[Preclinical study of T cell receptor specifically reactive with KRAS G12V mutation in the treatment of malignant tumors].

OBJECTIVE: KRAS gene is one of the most common mutations of proto-oncogenes in human tumors, G12V is one of the most common mutation types for KRAS. It's challenging to chemically acquire the targeted drug for this mutation. Recent studies reported that this mutation peptides can form a neoepitope for T cell recognition. Our study aims to clone the T cell receptor (TCR) which specifically recognizes the neoepitope for KRAS G12V mutation and constructs TCR engineered T cells (TCR-T), and to investigate if TCR-Ts have strong antitumor response ability. METHODS: In this study, tumor infiltrating lymphocytes were obtained from one colorectal cancer patient carrying KRAS G12V mutation. Tumor-reactive TCR was obtained by single-cell RT-5' rapid-amplification of cDNA ends PCR analysis and introduced into peripheral blood lymphocytes to generate TCR-Ts. RESULTS: We obtained a high-affinity TCR sequence that specifically recognized the HLA-A*11:01-restricted KRAS G12V8-16 epitope: KVA11-01. KVA11-01 TCR-T could significantly kill various tumor cells such as PANC-1, SW480 and HeLa (overexpressing HLA-A*11:01 and KRAS G12V), and secreting high levels of interferon-γ (IFN-γ). Non-specific killing experiments suggested KVA11-01 specifically recognized tumor cells expressing both mutant KRAS G12V and HLA-A*11:01. In vivo assay, tumor inhibition experiments demonstrated that infusion of approximately 1E7 KVA11-01 TCR-T could significantly inhibit the growth of subcuta-neously transplanted tumors of PANC-1 and HeLa (overexpressing HLA-A*11:01 and KRAS G12V) cells in nude mice. No destruction of the morphologies of the liver, spleen and brain were observed. We also found that KVA11-01 TCR-T could significantly infiltrate into tumor tissue and had a better homing ability. CONCLUSION: KVA11-01 TCR-T cells can effectively target a variety of malignant tumor cells carrying KRAS G12V mutation through in vitro and in vivo assay. KVA11-01 TCR-T cells have excellent biological activity, high specificity of target antigen and homing ability into solid tumor tissue. KVA11-01 TCR-T is expected to be an effective treatment for patients with KRAS G12V mutant solid malignancies.

[Development and clinical application of the health education knowledge assessment questionnaire for gout].

OBJECTIVE: To develop and test the health education knowledge assessment questionnaire for gout and to investigate the understanding degree of health education knowledge in patients with gout. METHODS: From June 2019 to June 2019, 150 cases of gout patients were enrolled.According to the literature review and the healthy education requirements of gout patients, the framework of education knowledge system was preliminarily formed.The pre-test questionnaire was obtained through two rounds of he Delphi technique.A survey of 150 patients with gout was carried out.The analysis and selection of the questionnaire were based on the coefficient of variation, the analysis of determination value, the correlation coefficient of the items and the total scores, and the exploratory factor analysis.In this study, we evaluated the reliability of internal consistency, semi-reliability.Validity test mainly included content validity and construct validity.In addition, a total of 150 patients with gout in our hospital and outpatient gout were selected to investigate the understanding degree of health education knowledge from June 2019 to December 2019. RESULTS: The significance of the first level index of the questionnaire was 3.83-5.00, the secondary index was 3.00-4.83, and the variation coefficient of each item was 0.31-1.23, and the critical ratio(CR) value of each item in this questionnaire was 3.168-8.333.The Pearson correlation coefficient of each item and the total score of this study was 0.319-0.544.After exploratory factor analysis, some topics were deleted in four dimensions, and there were 16 entries in the final questionnaire.Cronbach' s α coefficient of this questionnaire was 0.715, split-half reliability Spearman-Brown coefficient was 0.785, and retest reliability coefficient was 0.729. The correlation coefficient between each factor of this questionnaire and the total questionnaire was 0.300-0.701, and the correlation coefficient between each item of the questionnaire and each factor was 0.402-0.732, all P < 0.05. The results were statistically significant. By questionnaire investigation, the total score of questionnaire was (6.85±3.22), the score of disease-related knowledge was (2.03±1.24), the score of dietary guidance was (1.53± 1.06), the score of exercise guidance was (2.19±1.24), the score of medication guide was (1.24±1.22). CONCLUSION: The Health Education Knowledge Assessment Questionnaire For Gout has a good reliability and validity for measuring related content, the compilation process is scientific and the content is comprehensive, which can be further applied in clinic.The understanding degree of health education knowledge is low in Chinese patients with gout, and knowledge of gout medicine is lacking especially.

