[Inhibition connexin 43 by mimetic peptide Gap27 mediates protective effects on 6-hydroxydopamine induced Parkinson's disease mouse model].

OBJECTIVE: To explore whether the using of mimetic peptide Gap27, a selective inhibitor of connexin 43 (Cx43), could block the death of dopamine neurons and influence the expression of Cx43 in 6-hydroxydopamine (6-OHDA)-induced Parkinson's disease mouse models. METHODS: Eighteen C57BL/6 mice were randomly divided into control group, 6-OHDA group and 6-OHDA+Gap27 group, with 6 mice in each group. Bilateral substantia nigra stereotactic injection was performed. The control group was injected with ascorbate solution, 6-OHDA group was injected with 6-OHDA solution, and 6-OHDA+Gap27 group was injected with 6-OHDA and Gap27 mixed solution. Immuno-histochemical staining was used to detect the number of dopamine neurons, quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of Cx43 messenger ribonucleic acid (mRNA), immuno-fluorescence staining was used to detect the distribution of Cx43 protein, the contents of Cx43 protein and Cx43 phosphorylation at serine 368 (Cx43-ps368) in mouse midbrain were detected by Western blot. RESULTS: After injection of 6-OHDA, numerous dopamine neurons in substantia nigra died as Cx43 content increased, Cx43-ps368 content decreased. Mixing Gap27 while injecting 6-OHDA could reduce the number of death dopamine neurons and weaken the changes of Cx43 and Cx43-ps368 content caused by 6-OHDA. The number of tyrosine hydroxylase (TH) immunoreactive positive neurons in 6-OHDA group decreased to 27.7% ± 0.02% of the control group (P < 0.01); The number of TH immunoreactive positive neurons in 6-OHDA+Gap27 group was (1.64±0.16) times higher than that in 6-OHDA group (P < 0.05); The content of total Cx43 protein in 6-OHDA group was (1.44±0.07) times higher than that in 6-OHDA+Gap27 group (P < 0.05) while (1.68±0.07) times higher than that in control group (P < 0.01). In 6-OHDA group, the content of Cx43-ps368 protein and its proportion in total Cx43 protein were significantly lower than that in 6-OHDA+Gap27 group (P < 0.05). CONCLUSION: In 6-OHDA mouse models, mimetic peptide Gap27 played a protective role in reducing the damage to substantia nigra dopamine neurons, which was induced by 6-OHDA. The overexpression of Cx43 protein might have neurotoxicity to dopamine neuron. Meanwhile, decreasing Cx43 protein level and keeping Cx43-ps368 protein level may be the protective mechanisms of Gap27.

[Effect of benzo(a)pyrene on dopaminergic neurons and α-synuclein in brain and its mechanism involved].

OBJECTIVE: To analyze the effect of benzopyrene on the decrease of dopaminergic neurons, and the increase and aggregation of α-synuclein, which are the pathological features of Parkinson's disease, and to explore its possible mechanisms. METHODS: Eight-month-old transgenic mice with human SNCA gene were randomly divided into a BaP-exposed group and a control group. BaP and solvent corn oil were injected intraperitoneally to BaP-exposed group and control group respectively, once a day for 60 days. The motor dysfunction of mice was tested by rotarod test. The effects of BaP on the decrease of dopaminergic neurons and increase and aggregation of α-synuclein were observed by immunohistochemistry and Western blot experiments respectively, and the expression of related mRNA was detected by quantitative real-time PCR (qRT-PCR). Twenty genes were tested in the study, mainly related to neurotransmitter transporter (2 genes), neurotransmitter receptor function (10 genes), cellular autophagy (5 genes), and α-synuclein aggregation and degradation (3 genes). RESULTS: After BaP exposure, the movement time of the mice in the rotarod test was significantly reduced (P<0.05). The substantia nigra dopami-nergic neurons in the mice were significantly reduced, which was 62% of the control group (P<0.05), and the expression of α-synuclein in the midbrain increased, which was 1.36 times that of the control group (P<0.05). After BaP exposure, mRNA expressions of 14 genes in the midbrain of the mice were significantly down-regulated (P<0.05). Alpha-synuclein degradation and cell autophagy (5 genes), neuron transporters (2 genes), and neurotransmitter receptor functions (5 genes) were involved. The expression of one gene, Synphilin-1, was significantly up-regulated (P<0.01), which was related to α-synuclein aggregation. CONCLUSION: BaP exposure not only inhibited function of neurotransmitter receptor and dopamine transporter, but also interfered cell autophagy, thereby hindering the degradation of α-synuclein, which could lead to decrease of dopaminergic neurons in substantia nigra and increase and aggregation of α-synuclein in midbrain, as the significant pathology of Parkinson's disease. Therefore, BaP exposure may increase the risk of Parkinson's disease.

High diversity of Diaporthe species associated with pear shoot canker in China.

