[The cases of tracing the source of patients infected with Omicron variant of SARS-CoV-2 based on wastewater-based epidemiology in Shenzhen].

Wastewater-based epidemiology (WBE) is an emerging discipline, which has been applied to drug abuse tracking and infectious disease pathogen surveillance. During the COVID-19 epidemic, WBE has been applied to monitor the epidemic trend and SARS-CoV-2 variants etc. In order to detect hidden COVID-19 cases and prevent transmission in the community, wastewater surveillance system for monitoring SARS-CoV-2 RNA was developed in Shenzhen. The sewage sampling sites were set up in key places such as the port areas, urban villages and residential communities of Futian, Nanshan, Luohu and Yantian districts. From July 26 to November 30, 2022, a total of 369 sewage sampling sites were set up, covering 1.93 million people. Continuous sampling was carried out for 3 hours in the peak period of water use every day. Sewage virus enrichment and SARS-CoV-2 nucleic acid detection were carried out by polyethylene glycol precipitation method and RT-qPCR, and a positive water sample disposal process was molded. This article aims to introduce the case of source tracing of COVID-19 infected patients based on urban sewage in Shenzhen. The sewage monitoring of Honghu water treatment plant in Luohu District played an early warning role, and the source of infection was traced. In the disposal of positive water samples in Futian South Road, Futian District, the important experience of monitoring point layout was obtained. In the sewage monitoring of Nanshan village, Nanshan District, the existence of occult infection was revealed. Sharing the experience of tracing the source of COVID-19 patients to avoid the spread of COVID-19 in the community based on wastewater surveillance of SARS-CoV-2 RNA in Shenzhen, and summarizing the advantages and application prospects of sewage surveillance can provide new ideas for monitoring emerging or re-emerging pathogens that are known to exhibit gastrointestinal excretion in the future.

[Effect of autologous platelet-rich plasma perfusion on cytokines in uterine drainage fluid of patients with intrauterine adhesions following hysteroscopic adhesiolysis].

Objective: To investigate the effect of autologous platelet-rich plasma (PRP) perfusion on the levels of cytokines in uterine drainage fluid in patients with moderate to severe intrauterine adhesions (IUA) following hysteroscopic adhesiolysis. Methods: Thirty patients with moderate to severe IUA who underwent hysteroscopic adhesiolysis at Beijing Obstetrics and Gynecology Hospital, Capital Medical University from November 2020 to March 2021 were randomly divided into two groups: the PRP group (15 patients with placement of intrauterine-suitable balloons and PRP infusion) and the control group (15 patients with placement of intrauterine-suitable balloons only). For all patients, the channel switch was opened 48 hours after the surgery. The drainage fluid of the uterine cavity was collected using syringes through the proximal end of the drainage channel switch at 24 hours after the surgery and through the drainage channel directly at 48, 72, 96, and 120 hours after the surgery, and the levels of related cytokines including platelet-derived growth factor BB (PDGF-BB), vascular endothelial growth factor A (VEGF-A), insulin-like growth factor 1 (IGF-1) and transforming growth factor-ß1 (TGF-ß1) in the drainage fluid of the uterine cavity were evaluated, respectively. Results: (1) The changes in volumes of uterine cavity drainage fluid: the total drainage fluid volumes of the PRP group and the control group in 120 hours after the surgery were (21.8±2.9) and (22.7±2.7) ml, respectively, and there was no statistically significant difference between the two groups (t=-0.847, P>0.05). No significant differences were found in the volumes of drainage fluid between the two groups at 72, 96, and 120 hours after the surgery (all P>0.05). (2) Variation in cytokine levels in the uterine cavity drainage fluid: ① PDGF-BB: median PDGF-BB levels at 24 and 48 hours after the surgery in the PRP group (6.6 and 9.6 µg/L, respectively) were significantly higher than those in the control group (4.7 and 2.7 µg/L, respectively; all P<0.05). There were no significant differences in PDGF-BB levels between the two groups at 72, 96, and 120 hours after the surgery (all P>0.05). ② VEGF-A: median VEGF-A levels at 24 and 48 hours after the surgery in the PRP group (3.5 and 2.8 µg/L, respectively) were significantly higher than those in the control group (1.6 and 1.2 µg/L, respectively; all P<0.05). There were no significant differences in VEGF-A levels between the two groups at 72, 96, and 120 hours after the surgery (all P>0.05). ③ IGF-1: median IGF-1 level at 48 hours after the surgery in the PRP group was significantly higher than that in the control group (39.5 vs 8.6 µg/L, P<0.05). No significant differences were found in IGF-1 levels at 24, 72, 96, and 120 hours after the surgery between the two groups (all P>0.05). ④ TGF-ß1: There were no significant differences in TGF-ß1 levles between the two groups at 24, 48, 72, 96, and 120 hours after the surgery (all P>0.05). Conclusions: PRP perfusion following hysteroscopic adhesiolysis may increase the levels of PDGF-BB, VEGF-A, and IGF-1 in the uterine cavity drainage fluid, which plays a beneficial role in improving wound microvascular formation, reducing adhesion reformation, and promoting endometrial regeneration and repair.

