[Value of the application of enhanced CT radiomics and machine learning in preoperative prediction of microvascular invasion in hepatocellular carcinoma].

Objective: To explore the value of machine learning models in preoperative prediction of microvascular invasion (MVI) in hepatocellular carcinoma (HCC) based on dual-phase contrast-enhanced CT radiomics features. Methods: The data of 148 patients [106 males and 42 females, with an average age of (58±11) years] with HCC confirmed by pathology in the First Affiliated Hospital of Soochow University from January 2015 to May 2020 were retrospectively analyzed, including 88 cases of positive MVI and 60 cases of negative MVI. According to the ratio of 7∶3, the patients were randomly divided into the training and validation sets, respectively. The three-dimensional (3D) radiomics features of HCC in arterial phase (AP) and portal venous phase (PP) were extracted by MaZda software, and the optimal feature subset was obtained by combining three feature selection methods (FPM method) and Lasso regression. Then, six machine learning methods were used to build the prediction models. Receiver operating characteristic (ROC) curves were drawn to evaluate the prediction ability of the aforementioned models, and the area under the curve (AUC), accuracy, sensitivity and specificity were calculated. Results: Radiomics features of HCC in AP and PP were extracted by MaZda software, with 239 in each phase. There were 7 optimal features in AP and 14 optimal features in PP selected by FPM method and Lasso regression, respectively. The AUCs of decision tree, extreme gradient boosting, random forest, support vector machine (SVM), generalized linear model, and neural network based on the 7 optimal features in AP in the validation set were 0.736, 0.910, 0.913, 0.915, 0.897, 0.648, respectively. The SVM had the highest AUC in the validation set, with the accuracy, sensitivity and specificity of 95.35%, 95.83% and 94.74%, respectively. Likewise, the AUCs of machine learning models in prediction of MVI in HCC based on the 14 optimal features in PP in the validation set were 0.873, 0.876, 0.913, 0.859, 0.877, 0.834, respectively, and there were no significant differences (all P>0.05). The random forest had the highest AUC in the validation set, with the accuracy, sensitivity and specificity of 90.70%, 87.50% and 94.74%, respectively. Conclusion: Machine learning models based on dual-phase enhanced CT radiomics features can be used in preoperative prediction of MVI in HCC, particularly the SVM and random forest models have high prediction efficiency.

Meta-analysis of the effects of furosemide combined with hydration therapy on contrast-induced acute kidney injury after coronary intervention.

OBJECTIVE: A meta-analysis was performed to evaluate the effect of furosemide combined with hydration therapy on the incidence and prognosis of contrast-induced acute kidney injury (CI-AKI) in patients after coronary intervention. MATERIALS AND METHODS: Through the PubMed, EMBASE, Cochrane Library and Web of Science databases, all relevant literature from database establishment until October 1, 2020, was retrieved and screened. Quality evaluation was performed using the risk of bias evaluation tool recommended by the Cochrane Collaboration network, data extraction was performed based on pre-selected effect indicators, and statistics were calculated using Review Manager 5.3 analysis software. RESULTS: A total of 2084 patients in 9 studies were included in the meta-analysis. The results showed that furosemide combined with hydrotherapy had no effect on the incidence of CI-AKI (OR = 0.85, 95% CI [0.46, 1.60], p = 0.62) and can significantly decrease the incidence of major adverse cardiovascular events (MACEs) (OR = 0.43, 95% CI [0.27, 0.67], p = 0.0003) and mortality (OR = 0.24, 95% CI [0.08, 0.79], p = 0.02) in patients. However, it had no significant impact on the need for postoperative dialysis treatment, postoperative creatinine level or length of hospital stay. CONCLUSIONS: Furosemide combined with hydration therapy has no significant effect on the incidence of CI-AKI in patients after coronary intervention but can reduce the incidence of MACEs and mortality, thereby providing clinical benefits.

CT-based radiomics model to distinguish necrotic hepatocellular carcinoma from pyogenic liver abscess.