[The application of next-generation sequencing in the diagnosis of Pneumocystis pneumonia in non-HIV infected patients].

Objective: To evaluate the clinical value of next-generation sequencing in the diagnosis of Pneumocystis pneumonia in non-HIV infected patients. Methods: A retrospective study was conducted on the diagnosis and treatment of Pneumocystis pneumonia in 5 non-HIV patients in the Fourth Medical Center of the General Hospital of the PLA from September 1, 2017 to September 1, 2018. Next-generation sequencing of BALF were compared with the traditional laboratory microbiological test, and the advantages of the next-generation sequencing in the diagnosis of Pneumocystis pneumonia in non-HIV infected patients were analyzed. Results: There were 3 males and 2 females, with a mean age (48±6) years. Three patients had membranous nephropathy, a patient had tuberculous meningitis, and a patient had esophageal cancer after radiotherapy and chemotherapy. All patients had glucocorticoid medication history before. The clinical manifestations were fever, cough and dyspnea. The chest CT mainly showed bilateral lung ground glass shadows. All the results of 1, 3-ß-D-glucan test were more than 1 000 ng/L. Bronchoalveolar lavage was performed in the 5 cases, and Pneumocystis cysts were found in 1 BALF by Gomori's methenamine silver nitrate staining, and the DNAs of Pneumocystis and human herpesvirus were detected in 5 BALFs by next-generation sequencing. All patients were treated with sulfamethoxazole/trimethoprim (orally, 1.44 g, q8 h) for 23 to 72 days (median 33 days), and with ganciclovir(Ⅳ, 250 mg q12 h) for 6 to 22 days (median 15 days). The chest CT manifestations and symptoms were improved after treatment, without death. Conclusions: The next-generation sequencing of BALF is more specific and sensitive in the diagnosis of Pneumocystis pneumoniae in non-HIV patients. It is faster, more comprehensive and more accurate than the traditional laboratory test, and could be widely used as a PCP diagnosis technique.

[Efficacy of hierarchical medical mode path management on the continuous treatment for chronic wound patients].

Objective: To explore the efficacy of hierarchical medical mode path management on the continuous treatment for chronic wound patients. Methods: From June 2017 to September 2018, the clinical data of 101 patients with chronic wounds who were just discharged from Department of Wound Repair of the Affiliated Hospital of Jiangnan University (hereafter referred to as the author's affiliation) and still needed continuous treatment, meeting the inclusion criteria, were analyzed with the method of retrospective cohort study. Based on the management method of continuous treatment after discharge, the patients were divided into path management group (52 patients, 27 males and 25 females, aged (44±6) years, 57 wounds) and conventional management group (49 patients, 26 males and 23 females, aged (45±6) years, 53 wounds). The patients in path management group were carried out with full-path management under hierarchical medical mode, and the patients in conventional management group were carried out with conventional continuous treatment management. The discharge time from the author's affiliation was the time before continuous treatment management (hereafter referred to as before management), and 12 weeks post discharge was the time after continuous treatment management (hereafter referred to as after management). The depression and anxiety of patients in two groups were assessed by Hamilton Depression Scale 24 item version (HAMD-24) and Self-rating Anxiety Scale (SAS), and the positive ratios of depression and anxiety were calculated. The effective rates of wound treatment, times of debridement and dressing change, and treatment cost during the continuous treatment management period were counted. Data were statistically analyzed with two independent sample t test, chi-square test, Fisher's exact probability test, and Wilcoxon rank-sum test. Results: (1) Before management, the HAMD-24 and SAS scores of patients in two groups were similar. After management, the HAMD-24 and SAS scores of patients in path management group were significantly lower than those of conventional management group (t=4.341, 3.840, P<0.01). Before and after management, the positive rates of depression and anxiety of patients in two groups were similar. (2) After management, the effective rate of wound treatment of patients in path management group was 98.25% (56/57), which was significantly higher than 86.79% (46/53) of conventional management group (χ(2)=5.341, P<0.05). (3) During the management, the times of debridement and dressing change in patients of path management group was 20±4, which was significantly less than 27±7 of conventional management group (t=5.833, P<0.01). (4) During the management, the treatment cost of patients in path management group was (2 479±213) yuan, which was significantly less than (5 215±326) yuan of conventional management group (t=50.185, P<0.01). Conclusions: In the continuous treatment of chronic wound patients, the full-path management under hierarchical medical mode can improve the effective rate of wound treatment, reduce the times of debridement and dressing change and treatment cost, and improve their psychological state.

