[Risk stratification and low-density lipoprotein cholesterol goal attainment rates in patients with very high-risk or extreme high-risk atherosclerotic cardiovascular diseases regarding three guidelines].

Objective: To explore the differences of risk stratification of very high-risk or extreme high-risk atherosclerotic cardiovascular diseases (ASCVD) and the attainment rates of low-density lipoprotein cholesterol (LDL-C) management targets evaluated by three different criteria, and the causal attributions of these differences. Methods: Patients with ASCVD were consecutively enrolled from January 1 to December 31 in 2019, and were evaluated for very high-risk or extreme high-risk and LDL-C goal attainment rates with 2018 American guideline on the management of blood cholesterol (2018AG), 2019 China Cholesterol Education Program (CCEP) Expert Advice for the management of dyslipidemias (2019EA) and 2020 Chinese expert consensus on lipid management of very high-risk ASCVD patients(2020EC), respectively. The causal attributions of the differences in attainment rates were analyzed as well. Results: A total of 1 864 ASCVD patients were included in this study. According to 2018AG, 2019EA and 2020EC, the proportions of the patients with very high-risk or extreme high-risk were 59.4%, 90.7%, and 65.6%, respectively. The absolute LDL-C target attainment rates were 37.2%, 15.7%, and 13.7%, respectively, the differences between each two rates were statistically significant (all P<0.001). As to the differences in attainment rates between 2020EC and 2018AG, 61.5% were due to the different LDL-C goal attainment values and 38.5% were caused by the different risk stratifications, while for the differences between 2020EC and 2019EA attainment rates, different LDL-C goal attainment values were responsible for 13.2%, and different risk stratifications were responsible for 86.8% of the differences. Conclusions: There are significant differences in the proportions and LDL-C attainment rates among the three different criteria for very high-risk or extreme high-risk ASCVD. 2020EC showed a moderate proportion of patients with extreme high-risk, and had the lowest LDL-C attainment rate. The differences between 2020EC and 2018AG are mainly due to the LDL-C target values, and the differences between 2020EC and 2019EA are mainly caused by the risk stratifications.

Effect of fasting blood glucose on risk of new-onset hypertension in rural Chinese population: a 15-year follow-up cohort.

BACKGROUND: The purpose of this study was to examine the correlation between fasting blood glucose and new-onset hypertension and examine any synergistically effect modification with multiple risk factors. METHODS: We conducted post-hoc analyses of repeated-measures data in the original Dongzhi osteoporosis cohort study. In total, 3985 participants without hypertension aged 25-64 years were included in the current analyses. Generalized estimating equation models were used to assess the relationship between fasting blood glucose and risk of new-onset hypertension after adjusting for pertinent covariates and autocorrelations among siblings. RESULTS: 393 men (19.4%) and 398 women (20.3%) without hypertension at the baseline developed hypertension by the end of the study period. Compared to lower baseline fasting blood glucose levels (Q1-Q3: < 5.74 mmol/L; clinical cut points: < 5.6 mmol/L), higher baseline fasting blood glucose levels (Q4: ≥ 5.74 mmol/L; clinical cut points: ≥ 5.6 mmol/L and < 7.0 mmol/L) increased the risk of new-onset hypertension significantly [(OR: 1.54, 95% CI 1.19-1.98, P < 0.001); (OR: 1.38, 95% CI 1.09-1.75, P = 0.008)] in women. Additionally, a stronger significant association was found in women with elevated fasting blood glucose on risk of new-onset of hypertension with higher total cholesterol (≥ 5.2 mmol/L) [(OR: 2.76; 95% CI: (1.54, 4.96), P < 0.001)]. However, no association was found between fasting blood glucose and risk of new-onset hypertension in men. CONCLUSIONS: High fasting blood glucose may be significantly associated with risk of new-onset hypertension in Chinese women, especially in women with higher total cholesterol. Further randomized studies are needed to confirm our findings.

[Difference on prevalence and trends of conventional risk factors for ASCVD between young Chinese and American adults with first acute myocardial infarction].