AIM: To investigate the feasibility of a computed tomography (CT)-based radiomics model in distinguishing necrotic hepatocellular carcinoma (nHCC) from pyogenic liver abscess (PLA). MATERIAL AND METHODS: One hundred-four enrolled patients with nHCC (n=56) and PLA (n=48) were divided randomly into a training cohort (n=62) and validation cohort (n=42). ROI (region of interest) of the wall (ROI-wall) and ROI of the necrotic cavity (ROI-necrotic cavity) of the lesion were delineated from each arterial phase (AP) and portal venous phase (PP) image. The least absolute shrinkage and the selection operator logistic regression method was used to select radiomics features, and radiomics scores (R-scores) were calculated. Four radiomics models, including R-score (ROI-wall) in the AP, R-score (ROI-necrotic cavity) in the AP, R-score (ROI-wall) in the PP and R-score (ROI-necrotic cavity) in the PP, were constructed and evaluated by area under the curve (AUC) of receiver operating characteristic curve. RESULTS: The AUCs of R-score (ROI-wall) in the AP, R-score (ROI-necrotic cavity) in the AP, R-score (ROI-wall) in the PP, and R-score (ROI-necrotic cavity) in the PP were 0.935 and 0.917, 0.906 and 0.824, 0.985 and 0.928, 0.899 and 0.850, in the training and validation cohorts, respectively. In the training cohort, the AUC of R-score (ROI-wall) in the PP was higher than that of R-score (ROI-wall) in the AP (p=0.024) or R-score (ROI-necrotic cavity) in the AP (p=0.046) or R-score (ROI-necrotic cavity) in the PP (p=0.044). CONCLUSION: CT-based radiomics models can be used to distinguish nHCC from PLA.

[Analysis on the relationship between smoking status and the onset age of onset and the direct medical expenditure expenses of gastric cancer patients].

Objective: To investigate the relationship between smoking status and the onset age of stomach cancer patients and estimate the patients' direct medical cost burden of stomach cancer due to smoking in Anhui province. Methods: The information about the primary stomach cancer patients and their direct treatment expenditures in 10 cancer-registered areas in Anhui were collected in 2017. The association between smoking status and the age of onset of stomach cancer patients was analyzed by univariate regression and multivariate logistic regression models. The median and smoking-attributed risk method was used to describe the direct treatment expenditure of stomach cancer patients in Anhui due to smoking. Results: A total of 736 patients with stomach cancer were analyzed in this study. Univariate regression analysis showed that rural household registration (t=2.091, P=0.037), smoking (t=-2.357, P=0.001 9) and alcohol consumption (t=-2.036, P=0.042) were related to the age of onset of stomach cancer. After adjusting for gender, alcohol consumption, body mass index and household registration type, the risk of early stomach cancer in people who quitted smoking cessation was lower than that in smokers (OR=0.36, 95%CI: 0.17-0.75). The total direct medical cost burden of 736 newly diagnosed stomach cancer patients was 6.939 6 million RMB. The direct medical expenditure in stomach cancer patients who had smoking behavior was higher than that in stomach cancer patients who quitted smoking and never smoked. Conclusions: Smoking is one of the risk factors for the earlier onset of stomach cancer in Anhui. It is necessary to strengthen tobacco control to reduce the economic burden of patients with stomach cancer.

[The predictive value of gadolinium-ethoxybenzyl-diethylenetriamine pentaacetic acid enhanced MRI imaging features combined with quantitative parameters for the pathologic grading of hepatocellular carcinoma].

