Chaetoglobosin A Contributes to the Antagonistic Action of Chaetomium globosum Strain 61239 Toward the Apple Valsa Canker Pathogen Cytospora mali.

Apple Valsa canker (AVC) weakens apple trees and significantly reduces apple production in China and other East Asian countries. Thus far, very few AVC-targeting biocontrol resources have been described. Here, we present a thorough description of a fungal isolate (Chaetomium globosum, 61239) that has strong antagonistic action toward the AVC causal agent Cytospora mali. Potato dextrose broth culture filtrate of strain 61239 completely suppressed the mycelial growth of C. mali on potato dextrose agar, and strongly constrained the development of AVC lesions in in vitro infection assays. ultra-performance liquid chromatography (UPLC) and HPLC-MS/MS investigations supported the conclusion that strain 61239 produces chaetoglobosin A, an antimicrobial metabolite that inhibits C. mali. Using genome sequencing, we discovered a gene cluster in strain 61239 that may be responsible for chaetoglobosin A production. Two of the cluster's genes-cheA, a PKS-NRPS hybrid enzyme, and cheB, an enoyl reductase-were individually silenced, which significantly decreased chaetoglobosin A accumulation as well as the strain's antagonistic activity against C. mali. Together, the findings of our investigation illustrate the potential use of Chaetomium globosum for the management of AVC disease and emphasize the significant contribution of chaetoglobosin A to the antagonistic action of strain 61239.

First Report of Colletotrichum aenigma Causing Apple Glomerella Leaf Spot on the Granny Smith Cultivar in China.

Glomerella leaf spot (GLS) is a devastating fungal disease causing pre-mature defoliation on apple (Malus domestica). It was first reported in 1970s and since then has been reported in North America, South America and Asia. GLS disease is caused by Colletotrichum fungi and the pathogens are genetically diverse, encompassing at least nine species belonging to three species complexes (Velho et al. 2018). In August 2018, disease with sudden leaf necrosis symptom, typical of GLS symptom appearance, occurred in a Granny Smith orchard in Wugong county, China, over 70% tree leaves bared brown and necrotic lesions. Small leaf tissues (3-4 mm2) cut from lesion margins were surface sterilized for 30 s in 3% NaClO and 30 s in 75% ethanol, followed by rinsing three times in sterile water before transferring onto potato dextrose agar (PDA) plates (25 ± 2°C). Seven isolates were obtained, all producing round cottony colonies on PDA, being white to pale on the upper side and dark green on the reverse side. Conidia were single-celled, cylindrical and transparent (17.33 ± 1.29 × 5.11 ± 0.77 µm, n=50). Appressoria were single-celled, thick-walled, dark brown, oval or irregular shaped, sometimes lobed (9.07 ± 0.88 × 6.66 ± 0.33 µm, n=50). The morphological and cultural characteristics of the fungal isolates matched the descriptions of Colletotrichum aenigma (Weir et al. 2012). To confirm the species identity, genomic DNAs were isolated from two representative isolates (QSG1 and QSY1), and the internal transcribed spacer (ITS), actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), beta-tubulin (TUB2), manganese-superoxide dismutase (SOD2), chitin synthase (CHS), and calmodulin (CAL) regions were amplified by PCR using reported primers (Weir et al. 2012). The sequences were deposited in GenBank (Accession Nos. MT872061, MT873580, MT873581, MT888183, MT888185, MT888187 and MT888189 for QSG1, and MT834933, MT835166, MT873579, MT888182, MT888184, MT888186 and MT888188 for QSY1). BLASTn search against GenBank nr database showed that ITS sequences of the two strains showed high nucleotide identity (over 99%) to sequences derived from the Colletotrichum gloeosporioides species complex (CGSC). Further concatenated phylogenetic analysis with reported CGSC strains (Weir et al. 2012) placed QSG1 and QSY1 in the clade of C. aenigma. To fulfill Koch's postulates, field pathogenicity test was performed. The experiment was performed in an orchard located in Yangling in September 2020, with the daily average temperature ranging between 15 - 20 ℃. Healthy 'Granny Smith' leaves were surface sterilized with 70% alcohol and inoculated with conidial suspension (106 conidia/mL) using cotton swabs. For each isolate, 10 leaf inoculations were performed. Inoculation with distilled water served as a negative control. Inoculated leaves were covered with plastic bags to maintain high humidity and the bags were removed at 48 hours post inoculation (hpi). Conidium-inoculated leaves started to exhibit GLS-resembling necrotic lesions from 5 dpi onward. The lesion extent, however, varied among inoculated leaves, ranging from blurry, small-sized lesions to blight of entire leaf. In contrast with conidium inoculations, water-inoculated leaves remained asymptomatic until 14 dpi. Re-isolated fungi from the symptomic leaf tissues were identical to C. aenigma in morphological appearance. Taken together, this is the first report of GLS on 'Granny Smith' apple, and the first report of C. aenigma causing apple GLS in China. This information should provide important guideline for developing field control practices of GLS.