Objective: This study aimed to compare the prevalence and trends of conventional risk factors for atherosclerotic cardiovascular disease(ASCVD) between young Chinese and American adults with first acute myocardial infarction. Methods: This was a retrospective cohort analysis. Hospitalized yang adults (aged from 18 to 44 years old) with first acute myocardial infarction(AMI) from January 2007 through December 2017 were identified from Beijing Anzhen hospital medical record system. Prevalence and trends of hypertension, diabetes mellitus, obesity, smoking, and dyslipidemia were analyzed and compared with young American adults, whose data were reported by Yandrapalli et al, and the hospitalizations for a first AMI in young adults aged 18 to 44 years were identified from national inpatient sample from January 2005 through September 2015. Results: Chinese cohort included 2 866 young adults with a first AMI (male, n=2 739, female, n=127), the mean age was (39±5) years. Presentation of AMI was more frequently ST-segment elevation myocardial infarction (77.3%, 2 214/2 866). American cohort included 280 875 subjects (male, n=203 700, female, n=77 175), the mean age was 39±5 years. Presentation of AMI was more frequently non-ST-segment elevation myocardial infarction (53.6%, 150 549/280 875). In China, dyslipidemia 2 254 (78.6%), smoking 2 084(72.7%), and hypertension 1 170 (40.8%) were most prevalent, and 96.0% (2 752/2 866) of patients had at least 1 risk factor; in the United States, smoking 159 537(56.8%), dyslipidemia 145 212 (51.7%), and hypertension 139 876 (49.8%) were most prevalent, and 90.3% (253 630/280 875) of patients had at least 1 risk factor. Women had a prevalence of diabetes was higher in women, and prevalence of dyslipidemia and smoking was higher in men in China (all P<0.05);prevalence of obesity, diabetes and hypertension was higher in Women, and prevalence of dyslipidemia and smoking was higher in man in the United States (all P<0.001). Prevalence of dyslipidemia and smoking was higher Chinese men (79.3% vs. 54.6%, 75.5% vs. 58.1%,all P<0.001), and prevalence of obesity, diabetes and hypertension was lower (13.1% vs. 18.6%, 14.9% vs. 19.9%, 40.6% vs. 49.3%, all P<0.001)in Chinses cohort than those in the United States cohort. Prevalence of smoking and obesity was lower (12.6% vs. 53.4%, 10.2% vs. 26.9%, all P<0.001) and prevalence of dyslipidemia was higher (63.8% vs. 44.1% P<0.001) in Chinese women than those in the United States women. Patients presenting with non-ST-segment elevation myocardial infarction had a higher prevalence of obesity, diabetes and hypertension than patients presenting with ST-segment elevation myocardial infarction in China (20.1% vs.10.9%, 17.6% vs. 14.5%, 47.4% vs. 38.9%, all P<0.05). The prevalence of the three risk factors also was higher in patients presenting with non-ST-segment elevation myocardial infarction in the US (24.0% vs.17.0%, 25.0% vs. 20.0%, 54.6% vs. 44.2%, all P<0.001), prevalence of smoking and dyslipidemia was lower in these patients (53.5% vs. 60.5%,51.4% vs. 52.1%, all P<0.001). The prevalence of hypertension and obesity increased and the rate of smoking reduced in China from 2007 through 2017 (all trend P<0.001). The prevalence of all these five conventional risk factors increased temporally in the United States from 2005 to 2015 (all trend P<0.001). The prevalence of hypertension increased by 15.6% in China and 14.5% in the United States, respectively, accounting the largest increase. Conclusions: Smoking, dyslipidemia, and hypertension are most prevalent in China and United State. Significant sex and AMI subtype difference are observed for individual risk factors. The prevalence of hypertension and obesity increased significantly over time in China and all these five conventional risk factors increased significantly in the United States.

Predictors of testicular sperm retrieval in patients with non-obstructive azoospermia: a review.

Azoospermia is divided into two categories of obstructive azoospermia and non-obstructive azoospermia. Before 1995, couples with a male partner diagnosed with non-obstructive azoospermia had to choose sperm donation or adoption to have a child. Currently, testicular sperm aspiration or micro-dissection testicular sperm extraction combined with intracytoplasmic sperm injection allows patients with non-obstructive azoospermia to have biological offspring. The sperm retrieval rate is significantly higher in micro-dissection testicular sperm extraction compared with testicular sperm aspiration. Additionally, micro-dissection testicular sperm extraction has the advantages of minimal invasion, safety, limited disruption of testicular function, a low risk of postoperative intratesticular bleeding, and low serum testosterone concentrations. Failed micro-dissection testicular sperm extraction has significant emotional and financial implications on the involved couples. Testicular sperm aspiration and micro-dissection testicular sperm extraction have the possibility of failure. Therefore, predicting the sperm retrieval rate before surgery is important. This narrative review summarizes the existing data on testicular sperm aspiration and micro-dissection testicular sperm extraction to identify the possible factor(s) that can predict the presence of sperm to guide clinical practice. The predictors of surgical sperm retrieval in patients with non-obstructive azoospermia have been widely studied, but there is no consensus.

Radiographic horizontal gaze deviation in the setting of acute PICA territory ischemia: A potential mimic of large vessel occlusion.