Species of Diaporthe (syn. Phomopsis) are important endophytes, saprobes and pathogens, infecting a wide range of plants and resulting in important crop diseases. However, the species occurring on pear remain largely unresolved. In this study, a total of 453 Diaporthe isolates were obtained from branches of Pyrus plants (including P. bretschneideri, P. communis, P. pyrifolia and P. ussuriensis collected from 12 provinces in China) showing shoot canker symptoms. Phylogenetic analyses based on five loci (ITS, TEF, CAL, HIS, and TUB) coupled with morphology of 113 representative isolates revealed that 19 Diaporthe species were isolated, representing 13 known species (including D. caryae, D. cercidis, D. citrichinensis, D. eres, D. fusicola, D. ganjae, D. hongkongensis, D. padina, D. pescicola, D. sojae, D. taoicola, D. unshiuensis and D. velutina) and six new species described here as D. acuta, D. chongqingensis, D. fulvicolor, D. parvae, D. spinosa and D. zaobaisu. Although Koch's postulates confirmed all species to be pathogenic, a high degree of variation in aggressiveness was observed. Moreover, these species have a high diversity, plasticity, and prevalence related to the geographical location and pear species involved.

Risk factors and score for recollapse of the augmented vertebrae after percutaneous vertebroplasty in osteoporotic vertebral compression fractures.

Our study demonstrated a high incidence of recollapse of the augmented vertebrae after PVP treatment for OVCFs. A risk score based on all significant factors can predict the rate of recollapse and gain clinical benefits to prevent recollapse in patients at high risk. BACKGROUND: Recollapse of the augmented vertebrae after percutaneous vertebroplasty (PVP) treatment for osteoporotic vertebral compression fractures (OVCFs) has obtained much attention. However, little is known about risk factors and score for recollapse of the augmented vertebrae. OBJECTIVE: To determine risk factors and furthermore develop a risk score related to recollapse of the augmented vertebrae after PVP treatment for OVCFs. METHODS: Patients who were treated with PVP for single OVCFs and met this study's inclusion criteria were retrospectively reviewed. The follow-up period was at least 2 years. Associations of recollapse with co-variates (age, gender, bone mass density [BMD] with a T-score, fracture level, intravertebral cleft [IVC], fracture type, cement volume, cement leakage, leakage into a disc, cement distribution pattern, Non-PMMA-endplate-contact [NPEC], preoperative fracture severity, reduction rate [RR], reduction angle [RA]) were analyzed and a risk score for recollapse was further developed to predict recollapse. RESULTS: A total of 152 patients were included. Recollapse group was found in 42 (27.6%) patients. Preoperative IVC, solid lump cement distribution pattern, more RR (a cutoff value of 7%) and larger RA (a cutoff value of 3°) was significantly associated with increased risk for recollapse of the augmented vertebrae. A risk score was developed based on the number of risk factors present in each patient. Patients with a score of 4 had an approximately ninefold increased risk of developing recollapse over patients with a score of 0. The receiver operating characteristic curve of the risk score generated an area under the curve of 0.899 (95% CI 0.642-0.836, P = 0.000). CONCLUSION: A risk score based on preoperative IVC, cement distribution pattern, reduction rate, and reduction angle predicts the rate of recollapse. Additional studies should aim to validate this score and inspect clinical benefits of recollapse prophylaxis in patients at high risk.

Colletotrichum species associated with anthracnose of Pyrus spp. in China.

Colletotrichum species are plant pathogens, saprobes, and endophytes on a range of economically important hosts. However, the species occurring on pear remain largely unresolved. To determine the morphology, phylogeny and biology of Colletotrichum species associated with Pyrus plants, a total of 295 samples were collected from cultivated pear species (including P. pyrifolia, P. bretschneideri, and P. communis) from seven major pear-cultivation provinces in China. The pear leaves and fruits affected by anthracnose were sampled and subjected to fungus isolation, resulting in a total of 488 Colletotrichum isolates. Phylogenetic analyses based on six loci (ACT, TUB2, CAL, CHS-1, GAPDH, and ITS) coupled with morphology of 90 representative isolates revealed that they belong to 10 known Colletotrichum species, including C. aenigma, C. citricola, C. conoides, C. fioriniae, C. fructicola, C. gloeosporioides, C. karstii, C. plurivorum, C. siamense, C. wuxiense, and two novel species, described here as C. jinshuiense and C. pyrifoliae. Of these, C. fructicola was the most dominant, occurring on P. pyrifolia and P. bretschneideri in all surveyed provinces except in Shandong, where C. siamense was dominant. In contrast, only C. siamense and C. fioriniae were isolated from P. communis, with the former being dominant. In order to prove Koch's postulates, pathogenicity tests on pear leaves and fruits revealed a broad diversity in pathogenicity and aggressiveness among the species and isolates, of which C. citricola, C. jinshuiense, C. pyrifoliae, and C. conoides appeared to be organ-specific on either leaves or fruits. This study also represents the first reports of C. citricola, C. conoides, C. karstii, C. plurivorum, C. siamense, and C. wuxiense causing anthracnose on pear.

Characterization of Causal Agents of a Novel Disease Inducing Brown-Black Spots on Tender Tea Leaves in China.