[Effect of renal sympathetic denervation on left ventricular hypertrophy and inflammatory factors in spontaneously hypertensive rats].

OBJECTIVE: To investigate the effect of renal sympathetic denervation on left ventricular hypertrophy and inflammatory factors in spontaneously hypertensive rats. METHODS: Thirty six spontaneously hypertensive rats (SHR) were divided into 3 groups with 12 animals in each group: SHR control group,operation group and sham operation group. Bilateral renal sympathectomy or sham operation were performed in operation and sham groups,respectively; another 12 WKY rats served as normal controls. The blood pressure and body weight were examined weekly. The animals were sacrificed at w1 and w6, rat hearts were collected and left ventricular mass index (LVMI) was calculated. The expression of TLR4,TNF-α and IL-6 in heart tissue were detected by immunohistochemistry and Western blot. RESULTS: The systolic blood pressure [(201.67 ± 11.09) mmHg compared with (140.0 ± 10.86)mmHg,P<0.05],diastolic blood pressure [(144.50 ± 10.48)mmHg compared with (78.50 ± 7.32)mmHg,P<0.05], LVMI (2.44 ± 0.05 compared with 1.93 ± 0.05,P<0.05),the expression of TLR4 (0.298 ± 0.004 compared with 0.126 ± 0.004, P<0.05), NF-κB (0.249 ± 0.006 compared with 0.195 ± 0.005, P<0.05),TNF-α(0.323 ± 0.004 compared with 0.146 ± 0.004,P <0.05), IL-6 (0.283 ± 0.005 compared with 0.207 ± 0.006, P<0.05) in SHR control group were significantly higher than those in WKY group. Compared to sham operation group,the systolic blood pressure (157.30 ± 9.35 compared with 197.30 ± 11.5, P<0.05),diastolic blood pressure (112.50 ± 6.25 compared with 146.80 ± 7.6, P<0.05),LVMI (2.32 ± 0.04 compared with 2.57 ± 0.09, P<0.05, TLR4 (0.198 ± 0.006 compared with 0.317 ± 0.008, P<0.05), NF-κB (0.208 ± 0.006 compared with 0.332 ± 0.007, P<0.05), TNF-α(0.27 ± 0.009 compared with 0.375 ± 0.004,P<0.05), IL-6 (0.218 ± 0.004 compared with 0.376 ± 0.009, P<0.05) in operation group were all decreased at w1 after sympathectomy. Six weeks after the operation,there were no significant differences in systolic blood pressure (197.50 ± 12.13 compared with 208.83 ± 10.23,P>0.05) and diastolic blood pressure (150.33 ± 7.74 compared with 151.50 ± 8.22, P>0.05) between denervated and sham-operated SHRs; however,the LVMI (2.46 ± 0.07 compared with 2.81 ± 0.05,P<0.05) and the expression of TLR4(0.301 ± 0.009 compared with 0.567 ± 0.006, P<0.05), NF-κB (0.251 ± 0.004 compared with 0.476 ± 0.009,P<0.05),TNF-α(0.324 ± 0.005 compared with 0.535 ± 0.006, P<0.05,IL-6 (0.285 ± 0.009 compared with 0.549 ± 0.007, P<0.05) in operation group were still significantly lower than those in sham operation group. CONCLUSION: Renal sympathetic denervation can significantly delay the progression of LVH in SHR, which may associated with lowering blood pressure and decreasing expression of TLR4, NF-κB,TNF-α, IL-6 in myocardial tissue.

Impact of renal denervation on renalase expression in adult rats with spontaneous hypertension.