[Long-term prophylactic therapy combined with on-demand therapy for the control of hereditary angioedema--Comment on the 2021 edition of the international WAO/EAACI guideline for the management of hereditary angioedema].

The 2021 edition of the international World Allergy Organization (WAO)/European Academy of Allergy and Clinical Immunology (EAACI) guideline for the management of hereditary angioedema (HAE) is mainly based on high-quality randomized controlled trials. It provides clinical classification for HAE and offers graded recommendations for on-demand therapy, short-term prophylactic therapy, and long-term prophylactic therapy. Additionally, it provides management strategies for people with different HAE types. This article focused on the interpretation of short-term, long-term prophylactic therapy and on-demand therapy for HAE, supplemented with the latest clinical evidence, aiming to provide references for the long-term management of HAE.

[Investigation on the cognitive level of Mpox among MSM population in Shenzhen and analysis of the influencing factors in 2023].

From June 16 to 30, 2023, men who have sex with men (MSM) who had visited Voluntary Counseling Testing (VCT) clinics in the Luohu, Futian and Nanshan districts of Shenzhen were included in this study to analyze their awareness of Mpox and the influencing factors. The mean age of the 262 MSM was (34.78±8.94) years, with the majority being unmarried (75.2%) and 79.0% confirmed to be infected with HIV. The awareness rates for five primary indicators, current status of Mpox, pathogen and source of infection, mode of transmission, population susceptibility, clinical manifestations and treatment were 68.4%, 84.7%, 60.3%, 87.8%, and 52.5%, respectively. The awareness rates for five secondary indicators, earliest transmission location (44.7%), main mode of transmission (54.2%), role of masks (46.9%), drug accessibility (46.6%), and self-limiting nature (38.2%) were all below 60%. The MSM population in Shenzhen perceived their likelihood of being infected (2.76±1.32) and discriminated against (3.87±1.26) as relatively low. The logistic analysis showed that the high school or vocational school education (OR:3.094, 95%CI:1.180-9.299), college or above education (OR:5.360, 95%CI:2.159-15.501), and higher scores on questions affecting learning or work (OR:2.196, 95%CI:1.409-3.599) were promoting factors for Mpox awareness, while higher scores on questions concerning the possibility of Mpox mortality (OR:0.591, 95%CI:0.432-0.791) was the hindering factor for Mpox awareness.

[The expression of GGCT in the bladder urothelial cell carcinoma and its clinical significance].

Objective: To evaluate the expression and clinical significance of γ-glutamylcyclotransferase (GGCT) in patients with bladder urothelial cell carcinoma. Methods: Immunohistochemical staining for GGCT were performed on tissue sections of 86 patients with bladder urothelial cell carcinoma and 10 normal controls, and the correlations between GGCT and clinicopathological characteristics and the prognosis were analyzed. Results: The positive rate of the expression of GGCT in 86 cases of bladder urothelial cell carcinoma was 61.6% (53/86). GGCT protein was located mainly in cancer cell cytoplasm, and it can be seen in the nucleus of the tumor cells in some cases. The level of GGCT expression was positively related to pathological classification (P<0.001), stage (P=0.020), and tumor size (P=0.025). Immunohistochemical semiquantitative analysis showed that the expression of GGCT in patients with T1 stage of non-muscle invasion bladder urothelial cell carcinoma was significantly higher than that with Ta stage (P=0.034). Kaplan-Meier analysis showed that the expression of GGCT was correlated with the recurrence-free survival in patients with non-muscle invasive bladder cancer, the recurrence-free survival rate was lower in the GGCT positive group (P=0.029). Multivariate COX regression analysis showed that the pathological stage (OR=5.029, P=0.009) and the number of tumors (OR=3.320, P=0.024)were the independent risk factors for recurrence-free survival in patients with early urothelial cell carcinoma of the bladder. Conclusions: The expression of GGCT is significantly increased in bladder urothelial cell carcinoma and is related to the malignant biological behavior and progression of tumor. Patients with GGCT positive early bladder tumor are inclined to recur.