Objective: To investigate the predictive value of gadolinium-ethoxybenzyl- diethylenetriamine pentaacetic acid (Gd-EOB-DTPA) enhanced MRI imaging features combined with quantitative parameters for the pathologic grading of hepatocellular carcinoma (HCC). Methods: Eighty patients (65 males, 15 females; range 30-74 years,average age (59±11) years old) with HCC who underwent curative resection or biopsy from June 2016 to June 2019 in the First Affiliated Hospital of Soochow University after Gd-EOB-DTPA-enhanced MRI were evaluated retrospectively.According to the pathological results of the postoperative pathology, eighty patients were divided into poorly differentiated group (26 cases, 22 males, 4 females) and moderately-well differentiated group (54 cases, 43 males, 11 females). In Gd-EOB-DTPA-enhanced MRI, qualitative parameters were assessed. Quantitative parameters including tumor size, tumor-to-liver signal intensity ratio(SIR) of arterial phase (AP), portal vein phase (PP), equilibrium phase (EP) and hepatobiliary phase (HBP), contrast enhancement ration AP (CER-AP)were measured and calculated. Clinical data and qualitative parameters between poorly differentiated group and moderately-well differentiated group were analyzed by using χ(2) test.Quantitative parameters were analyzed by using independent sample t test. Statistically significant qualitative parameters, quantitative parameters, qualitative combined with quantitative parameters and AFP combined with qualitative and quantitative parameters were included in binary logistic regression model. The receiver operating characteristic (ROC) curves were used to evaluate the diagnostic performance of the four prediction model for the pathologic grading of HCC. Results: There was a statistical difference between poorly differentiated group and moderately-well differentiated group in AFP, arterial peritumoral enhancement, arterial rim enhancement, peritumoral hypointensity on HBP and tumor signal (all P<0.05). In two groups, tumor size, SIR-AP and SIR-PP were (7.0±3.7) vs (3.9±2.4)cm,1.11±0.29 vs 1.31±0.32 and 0.89±0.21 vs 1.03±0.27 (all P<0.05). AUC of qualitative parameters, quantitative parameters, qualitative combined with quantitative parameters and AFP combined with qualitative and quantitative parameters were 0.805, 0.804, 0.855, 0.892.There was a statistical difference between qualitative parameters and qualitative combined with quantitative parameters in sensitivity(80.8% vs 92.3%, P<0.05). Conclusion: Gd-EOB-DTPA-enhanced MRI imaging features combined with quantitative parameters can be used to predict the pathologic grading of HCC preoperatively, which has a great applicative value.

Association of thyroid cancer risk with plasma 25-hydroxyvitamin D and vitamin D binding protein: a case-control study in China.

PURPOSE: To investigate the association of thyroid cancer (TC) with 25-hydroxyvitamin D [25(OH)D] and vitamin D binding protein (DBP) and examine whether there was an interaction between 25(OH)D and DBP in relation to TC risk. METHODS: A matched case-control study based on multiple hospitals included 506 pairs of cases with newly diagnosed TC and controls. All subjects were divided into the quartiles according to the distribution of 25(OH)D and DBP in controls. Odds ratio (OR) and 95% confidence intervals (CIs) for the association of TC with 25(OH)D and DBP were estimated by conditional logistic regression. RESULTS: Comparing the highest to lowest quartiles, a decreased TC risk was respectively associated with plasma 25(OH)D (OR 0.63, 95% CI 0.40-1.00, P-trend = 0.046) and DBP (OR 0.49, 95% CI 0.32-0.77, P-trend = 0.001). However, the association between DBP and TC might be modified by 25(OH)D (P-interaction = 0.014) and physical activity (P-interaction = 0.017). Compared to participants with 25(OH)D and DBP concentrations respectively below medians, those with both concentrations above medians had a lower risk of TC (OR 0.56, 95% CI 0.39-0.82). In stratified analysis based on clinical characteristics of tumor, significantly negative association between 25(OH)D, and DBP and TC were observed among the cases with early stage of tumor progression. CONCLUSIONS: This study suggested that 25(OH)D and DBP had protective effects against TC. But the negative association between TC and DBP might be modified by 25(OH)D and physical activity.

[Superficial siderosis of the central nervous system caused by myxopapillary ependymoma of conus medullaris and cauda equine: a case report and literature review].