The aim of this study was to investigate the impact of renal denervation on the blood pressure, plasma renalase content and expression of renalase and tyrosine hydroxylase (TH) in the kidney of spontaneous hypertensive (SH) rats and to explore the mechanism of renal denervation involved in lowering blood pressure. SH rats (n=48) were randomly assigned to baseline, surgery (renal denervation), sham and control groups. WKY rats matched in age (n=12) served as the baseline control group. All rats were housed until they were 12 weeks old. The rats in the baseline group and the WKY group rats were sacrificed, and blood and kidney were collected for examination. In the renal denervation, sham and control groups, the blood pressure was continuously monitored. One and six weeks after renal denervation, 6 rats in each group were sacrificed, and blood and kidney were collected for examination. ELISA was employed to measure the plasma renalase, and western blot analysis was performed to detect the expression of TH and renalase in the kidney. Compared with the WKY rats, SH rats in the baseline group had significantly increased blood pressure and markedly elevated TH protein expression (P<0.05), but dramatically reduced plasma renalase content and protein expression of renalase in the kidney (P<0.05). One week after surgery, the mean arterial pressure and TH protein expression in the surgery group was lowered compared with the baseline group and dramatically reduced when compared with the sham and control groups (P<0.05). In the surgery group, renalase levels were markedly increased compared with the baseline, sham and control groups (P<0.05). Six weeks after renal denervation, the mean arterial pressure and TH levels in the surgery group were significantly increased while the renalase content and expression were markedly reduced compared with those at week 1, however, there were no marked differences among the surgery, sham and control groups (P>0.05). Moreover, no pronounced differences in the above variables were found between the sham and control groups at any timepoint (P>0.05). Renal denervation can lower blood pressure, which may be attributed to the suppression of sympathetic nerves, increase in plasma renalase content and renalase expression in the kidney.

Effect of renal denervation procedure on left ventricular hypertrophy of hypertensive rats and its mechanisms.

PURPOSE: To investigate the effect of renal denervation (RDN) on the blood pressure, left ventricular hypertrophy and myocardial expression of TLR4/NF-κB in spontaneously hypertensive rats (SHR). METHODS: A total of 36 SHR were randomly assigned into control group (D0), RDN group (D) and sham group (S). 12 WKY rats of same age served as controls (WKY group). Rats in the D0 and WKY groups were sacrificed, but rats in the D and S group were sacrificed at one week and six weeks after surgery. The heart was collected and the left ventricle weighted followed by calculation of left ventricular mass index (LVMI). RESULTS: In the D0 group, the blood pressure, LVMI and protein expression of TLR4, NF-κB, TNF-α and IL-6 in the myocardium were markedly higher than that in the WKY group (p<0.05). In the D1 and D2 group, the LVMI, NE and protein expression of TLR4, NF-κB, TNF-α and IL-6 in the myocardium were significantly reduced (p<0.05). CONCLUSION: Renal denervation can significantly delay the progression of left ventricular hypertrophy in spontaneously hypertensive rats, which may be attributed to the not only the suppression of sympathetic activity and attenuation of pressure load but the improvement of myocardial immuno-inflammation.

Effect of renal denervation procedure on left ventricular hypertrophy of hypertensive rats and its mechanisms / Efeito da denervação renal na hipertrofia do ventrículo esquerdo de ratos hipertensos e seu mecanismo

PURPOSE: To investigate the effect of renal denervation (RDN) on the blood pressure, left ventricular hypertrophy and myocardial expression of TLR4/NF-κB in spontaneously hypertensive rats (SHR). METHODS: A total of 36 SHR were randomly assigned into control group (D0), RDN group (D) and sham group (S). 12 WKY rats of same age served as controls (WKY group). Rats in the D0 and WKY groups were sacrificed, but rats in the D and S group were sacrificed at one week and six weeks after surgery. The heart was collected and the left ventricle weighted followed by calculation of left ventricular mass index (LVMI). RESULTS: In the D0 group, the blood pressure, LVMI and protein expression of TLR4, NF-κB, TNF-α and IL-6 in the myocardium were markedly higher than that in the WKY group (p<0.05). In the D1 and D2 group, the LVMI, NE and protein expression of TLR4, NF-κB, TNF-α and IL-6 in the myocardium were significantly reduced (p<0.05). CONCLUSION: Renal denervation can significantly delay the progression of left ventricular hypertrophy in spontaneously hypertensive rats, which may be attributed to the not only the suppression of sympathetic activity and attenuation of pressure load but the improvement of myocardial immuno-inflammation.