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.

[Effects of acute paraquat poisoning on cognitive ability in humans].

Objective: To explore the effects of acute paraquat poisoning on cognitive function of patients through neuropsychologic test. Methods: In June 2019, 36 patients with acute paraquat poisoning in the emergency department of a provincial hospital in Hebei Province were selected as the case group. 36 healthy individuals were selected as control group. The cognitive function and depressive state were assessed by mini mental state scale, auditory word learning test, digit span test, connection test, Boston Naming Test and geriatric depression scale. Results: The results of Mini-Mental State examination showed that the total score of the case group was lower than that of the control group, the difference was statistically significant (P<0.05) . The results of the Auditory Vocabulary Learning test showed that the scores of delayed recall, clue recall, corrective ability and semantic learning strategies of the case group were significantly lower than those in the control group (P<0.05) . There was no significant difference in the scores of immediate memory between the two groups (P>0.05) . The scores of Digit Span test and Boston Naming test in the control group were higher than those in the case group, the Trail Making test time in the control group was shorter than that in the case group, and the differences were statistically significant (P<0.05) . Conclusion: Acute paraquat poisoning can impair human cognitive ability to a certain extent.

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.

[Analysis of type C behavior in patients with oral lichen planus].

OBJECTIVE: To assess type C behavior in patients with oral lichen planus (OLP) in order to provide basis for clinical prevention, treatment and psychological intervention of OLP. METHODS: Type C behavior scale was used on 85 OLP patients and 85 control patients, who were in accordance with the inclusion criteria, in order to investigate their type C behavior. The scale included 9 items: anxiety, depression, anger, anger toward inside (anger-in), anger toward outside (anger-out), reasoning, domination, optimism, and social support. Scores of the 9 items between OLP patients and control group were calculated under the instruction of the scale and were statistically analyzed, and OLP group was further stratified statistically by sex, reticulate-erosive-ulcerative (REU) pathological type and course of diseases, and the scores of each group were analyzed and compared. RESULTS: Among the 85 OLP patients, there were more females, more non-erosive lesion type, and the most common site for OLP was the buccal mucosa. The scores of the type-C behavior questionnaire for anxiety, depression, anger and optimism were respectively 43.01±7.47, 44.02±7.61, 21.56±5.26, 22.15±4.00 among the OLP patients and were 37.94±8.70, 39.58±7.35, 18.12±5.39, 24.05±3.23 among control group, with significant differences(P<0.05 for all) between the two groups. The female OLP patients had higher anxiety, depression, anger scores (43.21±6.97, 44.29±7.54, 21.64±5.09) and lower reasoning, domination, optimism scores (39.12±5.66, 16.29±3.95, 22.05±4.12) with significant differences (P<0.05 for all) compared with those of the female controls. The scores between male patients and male controls showed no significant difference. The patients with erosive lesions had higher anger score (22.94±5.26) than that of the patients without erosive lesions(20.60±5.03), with a significant difference(P<0.05). With the development of the disease, the tendency of anxiety and depression of the patients were more obvious, while optimism scores remained declining. The patients suffering more than 3 years of OLP had higher anger-toward-outside scores (17.36±3.35) than the patients suffering less than 3 years of OLP (15.19±3.99), with a significant difference (P<0.05). CONCLUSION: OLP patients showed an obvious type C behavior characteristic, especially in anxiety, depression, anger and low optimism. This research provides the C behavior characteristic of OLP for further psychological consultation or intervention during OLP treatment.