Superficial siderosis of the central nervous system (SSCNS) is a rare disorder caused by hemosiderin deposits in the subpial layers of the brain and spinal cord due to prolonged or recurrent low-grade bleeding into the cerebrospinal fluid (CSF). Central nervous system tumor could be one of the sources of bleeding. Some problems exist at present regarding the diagnosis and treatment of SSCNS in China. On account of fewer cases, the insufficient awareness of the condition, and the lack of long-term follow-up data, enough attention has not been paid to etiological diagnosis. The speculative high rate of missed diagnoses of SSCNS indicates a great disparity in the treatment from the world's advanced level. Related data of clinical and basic research need to accumulate as soon as possible to promote the clinical diagnosis and treatment of the disease. The progressive neurological deficits are involved in the typical clinical manifestations of SSCNS with a triad of bilateral symmetrical sensorineural hearing loss, cerebellar ataxia and signs of corticospinal tract dysfunction. Nevertheless, there are few patients with the triad signs at the same time, which lead to a delayed diagnosis or misdiagnosis. Detection of this disease was commonly post-mortem until the advent of MRI with signal and location characteristics, which made diagnosis easier. Siderosis appears as a hypointense rim covering the surface of the cerebellum, the brain stem, the spinal cord, similar to a black pencil line, thin on SE-T2-weighted images, thick and conspicuous on GE-T2-weighted images or on susceptibility-weighted imaging (SWI). The only effective way of treating the disorder is to identify the source of bleeding and remove it. MR examination is useful for seeking a source of bleeding too. Therefore, once superficial siderosis is considered, lesions of the central nervous system must be searched using MRI of the brain and spine. We report here a 37-year-old male diagnosed of SSCNS with the classical clinical symptoms of cerebellar ataxia, sensorineural hearing loss and myelopathy. T2-weighed MRI showed characteristic marginal hypo-intensity around the central nervous system. Etiological explorations revealed a large conus medullaris / cauda equina ependymoma filling the lumbosacral spinal canal, a myxopapillary ependymoma (MPE) confirmed by surgical resection and histopathological examination. The related literature was reviewed to ascertain the mechanism of SSCNS secondary to MPE, and to discuss the pathogenesis, clinical features, diagnosis and treatment of SSCNS. This paper aims to improve the awareness of SSCNS and diagnostic level, and to lay stress on the etiological explorations that is beneficial to the development of exact treatment plan.

Association of benign thyroid diseases with thyroid cancer risk: a meta-analysis of prospective observational studies.

PURPOSE: Evidence showed that benign thyroid disease was one of the risk factors for thyroid cancer. However, the results of some studies were inconsistent and were previously meta-analyses of case-control studies. Therefore, we performed a meta-analysis of prospective studies to investigate the relationship between benign thyroid diseases and thyroid cancer risk. METHODS: All eligible studies were identified via systematic searches of multiple literature databases. The combined RR (relative risk)/HR (hazard ratio) or SIR (standardized incidence ratio) with 95% confidence interval was calculated. Heterogeneity was assessed with the I2 test. Publication bias and subgroup analyses were also performed. RESULTS: Twelve studies were eligible for inclusion in the meta-analysis. The pooled RR/HR of thyroid carcinoma in benign thyroid diseases was 4.39 (95% CI 3.22-5.55). The pooled SIR of thyroid carcinoma in benign thyroid diseases was 5.98 (95% CI 4.09-7.86). Subgroup analysis was performed using the type of benign thyroid diseases. Effect value was RR/HR: hyperthyroidism (RR/HR = 3.89, 95% CI = 1.69-6.08), hypothyroidism (RR/HR = 2.72, 95% CI = 1.04-4.41), and goiter (RR/HR = 22.18, 95% CI = 12.09-32.28). Effect value was SIR: hyperthyroidism (RR/HR = 5.96, 95% CI = 1.88-10.03), goiter (RR/HR = 7.65, 95% CI = 6.94-8.37), and thyroiditis (RR/HR = 3.25, 95% CI = 1.62-4.89). CONCLUSIONS: Our study has shown that benign thyroid diseases might be associated with increased risk of thyroid cancer, especially in hyperthyroidism, hypothyroidism, and goiter. However, further investigation is needed to better understand the underlying biological mechanisms.

[Nested case-control study on risk factors of postoperative hyperactive-type delirium in geriatric orthopedic patients].

Objective: To investigate the risk factors of postoperative hyperactive-type delirium(PHTD) in geriatric orthopedic patients using nested case-control study. Methods: Seventy-four patients who underwent orthopedic surgeries in the Second Affiliated Hospital of Wenzhou Medical University from January 2008 to December 2013, aged 65 or above, American Society of Anesthesiologists (ASA) physical status Ⅰ-Ⅳ, with complete medical records and confirmed diagnosis of PHTD were selected as case group. A control group of 444 patients, on the basis of 1∶6 versus nested case group, from the same cohort but without PHTD was established. Data patterns such as patient age, gender, ASA classification, status of preoperative/postoperative electrolytes, preoperative blood glucose, preoperative/postoperative plasma albumin and preoperative/postoperative hematocrit(Hct), uses of non-steroidal analgesics, anticholinergic drugs and benzodiazepines drugs, type of anesthesia, anesthesia duration, difference in postoperative analgesia were collected and analyzed. Results: Univariate Logistic regression analysis showed that gender, age, ASA grade, preoperative electrolytes, anesthesia duration, operative duration, intraoperative hypotension and the difference of postoperative analgesia were risk factors for PHTD. Multiple Logistic regression analysis showed that gender (OR=2.562, 95%CI: 1.438-4.564, P=0.001), age (OR=3.929, 95%CI: 1.788-8.633, P=0.001), preoperative electrolytes(OR=3.714, 95%CI: 2.068-6.670, P<0.001)were the independent risk factors for PHTD. Conclusion: Male, elderly patients and abnormalities of preoperative electrolyte are the independent risk factors for PHTD in geriatric orthopedic patients.

