, and the measurement score was 89.8 points; The difference of 95% LoA in the People's Hospital of Chongqing Liang Jiang New Area (reference instrument: Huizhong MQ-2000PT) was -0.231%-0.461%, and the measurement score was 96.6 points; The difference of 95% LoA in the Taihe Hospital of traditional Chinese Medicine in Anhui Province (reference instrument: Huizhong MQ-2000PT) was -0.469%-0.479%, and the measurement score was 91.9 points. The other 14 hospitals, Lanyi AH600, were compared with 4 reference instrument brands, the difference of 95% LoA was less than 0.4% HbA1c, and the scores were all greater than 95 points. The results of paired sample T test or Wilcoxon paired sign rank sum test showed that there was no statistically significant difference between Lanyi AH600 and the reference instrument Arkray HA8180 (Z=1.665,P=0.096), with no statistical difference. The mean difference between the measured values of the two instruments was 0.004%. The comparison data of Lanyi AH600 and the reference instrument of all other institutions had significant differences (all P<0.001), however, it was necessary to consider whether it was within the clinical acceptable range in combination with the results of the Bland-Altman consistency analysis. The ROC curve of HbA1c detected by Lanyi AH600 in 985 patients with diabetes and 3 423 patients with non-diabetes was analyzed, the area under curve (AUC) was 0.877, the standard error was 0.007, and the 95% confidence interval 95%CI was (0.864, 0.891), which was statistically significant (P<0.001). The maximum value of Youden index was 0.634, and the corresponding HbA1c cut point was 6.235%. The sensitivity and specificity of diabetes diagnosis were 76.2% and 87.2%, respectively. Conclusion: Among the hospitals and instruments currently included in this study, among these four hospitals included Nanjing Maternity and Child Health Care Hospital in Jiangsu Province (reference instrument: Arkray HA8180), Tibetan Traditional Medical Hospital of TAR (reference instrument: Bio-Rad Variant Ⅱ), the People's Hospital of Chongqing Liang Jiang New Area (reference instrument: Huizhong MQ-2000PT), and the Taihe Hospital of traditional Chinese Medicine in Anhui Province (reference instrument: Huizhong MQ-2000PT), the comparison between Lanyi AH600 and the reference instruments showed relatively good consistency, while the other 14 hospitals involved four different brands of reference instruments: Arkray, Bio-Rad, DOSOH, and Huizhong, Lanyi AH600 had good consistency with its comparison. The best cut point of the domestic Lanyi AH600 for detecting HbA1c in the diagnosis of diabetes is 6.235%.

Endophytic Bacillus subtilis P10 from Prunus cerasifera as a biocontrol agent against tomato Verticillium wilt / Bacillus subtilis P10 endofítico de Prunus cerasifera como agente de biocontrole contra murcha de Verticillium de tomate

Abstract Endophytic bacteria serve key roles in the maintenance of plant health and growth. Few studies to date, however, have explored the antagonistic and plant growth-promoting (PGP) properties of Prunus cerasifera endophytes. To that end, we isolated endophytic bacteria from P. cerasifera tissue samples and used a dual culture plate assay to screen these microbes for antagonistic activity against Verticillium dahliae, Botryosphaeria dothidea, Fusarium oxysporum, F. graminearum, and F. moniliforme. Of the 36 strains of isolated bacteria, four (strains P1, P10, P16, and P20) exhibited antagonistic effects against all five model pathogens, and the P10 strain exhibited the strongest antagonistic to five pathogens. This P10 strain was then characterized in-depth via phenotypic assessments, physiological analyses, and 16s rDNA sequencing, revealing it to be a strain of Bacillus subtilis. Application of a P10 cell suspension (1×108 CFU/mL) significantly enhanced the seed germination and seedling growth of tomato in a greenhouse setting. This P10 strain further significantly suppressed tomato Verticillium wilt with much lower disease incidence and disease index scores being observed following P10 treatment relative to untreated plants in pot-based experiments. Tomato plants that had been treated with strain P10 also enhanced defense-related enzymes, peroxidase, superoxide dismutase, and catalase activity upon V. dahliae challenge relative to plants that had not been treated with this endophytic bacterium. The results revealed that the P10 bacterial strain has potential value as a biocontrol agent for use in the prevention of tomato Verticillium wilt.