Remote Postconditioning Induced by Trauma Protects the Mouse Heart against Ischemia Reperfusion Injury. Involvement of the Neural Pathway and Molecular Mechanisms.

PURPOSE: Abdominal superficial surgical incision elicits cardioprotection against myocardial ischemia reperfusion (I/R) injury in mice. This cardioprotective phenomenon, termed remote preconditioning of trauma (RPCT), results in an 80 to 85 % reduction in cardiac infarct size. We evaluated cardioprotection and the molecular mechanisms of remote postconditioning of trauma (RPostCT) in a murine I/R injury model. METHODS: Mice were analyzed using a previously established I/R injury model. An abdominal superficial surgical incision was made 45 min after myocardial ischemia at the end of coronary occlusion, and infarct size was determined 24 h after reperfusion. RESULTS: The results indicated that a strong cardioprotective effect occurred during RPostCT (56.94 ± 2.71 % sham vs. 15.58 ± 2.16 % RPostCT; the mean area of the infarct divided by the mean area of the region at risk; p ≤ 0.05; n = 10). Furthermore, pharmacological intervention revealed neurogenic signaling involvement in the beneficial effects of RPostCT via sensory and sympathetic thoracic nerves. Pharmacological experiments in transgenic mice demonstrated that bradykinin receptors, ß-adrenergic receptors (AR), and protein kinase C were implicated in the cardioprotective effects of RPostCT. CONCLUSIONS: RPostCT significantly decreased myocardial infarction size via neurogenic transmission and various signaling pathways. This study describes a new cardiac I/R injury prevention method that might lead to the development of therapies that are more clinically relevant for myocardial I/R injury.

Identification of low potassium stress-responsive proteins in tobacco (Nicotiana tabacum) seedling roots using an iTRAQ-based analysis.

Potassium is one of the three main mineral nutrients, and is vital for leaf growth and the quality of tobacco (Nicotiana tabacum) plants. In recent years, the isobaric tags for relative and absolute quantitation (iTRAQ) method has been one of the most popular techniques for quantitative proteomic analysis. In this study, we used iTRAQ to compare protein abundances in the roots of control and low potassium-treated tobacco seedlings, and found that 108 proteins were differentially expressed between the two treatments. Of these, 34 were upregulated and 74 were downregulated, and 39 (36%) were in the chloroplasts. Kyoto Encyclopedia of Genes and Genomes pathway enrichment results suggested that metabolic pathways were the dominant pathways (10 upregulated and 14 downregulated proteins). Ten proteins involved in the pyruvate metabolism pathway increased their expression levels, and 17 upregulated proteins were enriched in the ribosomes category. To evaluate correlations between protein and gene transcript abundances, the expression patterns of 12 randomly chosen genes were examined. A quantitative real-time polymerase chain reaction revealed that the 12 genes were induced after low potassium treatment for 3, 6, 12, and 24 h. Our results demonstrate that low potassium levels affect protein profiles in tobacco roots.

[Clinical characters of pre-malignant and malignant polyp in postmenopausal women and the diagnostic value of hysteroscopy].