Association between vitamin D deficiency and risk of thyroid cancer: a case-control study and a meta-analysis.

PURPOSE: Although vitamin D is reportedly associated with various cancers, the association between vitamin D and thyroid cancer is indefinite. We aimed to investigate whether this association applies to thyroid cancer (TC). METHODS: A total of 276 Chinese Han people were recruited in a current matched case-control study. Multivariable conditional logistic regression was computed to estimate the association between plasma 25(OH)D and papillary thyroid cancer (PTC). In addition, we searched relevant studies in PubMed and Web of Science databases before December 2017 to conduct a meta-analysis. RESULTS: In our case-control study, plasma 25(OH)D concentration was inversely associated with PTC risk (highest tertile vs lowest tertile: adjusted OR = 0.25; 95% CI 0.10, 0.61; Ptrend = 0.003). This association was independent of body mass index and physical activity (all adjusted Pinteraction > 0.05). A total of 11 studies were included in the meta-analysis, among which ten studies have been published and one was our case-control study. Compared with 25(OH)D non-deficient group, the pooled OR of TC was 1.42 (95% CI 1.17, 1.73) in the deficient group. Similarly, blood 25(OH)D levels in patients with TC were tend to be lower than those in the controls (SMD = - 0.20, 95% CI - 0.36, - 0.03). CONCLUSIONS: A high level of circulating 25(OH)D was associated with a decreased TC risk. This association has important significance in public health and should, therefore, be further studied.

[A concentration-response observation of hydromorphone combined with ropivacaine in labor analgesia].

Objective: To investigate the median effective dose (ED(50)) of hydromorphone and the appropriate concentration of ropivacaine combined with hydromorphone in epidural labor analgesia. Methods: One hundred and forty nulliparous women undergoing labor selected for delivery with epidural analgesia were enrolled in our hospital from January to June 2016. The first of top 50 women received 0.12% ropivacaine plus 20 µg/ml hydromorphone complex solution, then sequential women were used the modified sequential method to determine the ED(50) and ED(95) of hydromorphone. The other 90 women were randomly divided and receieved 0.08% ropivacaine and 15 µg/ml hydromorphone(H1 group), 0.10% ropivacaine and 15 µg/ml hydromorphone (H2 group), 0.12% ropivacaine and 15 µg/ml hydromorphone (group H3) respectively for epidural labor analgesia. In the course of labor, block levels of epidural analgesia, the Bromage scores, analgesia scores and fetal heart rate-uterine concraction were monitored. In addition, onset time of anesthesia, labor time, mode of delivery, cases of increased oxytocin using, neonatal Apgar score, incidence of nausea and vomiting, itching and fetal heart reduction were recorded. Results: The ED(50) and ED(95) values of hydromorphone were 10.49 (95% CI: 8.89-11.79) and 15.15 (95% CI: 13.25-22.25) µg/ml respectively. The onset time in group H1 was significantly longer than those in group H2 and H3((14.23±3.82) , ( 11.32±2.16), (10.83±2.56)min, respectively), the difference was statistically significant (t=5.854, 6.212, all P<0.05). Analgesic VAS score at 30, 60 and 90 min time points in H1 group was significantly higher than that in H2 group and H3 group (all P<0.05). VAS score at withdrawal in H1 group was significantly higher than that in group H3 ( (3.25±0.75) vs (0.27±0.12) ), the difference was statistically significant ( t=9.314, P<0.05). VAS scores at the fourth, fifth, sixth contractions after analgesia in H1 group were significantly higher than those in H2 and H3 groups (all P<0.05). The incidence of motor nerve block in group H3 was higher than that in group H1 and group H2 (26.67%, 6.66%, 3.33%, respectively), the difference was statistically significant (χ(2)=6.413, 4.320, all P<0.05). Conclusions: 0.10% ropivacaine combined with 15 µg/ml hydromorphone has a good analgesic effect, slight motor block, high safety and worthy clinical application for labor analgesia.