Resumo As bactérias endofíticas desempenham papel fundamental na manutenção da saúde e do crescimento das plantas. Poucos estudos até o momento, no entanto, exploraram as propriedades antagônicas e promotoras de crescimento de plantas (PGP) de endófitos de Prunus cerasifera. Para esse fim, isolamos bactérias endofíticas de amostras de tecido de P. cerasifera e usamos um ensaio de placa de cultura dupla para rastrear esses micróbios quanto à atividade antagonista contra Verticillium dahliae, Botryosphaeria dothidea, Fusarium oxysporum, F. graminearum e F. moniliforme. Das 36 cepas de bactérias isoladas, quatro (cepas P1, P10, P16 e P20) exibiram efeitos antagônicos contra todos os cinco patógenos modelo, e a cepa P10 exibiu o antagonista mais forte para cinco patógenos. Essa cepa P10 foi então caracterizada em profundidade por meio de avaliações fenotípicas, análises fisiológicas e sequenciamento de rDNA 16s, revelando ser uma cepa de Bacillus subtilis. A aplicação de uma suspensão de células P10 (1 × 108 UFC / mL) aumentou significativamente a germinação das sementes e o crescimento das mudas de tomate em casa de vegetação. Essa cepa P10 suprimiu ainda mais a murcha de Verticillium do tomate com incidência de doença muito menor e pontuações de índice de doença sendo observadas após o tratamento com P10 em relação a plantas não tratadas em experimentos baseados em vasos. As plantas de tomate que foram tratadas com a cepa P10 também aumentaram as enzimas relacionadas à defesa, peroxidase, superóxido dismutase e atividade da catalase após o desafio de V. dahliae em relação às plantas que não foram tratadas com essa bactéria endofítica. Os resultados revelaram que a cepa bacteriana P10 tem valor potencial como agente de biocontrole para uso na prevenção da murcha de Verticillium em tomate.

Endophytic Bacillus subtilis P10 from Prunus cerasifera as a biocontrol agent against tomato Verticillium wilt.

Endophytic bacteria serve key roles in the maintenance of plant health and growth. Few studies to date, however, have explored the antagonistic and plant growth-promoting (PGP) properties of Prunus cerasifera endophytes. To that end, we isolated endophytic bacteria from P. cerasifera tissue samples and used a dual culture plate assay to screen these microbes for antagonistic activity against Verticillium dahliae, Botryosphaeria dothidea, Fusarium oxysporum, F. graminearum, and F. moniliforme. Of the 36 strains of isolated bacteria, four (strains P1, P10, P16, and P20) exhibited antagonistic effects against all five model pathogens, and the P10 strain exhibited the strongest antagonistic to five pathogens. This P10 strain was then characterized in-depth via phenotypic assessments, physiological analyses, and 16s rDNA sequencing, revealing it to be a strain of Bacillus subtilis. Application of a P10 cell suspension (1×108 CFU/mL) significantly enhanced the seed germination and seedling growth of tomato in a greenhouse setting. This P10 strain further significantly suppressed tomato Verticillium wilt with much lower disease incidence and disease index scores being observed following P10 treatment relative to untreated plants in pot-based experiments. Tomato plants that had been treated with strain P10 also enhanced defense-related enzymes, peroxidase, superoxide dismutase, and catalase activity upon V. dahliae challenge relative to plants that had not been treated with this endophytic bacterium. The results revealed that the P10 bacterial strain has potential value as a biocontrol agent for use in the prevention of tomato Verticillium wilt.

First Report of Powdery Mildew Caused by Podosphaera fusca on Herba eupatorii in China.

Herba eupatorii, one of the most important Chinese medicinal herbs, belongs to the Asteraceae family. In June 2012, a previously unknown disease, tentatively identified as powdery mildew, was observed on H. eupatorii growing in Shangqiu, in eastern Henan Province, China. Symptoms began as white mycelium partially covering upper leaf surfaces; as the disease progressed, it spread to cover entire leaf surfaces. The infected leaves became yellow and necrotic at advanced stages of infection. Specimens consisting of infected leaves were maintained at the Plant-Microbe Interaction Laboratory at Shangqiu Normal University. Microscopic observations of the morphology of the fungus revealed oval primary conidia measuring 18 to 27 × 15 to 22 µm. A long unbranched germ tube that germinated laterally from the ends of conidia was observed in some samples. Conidiophores were cylindrical, simple unbranched, and composed of a basal cell with a swollen base and three to six barrel-shaped conidia formed in chains, measuring 112 to 180 × 9 to 12 µm. Mycelial appressoria were nipple-shaped. Chasmothecia were not observed in the collected samples. To verify the identity of the fungus, the internal transcribed spacer (ITS) rDNA was amplified with ITS1 and ITS4 primers (3) and sequenced. The sequences were deposited as GenBank Accession No. JX546297. Comparison with sequences in the GenBank database revealed that the ITS sequence was 100% homologous with the sequence of Podosphaera fusca on Calendula officinalis (AB525914) (2) and Syneilesis palmata (AB040349) (1). The ITS sequence analysis verified that the causal agent was P. fusca, which is reported to be a cosmopolitan powdery mildew fungus, parasitic on numerous plant species in the Asteraceae family. Koch's postulates were completed by inoculating healthy H. eupatorii plants with a conidial suspension (prepared in distilled water) of 105 conidia/ml collected from infected plants. Five plants were sprayed until the suspension ran off the leaves, while five additional plants were sprayed with distilled water as a control. Plants were maintained in a climate cell under the following conditions: day, 24°C, 16 h; night, 20°C, 8 h; 85% humidity. After 10 days, inoculated plants developed symptoms similar to those observed in the field, whereas control plants remained healthy. Further examination showed that the inoculated plants were infected by P. fusca. To our knowledge, this is the first report of P. fusca affecting H. eupatorii in China. Because there are no fungicides labeled for use on this plant, the appearance of powdery mildew caused by P. fusca could result in substantial production loss of H. eupatorii. References: (1) T. Hirata et al. Can. J. Bot. 78:1521, 2000. (2) S. Takamatsu et al. Persoonia 24:38, 2010. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, CA, 1990.