OBJECTIVE: To investigate the clinical characteristics of pre-malignant and malignant polyps in postmenopausal patients and to evaluate the diagnositic value of hysteroscopy in this disease. METHODS: From June 2005 to October 2014, 403 postmenopausal patients with polyps were treated in the Gynecologic Minimally Invasive Center, Beijing Obstetrics and Gynecology Hospital. There were 27 patients including 22 patients with pre-malignant and 5 patients with malignant polyps. All malignant lesions were endometrioid adenocarcinoma. The clinical data were retrospectively analyzed. Based on pathology, the diagnostic value of hysteroscopy was evaluated. RESULTS: (1) CLINICAL CHARACTERISTICS: there were 27 cases with pre-malignant and malignant polyps (group 1) and 376 cases with benign polyps (group 2). Compared the two groups, the average age was (60±8) vs (58±6) years old, the period of menopause was (9.8±8.1) vs (8.3±6.9) years. Thirteen cases (48.1%, 13/27) in group 1 and 159 cases (42.3%, 159/376) in group 2 had clinical symptoms including postmenopausal bleeding and vaginal discharge. Twelve cases (44.4%, 12/27) in group 1 and 140 cases (37.2%, 140/376) in group 2 were with hypertension. Five cases (18.5%, 5/27) in group 1 and 43 cases (11.4%, 43/376) in group 2 were with diabetes. The measures above were no significant differences (P>0.05) (2) Ultrasound features: the average thick of endometrium in group 1 and group 2 were respectively (1.3±0.7) and (0.8±0.4) cm, which had statistical significance (t=4.98, P=0.001). (3) Hysteroscopic diagnosis: the average diameters of polyp in group 1 and group 2 were respectively (2.4±1.0) and (1.6±1.0) cm, which had statistical significance (t=2.93, P=0.004). Six cases in group 1 were diagnosed by hysteroscopy including 4 cases of malignant polyp and 2 cases of pre-malignant polyp. The sensitivity, specificity, positive predictive value, negative predictive value and the accuracy were 22.2% (6/27), 100.0% (376/376), 100.0% (6/6), 94.7% (376/397) and 94.8% (382/403), respectively. CONCLUSIONS: Pre-malignant and malignant endometrial polyps are more common in the subjects with the larger diameters and the thicker endometrium. All polyps should be under complete resection by hysteroscopy and through pathology examination.

Genetic diversity analysis of fruit characteristics of hawthorn germplasm.

One hundred and six accessions of hawthorn intraspecific resources, from the National Germplasm Repository at Shenyang, were subjected to genetic diversity and principal component analysis based on evaluation data of 15 fruit traits. Results showed that the genetic diversity of hawthorn fruit traits varied. Among the 15 traits, the fruit shape variable coefficient had the most obvious evaluation, followed by fruit surface state, dot color, taste, weight of single fruit, sepal posture, peduncle form, and metula traits. These are the primary traits by which hawthorn could be classified in the future. The principal component demonstrated that these traits are the most influential factors of hawthorn fruit characteristics.

Quantitative trait locus analysis of grape weight and soluble solid content.

A grapevine hybrid population was derived from a crossing of the early-maturing female parent cultivar '87-1' and the late-maturing male parent cultivar '9-22'. A total of 149 plants were selected from the hybrid population as the mapping population, and after sequence-related amplified polymorphism and simple-sequence repeat marker analysis were conducted we constructed molecular genetic maps of the parents. The molecular linkage map of '87-1' had 19 linkage groups that contained 188 markers, with an average interval of 5.7 cM and a total distance of 1074.5 cM; the '9-22' map had 19 linkage groups that contained 175 markers, with an average interval of 7.8 cM and a total distance of 1100.2 cM. The molecular linkage map of both parents had 19 linkage groups that contained 251 markers, with an average interval of 5.0 cM and a total distance of 1264.2 cM. We used the interval mapping method to conduct a quantitative trait locus (QTL) analysis of grape weight and soluble solid content of the mapping population. Six QTLs were related to grape weight, and the average contribution to the phenotypic variance was between 11.3 and 33.0%. Seven QTLs were related to soluble solid content, and the average contribution to the phenotypic variance was between 15.7 and 55.8%.

Cladosporium cladosporioides Identified in China on Tobacco Seeds.