PURPOSE: Horizontal gaze deviation (HGD) is a predictor of acute large vessel occlusion (LVO) and helps to expedite the triage of patients to CTA and endovascular-capable sites. Patients with acute cerebellar ischemia, particularly involving the PICA territory, can also exhibit HGD. MATERIALS AND METHODS: We reviewed 2260 CTA stroke assessment cases between January 2016 and May 2020. Forty-six patients with CTA-proven acute PICA occlusions were identified and compared with 114 patients with acute LVO (ICA, M1, and M1/2). Both clinical and radiographic HGD were examined. The degree of radiographic HGD was measured for each patient. Site of ischemia was confirmed on subsequent MRI. RESULTS: Of the 46 patients with acute PICA occlusions, 20 (43.5%) patients had radiographic (+) HGD with either ipsilateral or contralateral gaze deviation, 6 of whom (13.0%) displayed clinical HGD. Of the 114 patients with LVO (control group), 72 (63.2%) patients had radiographic (+) HGD, all ipsilateral, 49 of whom (68.0%) displayed clinical HGD. The mean degree of HGD between PICA and LVO were 30.0° vs. 22.9°, respectively, p < 0.001; AUC = 0.68. CONCLUSION: Patients with acute PICA occlusion can exhibit either ipsilateral or contralateral HGD and a higher degree of HGD than LVO occlusion on NECT. In hyperacute stroke, the presence of radiographic HGD > 30° in the absence of ischemic changes in the MCA territory should prompt clinicians to closely evaluate for features of early ischemic changes in the cerebellar hemispheres that suggest acute PICA occlusion.

Hypovascular pancreas head adenocarcinoma: CT texture analysis for assessment of resection margin status and high-risk features.

OBJECTIVES: To determine if CT texture analysis features are associated with hypovascular pancreas head adenocarcinoma (PHA) postoperative margin status, nodal status, grade, lymphovascular invasion (LVI), and perineural invasion (PNI). METHODS: This Research Ethics Board-approved retrospective cohort study included 131 consecutive patients with resected PHA. Tumors were segmented on preoperative contrast-enhanced CT. Tumor diameter and texture analysis features including mean, minimum and maximum Hounsfield units, standard deviation, skewness, kurtosis, and entropy and gray-level co-occurrence matrix (GLCM) features correlation and dissimilarity were extracted. Two-sample t test and logistic regression were used to compare parameters for prediction of margin status, nodal status, grade, LVI, and PNI. Diagnostic accuracy was assessed using receiver operating characteristic curves and Youden method was used to establish cutpoints. RESULTS: Margin status was associated with GLCM correlation (p = 0.012) and dissimilarity (p = 0.003); nodal status was associated with standard deviation (p = 0.026) and entropy (p = 0.031); grade was associated with kurtosis (p = 0.031); LVI was associated with standard deviation (p = 0.047), entropy (p = 0.026), and GLCM correlation (p = 0.033) and dissimilarity (p = 0.011). No associations were found for PNI (p > 0.05). Logistic regression yielded an area under the curve of 0.70 for nodal disease, 0.70 for LVI, 0.68 for grade, and 0.65 for margin status. Optimal sensitivity/specificity was as follows: nodal disease 73%/72%, LVI 72%/65%, grade 55%/83%, and margin status 63%/66%. CONCLUSIONS: CT texture analysis features demonstrate fair diagnostic accuracy for assessment of hypovascular PHA nodal disease, LVI, grade, and postoperative margin status. Additional research is rapidly needed to identify these high-risk features with better accuracy. KEY POINTS: • CT texture analysis features are associated with pancreas head adenocarcinoma postoperative margin status which may help inform treatment decisions as a negative resection margin is required for cure. • CT texture analysis features are associated with pancreas head adenocarcinoma nodal disease, a poor prognostic feature. • Indicators of more aggressive pancreas head adenocarcinoma biology including tumor grade and LVI can be diagnosed using CT texture analysis with fair accuracy.

Degree of Conjugate Gaze Deviation on CT Predicts Proximal Vessel Occlusion and May Expedite Endovascular Therapy.