Downregulated long non-coding RNA TRPM2-AS inhibits cisplatin resistance of non-small cell lung cancer cells via activation of p53- p66shc pathway.

OBJECTIVE: Non-small cell lung cancer (NSCLC), as an ordinary malignant tumor, presents with high death rate and poor prognosis. Few literatures have explored the association between NSCLC development and lncRNAs expression. This study focuses on the important role of a novel lncRNA TRPM2-AS in the development of chemo-resistance in NSCLC. MATERIALS AND METHODS: The expression level of lncRNA TRPM2-AS was identified by using qRT-PCR assay. The apoptosis rate and the alteration of the cell cycle were detected by the flow cytometric analysis. Cell Counting Kit-8 assay (CCK8) was utilized for detecting chemo-sensitivity of the cisplatin-resistant A549/DDP cells. The p53 and p66shc protein levels were detected by Western blotting assay. RESULTS: A549/DDP cells presented remarkably higher expression of lncRNA TRPM2-AS than paired A549 cells. Moreover, re-sensitization to cisplatin was seen in A549/DDP cells after lncRNA TRPM2-AS knockdown. On the contrary, the sensitivity of lncRNA TRPM2-AS-overexpressed A549 cells to cisplatin decreased obviously when compared with the control. Furthermore, downregulated lncRNA TRPM2-AS induced cell apoptosis and altered cell cycle distribution through activating the p53-p66shc pathway. CONCLUSIONS: We suggest that lncRNA TRPM2-AS participates in the resistance of NSCLC cells to cisplatin, which may provide a new therapeutic target of NSCLC.

Metabolic profiling reveals new serum biomarkers of lupus nephritis.

Metabolomics has been applied to explore altered metabolite profiles in disease and identify unique metabolic signatures specific to certain pathologies. The aim of the current study is to characterize the metabolic profile of patients diagnosed with lupus nephritis (LN) and explore new insights into underlying disease processes. A metabolomic approach using ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UPLC-HRMS) was developed in serum samples from 32 LN patients, 30 idiopathic nephrotic syndrome (INS) patients and 28 healthy controls (HCs). Potential biomarkers were screened from orthogonal projection to latent structures discriminate analysis (OPLS-DA) and further evaluated by receiver operating characteristic analysis (ROC). A total of 14 potential biomarkers were screened and tentatively identified for LN patients compared to HCs. Compared to HCs and INS patients, the LN patients had increased serum levels of sorbitol and glycocholic acid metabolites and decreased levels of cortisol, creatinine and L-aspartyl-L-phenylalanine. A panel of three metabolomics (theophylline, oxidized glutathione and capric acid) was identified as biomarkers of LN with a sensitivity of 87.50% and a specificity of 67.86% using ROC analysis. Our results suggest that UPLC-HRMS based quantification of circulating metabolites was a useful tool for identification of biomarkers with the ability to segregate LN patients from INS patients and HCs. The potential biomarkers indicated that the LN metabolic disturbance may be closely associated with inflammation injury, oxidative stress and phospholipid metabolism.

Evaluation of a loop-mediated isothermal amplification assay based on hrpZ gene for rapid detection and identification of Pseudomonas syringae pv. lachrymans in cucumber leaves.