Tobacco (Nicotiana tabacum L.) is a leafy, annual, solanaceous plant grown commercially for its leaves. China is the biggest single tobacco market and accounts for more than 40% of the global tobacco consumption (1). Tobacco seed harvested in Guiyang, Guizhou Province, China, are commonly contaminated or infected by various fungal pathogens, which can cause abnormal seedlings with dark brown lesions and stunting of roots and decayed seeds. In 2013, five samples of 500 seeds from tobacco cv. Guiyan 4 were tested for germination on moistened paper on petri dishes. On average, 35% of the seeds from all five samples developed into abnormal seedlings or were decayed and were plated onto potato dextrose agar media and grown for 5 days at 25°C in darkness to confirm the presence of a pathogen. However, one fungus was isolated from an average of 10% of the 500 seeds sampled. It was identified morphologically as Cladosporium cladosporioides (Fresen.) de Vries based on the velvety olive-brown with almost black reverse colony color and dimensions and color of conidia and conidiophores. Conidia formed in long branched chains that readily disarticulate, single celled, elliptical to limoniform, 2 to 8 (avg. 4.3) × 2 to 3 (avg. 2.1) µm. Conidia were pale to olive brown and smooth to verruculose. Ramoconidia were 0 to 1 septate, 7 to 14 (avg. 9.2) × 2 to 4 (avg. 2.6) µm, smooth or sometimes minutely verruculose. Conidiophores were pale to olive brown, macro- and micronemateus, smooth or sometimes verruculose, and of various lengths up to 320 µm long and 2 to 5 µm wide. Primer pair ITS1 and ITS4 was employed to amplify the regions of ITS1-5.8s-ITS2 of the pathogens. Sequences of all three isolates (G3, G10, and G18) (Accession Nos. KF841547, KF841554, and KF841560) were identical to each other and to four sequences in GenBank (JX230994.1, JQ768317.1, JQ768322.1, and AB763555.1). Pathogenicity of the three isolates of C. cladosporioides was verified on tobacco seedlings of 3-week-old grown on wet filter paper in the petri dishes (9 cm in diameter). For each isolate, 20 seedlings incubated in one plate were inoculated with 0.5 ml of a suspension of 105 conidia/ml. Twenty seedlings were treated with sterile water as control treatment. After inoculation, the petri dishes were incubated at 25°C, 100 to 120 µEm-2 S-1, RH > 80%, and 16 h light per day for disease development. At 96 h after inoculation, symptoms comprising medium brown to black lesions on the roots were clearly visible on inoculated plants but not on the control plants. All seedlings inoculated died 9 days after inoculation whereas control seedlings remained symptomless. Re-isolation attempts on PDA from roots demonstrated C. cladosporioides to be present in symptomatic seedlings but not in roots of the control plants. Moreover, the characteristics of the cultured fungi were exactly the same as those originally isolated. Isolates G3, G10, and G18 (KF841547, KF841554, and KF841560) were deposited with the Tobacco Diseased Fungi, Guizhou Academy of Tobacco Sciences, Guizhou, China. Previously, C. cladosporioides has also been isolated from macadamia (Macadamia integrifolia Maiden & Betche) racemes in South Africa (4), from diseased papaya (Carica papaya L.) in Taiwan province of China (2), and from seeds of Amaranthus spp. in Poland (3). To the best of our knowledge, this is the first report of C. cladosporioides causing seed disease on tobacco in China and the disease should be considered in existing disease management practices. References: (1) British American Tobacco Annual Report, 8, 2012. (2) R. S. Chen, et al. Plant Dis. 93:426, 2009. (3) W. Pusz. Phytopathologia 54:15, 2009. (4) N. van den Berg et al. Plant Dis. 92:484, 2008.

First Report of Fusarium Wilt of Tobacco Caused by Fusarium kyushuense in China.