Fitness and Competitive Ability of Botrytis cinerea Isolates with Resistance to Multiple Chemical Classes of Fungicides.

Resistance to multiple chemical classes of fungicides in Botrytis cinerea isolates from eastern United States strawberry fields is common and strategies to control them are needed. In this study, we compared fitness and competitive ability of eight sensitive isolates (S), eight isolates resistant to five or six chemical classes of fungicides but not to phenylpyrroles (5CCR), and eight isolates resistant to six or seven chemical classes including phenylpyrroles (6CCR/MDR1h). The latter included the MDR1h phenotype due to overexpression of atrB based on Δ497V/L in mrr1. The 6CCR/MDR1h isolates grew more slowly at 4°C on potato dextrose agar, and both 5CCR and 6CCR/MDR1h isolates were hypersensitive to osmotic stress compared with S isolates. In contrast, no differences were found in oxidative sensitivity, aggressiveness, and spore production in vivo, and sclerotia production and viability in vitro. In competition experiments, the 5CCR and 6CCR/MDR1h isolates were both outcompeted by S isolates and 6CCR/MDR1h isolates were outcompeted by 5CCR isolates in the absence of fungicide pressure. Under selective pressure of a fludioxonil/pyraclostrobin rotation, the 6CCR/MDR1h isolates dominated after coinoculation with 5CCR and S isolates. The competitive disadvantage of 5CCR and especially 6CCR/MDR1h isolates suggest that, in the absence of fungicide selection pressure, S isolates may reduce inoculum potential of multifungicide-resistant isolates under field conditions.

Sensitivity of Colletotrichum Species, Including C. fioriniae and C. nymphaeae, from Peach to Demethylation Inhibitor Fungicides.

Few fungicides are effective against anthracnose, caused by Colletotrichum spp., and emerging resistance makes the search for chemical alternatives more relevant. Isolates of the Colletotrichum acutatum species complex were collected from South Carolina and Georgia peach orchards and phylogenetic analysis of the combined internal transcribed spacer region, glyceraldehyde-3-phosphate dehydrogenase, and ß-tubulin gene sequences separated the isolates into C. nymphaeae and C. fioriniae. The sensitivity of these and three other previously reported Colletotrichum spp. from peach, including C. fructicola, C. siamense, and C. truncatum, to demethylation inhibitor (DMI) fungicides difenoconazole, propiconazole, tebuconazole, metconazole, flutriafol, and fenbuconazole was determined based upon mycelial growth inhibition. C. truncatum was resistant to tebuconazole, metconazole, flutriafol, and fenbuconazole and C. nymphaeae was resistant to flutriafol and fenbuconazole based on 50% effective concentration (EC50) values >100 µg/ml. C. fructicola and C. siamense were sensitive to all DMI fungicides (EC50 values of 0.2 to 13.1 µg/ml). C. fioriniae subgroup 2 isolates were less sensitive to DMI fungicides (EC50 values of 0.5 to 16.2 µg/ml) compared with C. fioriniae subgroup 1 (EC50 values of 0.03 to 2.1 µg/ml). Difenoconazole and propiconazole provided the best control efficacy in vitro to all five species, with EC50 values of 0.2 to 2.7 µg/ml. Tebuconazole and metconazole were effective against all Colletotrichum spp., except for C. truncatum. The strong in vitro activity of some DMI fungicides against Colletotrichum spp. may be exploited for improved anthracnose disease management of peach.

First Report of Leaf and Sheath Spot Caused by Waitea circinata var. zeae on Paspalum vaginatum and Zoysia tenuifolia in China.