First Report of Powdery Mildew Caused by Golovinomyces cichoracearum on Sonchus asper in China.

Sonchus asper is an exotic annual herb belonging to the Asteraceae family. Crude extracts of S. asper leaves have antiphlogistic and antibacterial properties. In May 2011, severe infection of powdery mildew was observed on S. asper in Henan Province, a central area of China. Symptoms began as small, nearly circular white spots that later enlarged and coalesced. As the disease progressed, white mycelia covered the entire plant, including the inflorescence. The voucher specimens are kept at the Key Laboratory of Plant-Microbe Interaction, Shangqiu Normal University. Conidiophores were erect, 108 to 220 × 10 to 12 µm, and composed of a cylindrical foot cell followed by two to three short cells. Nipple-shaped appressoria were formed on ectophytic mycelia. Conidia were hyaline and cylindrical, without distinct fibrosin bodies, and ranging from 29 to 42 × 19 to 24 µm. Long, unbranched germ tubes were formed from the ends of the conidia. These morphological characteristics suggested that the powdery mildew fungus is the anamorph of Golovinomyces cichoracearum (1,2). Chasmothecia were not observed in the collected samples. To confirm the identification, the internal transcribed spacer (ITS) region of the rDNA was amplified with ITS1 and ITS4 primers (4) and sequenced. The ITS sequence obtained was assigned Accession No. JQ010848 in GenBank, and had 99% similarity to two ITS sequences from different G. cichoracearum accessions (HM449077 and AY739111). The ITS sequence analysis further verified that the causal agent of the powdery mildew on S. asper was G. cichoracearum. Pathogenicity was confirmed by inoculating five healthy plants of S. asper with a conidial suspension (105 conidia/ml) from the infected plants, with five noninoculated plants serving as control. Inoculated plants developed the original powdery mildew symptoms after 10 days, whereas control plants remained healthy. G. cichoracearum (synonym Erysiphe cichoracearum) is a cosmopolitan powdery mildew fungus, parasitic on numerous plant species in the Asteraceae family. It was reported to be infecting S. asper in Bulgaria (3). To our knowledge, this is the first report of G. cichoracearum infections on S. asper in China. S. asper is one of the most important Chinese herbal medicines. The sudden appearance of powdery mildew caused by G. cichoracearum on this plant could result in substantial production loss of S. asper unless fungicide applications are employed. References: (1) U. Braun. Beih. Nova Hedwigia 89:1, 1987. (2) R. T. A. Cook and U. Braun. Mycol. Res. 113:616, 2009. (3) G. Negrean and C. M. Denchev. Mycologia Balcanica, 2004, 1:63. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, CA, 1990.

Garlic accelerates red blood cell turnover and splenic erythropoietic gene expression in mice: evidence for erythropoietin-independent erythropoiesis.