Tobacco (Nicotiana tabacum L.) is a leafy, annual, solanaceous plant grown commercially for its leaves. China accounts for more than 39.6% of total global tobacco production (3). In May 2012, seedlings of tobacco cv. Honghuadajinyuan in a Guiyang tobacco commercial field (Guizhou, China, 26.35° N, 106.42° E) developed symptoms of severe wilting, chlorosis, and stunting. The main stem and taproot exhibited reddish to light brown vascular discoloration; further progression of these symptoms eventually caused mortality of infected seedlings. To isolate the causal agent, necrotic tissues from the symptomatic root were placed on potato dextrose agar (PDA) and incubated at 25°C in darkness. Colonies with white to rose mycelia and red-brown colony colors developed on PDA after 5 days of incubation. Microconidia were abundant, straight or slightly curved, clavate, 0- to 3-septate, and 7.5 to 20.0 × 2.5 to 5.0 µm. Macroconidia were straight or slightly curved, slender, 3- to 5-septate, and 25.0 to 45.0 × 3.3 to 5.0 µm. Based on the observed colony attributes, growth patterns, absence of chlamydospores, micro- and macro-spore attributes (1), and PCR amplification (using primers ITS1/4) combined with translation elongation factor primers (EF1/2) (2), the fungus was identified as F. kyushuense O'Donnell & T. Aoki. Sequence of ITS1-5.8s-ITS2 region of rDNA (GenBank Accession No. JX235957) exactly matched the sequences of F. kyushuense accession AB587020.1 (100% similarity). Analysis of the elongation factor (EF-1alpha) gene of the fungus (JX658565) resulted in a 99% match for F. kyushuense accession AB674297.1. Pathogenicity of the fungus was confirmed by performing Koch's postulate as follows. Pure cultures of the fungus F. kyushuense obtained from symptomatic tissues of tobacco seedlings were grown on PDA for 6 days. Tobacco plants to be used in pathogenicity tests were germinated and grown on potting soils in a plastic container. Additional fertilization was supplied by adding 0.2 g/L of 20-20-20 (N-P-K) in the float water. When seedlings got 6-leaf stage, they were ready for pathogenicity tests. Spores harvested from these culture plates were suspended in sterile distilled water, adjusted to a concentration of 1 × 104 conidia/ml, and inoculated by irrigating 10 ml of the conidia suspension onto roots of each of the 12 tobacco seedlings with 6-leaf stage. A group of 12 seedlings of the same age treated with sterile water served as control. Inoculated seedlings were maintained at 25°C, 100 µE m-2.s-1, relative humidity >70%, and 16 h light per day, and monitored for 9 days for symptom development. Seedlings inoculated with conidia developed disease symptoms with roots with vascular discoloration of roots whereas control seedlings remained symptomless. F. kyushuense was reisolated from the symptomatic seedlings 9 days after inoculation. F. kyushuense has also been isolated from rice seeds in China (4), and from diseased wheat in Japan (1). The common tobacco Fusarium disease reported in China was caused by F. oxysporium f. sp. nicotianae. However, to the best of our knowledge, this is the first report of F. kyushuense causing wilt on tobacco in China and the disease must be considered in existing disease management practices. References: (1) T. Aoki and K. O'Donnell. Mycoscience. 39:1, 1998. (2) D. M. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (3) US Census Bureau. Foreign Trade Statistics. Washington DC, 2005. (4) Z. H. Zhao and G. Z. Lu. Mycotaxon. 102:119, 2007.

Isolation and characterization of microsatellite DNA loci from the peanut worm, Sipunculus nudus.

Sipunculus nudus, the peanut worm, is the best known species in its genus. This unsegmented subtidal marine worm is consumed in some parts of Asia and is also used as fish bait. We found 20 microsatellite DNA markers for S. nudus. The number of alleles per polymorphic locus ranged from two to seven in a sample of 39 individuals. Observed and expected heterozygosities per polymorphic locus varied from 0.103 to 1.000 and from 0.307 to 0.771, respectively. Five loci showed significant departure from Hardy-Weinberg equilibrium after sequential Bonferroni's correction. No significant linkage disequilibrium between pairs of loci was found. These microsatellite markers will provide useful tools for investigating genetic population structure, population history and conservation management of S. nudus.

Metabolomics and pathway analyses to characterize metabolic alterations in pregnant dairy cows on D 17 and D 45 after AI.

Nutrient flow to the embryo and placenta is crucial for proper development and growth during pregnancy. In this study, a metabonomic analysis was undertaken to better understand global changes in pregnant dairy cows on D 17 and D 45 after timed artificial insemination (AI). Metabolic changes in the blood plasma of pregnant dairy cows were investigated using HPLC-MS and a multivariate statistical analysis. Changes in metabolic networks were established using the MetPA method. Alterations in six metabolic pathways were found on D 17 and D 45, including variations in the level of alpha-linolenic acid metabolism, glycerophospholipid metabolism, pentose and glucuronate interconversions, glycerolipid metabolism, folate biosynthesis, and tyrosine metabolism. In addition to these pathways, 9 metabolic pathways were markedly altered on D 45. These pathways included changes in the one-carbon pool caused by folate; phenylalanine, tyrosine and tryptophan biosynthesis; thiamine metabolism; pantothenate and CoA biosynthesis; purine metabolism; inositol phosphate metabolism; amino sugar and nucleotide sugar metabolism; pentose phosphate; and the TCA pathway. The combination of metabonomics and network methods used in this study generated rich biochemical insight into possible biological modules related to early pregnancy in dairy cows.