AIMS: Cucumber angular leaf spot caused by Pseudomonas syringae pv. lachrymans (Psl) is an important and destructive disease worldwide, and no effective technique has been developed for the control of the pathogen. Detection of infection or latent in cucumber plants is critical to evaluate disease progress and strengthening management to avoid a serious epidemic in the fields. In this paper, we developed a rapid and sensitive method for detection of Psl using an isothermal method known as loop-mediated amplification (LAMP). METHODS AND RESULTS: A set of six primers was designed to amplify the gene coding for the hrpZ, and conditions for detection were optimized to complete in 60 min at 67°C, and the amplification were confirmed through gel electrophoresis or visually inspected using calcein stain. The specificity of LAMP primers set was widely validated on Psl and nontarget strains. In sensitivity testing, LAMP allowed detection as low as 104  CFU per ml bacterial cells without DNA extraction. The novel method was also applied for detecting Psl in infected cucumber leaves, and even the early onset of disease can be detected by the assay. CONCLUSIONS: This study confirmed that the novel developed LAMP assay is an easy, rapid and sensitive method for the detection of Psl in infected leaves. SIGNIFICANCE AND IMPACT OF THE STUDY: The method is suitable for direct detection of Psl without strain enrichment and complex DNA extraction from samples in the field, and hence it has the capability to be used for on-site disease diagnosis and field surveys.

Role of HOTAIR long noncoding RNA in metastatic progression of lung cancer.

OBJECTIVE: Lung cancer, the primary cause of cancer-related death worldwide, imperatively requires new therapeutic approaches that target the molecular regulatory machinery. Even though it has been reported that long noncoding RNAs are involved in different cancer types, limited data are available on the expression and function of long noncoding RNAs in lung cancer metastasis. The major objective of the current study was to profile expression and function of six long noncoding RNAs in five matched pairs of primary lung cancer and lymph node metastatic tissues. MATERIALS AND METHODS: The study protocol was approved by the Institutional Review Board of the Chinese PLA General Hospital, China. All patients enrolled in the study provided signed informed consent. RESULTS: Among the tested lncRNAs, HOTAIR and NEAT1 were most highly expressed in lymph node metastasis. However, only HOTAIR was subsequently found to be involved in lung cancer cell motility and invasion, as assessed by in vitro migration and invasion assay. Finally, our experiments suggest that HOTAIR promoted gelatinase activity in these cells. CONCLUSIONS: Our data indicates that HOTAIR is overexpressed in metastatic lung cancer tissue, which is prospectively associated with the ability of HOTAIR to promote lung cancer cell motility and invasion.

First Report of Pseudomonas cichorii Causing Leaf Spot of Vegetable Sponge Gourd in China.

A suspect bacterial leaf spot on vegetable sponge gourd (Luffa cylindrical (L.) Roem.) was found in a commercial greenhouse in Pi County, Chengdu City, Sichuan Province, China, in February 2011. Approximately 20 to 30% of plants were affected, causing serious economic loss. Symptoms occurred only on seedlings and consisted of water-soaked, irregularly shaped, black lesions on the surface and margins of cotyledons. A bacterium was consistently isolated on nutrient agar from diseased leaf tissues that had been surface disinfected in 70% ethyl alcohol for 30 s. The bacterium produced small gray colonies with smooth margins, was gram negative, fluoresced on King's B medium, and showed pectolytic activity when inoculated on potato slices. The partial sequences of 16SrRNA gene (1,377 bp) of the bacterium (GenBank Accession No. KC762217), amplified by using universal PCR primers 16SF (5'-AGAGTTTGATCCTGGCTCAG-3') and 16SR (5'-GGTTACCTTGTTACGACTT-3'), shared 100% similarity with that of Pseudomonas cichorii (GenBank Accession No. HM190228). The vegetable sponge gourd isolate was also identified by using the Biolog Microbial Identification System (version 4.2, Biolog Inc., Hayward, CA) as P. cichorii with the following characteristics (1): negative for arginine dihydrolase, gelatin liquefaction, and N2 production. Positive reactions were obtained in tests for catalase, oxidase, potato rot, utilization of melibiose, and mannitol. Tests were negative for utilization of sucrose, trehalose, D-arabinose, raffinose, cellobiose, and rhamnose. A pathogenicity test was conducted on 4-week-old vegetable sponge gourd plants by spray-inoculation with 108 CFU/ml sterile distilled water on the leaves of 15 vegetable sponge gourd plants and by needle puncture on the stems of 15 other plants with P. cichorii, respectively. Control plants were misted with sterile distilled water or punctured on the stem with a clean needle. Plants were placed in a greenhouse maintained at 28 ± 2°C with relative humidity of 80 to 85%. Symptoms, the same as seen on the original diseased plants, developed after 7 to 10 days on inoculated plants. Control plants remained healthy. The bacterium was readily re-isolated from inoculated plants and identified as P. cichorii using P. cichorii-specific primer hrpla/hrp2a (1). To our knowledge, this is the first report of P. cichorii causing disease on commercially grown vegetable sponge gourd in China. This new finding will provide the basis for developing resources for diagnostics and management, including screening varieties for resistance. References: (1) S. Mazurier et al. J. FEMS Microbiol. Ecol. 49:455, 2004. (2) N. W. Schaad et al., eds. Laboratory Guide for Identification of Plant Pathogenic Bacteria, 3rd ed. APS Press, St. Paul, MN, 2001.