A novel disease characterized by small brown-black spots (1 to 2 mm in diameter) on tender tea leaves (Camellia sinensis) has been observed in many regions of Hubei Province, China, which severely affects the yield and quality of tea. Tea leaf samples with typical symptoms were collected from three major tea-cultivation regions of Hubei, and were subjected to pathogen isolation for etiological analysis. As a result, 34 Pestalotiopsis isolates were obtained from 20 samples, and they were identified as Pestalotiopsis theae (14 isolates), P. camelliae (12), and P. clavispora (8), determined by morphologies and phylogenetic analysis based on internal transcribed spacer, and partial ß-tubulin and translation elongation factor 1-alpha genes. Pathogenicity tests on detached tea leaves showed that no matter what mycelial discs or conidium suspensions were used, inoculation of the Pestalotiopsis fungi could result in small brown-black spots (1 to 2 mm in diameter) on wounded leaves, similar to those observed in the field in the sizes and colors. It also revealed that only P. theae had pathogenicity on unwounded tea leaves, and P. theae and P. clavispora showed significantly higher virulence than P. camelliae. Inoculation test with conidium suspension on intact tea leaves in the field further confirmed that P. theae as the pathogen of brown-black spots. Reisolation of the pathogens from diseased leaves confirmed that the symptom was caused by the inoculation of Pestalotiopsis fungi. The P. theae isolates responsible for brown-black spots were also compared with those for tea gray blight disease in growth rate, pathogenicity, and molecular characteristics in parallel. To our knowledge, this is the first report that the Pestalotiopsis fungi cause brown-black spot disease on tender tea leaves. The results provide important implications for the prevention and management of this economically important disease.

[Interaction between APOB gene polymorphism and risk factors in coronary heart disease patients without lipid-lowering treatment].

Objective: To investigate the association between APOB gene R532W polymorphism and the risk of coronary heart disease (CHD) in patients without lipid-lowering treatment and to analyze the interactions between the variation of R532W and different risk factors of CHD. Methods: CHD and non-CHD were diagnosed according to coronary artery angiography (CAG) and/or coronary computed tomography angiogram (CTA) results, as well as clinical features. Blood samples from 771 CHD patients and 772 age- and sex-matched non-CHD controls, who never accepted any lipid-lowering treatments, were collected. R532W was genotyped by HumanExome BeadChip at BGI and strict quality control was made. Firstly, the association between R532W polymorphism and the risk of CHD in 3 genetic models (GA+ AA vs.GG, AA vs. GG+ GA, AA vs. GA vs. GG) after adjusting confounding factors was explored. Then, the interactions between the variation of this loci and risk factors related to CHD were investigated. Results: (1) Total cholesterol (TC) levels were significantly lower in AA genotype than in GA genotype in the total cohort and non-CHD controls, but was similar among the 3 genotypes in CHD patients. (2) R532W GG, GA and AA distribution was 80.7%, 18.2% and 1.2% in CHD patients, and 74.6%, 23.8% and 1.6% in non-CHD controls (P<0.05). (3) R532 polymorphism was related to the incidence of CHD in the dominant model, and A-allele carriers were related to about 35% reduced risk of CHD (OR=0.653, 95% CI 0.502-0.849, P=0.001) after adjusting for confounding factors. (4) R532W polymorphism had positive interactions with hypertension (1.452) and smoke (1.077), while negative interaction with diabetes (0.553) in the occurrence of CHD. Conclusions: APOB gene R532W polymorphism is related to TC levels in Chinese north Han population. A-allele carries of R532W loci is linked with reduced risk of CHD in the absence of lipid-lowering treatment. R532W polymorphism has a positive additive interaction with hypertension and smoke, while a negative additive interaction with diabetes mellitus in the occurrence of CHD.

Development and characterization of microsatellite markers for Ulmus chenmoui (Ulmaceae), an endangered tree endemic to eastern China.

Ulmus chenmoui (Ulmaceae) is an endangered tree found on Langya Mountain, eastern China. To better understand the population genetics of U. chenmoui and conserve the species, we developed microsatellite markers. Using a suppression-polymerase chain reaction technique, 74 compound microsatellite primer pairs were designed. Twelve microsatellite markers were polymorphic in 39 individuals, and the number of alleles per locus ranged from 3 to 9. The observed and expected heterozygosities ranged from 0.051 to 0.769 and from 0.533 to 0.768, respectively. Significant linkage disequilibrium was detected for three pairs of loci (P < 0.01), which may be due to a recent population bottleneck and the small population size. Nine of the 12 loci deviated from the Hardy-Weinberg equilibrium (P < 0.01), which could be explained by significant inbreeding rather than the presence of null alleles. These markers will provide a solid basis for future efforts in population genetic studies of U. chenmoui, which in turn will contribute to species conservation.

[The impact of ambient fine particulate matter on the blood pressure of an urban population in Shanghai, China: a panel study].