Garlic (Allium sativum) has been valued in many cultures both for its health effects and as a culinary flavor enhancer. Garlic's chemical complexity is widely thought to be the source of its many health benefits, which include, but are not limited to, anti-platelet, procirculatory, anti-inflammatory, anti-apoptotic, neuro-protective, and anti-cancer effects. While a growing body of scientific evidence strongly upholds the herb's broad and potent capacity to influence health, the common mechanisms underlying these diverse effects remain disjointed and relatively poorly understood. We adopted a phenotype-driven approach to investigate the effects of garlic in a mouse model. We examined RBC indices and morphologies, spleen histochemistry, RBC half-lives and gene expression profiles, followed up by qPCR and immunoblot validation. The RBCs of garlic-fed mice register shorter half-lives than the control. But they have normal blood chemistry and RBC indices. Their spleens manifest increased heme oxygenase 1, higher levels of iron and bilirubin, and presumably higher CO, a pleiotropic gasotransmitter. Heat shock genes and those critical for erythropoiesis are elevated in spleens but not in bone marrow. The garlic-fed mice have lower plasma erythropoietin than the controls, however. Chronic exposure to CO of mice on garlic-free diet was sufficient to cause increased RBC indices but again with a lower plasma erythropoietin level than air-treated controls. Furthermore, dietary garlic supplementation and CO treatment showed additive effects on reducing plasma erythropoietin levels in mice. Thus, garlic consumption not only causes increased energy demand from the faster RBC turnover but also increases the production of CO, which in turn stimulates splenic erythropoiesis by an erythropoietin-independent mechanism, thus completing the sequence of feedback regulation for RBC metabolism. Being a pleiotropic gasotransmitter, CO may be a second messenger for garlic's other physiological effects.

A direct spectrophotometric assay for peptide deformylase.

A direct UV-VIS spectrophotometric assay has been developed for peptide deformylase. This assay employs a novel class of peptide mimetics as deformylase substrates which, upon enzymatic removal of the N-terminal formyl group, rapidly release free thiols. The released thiols are quantitated using Ellman's reagent. A variety of peptide analogues that contain beta-thiaphenylalanine or beta-thiamethionine as the N-terminal residue were synthesized and found to be excellent substrates of the peptide deformylase from Escherichia coli (k(cat)/K(M) = 6.9 x 10(5) M(-1) s(-1) for the most reactive substrate). The deformylase reaction is conveniently monitored on a UV-VIS spectrophotometer in a continuous fashion. The versatility of the assay has been demonstrated by its application to kinetic characterization of the deformylase, pH profile studies, and enzyme inhibition assays. The assay can also be performed in an end-point fashion. The results demonstrate that this assay is a simple, highly sensitive, and rapid method to study kinetic properties of deformylases without the use of any coupling enzymes.

CA-MMP: a matrix metalloproteinase with a novel cysteine array, but without the classic cysteine switch.

A matrix metalloproteinase (MMP)-like gene was identified in mouse to contain a conserved MMP catalytic domain and an RRRR motif. It lacks a classic cysteine switch, but it possesses two novel motifs: a cysteine array (Cys-X(6)-Cys-X(8)-Cys-X(10)-Cys-X(3)-Cys-X(2)-Cys), and a novel Ig-fold. It is named CA-MMP after the distinct cysteine array motif, and little is known about its biochemical function. In an attempt to characterize CA-MMP activity, the full-length sequence was expressed in mammalian cells and its product found to be cell-associated without detectable secretion. In light of this unusual finding, a chimera combining the catalytic domain of CA-MMP with the prodomain of stromelysin-3 was constructed to express a fully active enzyme in mammalian cells. Purified CA-MMP catalytic domain expresses proteolytic activity against protein substrates in an MMP inhibitor sensitive fashion. Taken together, it is concluded that CA-MMP is an MMP with distinct structure, biochemical properties and evolutionary history that may define a new subclass of the MMP superfamily.

[Clinical study on effect of jinshisan on dissolution of extra-/ intrahepatic biliary stones].

In order to evaluate the role and mechanism of Jinshisan (JSS) in bile stone dissolution, 164 cases of extra-/ intrahepatic biliary stone patients were treated by a collaborated group, the therapeutic effect was observed by B-ultrasonography before and after treatment. Results showed: (1) Among the 164 cases, 26 cases (15.85%) were cured clinically; 73 (44.51%) were effective and 65 (39.63%) ineffective. (2) After treatment, in 84 patients of right hepatolith, the stone were diappeared in 10 cases, the size or number of stone decreased in 44 casess. In 34 patients with left hepatolith, the figure were 5 and 13 respectively. In 10 cases of choledocholith they were 10 and 10 respectively. While in 21 cases of multiple gallstone, only one case with stones of both sides disappeared, except 3 ineffective cases, others with stone decreased in size or number or disappeared unilaterally. (3) Gallston diappeared in 7 of 10 patients with size of stone 6 months. (5) Analysis of bile showed that JSS could increase the amount of bile acid, decrease the bilirubin, mucous and bicarbonate radical in bile.

[Effect of Gardenia jasminoides Ellis (GJE) on the blood flow of internal organs at the early stage of acute necrotizing hemorrhagic pancreatitis in rats].

Effect of GJE on the prevention and treatment of experimental acute pancreatitis was observed by means of testing pancreatic, hepatic, gastric and intestinal blood flow. The results show that the blood flow of internal organs that decreases significantly at the early stage of acute necrotizing hemorrhagic pancreatitis in rats can be picked up by GJE especially pancreatic.