First Report of Myrothecium roridum Causing Leaf Spot on Zantedeschia aethiopica in China.

Zantedeschia aethiopica (L.) Spreng. (calla lily), belonging to family Araceae, is a popular ornamental plant in China. In the summer of 2010, leaves of calla lily with typical symptoms of necrotic lesions were observed in a commercial glasshouse in Beijing, China (116°20' E, 39°44' N). The initial symptoms were circular to subcircular, 1 to 3 mm, and dark brown lesions on the leaf lamina. Under high humidity, lesions expanded rapidly to 5 to 10 mm with distinct concentric zones and produced black sporodochia, especially on the backs of leaves. Later, the infected leaves were developing a combination of leaf lesions, yellowing, and falling off; as a result, the aesthetic value of the plant was significantly impacted. Leaf samples were used in pathogen isolation. Symptomatic leaf tissues were cut into small pieces and surface sterilized with 70% ethanol for 30 s and then in 0.1% mercuric chloride solution for 1 to 3 min. After being washed in sterile distilled water three times, the pieces were plated on potato dextrose agar (PDA) and incubated at 25°C in darkness for 7 days (5). Initial colonies of isolates were white, floccose mycelium and developed dark green to black concentric rings that were sporodochia bearing viscid spore masses after incubating 5 days. Conidiophores branched repeatedly. Conidiogenous cells were hyaline, clavate, and 10.0 to 16.0 × 1.4 to 2.0 µm. Conidia were hyaline, cylindrical, both rounded ends, and 6.0 to 8.2 × 1.9 to 2.4 µm. Morphological characteristics of the fungus were consistent with the description of Myrothecium roridum Tode ex Fr. (3,4). To confirm the pathogenicity, three healthy plants of calla lily were inoculated with a conidial suspension (1 × 106 conidia per ml) brushed from a 7-day-old culture of the fungus. Control plants were sprayed with sterile water. The inoculated plants were individual with clear plastic bags and placed in a glass cabinet at 25°C. After 7 days, all inoculated leaves developed symptoms similar to the original samples, but control plants remained disease free. Re-isolation and identification confirmed Koch's postulates. For molecular identification, genomic DNA of a representative isolate (MTL07081001) was extracted by modified CTAB method (1), and the rDNA-ITS region was amplified by using primers ITS1 (5-TCCGTAGGTGAACCTGCGG-3) and ITS4 (5-TCCTCCGCTTATTGATATGC-3). The 465-bp amplicon (GenBank Accession No. KF761293) was 100% identity to the sequence of M. roridum (JF724158.1) from GenBank. M. roridum has an extensive host range, covering 294 host plants (2). To our knowledge, this is the first record of leaf spot caused by M. roridum on calla lily in China. References: (1) F. M. Ausubel et al. Current Protocols in Molecular Biology. John Wiley & Sons Inc, New York, 1994. (2) D. F. Farr and A. Y. Rossman, Fungal Databases. Syst. Mycol. Microbiol. Lab., ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , October 2013. (3) M. T. Mmbaga et al. Plant Dis. 94:1266, 2010. (4) Y. X. Zhang et al. Plant Dis. 95:1030, 2011. (5) L. Zhu et al. J. Phytopathol. 161:59, 2013.