Waitea circinata var. zeae was previously reported as the causal agent of leaf and sheath spot on Festuca arundinacea (1) and Panicum tennesseense (2) in the United States. In late May to mid-September 2013, a disease resembling leaf and sheath spot was observed on Paspalum vaginatum in fairways from several golf courses in Hainan Province, China. Affected plants initially had large yellow and brown spots on leaves and sheathes, then the whole plant turned yellowish-brown and eventually died. The same symptoms were also observed on a lawn established with Zoysia tenuifolia in Hainan University. Symptomatic leaves were surface sterilized in 1% hypochlorite for 1 min, rinsed with sterile water three times, and plated onto potato dextrose agar (PDA) amended with 0.01% gentamicin sulfate. Two Rhizoctonia-like fungal isolates were obtained from the diseased P. vaginatum (Isolate no. ML-WC1) and Z. tenuifolia (Isolate no. HNU-1) samples. After incubation on PDA for 1 week at 25°C, white mycelial colonies developed and eventually turned a salmon color with age. Small, white, spherical sclerotia (0.5 to 1 mm in diameter) were observed submerged throughout the agar media after incubation for 1 week and turned a reddish-brown color within 4 weeks. The two isolates were tentatively identified as W. circinata var. zeae based on these characteristics. The internal transcribed spacer (ITS) region of the ribosomal DNA gene was amplified and sequenced using primer pair ITS1/ITS4. The sequences of the two isolates (GenBank Accession Nos. KJ717943 and KJ717944) were more than 99% similar to W. circinata var. zeae (JQ688056 and JQ688059) sequences deposited in GenBank. To confirm pathogenicity, inocula were prepared by incubating autoclaved rye grains with strains ML-WC1 and HNU-1 for 2 weeks at 25°C. Ten colonized rye grains were uniformly spread around the crowns of 6-week-old P. vaginatum and Z. tenuifolia seedlings grown in 10-cm-diameter pots. Each isolate was placed in four separate pots and four control plants were treated with non-inoculated grain. All pots were covered with translucent plastic bags and placed in a greenhouse at 24 ± 2°C with a 12-h light/dark cycle. By 1 week post-inoculation, significant blighting of leaves and sheaths was observed, while non-inoculated plants showed no symptoms. W. circinata var. zeae was successfully re-isolated from symptomatic plants and confirmed by its Rhizoctonia-like mycelium and small, reddish-brown, spherical sclerotia on the PDA. Recently a related species, W. circinata var. circinata, causing brown ring patch on two cool-season grasses was reported in China (3). However, the isolates of W. circinata var. zeae were distinguished from W. circinata var. circinata base on ITS sequence data and morphological characters. To our knowledge, this is the first report of W. circinata var. zeae infecting P. vaginatum and Z. tenuifolia in China. References: (1) S. S. Martin, Jr. and L. T. Lucas. Plant Dis. 67:676, 1983. (2) N. A. Mitkowski. Plant Dis. 87:1006, 2003. (3) X. X. Ni et al. Plant Dis. 96:12, 2012.

First Report of Brown Rot of Apple Caused by Monilinia fructicola in Germany.

Monilinia fructicola (G. Wint.) Honey is a causal agent of brown rot of stone fruits but may also affect pome fruits. M. fructicola is common in North America, Oceania, and South America as well as in Asia, but it is listed as a quarantine pathogen in Europe (3). Since its first discovery in Europe in 2001 (France), it has been reported in Spain, Slovenia, Italy, and Switzerland. Recently, the fungus was also detected in orchards of blackberries and plums in the State of Baden-Württemberg, Germany (4). In July 2010, apples (Malus domestica Borkh.) of the cultivar Jonagold were found in a residential backyard in Fronhausen an der Lahn located in the State of Hessen, Germany with symptoms resembling brown rot caused by Monilinia species. Affected apples were at or near maturity with brown decay that had spread throughout the fruits. On the surface of the decaying apples was tan to white zones of sporulation. Upon isolation, the mycelium grew at a linear rate of 9.2 mm per day at 22°C on potato dextrose agar forming branched, monilioid chains of grayish colonies with concentric rings and little sporulation. The lemon-shaped spores had an average size of 14 × 9 µm, a shape and size consistent with M. fructicola. The ribosomal ITS1-5.8S-ITS2 region was PCR-amplified from genomic DNA obtained from mycelium using primers ITS1 and ITS4. A BLAST search in GenBank revealed highest similarity (99%) to M. fructicola sequences from isolates collected in China, Italy, and Slovenia (GenBank Accession Nos. FJ515894.1, FJ411109.1, GU967379.1). The M. fructicola sequence from the apple isolate was submitted to GenBank (Accession No. JF325841). The pathogen was also identified to the species level and confirmed to be M. fructicola using two novel PCR techniques based on cytochrome b sequences (1,2). Pathogenicity was confirmed by inoculating three surface-sterilized, mature apples cv. Gala with a conidial suspension (105 spores/ml) of the apple isolate. Fruit were stab inoculated at three equidistant points to a depth of 10 mm using a sterile needle. A 30-µl droplet was placed on each wound; control fruit received sterile water without conidia. After 5 days of incubation at room temperature in air-tight plastic bags, the inoculated fruits developed typical brown rot symptoms with sporulating areas (as described above). The developing spores on inoculated fruit were confirmed to be M. fructicola. All control fruits remained healthy. To our knowledge, this is the first report of M. fructicola on apple in Germany and more indication of further geographical spread of the quarantine disease in Germany. References: (1) J.-M. Hily et al. Pest Manag. Sci. Online publication. doi 10.1002/ps.2074, 2011. (2) S. Miessner and G. Stammler. J. Plant Dis. Prot. 117:162, 2010. (3) OEPP/EPPO. EPPO A2 list of pests recommended for regulation as quarantine pests. Version 2009-09. Retrieved from http://www.eppo.org/QUARANTINE/listA2.htm , September 22, 2010. (4) OEPP/EPPO. Reporting Service. No. 1, January 2010. Retrieved from http://archives.eppo.org/EPPOReporting/2010/Rse-1001.pdf , September 22, 2010.