PURPOSE: Recent trials have demonstrated superior outcomes with combination IV-tPA and endovascular therapy (EVT) within 6 hours of symptom onset in patients with proximal vessel occlusion (ICA, M1, or proximal M1/M2) compared to IV-tPA alone. The current standard of diagnosis for consideration of EVT is CT angiogram (CTA). Unfortunately, not all hospitals are equipped with CTA, and the decision to transfer to tertiary centers is often based on nonenhanced CT. Ipsilateral conjugate gaze deviation (CGD) is associated with worse outcomes and larger infarcts in acute ischemic stroke. We predicted that the more proximal the occlusion, the higher the degree of CGD. MATERIALS AND METHODS: Over a period of 12 months, 182 consecutive patients with acute ischemic stroke treated at our institution were prospectively analyzed. Stroke locations were categorized based on CTA. Average degree of CGD was measured. Patient demographics, ASPECTS, collateral score, National Institutes of Health Stroke Scale, modified Rankin Scale, TICI score, length-of-stay, and mortality were collected. The median follow-up was 30 days. RESULTS: Out of ninety one of 182 patients with (+) CGD, 82 (90%) patients had ICA or middle cerebral artery (MCA) territory infarcts. The median was 25.0° in those with proximal occlusion and 13.7° in those with distal MCA occlusion (P < .001). A higher degree of CGD is positively correlated with proximity of vessel occlusion (correlation coefficient 0.2; P < .05). A cut-off greater than 20.25° (area under the curve = .76) showed a sensitivity of 64.0% and specificity 84.2%. CONCLUSIONS: Measuring degree of CGD may help in early identification of proximal vessel occlusions and expedite transfer for clot retrieval.

Magnetic Resonance Elastography to Assess Fibrosis in Kidney Allografts.

BACKGROUND AND OBJECTIVES: Fibrosis is a major cause of kidney allograft injury. Currently, the only means of assessing allograft fibrosis is by biopsy, an invasive procedure that samples <1% of the kidney. We examined whether magnetic resonance elastography, an imaging-based measure of organ stiffness, could noninvasively estimate allograft fibrosis and predict progression of allograft dysfunction. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Kidney allograft recipients >1 year post-transplant undergoing an allograft biopsy first underwent free-breathing, flow-compensated magnetic resonance elastography on a 3.0-T magnetic resonance imaging scanner. Each patient had serial eGFR measurements after the elastography scan for a follow-up period of up to 1 year. The mean stiffness value of the kidney allograft was compared with both the histopathologic Banff fibrosis score and the rate of eGFR change during the follow-up period. RESULTS: Sixteen patients who underwent magnetic resonance elastography and biopsy were studied (mean age: 54±9 years old). Whole-kidney mean stiffness ranged between 3.5 and 7.3 kPa. Whole-kidney stiffness correlated with biopsy-derived Banff fibrosis score (Spearman rho =0.67; P<0.01). Stiffness was heterogeneously distributed within each kidney, providing a possible explanation for the lack of a stronger stiffness-fibrosis correlation. We also found negative correlations between whole-kidney stiffness and both baseline eGFR (Spearman rho =-0.65; P<0.01) and eGFR change over time (Spearman rho =-0.70; P<0.01). Irrespective of the baseline eGFR, increased kidney stiffness was associated with a greater eGFR decline (regression r2=0.48; P=0.03). CONCLUSIONS: Given the limitations of allograft biopsy, our pilot study suggests the potential for magnetic resonance elastography as a novel noninvasive measure of whole-allograft fibrosis burden that may predict future changes in kidney function. Future studies exploring the utility and accuracy of magnetic resonance elastography are needed.

GLYX-13, a NMDA Receptor Glycine-Site Functional Partial Agonist, Attenuates Cerebral Ischemia Injury In Vivo and Vitro by Differential Modulations of NMDA Receptors Subunit Components at Different Post-Ischemia Stage in Mice.

Excessive activation of NMDA receptors (NMDARs) is implicated in pathological synaptic plasticity also known as post-ischemic long-term potentiation (i-LTP) which was produced by glutamate mediated excitotoxicity after stroke. In the past decades, many NMDARs inhibitors failed in clinical investigations due to severe psychotomimetic side effects. GLYX-13 is a NMDAR modulator with glycine site partial agonist properties and has potential protective effects on ischemic neuronal death. However, the underlying molecular mechanism of GLYX-13 attenuating the ischemic neuronal damage remains elusive. Our study was conducted to examine the molecular, cellular and behavioral actions of GLYX-13 in stroke, and further characterize the mechanism underlying the neuroprotective actions via modulation of the NMDAR subunit composition. In present study we found that in vitro oxygen-glucose deprivation (OGD) stroke model, GLYX-13 blocked i-LTP and restored the ratio of NR2A/NR2B subunit composition. The glycine site of NMDARs full coagonist D-serine completely blocked the effects of GLYX-13 on i-LTP. Besides, in vivo middle cerebral artery occlusion (MCAO) model, GLYX-13 decreased the cerebral infarct volume and reduced injury of hippocampus. Western analysis showed that GLYX-13 down-regulated the expression of phosphorylated NR2B (Tyr1472) and up-regulated phosphorylated NR2A (Tyr1325). Furthermore, GLYX-13 treatment along with NR2B specific antagonist (Ro256981) failed to exhibit any additional neuro-protective effects, whereas the application of NR2A antagonist (NVP-AAM007) abolished the neuroprotective effects of GLYX-13, which suggested that the protective action of GLYX-13 should be by its regulation of NMDAR subunit components. Our study provides important insights on the potential protective mechanism of GLYX-13 in ischemia and proposes the glycine site of NMDARs as a novel target for developing therapeutic strategies to store synaptic function in stroke.