Endothelial nitric oxide synthase and nicotinamide adenosine dinucleotide phosphate oxidase p22phox gene (C242T) polymorphisms and systemic lupus erythematosus in a Chinese Population.

Genetic polymorphisms in the endothelial nitric oxide synthase (eNOS) and nicotinamide adenosine dinucleotide phosphate (NADPH) oxidase p22phox are linked with the expression and/or progression of vascular disease. We hypothesized that these polymorphisms may influence the development and/or progression of systemic lupus erythematosus (SLE), given their linkage with vascular disease. DNA from patients with SLE (n = 90) and their age- and sex-matched controls (n = 86) from The Second Xiangya Hospital of Central South University was assessed for eNOS and NADPH oxidase p22phox polymorphisms. These polymorphisms were examined by restriction fragment length polymorphism-polymerase chain reaction. The allele frequency of the NADPH oxidase p22phox gene C242T polymorphisms significantly varied between the SLE patients and the controls. We found no association of the eNOS polymorphism with the development of renal disease. These results indicated that the etiology of patients with SLE is associated with NADPH oxidase p22phox gene C242T polymorphisms. There was no significant increased risk of SLE associated with eNOS polymorphisms in the Chinese population.

First Report of Crown Rot of Grafted Cucumber Caused by Fusarium solani in China.

Grafting has been widely and effectively used in cucumber (Cucumis sativus) cultivation for approximately 30 years in China to avoid Fusarium wilt caused by Fusarium oxysporum Schl. f. sp. cucumerinum Owen. In greenhouses, 90% of cucumbers are grafted onto pumpkin (Cucurbita moschata) rootstock. However, in March 2009, a severe crown rot causing yellowing and wilting of the leaves was observed on grafted cucumber in a large number of greenhouses in Lingyuan, western Liaoning Province in China. Symptoms consisted of dark brown, water-soaked lesions and a dense, white mycelial mat at the base of the stem. Lingyuan is the largest district for cucumber cultivation using grafting techniques in solar greenhouses in China. In 30 surveyed greenhouses in Sanshijiazi Village in the city of Lingyuan, the incidence of affected plants ranged from 10 to 40%, which caused serious economic losses. Fusarium spp. were isolated from the surface-sterilized basal stems of symptomatic plants on potato dextrose agar and incubated on potato sucrose agar for 4 days at 25°C. Colonies of the isolates produced a brown pigmentation and sparse, aerial mycelia, with a cream color on the underside. Conidiophores were elongated and branched or unbranched. Microconidia were abundant, hyaline, ellipsoid to ovoid, and 6 to 14 × 2.5 to 3.5 µm. Macroconidia were cylindrical, abundant, mostly two to six septate, 22 to 63 × 3.2 to 5.0 µm, with the apical cell rounded and blunt, and the basal cell rounded. On the basis of morphological characteristics, the fungus was identified as F. solani (C. Booth). For confirmation, the internal transcribed spacer region of rDNA was amplified and sequenced. A 449-bp sequence shared 99% homology with that of a F. solani GenBank accession previously reported from Japan (No. AF150473.1). The new sequence was deposited in GenBank (Accession No. HM015882). Pathogenicity of three isolates was determined in two experiments using different methods of inoculation. In one, 30 seedlings of pumpkin (C. moschata) with one true leaf each were inoculated by dipping their roots in a suspension of 106 spores ml-1, while control plants were mock inoculated with sterile water. Plants were then potted in a sterile mix of peat moss and vermiculite (2:1 vol/vol). In the other, pregerminated pumpkin seeds were sown in the same medium with a conidial suspension added at a rate of 106 spores ml-1, while other seeds were sown in sterile soil as controls. Plants for both experiments were maintained in a greenhouse at 25°C. Twelve days after inoculation, inoculated plants in both experiments showed a cortical rot on the crown and stem base with a brown, water-soaked appearance. Twenty-one days later, inoculated plants developed wilting and yellowed leaves. Disease incidence was 100%. No symptoms occurred on the control plants. Both experiments were repeated once with the same results. The pathogen was recovered from symptomatic tissue, confirming Koch's postulates. F. solani has been previously reported to cause root rot on cucurbit in California (2) and crown rot on grafted cucumber in the Netherlands (1). To our knowledge, this is the first report of crown rot of grafted cucumber caused by F. solani in China. References: (1) L. C. P. Kerling and L. Bravenboer. Neth. J. Plant Pathol. 73:15, 1967. (2) T. A. Tousson and W. C. Snyder. Phytopathology 51:17, 1961.