OBJECTIVE: To investigate the impact of particulate matter 2.5 (PM2.5) on the blood pressure of urban residents in Shanghai, China. METHODS: A panel study was conducted from May 27(th) to June 5(th) 2014 in a cohort of 30 adults in an urban community. Participants were 50-80 years old, had lived in the community for at least 5 years, and had a good health status. Key exclusion criteria were current smoking, smoking during the last 3 years, passive smoking, alcohol consumption, and severe cardiopulmonary disease. A total of 28 participants were eligible. Information on demographic characteristics, including of age, sex, height, weight, education, income and chronic comorbidities were collected. Participants were requested to have six weekly blood pressure measurements. Real-time concentrations of PM2.5 and gaseous pollutants were obtained from a nearby air quality monitor during 40 d. Pearson correlation was applied to analyze the correlation between PM2.5 and SO2, NO2, CO and O3. Linear mixed models were applied to analyze the association between PM2.5 and blood pressure, after controlling for time-related trends, day of the week, mean temperature, relative humidity and individual characteristics. RESULTS: The mean systolic and diastolic blood pressure was (124.0±15.0) mmHg (1 mmHg=0.133 kPa) and (74.0±7.7) mmHg, respectively. At 24 h before blood pressure measurement, the mean PM2.5, SO2, NO2, O3 and CO concentration were (8.3±4.9), (46.6±12.9) , (79.2±27.4) µg/m(3) and (0.8±0.2) mg/m(3). The Pearson correlation coefficient R, between PM2.5 and O3, SO2, NO2 and CO was 0.79, 0.59, 0.34 and 0.45, respectively, with corresponding P-values of <0.001, 0.006, 0.012 and 0.009, respectively. The significant correlation between PM2.5 and systolic blood pressure occurred at lag 1 day, was strongest at lag 3 day, but attenuated thereafter. A 10 µg/m(3) increase in 3-day average concentrations of PM2.5 was associated with changes of 1.86 (95%CI: 0.62-3.09) mmHg in systolic blood pressure and -0.05 (95%CI: 0.59-0.50) mmHg in diastolic blood pressure. CONCLUSION: Short-term exposure to PM2.5 may significantly elevate the systolic blood pressure of urban residents in Shanghai.

Field evidence for earlier leaf-out dates in alpine grassland on the eastern Tibetan Plateau from 1990 to 2006.

Worldwide, many plant species are experiencing an earlier onset of spring phenophases due to climate warming. Rapid recent temperature increases on the Tibetan Plateau (TP) have triggered changes in the spring phenology of the local vegetation. However, remote sensing studies of the land surface phenology have reached conflicting interpretations about green-up patterns observed on the TP since the mid-1990s. We investigated this issue using field phenological observations from 1990 to 2006, for 11 dominant plants on the TP at the levels of species, families (Gramineae-grasses and Cyperaceae-sedges) and vegetation communities (alpine meadow and alpine steppe). We found a significant trend of earlier leaf-out dates for one species (Koeleria cristata). The leaf-out dates of both Gramineae and Cyperaceae had advanced (the latter significantly, starting an average of 9 days later per year than the former), but the correlation between them was significant. The leaf-out dates of both vegetation communities also advanced, but the pattern was only significant in the alpine meadow. This study provides the first field evidence of advancement in spring leaf phenology on the TP and suggests that the phenology of the alpine steppe can differ from that of the alpine meadow. These findings will be useful for understanding ecosystem responses to climate change and for grassland management on the TP.

Functional polymorphisms in microRNAs and susceptibility to liver cancer: a meta-analysis and meta-regression.

MicroRNAs (miRNAs) are small non-coding RNA molecules that play a fundamental role in controlling a variety of biological functions. Emerging evidence has shown that common genetic polymorphisms in miRNAs may be associated with the development of liver cancer; however, several individually published studies showed inconclusive results. This meta-analysis aimed to derive a more precise estimation of the association between functional polymorphisms in miRNAs and susceptibility to liver cancer. A literature search of PubMed, Embase, Web of Science, and China BioMedicine (CBM) databases was conducted on articles published before May 1, 2012. Crude odds ratios with 95% confidence intervals were calculated. Fourteen case-control studies were included with a total of 6824 liver cancer patients and 7674 healthy controls. Nine single nucleotide polymorphisms in miRNAs were assessed, including miR-146a G>C (rs2910164), miR-499 T>C (rs3746444), miR-218 A>G (rs11134527), miR-let-7c Ins/Del (rs6147150), miR-106b-25 A>G (rs999885), miR-34b/c T>C (rs4938723), miR-196a-2 C>T (rs11614913), miR-920 Ins/Del (rs16405), and miR-122 Ins/Del (rs3783553). The meta-analysis results showed that miR-let-7c Del, miR-34b/c C, and miR-122 Del variants may be associated with increased liver cancer risk. Conversely, miR-920*Del variant may decrease the risk of liver cancer. However, miR-146a G>C, miR-196a-2 C>T, miR-499 T>C, and miR-218 A>G polymorphisms showed no significant association with liver cancer risk. In conclusion, the current meta-analysis suggests that miR-let- 7c Del, miR-34b/c C and miR-122 Del variants may be associated with increased liver cancer risk, while miR-920 Del variant may be a protective factor against liver cancer.

First Report of Alternaria alternata Causing Leaf Spots of Tea (Camellia sinensis) in China.