Transport characteristics of nanoscale zero-valent iron carried by three different "vehicles" in porous media.

This study investigated the transport properties of nanoscale zero-valent iron (Fe(0)) (nZVI) carried by three vehicles: water, sodium dodecyl sulfate (SDS) solution, and SDS foam. Batch experiments were conducted to assess the sedimentation capability of nZVI particles in these three vehicles. Column experiments were conducted to investigate the transport properties of nZVI in porous media formed with different sizes of sand (0.25 mm to 0.5 mm, 0.5 mm to 0.9 mm, and 0.9 mm to 1.4 mm). Three main results were obtained. First, the batch experiments revealed that the stabilities of nZVI particles in SDS solution and SDS foam were improved, compared with that of nZVI particles in water. Moreover, the sedimentation of nZVI in foam was closely associated with the foam drainage volume. The nZVI content in foam was similar to that in the original foaming suspension, and the nZVI particle distribution in foam became significantly more uniform at a stirring speed of 3000 r/min. Second, the transport of nZVI was enhanced by foam compared with water and SDS solution for 0.25 mm to 0.5 mm diameter sand. For sand with diameters of 0.5 mm to 0.9 mm and 0.9 mm to 1.4 mm, the mobility of nZVI carried by SDS solution was optimal, followed by that of nZVI carried by foam and water. Thus, the mobility of nZVI in finer sand was significantly enhanced by foam, compared with that in coarse sand. In contrast, compared with the bare nZVI suspension and nZVI-laden foam, the spatial distribution of nZVI particles carried by SDS solution was significantly uniform along the column length. Third, the SDS concentration significantly influenced the migration of nZVI in porous media. The enhancement in the migration of nZVI carried by SDS solution was greater at an SDS dose of 0.25% compared with that at the other three doses (0.2%, 0.5%, and 1%) for sand with a 0.25 mm to 0.5 mm diameter. Increased SDS concentrations positively affected the transport of nZVI by foam for sand with a 0.25 mm to 0.5 mm diameter, and the SDS concentrations for enhancing the mobility of nZVI carried by SDS foam satisfied the following order: 1% > 0.5% > 0.25% > 0.2%. Thus, SDS solution and SDS foam were better vehicles than water for delivering nZVI particles to porous media for contamination remediation.

First Report of Bacterial Soft Rot of Horn Lian (Typhonium giganteum) Caused by Pectobacterium carotovorum subsp. carotovorum in Jilin Province of China.