First Report of Myrothecium Leaf Spot of Common Bean in China Caused by Myrothecium roridum.

Common bean (Phaseolus vulgaris L.) is an economically important crop in China. In June 2008, a new foliar disease was observed on beans in Shunyi District, Beijing, China. The disease occurred in approximately 15% of the plants in a commercial field. Leaf spots were circular to irregular, reddish brown, zonate, and 8 to 20 mm in diameter. Black sporodochia with white tuffs were present on older lesions and black spore masses were present in moist weather. Ten isolates recovered from lesions produced white, floccose colonies and spore masses after 4 days on potato dextrose agar. The rod-shaped, hyaline conidia had rounded ends and averaged 6.8 × 2.5 µm. All characteristics were consistent with the description of Myrothecium roridum Tode ex Fr. (1). The internal transcribed spacer regions of one isolate were sequenced and deposited in GenBank (Accession No. GQ 381291). Sequences of the isolate from bean in China were 98% similar to sequences of M. roridum in GenBank. To determine pathogenicity, 30 healthy seedlings of common bean were inoculated by spraying a 1 × 105 conidia ml-1 suspension of M. roridum onto the foliage. Ten seedlings were sprayed with sterile water and served as controls. Plants were kept in a humid chamber at 27°C overnight and then placed in a growth chamber. After 6 days, the symptoms described above were observed on leaves in all inoculated plants, whereas symptoms did not develop on the control plants. The pathogen was reisolated from inoculated leaves, fulfilling Koch's postulates. There is one report of M. roridum on soybean in Korea (2). To our knowledge, this is the first report of Myrothecium leaf spot on common bean in China. References: (1) M. Fitton et al. CMI Mycol. Pap. No. 253, 1970. (2) K. J. Yum et al. Plant Pathol. J. 6:313, 1990.

First Report of Target Leaf Spot Caused by Corynespora cassiicola on Balsam Pear in China.

Balsam pear (Momordica charantia L.) is an economically important vegetable in China with increasing interest as a medicinal plant. In December of 2006, a new foliar disease caused by Corynespora cassiicola was observed on balsam pear growing in greenhouses in Shouguang City, Shandong Province, China. The disease occurred on 35% or less of the plants. Leaves of affected plants developed off-white halos surrounding circular lesions that were 1 to 5 mm broad. The lesions became dark brown, necrotic with concentric rings, and up to 15 mm in diameter. Severely affected plants eventually wilted and defoliated. Pieces of tissue from the leading edges of lesions were disinfected in 1% NaOCl for 1 min, rinsed in sterile water, and plated on potato dextrose agar. Colonies of the fungus were gray to dark green. Conidiophores were erect and simple, pale brown to brown, and 100 to 450 µm long and 3 to 8 µm wide. Conidia were obclavate to cylindrical, pale olivaceous brown to dark brown, smooth, 35 to 100 × 8 to 12 µm, and were produced in chains. On the basis of these characteristics, the fungus was identified as Corynespora cassiicola (1). The internal transcribed spacer (ITS) region of rDNA was amplified with primers ITS1/ITS4 and deposited in GenBank (Accession No. GQ381292). It was an exact match for a sequence of C. cassiicola previously deposited (Accession No. EU364555). To confirm pathogenicity, 30 1-month-old healthy seedlings of balsam pear were inoculated by spraying a suspension of conidia (1 × 105 conidia per ml) of one isolate of C. cassiicola until runoff. Ten seedlings were sprayed with sterile water as controls. Plants were kept in a humidity chamber at 27°C overnight and then placed in a growth chamber at 27°C. After 7 days, symptoms identical to those described above were observed, while no symptoms developed on the control plants. The pathogen was reisolated from inoculated leaves. C. cassiicola causes foliar diseases on many plants, including tomato, eggplant, soybean, and cucumber (2). There is one report on balsam pear in Korea (3). To our knowledge, this is the first report of target leaf spot caused by C. cassiicola on balsam pear in China. References: (1) M. B. Ellis. CMI Mycol. Pap. No. 65, 1957. (2) M. B. Ellis et al. CMI Mycol. Pap. No. 303, 1971. (3) J. H. Kwon et al. Plant Pathol. J. 21:164, 2005.