Tea is the most popular non-alcoholic beverage crop in the world, which originated in China and has been cultivated in over 45 countries. In recent years, a leaf spot disease of unknown etiology has been observed on young leaves of tea trees (Camellia sinensis) grown in Luotian county, Hubei Province, China. Observed symptoms display grayish brown to white spots (about 1 cm in diameter) surrounded by brown edges. Over 20% of the young leaves were affected on surveyed trees. To identify the pathogen, six symptomatic tea leaves were collected from six individual tea trees of unknown variety in August 2012. A thin section (3 to 5 mm) of symptomatic tissue was sterilized in a bleach solution of 3% hypochlorite and placed on potato dextrose agar (PDA) medium at 25°C in darkness for isolation. Six fungal colonies displaying gray-brown and gray-white aerial mycelia were consistently recovered from lesions of the six leaves, termed as T1 to T6, respectively. Conidia produced on the colonies were olive brown, obpyriform, short conical beak at the tip, 0 to 3 vertical and 1 to 6 transverse septa, and length × width of 7.1 to 31.7 (avg. 20.1) × 2.9 to 12.7 (avg. 7.2) µm. T1 to T6 were identified as Alternaria alternata on the basis of morphological characterization, respectively (2). Confirmation of the species identification was obtained by molecular characterization of their internal transcribed spacer (ITS) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) regions amplified from the genomic DNAs using the universal primers (1). The results revealed identical sequences of ITS (GenBank Accession No. KF699530) and GAPDH among the six isolates. BLAST searches showed that they had the highest similarity with A. alternata strains, with 98.3% for ITS (AJ276055) and 96.2% for GAPDH (EF513205), deposited in fungus database ( http://www.mycobank.org/ ). Pathogenicity tests were conducted on the detached leaves expanding for 10 to 20 days of two tea varieties (cvs. Fudingdabai and Taicha No. 12) in triplicate by placing 4 mm diameter discs from 5-day-old PDA plates of T3 and T6, which were incubated in an incubator at 25°C with a 12-h photoperiod for 7 days. All inoculated leaves with or without wound treatment developed brown spots similar to the original ones at 7 days post inoculation (dpi) while the control leaves inoculated with non-colonized PDA plugs remained asymptomatic. Isolates recovered from diseased samples were of the same morphology and ITS sequence as the inoculated ones. Alternaria alternata had been described on C. sinensis in China (3), but it was only reported as a severe foliar fungal pathogen of tea in North Bengal, India (1), and to our knowledge, this is the first report of A. alternata causing leaf spots on tea leaves (C. sinensis) in China. In addition to quantity loss, the species may result in a decrease of quality of tea crop considering that it can produce Alternaria toxins related to animal and public health. The etiologic identification of the disease is expected to provide useful information for its control. References: (1) B. N. Chakraborty et al. Plant Pathol. 55:303, 2006. (2) E. G. Simmons. Page 1 in: Alternaria Biology, Plant Diseases and Metabolites. J. Chelchowski and A. Visconti, eds. Elsevier, Amsterdam, 1992. (3) F. L. Tai. Page 1527 in: Sylloge Fungorum Sinicorum. eds. Sci. Press Acad. Sin. Beijing, 1979. (4) B. S. Weir et al. Stud. Mycol. 73:115, 2012.

First Report of Valsa leucostoma Causing Valsa Canker of Pyrus communis (cv. Duchess de' Angouleme) in China.

Pear is a popular fruit in the world market, and has been widely cultivated in China. Since 2008, a severe canker disease has consistently been observed on 20-year-old pear trees (Pyrus communis cv. Duchess de' Angouleme) grown in a nursery in Xingcheng, Liaoning Province, China. Observed symptoms include brown elongated ulcerative lesions (more than 20 cm in length in general), with red brown conidia produced on wet lesions. Reductions in tree vigor and yield were observed for infected trees. Tree mortality was observed for severe infections. To diagnose the pathogen, 15 canker samples were collected from five pear trees in April, 2012. Bark pieces (3 to 5 mm) taken from the border of healthy and diseased tissue were surface-disinfected with 0.1% mercury bichloride and 75% ethanol for 45 s, and placed on potato dextrose agar (PDA) medium at 25°C in darkness. Fungal colonies with a common colony morphology were consistently recovered from three samples. These fungal colonies were initially white, becoming olive green in 3 days. Conidia produced on colonies were hyaline, allantoid, and single-celled with average length × width of 6.04 (5.43 to 6.59) × 0.65 (0.51 to 0.73) µm, which were consistent with descriptions of Valsa leucostoma (1). Genomic DNA was extracted from a representative isolate F-LN-32b, and subjected to PCR amplification of the internal transcribed spacer region (ITS), ß-tubulin gene, and EF1 gene using the primer pairs ITS1/ITS4, Bt2a/Bt2b and EF1-728F/EF1-986R (3), respectively. Sequence alignment of the amplified fragments with the deposited data in NCBI showed that sequences of EF1, ITS, and ß-tubulin (GenBank Accession No. KF293296 to KF293298, respectively) of isolate F-LN-32b had the highest similarity of 99% to those of V. leucostoma strain 32-2w (JQ900340, JN584644, and JQ900374), and suggested that isolate F-LN-32b is a V. leucostoma strain. Pathogenicity tests was carried out by placing a 5-mm-diameter, 2-day-old mycelium agar plug of isolate F-LN-32b onto a punched bark hole of a detached 1-year-old pear shoot after it was surface disinfested with ethanol. Inoculated shoots were incubated at 25°C in plastic containers covered with plastic film. Pathogenicity assays were conducted on 18 pear varieties (cvs. Qiuyue, Jinshui 2, Hohsui, Huali 1, Cuiguan, Shinseiki, Xuehua, Dangshansu, Zaosu, Hongxiangsu, Yuluxiang, Nanguoli, Xizilv, Bartlett, Huanghua, Huashan, Duchess de' Angouleme, and Packham's) collected from a nursery in Wuhan, Hubei Province, China. Six shoots were inoculated for each variety and the assay was conducted three times. All inoculated shoots developed the typical canker symptoms after 6 days post inoculation (dpi) and sporulated at 25 dpi while the control shoots inoculated with non-colonized PDA plugs remained asymptomatic. Isolates recovered from inoculated samples were of the same morphology and ITS sequence as F-LN-32b. Based on these results, V. leucostoma was determined as the pathogen responsible for the Valsa canker disease on pear. Valsa mali var. pyri was identified as the only pathogen causing Valsa canker disease on pear in China (2). To our knowledge, this is the first report of V. leucostoma causing a canker disease on pear in China. References: (1) G. C. Adams et al. Australas. Plant Pathol. 35:521, 2006. (2) X. L. Wang et al. Mycologia 103:317, 2011. (3) T. J. White et al. Pages 315-322 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