OBJETIVO: Investigar o efeito da denervação renal na pressão sanguínea, na hipertrofia do ventrículo esquerdo e a expressão miocárdica de TLR4/NF-kB em ratos espontaneamente hipertensos. MÉTODOS: Trinta e seis SHR ratos foram aleatoriamente distribuídos em grupo controle, grupo denervação renal (D) e grupo sham(S). 12 WKY ratos de mesma idade serviram de controle. Os ratos controles foram sacrificados, mas os ratos com denervação renal e sham foram sacrificados uma semana e seis semanas após a cirurgia. O coração foi retirado e o ventrículo esquerdo pesado seguido pelo cálculo da massa ventricular (LVMI). RESULTADOS: No grupo DO, a pressão sanguínea, LVMI e a expressão proteica de TLR4, NF-κB, TNF-α e IL-6, no miocárdio foram marcadamente maiores do que o grupo WKY (p<0,05). Nos grupos D1 e D2, o LVMI, NE e a expressão proteica de TLR4, NF-κB, TNF-α e IL-6 no miocárdio foi significantemente reduzido (p<0,05). CONCLUSÃO: A denervação renal pode significantemente retardar a progressão da hipertrofia ventricular esquerda em ratos espontaneamente hipertensos, o que pode ser atribuído não apenas pela supressão da atividade simpática e atenuação da pressão, mas pela melhora na imunoinflamação miocárdica.

[Impact of renal denervation on expression of renalase and tyrosine hydroxylase in adult rats with spontaneous hypertension].

OBJECTIVE: To investigate the impact of renal denervation on the blood pressure, plasma renalase content and expression of renalase and tyrosine hydroxylase (TH) in the idney of spontaneous hypertensive (SH) rats and to explore the role of renal denervation in lowering the blood pressure. METHODS: SH rats were randomly assigned into a baseline group, a surgery (renal denervation) group, a sham group and a control group (n=48). WKY rats matched in age (n=12) served as a baseline control group. All rats were housed until 12 weeks old. Then, the rats in the baseline group and the WKY group were sacrificed whose blood and kidney were collected for examination. In the renal denervation group, the sham group and the control group, the blood pressure was monitored continuously. One week and 6 weeks after the renal denervation, 6 rats in each group were sacrificed whose blood and kidney were collected. ELISA was employed to measure the plasma renalase and Western blot assay done to detect the expression of TH and renalase in the kidney. RESULTS: Compared with WKY rats, blood pressure significantly increased and TH protein expression markedly elevated (P<0.05) in SH rats in the baseline group, but plasma renalase content and protein expression of renalase in the kidney dramatically reduced (P<0.05). One week after the surgery, the mean arterial pressure and TH protein expression in the surgery group were lowered compared with the baseline group and dramatically reduced compared with the sham group and the control group (P<0.05). In the surgery group, the renalase level was markedly increased compared with the baseline group, the sham group, and the control group (P<0.05). Six weeks after the renal denervation, the mean arterial pressure and TH level in the surgery group were significantly increased but the renalase content and expression markedly reduced compared with those 1 week, but there were no marked differences among the surgery group, the sham group, and the control group (P>0.05). No pronounced differences in the above variables were found between the sham group and the control group at any time point (P>0.05). CONCLUSION: Renal denervation can lower the blood pressure, which may attribute to the suppression of sympathetic nerves, increase in plasma renalase content and renalase expression in the kidney.

[Research progress with renalase and cardiovascular disease].

Renalase, a novel amine oxidase, is secreted by kidney. It regulates heart function and blood pressure by degrading catecholamines. Hormones secreted by the kidney are associated with cardiovascular disease. Renalase, as a new biomarker of heart and kidney functional correlation, can lower blood pressure, protect ischemic heart muscle, improve heart function and degrade catecholamine.

[Epidemiological status and risk factors for hypertensive disorder complicating pregnancy in a district in Changsha].

OBJECTIVE: To comprehend the epidemiological status and risk factors in hypertensive disorder complicating pregnancy (HDCP) in a district in Changsha, and to provide epidemiological evidence for early intervention of HDCP. METHODS: A total of 2 148 pregnant women in a district in Changsha from 2008 to 2009 were retrospectively analyzed and the related factors were evaluated by multivariate unconditional logistic analysis. RESULTS: The incidence of HDCP was 5.21% from 2008 to 2009. Multivariate unconditional logistic regression analysis showed that age (>35 years old)(OR=6.440,95%CI:2.987-13.884), body mass index (BMI) before pregnancy (OR=2.180,95%CI:1.619-2.936), and history of diabetes or kidney disease (OR=14.221,95%CI: 3.990-50.684) were risk factors for HDCP. CONCLUSION: Age, BMI before pregnancy, and history of diabetes or kidney disease are risk factors for HDCP, while gravidity, parity, number of abortion and multiple gestation pregnancy have no obvious correlation with HDCP.