Horn lian (Typhonium giganteum) is a perennial herb of the family Aracea and is commonly used for expelling phlegm and as an antispasmodic treatment. In August 2012, horn lian grown in Changchun, Jilin Province of China, exhibited soft rot disease with ~60% incidence and experienced great losses. Water-soaked and dark green lesions on leaves expanded along main veins. Semitransparent, water-soaked, and sunken lesions on stems expanded rapidly and caused the whole plant to collapse with a foul smell. Nine representative strains were isolated from infected leaves and stems on nutrient agar (NA) medium after 36 h incubation at 28°C (1). Colonies were round, shiny, grayish white, and convex on NA medium. All strains were gram-negative, non-fluorescent on King's B medium (KB), facultatively anaerobic, motile with three to six peritrichous flagella (observed by electron transmission microscope), positive for catalase and pectolytic activity test on potato slices, but negative for oxidase, urease, and lecithinase. Strains grew at 37°C and in yeast salts broth medium containing 5% NaCl. They also liquefied gelatin and reduced nitrate, but did not reduce sucrose. Strains were also negative for starch hydrolysis, malonate utilization, gas production from glucose and indole. Results were variable for the Voges-Proskauer test. The strains utilized sucrose, arabinose, fructose, D-galactose, D-glucose, inositol, lactose, D-mannose, D-mannitol, melibiose, rhamnose, salicin, trehalose, maltose, raffinose, glycerol, D-xylose, and cellobiose as carbon sources, but not melezitose, α-CH3-D-gluconate, sorbitol, or dulcitol. Species identity was confirmed by molecular characterization of one of the nine strains, DJL1-2. DNA GC content indicated by high performance liquid chromatography (HPLC) was 51.7%. The 16S rDNA sequence (KC07897) of DJL1-2 showed 99% identity to that of a Pectobacterium carotovorum subsp. carotovorum (Pcc) strain (CP001657) and the sequence of the 16S-23S rDNA spacer region (KJ623257) was 93% similar to that of another known strain of Pcc (CP003776). As a result, the strains were identified as Pcc (2). Pathogenicity of the nine strains was evaluated by spraying 1 ml of bacterial cell suspension (108 CFU/ml) onto healthy leaves and injecting 0.1 ml of cell suspension into stems of 3-year-old horn lian plants with a sterile pipette tip. Three seedlings were used for each strain and sterilized water served as negative controls. Pcc SMG-2 reference strain (from milk thistle) was also inoculated into horn lian leaves and stems. Inoculated plants were covered with plastic bags for 24 h in a greenhouse at 28 to 30°C. After 72 h, water-soaked lesions similar to the naturally infected plants were observed on leaves and stems inoculated by the nine isolated strains and Pcc SMG-2, while negative control plants remained symptomless. Biochemical tests and 16S rDNA sequence analysis confirmed that the re-isolated bacteria were Pcc. To our knowledge, this is the first report of Pcc causing bacterial soft rot of horn lian in Changchun, Jilin Province, China. References: (1) Z. D. Fang. Research Method of Phytopathology. China Agricultural Press, 1998. (2) N. W. Schaad, et al. Laboratory Guide for Identification of Plant Pathogenic Bacteria, 3rd ed. American Phytopathological Society, St. Paul, MN, 2001.

Alternaria brassicicola Causes a Leaf Spot on Isatis indigotica in China.

Chinese woad (Isatis indigotica) is a biennial herb in the Brassicaceae that is widely cultivated in China. Extracts from the roots and leaves have potential pharmaceutical use for treatment of flu, encephalitis, measles, hepatitis, and mumps (2). In June 2012, a leaf spot was observed on 1-year-old plants of I. indigotica in the medicinal garden of Jilin Agricultural University, Changchun, Jilin Province, China. More than 50% of the leaves and 100% of the plants in the garden were symptomatic. In the initial stage of infection, irregular to circular, dark gray spots, each surrounded by a chlorotic halo, appeared on leaves. The spots ranged from pinpoint to 5 mm in diameter. Some spots enlarged and coalesced, forming concentric rings. Black, sunken, fusiform lesions were observed on the petioles. Lesions gradually dried and exhibited a shot-hole appearance, and entire infected leaves desiccated. Small pieces of infected leaves and petioles were surface-disinfested in 75% ethanol for 60 s, rinsed thrice in sterilized distilled water, dried, and plated on potato dextrose agar. Olive-green mycelium developed after 2 days of incubation at 25°C, turned dark green, and covered the petri dish 10 days later. The periphery of each colony was gray and velvety. On potato carrot agar medium, conidia formed on branched chains. Conidiophores arose singly or in clusters, were straight or flexuous, separated, and measured 6.8 to 26.7 × 3.1 to 11.9 µm Conidia on host plant tissues were olivaceous, cylindrical or inverted clavate, and 25.8 to 65.2 × 10.9 to 18.3 µm Larger conidia were cylindrical or obclavate, and smaller conidia were oval. Transverse and longitudinal septa of conidia ranged from 3 to 10 and from 0 to 7 µm, respectively. A very small conidial beak or no beak was observed on each conidium. On the basis of these morphological characteristics,the fungus was identified as Alternaria brassicicola (3). A PCR assay with the ITS4 and ITS5 primers was used to amplify DNA extracted from each of four isolates (1). The sequence (567 bp) of isolate Sl-8 was submitted to GenBank (Accession No. KF531832), and showed 100% similarity to that of an A. brassicicola isolate (AF392985.1), confirming the species identification. Pathogenicity assays with 10 single-conidium isolates were done by spraying a conidial suspension (1 × 106 conidia/ml) of each isolate, or sterilized water for the control treatment, onto healthy leaves and petioles of five 3-month-old plants of I. indigotica. Inoculated and control plants were enclosed in plastic bags for 48 h. After 7 days, symptoms on inoculated plants were similar to those on the original diseased plants, while control plants remained symptomless. Re-isolation from inoculated plants produced mycelial colonies with morphological characteristics of A. brassicicola, fulfilling Koch's postulates. No fungus was isolated from control plants. A. napiformis and A. brassicae have been reported as causal agents of Alternaria leaf spot on I. indigotica in China (3). To our knowledge, however, this is the first report of A. brassicicola as a pathogen on I. indigotica in China. References: (1) N. L. Glass and G. C. Donaldson. Appl. Environ. Microbiol. 61:1323, 1995. (2) A. J. Li et al. Flora Reipublicae Popularis Sinicae Tomus 33, 1998. (3) T. Y. Zhang. Alternaria. Pages 99-100 in: Flora Fungorum Sinicorum, 2003.