First Report of Brown Rot of Peach Caused by Monilinia fructicola in Southeastern China.

In 2009 and 2010, peaches (Prunus persica) with brown rot symptoms were collected from Zhejiang Plant Protection State Research Farm and a commercial orchard in Fujian Province in southeastern China. Affected fruit showed brown decay with zones of sporulation. Single-spore isolates from the diseased fruit were cultured on potato dextrose agar. After incubation at 25°C in the dark for 5 days, colonies were gray with concentric rings of sporulation. Mean mycelial growth of isolates MZ09-2a from Zhejiang Province and 0907-a from Fujian Province was 4.46 ± 0.58 and 7.05 ± 0.81 cm after 4 and 7 days of incubation, respectively. Lemon-shaped conidia were formed in branched, monilioid chains and mean size was 14.6 (9.6 to 21.6) × 10.3 (7.2 to 13.2) µm. Mean conidial germination was 97 ± 1% with one straight germ tube per conidium. These characteristics were consistent with descriptions of Monilinia fructicola (G. Wint.) Honey (3). Morphology-based species identification was confirmed by sequencing and analysis of ribosomal internal transcribed spacer (ITS) sequences. A 496-bp fragment including ITS 1 and 2 and the gene encoding the 5.8S small subunit of the ribosomal RNA from isolates MZ09-2a and 0907-a was amplified using the universal primer pair ITS1/ITS4 (4) and sequenced. Nucleotide sequences of both isolates were identical. Blast searches of the ITS sequences in GenBank showed the highest similarity (100%) with sequences of M. fructicola isolates from China (FJ515894), Italy (FJ411109), and Spain (EF207423). The isolates were also identified as M. fructicola using the Monilinia spp. PCR detection protocol based on sequence-characterized amplification region marker DNA sequences (2). Pathogenicity was confirmed by inoculating surface-sterilized, mature cv. Zhonghua 2 peaches with mycelial plugs of representative isolates. Fruit was stabbed at two points with a 5-mm-diameter sterile cork borer, mycelial plugs (5 mm in diameter) were removed from the periphery of a 4-day-old colony of each isolate and placed upside down into each wound; control fruit received water agar. Inoculated fruit developed typical brown rot symptoms with sporulating fungi while control fruit remained healthy after 3 days of incubation at 22°C in a moist chamber. Pathogens were reisolated from the inoculated fruit and confirmed to be M. fructicola on the basis of morphological characteristics. To our knowledge, this is the first report of M. fructicola in Zhejiang and Fujian provinces. Both provinces are located more than 1,000 km south of Beijing, Hebei, and Shandong provinces, where M. fructicola had been reported previously (1). References: (1) J. Y. Fan et al. Acta Phytophylacica Sin. (in Chinese) 34:289, 2007. (2) I. Gell et al. J. Appl. Microbiol. 103:2629, 2007. (3) G. C. M. van Leeuwen and H. A. van Kesteren. Can. J. Bot. 76:2041, 1998. (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, 1990.