[Effects of myocardial performance index on assessing left ventricular function in patients with primary hypertension].

OBJECTIVE: To investigate the value of myocardial performance index (MPI) in assessing LV function in patients with primary hypertension (HP). METHODS: We studied 130 patients with HP (mean age 54.9±13.3 years)and 155 healthy control subjects (mean age 52.4±11.6 years). MPI was determined by tissue doppler imaging using the following formula: MPI=(isovolumic contraction time + isovolumic relaxation time)/ ejection time. The HP group was divided into hypertrophy subgroup( LVMI≥115 g/m(2) in males, or ≥95 g/m(2) in females) and normal mass subgroup(LVMI <115 g/m(2) in males, or<95 g/m(2) in females). RESULTS: MPI was significantly different in control group, normal mass subgroup and hypertrophy subgroup(0.72±0.23 vs. 0.54± 0.17 vs. 0.45±0.11, P<0.001). Hypertrophy subgroup had significant higher MPI than normal mass subgroup(P =0.046), and both the groups had significant higher MPI than control group(all P<0.001). MPI was positively associated with age(r=0.369,P<0.001), Left ventricular end diastolic diameter(r=0.169, P<0.05), Sm(r=-0.211, P<0.001) and Em(r=-0.383, P<0.001) in control group. In multiple linear regression analysis, MPI was independently related to age (ß=0.492, t=7.222,P<0.001) in control group. Among the HP patients, MPI was positively associated with left atrial area (r=0.293, P<0.001),intra ventricular septum(IVS) diameter (r=0.453, P<0.001), LVMI (r=0.453, P<0.001), relative wall thickness(r=0.458, P<0.001), and negatively associated with Sm(r=-0.414, P<0.001), Em(r=-0.508, P<0.001), left ventricular ejection fraction (r=-0.305, P<0.001) in bivariate analysis. In the multiple linear regression analysis, MPI was independently related to Em (ß=0.401, t=4.256,P<0.001) and IVS diameter (ß=-0.365, t=-3.878,P<0.001) in the HP patients. CONCLUSION: The HP patients had elevated MPI, especially in the ones with LV hypertrophy. Tissue doppler imaging (TDI) derived MPI could be a useful index to evaluate the overall cardiac function in HP patients.

Low serum adiponectin levels are associated with an increased risk of diabetes in obese dogs.

OBJECTIVES: Adiponectin plays an important role in carbohydrate and lipid metabolism. However, the evidence regarding the association between adiponectin and diabetes mellitus in obese dogs is sparse. The aim of this study is to investigate the associations of adiponectin with the risk of diabetes mellitus in obese dogs on the basis of a prospective cohort study. MATERIALS AND METHODS: Serum adiponectin levels in obese dogs recruited from three small animal hospitals between 2015 and 2018 were measured by ELISA. Electronic health records were used to record the incidence of diabetes mellitus during follow-up for 3 years. RESULTS: A total of 862 dogs were included. Amongst the 862 dogs, 51 developed diabetes. Adiponectin levels were associated with diabetes mellitus after adjusting for sex, age, breed, exercise, body condition score, fasting plasma glucose, serum triglyceride and total cholesterol. When adjusting for sex, age, breed, exercise, body condition score, fasting plasma glucose, serum triglyceride and total cholesterol, the adjusted hazard ratios were 7.83 (95% confidence interval: 2.67 to 30.13) in the lowest adiponectin group and 1.96 (95% CI: 1.10 to 8.55) in the medium adiponectin group relative to that in the highest adiponectin group. The area under a curve of adiponectin's Receiver operating characteristic curve was 0.81 (95% CI: 0.76 to 0.86). CLINICAL SIGNIFICANCE: Low adiponectin is associated with diabetes mellitus and has a high risk of incident diabetes mellitus, implying the potential of adiponectin as a predictive biomarker of diabetes mellitus in obese dogs.

First Report of Colletotrichum fructicola Causing Bitter Rot of Pear (Pyrus bretschneideri) in China.