First Report of Bacterial Soft Rot of Milk Thistle (Silybum marianum) Caused by Pectobacterium carotovorum subsp. carotovorum in Jilin Province of China.

Milk thistle (Silybum marianum) is an annual or biannual plant of the Asteraceae family that produces the hepaprotectant silymarin. In 2012, almost all milk thistle grown in the medicinal herbal garden of Jilin Agricultural University (Changchun, Jilin Province, China) exhibited symptoms of a previously undetected soft rot disease. Initial symptoms on stems appeared as tan, semitransparent, and water-soaked, then became sunken. The rotted lesions expanded rapidly and inner stem tissues were rotten with a foul smell. Eventually, the whole plant became black, then collapsed and died. Economic losses were significant as the seed crop was almost completely lost. Nine bacterial strains were isolated from tissues on nutrient agar (NA) medium after 36 h incubation at 28°C (1). Colonies of the nine strains were round, shiny, grayish white, and convex on NA medium. All strains were gram-negative, non-fluorescent, facultatively anaerobic, motile with two to four peritrichous flagella (observed by electron transmission microscope), positive for catalase and potato rot, but negative for oxidase and lecithinase. Strains grew at 37°C and in yeast salts broth medium containing 5% NaCl. They also liquefied gelatin. Strains were also negative for starch hydrolysis, malonate utilization, gas production from glucose, and indole. Results were variable for the Voges-Proskauer test and production of H2S from cysteine. The strains utilized esculin, fructose, D-galactose, D-glucose, inositol, lactose, D-mannose, D-mannitol, melibiose, rhamnose, salicin, trehalose, D-xylose, and cellobiose as carbon sources, but not melezitose, α-CH3-D-gluconate, sorbitol, or starch. Glycerol and maltose were only weakly utilized. Species identity was confirmed by molecular analysis of one of the strains, SMG-2. HPLC indicated a DNA GC content of 50.55%. The 16S rDNA sequence (KC207898) of SMG-2 showed 99% sequence identity to that of a Pectobacterium carotovorum subsp. carotovorum strain (DQ333384) and the sequence of the 16S-23S rDNA spacer region (KJ415377) was 95% similar to that of another known strain of P. carotovorum subsp. carotovorum (AF232684). Based on biochemical and physiological characteristics (2), as well as 16S rDNA gene analysis, the strains were identified as P. carotovorum subsp. carotovorum. Pathogenicity of the nine strains was evaluated by depositing a bacterial suspension (108 CFU/ml) on wounded stems (made with a disinfected razor blade) of 3-month-old milk thistle plants. Three plants were inoculated with each strain and three plants were treated with sterilized water as negative controls. Inoculated plants were covered with plastic bags for 24 h in a greenhouse at 28 to 30°C. After 48 h, the plants inoculated with bacteria showed similar symptoms as the naturally infected plants, while control plants remained symptomless. The symptoms observed on inoculated stems were rotten and sunken tissues. Bacteria were re-isolated from the inoculated plants and confirmed to be identical to the original strains based on 16S rDNA sequence analysis. To our knowledge, this is the first report of P. carotovorum subsp. carotovorum causing bacterial soft rot of milk thistle in Changchun, Jilin Province, China. References: (1) Z. D. Fang. Research Method of Phytopathology. China Agricultural Press (In Chinese), 1998. (2) N. W. Schaad et al., eds. Laboratory Guide for Identification of Plant Pathogenic Bacteria, 3rd ed. American Phytopathological Society, St. Paul, MN, 2001.

First Report of Phoma rhei as a Pathogen of Rheum rhabarbarum in China.