Prevalence of diabetic retinopathy among subjects with known diabetes in China: the Beijing Eye Study.

PURPOSE: To evaluate the prevalence of diabetic retinopathy and risk factors among patients with self-reported diabetes mellitus in China. METHODS: The Beijing Eye Study, a population-based study on inhabitants aged 40+ years, included 4439 subjects. Fundus photographs of the worse eye from participants with self-reported diabetes were graded. RESULTS: Fundus photographs ready for evaluation and a filled out questionnaire were available for 4127 (93.0%) subjects. The prevalence of self-reported diabetes was 235/4127 (5.7%). Among the subjects with a self-reported diagnosis of diabetes, diabetic retinopathy was detected on the fundus photographs of 86 (37.1%) subjects, with macular edema in 12 (5.2%) subjects, clinically significant macular edema in 6 (2.6%) subjects, and a vision- threatening stage of the retinopathy in 12 (5.2%) subjects. Diabetic retinopathy was associated with rural region (p=0.004), longer duration of diabetes (p=0.009), use of diabetic medications (p=0.02), and lower education background (p=0.003). CONCLUSIONS: Prevalence of diabetic retinopathy among Chinese patients aged 40+ years with a self-reported diagnosis of diabetes is about 37%, with a vision-threatening stage of the retinopathy detected in 5% of the subjects. About 5.7% of the adult Chinese population report on a known diagnosis of diabetes mellitus, with about 15% of these subjects knowing about the presence of diabetic retinopathy. The frequency of known diabetes mellitus is lower in rural regions than in urban regions, while diabetic retinopathy overall and macular edema among the subjects with known diabetes mellitus were significantly more common in the rural group.

Ocular and systemic factors associated with diabetes mellitus in the adult population in rural and urban China. The Beijing Eye Study.

PURPOSE: To assess ocular and systemic factors associated with diabetes mellitus in the adult population in rural and urban China. METHODS: The Beijing Eye Study 2006, a population-based, cross-sectional cohort study, included 3251 subjects aged 45 years and more (participation rate: 73.2%). Blood samples were available for 2960 (91.0%) subjects. Diabetes mellitus was defined by a fasting plasma glucose concentration >or=7.0 mmol/l or by a self-reported history diagnosis of diabetes. RESULTS: Diabetes mellitus was found in 381 (12.9%) subjects. In binary regression analysis, the presence of diabetes mellitus was significantly associated with body mass index, systolic blood pressure, triglyceride concentrations, intraocular pressure, cylindrical refractive dioptre, presence of arteriolar sheathing, rural vsurban region, lower best-corrected visual acuity, lower high-density lipoprotein level, and lower diastolic blood pressure. It was not statistically associated with age, presence of cataract (nuclear, cortical, or subcapsular), size of the optic disc, neuroretinal rim, alphazone and betazone of peripapillary atrophy, retinal artery and vein diameters, arteriovenous nicking, focal or general narrowing, refractive error, prevalence of glaucoma, and early or late stage of age-related macular degeneration. CONCLUSIONS: In a population-based setting, diabetes mellitus was not associated with optic disc, rim and peripapillary atrophy measurements, retinal vessel diameters, arteriovenous nicking, focal or general retinal artery narrowing, and prevalence of age-related macular degeneration. Although diabetes mellitus was significantly correlated with higher intraocular pressure, it was not associated with glaucoma.