Pyrus bretschneideri cv. Dangshansuli is the most important commercial Asiatic pear cultivar worldwide. In recent years, a fruit rot disease of unknown etiology have caused considerable fresh market losses in the 'Dangshansuli' production operations in Dangshan county, Anhui Province, China. Fresh market losses typically range from 60 to 90% and in 2008 were estimated at US$150 million. Symptomatic mature 'Dangshansuli' pears were collected from an orchard in Dangshan County in February 2008. A thin section (about 1 mm3) of symptomatic tissue was sterilized in a bleach and placed on potato dextrose agar (PDA) medium for isolation. From all fruit, a single fungus was recovered displaying gray-white dense aerial mycelium. Identical fungi were isolated from six additional symptomatic 'Dangshansuli' pears collected from other orchards in the county. Pathogenicity tests using one isolate (DS-0) were conducted in triplicate by placing 4 mm diameter discs from 7-day-old PDA plates onto the mature 'Dangshansuli' pear fruit that were incubated in an incubator at 25°C with a 12-h photoperiod for 30 days. An equal number of noncolonized PDA inoculations were included as a control. Isolate DS-0 caused symptoms similar to those in the field within 7 days and complete collapse of cortical tissues within 30 days. No symptoms were observed on control fruit. Round brownish lesions with a diameter of about 3 cm on inoculated fruit was populated by sunken, rotiform acervuli on which numerous, colorless, oblong single cell shape conidia with width/length of 6 × 20 µm were produced. A comparison of morphology and sequence analysis of the ribosomal internal transcribed spacer (ITS) regions in pre- and post-inoculation cultures from inoculated fruit confirmed the presence DS-0. To further characterize DS-0, aliquots of extracted genomic DNA from the fungus were subjected to PCR amplification and sequencing of seven gene regions from the ITS, actin (ACT), ß-tubulin 2 (TUB2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), manganese-superoxide dismutase (SOD2), chitin synthase (CHS-1), and calmodulin (CAL), using the primers listed by Weir et al (4), except for the primer pair of ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3') for ITS amplification, and SODglo2-R (5'-TAGTACGCGTGCTCGGACAT-3') and SODglo2-R (5'-TAGTACGCGTGCTCGGACAT-3') for TBU2 amplification. Two or three clones of PCR products of each gene were sequenced and compared (GenBank Accession Nos. KC410780 to KC410786) to published data at http://www.cbs.knaw.nl/colletotrichum . The result indicated that DS-0 shared the highest similarity of 99.91% with Colletotrichum fructicola, corroborating numerous reports of Colletotrichum spp. causing bitter rot of pear on P. pyrifolia (1,2,3,4). C. fructicola was only recently reported as causing bitter rot of P. pyrifolia (4) and to our knowledge, this is the first report of C. fructicola causing bitter rot of P. bretschneideri, which will help producers select the best management practices for this devastating disease. References: (1) P. F. Cannon et al. Stud. Mycol. 73:181, 2012. (2) N. Tashiro et al. J. Gen. Plant Pathol. 78:221, 2012. (3) G. K. Wan et al. Mycobiology 35:238, 2007. (4) B. S. Weir et al. Stud. Mycol. 73:115, 2012.

First Report of Plum bark necrosis stem pitting-associated virus in Stone Fruit Trees in China.

Plum bark necrosis and stem pitting disease was first observed on a 'Black Beaut' plum (Prunus salicina Lindl.) in the United States in 1986 and later is several other countries. Plum bark necrosis stem pitting-associated virus (PBNSPaV; genus Ampelovirus, family Closteroviridae), the putative causal agent of the disease, infects many stone fruit species and causes decline, gummosis, flattening of scaffold branches, and stem necrotic pits in some diseased trees (1,3). An investigation of the incidence of PBNSPaV on stone fruit trees in China was conducted during 2009 and 2010. Leaf samples were collected from 47 trees, including peach (P. persica L. Batsch), nectarine (P. persica L. var. nucipersica Schneider), plum (P. domestica L.), ornamental plum (P. cerasifera Ehrb), sweet cherry (P. avium L.), and flowering cherry (P. serrulata L.), grown in Hubei, Henan, and Shandong provinces in central and northern China. Most of sampled trees showed trunk gummosis or stem pitting. The presence of PBNSPaV was tested by reverse transcription (RT)-PCR using primer set PBN195F/PBN195R (5'-CTGGTCTTCCTGCTACTCCTT-3'/5'-AAGCCCACAATCTCAGAGCG-3') designed for the detection of the coat protein (CP) gene of the virus. Total RNA was extracted from leaves using a CTAB protocol reported by Li et al. (2). Products of the expected size of 190 bp were amplified from 20 samples, including seven cultivated peach, four ornamental peach, one nectarine, two plum, one ornamental plum, three sweet cherry, and two flowering cherry samples. All trees positive for PBNSPaV showed stem pitting symptoms on the base of the trunk. To further confirm these results, a 590-base region of the heat shock protein 70 homolog (HSP70h) gene was amplified by RT-PCR using primers HSP-P1/HSP-P2 (5'-GGAATTGACTTCGGTACAAC-3'/5'-TCGAAAGTACCACCACCGAA-3'). Amplicons of the expected size were cloned into the vector pMD18-T (TaKaRa, Dalian, China) and sequenced by Genscript Corp. (Nanjing, China). Sequences of 18 PBNSPaV isolates were deposited in GenBank with Accession Nos. JF810177-JF810194. Sequence comparisons showed that the partial HSP70h gene from the Chinese PBNSPaV isolates shared 82.2 to 100% nucleotide (nt) and 94.0 to 100% amino acid (aa) similarities between them and 83.6 to 99.1% nt and 94 to 100% aa similarities with the corresponding region of the other PBNSPaV isolates deposited in GenBank. In July 2010, peach GF305 seedlings were inoculated by side grafting with budwoods from two PBNSPaV-positive ornamental peach plants. In June 2011, grooving symptom was observed on the stems of the seedlings and the virus was detected by RT-PCR. The results further confirmed PBNSPaV infection in China. These results show that PBNSPaV and the associated disease occur in main cultivated and ornamental Prunus species in China. Given the importance and the devastating symptoms of the disease, it is important to prevent virus spread by using virus-tested propagation materials. References: (1) M. Al Rwahnih et al. Arch. Virol. 152:2197, 2007. (2) R. Li et al. J. Virol. Methods 154:48, 2008. (3) D. B. Marini et al. Plant Dis. 86:415, 2002.