Rheum rhabarbarum L., rhubarb, is a perennial herb planted mainly in Hebei, Hubei, Shanxi, Heilongjiang, and Jilin provinces as well as Inner Mongolia, China. The plant grows about 1,000 meters above sea level (4), and is used widely in China to treat constipation and gout. From June to September 2012, a leaf spot was observed on R. rhabarbarum in the medicinal garden of Jilin Agricultural University, Changchun, Jilin Province, causing significant effects on the leaves of all infected plants. In the early stage of disease development, small red lesions were visible on infected leaves, which subsequently developed into irregularly shaped or circular necrotic spots, each with a light colored center, pink-red alternating concentric rings, and surrounded by a chlorotic halo. Some lesions became perforated in the center. Lesions ranged from 1 to 15 mm in diameter. Extensive spotting resulted in general browning and yellowing of entire leaves. As lesions enlarged and coalesced, some leaves died from the margin inwards. Lesions on the stem were fusiform and sunken. Small pieces of diseased leaves and stems were surface-disinfested in 75% ethanol for 60 s, rinsed twice in distilled water, dried, and plated on potato dextrose agar (PDA). A Phoma species was isolated that produced a gray or dark gray colony after 5 to 7 days. The isolate was transferred to oatmeal agar (OA) (3). Pycnidia were dark brown to black, globose to subglobose, and 121 to 354 × 100 to 262 µm. Conidia were ellipsoidal or reniform, colorless, unicellular, and 3.8 to 6.5 × 1.7 to 4.1 µm. On the basis of these characteristics, the fungus was identified as Phoma rhei (1). A PCR assay with the ITS4 and ITS5 primers was used to amplify the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) (2). The amplified product (567 bp) was sequenced and the sequence submitted to GenBank (Accession No. KF531831). The ITS sequence exhibited 99% identity to that of a P. rhei isolate in GenBank (GU237743.1), confirming the morphological identification. Pathogenicity of eight isolates on rhubarb was confirmed by spraying a spore suspension (1 × 106 spores/ml) produced on PDA on the leaves of each 6-year-old R. rhabarbarum (cv. Boyedahuang) plant. Each isolate was inoculated onto five plants, and five plants were sprayed similarly with sterilized water as a control treatment. The plants were then covered with plastic bags for 48 h, and kept in a greenhouse (20 to 30°C with a 12-h photoperiod/day). Initial symptoms on inoculated leaves were observed 3 to 4 days after inoculation, while the control plants remained healthy. Re-isolations from lesions on the inoculated leaves, using the same protocol as the original isolations, produced fungal colonies with the same morphological characteristics as the original isolates of P. rhei, but no fungi were re-isolated from the control plants. This fungus has been found on R. rhaponticum in New Zealand (1), but to our knowledge this is the first report of P. rhei on R. rhabarbarum in China. References: (1) G. H. Boerema et al. Phoma Identification Manual. Diffferentiation of Specific and Infra-Specific Taxa in Culture. CABI Publishing. Wallingford, UK, 2004. (2) D. E. L. Cooke et al. Mycol. Res. 101:667, 1997. (3) Z. D. Fang. Research Method of Phytopathology. China Agricultural Press (In Chinese), 1998. (4) A. J. Li et al. Flora Reipublicae Popularis Sinicae. Tomus 25:171, 1998.

Protective effects of IL-4 on Bacillus Calmette-Guerin and lipopolysaccharide induced immunological liver injury in mice.

OBJECTIVE: Mice injected with Bacillus Calmette-Guérin (BCG) were challenged with lipopolysaccharide (LPS) to induce inflammatory liver injury. This study was performed to explore the protective effects of interleukin (IL)-4 against liver injury induced by BCG and LPS in mice. MATERIALS AND METHODS: Mice injected with BCG (125 mg/kg) were challenged with LPS (10 µg/kg) to induce the model of inflammatory liver injury. Half an hour after injection of LPS, mice were subcutaneously administered rmIL-4 at 5 and 0.5 µg/kg, respectively. Liver injury was evaluated by serum transaminase assay and H & E staining. Liver cytokine concentrations were determined by enzyme-linked immunosorbent assay, and intrahepatic cytokine and iNOS mRNA levels by reverse transcriptase polymerase chain reaction. Intrahepatic apoptosis was evaluated by terminal deoxynucleotidyl transferase mediated nick end labeling. NF-κB p65 and ERK signal pathway was detected by Western-blotting. NF-κB signal pathway was also detected by electrophoretic mobility shift assay. RESULTS: IL-4 reduced the serum ALT, AST and LDH, alleviated the inflammatory cells infiltration, down regulated the expression of TNF-α, IL-1ß, IFN-γ, IL-6 and iNOS mRNA in liver, and alleviated hepatic glutathione depletion (GSH). In addition, IL-4 displayed inhibition of extracellular signal-regulated kinase phosphorylation and NF-κB activation. CONCLUSION: IL-4 may protect mice against BCG/LPS-induced immune liver injury, besides ERK and NF-κB signal pathways were involved in the effects.