First Report of Citrus exocortis viroid from Grapevine in China.

Citrus exocortis viroid (CEVd) can induce bark scaling, dwarfing, leaf epinasty, and fruit yield loss in susceptible hosts. In citrus, CEVd is reported from around the world, but in grape, it is reported from fewer locations (Australia, Brazil, California, and Spain [1]). In 2009, leaves were collected from 40 grapevines (of several different cultivars and species) from Henan, Hubei, Shandong, and Liaoning provinces, China. Total RNA or double-stranded RNA was extracted from the leaves by a described method (3) and subjected to reverse transcription with a random primer (Takara, Dalian, China) and then PCR with primer CEV-AM3 and CEV-AP3 (2). Results showed that the target DNA fragments of 372 bp long were amplified only from the symptomless leaves collected from two grapevines of cv. White Rose grown for approximately 26 years within a small garden in Hubei Province. Amplified products were recovered and cloned into pMD18-T (Takara) and 10 positive clones of each isolate were sequenced and aligned. For both isolates, 20% of the clones represented the same variant (CEVd-hn-g-1; GenBank Accession No. GU592444). It showed a max identity of 94 to 99% with the variants (GenBank Accession Nos. Y00328.1 and DQ471996.1) from grape registered in NCBI, 91 to 100% (GenBank Accession Nos. DQ431993.1 and DQ831485.1) from citrus, 91 to 98% (GenBank Accession Nos. EF488068.1 and EF488050.1) from broad bean, and 89 to 94% (GenBank Accession Nos. AY671953.1 and S67446.1) from tomato. To our knowledge, this is the first report of CEVd from grape in China. References: (1) M. Eiras et al. Fitopatol. Bras. 31:440, 2006. (2) H. J. Gross et al. Eur. J. Biochem. 121:249, 1982. (3) W. X. Xu et al. J. Virol. Methods 135:276, 2006.

First Report of Onion yellow dwarf virus in Allium chinense in China.

Allium chinense (G. Don) is an economically important vegetable that has been considered to be of Asian origin (2). In April of 2007, plants of cultivated A. chinense showing mosaic, chlorotic streak, twist, and crinkle on leaves were collected from fields at Jiangxia, a suburban district of Wuhan, Hubei Province, China, and transplanted into pots in a greenhouse. Ultrathin sections of diseased leaves were observed under an electron microscope. Pinwheel or cylindrical inclusions, typical of a potyvirus infection, were observed. Filamentous virus particles (820 nm long) were also observed in the crude extract from the same plant. To identify the virus species, viral RNAs were extracted from partially purified virion preparation (3) and used as a template. First-strand cDNA was synthesized with M4-T (5'-GTT TTC CCA GTC ACG AC (T)15-3') as a complementary primer. The 3'-terminus of viral RNA was amplified using primer M4 (5'-GTT TTC CCA GTC ACG AC-3') in combination with a degenerate primer (5'-GGXAAYAAYAGY GGX CARCC-3') that was specific for potyviruses (1). Amplified products were cloned and nine clones were sequenced. Sequence analysis showed that the size of the amplified fragment, excluding the poly A tail, was 1,625 bp, which had the typical characteristics of the 3'-terminus of the potyvirus genome, including the partial NIb gene (636 bp) and the complete coat protein (CP gene; 771 bp). The 1,625-bp sequence from A. chinense (Genbank Accession No. FJ765739) had the highest identity at the nucleotide level with sequences of Onion yellow dwarf virus (OYDV), ranging from 76% (Genbank Accession Nos. AB219833 and AB219834) to 99% (Genbank Accession No. AJ409313). The CP gene had 88% (Genbank Accession Nos. AB219833 and AB219834) to 99% (Genbank Accession Nos. AJ409313 and AJ409310) identity at the amino acid level with corresponding regions of known OYDV isolates from other hosts. Until now, only Scallion mosaic virus (ScaMV), in the genus Potyvirus, had been detected from A. chinense (1); however, OYDV was reported in rakkyo, the same species as Chinese scallion, in Japan (4). To our knowledge, this is the first report of OYDV infecting A. chinense in China. References: (1) J. Chen et al. Arch. Virol. 147:683, 2002. (2) L. K. Mann and W. T. Stearn. Econ. Bot. 14:69, 1960. (3) T. Nagakubo et al. Phytopathology 84:640,1994. (4) I. Sako et al. Ann. Phytopathol. Soc. Jpn. 57:65, 1991.